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Dissertation/Thèse

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2024
Thèses
1
  • LEONARDO BRUNO ARAGÃO DE ARAÚJO
  • A DEFINIR

  • Leader : MARIA APARECIDA MEDEIROS MACIEL
  • MEMBRES DE LA BANQUE :
  • MARIA APARECIDA MEDEIROS MACIEL
  • DEBORAH DE MELO MAGALHÃES PADILHA
  • AUREA ECHEVARRIA AZNAR NEVES LIMA
  • DANIEL DIAS RUFINO ARCANJO
  • WESLLEY DE SOUZA PAIVA
  • Data: 29 janv. 2024


  • Afficher le Résumé
  • Colloidal systems increase the bioavailability of drugs and are largely used as an important strategy in the protection and directed delivery of bioactives compounds from synthetic or natural origin. In this sense, the therapeutic efficacy of the medicinal species Anacardium occidentale Linn. (AO) was evaluated in a nanobiotechnological study in which in vitro (antioxidant) and in vivo (pulmonary anti-inflammatory evaluation) experimental models were accessed. In that, the hydroalcoholic extract (EHA-AO) and a fraction rich in water soluble tannins (FT-AO), obtained from the stem bark of this vegetable, were loaded into a colloidal carrier SNEDDS-type system (self-nanoemulsion drug delivery system). In this context, stands out the reduction of administered therapeutic doses, reduction of toxicological risks, maintenance of the constant level of the drug in the blood (by controlled release), and prolonged efficacy of therapeutic effects, widely attributed to the colloidal systems. The choice of the carrier SNEDDS-type, consist in the fact that SNEDDS systems shows stability under water dilutions, variations of temperature and/or pH, and are effective in the controlled release (slow and prolonged), samples loaded at low concentrations, for example, can be administered orally and keep its activities. The experimental part of the study is being carried out in different stages, such as: i) bromatological and microbiological analyses of the regional raw material (stem barks of A. occidentale); ii) phytochemical procedures for obtaining the EHA-AO and FT-AO samples; iii) solubility analyses and development of the polar colloidal nanosystem O/A (oil in water) with characteristic SNEDDS, cosurfactant free; iv) application of the formulations SNEDDS-EHA-AO and SNEDDS-FT-AO in the experimental in vitro oxidation tests, as well as in an in vivo experimental model of inflammation (pulmonary emphysema in Rattus norvegicus through orotracheal papain instillation), via oral administration. The SNEDDS carrier was prepared with Tween 80 (surfactant phase), a food-based vegetable oil containing the following constituents: vitamin E, polyunsaturated, saturated fat, and a mixture of the acids oleic, linoleic and linolenic. The SNEDDS-white system (free from the samples of the vegetable A. occidentale), after physicochemical characterization (pH, surface tension, CMC, viscosity and droplet diameter), satisfactorily solubilized the samples EHA-AO and FT-AO. The antioxidant potential of the nanoproducts SNEDDS-EHA-AO (10 mg mL -1 ) and SNEDDS-FT-AO (2.5 mg mL -1 and 5.0 mg mL - 1 ) was evaluated by using in vitro experiments with significant results observed by the following tests: reducing activity equivalent to the ascorbic acid, chelating activity of the ions iron and copper, as well as the sequestration of hydroxyl and superoxide radical. The SNEDDS-FT-AO system did not show cytotoxicity to healthy cells of the RAW, Vero and L-929 strains. The in vivo pharmacological study of the SNEDDS-EHA-AO and SNEDDS-FT-AO systems, in an experimental model of lung inflammation, as well as subsequent evaluation of histopathological analyses, showed that the SNEDDS-FT-AO formulation presents the best anti- inflammatory result. These findings, together with preliminary results of the medicinal potential of Anacardium occidentale Linn., which has proven immunomodulatory, anti-inflammatory, and antiviral action, will enable the phytotherapeutic validation of the SNEDDS-EHA-AO and SNEDDS-FT-AO formulations against the Sars-CoV-2 virus, and, therefore, may contribute against the therapy against COVID-19.

2023
Thèses
1
  • TÁSSIO LESSA DO NASCIMENTO
  • Development of Bifunctional Additives Derived from Cashew Nut Liquids for Application in Diesel/Biodiesel Blends.

  • Leader : MARIA APARECIDA MEDEIROS MACIEL
  • MEMBRES DE LA BANQUE :
  • MARIA APARECIDA MEDEIROS MACIEL
  • EVERALDO SILVINO DOS SANTOS
  • ADA AMÉLIA SANDERS LOPES
  • FRANCISCO EDUARDO ARRUDA RODRIGUES
  • HÉLCIO SILVA DOS SANTOS
  • Data: 13 mars 2023


  • Afficher le Résumé
  • World concerns regarding environmental pollution and global warming have  strengthened the search for alternative solutions to the consumption of oil and its  derivatives. For this reason, biodiesel was introduced in the Brazilian energy matrix  through the National Program for Biodiesel Production and Use (PNPB), whose objective  is to implement its production and use it in a sustainable way, with a focus on social  inclusion and regional development, also aiming to reduce the dependence on imports of  fossil oil derivative, diesel oil. Biodiesel is used in the vehicular fleet through its insertion  together with diesel, which has high oxidative stability when compared to biodiesel.  However, in the diesel/biodiesel blend, the oxidation instability of biodiesel  predominates, causing long term wear and/or corrosion in the mechanical devices of  vehicles fueled with this combination. In this scenario, antioxidant agents stand out, from  which cardanol, a by-product of the cashew nut processing industry and is present in the  Cashew Nut Liquid (CNSL), is included. In this present project, the main objective was  to evaluate the potential of cardanol and some of its derivatives as fuel additives, with  antioxidant and lubricity properties. In that, eight LCC derivative additives (AD-LCC)  were synthesized, such as: 3-pentadecylphenol (compound 1), 2-tert-butyl-5- pentadecylphenol (compound 3), 2-nitro-5-pentadecylphenol (compound 4), diethyl-3- pentadecylphenyl phosphate (compound 5), 3-pentadecylphenyl diphenyl (compound 6),  tri (3-pentadecylphenyl) phosphate (compound 7), 2-tert-butyl-5-pentadecylphenyl  diethyl phosphate (compound 8) and 2- tert-butyl-5-pentadecylphenyl diphenyl  phosphate (compound 9). The AD-LCC/derivative compounds were characterized by Gas  Cromatography Coupled to Mass Spectrometer (GC/MS) and by Infrared (IR) and  Nuclear Magnetic Resonance (H1and C13 NMR) spectroscopies. The antioxidant activity  of AD-LCC/derivative products, at concentration 2000 mg/kg was evaluated by DPPH•  and Rancimat methods, in soy biodiesel synthesized in the laboratory and characterized  according to ANP Resolution No. 45 of 08/25/2014. All AD-LCC/derivative compounds  stabilized the DPPH• radical, from which compounds 1, 3, and 7 showed the highest  percentages of free radical reduction (46.5, 81.1, and 43%, respectively). In the test via  Rancimat, all AD-LCC/derivative compounds increased the induction time of soy  biodiesel, where the largest increases correspond to compounds 3 (31%), 7 (32.8%) and  9 (31%). The lubricity of the additives AD-LCC was evaluated by the Four Ball  technique, in the diesel/biodiesel mixture on the ratio 8:2, with satisfactory results,  exception for compound 4, who showed no influence on the lubricity regime in the  analyzed mixture. Comparatively, the AD-LCC/phosphorus derivatives were the most  efficient and represent a class of eco-friendly synthetic compounds, so if commercialized,  they will add biotechnological value to the fuel industries. 


2
  • JÉSSICA NAYARA GÓES DE ARAÚJO
  • Identification and functional characterization of regulatory genetic variants in Familial Hypercholesterolemia

  • Leader : VIVIAN NOGUEIRA SILBIGER
  • MEMBRES DE LA BANQUE :
  • FERNANDO MOREIRA SIMABUCO
  • LUIS ANTONIO SALAZAR NAVARRETE
  • María José Brión Martínez
  • SUSANA MARGARIDA GOMES MOREIRA
  • VIVIAN NOGUEIRA SILBIGER
  • Data: 29 mars 2023


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  • Familial hypercholesterolemia (FH) is an inherited disorder of cholesterol metabolism, characterized by prolonged exposure to high serum concentrations of low-density lipoprotein (LDL) cholesterol, which leads to a significantly increased risk of early cardiovascular morbidity and death.The main causes of FH are variants found in genes involved in the LDL clearance pathway (LDLR, APOB and PCSK9), however, causal variants are not identified in a significant portion of clinically diagnosed individuals. Therefore, variants in regions that are not usually included in diagnostic panels, could be influencing the expression of these genes, and contributing to the phenotype. The present study aimed to identify and functionally characterize genetic variants located upstream of FH-related genes. 25 volunteers clinically diagnosed with FH but without confirmatory molecular diagnosis were selected for targeted sequencing of 3 kb upstream regions (Chr19:11086362-11089361; Chr2:21044074-21047073; Chr1:55036476-55039475 - GRCh38) using the AmpliSeq for Illumina method and the Miseq Reagent Nano Kit v2. Computational analysis of sequencing results included pre-processing, mapping of the generated reads, variant calling, and visual inspection of alignments to confirm the quality of variant calling. Potentially regulatory variants were prioritized according to regulatory probability prediction algorithm, functional annotation with data from public databases, pathogenicity prediction algorithms, variant-related literature, and prediction of altered transcription factor binding. The effect of two of these potentially regulatory variants on the functioning of the promoters of their target genes was evaluated by luciferase reporter gene assays in HepG2 cells. 34 single nucleotide variants (SNV) identified in sequencing, passed the quality filters: 6 were upstream from LDLR, 15 from APOB and 13 from PCSK9. 5 variants were considered potentially regulatory and, consequently, good candidates for functional validation assays: rs36218923-T and rs538300761-T of LDLR; rs934197-T and rs9282606-A of APOB; and g.55038486A>G of PCSK9. In vitro assays revealed that rs36218923-T variant caused a significant mean reduction in luciferase activity (75.4% ± 2.9% SEM) compared to the construct containing the wild-type LDLR promoter, whereas the rs934197-T variant did not cause a significant difference in mean luciferase activity compared to the construct containing the wild-type APOB promoter.These results suggest that the rs36218923-T variant may be contributing to the decrease in LDLR gene expression and causing the FH phenotype. More studies corroborating these findings are needed to confirm the biological relevance of this variant.





     


3
  • NESTOR RODRIGUES DE OLIVEIRA NETO
  • Development and Testing of a Device for External Cardiac Pacing and Evaluation of the Efficacy of Mechanical Cardiac Stimulation
  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • BRUNO LOBAO SOARES
  • FERNANDO AUGUSTO LAVEZZO DIAS
  • FRANCISCO IROCHIMA PINHEIRO
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • HINDIAEL AERAF BELCHIOR
  • Data: 28 juil. 2023


  • Afficher le Résumé
  • The transcutaneous pacemaker has been used as therapy in cases of bradycardia with hemodynamic impairment, but it has important limitations, due to the high energy required to promote myocardial activation, causing discomfort and muscle stimulation. Thus, alternative forms of artificial cardiac stimulation have been the object of studies. Mechanical stimulation is capable of causing cardiac activation and functioning as a pacemaker, similarly to activation by an electrical stimulus. A mechanical stimulus of a certain intensity can generate a mechanical deformation in the myocardium capable of initiating myocardial excitation, resulting in ventricular contraction. This work comprises two parts: the development and testing of a prototype electropneumatic device for transthoracic cardiac pacing in humans, and the performance of an experimental study aimed to assess the efficacy the of mechanical cardiac stimulation, comparing this form of stimulation with standard electrical stimulation, regarding cardiac activation, maintenance of ventricular command and induction of ventricular arrhythmia. Hearts from 10 adult toads (402 ± 101 g) of both sexes were partially exposed outside the body and randomly submitted to mechanical or electrical stimulation. Two pairs of copper electrodes were implanted to monitor and stimulate the heart. Mechanical stimulation was performed by means of a back-and-forth movement and impact of a 2 mm diameter metal rod on the external surface of the left ventricle (aiming at the lateral wall and apex). The electrical ventricular stimulation was provided by programmed temporary pacemaker generator in demand mode connected to two pacing wire leads. After performing the threshold measurements, each form of stimulation was maintained for five minutes, with an HR of 10 bpm above the baseline HR of each animal, maintaining the same energy and HR during each phase of the experiment. The electropneumatic device tested showed a linear correlation between the mechanical energy released and the pressure of the compressed air, providing increasing energy values within the predicted range for transthoracic cardiac pacing (considering a threshold of 0.04 to 1.5 J), reaching up to about 2.8J, with the maximum pressures provided by the hospital medical air network (average pressure of 5 bar). Data of the experimental study showed that direct mechanical stimulation (similarly to electric stimulation) can be maintained continuously by five minutes in toad hearts at low heart rate without loss of capture. Nevertheless, the maximum HR during mechanical stimulation before the loss of capture was close to baseline values and significantly lower than during electric stimulation in most toads, possibly because of different types of ion channels activated in mechanical and electrical stimulation. The occurrence of ventricular arrhythmias was similar in both forms of stimulation.

4
  • MARCELO MOREIRA DE CARVALHO
  • Molecular and cytogenetics approaches on taxonomic validation of species from the Tribe Amphiprionini (Pomacentridae: Teleostei) traded in Brazil

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • DEBORAH DE MELO MAGALHÃES PADILHA
  • DÁRLIO INÁCIO ALVES TEIXEIRA
  • JONAS HENRIQUE DE SOUZA MOTTA
  • RODRIGO XAVIER SOARES
  • WAGNER FRANCO MOLINA
  • Data: 28 sept. 2023


  • Afficher le Résumé
  • Clownfish (Tribe Amphiprionini), originating from the Indo-Pacific and Red Sea, are among the taxonomic groups of greatest commercial interest in the marine ornamental industry. In Brazil, basic information about marine ornamental fish species, as well as genomic and cytogenetic characterizations, is limited, making it challenging to estimate the current diversity and origin of commercially traded species and hybrids, thus hindering their traceability and proper management. This work aimed to make a quantitative and qualitative survey of the main marine ornamental fish species traded in Brazil and contribute to the prospection of molecular and cytogenetic biodiversity of the main species of Amphiprionini in the national’s market. Samples from the most representative commercial species were obtained in specialized stores and ornamental fish husbandry in Brazil. Species and interspecific hybrids were ranked according to the volume of individuals sold, establishing their origin and taxonomic designation or popular names. The species were taxonomically validated through the sequencing of mitochondrial DNA strands (COI) and subsequent comparisons with sequences deposited in GenBank. Additionally, the seven most representative species were submitted to cytogenetic analysis using conventional methodologies (Giemsa, C-banding and Ag-RONs staining). The market analysis from the year 2015 to 2021, gathered with genetic information from clownfish species provided a biotechnological basis for taxonomic identification, fisheries management and genetics’ improvement. Data indicated this fish group is the most popular among ornamental marine species in the Brazilian market. Data generated contribute to strengthening of national marine ornamental fish farming. Additionally, the results have an impact on biological conservation, through the expansion of captive production and the consequent mitigation of extractive exploitation in natural environments.

5
  • GIOVANNA MELO MARTINS SILVA
  • Identification of Orthocoronavirinae Subfamily Sequences in Metagenomes and Metatranscriptomes and Human Coronavirus NL63 Genomic Diversity Analysis

  • Leader : RIVA DE PAULA OLIVEIRA
  • MEMBRES DE LA BANQUE :
  • RIVA DE PAULA OLIVEIRA
  • DANIEL CARLOS FERREIRA LANZA
  • THIAGO BRUCE RODRIGUES
  • JOSELIO MARIA GALVAO DE ARAUJO
  • GLORIA REGINA FRANCO
  • STELA MIRLA DA SILVA FELIPE ACÁCIO
  • Data: 29 sept. 2023


  • Afficher le Résumé
  • The development of high-throughput sequencing technologies in the last decade resulted in large repositories of raw sequencing data and metagenomic projects. Alongside, computational biology and bioinformatics became important allies and many taxonomic classification tools were created. The pandemic of COVID-19 (Coronavirus Disease 2019) has shown the importance of better investigating coronaviruses, specially human coronaviruses that impact health and public services. Among them, HCoV-NL63 is a seasonal human coronavirus that spreads worldwide, causes mild to moderate respiratory disease, and shares the same receptor for host entry as SARS-CoV and SARS-CoV-2. This study uses public databases of sequences to 1) compares two bioinformatic tools, Kaiju and Burrows-Wheeler Aligner (BWA), in identifying  from Orthocoronavirinae viral sequences in human metagenomes and 2) conduct a comprehensive phylogenetic analysis of HCoV-NL63. 1169 human samples were randomly selected (from 3670), and  taxonomic classification was performed with Kaiju (double-step analysis with different reference databases). 150 samples were found positive for CoVs using Kaiju, however, it was not possible to assembly any genomes. In addition, all 3670 samples were analyzed using BWA, with negative results for CoVs, except for one sample containing sequences from HCoV NL63. Our findings suggest that the investigated samples did not have coronaviruses, only conserved and similar sequences between organisms, which was overestimated by the Kaiju tool. On the other hand, BWA can be used to identify viromes in various sequencing products. In this study, we also used 173 publicly available spike genes sequences from HCoV-NL63 to perform phylogenetic analysis. Maximum likelihood analysis resulted in eight subgenotypes (A1, A2, A3, B1, B2, C1, C2, and C3), consolidating the lineage B division into B1 and B2. Subgenotypes B2 (20.1%), B1 (19.5%), and C3 (16.1%) were the most prevalent. Positive selection was found in S1 (V57, G96, N431) and S2 (V1177, E1206) residues. Seven non-synonymous substitutions were detected in the RBD region. Compared to other genotypes, genotype B has a remarkable number of amino acid substitutions in the S1 region. Our analysis also detected a high prevalence of recombination events within the S1 region (12–307 amino acids). Genotype A, which has the highest number of recombination events, shows a constricted period of emergence in both time and place, displaying a pattern of purifying selection. Our results highlight the importance of continuous epidemiologic and phylogenetic surveillance, which is strongly recommended in order to predict the evolution of future variants.


6
  • CAMILA FERNANDA ALMEIDA DE FARIA
  • Courtship interactions in interspecific and intergeneric crossings of species from Osphronemidae family

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • CLÓVIS COUTINHO DA MOTTA NETO
  • DANIELA CHEMIM DE MELO HOYOS
  • DEBORAH DE MELO MAGALHÃES PADILHA
  • VERUSKA DILYANNE SILVA GOMES
  • WAGNER FRANCO MOLINA
  • Data: 29 sept. 2023


  • Afficher le Résumé
  • Intra and interspecific genetic and phenotypic variability are essential aspects of genetic improvement. In this context, crossbreeding in fish with evolutionarily distinct genetic backgrounds has contributed to the production of functional products with remarkable market value. Artificial interspecific hybrids aimed at the ornamental market have enhanced morphological, physiological, or behavioral traits of economic interest. Among the most globally traded species, Betta splendens (Osphronemidae) stands out. Due to its beauty and hardiness, it drives an enormous production chain. Given its pronounced sexual dimorphism and high level of agonistic interactions among males, this species is commonly referred to as a fighting fish, primarily suggesting morphological characteristics shaped by intrasexual sexual selection. Analyses of pre- and post-zygotic reproductive isolation mechanisms involving the genera Betta, Trichogaster, Trichopodus, and Macropodus are necessary for their captive management and the development of hybridization protocols within the family. Thus, reproductive isolating mechanisms were investigated, including analyses of courtship interactions in interspecific and intergeneric crosses within the Osphronemidae family and the cytogenetic patterns of the species. The first chapter assesses the reproductive behavioral isolation of family species and its associations with the level of phylogenetic proximity. The second chapter explores the cytogenetic diversification of six commercially relevant Osphronemid species and their potential role as post-zygotic reproductive isolation mechanisms, guiding practices potentially applicable to interspecific hybridization and market enhancement within the family.

7
  • AUGUSTO MONTEIRO DE SOUZA
  • Biological studies with modified hydroxyapatite for application in bone regeneration

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • ADRIANA FERREIRA UCHOA
  • FABIANO PERES MENEZES
  • JOSE MAURO GRANJEIRO
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • SUSANA MARGARIDA GOMES MOREIRA
  • Data: 29 sept. 2023


  • Afficher le Résumé
  • Hydroxyapatite (HA) is a widely used biomaterial in clinical and pharmaceutical applications. Many studies have shown that various ionic substitutes can be incorporated into HA. This modification can produce a mineral composition more similar to natural bone tissue with promising biological characteristics for use in bone regeneration. However, these substitutions can alter the physicochemical properties of these biomaterials, making it important to assess biosafety before large-scale production and commercialization. The aim of this study was to investigate the safety and efficacy of functionalized HA-based biomaterials for bone regeneration applications. This investigation addressed important aspects, including the assessment of genotoxicity, cytotoxicity, embryotoxicity, and the ability to promote bone mineralization. To achieve this goal, this work was divided into five independent chapters, each addressing specific aspects of the analysis of biosafety and regenerative potential of these biomaterials. The first chapter of this thesis consisted of a systematic literature review with the purpose of investigating whether nanostructured compounds based on HA are free of genotoxicity. The research protocol was structured to address crucial questions related to the genotoxic profile of HA-containing biomaterials. In this study, it was found that the genotoxicity of HA-based biomaterials concerning human health is underestimated in the scientific literature. Both in vitro and in vivo models were employed to assess the genotoxicity of these biomaterials. Overall, needle-shaped HA nanoparticles tended to cause more DNA damage and chromosomal alterations compared to other forms of HA. Additionally, the results suggest that primary cells are more sensitive than conventional cell cultures in assessing the genotoxicity of these biomaterials. These findings provided a detailed overview of the current state of knowledge regarding the genotoxicity of HA-based biomaterials, highlighting the need for further research for a comprehensive understanding of the risks associated with their use in biomedical applications. In the second chapter, the focus was on evaluating the cytotoxicity and mutagenicity of HA-based biomaterials, both those functionalized with bioactive ions (CO32-, Zn2+, Sr2+) and those that were not. The main results of this analysis were that cytotoxicity tests did not reveal significant adverse effects of HA microspheres on cell culture, and no substantial changes in chromosomal levels were observed in CHO-k1 cells after exposure to different biomaterials. Moreover, there was no increase in bacterial reverse mutation rate following exposure to these biomaterials. These results provided support for the use of these biomaterials in bone regeneration applications, with confidence in their biosafety. In the third chapter, we conducted embryotoxicity tests on zebrafish to assess the safety of hydroxyapatite (HA)-based biomaterials, including HA with substituted ions. There was no significant mortality (less than 20%) or neurotoxic effects detected. Furthermore, the biomaterials did not alter oxidative stress or negatively affect development. These results support the safety and utility of these biomaterials in medical device applications and bone regeneration. In the fourth chapter, our study focused on evaluating the biosafety and bone regeneration potential of HA nanoparticles. Specifically, we investigated the preliminary biosafety of HA nanoparticles at concentrations of 25, 50, and 100 μg/mL. We used umbilical cord-derived mesenchymal stem cells as a cellular model for this evaluation. Cytotoxicity was assessed through the MTT assay, while osteogenic potential was analyzed through alkaline phosphatase activity and alizarin red staining. These assays allowed us to determine the biological safety of these biomaterials as well as their potential to promote osteogenesis. In the final chapter, we assessed the biosafety of functionalized HA nanoparticles in zebrafish embryos and larvae. We used various concentrations (50, 100, and 500 μg/mL). We monitored larval survival and hatching rates and the presence of developmental abnormalities. Additionally, we investigated the impact of these biomaterials on bone mineralization using techniques such as staining and fluorescence microscopy. Overall, the results obtained in the different chapters of this study highlight the biological safety of HA-based biomaterials, as well as their potential in osteogenesis at different concentrations. These findings have significant implications in the field of biomaterials, contributing to the advancement of bone tissue engineering and regenerative medicine applications.

8
  • NAYARA SOUSA DA SILVA
  • Clinical Characteristics Associated with COVID-19 Severity and Predisposition to Post-COVID Condition Development

  • Leader : ANDRE DUCATI LUCHESSI
  • MEMBRES DE LA BANQUE :
  • ANDRE DUCATI LUCHESSI
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • OTAVIO CABRAL MARQUES
  • PEDRO BRAGA NETO
  • RICARDO NEY OLIVEIRA COBUCCI
  • Data: 17 oct. 2023


  • Afficher le Résumé
  • Brazil has recorded approximately 37 million cases of COVID-19 as of January 2023. Besides the acute manifestations, COVID-19 might progress to a chronic disease, known as Post-COVID Condition (PCC). Longitudinal studies are essential for comprehending the long-term impacts of COVID-19. This study describes the clinical profile of a cohort of 704 patients infected with SARS- CoV-2 in 2020 and examines the health consequences and quality of life two years after infection. The first stage of the study characterized the acute phase of COVID-19 in patients infected in 2020. Critical cases accounted for 29% of the studied group, with male sex and the presence of diabetes posing higher risks for disease severity (RR=1.76; 95%CI: 1.43 – 2.17 and RR=1.60; 95%CI: 1.28 – 2.00). The use of "COVID Kit" medications—antimalarials, azithromycin, and ivermectin—was significantly higher in severe and critical severity groups (p=0.010, p<0.001, p=0.010, respectively), but no impact on patient survival was observed. A total of 105 patients died due to COVID-19 and its complications. The second stage of the study took place between May and June 2022, involving 137 individuals with the goal of investigating PCC. Persistent symptoms were reported by 56.9% of patients. Women and individuals hospitalized during the acute infection phase faced a higher risk of experiencing symptoms after COVID-19 (RR=2.07; 95%CI: 1.47 – 2.94 and RR=1.53; 95%CI: 1.07 – 2.18). Respiratory disturbances, neurological effects, and body pains were the most frequently reported manifestations. At least 20% of individuals reported some degree of difficulty in performing daily activities. Finally, countries that have encountered high rates of SARS-CoV-2 infection must be prepared to address the long-term consequences of COVID-19. In this context, our study is pivotal in providing an overview of the Brazilian situation and guiding further research as well as the formulation of targeted public policies for addressing PCC.

2022
Thèses
1
  • GILDACIO PEREIRA CHAVES FILHO
  • A DEFINIR

  • Leader : SUSANA MARGARIDA GOMES MOREIRA
  • MEMBRES DE LA BANQUE :
  • SUSANA MARGARIDA GOMES MOREIRA
  • ANA HELONEIDA DE ARAUJO MORAIS
  • CARLOS AUGUSTO GALVAO BARBOZA
  • MARIA HELENA RAPOSO FERNANDES
  • FRANCISCO MIGUEL PORTELA DA GAMA
  • Data: 19 janv. 2022


  • Afficher le Résumé
  • Health biotechnology and regenerative medicine seek alternatives to circumvent chronic or degenerative diseases, including diseases related to bone metabolism and obesity, for which conventional treatments have yet to provide satisfactory solutions. Thus, scientists have been working on the development of treatments based on biomaterials, cells and biomolecules to respond to these problems. Seaweed is a source of new molecules with potential bioactivity. Within this wide variety of molecules, sulfated polysaccharides (SP), which are highly complex and heterogeneous macromolecules, have shown interesting biological activities. Among the different types of seaweed, green seaweeds are the least studied, although several biological activities have already been described for their molecules. With regard to activities in the modeling of cell differentiation, studies with SP of green algae are particularly scarce. Thinking about that, this work evaluated the osteogenic and antiadipogenic activity of SP-containing SP samples obtained from the green marine algae Caulerpa prolifera and Caulerpa sertularioides. Additionally, as an essential part of the evaluation of the biocompatibility of new molecules, the genotoxicity of samples with biological activity was also determined. The results indicate that the SP-rich samples of C. prolifera and C. sertularioides induce differentiation of human stromal stem cells in non-cytotoxic concentrations (<10 µg / mL), promoting an increase in the alkaline phosphatase activity (ALP) and mineralization of the extracellular matrix, without genotoxicity under the conditions tested. The sample containing SP isolated from C. prolifera was also shown to inhibit adipogenic differentiation of pre-adipocytes, in non-cytotoxic concentrations (<200 µg / mL), and to reduce the accumulation of total lipids and triglycerides in the C. elegans model. Taken together, these results highlight the potential of SP in the development of alternative therapies for problems related to bone regeneration and obesity.

2
  • RICHELLY DA COSTA DANTAS OLIVEIRA
  • Analysis of human exposure to natural radiation in the municipality of Lajes Pintadas, Rio Grande do Norte, Brazilian semiarid


  • Leader : VIVIANE SOUZA DO AMARAL
  • MEMBRES DE LA BANQUE :
  • VIVIANE SOUZA DO AMARAL
  • RAQUEL CORDEIRO THEODORO
  • VANESSA BECKER
  • JULIO ALEJANDRO NAVONI
  • NATALY ALBUQUERQUE DOS SANTOS
  • Data: 28 janv. 2022


  • Afficher le Résumé
  • Brazil has one of the largest uranium reserves in the world. Usually this metal is available inside the earth's crust, however, natural and anthropic processes contribute to its redistribution throughout the environment. By treating a radioactive metal, uranium has a structure in which the soil is disintegrated into other products such as radiation and elements of varied toxicity such as radiant gas and lead. In the Brazilian semi-arid, specifically in the municipality of Lajes Pintadas / RN, there are rocky outcrops with a presence of natural ionizers that release the radiant gas and, consequently, lead to the environment. The objective of this thesis was to evaluate the carcinogenic and non - carcinogenic risk in the human population exposed to radon and lead in the municipality of Lajes Pintadas / RN / Brazil. To meet this objective, analyzes were performed with 123 individuals from the municipality, randomly selected. Results of the social questionnaire survey, analyzes the oral mucosa cells to visualize and compare possible changes caused by contact with lead and radon, as well as blood and urine, to quantify a presence of lead. In homes, people, people, people who had a purpose to quantify radium present in the air. We conclude that the diversity of information and a wide range of concentrations encountered, a great need for research in the region, with a view to assessing the real impact on human health.

3
  • MAGNALDO INÁCIO TAVARES MEDEIROS
  • Colloidal SNEDDS-type Systems Containing Croton cajucara Benth applied in experimental models of nociception, inflammation and dermal healing

  • Leader : MARIA APARECIDA MEDEIROS MACIEL
  • MEMBRES DE LA BANQUE :
  • MARIA APARECIDA MEDEIROS MACIEL
  • JOSE HERIBERTO OLIVEIRA DO NASCIMENTO
  • RICARDO LUIZ CAVALCANTI DE ALBUQUERQUE JUNIOR
  • TERESINHA GONÇALVES DA SILVA
  • VALDIR FLORENCIO DA VEIGA JUNIOR
  • Data: 22 févr. 2022


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  • Systemic chronic inflammation (SCI) and chronic wounds have been widely treated, but the adverse effects attributed to medications limit prolonged treatments. In this present study Croton cajucara Benth medicinal species was applied as an alternative therapy, by using in vivo models, aiming at to analyze its analgesia, inflammation and wound healing. In that, were used the non-volatil oil (so called fixed oil, FO) and the diterpine trans-dehydrocrotonin obtained from this herbal stem bark. After chromatographic studies, FO and t-DCTN were loaded into a colloidal self-nanoemulsion drug delivery system (SNEDDS-type) aiming at the carrier biocompatibility with FO and t-DCTN, as well as reductions of therapeutic doses and toxic risks, maintenance of the constant level of the drug in the blood (by controlled release of the SNEDDS systems), and prolonged efficacy of the systems therapeutic effects. Based on the polar colloidal O/A (oil in water) systems the formulation SNEDDS was prepared by using Tween® 80 and a vegetable oil of food use (in low concentrations), dispersed in a neutral aqueous medium (with high concentration). The target SNEDDS system was characterized by physicochemical analyses and showed droplet diameter with reduced scale (11 nm), and showed to be biocompatible with FO and t-DCTN. Since it is a system free of cosurfactant, reinforces the proposal of pharmacological application in prolonged periods, due to the reduction in the number of the carrier ingredients and also by applying minimized concentrations of the assayed bioactives from C. cajucara. The SNEDDS-derivatives were called SNEDDS-OFCC (containing 15 mg of fixed oil, rich in sesquiterpenes) and SNEDDS-DCTN (containing 5 mg of t-DCTN). These nanobioproducts were administered orally in experimental pain models through abdominal contortions and formalin tests. Treatment with SNEDDS-OFCC formulation showed 40.2% inhibition in central pain (neuropathic) and 42.8% in inhibition of peripheral inflammatory pain. Meanwhile, SNEDDS-DCTN in reducing abdominal contortions was effective in 76.7%, and in the formalin test inhibited 47.9% (neuropathic pain) and 52.6% (peripheral pain). In the models of inflammation (paw edema and peritonitis) dexamethasone (0.5 mg/kg, s.c.) was used as positive control. The inhibitor rates of leukocyte migration (by carrageenan-induced inflammation) showed significant reductions for the peritoneal cavity, with inhibitions 29.4% for SNEDDS-OFCC and 38% for SNEDDS-DCTN, in relation to the negative control group (vehicle). In the paw edema inhibition, 69.5% was observed for SNEDDS-OFCC and 89.7% for SNEDDS-DCTN. Based on these data, the SNEDDS-DCTN system was used in the experimental healing model, administered topically in Wistar rat lesions (7, 14 and 21 days). For each experimental group, the retraction index (morphometry) of the lesion and the tissue behavior (histopathological and immunohistochemical analysis) were observed. The rate of lesion retraction on the 14th day was quantitative (98.3%), compared to 43.7% for the 7th day, and also to the negative control group. The histopathological analyses sowed reduction in inflammation, with significant gains in cell proliferation and tissue maturation, compared to the control group. Since, SNEDDS-OF and SNEDDS-DCTN presented associated antinociceptive and anti-inflammatory action, as well as healing (SNEDDS-DCTN), so, they may meet a specific demand from patients who are on prolonged treatments (SCI-type) to combat pain and inflammation caused by skin lesions or chronic inflammations.

4
  • JOHERBSON DEIVID DOS SANTOS PEREIRA
  • A DEFINIR

  • Leader : MARIA APARECIDA MEDEIROS MACIEL
  • MEMBRES DE LA BANQUE :
  • AUREA ECHEVARRIA AZNAR NEVES LIMA
  • HERYKA MYRNA MAIA RAMALHO
  • JOSE HERIBERTO OLIVEIRA DO NASCIMENTO
  • MARIA APARECIDA MEDEIROS MACIEL
  • VALDIR FLORENCIO DA VEIGA JUNIOR
  • Data: 31 mars 2022


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  • Corrosion is a major problem affecting the industrial sector and carbon steel is the material most industrial applications, especially for the oil processing. The largest sources of oil are located in seas and oceans, and the metal corrosion occurs by the saline environment action. Some quinoxaline derivatives are reported in the literature as being effective agents in inhibiting corrosion. In the present study we evaluated the inhibitory capacity of a derivative oxazinoquinoxalina, N- (2-hydroxyethyl) -2,3-dihydro [1,4] -oxazinequinoxaline (OAQX), aiming carbon steel corrosion protection AISI 1018, in saline (3.5% NaCl). Studies of Resistance Polarization Linear (RPL) showed inhibition efficiencies ranging from 55% to 89% in low concentrations (6 pmm to 100.0 ppm) evaluated samples (OAQX) solubilized in DMSO. For determination of adsorption parameters, Langmuir isotherms were obtained, Temkin and Frumkin. The derivative quinoxaline the oxazinoquinoxalina consisted of the first compound of this class, reported as a corrosion inhibitor, which differs from other organic heterocyclic compounds applied as anticorrosion due to its solubilization in neutral aqueous media which promotes the study of its interactions with the metal-ligand. Due to the worsening of global environmental degradation, new practices have been adopted to control environmental impacts that consist of the development of clean technologies. In this perspective, colloidal systems based on coconut oil (OC) were evaluated as corrosion inhibitors in carbon steel AISI 1018, in saline medium (NaCl). The colloidal nano- and microemulsified OCS systems were evaluated by Linear Polarization Resistance (LPR) method. The maximum efficiencies were obtained in the range 70% to 84%. Aiming at to minimize environmental degradation, one of the systems (so called OC6, with 84% of corrosion inhibition) was used as carrier of the OAQX inhibitor, resulting in a more effective product, with maximum efficiency above 90%, representing a new anticorrosive product which consists in the development of clean technologies.

5
  • WESLLEY DE SOUZA PAIVA
  • A DEFINIR

  • Leader : HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • MEMBRES DE LA BANQUE :
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • KATIA CASTANHO SCORTECCI
  • RAQUEL CORDEIRO THEODORO
  • FERNANDA FOGAGNOLI SIMAS TOSIN
  • DIEGO DE ARAUJO SABRY
  • Data: 22 juil. 2022


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  • Oxidative stress is a cause of numerous diseases in humans, this causes a constant search for  new molecules that may have antioxidant capacity and prevent the emergence of diseases.  The objective of this work was to extract chitosan from fungi (Chit-F) and synthesize a new  molecule, conjugating it with gallic acid, which was called Chit-FGal, in order to potentiate  its antioxidant action. After collection, isolation and identification of the fungus as being of  the species Rhizopus arrhizus, it was possible to extract chitosan and after physical-chemical  tests such as high performance liquid chromatography (HPLC), Fourier transform infrared  (FTIR) and nuclear magnetic resonance (NMR) ), it was confirmed that the sample was  chitosan, with a degree of deacetylation of 86% and low molecular weight, in addition,  without the presence of proteins and phenolic compounds. After that, the synthesis of fungal  chitosan + AG (Chit-FGal) was successfully performed, confirmed by the 4x increase in the  amount of phenolic compounds in relation to Chit-F, as well as the appearance of a peak in  the region of the aromatic rings in the NMR analysis. When the 2 samples were  extracted/synthesized and characterized, the in vitro antioxidant tests were performed (Copper  Chelation, Iron Chelation, Total Antioxidant Capacity (CAT), reducing power and superoxide  radical scavenging), where the samples had activities in all the tests, however Chit-FGal  obtained results at least 50% (p <0.05) higher, demonstrating the potentiation of activities  when modifying the native molecule. When testing Chit-F and Chit-FGal in fibroblast cells  (3T3), there was no decrease in cell viability measured by MTT ((3-(4,5-Dimethylthiazolyl 2)-2,5-diphenyltetrazolium bromide) in any sample. , in addition, a protection test against  damage caused by hydrogen peroxide was performed, where Chit-FGal at a concentration of  0.05 mg/mL showed better protection. A protective effect of embryos against damage caused  by 0.06 mM hydrogen peroxide (100% live embryos) and 0.5% alcohol (93% live embryos)  was obtained. acridine orange, where both Chit-F and Chit-FGal demonstrated protective  action against cell death caused by hydrogen peroxide and alcohol. These results demonstrate  the good antioxidant potential of chitosan extracted from soil fungi of the Caatinga biome  (Chit-F), as well as the enhancement of this action by conjugating this polymer with gallic  acid (Chit-FGal), which makes this new synthesized molecule promising for use as  antioxidants with the potential to avoid a series of problems caused by oxidative stress. 

6
  • JANAINA FERREIRA ADERALDO
  • INFERTILITY AND THE GLOBAL MARKET IN ASSISTED REPRODUCTION: SITUATIONAL ANALYSIS, EVALUATION OF THE TECHNOLOGY FOR SPERM SELECTION AND SEMINAL MICROBIOTA PROSPECTION
  • Leader : DANIEL CARLOS FERREIRA LANZA
  • MEMBRES DE LA BANQUE :
  • DANIEL CARLOS FERREIRA LANZA
  • DANIELLE BARBOSA MORAIS
  • RICARDO NEY OLIVEIRA COBUCCI
  • RODRIGO VASCONCELOS DE OLIVEIRA
  • MAURICIO FRAGA VAN TILBURG
  • Data: 29 juil. 2022


  • Afficher le Résumé
  • Infertility is defined as failure to obtain a pregnancy after 12 months of regular unprotected sexual intercourse or as conditions incompatible with the meeting of gametes. Virtually all regions have experienced fertility declines in recent decades, to the point of being included in the United Nations Millennium Development Goals, which includes infertility as a global health problem. In this context, the size of the global assisted reproductive technology (ART) market was around USD 26 billion in 2019 and is expected to reach USD 45 billion by 2025. We have carried out a scoping review as a starting point for understanding the global market scenario in assisted human reproduction (AHR). To this end, we use topic modeling, an automated tool used to detect textual patterns. In this way, we identified the most covered topics in peer-reviewed texts and gray literature related to the market in AHR. We present these topics in a structured and commented way, having as main findings: a) despite the AHR market-moving billions of dollars worldwide and having expectations of growth in supply and demand in all scenarios studied, despite technological advances and insertion of technologies accessories (add-ons) over the last decades, the global success rates in RHA remain around 30% of the cycles performed; and b) the number of texts on these health technologies assessment (HTA) was insignificant, despite moving about USD 13.75 billion in 2020 and lacking significant evidence on success rates in AHR. Of the afore mentioned technologies, the use of microfluidics for sperm selection (MFSS) is in vogue. We performed the first meta-analysis to identify the potential benefits of using this device on clinical pregnancy and abortion rates in assisted reproduction treatments (ART). The results indicate a slight increase in the clinical pregnancy rate and a decrease in abortion rates about the sperm selection techniques by density gradient and swim-up; however, these results did not show statistical significance. Thus, the most robust indicators of success in ART remain maternal age, ovarian reserve, and qualitative and quantitative sperm analysis. Researchers have been dedicated to the search for immunogenic factors that contribute to the elucidation of the key factors responsible for infertility, especially in cases with no apparent cause. For this reason, we are dedicated to the first worldwide study on the application of metagenomic prospecting in seminal samples, whose choice of technique is justified by the possibility of non-directed identification of organisms. We used a pool of seminal samples provided by volunteers at the Reproductive Rights Unit at Maternity School Januário Cicco (MEJC). We found 71.3% bacterial domain, 27.6% eukaryotic domain, and 1.1% viral community. These findings expand the current view on the functional microbiota and point to future investigations as to the role of non- culturable microorganisms in the reproductive system. In addition, the Herv families identified in seminal samples deserve studies with a functional and evolutionary perspective. These data contribute to a new research front in the diagnosis of fertility- related diseases.

7
  • MAISIE MITCHELE BARBOSA OLIVEIRA
  • Evaluation the Effect of Losartan and Probiotic Lactobacillus mucosae CNPC 007 in the Experimental Model of Intestinal Mucositis Induced by 5-Fluorouracil in Mice.
  • Leader : CAROLINE ADDISON CARVALHO XAVIER DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • CAROLINE ADDISON CARVALHO XAVIER DE MEDEIROS
  • MARIA APARECIDA MEDEIROS MACIEL
  • AURIGENA ANTUNES DE ARAUJO
  • DEYSI VIVIANA TENAZOA WONG
  • JULIANA MONTANI RAIMUNDO
  • Data: 21 oct. 2022


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  • Intestinal mucositis (IM) is a side effect associated with cancer treatment resulting from chemotherapy. The cytotoxicity of IM resulting from the use of 5-fluouroracil (5-FU) promotes  damage to the mucosal epithelial structure and associated gastrointestinal symptoms that induce  the interruption of treatment by the patient. Currently, there is no efficient therapeutic protocol  that improves the symptoms of intestinal mucositis, in this sense, in order to propose  alternatives that aim at the attenuation of probable in patients affected by IM, the aim of this  study was to evaluate the effect of losartan ( LOS) and the protective effect of the probiotic (Pb)  Lactobacillus mucosae CNPC 007 on 5-FU-induced intestinal mucosite in swiss mice. 5-FU  (450 mg/kg, i.p.) was used to induce an intestinal mucositis on the 1st day. Experimental groups  included: Saline Group, 5-FU Group, LOS Group (5, 25 or 50 mg/Kg) and Probiotic Group.  Losartan was administered 30 min before 5-FU and the probiotic was administered within 14  days prior to IM induction. These were euthanized with thiopentals (90 mg/Kg, i.p.) on the 4th  day after induction and blood and intestinal segments were collected for further analysis. Blood  samples were used to count leukocytes and jejunal segments were separated for enzyme  immunoabsorbance assays for cytokine dosage (TNF-α and IL-1β), oxidative stress (MDA and  GSH) and quantitative polymerase chain reactions in time real (qPCR) for the TWEAK, Fn14  and NF-κB p65 genes. Animals with IM without major intestinal damage in the jejunum  portion, observed by shortened villi, loss of crypt architecture and intense inflammatory cell  infiltrate, evaluated by histopathological analysis with scores of 3 (variation 2-3) and  morphometry (p < 0.001 vs saline group). LOS 50 mg / Kg presented the best results for the  initial analyses, this being a dose adopted for the evaluation of the study. Leukopenia and  greater weight loss were observed in animals with MI (Group 5-FU) compared to the saline  group. Treatment of the LOS 50 and Probiotic groups improved the appearance of villi and  crypts (p < 0.01), reversing as histopathological changes (scores 1, variation 1-2) and  morphometry, leukopenia and weight loss (p < 0.05 vs 5-FU group), induced by 5-FU.  Reduction in the levels of tumor necrosis factor (TNF) -α and interleukin (IL) -1β cytokines for  the LOS 50 and Probiotic groups. LOS 50 reduces oxidative stress markers and the expression  of NF-κB genes p65, TWEAK and Fn14 compared to the 5-FU group (p <0.001), showing a  beneficial effect on induced MI in mice. In conclusion, LOS 50 mg / Kg and the probiotic L.  mucosae CNPC 007 enhance the attenuation of IM symptoms, considering that both treatments  improved the IM injury induced by 5-FU, an expression of pro-inflammatory mediators in  segments jejunals marketed. 


8
  • IASMIM SANTOS MANGABEIRA E SILVA
  • Characterization of SNPs and Microsatellite molecular markers in Penaeus vannamei and proposal of panels for population and association studies.

  • Leader : DANIEL CARLOS FERREIRA LANZA
  • MEMBRES DE LA BANQUE :
  • DANIEL CARLOS FERREIRA LANZA
  • KARINA RIBEIRO
  • RAQUEL CORDEIRO THEODORO
  • ALINE SILVA MELLO CESAR
  • JULIANA GABRIELA SILVA DE LIMA
  • Data: 20 déc. 2022


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  • Shrimp farming worldwide, and especially in Brazil, suffers profound impacts due to the lack of standardization in production processes and the occurrence of infectious diseases. In this sense, the study of the genome of the shrimp Penaeus vannamei and the development of new tools that allow the selection of animals with the vigor necessary for production is crucial for the success of the activity. Among these tools, the use of molecular markers and their association a trait of interest has shown to be a promising strategy, especially considering that vaccines and treatments are not available for shrimp farming. However, information about molecular markers for P. vannamei is still quite fragmented and many markers are not properly characterized. Thus, this work aimed to identify and characterize in detail single nucleotide polymorphism (SNP) and microsatellite (STR) markers already described in the literature for P. vannamei, and based on this characterization, propose panels of markers with potential application in population and association studies. In Chapter 1, 512 SNPs associated with genes involved in disease resistance were identified and classified in detail. It was observed that most SNPs were present in Toll-like receptor proteins 1 and 3, large and small subunits of hemocyanin and anti-lipopolysaccharide factors 1 and 2, allowing to propose their use as targets in studies of association and genetic improvement of populations of P . In chapter 2, 306 polymorphic STRs were characterized in detail, in order to identify their location in the genome. Among these, 38.8% were considered perfect, with the highest prevalence of dinucleotides (45.3%). The number of uninterrupted repetitions ranged from 3 to 62, with the majority consisting of short repetitions. Putative sequences of 135 STRs were obtained and, after functional alignment, 62 of them were associated with coding genes (mRNA). The identified proteins are mainly related to binding activity (53.5%) and catalytic activity (32.14%). Furthermore, 7 perfect STRs located in DNA coding regions were validated in vitro in a shrimp population. The average number of alleles per locus was approximately five, indicating that there may be different protein isoforms in the population. In conclusion, this work summarizes all the markers already described and characterized for P. vannamei and details panels of SNPs and microsatellites in coding regions. These markers have potential for use in population studies and, above all, for studies of direct phenotype-genotype association, increasing the study of genetic plasticity and genotype phenotype correlation in P. vannamei.

2021
Thèses
1
  • CÉSAR ORLANDO MUÑOZ CADAVID
  • Contribution of repair pathways by excision of bases and nucleotides in the development of neurodegenerative phenotypes in transgenic models of Caenorhabditis elegans

  • Leader : RIVA DE PAULA OLIVEIRA
  • MEMBRES DE LA BANQUE :
  • RIVA DE PAULA OLIVEIRA
  • JULLIANE TAMARA ARAUJO DE MELO CAMPOS
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • FERNANDA MARQUES DA CUNHA
  • CARLOS RENATO MACHADO
  • Data: 19 janv. 2021


  • Afficher le Résumé
  • Neurodegenerative diseases (NDs) are characterized by a progressive neuronal loss leading to motor or cognitive impairment. Although it is still not completely understood, NDs share many molecular mechanisms such as: oxidative stress, protein aggregation, deficiency of the ubiquitin–proteasome–autophagy system, mitochondrial dysfunction, impaired bioenergetics, dysfunction of neurotrophins and neuroinflammatory processes. The clinical observations that deficiencies in the nucleotide excision repair (NER) and base excision repair (BER) pathways cause human pathologies associated with neurological symptoms suggest that NER and BER might also play a critical role in chronic NDs. Here, we evaluated the deficiency of BER and NER pathways, especially EXO-3 and XPA-1, respectively, on the development of neurodegenerative phenotypes in Caenorhabditis elegans. EXO-3 and XPA-1 inhibition affected redox status by increasing ROS levels and up-regulating the expression of stress resistance related genes in a SKN-1 dependent manner. Similar results were found when APN1, XPC1 and CSB1 proteins were evaluated. EXO-3 and XPA-1 deficiency also activated the endoplasmic reticulum and mitochondrial unfolded protein response (UPR) and interfered in proteostasis as indicated by the reduced proteasome activity and proteasome subunits expression. Thesee alterations were associated with neurodegeneration of pan- and cholinergic-marked neurons. The neurodegeneration induced by EXO-3 and XPA-1 deficiency appears to be triggered by hyperactivation of the DNA damage sensor PARP-1 since this phenotype is rescued by PARP-1 inhibition. Inhibition of the apn-1, xpc-1 and csb-1 genes in transgenic animals for Mal de Alzheimer and Hungtinton's disease accelerated neurodegenerative phenotypes, the most marked affect were in animals treted with csb-1(RNAi).Together, these results support a model where deficiency of NER and BER pathways plays an active role generating a network of stress signals sufficiently strong to trigger neurodegeneration.

     

2
  • SINARA CYBELLE TURIBIO E SILVA NICODEMO
  • ATRAZINE REMOVAL BY Bacillus megaterium CCT 7935 AND Penicillium chrysogenum NRRL 807 FOR BIOREMEDIATION OF CONTAMINATED SOILS


  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • GORETE RIBEIRO DE MACEDO
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • EVERALDO SILVINO DOS SANTOS
  • CARLOS EDUARDO DE ARAÚJO PADILHA
  • PEDRO FERREIRA DE SOUZA FILHO
  • Data: 28 janv. 2021


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  • Atrazine is a herbicide that is still commonly used worldwide, even though it is recognized as a potent chemical-endocrine disruptor capable of causing damage to human health and the environment. Due to its high mobility and persistence, residues are still found in soils and waters. Therefore, the present Thesis aims to analyze the potential of atrazine degradation by Bacillus megaterium CCT 7935 and Penicillium chrysogenum NRRL 807, through bench experiments and in microcosms with soil, for bioremediation of contaminated areas. For data treatment, Single Factor Analysis of Variance (ANOVAOne Way) and Tukey test (p <0.05) were used. B. megaterium CCT 7935 was able to degrade atrazine in bench scale using it as a nitrogen source, especially when submitted to high initial atrazine concentrations (100 and 200 mg/L), with degradation efficiency values greater than 60%. P. chrysogenum NRRL 807 also showed great potential. Both fungusfree and immobilized forms presented degradation efficiency values above 40% when submitted to the initial concentration of 25 mg/L of atrazine, that was used as a carbon source. The removal processes were led by the citrocomo P450 enzyme complex. B. megaterium CCT 7935 and P. chrysogenum NRRL 807 were also efficient in the bioremediation of atrazine in microcosms with soil, with degradation efficiency values above 75%. Both strains used degradation metabolites as sources of nutrientes, however only B. megaterium CCT 7935 appeared to be able to make consecutive and nonsimultaneous use. Although P. chrysogenum has a small advantage in removal efficiency, it inhibited the natural microbiota, that demonstrated too, in turn, good performance in the treatment of atrazine (degradation efficiency of 76%). The results obtained by this Thesis consolidate the potential of the bioprocess developed by the microorganisms of the study in the atrazine treatment, and the bioremediation of soils is configured as an example of the several possible applications, which can also be extended to water and effluent treatment technologies.

3
  • MATÍAS FELIPE OTTO YÁÑEZ
  • SCIENTIFIC EVIDENCE AND TECHNOLOGICAL DEVELOPMENT TO TREAT SLEEP DISORDERS AND RESPIRATORY MUSCLES

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • ANTONIO JOSÉ SARMENTO DA NÓBREGA
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • ILLIA NADINNE DANTAS FLORENTINO LIMA
  • LUCIEN PERONI GUALDI
  • VERÔNICA FRANCO PARREIRA
  • Data: 27 août 2021


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  • In the literature, in the last 40 years some test modalities have been developed to assess the resistance of respiratory muscles, especially inspiratory muscles. The tests and prototypes of devices have always sought to assess two aspects that are relatively distinct, such as fatigue and resistance. There are several gaps in the literature on the best way to assess the resistance of respiratory muscles. Despite the importance of the measure, the previous studies using different models, of assessed resistance of respiratory muscles were based on devices, or prototypes and with a low number of subjects, which makes interpretation of the results very difficult. Also in recent decades, significant progress has been made in understanding sleep-disordered breathing and its consequences. There is evidence that treatment with positive airway pressure, facilitating the activity of the respiratory muscles, improves and controls the disease, but there is no solid evidence that this efficacy is repeated in special populations, such as stroke sequelae. This work aimed to develop a device within the scope of Technological Innovation and carry out two other studies on one on Health Technology Assessment (ATS) and another methodological clinical scientific study. The development of the device is in the final stages for filing a patent. As products of this PhD work we present: a product related to Health Technology Assessment: (i) a systematic review scientific article with meta - analysis (published in the Cochrane Database of Systematic Reviews) and a second clinical article related to the theme (published in. Frontiers in Physiology). The device to be patented is a device capable of evaluating the resistance of the respiratory muscles through different protocols, while the first publication is about evidencing the effects of the use of positive pressure in stroke patients, as a way to facilitate respiratory muscle activity and breathing; the second article deals with the analysis of respiratory muscle resistance techniques, through resistance tests, as a predictor of respiratory function in patients with chronic obstructive pulmonary disease.

4
  • VICTOR HUGO REZENDE DUARTE
  • THE ROLE OF THE TREML4 IN THE DEVELOPMENT OF ATHEROSCLEROTIC LESIONS

  • Leader : VIVIAN NOGUEIRA SILBIGER
  • MEMBRES DE LA BANQUE :
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • Gisele Medeiros Bastos
  • KARLA SIMONE COSTA DE SOUZA
  • LUCIANA SACILOTTO FERNANDES
  • VIVIAN NOGUEIRA SILBIGER
  • Data: 30 nov. 2021


  • Afficher le Résumé
  • Cardiovascular diseases (CVD) represent the highest percentage of morbidity and mortality in the world and its main cause is atherosclerosis. Atherosclerosis. is a multifactorial and inflammatory disease that has a long asymptomatic phase and commonly starts during childhood. Therefore, the accurate identification of atherosclerosis. is the starting point for the implementation of effective strategies for the primary prevention of CVD. TREML4 is a member of the immunoglobulin superfamily expressed in different cell types of the immune system, such as monocytes and macrophages and is related to CVD. We analyzed by qRT-PCR and genotyping the gene expression and polymorphisms (rs2803495 and rs2803496) of TREML4 in patients with Coronary Artery Disease (CAD) (n=137), Subclinical Atherosclerosis (340) and Post-Acute Infarcted Left Ventricular Dysfunction of the Myocardium (AMI) (65). Subjects carrying the minor alleles (G and C) have higher expression of TREML4 (OR 8.01, 95% CI 3.78 - 16.99, p <0.001 and OR 10.42, 95% CI 4.76 - 22 .78, p<0.001, respectively). Patients with major coronary artery lesions have greater expression of TREML4 than individuals without or with low and intermediate lesions (p<0.005). No association was observed between TREML4 expression and AS or LV dysfunction post-AMI (p>0.05). Through the in silico approach, we identified the miRNAS: miR-181a-5p, miR-200b-3p, miR-24-3p, miR-296-5p, miR-361-5p, miR-423-5p, miR-486- 3p and miR-708-5p potentially associated with TREML4. Thus, our results confirm that genetic polymorphisms influence TREML4 expression and that TREML4 expression is a potential biomarker for key steps in the progression of atherosclerosis and is not related to events prior to CAD and Acute Coronary Syndrome. Furthermore, we propose that 8 miRNAs differentially expressed in patients with CAD may be associated with inflammatory pathways potentially regulated by TREML4.

2020
Thèses
1
  • ANANÍLIA MEDEIROS GOMES DA SILVA
  • Regulation of non-coding RNAs in cardiac biological processes and Atrial Fibrillation.
  • Leader : VIVIAN NOGUEIRA SILBIGER
  • MEMBRES DE LA BANQUE :
  • VIVIAN NOGUEIRA SILBIGER
  • MARCELA ABBOTT GALVAO URURAHY
  • ANDRÉ ASSIS LOPES DO CARMO
  • NATHALIA KELLY DE ARAUJO
  • ROBERT POGUE
  • Data: 23 oct. 2020


  • Afficher le Résumé
  • Advances in molecular, cellular and genetic levels have provided increasingly new discoveries that allow the gradual understanding of key mechanisms of cardiac activity and related pathophysiologies. In this perspective, several cell-signaling pathways are activated for the process of contractility, electrical conduction and vascularization to occur effectively. However, external or endogenous factors can signal significant changes in natural biological processes, leading to cardiac pathophysiology. Among the various diseases that affect the heart, Atrial Fibrillation, AF, is highlighted as one of the main types of arrhythmia most prevalent worldwide. The diagnosis is made by ECG, however the asymptomatology of many patients and the short duration of fibrillation hamper early detection, leading to late diagnoses. Transcriptomic analyzes show that non-coding RNAs play an important modulatory activity, such as miRNAs, which regulate gene expression of mRNA and play a promising role as potential targets for the diagnosis, prognosis and treatments of AF. The activity of miRNAs, in turn, can be regulated by other non-coding RNAs, including lncRNAs and circRNAs, which perform among other functions the activity of capturing miRNAs so that gene expression can be modulated, featuring an activity called competitive endogenous RNA. The activity of miRNAs in AF and of lncRNAs and circRNAs acting on the biological processes involved in this disease were part of the main objectives of this study, in order to obtain a greater understanding of the mechanisms with which they are associated. Scientific articles based on findings about these activities in AF, including the description of the main circulating miRNAs in AF in addition to the miRNAs associated with pathological processes related to this disease, were carried out through database searches. These articles allowed a greater conception about the importance of miRNAs in the pathophysiology of this disease and other associated conditions. Additionally, in an article related to the objective of this study, the expression of circulating miRNAs in the AF event was experimentally evaluated, characterizing some potential targets differently expressed in this condition. Based on these findings, the functional activity of one of the miRNAs differently expressed in this previous study, miR-133b, in the activity of cardiac cells, was demonstrated using in vitro techniques of inhibition, suggesting that this miRNA modulates the expression of some mRNAs expressed in the heart in cardiomyocyte models and that could be regulated by different lncRNAs and circRNAs. Complementarily, through search in database, the main lncRNAs and circRNAs involved in cardiac processes associated with the pathophysiology of AF were analyzed and described, demonstrating that these molecules play a crucial role in the pathological complexity of this disease, whether with a protective or stimulating activity of progression of AF.

2
  • SAINT CLAIR GOMES BERNARDES NETO
  • Health technology assessment and technological innovation development in respiratory assessment and monitoring

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • GEORGE CARLOS DO NASCIMENTO
  • GERSON FONSECA DE SOUZA
  • VANESSA REGIANE RESQUETI FREGONEZI
  • ANTONIO JOSÉ SARMENTO DA NÓBREGA
  • VERÔNICA FRANCO PARREIRA
  • Data: 30 nov. 2020


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  • Introduction: Health Technology Assessment (HTA) is defined as a set of research methods responsible for the continuous process of systematic assessment of the outcomes generated by the use of technology provided, in order to generate scientific evidence for clinical and political decisions. The treatment of patients with respiratory failure can be done through the use of mechanical ventilation (MV), and the transition process to return to spontaneous breathing is a point that needs new technologies seeking to ensure better conditions for the assessment and prediction of positive outcomes. Current assessment methods allow analysis of momentary indices that are unable to identify the evolution of the behavior of respiratory variables. Objective: to develop technological innovation in the scope of respiratory assessment and monitoring, in addition to the development of evidence on the subject with systematic review. Results: scientific evidence on evaluation and respiratory monitoring of patients with neuromuscular disease weaning from MV was evidenced: (1) systematic review protocol entitled “Weaning from mechanical ventilation in people with neuromuscular disease: protocol for a systematic review; (2) submission of a scientific article entitled “Weaning from mechanical ventilation in people with neuromuscular diseases: systematic review”; in addition to (3) development and filing of an innovation patent entitled “Device for respiratory assessment and monitoring”. Conclusion: the device presented in the thesis, of an innovative character, allows continuous and detailed analysis of variables such as pressure, flow and volume, being able to identify early changes in the breathing pattern. Such a device will contribute to clinical applications in hospital, outpatient and home environments of patients on MV. The evidence from scientific evidence clarified the lack of an adequate protocol for assessing weaning conditions in patients with neuromuscular disease.

3
  • SUSANA BARBOSA RIBEIRO
  • Study of dexamethasone incorporated into poly (D, L-lactic-co-glycolic acid) (PLGA) polymeric nanoparticles in experimental oral mucositis

  • Leader : CAROLINE ADDISON CARVALHO XAVIER DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • CAROLINE ADDISON CARVALHO XAVIER DE MEDEIROS
  • MARIA APARECIDA MEDEIROS MACIEL
  • RIVA DE PAULA OLIVEIRA
  • JOZI GODOY FIGUEIREDO
  • RENATA FERREIRA DE CARVALHO LEITÃO
  • Data: 1 déc. 2020


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  • Oral mucositis (OM) is a frequent and limiting adverse reaction to cancer therapy, characterized by an intense inflammatory reaction and cumulative ulcer formation in the oral cavity. This study aimed to evaluate the effect of dexamethasone (DEX) incorporated into the polymeric nanoparticles (NPs) of poly (lactic-co-glycolic acid) - PGLA - in the experimental model of oral mucositis induced by 5-fluorouracil (5-FU) in Golden Sirian hamsters. The incorporation of DEX into the PLGA was performed using the solvent evaporation emulsification technique, followed by characterization with the determination of the zeta potential, polydispersity index, particle size, atomic force microscopy, stability study, determination of encapsulation efficiency, and study of in vitro release. To induce the OM model, 5-FU was administered intraperitoneally (ip) on the 1st (60mg / kg) and 2nd (40mg / kg) days of the experiment, with subsequent abrasions on the oral mucosa performed on the 4th day under anesthetic effect. The animals were distributed in the experimental groups: Normal, Mechanical Trauma, 5-FU, DEX (0.25; 0.5 or 1mg / kg) and NPs PLGA-DEX (0.1; 0.5 or 1mg / kg). On the 10th day of the experimental model, the animals were euthanized. Macroscopic, histopathological analyzes, IL-1β and TNF-α quantification by ELISA, immunohistochemistry for MMP2, COX-2, TGF-β, and NF-κB p65, immunofluorescence for markers NF-κB p65, MIF, SMAD 2/3 and p-SMAD 2/3 were performed, qRT-PCR assay to determine the gene expression of GILZ, MKP1, NF-κB p65, and AKT. Dexamethasone 1mg/kg and NPs PLGA-DEX 0.1mg/kg demonstrated anti-inflammatory effects in the experimental OM model, with a significant reduction in macroscopic and histopathological scores (*p<0.05). Treatment with DEX or NP PLGA-DEX attenuated the levels of the proinflammatory cytokines TNF-α and IL-1β (*p<0.05), reduced the immunostaining for MMP-2, COX-2, TGF-β, and NF- κB p65 (*p<0.05), suppressed protein expression of MIF, NF-κB p65, SMAD 2/3 and p-SMAD 2/3 (*p <0.05), inhibited gene expression for NF-κB p65 and AKT, and raised the mRNA to GILZ and MKP1 (* p <0.05) compared to animals with untreated OM (group 5-FU). DEX improved OM-induced by 5-FU. The use of an innovative formulation, developed with the aid of pharmaceutical technology for the incorporation of DEX into PLGA NPs, seems to optimize OM therapy while maintaining anti-inflammatory effects with reduced glucocorticoid dose.

4
  • LUCIANA DE ANDRADE MENDES
  • Scientific evidence and technological development for motor neuroprostheses

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • ANA RAQUEL RODRIGUES LINDQUIST
  • VANESSA REGIANE RESQUETI FREGONEZI
  • ANTONIO JOSÉ SARMENTO DA NÓBREGA
  • EDGARD MORYA
  • Data: 8 déc. 2020


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  • Motor neuroprostheses (MN) for foot drop is an alternative assistive device for patients with foot drop caused by upper motor neuron lesion. MN provide orthotic effect to users and also long-term benefits such as the improved gait pattern even when not using the device, as a treatment or learning effect. However, in Brazil there is low commercial availability of these devices, most devices are imported and high cost, making it practically inaccessible to its use by the Brazilian population. Considering this context, the aim of this PhD is to evaluate technological innovations related to Technological Innovation, Rehabilitation and Health Technology Assessment (HTA) applied to Stroke, one of the main pathologies of upper motor neuron lesion. We aim to highlight the efficacy, efficiency and effectiveness of motor neuroprostheses in the improvement of activities and participation of individuals with Stroke by performing systematic review. In addition, it is intended to develop an innovative prototype of MN for foot drop that is compatible with patent generation for possible industrial and commercial application in the Brazilian market. This research is under development and involves the Pneumocardiovascular Performance and Respiratory Muscles Laboratory and the Laboratory of Neuroengineering of the Federal University of Rio Grande do Norte - UFRN. The methodology consisted of two integrated axes: (i) systematic review with meta-analysis, including review protocol; (ii) development of MN device. The results comprised an article, Protocol for a Systematic Review, published in the Cochrane Database of Systematic Reviews, Volume 3, Year 2018, DOI: 10.1002 / 14651858.CD012991, entitled “Motor neuroprosthesis for promoting recovery of function after stroke”; systematic review article submitted to Cochrane's Stroke Group; and preliminary results of the neuroprosthesis device including the capture of raw signals from the thigh, leg and foot sensors. Therefore, the present study is in accordance with PhD Statute of the RENORBIO program in terms of deadlines and criteria for qualification, providing so far the production of qualified scientific evidence on the comprehensive theme that covers the whole category of motor neuroprynthesis for stroke patients, according to aspects of the ATS in partnership with the Cochrane Collaboration, as well as the development of part of an innovative technological prototype with potential for use by the Brazilian health system.

2019
Thèses
1
  • NAILTON JOSÉ BRANDÃO DE ALBUQUERQUE FILHO
  • Development and validation of carbohydrate protein bar based on powdered coconut water as a supplement for physical activity practitioners.

  • Leader : CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • MEMBRES DE LA BANQUE :
  • CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • ANTONIA DEBORA SALES
  • EDILSON MARTINS RODRIGUES NETO
  • JOSÉ FERREIRA NUNES
  • MICHEL BARBOSA DE ARAÚJO
  • Data: 15 avr. 2019


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  • Athletes and physical activity practitioners frequently consume protein supplements to achieve gains in muscle mass (hypertrophy) and strength, thereby improving performance. To do so, they make use of these supplements during and/or after exercise. In the case of practicing resistance exercises, a better post-workout recovery is also sought. Faced with this problem, the study aimed to develop and validate protein bars based on powdered coconut water (vegan and whey) as a supplement for physical activity practitioners by analyzing the efficiency and the acute effects of consumption through hormonal markers and injury enzymes muscular resistance training. Twelve physical activity practitioners aged 20-30 years, with experience in resistance exercises (bodybuilding) for at least two years, were trained three to five times a week, with a minimum duration of 30 minutes per day. The study was cross-over. Participants were divided into three groups: G1 - Control Group (commercial bar; 11 g protein); G2 - Group Treatment 1 (vegan bar based on powdered coconut water, 8.22 g protein); and G3 - Treatment Group 2 (whey bar based on powered coconut water, 10.52 g protein). One protein bar was ingested before training and other after training. The mean age of the 12 participants was 23.6 ± 3.2 years, with a body mass of 81.6 ± 14.3 kg, height of 174.5 ± 5.7 cm and BMI of 26.9 ± 4, 8 kg/m², being classified as overweight. On average, study participants performed strength training in their exercise routine at 5.2 ± 2.2 years. Regarding the hormonal serum levels, there were more relevant changes regarding insulin and free testosterone with 48 h post-training; IGF-1 and total testosterone levels remained constant at all times, regardless of the protein ingested. The LDH and CK muscle injury markers showed more marked changes only at 48 h post-training, with lower serum levels when ACP-Whey intake and similar levels between the commercial bar and the ACP-Moringa vegan bar. It is concluded that the vegan protein bar based on powered coconut water and moringa can be considered equivalent to the commercial protein bar. The ACP-Whey protein bar, based on powdered coconut water and Whey Protein Isolate, promoted a better post-workout recovery for those who practiced resisted physical activity.

2
  • THIAGO RENEE FELIPE
  • Development and validation of carbohydrate gel based on powdered coconut water for energy replacement of athletes

  • Leader : CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • MEMBRES DE LA BANQUE :
  • CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • ANTONIA DEBORA SALES
  • EDILSON MARTINS RODRIGUES NETO
  • JOSÉ FERREIRA NUNES
  • MICHEL BARBOSA DE ARAÚJO
  • Data: 15 avr. 2019


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  • The availability and metabolism of carbohydrates can influence the processes of fatigue (neurological) and peripheral (muscle). Exercise performance can be hampered by reduced carbohydrate levels, thus decreasing the muscle arousal process. Faced with this problem, the study aimed to develop and validate a carbohydrate gel based on powdered coconut water (COCOGEL) for the energetic replacement of athletes. Presenting as specific objectives: (1) To develop a gel of carbohydrates based on powdered coconut water according to the norms advocated by ANVISA; (2) Carry out the physical-chemical characterization of the carbohydrate gel based on coconut water; (3) To validate the use of coconut water-based carbohydrate gel in athletes in pre-workout, post-workout and 24 and 48 hours post-workout through: (a) the effect of subjective perception of effort on different forms of gel supplementation used; and (b) the acute post-training recovery response in the different forms of gel supplementation used. The study included 14 ultramarathon athletes, in the age group between 30 and 50 years of age, in a pre-competitive training phase. The design was cross-over. Participants were divided into two groups: G1 - Control Group (commercial gel ingestion), and G2 - Group Treatment (ingestion gel based on powdered coconut water). The training session was held at the athletic track of the Center for Integrated Child Care (CAIC-RN) and the blood samples were collected on-site, kept in appropriate containers and transported to the analysis laboratories. The evaluation of the Subjective Effort Perception (PSE) was performed through the application of the Athletes Tracking Sheet using the OMNI walking / running Scale (scores of 0-10). A confidence level of 95% was maintained in all analyzes. Considering the results obtained in the study, it is concluded that: (a) The composition of the bio-product COCOGEL fits perfectly into the legislation of "Food for Athletes" in the category of "Carbohydrate Gel", presenting quantity of metabolizable carbohydrates that meet the requirements and assisting in recovery of normal muscle function after strenuous exercise; (b) The bio-product COCOGEL, when consumed by athletes during endurance exercise (considered as strenuous), maintains glycemic levels, attenuates PSE and acts positively by accelerating post-workout muscle recovery; and (c) In addition to being a carbohydrate rich product, it may also be used in electrolyte replacement. Its 100% vegetable origin and its components of high nutritional value are presented as a market differential in the category of Food for Endurance Athletes.

3
  • RICARDO BASÍLIO DE OLIVEIRA CALAND
  • Antioxidant and neuroprotective activity of orange juice Citrus sinensis L. Osbeck (Bahia and Cara Cara) in the Caenorhabditis elegans model.

  • Leader : RIVA DE PAULA OLIVEIRA
  • MEMBRES DE LA BANQUE :
  • RENATA NASCIMENTO DE FREITAS
  • DAIANA SILVA DE ÁVILA
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • RIVA DE PAULA OLIVEIRA
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • Data: 21 juin 2019


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  • The orange juice Citrus sinensis L. Osbeck is considered an excellent food source of several bioactive compounds with properties beneficial to human health due to its high content of flavonoids, carotenoids, sugars, minerals and fibers. Some epidemiological and interventional studies have provided evidence to support the inverse correlation between orange juice consumption and the occurrence of cardiovascular disease and cancer. In this study, we used two naturally enriched oranges, originated through spontaneous mutation, called Bahia and Cara Cara. These oranges are high in vitamin C and flavonoids; however, Cara Cara orange is distinguished by being rich in carotenoids (phytoene, phytofluene, lycopene and β-carotene). Considering that oxidative damage and increased neuroinflammation are critically related to pathogenesis and neuronal loss in neurodegenerative diseases, the neuroprotective effect of some components of orange has been of specific interest in the search for effective treatments for these diseases. However, there is still a lack of in vivo studies to show the therapeutic effects of orange juice in its full and complete form, since its compounds have already been shown to have positive antioxidant and neuroprotective effects. In this context, we used the nematode Caenorhabditis elegans which is a widely established model organism for animal behavior, physiology and genetics studies. The objective of this work was to evaluate and compare the antioxidant, neuroprotective and physiological changes related to the aging of the treatment with the pasteurized orange juices Bahia and Cara Cara in the C. elegans model. To perform the experiments, the animals were synchronized in the L1 larval stage and treated at 20 ºC in petri dishes with solid nematode growth medium (NGM), with different concentrations of Bahia orange or Cara Cara, seeded with Escherichia coli OP50 or E. coli HT115. The treatment with orange juice provided positive antioxidant effects in C. elegans, such as reduction in endogenous levels of reactive oxygen species (ROS), increased survival under standard and oxidative stress conditions, and increased expression of antioxidant reporter genes g- glutamyl cysteine synthetase (gcs-1), glutathione-S-transferase (gst-4) and superoxide dismutase (sod-3) and hsp-16.2 chaperonin. Regarding the neuroprotective effects, it was observed that the orange treatment delays the profile of paralysis induced by the superexpression of the Aβ1-42 peptide in the muscle of the C. elegans model for Alzheimer's disease and decreases the polyglutamine aggregation in the C. elegans model for Huntington's disease. To investigate whether stress response signaling pathways would play an important role in juice effects, ERO quantification, oxidative stress resistance, gst-4::GFP expression, paralysis, and longevity in animals with knockdown were performed. the transcription factors SKN-1, DAF-16 and HSF-1 through interference RNA (RNAi). The results showed that the effects provided by the treatment with Bahia and Cara Cara orange juices were mainly dependent on the SKN-1 and DAF-16 transcription factors, especially SKN-1. In relation to physiological changes related to aging, the treatment increased the life span, the body movements and the rate of pharyngeal beats. We concluded that orange juice treatments provided excellent antioxidant and neuroprotective effects and were able to improve physiological parameters related to aging, which are strongly involved in the quality of life of C. elegans with emphasis on Cara Cara orange juice induced significantly better responses, possibly due to the higher carotenoid content.

4
  • MELYSSA LIMA DE MEDEIROS
  • Evaluation of copaiba oil loaded into SNEDDS system on wound healing and in vivo therapeutic processes applied on cutaneous lesions

  • Leader : MARIA APARECIDA MEDEIROS MACIEL
  • MEMBRES DE LA BANQUE :
  • CAROLINE ADDISON CARVALHO XAVIER DE MEDEIROS
  • JOSE HERIBERTO OLIVEIRA DO NASCIMENTO
  • MARIA APARECIDA MEDEIROS MACIEL
  • RICARDO LUIZ CAVALCANTI DE ALBUQUERQUE JUNIOR
  • VALDIR FLORENCIO DA VEIGA JUNIOR
  • Data: 29 juil. 2019


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  • Chronic wound management comprises an important aspect of medical practice and requires specific treatment for effective therapeutic treatment. Historical records show that copaiba oil is used preferentially to promote wound healing and treat inflammatory and infectious diseases. However, its low solubility in water limits its pharmacological use. In this sense, in the present research, self-nanoemulsion drug delivery systems (SNEDDS) developed by using low-concentration (0.5%) of copaiba oil (CO) were prepared in the presence of Tween® 80 and aqueous (neutral or saline) medium. The encapsulation of this bioactive oil in a SNEDDS-type system allows its slow and prolonged release with topical application in an experimental procedure for the healing of diabetic and non-diabetic wounds. Physical-chemical analyzes were performed to characterize the in naturaCO and also encapsulated in SNEDDS formulations. Chromatographic HRGC-MS analysis of in naturaCO allowed to identify sesquiterpenes (82.35%) and diterpenes (5.31%), in which the major sesquiterpenes are β-caryophyllene (32.84%) and germacrene D (18.78%). The major diterpene analyzed as its corresponding methyl esther derivative is copalic acid (4.44%). The CO-nanoformulations were prepared without the use of co-surfactant, which justifies its foremost importance in pharmacological applications, due to the reduction of carrier ingredients that may cause adverse reactions. Specifically, the SNEDDS contain copaiba oil and sunflower oil (1:1) as oil phase, variable percentages of surfactant (Tween class) and water on neutral or saline pH. The SNEDDS-CO systems were evaluated in therapeutic healing procedures, in which the in vivo assays for wound healing (uninfected and infected) were performed on Wistar rats (Rattus novergicus albinus). The animals were divided into groups of 5 animals (n = 5), submitted to surgery to remove cutaneous flap in the dorsal region and received daily topical treatment with SNEDDS-CO systems during the postoperative period. Histopathological and immunohistochemical analysis were performed after 7 and 14 days of treatment for both groups. Histomorphometric analysis revealed a lower number of neutrophils and lymphocytes in the treated groups (7 days), as well as increased collagenesis, proliferation of fibroblasts and increased epithelial thickening, which by comparison were more evident in the treated groups (14 days). In the control groups, the delay in tissue repair was related to the presence of keratinocytes as a function of the increased immunoreactivity for IL-1β. The highest number of cells immunocornized with FGF-2 was associated with the improvement of collagenase and the tissue repair process. In addition, an in vivocomparative approach of therapeutic procedures was performed in an experimental model of cutaneous lesions, in which the effect of low intensity laser therapy (LILT) on angiogenesis and immunoexpression of matrix metalloproteinase-2 was evaluated in the absence and/or presence of SNEDDS-CO systems.

5
  • CATHERINE TEIXEIRA DE CARVALHO
  • Valorization of “coalho”cheese whey for b- galactosidase production and applications

  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • ANA LÚCIA FIGUEIREDO PORTO
  • CRISTIANE FERNANDES DE ASSIS
  • EVERALDO SILVINO DOS SANTOS
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • GORETE RIBEIRO DE MACEDO
  • SÉRGIO DANTAS DE OLIVEIRA JÚNIOR
  • Data: 18 déc. 2019


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  • The present study aimed to produce the enzyme β-galactosidase (β-gal) using “coalho” cheese whey as biotechnological substrate by yeasts of the genus Kluyveromyces sp. and to evaluate processing strategies that enable its application in the food industry. In the first stage of this study, the co-production of β-gal and ethanol by submerged fermentation in different C:N concentrations was assessed. The yeast Kluyveromyces lactis NRRL Y-8279 presented better efficiency for co-production of β-Gal and ethanol. The maximum production of β-Gal was 21.09 ± 0.69 U/mL and ethanol 7.10 ± 0.09 g/L in 16 hours of cultivation. In the second stage, a 22 experimental design was proposed in order to develop a purification strategy to the enzyme. The parameters pH and ionic strength were evaluated in order to obtain a high purification factor without impairment in the yield. The higher levels of both parameters on study enhanced the purification factor of β-gal to 2.00, with greater influence of ionic strength on the FP response. The purified enzyme was submitted to electrophoresis that presented a band with molecular weight around 120 kDa, the enzyme of interest. In the third and final stage of the study, we analyzed the hydrolysis conditions of lactose in the “coalho” cheese whey with the immobilized form of β-Gal in 1% (w/v) sodium alginate. For the immobilization system, the immobilization efficiency reached 66% and high recovered activity was achieved. In addition, the immobilized form of the enzyme presented higher stability to pH and temperature changes. The immobilized enzyme had a slightly lower rate of lactose conversion (46%) when compared to the crude enzyme extract (53%). For the gastrointestinal simulations, around 40% of the enzymatic activity was preserved after 2 hours of exposure to simulated gastrointestinal environments. Overall, the results described here are promising for the industrial applications of β-galactosidase from K. lactis.

2018
Thèses
1
  • ALAINE DE BRITO GUERRA
  • Obtaining microbial consortia for biotechnological application in bioremediation

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • EVERALDO SILVINO DOS SANTOS
  • KATIA CASTANHO SCORTECCI
  • MARILENE HENNING VAINSTEIN
  • VALERIA MAIA DE OLIVEIRA
  • Data: 27 févr. 2018


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  • Drill cutting is characterized as a waste without adequate treatment, which has been accumulated generating an environmental liability of large proportions Thus, knowledge of the microbial community and its functions in the drilling cutting can be useful for the development of bioremediation strategies. In this context, the present work aims to identify microorganisms from drill cutting by metagenomic approach and obtain bacterial consortia and isolated bacteria that have hydrocarbon degradation activity and/or biosurfactant production, aiming at application in strategies of bioremediation. Total DNA was directly extracted from the drill cutting samples and from two consortia (enriched in different culture media) and sequenced using the Ion Torrent PGM platform. Isolated bacteria from the consortia were identified by sequencing of 16S rDNA by the Sanger method. Taxonomic analysis revealed changes at the phylum level and consequently genera among samples. While in the drilling cutting predominated phylum Proteobacteria and genus Halomonas, in the consortia greater abundance were observed for the phylum Firmicutes and genera Paenibacillus and Brevibacillus. Functional analysis using a specific database for hydrocarbon degradation and biosurfactant production (BioSurfDB), revealed that the selection not only maintained routes related to hydrocarbon degradation and biosurfactant production, but also favored some them. Growth curve in the presence of petroleum, colorimetric degradation test using the DCPIP indicator, microbial adhesion to hydrocarbons assay, emulsification tests, oil spreading Method and interfacial tension indicated capacity of hydrocarbon degradation and production of biosurfactants by the consortia and isolated. Returning to the residue to which they were obtained, the consortia selected in LB and YPD medium were able to degrade 66 and 30% of n-alkanes, respectively. So, differences in the composition of microorganisms, probably due to the enrichment stage in different media, reflected different metabolic capacities. LB medium has selected microbial community more adequate to the removal of alkanes in the drill cutting. The 16S rDNA sequences obtained from isolated strains presented high identity with representatives of the genera Brevibacillus, Micrococcus, Bacillus. Metagenomics was shown as a powerful tool in the analysis of microbial communities and the choice of bioremediation strategy. Autochthonous bioaugmentation can be an efficient alternative in hydrocarbon degradation. Method of enrichment through different medium can affect the composition of the microbial community and degradation ability.

2
  • JORGE DOS SANTOS OLIVEIRA
  • Prospecting of genes applied to hydrocarbon biodegradation and surfactant production using computational approaches

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • ADRIANA FERREIRA UCHOA
  • ANA TEREZA RIBEIRO DE VASCONCELOS
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • SUSANA VINGA
  • TIRZAH BRAZ PETTA
  • Data: 19 avr. 2018


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  • Crude oil extraction, transportation and use provoke the contamination of countless ecosystems. Therefore, bioremediation through hydrocarbon biodegradation or surfactants mobilization is an important subject, both economically and environmentally. Bioremediation research had a great boost with the recent advances in Metagenomics, as it enabled the sequencing of uncultured microorganisms providing new insights on surfactant producing and/or oil-degrading bacteria. Countless research studies are making available genomic data from unknown organisms obtained from metagenomics analysis of oil contaminated environmental samples. These new datasets are presently demanding the development of new tools and data repositories tailored for the biological analysis in a context of bioremediation data analysis. This work presents new bioinformatic tools and protocols for the domain of hydrocarbon biodegradation and surfactant production. The first tool created was BioSurfDB, http://www.biosurfdb.org/, acurated relational information system integrating data from: (i) metagenomes; (ii) organisms; (iii) biodegradation relevant genes; proteins and their metabolic pathways; (iv) bioremediation experiments results; and (v) a biosurfactant-curated list. The main goal of this repository is to gather information on the characterization of biological compounds and mechanisms involved in biosurfactant production and/or hydrocarbon biodegradation and make it available in a curated way and associated with a number of computational tools to support studies of genomic and metagenomic data. Secondly, a biogeographical distribution analysis of hydrocarbon degrading and biosurfactant producing genes was performed. It was of great importance for our reseach as it confirmed the correlation of surfactation and degradation, and therefore, the need for analysis of these events together. The results also suggested that terrestrial biomes have higher abundance of biodegradation genes, when compared to water biomes and that near-equatorial biomes have higher abundance of genes with potential for bioremediation. With more than 70 metagenomes analysed, this work was an extensive test to BioSurfDB and its publication validated the database and tools relevance. Meanwhile, the massive amount of data generated by metagenomics, presents specific computational challenges, demanding the development of new collaborative research platforms. In this thesis the Oil and Gas Virtual Research Environment (OGVRE), available at http://aleph.inesc-id.pt/vre/, was developed. The OGVRE that can support studies using metagenomic data with application to the oil and gas field through a responsive web-based working environment, providing: (ii) workflow sharing, both bioinformatics and bench; (ii) access to algorithms and tools; (iii) access to databases and datasets related with biodegradation and biosurfactants; (iv) access to processing resources; (v) access to storage resources; and (vi) access to social tools. The sharing of tailor-made computational and bench workflows is very useful in metagenomics research due to the novelty, heterogeneity and size of the data that is being generated. The final stage of this thesis corresponds to the selection of consortia enhanced for biodegradation of hydrocarbons ans surfactant production, using oil from the pre-salt layer of Brazil. The best consortium, together with a sample of pure oil were sequenced in a 16S amplicon approach and analysed for a taxonomic classification of this still largely unknown oil type, obtaining a research base for this economical important layer.

3
  • RENCIO BENTO FLORENCIO
  • RESPIRATORY KINEMATICS IN RESTRICTIVE LUNG DISEASES.

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • ANTONIO JOSÉ SARMENTO DA NÓBREGA
  • GERSON FONSECA DE SOUZA
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • JOCELINE CASSIA FEREZINI DE SA
  • VANESSA REGIANE RESQUETI FREGONEZI
  • Data: 27 juil. 2018


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  • Introduction: The respiratory system is composed of several structures from the morphofunctional point of view, but the chest wall and lungs perform great influence under the pulmonary evaluation process. Additionally, such a process depends on neurological control by the central nervous system (CNS), responsible for the control and generation of respiratory rhythm due to several neuronal groups. After the respiratory rhythm is generated, breathing is initiated by the active breathing of the inspiratory muscles, the ability to generate an inspiratory force between the external and internal environment of the chest wall (CW). What are the occurrences that limit the mobility of the patient and the pulmonary patients, can compromise the speedofthe ventilatory pattern andlung volumes, as is the case of some neurological diseases that evolve with ventilatory pattern and dependent on complementary therapies for a pulmonary reexpansion. Whatever the situations that limit the mobility of the CW or that affect the respiratory muscles directly, can compromise the efficiency of the ventilatory pattern and lung volumes, as is the case of some neurological diseases that evolve with restrictive respiratory pattern and depend on complementary therapies for pulmonary reexpansion. Aims: 1) To evaluate and compare the kinematics of the chest wall and its compartments during spontaneous breathing in subjects with restrictive disease versus healthy; 2) To evaluate and compare the kinematics of the chest wall of healthy subjects submitted to the use of three different devices for pulmonary reexpansion. Methodology: 1) Seventy-six individuals were evaluated (29 healthy, 27 with Parkinson 's disease - PD, and 20 post-stroke), through pulmonary function (spirometry), respiratory muscle strength (manovacuometry). Subsequently, the subjects were placed in a seated position to evaluate the variation of total and compartmental chest wall volumes and thoracoabdominal asynchrony by optoelectronic plethysmography (POE) for 3 minutes in quiet breathing. The subjects were also divided into groups according to the length of diagnosis to evaluate the variation of volumes and asynchrony. 2) Twelve healthy individuals of both genders with normal pulmonary function and respiratory muscle strength were evaluated by optoelectronic plethysmography (OEP) to evaluate the volume variations of the chest wall and it is compartments during the use of three different resources to promote pulmonary reexpansion: volume- oriented incentive spirometry (EI-v), positive expiratory pressure (PEP) and the combination of the two techniques mentioned above (EI-vp) were used to promote pulmonary reexpansion. The evaluations were distributed in three distinct days, one day for each resource and the first day the spirometry and manovacuometry tests were performed. Subjects were randomized (randomization.com) to the order of execution of the resources and evaluation in OEP, which occurred in 3 consecutive steps: 2 minutes of quiet breathing (QB), 2 minutes of protocol and 2 minutes of quiet breathing in recovery. The target respiration in EI-v was defined as 80% of the inspiratory capacity previously evaluated in spirometry. Breathing with the PEP adopted a free ventilatory pattern, however, a 10 cmH2O load was established for all subjects. Results: 1) Volume of the pulmonary rib cage volume (VRCp) was significantly lower in subjects with restrictive disease compared to healthy subjects (p<0.05). Post-stroke subjects with paradoxical movement presented lower volumes for the chest wall and its compartments, when compared to healthy (p <0.05), whereas the individuals in the PD group with paradoxical movement had lower values only for VRCp (p < 0.05); 2) During the use of IS-vp, a greater volume variation on chest wall (VCW) and its compartments were observed when compared to IS-v (p<0.05) and only for VCW and pulmonary rib cage (VRCp) when compared to PEP (p<0.05). The IS-vp device was able to generate a lower asynchrony between RCp versus AB (p<0.05) and RCp versus abdominal rib cage (RCa) when compared to IS-v (p<0.05). Conclusions: 1) Post-stroke individuals and with Parkinson's disease present a ventilatory pattern, with reduced chest wall volumes and presence of thoracoabdominal asynchrony, even in the absence of significant pulmonary function impairment. In addition, half of the individuals post-stroke and with PD presented paradoxical movement. 2) The combined use of EI-v with PEP was able to increase total and compartmental chest wall volumes, as well as improve compartment synchronism among compartments, appearing as an important tool in the treatment of patients with restrictive ventilatory pattern.

4
  • ESTHER FERNANDES TINÔCO VOLPE
  • TECHNOLOGICAL DEVELOPMENT OF DEVICE FOR ASSESSING ARTICULAR PARAMETERS IN INDIVIDUALS WITH CHRONIC VENOUS INSUFFICIENCY

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • ANTONIO JOSÉ SARMENTO DA NÓBREGA
  • DANIEL CARLOS FERREIRA LANZA
  • GEORGE CARLOS DO NASCIMENTO
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • JOCELINE CASSIA FEREZINI DE SA
  • Data: 23 nov. 2018


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  • The evaluation of human movement is an important parameter for therapeutic decision making and evolution reference after rehabilitation programs. Many devices have been developed, but few of them have satisfactory precision measures associated with ease of transport, handling and remote monitoring. Several chronic and degenerative diseases are characterized by compromised joint movement. Among them, Chronic Venous Insufficiency (CVI), which presents pathophysiological changes in the muscles of the lower limbs, with emphasis on the calf muscles and therefore to reduce the movement of the ankle joint. These changes determine a significant impact on the quality of life of these patients due to functional limitations caused by the disease. Thus, the development of angular assessment tools and targeted exercise programs for recovery of muscle performance and angular motion can assist health professionals in the planning and management of treatment and benefit patients with chronic venous insufficiency in reducing functional complaints and improves. Objectives: 1) To develop and execute a patent application for a device for digital measurement, therapeutic equipment and remote monitoring of joint parameters of the ankle joint and 2) assess the effects of a strengthening supervised exercise program of the calf muscle in physical performance, range of motion and myoelectric activity of the lower limbs muscles, cardiovascular hemodynamics and quality of life related to health in patients with CVI. Methods: The study is composed of two types of integrated research modalities: 1) Technological development and patent deposit of a device for evaluation of joint parameters and 2) randomized controlled clinical trial with blinded evaluator. Results: 1) creation of a device for evaluating human movement applied to the health area, called: "Digital Meter Device, Therapeutic Equipment and Remote Monitoring of Articular Parameters" with patent filing at INPI (BR 10 2018 016524 0); 2) Development of a strengthening exercises protocol of the calf muscles, directed to individuals with CVI and 3) Randomized clinical trial. The results of the randomized clinical trial showed that the patients in the Intervention Group - IG showed improvement in functional performance (p <0.0001) as well as in the Control Group - CG (p = 0.02). There was a decrease in the myoelectric activity in the gastrocnemius muscle during the end of the heel-rise teste (p = 0.01) at 25% and 50% moments in the IG patients, as well as a decrease in dyspnea after the end of the heel-rise test (p= 0.04). Quality of life was positively impacted in the domains: perception of improvement of symptoms in the lower limbs (V3, p = 0.01), performance in daily life activities (V5, p = 0.03), pain (V7, p= 0.04) and psychological impact of disease on the person's life (V8, p = 0.04) in IG patients. Conclusion: The present study develop a prototype, portable and precise, for angular evaluation, therapeutic training and remote monitoring to be used in patients with CVI. Furthermore, the results of the randomized clinical trial demonstrated an improvement in physical performance, myoelectric activity of the lower limbs and quality of life in treated individuals.

5
  • FRANCISCO JOSÉ CARVALHO MOREIRA
  • Prospecting extracts of Tephrosia toxicaria Pers. for control of root-knot nematodes and development of bionanformulations derived from extracts for application as nematicides in agriculture.

  • Leader : ADRIANA FERREIRA UCHOA
  • MEMBRES DE LA BANQUE :
  • ADRIANA FERREIRA UCHOA
  • DANIEL CARLOS FERREIRA LANZA
  • GORETE RIBEIRO DE MACEDO
  • LEONARDO LIMA PEPINO DE MACEDO
  • LUDOVICO MIGLIOLO
  • MARIA APARECIDA MEDEIROS MACIEL
  • Data: 30 nov. 2018


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  • Among the phytonematoids, the genus Meloidogyne stands out, due to its genetic plasticity, versatility and adaptability, which places them as the main group causing damage to agriculture. Several compounds of plant origin have nematicidal activity, are profitable and environmentally safe, and can be used in organic farming systems, however, many of these products degrade easily, reducing persistence, making control difficult. In this sense, the nanoencapsulation of botanical pesticides presents as a formidable technology, being able to transform the physical state of the substances for better manipulation, allowing the optimization of formulations with controlled release, thus avoiding their degradation, due to adverse conditions. The objective of this work was to evaluate the in vivo nematicidal activity of the aqueous and hydroalcoholic extracts of stems, leaves and roots of Tephrosia toxicaria in the control of root-knot nematodes (Meloidogyne enterolobii and M. javanica) in 'Pati' tomato their environmental evaluation through cytotoxicity in 3T3 cells and soil ecotoxicity in Caenorhabditis elegans. Then another set of assays to evaluate the in vitro effect of the aqueous and hydroalcoholic extracts free and conveyed in SNEDDS colloidal system of the roots of T. toxicaria on root-knot nematodes (M. enterolobii and M. javanica) and their environmental evaluation, with aquatic ecotoxicity in microcrustaceans (Ceriodaphinia dubia) and phytotoxicity in lettuce (Lactuca sativa). The results demonstrate that aqueous extracts of leaves, stems and roots of T. toxicaria were efficient in the control of M. enterolobii and M. javanica, and the leaves and roots were more promising, from the concentration of 0.8 %, revealing the potential of these for the development of alternatives for the control of gnats nematodes with low cost and low environmental risk. As for the second assay, it was observed for the free aqueous extract, inhibited hatching and caused mortality (100%) of the J2 from 0.2%; while free hydroalcoholic extract inhibited hatching from 0.1% and total mortality from 0.2%, with similar behavior for both nematodes. As the hydroalcoholic extract, which gave greater inhibition of hatching and mortality of juveniles. With the introduction of the extracts in colloidal SNEDDS (Self-nanoemulsifying drug delivery systems) containing aqueous and hydroalcoholic extracts of roots of T. toxicaria were effective, in vitro, in the inhibition of hatching and on the mortality of J2, inhibition complete of hatching from 0.10 and 0.025%, respectively; and total mortality from 0.050% in the two types of extracts conveyed; it was also verified that the free extracts caused mortality in only the two highest concentrations (1.6 and 3.2%), whereas the extracts showed no aquatic toxicity to the microcrustacean (C. dubia); as for phytotoxicide on germination of lettuce (L. sativa) seeds 'Summer Nanny' there was no harmful effect. Considering these results, we can conclude that the polar extracts of T. toxicaria present potential for application as a botanical nematicide. The SNEDDS system presented efficiency as a carrier system of the biomolecules of aqueous and hydroalcoholic extracts of T. toxicaria, maintaining the nematicidal effect of the active principles, demonstrating potential as a new input for the management of gall nematodes. The data set indicate the extracts of T. toxicaria as an alternative to the control of gall nematodes, with significant effect at low concentrations and with reasonable environmental safety. The SNEDDS, system carrier of the polar extracts demonstrated potential as a new input for the management of gall nematodes, which resulted in a patent deposit with the National Institute of Industrial Property (INPI), therefore, a formulation that could be used commercially in the agriculture.

2017
Thèses
1
  • PRISCILLA MOURA ROLIM
  • A DEFINIR

  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • GORETE RIBEIRO DE MACEDO
  • RENATA ALEXANDRA MOREIRA DAS NEVES
  • ANA LÚCIA FIGUEIREDO PORTO
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • SAMARA ALVACHIAN CARDOSO ANDRADE
  • Data: 10 févr. 2017


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  • The use of food processing residues is a strategy to contribute to reducing waste and adding value to new products. Melon is a fruit consumed worldwide and its high consumption accompanies large amounts of residues, such as peel and seeds. The objective of this thesis was to add value to melon peel and seeds (Cucumis melo var. reticulatus), evidencing its nutritional properties, antioxidant capacity, anti-tumor effect, prebiotic potential and its capacity for enzyme production. Flours of melon peel and seed were elaborated using experimental design 22 to evaluate the factors of drying time and temperature, and the responses moisture, water activity and flour yield. Flour characterization was carried out using centesimal composition (moisture, ash, proteins, lipids, dietary fiber), fibrous fractions of cellulose, hemicellulose, lignin and pectin. Flours were evaluated for their in vitro prebiotic properties using Bifidobacterium lactis in submerged fermentation and bile salt tolerance. The potential of the flours in the production of cellulases in semi-solid fermentation using Aspergillus oryzae fungus was also evaluated. For the accomplishment of the analyzes of antioxidant and antiproliferative power, aqueous extracts, hydrometanolic and hydroethanolic of the flours were prepared, characterized as monosaccharide composition, analysis of total sugars and proteins. For antioxidant evaluation, the total phenolic compounds, total flavonoids, total carotenoids and tannins were determined. Phytochemical evaluation of the extracts was carried out by Thin Layer Chromatography. The antioxidant capacity evaluation was verified by the following in vitro tests: total antioxidant capacity, reducing power, iron and copper metal chelation, sequestration of DPPH, hydroxyl and superoxide radicals, and evaluation of the oxygenated radicals absorption capacity. The antiproliferative activity was evaluated using the tumor cell lines (SiHa, HeLa, 786-0 and HT29) by colorimetric test of cell viability evaluation. The results for the drying experimental design showed that the temperature factor significantly influenced the yield response of the flours, being the test of 80 ºC for 24 h which presented better yield. Data showed that the melon residues had important nutritional characteristics for human consumption, with emphasis on dietary fiber content (51.75% for seed, 40.57% for peel), proteins (22% for seed; 18% for peel) and lipids (24% for seed). The residues can be characterized as cellulosic material, with values of 35% cellulose for melon seed and 19% for peel. Satisfactory percentages of pectin were detected, being 28.96% for seed and 32.65% for melon peel. As for the prebiotic potential of the flour, it was observed that the fructo-oligosaccharide standard showed a growth of bifidobacteria of 12 LogUFCmL-1, after 12 hours of fermentation, and the melon seed flour promoted bifidobacterial growth reaching 9.9 LogUFCmL- 1, after 8 hours of culture, indicating prebiotic properties, with tolerance to the action of bile salts for up to 8 hours of fermentation. The melon peel did not provide growth of bifidobacteria. Melon peel and seed proved to be a good substrate for enzyme production. The best CMCase activity (1.045 U / g) was verified on six days of fermentation and FPase (0.190 U / g) after 96 h of fermentation with medium containing the peel and seed flours. In the evaluation of the extracts obtained from the melon residues, phytochemical analysis indicated the presence of compounds characteristic of phenolics, terpenes and saponins. Total phenolic compounds in aqueous, hydrometanolic, and hydroethanolic extracts of melon residue were quantified and expressed as gallic equivalents, highlighting peel extracts (1.016 mg 100g-1). Total carotenoids, total flavonoids and tannins were also detected in extracts of melon residues, with emphasis on 96 μg of total carotenoids per gram of melon peel, 491 μg of catechin equivalents and 309 μg of catechin equivalents in 100 g extract dried of melon peel. The tests to evaluate the antioxidant activity revealed that all extracts presented antioxidant capacity. In the test of total antioxidant capacity (expressed as equivalent in ascorbic acid), hydroethanolic and methanolic extracts for peel and seed presented values of 89.74 and 83 mg / g, respectively; 50.56% and methanolic 67.7% for melon peel. Reducing power and chelating abilities of copper and iron metals presented relevant data for both melon peel and seed, demonstrating an excellent antioxidant potential. The aqueous extract of the peel showed the ability of in vitro of copper ions chelation (84%) and iron ions chelation (61%). Scavenging activity of the DPPH radical was significant only for extracts of melon peel (29.5%).The cell viability test allowed us to observe that the extracts were able to extract the organic extracts, to significantly decrease the viability of the HeLa, SiHa and 786-0 cancer cells. Aqueous seed extracts at the 1.0 mg / mL concentration inhibited the proliferation of SiHa cells by 80% and the peel hydroethanolic extract (0.25 mg / mL) inhibited by 80% the viability of HeLa cells. For human renal tumor cells (786-0) all extracts at the concentration of 1.0 mg / ml inhibited cell proliferation by more than 50%. The extracts evaluated did not present significant inhibition activity of HT29 cells. All melon peel and seed extracts weakly inhibited the proliferation of normal 3t3 fibroblasts (25% inhibition). It is concluded that melon peels and seeds have antioxidant potential in vitro and antiproliferative effects in tumor cells. Flours of melon peel and seed may be substrates for the production of cellulolytic enzymes, in addition the seed flour showed in vitro prebiotic indication.

2
  • ANDRE LEANDRO SILVA
  • Study of amphotericin B molecular aggregation into different carrier systems

  • Leader : ERYVALDO SOCRATES TABOSA DO EGITO
  • MEMBRES DE LA BANQUE :
  • ERYVALDO SOCRATES TABOSA DO EGITO
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • MATHEUS DE FREITAS FERNANDES PEDROSA
  • BOLIVAR PONCIANO GOULART DE LIMA DAMASCENO
  • FRANCELINE REYNAUD
  • Data: 20 avr. 2017


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  • The amphotericin B (AmB) is a drug of peculiar physicochemical features: being amphiphilic and amphoteric. These characteristics turn difficult the drug load into therapeutic systems. AmB is currently available in the market as micelles, liposomes and lipid complex for injection. The literature show that there is an intimate correlation between the AmB bound to the carrier and its biological response. However, there is a deficiency concerning the physicochemical characterization of the available AmB-containing products. Therefore, the aim of this work was to characterize AmB-containing carriers seeking a prediction to its biological response. The AmB-containing micellar system was the first product available for clinical use. The patent of this product has already expired some years ago. In this work we have characterized the original system and two other similar micellar products. In addition, we studied the stability increase of heated systems, by the formation of AmB “super-aggregates”. AmBisome®, an AmB-containing liposomal system, was also characterized and, for the first time, tested for the possibility of super-aggregates formation. The AmB incorporation into nano and microemulsion systems was presented and the physicochemical characteristics evaluated, focusing mainly on applications for the treatment of fungal ocular diseases and also for visceral leishmaniasis. The main techniques used for characterization were electronic spectroscopy, circular dichroism and dynamic light scattering. The isothermal titration calorimetry (ITC) was used as an attempt to measuring the super-aggregates energy formation. Besides, an AmB soluble derivative was developed and characterized by atomic mass spectroscopy, infra-red, UV-Vis and circular dichroism. Then, this AmB-derivative was loaded into a microemulsion as a vehiculation strategy. The overall results show that the AmB-containing systems presented different molecular aggregation states that depends on the carrier, the way the drug is incorporated and also on the diluent. According to the literature, the aggregation state is associated with both, drug efficiency and toxicity. In nanoemulsion systems, the drug is found aggregated and multi-aggregated. In microemulsions, AmB is loaded as monomers. The heated micellar systems form AmB super-aggregates while the liposomal system is unable to form such molecular structure. Moreover, the AmB soluble derivative presented distinct features when compared to the original molecule. However, once incorporated into the microemulsion, the aggregation state is similar to that of the original AmB molecule, as supported by UV-Vis and circular dichroism results. It can be concluded that the AmB aggregation state varies according to the kind of carrier, the drug concentration and also the way of drug incorporation, even into one same carrier. Finally, the soluble derivative opens the possibility for drug carrying into aqueous vehicles for the treatment of many diseases by different administration routes.

3
  • RODRIGO XAVIER SOARES
  • Prospecting of cytogenetic markers on large marine pelagic fish

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • ALLYSON SANTOS DE SOUZA
  • GIDEÃO WAGNER WERNECK FELIX DA COSTA
  • PAULO AUGUSTO DE LIMA FILHO
  • PAULO SERGIO MARINHO LUCIO
  • WAGNER FRANCO MOLINA
  • Data: 12 mai 2017


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  • The big pelagic fish represents one of the most lucrative subjects of the industrial fisheries in Brazil. The State of Rio Grande do Norte attracts special attention because its geographic position constituting nowadays the region in which the commercial oceanic fisheries tends to enlarge with new policies of governmental incentive. Different from other World’s producer regions that are researching the main exploited species, in Brazil the studies that objectify the population delimitation and to list data for the conservation of the species are scarce. Among the pelagic groups is common to observe a predominance of a minor diversity in relation to the abundance, this is a characteristic in two of the most important families of this oceanic region, Sphyraenidae (barracudas), Carangidae (jacks and pompanos) and Coryphaenidae (dolphinfish). Its representatives are valued and intensely exploited by the fisheries industry. Data about these species are surrounded of logistics difficulties for its size, capture way and habitat which occupies. The first cytogenetical informations for some species of this family were obtained only recently. Here is presented a cytogenetical study with 9 pelagic fish species from the Atlantic, of the families Sphyraenidae, Carangidae, Corypahenidae, Istiophoridae and Megalopidae and 1 Indo-Pacific Carangidae. The results revealed numerical divergences and in the karyotipical macrostructure contrasting with the pattern considered basal for the Teleosts, besides the existence of masculine heterogamety, through a system of multiple sexual chromosomes of the kind X1X1X2X2/X1X2Y. Using techniques such as AgNORs, MM/DAPI it was possible to identify single ribosomal sites and showed cytotaxonomic markers for all. It was also applied more resolutive cytogenetic techniques based on multigenic sequences mapping through FISH (Fluorescence in situ Hybridization) enabled for the first time for some of these families to infer about the active karyoevolutive processes and evolutive aspects of the investigated species. Through the developed techniques it will be also possible to expand the knowledge about this important marine resource,subsidizing future policies of the management of stocks, as well as future support for the technological development of marine aquaculture.

4
  • MARCELINO GEVILBERGUE VIANA
  • Isolation of endophytic microorganisms of Lippia gracilis Schauer (Verbenaceae),
     evaluation of plant essential oil and endophytic metabolites on microbiologically
     induced corrosion (CMI) in AISI 1020 carbon steel."
  • Leader : EVERALDO SILVINO DOS SANTOS
  • MEMBRES DE LA BANQUE :
  • EVERALDO SILVINO DOS SANTOS
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • IVANILDO JOSÉ DA SILVA JUNIOR
  • JADSON DIOGO PEREIRA BEZERRA
  • MATHEUS DE FREITAS FERNANDES PEDROSA
  • Data: 31 août 2017


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  • The corrosion microbiologically induced (CMI) reaches all the sectors of production of the industry in Petroleum oil and Gas and demands expressive expensive and investments for prevention and control. The goal of this thesis was to evaluate the biocide and antibiofouling activities of secondary metabolites obtained from endophytic microorganisms of Lippia gracilis Schauer as well of the essential oil (pure and in microemulsified system) from this species on microbial biofilms in 1020 AISI carbono steel. The experiments for biocide activity used 1020 AISI carbon steel plate arranged in a dynamic system with forced circulation for obtaining of the total biofilms from water produced by oil. The assessment for biocide of the metabolites from endophytic of L. gracilis Schauer resulted of the quantification of these species of bacteria in the quantification techniques by absorbance in more probable numbers (MPN), before and after the treatments, had been identified by gas chromatography coupled the spectrophotometry in mass (CG-MS) and magnetic resonance imaging (MRI) just those who demonstrated activities. The treatments consisted in evaluating the concentrations 0, 100, 150 and 200 μg/L in each metabolites on biofilms in times contact to zero, two, three and four weeks, with three repetition by treatments. The antibiofouling action was made in the contact of the metabolites in the highest concentration (200 μg/L) with the bioplates before being inserted into the dynamic system. The antimicrobial action of the essential oil L. gracilis Schauer in the pure form and in a microemulsified system was evaluated by the quantification bacteria reducer of sulphate (BRS), oxidant of iron (BOI), fungi, aerobic bacteria and total anaerobes by the techniques of dilution, platingpourplate, spectrophotometry in mass (MS) and micrographs analysis made before and after the treatments. The anticorrosive activity of all bioproducts was evaluated by electrochemical impedance and polarization, gravimetric examinations of mass loss and quantification of pites, as well in the obtaining of micrographs of the carbon steel`s surface. Samples of L. gracilis Schauer were collected in the city of Mossoró - RN and were isolated fungi and bacteria from the roots and leaves. A new type of fungus was discovered and identified as Achaetomium lippae. Three metabolites were obtained: two fungi (LGS-Lf1 and LGSLf2, being "LGS" for L. gracilis Schauer, "L": leaf, ―f‖: fungus and "R" for root) and one of bacteria (LGS-Rb1, being "LGS" for L. gracilis Schauer, "R": root and "b": bacterium). The results showed that there was a difference in the biocide activity and antibiofouling between the metabolites from endophytic of L. gracilis Schauer assessed, being just the LGS-Lf1 being efficient over all the microorganisms tested, avoiding the formation of biofouling on the surface of the material and reducing the rate of microbial growth at zero after 96 hours of contact at the highest concentration evaluated. The electrochemical and gravimetric tests displayed that for all the metabolites occured change in the anticorrosive activity. The L. gracilis Schauer essential oil evaluated in the pure form inhibited significally the growth in vito of C. bifermentans and of fungi in biofilm, reducing their growth rate to zero as from 20 μg.L-1 . The electrochemical and gravimetric tests showed that the concentration of 60 μg.L-1 of the essential oil was efficient in the reduction of corrosion. In the tests, using the microemulsified system, the essential oil inhibited the growth of BRS, BOF and fungi 16 days later in contact and had antibiofouling activity after 96 hours in contact with the AISI 1020 bioplates of carbon steel. Therefore, the metabolites from endophytic of L. gracilis Schauer as well its essential oil are shown as new alternatives of natural biocides efficient in the control of the corrosion microbiologically induced.

5
  • FRANCINALDO LEITE DA SILVA
  • Production of lignocellulolytic enzymes and bioethanol with pretreated carnauba (Copernicia prunifera) straw residues 

  • Leader : EVERALDO SILVINO DOS SANTOS
  • MEMBRES DE LA BANQUE :
  • EVERALDO SILVINO DOS SANTOS
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • MARIA VALDEREZ PONTE ROCHA
  • NATHALIA KELLY DE ARAUJO
  • TELMA TEIXEIRA FRANCO
  • Data: 19 déc. 2017


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  • Native to Brazil, Carnauba (Copernicia prunifera) has been used for several purposes, including the wax production from its leaves, in the process that generates a considerable amount of residue.  This residue is characterized as a fiber rich in cellulose and therefore with potential latent for use as a source of carbon for the production of cellulolytic enzymes and bioethanol. However, the chemical structure of this material presents cellulose bound to structurally complex components, such as hemicellulose and lignin, which hinders the production of cellulases by filamentous fungi, as well as its enzymatic hydrolysis, being essential to use of a pre-treatment for the viability of these processes. The present study evaluated the effect of different pre-treatments on carnauba straw for the production of lignocellulolytic enzymes and for the enzymatic hydrolysis with a view to the production of cellulosic ethanol through the concepts of biorefinery and micro-distillery. In the first stage, carnauba straw residue was submitted to hydrothermal (HT), alkaline (AL), alkaline acid (AA) and alkaline hydrogen peroxide (A-HP) pre-treatments. The pretreated and untreated residues were chemically characterized according to the National Renewable Energy Laboratory (NREL) protocol and, physically, by Scanning Electron Microscopy (MEV), X-Ray Diffraction (XRD) and Spectroscopy of Infrared by Fourier Transform (FTIR). A part of each residue was used to produce enzymes by means of Solid State Fermentation (FES), using the fungus Trichoderma reesei CCT-2768. The FPAse, CMCase, β-glycosidase and xylanase activities of the extracts were estimated and the production was subsequently optimized. The other part of the residues was subjected to Saccharification and Simultaneous Fermentation (SSF) using commercial enzymes and Saccharomyces cerevisiae UFLA CA11, Saccharomyces cerevisiae CAT-1 and Kluyveromyces marxianus ATCC-36907. The results of the pretreatments AL, AA and A-HP stood out in terms of the removal of lignin, according to the chemical and physical analysis of the residues. The studies showed that pretreatment of carnauba straw with A-HP has a higher capacity to induce the production of lignocellulolytic enzymes when compared to other residues, such as coconut, cashew apple and sugar cane, pretreated by the same method. The optimization of the production of lignocellulolytic enzymes allowed the production of an enzymatic extract with FPase activity of 2.4 U/g and xylanases of 172 U/g. The application of the enzymatic extract in the hydrolysis of pretreated sugarcane bagasse showed efficiency of 86.96%. The use of AL pretreated carnauba residue in enzymatic hydrolysis, with commercial enzymes, showed a higher conversion of sugars (64.43%) and, when submitted to SSF, produced 7.53 g/L of ethanol, using Kluyveromyces marxianus ATCC-36907 cultured at 45 °C. The results showed, therefore, the biotechnological potential of the carnauba residue for the production of cellulolytic enzymes and the production of bioethanol by means of biorefinery and micro distillery.

2016
Thèses
1
  • GISLAYNE CHRISTIANNE XAVIER PEIXOTO
  • Aplicação de Biotécnicas para Monitoramento e Controle do Ciclo Estral de Espécies Silvestres do Bioma Caatinga. 

  • Leader : ALEXANDRE RODRIGUES SILVA
  • MEMBRES DE LA BANQUE :
  • ALEXANDRE RODRIGUES SILVA
  • ALEXSANDRA FERNANDES PEREIRA
  • ANA LIZA PAZ SOUZA
  • LUCIA DANIEL MACHADO DA SILVA
  • NEI MOREIRA
  • Data: 4 mars 2016


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  • O objetivo da presente tese foi aplicar biotécnicas no monitoramento e controle do ciclo estral de espécie silvestres do bioma caatinga, como ferramenta para aperfeiçoamento do manejo em cativeiro, bem como conservação ou multiplicação dessas espécies. A tese foi dividida em 3 experimentos. No primeiro, foi realizado o monitoramento do ciclo estral de tatus-peba (Euphractus sexcinctus) por diferentes métodos de monitoramento, na qual foi observado ciclo estral com duração de 23,5 ± 3,12 dias, com uma fase folicular de 8,8 ± 1,4 dias e 15,62 ± 2,1 dias de fase lútea. Na fase folicular foram observadas secreção sanguinolenta vaginal, edema vulvar, presença de muco e facilidade de introdução do swab, com pico de estrógeno de 240,66 ± 12,69 pg/ml e predomínio de células superficiais na citologia vaginal, com visualização de folículos ovarianos por ultrassonografia. Na fase lútea foi observada ausência de secreção sanguinolenta e dificuldade de introdução do swab, além do aumento nos níveis de progesterona (10,83 ± 1,86 ng/ml) e visualização de corpo lúteo por ultrassom. No segundo experimento, dois protocolos para indução e sincronização de estro foram testados em cutias (Dasyprocta leporina). O primeiro, consistia na administração de duas doses de cloprostenol (5µg) com intervalo de nove dias; e o segundo, na administração de 30 µg de análogo de GnRH, seguida da administração de cloprostenol (5µg) após sete dias e uma nova dose de análogo de GnRH (30µg) após dois dias. Não houve diferença estatística entre os protocolos testados (P=0.1009). Entretanto, a prostaglandina isolada promoveu a indução de estro em 40% das fêmeas, em 3 e 6 dias, e as associações de cloprostenol e GnRH, em 60% das fêmeas, em 4 e 10 dias, após última dose administrada, com pico de estrógeno de 19,95 ± 2,41pg/ml e 9,67 ± 1,8pg/ml, respectivamente. Em ambos os tratamentos, essas fêmeas mostraram características externas de estro como abertura e hiperemia vulvar, muco vaginal e fácil penetração de swab. O terceiro experimento teve por objetivo comparar a eficiência de dois protocolos para sincronização de estro e adaptar a técnica de inseminação artificial (IA) em fêmeas de catetos (Pecari tajacu), sendo este realizado após identificação e aplicação do método de sincronização mais eficaz. O primeiro protocolo baseava-se na administração dupla de cloprostenol (120µg) com intervalo de nove dias, e o segundo, na associação de 400 UI de gonadotrofina coriônica equina e 200 UI de gonadotrofina coriônica humana, em dose única. Para a IA, as fêmeas foram sincronizadas com a associação eCG e hCG. Todas as fêmeas apresentaram estro. Com o uso do cloprostenol, as fêmeas mostraram estro nove dias após a última aplicação da droga, e na associação, após seis dias. Posteriormente à realização da IA, não foram visualizados embriões por ultrassonografia. No entanto, duas fêmeas apresentaram aumento uterino, com presença de líquido anecogênico no lúmen, e níveis altos de progesterona com 30 e 60 dias após a última IA (67.08 ± 16.85 ng/ml e 73.42 ± 22.59 ng/ml, respectivamente). Assim, como conclusão geral, o monitoramento da atividade ovariana em tatus-peba, bem como a indução e sincronização em cutias e catetos, permitem o uso dessas biotécnicas em larga escala, facilitando a gestão dessas espécies em cativeiro.

2
  • HENRIQUE JORGE MAIA COSTA
  • BIOPRODUCT BASED ON COCONUT WATER AND MALTODEXTRINE FOR SHORT TERM PREOPERATIVE FASTING PROTOCOLS

  • Leader : CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • MEMBRES DE LA BANQUE :
  • CRISTIANE CLEMENTE DE MELLO SALGUEIRO
  • DJALMA NUNES MARQUES
  • JOSÉ EDUARDO AGUILAR SIQUEIRA DO NASCIMENTO
  • JOSÉ FERREIRA NUNES
  • RENATO DE AZEVEDO MOREIRA
  • Data: 29 août 2016


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  • The preoperative fasting was instituted many years ago, at the beginning of the anesthetic practice, to ensure the emptying of the stomach contents, avoid bronchial aspiration, vomiting, regurgitation, administration of various drugs to reduce gastric acidity and volume. Preoperative fasting protocols suggest shorter fasting periods especially for liquids, providing more comfort to the patient, and less risk of hypoglycemia and dehydration, without increasing the incidence of perioperative pulmonary aspiration. The aim of the study was to assess whether the bioproduct based on coconut water and maltodextrin (ACP Surgery) supplied the needs of clinical trials of ERAS project (Enhanced Recovery After Surgery) with good tolerance and acceptance. It was conducted a prospective, randomized, controlled, double-blind study in humans with a group of 49 patients at the Santa Casa de Misericordia de Fortaleza, to prove the efficiency and effectiveness of ACP Surgery to achieve the goals and planned effects on ERAS project. The experimental groups were: G1 [control group; Nutri Dextrin (Nutrimed, Fortaleza, Ceará); n = 20) and G2 [intervention; ACP Surgery (ACP Biotecnologia, Fortaleza, Ceará;. N = 29]. In the interval between 10 and 20 hours prior to the surgery, blood sample was taken from each patient and sent to the Hospital's Laboratory of Clinics Analysis for measurement of serum levels of total leukocytes, glucose, sodium, potassium and chlorine. The data were submitted to t test (α = 95%, p> 0.05 and t rejection area > 1.96) and expressed as mean and standard deviation. The G1 (Nutri Dextrin) and G2 (ACP Surgery) showed the following results regarding blood tests: total leukocyte G1 = 11,322.64 + 5,033.94, and G2 = 11,074.14 + 4,480.35; sodium G1 = 140.8 + 4.14 mEq/L, and G2 = 139.3 + 4.38 mEq/L; potassium G1 = 4.16 + 0.65 mEq/L, and G2 = 4.18 + 0.47 mEq/L; chloro G1 = 107.4 + 4.69 mEq/L, and G2 = 107.1 + 6.88 mEq/L; and glucose G1 = 130.3 + 39.96 mg/dL, and G2 = 123.9 + 36.47 mg/dL. In all variables p> 0.05, so it can be considered that Nutri Dextrin and ACP Surgery products are equivalent. The acceptance of the ACP Surgery was 97%, with no reports of cough or nausea (100% tolerance), without loss of palatability. It follows that the produced bioproduct met the needs and specifications of preoperative fasting protocols of short duration, showing acceptance and tolerance compatible with the commercial product at the same time reducing costs and facilitating protocol implement.

3
  • PALOMMA RUSSELLY SALDANHA DE ARAÚJO OLIVEIRA
  • Technological development of respiratory cycle training (RCT) device and evidence on inspiratory muscle training in Chronic Obstructive Pulmonary Disease (COPD)

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • SELMA SOUSA BRUNO
  • VANESSA REGIANE RESQUETI FREGONEZI
  • MARCELO VELLOSO
  • VANESSA SUZIANE PROBST
  • Data: 12 déc. 2016


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  • The respiratory muscles are the cornerstones of ventilation process. Respiratory muscle dysfunctions may be due to respiratory muscle weakness and could contribute to symptoms’ and signals of dyspnea, coughing weakness and exercise intolerance and respiratory failure. Respiratory Muscle Training (RMT) with pressure-loading threshold device has been widely used as adjuvant therapy in healthy individuals to improve respiratory muscles or in patients to restore ventilation. A wide variety of RMT devices are available commercially, but their characteristics are intended for the individual training of muscle groups or have non-specific load adjustments, unviable for athletes and inadequately adjustable for patients with respiratory muscles impairment, such as Chronic Obstructive Pulmonary Disease COPD. Although there is a previous report about evidence of Inspiratory Muscular Training (IMT) in COPD, improperly methodology in the search strategy and low methodological quality of the studies included on previous systematic reviews may contribute to the variety of results about the benefits of this therapy. The objective of this study was to develop a device named Respiratory Cycle Training (RCT), integrated for inspiratory and expiratory muscles, as well as to verify evidence of IMT on dyspnea and exercise capacity in COPD. The methodology involved three research modalities: i) the technological development of the RCT device, ii) the transversal study, and iii) the systematic review associated with the meta-analysis developed together with Cochrane Collaboration. The results corresponded to innovation product applied to health, with invention patent deposit in the INPI (BR 1020160253047); Checking the acute effects of different combinations of respiratory loads on respiratory mechanics in healthy subjects and a scientific paper, accepted on Cochrane Collaboration Library about the effects of IMT in individuals with COPD. The present research will provide support for the development of future studies to investigate a new respiratory muscle training modality, called TCR, with knowledge production in the biotechnology area, as well as contribute to decision making regarding the physiotherapeutic intervention in COPD.

2015
Thèses
1
  • ILLIA NADINNE DANTAS FLORENTINO LIMA
  • Desenvolvimento, avaliação e inovação tecnológica em contração muscular respiratória tixotrópica.

  • Leader : GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • MEMBRES DE LA BANQUE :
  • ARMELE DE FATIMA DORNELAS DE ANDRADE
  • GUILHERME AUGUSTO DE FREITAS FREGONEZI
  • RAQUEL RODRIGUES BRITTO
  • VANESSA REGIANE RESQUETI FREGONEZI
  • WALTER ARAUJO ZIN
  • Data: 10 févr. 2015


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  • O controle da respiração é um processo complexo e multifatorial, responsável pela manutenção da adequada homeostase e suprimento de oxigênio nos tecidos corporais. Os músculos respiratórios são parte primordial deste processo e podem ser treinados para que haja melhora na eficiência na ventilação. São descritas na literatura três modalidades de treinamento, a hiperpneia voluntária isocápnica, o treino resistido a fluxo com válvula e o treino resistido à carga pressórica. Atualmente, merece destaque uma nova modalidade definida como Tixotropia, que envolve contrações inspiratória e expiratória contrauma via aérea ocluída em diferentes volumes pulmonares, como Capacidade Pulmonar Total (CPT) e Volume Residual (VR). O presente estudo teve como objetivo desenvolver e avaliar inovações tecnológicas no âmbito de músculos respiratórios e mecânica respiratória. A pesquisa foi desenvolvida envolvendo o Laboratório de Desempenho Pneumocardiovascular e Músculos Respiratórios da Universidade Federal do Rio Grande do Norte (UFRN) e o Laboratorio Tecnologie Biomedichale - TBM do Politécnico de Milão, Itália. A metodologia adotada foi composta por três itens integrados: (i) depósito de patente de um controlador automático para válvula pneumática para treinamento muscular respiratório na modalidade tixotropia; (ii) revisão sistemática; (iii) artigos científicos relacionados à utilização do instrumento desenvolvido. Os resultados compreenderam depósito de patente no Núcleo de Inovação Tecnológica (NIT) vinculado à UFRN, sob número 23077.077363/2014-59, revisão sistemática sobre testes de endurance com carga constante ou incremental usados em pacientes com Doença Pulmonar Obstrutiva Crônica (DPOC), avaliados pela Pletismografia Optoeletrônica e artigos científicos apresentando resultados dos efeitos agudos da modalidade de treinamento tixotropia e suas repercussões sobre os volumes pulmonares em indivíduos saudáveis utilizando o instrumento desenvolvido.  Portanto, o presente estudo propiciou a criação e validação de instrumento inovador e suas evidências científicas, segundo aspectos da Avaliação de Tecnologias em Saúde (ATS), contribuindo para desenvolvimento de uma nova modalidade e instrumento de treinamento muscular respiratório a ser utilizado em pacientes com doenças que afetam a mecânica respiratória.

     

     

     

     

2
  • THIBÉRIO DE SOUZA CASTELO
  • Obtenção e conservação de espermatozóides de cutia (Dasyprocta leporina Linneaus, 1758) do semiárido brasileiro.

  • Leader : ALEXANDRE RODRIGUES SILVA
  • MEMBRES DE LA BANQUE :
  • ALEXANDRE RODRIGUES SILVA
  • ALEXSANDRA FERNANDES PEREIRA
  • LUCIA DANIEL MACHADO DA SILVA
  • MARIA DENISE LOPES
  • MOACIR FRANCO DE OLIVEIRA
  • Data: 26 févr. 2015


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  • A criopreservação de sêmen é uma importante ferramenta para a conservação e difusão de germoplasma. Para tanto, um protocolo de eletroejaculação adequado é fundamental, para obtenção de espermatozoides viáveis tendo em vista vários fatores interferirem em seu sucesso. Álem disso, apesar dos resultados promissores da criopreservação de espermatozoides epididimários, desconhece-se se seria possível adaptar protocolos de criopreservação de tais espermatozoides para os obtidos por eltroejaculação, principalmente pela inexistência de um protocolo de criopreservação ideal, denotando seu constante aperfeiçoamento. Dessa forma, o objetivo geral da tese é estabelecer protocolos para obtenção e conservação de espermatozoides de cutias (Dasyprocta leporina) criadas em cativeiros no semi-árido brasileiro, visando sua exploração sustentável. A tese foi dividida em três experimentos. No primeiro, foi estudada a influência da interação entre dois tipos de sonda (ondas senoidais e quadráticas) e dois protocolos de estimulação (contínuo e seriado) sobre a eficiência de colheita de sêmen de cutias por eletroejaculação. Assim, a interação mais eficiente na obtenção de ejaculados com espermatozóides foi a que utilizou eletrodos em aneis associado com estímulos em série (4/7; 57%). No segundo, foi realizada a comparação da ação crioprotetora de diferentes substâncias (glicerol, etilenoglicol, dimetilsulfóxido, dimetilformamida) na criopreservação de espermatozóides epididimários. Relativo a isso, os maiores valores de motilidade, vigor e integridade de membrana, foram obtidos nas amostras criopreservadas com glicerol quando comparada as amostras contendo etilenoglicol e dimetilformamida, entretanto, sem diferença (P > 0,05) quando comparada as criopreservadas com dimetilsulfoxido. Por ultimo, foi estudado o efeito dos métodos de obtenção de espermatozoides (eletroejaculação x colheita epididimária) na criopreservação. Assim, não foram observadas diferenças após criopreservação entre os métodos de obtenção de espermatozóides (P≥0,05), denotando a possibilidade de adaptar com sucesso o protocolo de criopreservação de espermatozoides epididimarios para ejaculados de cutias.

3
  • GABRIELA LIBERALINO LIMA
  • CONSERVAÇÃO DE MATERIAL GENÉTICO DE ESPÉCIES SILVESTRES DO BIOMA CAATINGA UTILIZANDO A MANIPULAÇÃO DE OÓCITOS INCLUSOS EM FOLÍCULOS OVARIANOS PRÉ ANTRAIS (MOIFOPA)

  • Leader : ALEXANDRE RODRIGUES SILVA
  • MEMBRES DE LA BANQUE :
  • ALEXANDRE RODRIGUES SILVA
  • ALEXSANDRA FERNANDES PEREIRA
  • LUCIA DANIEL MACHADO DA SILVA
  • MARIA DENISE LOPES
  • MOACIR FRANCO DE OLIVEIRA
  • Data: 27 févr. 2015


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  • O objetivo da presente tese foi utilizar a manipulação de oócitos inclusos em folículos ovarianos pré antrais (MOIFOPA) como ferramenta para o resgate e otimização do uso de gametas femininos oriundos de espécies silvestres do bioma Caatinga. A tese foi dividida em quatro experimentos. No primeiro, foi realizada a estimativa e descrição das características histológicas e ultraestruturas dos folículos pré antrais (FOPA) de cutias (Dasyprocta leporina), nos quais foram estimados 4419.8 ± 532.26 e 5397.52 ± 574.91 folículos para os ovários direito e esquerdo, respectivamente, sendo a maioria (86,63%) pertencente a categoria de folículos primordiais (P<0,05). A maior parte da população consiste de folículos morfologicamente normais (70,78%), apresentando núcleo oocitário grande e central, com citoplasma uniforme. Na avaliação ultraestrutural verificou-se a presença de um grande número de mitocôndrias arredondadas e gotas de lipídios. No segundo experimento, foi realizada a estimativa e a descrição das características dos FOPA de preás (Galea spixii), bem como avaliação do efeito da vitrificação em superfície sólida (VSS) sobre a morfologia de FOPA in situ. O total de 416,0 ± 342,8 FOPA foi estimado por par de ovários e a presença de uma grande quantidade de folículos primários foi evidenciada (P<0,05). A maior parte dos FOPA apresentou-se morfologicamente normal (94,6%), possuindo núcleo oocitário contendo grânulos condensados de heterocromatina. Mitocôndrias com formato arredondado ou alongado representaram as organelas mais abundantes. Quanto a VSS, o protocolo utilizando o dimetilsulfóxido (DMSO) 3M possibilitou a preservação de 69,5% de FOPA morfologicamente normais, sendo evidenciado através da análise por microscopia de luz e eletrônica de transmissão. No terceiro experimento, foi realizada a avaliação do efeito da VSS sobre a morfologia e viabilidade de FOPA in situ de catetos (Pecari tajacu). Não foram observadas diferenças entre os tratamentos, onde o uso dos crioprotetores DMSO, etilenoglicol (EG) e dimetilformamida (DMF), independente da concentração utilizada (3 ou 6 M) promoveu a preservação da morfologia de mais de 70% dos FOPA. Quanto a viabilidade, os crioprotetores DMSO e EG, demonstraram melhor manutenção da mesma. O quarto experimento teve por objetivo avaliar o efeito do α MEM+ ou TCM199 associados ou não a 50 ng de FSHr sobre a morfologia, ativação e crescimento de FOPA de catetos, cultivados in vitro (CIV) durante 1 ou 7 dias e o efeito sobre a matriz extracelular (MEC). Após 7 dias de CIV apenas o TCM199/FSH manteve a proporção de FOPA intactos similar ao dia 1 (63,2%), contudo nenhuma diferença foi observada entre os tratamentos (P>0,05). Adicionalmente, um aumento na proporção de FOPA em desenvolvimento foi verificada (P>0.05). Através da análise com Ag-NOR, observou-se que apenas o tratamento com TCM199/FSH manteve a proporção de proliferação celular similar ao dia 1 (P>0.05). A coloração com picrosirius red revelou que a MEC permaneceu intacta em todos os tratamentos (P>0.05). Assim, como conclusão geral, o uso da MOIFOPA nas referidas espécies permitiu o conhecimento de aspectos relacionados a sua morfofisiologia reprodutiva, possibilitando tanto a conservação do material genético destas espécies, com a possibilidade de formação de bancos de germoplasma, como a elucidação de mecanismos relacionados a sobrevivência e desenvolvimento dos FOPA in vitro.

4
  • GIDEÃO WAGNER WERNECK FELIX DA COSTA
  • MAPEAMENTO FÍSICO DE GENES COM FINS BIOTECNOLÓGICOS EM PEIXES MARINHOS DE INTERESSE COMERCIAL – RACHYCENTRIDAE E LUTJANIDAE

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • WAGNER FRANCO MOLINA
  • KATIA CASTANHO SCORTECCI
  • LUIZ ANTONIO CARLOS BERTOLLO
  • MARCELO DE BELLO CIOFFI
  • MARCELO FRANCISCO DE NÓBREGA
  • Data: 27 févr. 2015


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  • A espécie Rachycentron canadum, conhecida vulgarmente como beijupirá ou cobia, é o único representante da família Rachycentridae, vem sendo crescentemente utilizado na piscicultura marinha, em cultivos intensivos. Como características vantajosas, é de fácil adaptação, prolífica, possui crescimento precoce em cativeiro e elevado valor comercial. Já as espécies da família Lutjanidae (Lutjanus synagris, Lutjanus jocu, Lutjanus analis, Lutjanus alexandrei e Ocyurus chrysurus) representam um importante recurso pesqueiro em todas as áreas de sua ocorrência. No Brasil a exploração comercial dos Lutjanidae se iniciou na década de 60 e nos anos 80, já demonstrava um declínio nos volumes de captura. A diminuição da capturas aponta que os lutjanídeos devem possuir um manejo conservativo. Apesar dos potencias econômicos, pouco se conhece sobre as características genéticas e citogenéticas destas espécies. Principalmente no que diz respeito a análise de DNA repetitivos, representando a maior parte do genoma dos eucariotos, são capazes de desempenharem papeis evolutivos importantes no genoma dos peixes. Dados citogenéticos vem crescentemente sendo empregados em estudos populacionais e com fins biotecnológicos em peixes. As analises citogenéticas foram realizadas utilizando métodos clássicos como coloração com Giemsa, bandamento C e Ag-RONs, coloração com os fluorocromos base-específicos (DAPI e MM) e mapeamento físico de sequências repetitivas dentre as quais, sequências teloméricas, transposons (Tol2), retrotransposons (Rex1 e Rex3), DNA repetitivos (Cot-1 e microssatélites) e das regiões transcricionalmente ativas dos genes ribossomais 18S e 5S e histonas (H2BA e H3), através da hibridação in situ com sondas fluorescentes (FISH). Os padrões cromossômicos obtidos contribuíram para o conhecimento da organização das sequências repetitivas no genoma das espécies, bem como o destacadamente do cariótipo. Padrões incomuns de expansão de sequências histonas retratam a primeira ocorrência em peixes marinhos. Por outro lado, padrões difusos de distribuição de DNA repetitivos propiciarão informações uteis em futuros estudos visando o aumento da produtividade, manejo e incremento do conhecimento genético deste recurso pesqueiro com vistas a subsidiar técnicas de melhoramento genético para a espécie, visando auxiliar o desenvolvimento da piscicultura marinha.
5
  • FRANCISCO CANINDE DE SOUSA JUNIOR
  • Recuperação e purificação do antígeno 503 de Leishmania i. chagasi expresso em E. coli e remoção de endotoxina utilizando adsorção em leito expandido

  • Leader : EVERALDO SILVINO DOS SANTOS
  • MEMBRES DE LA BANQUE :
  • EVERALDO SILVINO DOS SANTOS
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • JORGE DOS SANTOS CAVALCANTI
  • ANA LÚCIA FIGUEIREDO PORTO
  • IVANILDO JOSÉ DA SILVA JUNIOR
  • Data: 27 févr. 2015


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  • O crescente interesse e aplicações dos produtos biotecnológicos vêm aumentando o desenvolvimento de novos processos de recuperação e purificação de proteínas. A adsorção em leito expandido (ALE) tem se destacado como uma técnica promissora para essa finalidade, pois combina em uma única etapa os passos de clarificação, concentração e purificação da proteína alvo, reduzindo assim o tempo e os custos de operação. Neste contexto, o objetivo desta tese foi avaliar a recuperação e purificação do antígeno 503 de Leishmania i. chagasi expresso em E. coli M15 e a remoção de endotoxina por ALE. Para definir as condições ótimas de adsorção e eluição na resina Streamline chelating, foram inicialmente realizados ensaios em taques agitados sob a forma de dois planejamentos experimentais. Nos ensaios de adsorção usando o leito na forma expandida empregou-se uma coluna de 2,6 cm de diâmetro por 30,0 cm de altura, acoplada a uma bomba peristáltica. Na segunda etapa do trabalho, o tensoativo não-iônico Triton X-114 foi adicionado à etapa de lavagem da ALE visando a remoção de endotoxina durante o processo de purificação. Buscou-se também a elaborar um modelo matemático capaz de prever as curvas de ruptura do antígeno 503 em coluna na forma expandida. Os resultados do planejamento experimental para adsorção do antígeno 503 mostraram o pH 8,0 e a concentração de NaCl 2,4 M como melhores condições adsorção. No segundo planejamento, o único fator significativo para eluição foi a concentração de imidazol, definida em 600 mM. A isoterma de adsorção do antígeno 503 mostrou bom ajuste ao modelo de Langmuir (R=0,98) e os valores de qmax (capacidade máxima de adsorção) e Kd (constante de equilíbrio) estimados foram de 1,95 mg/g de adsorvente e 0,34 mg/mL, respectivamente. Através dos testes de purificação diretamente do homogeneizado não clarificado obteve-se uma recuperação de 59,2% da proteína de interesse e um fator de purificação de 6,0.  A adição do tensoativo não-iônico Triton X-114 à etapa de lavagem da ALE proporcionou altos valores (>99%) de remoção do LPS inicialmente presente nas amostras para todas as condições estudadas. Na etapa de modelagem matemática da adsorção do antígeno 503 em coluna na forma expandida, as curvas de ruptura experimentais permitiram determinar-se as eficiências do processo (15% de saturação) e da coluna (85% de saturação), no entanto o modelo apresentou bom ajuste apenas para eficiência do processo. O modelo validado foi utilizado com uma estratégia híbrida na otimização da eficiência do processo, indicando que os valores de 7,8 cm de altura inicial do leito, 248,8 cm/h de velocidade superficial e 0,338 g/L de concentração inicial do antígeno resultam no valor de 89,19% para eficiência do processo.

6
  • FRANCISCO HUMBERTO XAVIER JUNIOR
  • Sistemas dispersos para administração oral de paclitaxel baseados em microemulsão e nanocápsulas mucoadesivas contendo o óleo de copaíba

  • Leader : ERYVALDO SOCRATES TABOSA DO EGITO
  • MEMBRES DE LA BANQUE :
  • CHRISTINE VAUTHIER
  • ERYVALDO SOCRATES TABOSA DO EGITO
  • Gilles Ponchel
  • PATRICK SAULNIER
  • SYLVIE CRAUSTE-MANCIET
  • Data: 5 mars 2015


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  • O câncer é uma das doenças mais importantes do mundo, tanto pelo aumento da sua incidência, como da sua prevalência e mortalidade. O paclitaxel é um dos medicamentos anticancerígenos mais potentes atualmente utilizados para o tratamento do câncer de pulmão, mama, ovário, carcinomas cervicais de útero, cérebro, leucemia aguda, entre outros. Este fármaco apresenta uma estrutura de pseudoalcaloide com uma estrutura diterpenoide. Esta molécula extraída de Taxus brevifolia possui mecanismo de ação pela promoção da estabilização de microtúbulos, inibindo a proliferação célular e como consequência a indução da apoptose. Como inúmeros agentes anticancerígenos, o paclitaxel é administrado por via intravenosa devido a sua baixa biodisponibilidade oral. No entanto, a via oral suscita um interesse crescente para a administração de medicamentos anticancerígenos. Mesmo diante da potencialidade desta via, a mesma é limitada por problemas relacionados as propriedades físico-químicas do fármaco, fatores fisiológicos e as formas farmacêuticas que reduzem a biodisponibilidade oral do medicamento. Para superar essas limitações, nos últimos anos, sistemas lipídicos e poliméricos nanoparticulados para administração de fármacos têm sido utilizados. Entre estes sistemas, as microemulsões tem demonstrado um melhor potencial de solubilização de moléculas fracamente solúveis e lipofílicas, além de melhorar a estabilidade e proteção destes fármacos contra a degradação. Já entre os sistemas poliméricos, as nanocápsulas biodegradáveis são sistemas vesiculares, em que o principio ativo pode ser confinado em uma cavidade formada por um núcleo líquido, indicado para controlar a libertação do fármaco em sítios alvos, além de melhorar a absorção dos mesmos. O objetivo deste trabalho foi desenvolver sistemas dispersos para via oral, contendo na fase interna óleo de copaíba servindo de veículos para fármacos anticancerígenos, como paclitaxel. Os estudos foram desenvolvidos em dois sentidos com o objetivo de formular adequadas microemulsões e nanocápsulas mucoadesivas. Na primeira parte desta tese foi desenvolvido um método de análise e doseamento de óleo de copaíba por cromatografia gasosa com detector de massas. Posteriormente, um método de dosagem de paclitaxel no óleo de copaíba foi desenvolvido e validado a fim de determinar sua solubilidade e coeficiente de partição utilizando cromatografia líquida de alta eficiência. Os principais compostos identificados no óleo essencial de copaíba foram β-bisaboleno (23,6%), β-cariofileno (21,7%) e α-bergamoteno (20,5%). Para o óleo resina de copaíba foram o ácido copálico (15,6%), β-bisaboleno (12,3%), β-cariofileno (7,9%), α-bergamoteno (7,1%) e ácido Labd- 8 (20) -eno-ácido 15,18-dióico (6,7%). O método desenvolvido para a dosagem do óleo de copaíba foi linear (superior a r²=0,99) para intervalo de concentração entre 40 a 160 µg/mL. O limite de detecção foi determinado como sendo 6,38, 4,16 e 0,55 µg/mL e o limite de quantificação foi de 19,36, 12,62 e 1,67 µg/mL para o β-cariofileno, α-humuleno e óxido de cariofileno, respectivamente. A precisão e exatidão foram inferiores a 4,4 e 3,5%, respectivamente. Para a quantificação de paclitaxel, o método foi considerado linear (r² = 0,9998) para um intervalo de concentração de 50-2000 ng/mL e apresentou resíduos da dispersão homocedástico. Os limites de detecção e de quantificação para o paclitaxel foram de 5,54 e 1,85 ng/mL, respectivamente. As determinações de exatidão e precisão foram inferiores ou iguais a 0,82 e 0,65%, respectivamente. A solubilidade de paclitaxel em óleo resina de copaíba foi de 0,8 mg.mL-1 e o valor de coeficiente de partição foi 3. Posteriormente, microemulsão de óleo de copaíba em água foi desenvolvida em função dos cálculos dos parâmetros de solubilidade entre os componentes do óleo de copaíba e os surfactantes. Tal processo levou à obtenção de microemulsão estável com o óleo essencial de copaíba cotendo elevada fração volumétrica da fase dispersa (19,6%) e uma baixa concentração de surfactante (13,7%). Esta formulação permitiu a incorporação de paclitaxel a uma concentração de 0,37 mg/mL, sem provocar perturbação notáveis no tamanho e potencial zeta das gotículas das microemulsões. Em seguida, o estudo de mucoadesão das microemulsões contendo paclitaxel radiomarcado permitiu verificar que cerca de 4% do fármaco encontrou-se ligado a mucosa intestinal de ratos. O segundo sistema desenvolvido neste trabalho consistiu de nanocápsulas mucoadesivas para encapsulação do óleo de copaíba. A formulação foi baseada na abordagem de planejamento experimental, utilizando três variáveis independentes (pH, temperatura e concentração de quitosana no meio) e parâmetros de resposta de tamanho e o potencial zeta das nanocápsulas. Os sistemas otimizados desenvolvidos apresentaram um diâmetro de 473 nm, o potencial zeta de +34 mV e a eficiência de encapsulação do óleo de copaíba foi determinada em 75,8%, correspondendo a 55,5 mg de β- cariofileno / mg de nanocápsulas. Durante o processo de encapsulação não houve alteração físico-química dos compostos presentes no óleo de copaíba. O paclitaxel foi incorporado sem alteração do tamanho, morfologia e o potencial zeta das nanocápsulas. A encapsulação de paclitaxel foi de 75%, o que corresponde a 23 µg/mg de polímero. O desenvolvimento deste sistema ainda incluiu a marcação com uma sonda fluorescente e incorporação de paclitaxel radioativo. Esta formulação foi estável em meio gástrico durante 120 minutos e depois de seis meses a 4 °C. Quanto ao estudo mucoadesão, 9% de nanocápsulas desenvolvidos foram aderidas a mucosa intestinal, o que corresponde a uma razão de 3,35 g de nanocápsulas/m² da mucosa intestinal. Os resultados deste trabalho conduziram ao desenvolvimento de duas formulações de paclitaxel em nanosistemas originais contendo óleo natural de copaíba que estão prontos para avaliação da sua capacidade de entregar de fármaco anticancerígeno pela via oral.


7
  • NATHALIA KELLY DE ARAUJO
  • Produção e purificação de enzimas quitosanolíticas produzidas por microrganismo isolado no Nordeste Brasileiro.

  • Leader : EVERALDO SILVINO DOS SANTOS
  • MEMBRES DE LA BANQUE :
  • EVERALDO SILVINO DOS SANTOS
  • CRISTIANE FERNANDES DE ASSIS
  • MATHEUS DE FREITAS FERNANDES PEDROSA
  • ELIANA SETSUKO KAMIMURA
  • MICHELLE ROSSANA FERREIRA VAZ
  • Data: 23 mars 2015


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  • Uma cepa produtora de quitosanase foi isolada e identificada como Bacillus cereus C-01. A partir disso a purificação da enzima e sua caracterização bioquímica foram estudadas. Para purificação foi utilizado a resina da linha streamline Dietilaminoetil (DEAE) e coluna de vidro (2,6 cm x 30,0 cm) em sistema de Adsorção em Leito Expandido (ALE).  As condições para a purificação da quitosanase foram otimizadas estatisticamente através de um Planejamento Composto Central. As variáveis que influenciaram significativamente os parâmetros fator de purificação (P) e rendimento da enzima (Y) foram determinadas. As análises estatísticas mostraram que as melhores condições para o máximo P foram 150 cm.h-1 (velocidade de aplicação da amostra/carga), 6,0 cm de altura do leito fixo e 7,36 cm de altura do distribuidor. Em condições otimizadas, a enzima mostrou ligar-se a resina na mesma medida usando caldo clarificado e não clarificado (0,32 e 0,3 U/g adsorvente, respectivamente). A fração recuperada após a eluição exibiu 31% de rendimento com um aumento de 1,35 vezes na atividade específica em relação ao inicial. Se analisada apenas fração eluída com NaCl 0,3 M é possível encontrar um fator de purificação de 1,75. A enzima purificada apresentou estabilidade entre as temperaturas de 30-55ºC durante 60 minutos, pH de 5-8 por 24 horas, e apresentou máxima atividade em pH 5,5 e temperatura de 55ºC. Os íons de Cu2+, Fe2+ e Zn2+ foram inibitórios para a quitosanase. Em contraste, a enzima foi significativamente ativada por Mn2+. Os resultados demonstraram que é possível purificar múltiplas proteínas, a partir de um extrato bruto sem qualquer pré-tratamento, com um processo de purificação econômico e de um único passo.

     

2014
Thèses
1
  • FABIANO AVELINO GONÇALVES
  • Avaliação do potencial da fibra e casca de coco maduro, casca de coco verde e cacto pré-tratados visando à produção de etanol.

  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • GORETE RIBEIRO DE MACEDO
  • CRISTIANE FERNANDES DE ASSIS
  • JACKSON ARAUJO DE OLIVEIRA
  • ANA LÚCIA FIGUEIREDO PORTO
  • JOSÉ ANTÔNIO COUTO TEIXEIRA
  • MARIA VALDEREZ PONTE ROCHA
  • Data: 23 avr. 2014


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  • As condições ambientais influenciam diretamente a qualidade de vida dos seres vivos e as ações antrópicas geralmente modificam as condições ambientais de forma negativa, como exemplo, o aquecimento global, promovido pelas emissões de gases do efeito estufa, especialmente por dióxido de carbono gerado principalmente pelos produtos à base de petróleo e derivados. Uma opção para mitigar essas gerações de gases consiste no uso de biocombustíveis, pois além de serem menos poluentes, são renováveis. Nesse contexto, o presente trabalho investigou o potencial de diferentes materiais lignocelulósicos (fibra de coco maduro, casca de coco verde e casca de coco maduro), resíduos gerados em regiões rurais e urbanas, além de vegetal cultivado em ambientes inóspitos (cacto), sendo todos, materiais abundantes na região Nordeste do Brasil. Esses materiais foram submetidos aos pré-tratamentos com peróxido hidrogênio alcalino seguido por hidróxido de sódio (PHA-PHS), autohidrólise (PA), hidrotérmico catalisado com hidróxido de sódio (PHCHS) e organosolv usando etanol alcalino (POEA). Os materiais pré-tratados foram submetidos à hidrólise enzimática e nas estratégias de fermentação e sacarificação simultânea (SSF) e fermentação e sacarificação semi-simultânea (SSSF), utilizando-se Saccharomyces cerevisiae, Pichia stipitis e Zymomonas mobilis. Avaliou-se também, a presença de compostos inibitórios (hidroximetilfurfural, furfural, ácido acético, ácido fórmico e ácido levulínico) e a água marinha presente no meio de cultivo. Os materiais pré-tratados por PHA-PHS resultaram na deslignificação dos materiais em uma faixa entre 54,0 e 71,0%, contendo entre 51,80 e 54,91% de celulose, entre 17,65 e 28,36% de hemicelulose, entre 7,99 e 10,12% de lignina. As hidrólises enzimáticas proporcionaram conversões dos materiais em glicose entre 68,0 e 76,0%. Os rendimentos em etanol utilizando a SSF e SSSF para a fibra de coco maduro pré-tratada variaram entre 0,40 e 0,43 g/g, 0,43 e 0,45 g/g, respectivamente. Os materiais pré-tratados por PA apresentaram rendimentos de sólidos entre 42,92 e 92,74%, contendo entre 30,65 e 51,61% de celulose e entre 21,34 a 41,28% de lignina. As hidrólises enzimáticas resultaram nas conversões dos materiais em glicose entre 84,10 e 92,52%. Os rendimentos em etanol utilizando casca de coco verde pré-tratada e as estratégias SSF e SSSF foram entre 0,43 e 0,45 g/g. A fibra de coco maduro pré-tratada por PHCHS apresentou rendimentos de sólidos entre 21,64 e 60,52%, com aumento de celulose entre 28,40 e 131,20%, redução de hemicelulose entre 43,22 e 69,04%, redução de lignina entre 8,27 e 89,13%. A hidrólise enzimática resultou em conversão do material em glicose de 90,72%. Os rendimentos em etanol utilizando a SSF e SSSF foram 0,43 e 0,46 g/g, respectivamente. Os materiais pré-tratados por POEA apresentaram reduções de sólido entre 10,75 e 43,18%, aumento de celulose em até 121,67%, redução de hemicelulose em até 77,09% e redução de lignina em até 78,22%. As hidrólises enzimáticas resultaram nas conversões dos materiais em glicose entre 77,54 e 84,27%. Os rendimentos em etanol utilizando cacto pré-tratado e as estratégias SSF e SSSF foram entre 0,41 e 0,44 g/g, 0,43 e 0,46 g/g, respectivamente. As fermentações alcoólicas realizadas em biorreator resultaram em rendimentos e produção de etanol entre 0,42 e 0,46 g/g, 7,62 e 12,42 g/L, respectivamente. Os compostos inibitórios apresentaram maiores efeitos sinérgicos negativos nas fermentações realizadas por P. stipitis, Z. mobilis e S. cerevisiae, respectivamente. O ácido fórmico e ácido acético apresentaram efeitos mais significativos entre os compostos inibitórios, seguido pelo hidroximetilfurfural, furfural e ácido levulínico. As fermentações realizadas em meio de cultivo diluído com água marinha apresentaram resultados promissores, especialmente quando se utilizou S. cerevisiae (0,50 g/g) e Z. mobilis (0,49 g/g). Os diversos resultados obtidos no presente trabalho indicam que, os materiais lignocelulósicos, pré-tratamentos, estratégias fermentativas e micro-organismos estudados merecem atenção por parecerem promissores e passíveis de serem utilizados no contexto de biorrefinaria, visando à produção de etanol.  

2013
Thèses
1
  • LEONAM GOMES COUTINHO
  • Caracterização da proteína APE1/ref-1 como alvo terapêutico na redução do estresse oxidativo gerado pela resposta inflamatória.

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • CARLOS RENATO MACHADO
  • EDDA LISBOA LEITE
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • SELMA MARIA BEZERRA JERONIMO
  • TIRZAH BRAZ PETTA
  • Data: 19 mars 2013


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  • A meningite bacteriana (MB) é uma doença infecciosa que permanece com altas taxas de mortalidade e morbidade em todo o mundo, principalmente em países subdesenvolvidos, apesar dos avanços na antibioticoterapia. Um dos principais mecanismos associados às sequelas durante a MB é a elevada resposta inflamatória, que promove uma exacerbada quantidade de espécies reativas de oxigênio (ERO) levando às células a apoptose ou necrose. A ativação de enzimas de reparo, durante o estresse oxidativo tem sido demonstrada nas mais diversas desordens, entre elas a endonuclease apurínica/apirimidínica 1/ efetor redox 1 (APE1/ref-1). APE1/ref-1 é uma enzima multifuncional envolvida no reparo de DNA e na redução de fatores envolvidos com a resposta inflamatória, tais como o fator nuclear kappa B (NFκB) e proteína ativadora 1 (AP-1). Portanto, este trabalho teve como objetivo buscar novos alvos terapêuticos para a redução de sequelas durante MB. Para isto, inicialmente foi realizado uma análise no perfil de expressão de citocinas em líquor de pacientes com meningite causada por Streptococcus pneumoniae e Neisseriae meningitidis. Em seguida, foi avaliado o papel da APE1 como um possível alvo terapêutico na modulação das citocinas expressas em monócitos durante resposta celular a lipopolissacarídeo (LPS). Por fim, foi observado a expressão de APE1 no córtex (CX) e hipocampo (HC) de ratos com MB frente a terapia adjuvante com vitamina B6. Nosso estudo mostrou principalmente que interferon gama (IFNγ) é significativamente mais expressa em pacientes com S. pneumoniae do que N. meningitidis. Por outro lado, o uso de inibidores das funções redox e de reparo de APE1 reduzem a expressão de citocinas, principalmente o fator de necrose tumoral-alfa (TNF-α), bem como, o nível de ERO nas células estimuladas com LPS. Ainda, a expressão protéica de APE1, no CX e HC dos ratos, foi modulada após introdução da vitamina B6. Estes dados fornecem um novo olhar para a fisiopatologia da MB, em que citocinas como IFNy podem ser usadas em um diagnóstico diferencial, enquanto que APE1 pode ser usada como alvo para modular a resposta inflamatória e a vitamina B6 pode ser usada para modular APE1. Consequentemente, o conhecimento obtido neste estudo pode ser importante na melhoria do prognóstico da MB, além de contribuir para entender a associação entre o reparo de DNA e inflamação.

2
  • THAYSE AZEVEDO DA SILVA
  • Análise do efeito de polimorfismos não-sinônimos em genes de reparo de DNA da via BER na resposta inflamatória da Meningite.

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • ANTONIO CARLOS ROSÁRIO VALLINOTO
  • JENIFER SAFFI
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • SANDRO JOSE DE SOUZA
  • TIRZAH BRAZ PETTA
  • Data: 21 mars 2013


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  • Estudos in vitro e em modelos animais sugerem que as proteínas de reparo de DNA da via BER APE1, OGG1 e PARP-1 estão envolvidas também na resposta inflamatória. Neste trabalho foi investigado se os SNPs APE1 Asn148Glu, OGG1 Ser326Cys e PARP-1 Val762Ala associam-se à meningite e desenvolvido um sistema para análise funcional destas variantes polimórficas. Os genótipos de pacientes com meningite bacteriana (MB), meningite asséptica (MA) e não  infectados (controles) foram investigados por PIRA-PCR ou PCR-RFLP. Danos no DNA foram detectados no DNA genômico por meio de tratamento com FPG. IgG e IgA foram titulados no plasma e citocinas e quimiocinas foram mensuradas em  amostras de líquor através de ensaios em Bio-Plex. Os níveis de NF-κB e c-Jun foram dosados no líquor dos pacientes por meio de dot blot. Foi observado um aumento significativo (P<0.05) na frequência do alelo APE1 Glu nos casos de MB e MA. Os genótipos Asn/Asn no grupo controle e Asn/Glu no grupo da MB também apresentaram relevante aumento em suas frequências (P<0.05). Para o SNP OGG1 Ser326Cys, o genótipo Cys/Cys esteve mais frequente (P<0.05) nos casos de MB. A  frequência do genótipo PARP-1 Val/Val foi mais alta no grupo controle (P<0.05). A  ocorrência combinada dos SNPs foi significativamente alta nos pacientes com MB, indicando que estes SNPs podem estar associados à doença. Os portadores do alelo APE1 Glu ou OGG1 Cys apresentaram um número maior de sítios sensíveis à FPG, sugerindo que os SNPs afetam a atividade de rep aro do DNA. Alterações na síntese de IgG foram observadas na presença dos SNPs APE1 Asn148Glu, OGG1 Ser326Cys ou PARP-1 Val762Ala. Reduções nos níveis de IL-6, IL-1Ra, MCP-1/CCL2 e IL-8/CXCL8 foram encontradas na presença do alelo APE1 Glu em  amostras de pacientes com MB, no entanto não foram encontradas diferenças nos níveis de NF-κB e c-Jun considerando os genótipos e os grupos analisados. Utilizando APE1 como modelo, foi desenvolvido um sistema que possibilita a expressão e caracterização funcional das enzimas polimórficas estudadas e seus efeitos na célula, por meio de clonagem, utilizando o vetor pIRES-EGFP e cDNA de APE1, transfecção celular da construção obtida, inibição por siRNA de APE1 endógena e genotipagem de culturas celulares hospedeiras. Em conclusão, foram obtidas evidências de um efeito significativo dos SNPs nos genes de reparo de DNA na regulação da resposta imunológica. Este é um trabalho pioneiro na área, que demonstra a associação de variantes das enzimas da via BER com uma doença infecciosa em humanos, sugerindo que os SNPs estudados podem afetar a resposta imune e o nível de estresse oxidativo durante a infecção cerebral. Desta forma, novos meios de análise funcional devem ser desenvolvidos para estudo de proteínas polimórficas e suas interações neste contexto.

3
  • JOANA CRISTINA MEDEIROS TAVARES MARQUES
  • Transcriptoma diferencial entre células-tronco mesenquimais humanas jovens e senescentes.

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • DEBORAH AFONSO CORNELIO
  • GUIDO LENZ
  • JOSÉ MIGUEL ORTEGA
  • SANDRO JOSE DE SOUZA
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • Data: 25 mars 2013


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  • Células-tronco mesenquimais humanas (CTMH) são muito úteis na terapia celular. O longo período de cultivo pode resultar em senescência replicativa ou estar relacionado com o aparecimento de alterações cromossômicas responsáveis pela aquisição de um caráter tumorigênico in vitro. Neste estudo, foi comparado o transcriptoma de CTMH jovens e senescentes obtidas de diferentes doadores. Além disso, pela primeira vez, o perfil de expressão de CTMH com uma inversão cromossômica paracêntrica (CTMH/inv) foi comparado ao de CTMH que possuem cariótipo normal (CTMH/n) em passagens jovens e senescentes de cultivo in vitro. As CTMH utilizadas neste estudo foram isoladas da veia do cordão umbilical de três dadores, dois CTMH/n e de um CTMH/inv. Após a criopreservação, elas foram expandidas in vitro até alcançarem a senescência. O RNA total foi extraído utilizando o RNeasy mini kit (Qiagen), marcado, purificado e fragmentado com o ® 3 'GeneChip IVT expresso Kit (Affymetrix, Inc.). Subsequentemente, o RNA fragmentado foi hibridado no microarranjo Affymetrix Human Genome U133 Plus 2.0 (Affymetrix, Inc.). A análise estatística da expressão diferencial foi realizada usando o Partek Suite Software Genomic, versão 6.4 (Partek, Inc.). Foram consideradas estatisticamente significativas as diferenças na expressão com valor de P ˂0.01 corrigido com Bonferroni. Apenas os sinais com fold change ˃3.0 foram incluídos na lista de diferencialmente expressos. Diferenças na expressão gênica observadas no estudo dos microarranjos foram confirmadas por resultados de RT-PCR em tempo real. Para a interpretação biológica dos dados foram utilizados: IPA (Ingenuity Systems) para análise de enriquecimento de funções; STRING 9,0 para a construção de redes de interações; Cytoscape 2,8 para a visualização das redes e análises de bottlenecks com o auxílio do software GraphPad Prism 5.0. O pluggin BiNGO do Cytoscape foi utilizado para avaliar a representação de categorias funcionais no Gene Ontology nas redes biológicas. A comparação entre senescentes e jovens em cada grupo de CTMH mostrou que há uma diferença no perfil de expressão, sendo maior nas senescentes do grupo CTMH/inv. Os resultados também mostraram que há diferença nos perfis de expressão entre as CTMH/inv e CTMH/n, sendo maior a diferença quando as células estão senescentes. Novas redes foram identificadas para genes relacionados com a resposta ao longo do tempo de cultivo nos dois grupos de CTMH. Foram identificados genes que podem coordenar funções importantes mais enriquecidas nas redes, como por exemplo, CXCL12, SFRP1,  EGF, SPP1, MMP1 e THBS1. A interpretação biológica destes dados sugere que a população de células CTMH/inv tem diferentes características constitucionais, relacionadas com o seu potencial de proliferação, diferenciação e resposta a estímulos, responsáveis por um processo de senescência replicativa em CTMH/inv  distinto das CTMH/n. Os genes identificados neste estudo são candidatos a marcadores da senescência celular em CTMH, mas a sua relevância funcional neste processo deve ser testada em experiências adicionais in vitro e/ou in vivo

4
  • INGRID VILAR ACCIOLY
  • Processo Biotecnológico voltado a Produção de Estoques Poliplóides do Camarão Litopenaeus vannamei.

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • CIBELE SOARES PONTES
  • JOSE VERISSIMO FERNANDES
  • KARINA RIBEIRO
  • TATJANA KEESEN DE SOUZA LIMA CLEMENTE
  • WAGNER FRANCO MOLINA
  • Data: 25 mars 2013


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  • A melhoria genética de plantéis reprodutores, baseada no melhoramento clássico tem sido tentada, mas sem resultados práticos, até o momento. Protocolos biotecnológicos utilizados em organismos aquáticos cultivados que visam aumentar as taxas de crescimento e resistência a doenças, vêm sendo estudados e aperfeiçoados. Entre as técnicas disponíveis, a aplicação de manipulação cromossômica, embora ainda incipiente, se apresenta como ferramenta voltada a mitigar questões ecológicas e econômicas na carcinicultura. A poliploidização artificial, método já empregado em peixes e moluscos, vem sendo largamente pesquisado para o uso em camarões cultivados. Algumas limitações da expansão deste método em camarões se referem a um melhor conhecimento de aspectos citogenéticos, do nível de dimorfismo sexual e performance em condições de cultivo. Visando contribuir sobre algumas destas questões, a presente pesquisa visou à caracterização citogenética das espécies Litopenaeus vannamei (Decapoda) e Artemia franciscana (Anostraca), analisar a eficácia de métodos de detecção da ploidia, através do uso da citometria de fluxo em processos de indução da poliploidia por choque térmico frio, em diferentes fases do desenvolvimento de ovos recém-fertilizados. Adicionalmente, visou, ainda, a comparação qualitativa e quantitativa do desenvolvimento larval entre organismos diploides e poliplóides, além da identificação do dimorfismo sexual em L. vannamei,por meio da morfometria geométrica. Os resultados obtidos propiciam informações relevantes ao aprimoramento e difusão do uso aplicado de métodos biotecnológicos voltados para a produtividade na carcinicultura nacional.

5
  • UASKA BEZERRA E SILVA
  • Análise metagenômica da microbiota de ambientes aquáticos do estado do Rio Grande do Norte - Brasil.

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • ADRIANA FERREIRA UCHOA
  • RAQUEL CORDEIRO THEODORO
  • BETANIA FERRAZ QUIRINO
  • CLAUDIA ELIZABETH THOMPSON
  • Data: 16 avr. 2013


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  •           A busca por genes baseado na construção e análise de bibliotecas metagenômicas a partir de solo gera oportunidades para explorar uma enorme diversidade genética  e metabólica de microrganismos. Os rios são ecossistemas com alta diversidade  biológica mais pouco explorado por meio de metagenomas. Com esse objetivo, uma biblioteca metagenômica foi construída a partir de DNA extraído de substrato do rio em três pontos ao longo do rio Jundiaí-Rio Grande do Norte. Os pontos de amostragem são derivados de área aberta, terreno acidentado e com a incidência direta da luz solar. O solo é úmido, compacto, de cor escura e coberto com finas camadas de sal. O rio é caracterizado por drenagem intermitente e alta salinidade, que pode ser superior a quatro vezes a concentração do mar. Esta biblioteca foi analisada funcionalmente e também com base em sequências. Para a análise funcional, foi utilizado meio de  cultura sólido de Luria-Bertani (LB) com a concentração de NaCl variando de 0,17M a 0,85M. Foram obtidos 15 clones positivos com características halotolerantes. Os DNAs recombinantes foram extraídos e retransformados em DH10B e curvas de sobrevivência foram obtidas para confirmação e quantificação da resistência ao estresse abiótico. As sequências dos clones foram obtidas e submetidas a ferramenta BLASTX e assim foi comprovado que alguns clones codificavam proteínas hipotéticas. Um dos clones apresentou uma  ORF completa com elevada similaridade de glucose-6-fosfato-isomerase que participa na síntese do percurso de glicerol e serve como um soluto compatível para equilibrar a pressão osmótica no interior e no exterior das células. Posteriormente, com o intuito  de identificar genes que codificam osmolitos relacionados com a halotolerância, amostras de DNA ambiental do substrato do rio e da coluna d´água do estuário e oceano foram piro sequenciadas.sequências de osmolitos de diferentes microrganismos foram obtidas a partir do UniProt e usado como RefSeqs para a identificação por homologia (TBLASTN) nos bancos de dados. As sequências foram submetidas a HMMER para a identificação de domínios funcionais e sequências não redundantes para evitar a quantificação repeti da de um mesmo microrganismo e permitir o agrupamento dos dados por metagenoma. Foram identificados trealose, ectoine, sorbitol, glicerol, inositol-3-fosfato desidrogenase, chaperone, a betaína de glicina e prolina oxiredutase. Estes são osmólitos comumente relacionados a ambientes salinos, no entanto, é aceitável que microrganismos presentes no rio possam produzir esses solutos devido a alta concentração de sal no solo e na água durante a estação de seca neste rio. Isso confirma a existência de uma resposta especializada contra o estresse salino por bactérias no ambiente do rio Jundiaí.

6
  • VALERIA NOGUEIRA DA SILVA
  • Interação de microrganismos na solubilização de fósforo e potássio de rochas para produção de biofertilizantes

  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • GORETE RIBEIRO DE MACEDO
  • EVERALDO SILVINO DOS SANTOS
  • MARIA GIOVANA BINDER PAGNONCELLI
  • MARIA DO CARMO CATANHO PEREIRA DE LYRA
  • MICHELLE ROSSANA FERREIRA VAZ
  • Data: 3 mai 2013


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  • A agricultura atual se baseia no uso intensivo de fertilizantes industrializados, por sua resposta rápida, porém traz consequências danosas ao ambiente, e faz-se necessário o uso de insumos modernos. Uma alternativa é o uso de biofertilizantes de rochas na agricultura, um produto de fácil manuseio, com efeito residual maior e não agride o meio ambiente. O objetivo do estudo foi avaliar a inoculação e co-inoculação de diferentes microrganismos na solubilização de fósforo e potássio de rochas moídas avaliando o melhor desempenho microbiano(s) na produção de biofertilizantes comparando com as rochas puras nas propriedades químicas do solo e, verificar o efeito da inoculação da bactéria Paenibacillus polymyxa na absorção dos minerais solubilizados no desenvolvimento do feijão caupi (Vigna unguiculata [L.] Walp.). O primeiro bioensaio foi conduzido em Laboratório (UFRN) por 72 dias em Placas de Petri, onde o pó de rocha era acrescido de 10% de enxofre e inoculados e co-inoculados com suspensão bacteriana de Paenibacillus polymyxa cultivada em meio caldo triptona de soja, Ralstonia solanacearum em meio Kelman, Cromobacterium violaceum em meio Luria-Bertani e Acidithiobacillus thiooxidans em meio Tuovinen e Kelly, e os fungos Penicillium fellutanum e Tricoderma humatum em meio contendo extrato de malte. A cada 12 dias, amostras eram retiradas a fim de construir uma curva de liberação dos minerais. O segundo bioensaio foi conduzido em casa de vegetação da Empresa de Pesquisa Agropecuária do Rio Grande do Norte, onde utilizou-se 10 kg do Argissolo Amarelo Distrófico, por vaso, com adição dos tratamentos super fosfato simples (SS), cloreto de potássio (KCl), rocha pura, biofertilizantes nas doses 40, 70, 100 e 200% da recomendação para SS e KCl, e uma testemunha, inoculados ou não com a bactéria P. polymyxa. Foram utilizadas sementes da caupi BRS Potiguar e co-inoculadas com suspensão bacteriana de Bradyrhizobium japonicum e P. polymyxa. A primeira colheita foi aos 45 dias de plantio avaliando a matéria seca da parte aérea (MSPA), teores de macronutrientes (N, P, K, Ca, Mg,) e micronutrientes (Zn, Fe, Mn) na MSPA. A segunda colheita aos 75 dias avaliando teores de macro e micronutrientes na planta e no solo, e a capacidade máxima de adsorção de P (CMAP) no solo. Os resultados mostraram que houve sinergismo nas co-inoculações com P. polymyxa+R. solanacearum e, P. polymyxa+C. violaceum com maiores solubilizações de K e P, respectivamente, e melhor tempo de solubilização aos 36 dias. O pH foi mais reduzido nos biofertilizantes de maiores doses, e houve melhoras com sua adição para P na maior dose. Houve redução significativa da CMAP com o aumento da dose do biofertilizante. Houve efeito da fertilização na absorção, com melhoras para P, K e MSPA com SS+KCL, e N, Ca e Mg para biofertilizantes. De modo geral, o P. polymyxa não influenciou na absorção dos elementos na planta. Para Ca e K houve melhoras com SS+KCl, e para Mg com rocha pura. O fertilizante químico foi significativamente superior para peso e número de grãos, sem a presença do P. polymyxa. Na presença da bactéria, biofertilizantes e fertilizante químico apresentaram valores positivos em relação à rocha e testemunha. Os dados evidenciaram que as rochas e os biofertilizantes podem suprir a necessidade de nutrientes às plantas revelando-se como potencial para uma agricultura sustentável.

7
  • LEONARDO LUIZ CALADO
  • Mapeamento cromossômico de genes ribossomais em espécies estuarinas da família Gerreidae – Identificação de uniformidade cariotípica e sua empregabilidade com fins biotecnológicos - REPROGEN.

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • WAGNER FRANCO MOLINA
  • GEORGE SHIGUEKI YASUI
  • JOSE GARCIA JUNIOR
  • LILIANE DE LIMA GURGEL
  • MARCELO FRANCISCO DE NÓBREGA
  • Data: 19 juil. 2013


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  • Os Perciformes são dominantes no ambiente marinho, caracterizados como a maior e mais diversificada Ordem de peixes dentre os teleósteos. Muitas de suas famílias, como os Gerreidae, conhecidos popularmente como carapicus, carapebas, ou mojarras, têm um alto potencial econômico, no que se diz respeito à piscicultura marinha, extrativismo e pesca esportiva. Informações genéticas destas espécies são de fundamental importância para seu manejo e produção. Mesmo assim, das 13.000 espécies de peixes marinhos descritos apenas 2% foram estudadas sob o ponto de vista citogenético e menos de 1% sobre suas características reprodutivas. A reprodução induzida, a citogenética e a criopreservação de gametas, representam importantes áreas aplicadas de estudo em peixes. No presente projeto análises citogenéticas foram empregadas na caracterização genética de espécies da família Gerreidae, ocorrentes no litoral e regiões estuarinas do Nordeste do Brasil. Diferentes métodos de identificação de regiões cromossômicas foram empregados por meio de técnicas convencionais (Ag-RONs, bandamento C), coloração com fluorocromos base-específicos (DAPI-CMA3), e mapeamento cromossômico de genes ribossomais marcadores DNAr 18S e 5S, através da hibridação in situ com sondas fluorescentes (FISH). As seis espécies analisadas revelaram marcante conservadorismo cromossômico. Os genes ribossomais 18S e 5S quando analisados em perspectiva filogenética demonstram dinâmica evolutiva variada, podendo apresentar estase em alguns grupos e maior dinamismo em outros. As análises por duplo-FISH dos sítios 18S e 5S se revelaram eficientes marcadores citotaxonômicos nos cariótipos homogêneos deste grupo de espécies. Os padrões cariotípicos identificados, além dos aspectos evolutivos do cariótipo identificados, são sugestivos de baixa potencial de barreiras pós-zigóticas, instigando pesquisas futuras de prospecção de hibridação interespecífica de valor comercial destas espécies.

8
  • DINALVA BRITO DE QUEIROZ
  • Biolipídeo B2: Desenvolvimento de um veículo estável para reposição hormonal transdérmica nanoestruturada

  • Leader : MARCO ANTONIO BOTELHO SOARES
  • MEMBRES DE LA BANQUE :
  • MARCO ANTONIO BOTELHO SOARES
  • DANILO CALDAS DE QUEIROZ
  • JOSE WALLY MENDONÇA MENEZES
  • PIERRE BASILIO ALMEIDA FECHINE
  • RONALDO DE SOUSA RUELA
  • Data: 5 août 2013


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  • Este estudo teve como objetivo avaliar a segurança e a eficácia de um novo produto destinado a Terapia de Reposição Hormonal transdérmica nanoestruturada (TRHN), com base em uma formulação patenteada sob No. WO2012/009778.A2. Tal formulação foi capaz de restabelecer os níveis séricos de  Estradiol (0,1%) + Estradiol (0,25%), em 122 mulheres menopausadas com idade média de 56,88 (± 6,27). A avaliação faz parte de um estudo prospectivo longitudinal. Parâmetros clínicos, incluindo o grau de satisfação com alívio sintomático, as concentrações séricas de estradiol, peso, pressão arterial, foram comparados entre a o início e o final do tratamento. Os achados mostram que o BIOLIPÍDEO B2 foi eficaz e seguro em restabelecer os níveis séricos hormonais sem efeitos colaterais. A satisfação com tratamento foi de 92%. As concentrações no soro de estradiol foi significativamente maior após tratamento (p <0,05). Peso e pressão arterial sistólica e diastólica não apresentaram diferenças significativas (p> 0,05) ao longo do tratamento. Não Foi observada hemorragia vaginal. Avaliação de mamografia bilateral das mamas o tratamento  encontraram resultados normais em todas as mulheres. Este estudo mostra pela primeira vez a eficácia de uma formulação transdérmica nanoestruturada na entrega transdérmica de estradiol e o estriol medido in vivo utilizando Espectroscopia Raman confocal. A formulação do BIOLIPIDEO B2 é segura e eficaz em restabelecer os níveis séricos de estradiol e de aliviar os sintomas da menopausa. A formulação pode servir como uma boa escolha para a terapia de reposição hormonal para proteger contra sintomas pós-menopausa. 

9
  • GUSTAVO BARBALHO GUEDES EMILIANO
  • Estudo dos efeitos das condições ambientais tropicais ao longo do tempo de exposição sobre o DNA de dentes humanos para genotipagem forense.

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • LELIA BATISTA DE SOUZA
  • LUIZ ANTONIO FERREIRA DA SILVA
  • ROGÉRIO NOGUEIRA DE OLIVEIRA
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • Data: 15 août 2013


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  • As condições ambientais interferem nos processos de decomposição de corpos e remanescentes humanos. Variáveis associadas ao clima e ao solo como alta temperatura, umidade elevada, incidência da radiação solar, composição do solo e o pH do solo podem degradar o DNA de amostras biológicas coletadas de cenas de crimes e de corpos não identificados. Os dentes representam uma importante fonte de DNA, pois são os tecidos mais rígidos do corpo humano e mais resistente à putrefação. Os dentes são coletados quando o corpo encontra-se esqueletizados ou quando os outros tecidos estão em pequenas quantidades e putrefeitos. A genotipagem forense com STRs (Short Tandem Repeat) é um método padrão que apresenta alto poder de discriminação entre indivíduos e a amplificação de amostras degradadas.  A presente pesquisa objetivou avaliar a quantidade e qualidade do DNA de amostras dentárias expostas às condições ambientais de clima e de solo tropicais por T0, T1 (4 meses), T2 (9 meses) e T3 (24 meses). A amostra foi constituída por 65 dentes molares hígidos de indivíduos com média de idade de 24 anos (dp ± 3). Os dentes foram limpos física-quimicamente e pulverizados criogenicamente. O DNA dentário foi extraído através do protocolo de extração AFDIL, quantificados em PCR em tempo real através do kit Quntifiler® Human DNA Quantification e os STRs autossômicos (FGA, D16S539, D7S820, D13S317, D3S1358, Penta E, D18S51, D12S391, TH01, D19S433, CSF1PO, TPOX, vWA, D5S818, D2S1338, SE33 e a amelogenina) amplificados em reações de PCR multiplex. As análises de variância (ANOVA) e o pós-teste de Tukey mostraram haver diferenças significativas (p ≤ 0,05) nas médias de quantidades de DNA entre o grupo controle e grupo experimental no tempos de exposição T1, T2 e T3 e no grupo superfície entre T1 e T3. A quantificação foi possível em 92% (n = 60) das amostras. A diferença na médias de amplificação dos loci de STRs foi significativa (p<0,05) nos grupos superfície, argila e areia ao longo dos tempos T1, T2 e T3. Em 5 amostras do tempo T3 não houve amplificação. Os três maiores marcadores CSF1PO, FGA e Penta E amplificaram em 60% das amostras da superfície, 94% da argila e 87% da areia no tempo T1. Os três menores marcadores D5S818, D19S433 e D3S1358 amplificaram em 94% das amostras da superfície, 94% da argila e 100% da areia no tempo T1. A amelogenina amplificou em 86% das 65 amostras e 47% no grupo experimental no tempo T3. Houve variação significativa (p<0,05) para as médias das áreas da amelogenina entre o grupo controle e o grupo experimental em T1 e T3. Para a área do marcador CSF1PO houve variação significativa (p<0,05) entre o grupo controle e o grupo experimental no tempo T1. Face ao exposto, conclui-se que a submissão dos dentes às condições de clima e de solo foi determinante para a redução significativa da quantidade média de DNA no primeiro tempo de exposição T1 (4 meses) e os STRs podem ser indicados eficientemente para genotipagem com até 4 meses de exposição às condições de clima e de solo tropicais.

2012
Thèses
1
  • FABIO TEIXEIRA DUARTE
  • Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio.

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • JOAO PAULO MATOS SANTOS LIMA
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • THALLES BARBOSA GRANGEIRO
  • VIVIANE SOUZA DO AMARAL
  • Data: 30 mars 2012


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  • Chromobacterium violaceum é um bacilo Gram-negativo, de vida livre, saprófito e patógeno oportunista que habita no solo e na água na Amazônia. Tem como característica principal a produção de um pigmento violeta,  a violaceína.  Esse pigmento possui várias aplicações farmacológicas e biotecnológicas. Devido ao grande  potencial  biotecnológico,  seu  genoma  foi  totalmente  sequenciado  em 2003.  A análise do genoma mostrou que esse organismo possui vários genes relacionados  à  aclimatação  ambiental  e  à  tolerância  ao  estresse.  O  objetivo deste  trabalho  foi  estudar  o  proteoma  diferencial  em shotgun de C. violaceum em  resposta  ao  estresse  oxidativo  provocado  por  H2O.  Para  tal, colônias de C. violaceum foram cultivadas na presença e na ausência de 8 mM de  H2O2 por  duas horas.  Após  este  período as  proteínas  totais  da  bactéria foram  extraídas  utilizando  tampão  solubilização  e  dosadas  por  Bradford. Proteínas em um total de 20 µg foram submetidas a SDS-PAGE e ao final, as proteínas foram fixadas e coradas com comassie blue. Cada coluna do gel foi cortado  em  9 pedaços,  os  quais  foram  cortados  em  tamanhos  menores  para assim serem descorados. As proteínas do gel foram então alquiladas e tiveram as  pontes  de  hidrogênio  reduzidas para,  em  seguida,  serem digeridas  pela tripsina. Os peptídeos trípticos gerados foram submetidos ao espectrômetro de massas tipo Orbitrap.  Pacotes  de  programas  de  bioinformática  auxiliaram  na interpretação  dos  espetros  detectados  e  na  definição  das  proteínas diferencialmente  expressas.  A  proteômica  de  shotgun  revelou  que C violaceum expressou 131  proteínas  exclusivamente  na  condição  controle,  177 proteínas  passaram  a  ser  expressas  sob  estresse  oxidativo  e  1175  proteínas possuem expressão  nas  duas  condições.  Análise  dos  resultados mostrou  que na  condição  de  estresse  oxidativo,  essa  bactéria  muda  sua  fisiologia, expressando  proteínas  capazes  de;  detoxicar  o  H2O,  proteger  biomoléculas, degradar  moléculas  oxidadas,  combater  ciclo  lítico,  modificar  estruturas morfológicas,  modelar  expressão  gênica,  modificar  proteínas  entre  outras. Sugere-se então, que C. violaceum possui mecanismos complexos, eficientes e versáteis  em  resposta  estresse  oxidativo.  Várias  ORFs  hipotéticas foram detectadas nesse estudo e futuramente terão suas funções caracterizadas. 

     

2
  • DEBORAH AFONSO CORNELIO
  • Análise da estabilidade genética de células tronco mesenquimais humanas.

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • ANDRE DUCATI LUCHESSI
  • DANIEL CARLOS FERREIRA LANZA
  • MARCOS ROMUALDO COSTA
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • SILVIA REGINA CAMINADA DE TOLEDO
  • Data: 13 avr. 2012


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  • Células tronco mesenquimais humanas (CTMH) são células multipotentes utilizadas em várias pesquisas de terapia celular regenerativa, pois apresentam a capacidade de se diferenciar em múltiplas e diferentes linhagens, têm grande capacidade de expansão in vitro, excelentes propriedades imunossupressoras e são capazes de secretar moléculas bioativas que exercem efeitos tróficos.  O cordão umbilical é uma fonte de CTMH cuja extração não necessita de um procedimento invasivo, além de não envolver controvérsias éticas, políticas e religiosas. Um dos problemas que pode ocorrer em linhagens de CTMH de diferentes fontes é a possibilidade de ocorrência de alterações cromossômicas e instabilidade genômica, que podem aparecer durante a expansão in vitro. Além disso, as CTMH de um dos cordões apresentaram uma alteração cromossômica constitucional: inversão paracêntrica no braço curto do cromossomo 3, cariótipo: 46,XY,inv(3)(p13p25~26). Em 3p25-26, estão localizados vários genes de grande importância biológica, como genes envolvidos com o reparo de DNA e outros responsáveis pelo desenvolvimento de tumores. O titânio é um dos materiais mais utilizado para fabricação de implantes ortopédicos e dentários, e é considerado um excelente biomaterial, entretanto as partículas derivadas de próteses acumulam-se nos tecidos periprostéticos e na medula óssea local, disseminam-se para linfonodos, fígado e baço. As implicações biológicas a longo prazo da disseminação sistêmica de partículas de metais e seus efeitos cito- e genotóxicos não estão bem caracterizados. Considerando-se que possíveis alterações  em genes em 3p25~26 poderia predispor as CTMH com inv(3) à transformação espontânea in vitro, ainda mais no cultivo na presença de titânio, neste trabalho investigamos a estabilidade genética de CTMH isoladas da veia do cordão umbilical durante a expansão in vitro, após o congelamento, e verificamos as características das CTMH com cariótipo alterado comparadas com CTMH com cariótipo normal em diferentes condições de cultivo in vitro, na presença e na ausência de titânio, antes e após a senescência. A análise da estabilidade genômica das CTMH foi realizada através da análise do cariótipo e da análise do citoma / micronúcleo das CTMH. Ficou estabelecido que as CTMH são capazes de internalizar as partículas de titânio, mas as células mantém sua capacidade de proliferação, diferenciação e preservam os mesmos  marcadores de membrana. Além disso, demonstrou-se que existe um aumento na instabilidade genômica com o decorrer do tempo de expansão in vitro, e esta instabilidade foi maior na presença de grande concentração de partículas de titânio. É necessário sempre avaliar os riscos/ benefícios da utilização do titânio na terapia tecidual envolvendo CTMH, considerando a biossegurança da utilização da regeneração óssea guiada que utiliza CTMH e titânio. Mesmo não se utilizando o titânio, é importante que o uso terapêutico de tais células seja baseado em análises que garantam a qualidade, segurança e estabilidade celular, com a implementação de programas de controle de qualidade adequados. Como conclusão, sugere-se que a análise citogenética  e a análise do citoma sejam realizadas nas CTMH antes de implantar num paciente, sejam elas cultivadas por longo tempo ou não.

3
  • BENEDITA MARTA GOMES COSTA
  • Da bancada ao bureau: análise do desenvolvimento da Biotecnologia no nordeste brasileiro.

  • Leader : GORETE RIBEIRO DE MACEDO
  • MEMBRES DE LA BANQUE :
  • ANA CRISTINA DE ALMEIDA FERNANDES
  • EDILSON DA SILVA PEDRO
  • EDNA MARIA DA SILVA
  • ELZA FERNANDES DE ARAÚJO
  • GORETE RIBEIRO DE MACEDO
  • ZULMARA VIRGINIA DE CARVALHO
  • Data: 17 avr. 2012


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  • O ponto de partida consistiu em investigar o desenvolvimento da Biotecnologia no nordestes brasileiro na perspectiva de um Sistema Regional de Inovação (SRI) em Biotecnologia. Diferentes recortes foram realizados ao longo do estudo, procurando, inicialmente, descrever as características de um sistema de inovação no nível regional. Nessa trajetória, percebemos que o contexto institucional, o fluxo do conhecimento e o financiamento se constituem em elementos essenciais para analisar o desenvolvimento da Biotecnologia na perspectiva de um SIR. Para a análise das dimensões utilizamos a complementaridade de metodologias (quantitativas e qualitativas) a fim de se compreender a dinâmica da Biotecnologia na região nordeste. A análise possibilitou verificar que as políticas de integração na área CT&I desempenham papel essencial para o desenvolvimento da Biotecnologia na região. As atividades desenvolvidas pelas instituições realizam-se praticamente no nível intrainstitucional e inter-regional. O conhecimento gerado no âmbito empresarail (bureau) encontra-se atrelado às áreas de Agropecuária, Saúde e Industrial da PDB, coincidindo com a produção científica da região (bancada). O financiamento a CT&I em Biotecnologia encontra-se dependente das agências de fomento vinculadas à esfera federal, no entanto, essa dinâmica tem proporcionado mudanças no cenário regional.

4
  • EURICO AZEVEDO DIAS JUNIOR
  • Estrutura genética populacional de Cioba e Dentão (Lutjanidae) ao longo do litoral brasileiro - implicações na conservação.


  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • CARLOS ALFREDO GALINDO BLAHA
  • LILIANE DE LIMA GURGEL
  • MAISA CLARI FARIAS BARBALHO DE MENDONÇA
  • VITOR DE OLIVEIRA LUNARDI
  • WAGNER FRANCO MOLINA
  • Data: 1 juin 2012


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  • Espécies   Atlânticas   da   família   Lutjanidae,   conhecidas   como  snappers,   têm   sido amplamente   estudadas   sobre   diversos   aspectos   biológicos,   contudo   a   estruturação genética   de   suas   populações   é  em   grande   parte   desconhecida.  Lutjanus   analis  (mutton snapper)   com   extensa   distribuição   geográfica   no   Atlântico   têm   sido   historicamente sobrexplorada ao  longo da  costa brasileira  mesmo  na  ausência  de  informações  sobre  o status de divergência genética de suas populações. Análises das sequências hipervariáveis HVR 1 do DNA mitocondrial de amostras de extensa área do litoral brasileiro indicam que L. analis,   nesta   região   do   Atlântico,   pode   ser   considerada   como   uma   única   população panmítica com alta diversidade genética, cujos padrões histórico-demográficos demonstram ter sofrido efeitos de eventos ocorridos no Pleistoceno.

5
  • UEDSON PEREIRA JACOBINA
  • PARÂMETROS CITOGENÉTICOS DE ESPÉCIES COMERCIAIS DE CARANGIDAE (PERCIFORMES), COM VISTAS A SUA EMPREGABILIDADE NA CONSERVAÇÃO BIOLÓGICA E PISCICULTURA MARINHA.

  • Leader : WAGNER FRANCO MOLINA
  • MEMBRES DE LA BANQUE :
  • WAGNER FRANCO MOLINA
  • CARLOS ALFREDO GALINDO BLAHA
  • LILIANE DE LIMA GURGEL
  • ORLANDO MOREIRA FILHO
  • VITOR DE OLIVEIRA LUNARDI
  • Data: 28 sept. 2012


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  • Entre os peixes marinhos, as famílias Carangidae e  Rachycentridae se apresentam como grupos de grande importância comercial pela pesca e potencial para piscicultura marinha. Entretanto, bases genéticas que possam alicerçar o futuro cultivo destas espécies, sobretudo seus aspectos citogenéticos são incipientes. Os padrões cromossômicos têm fornecidodados básicos para a prospecção de processos biotecnológicos de manipulação cromossômica voltados ao melhoramento genético, como a indução a poliploidia, ginogênese e androgênese, assim como obtenção de estoques monossexo e hibridizações interespecíficas. Neste trabalho é apresentado um amplo levantamento citogenético em 10 espécies, sendo sete da família  Carangidae e de Rachycentron canadum, espécie monotípica da família Rachycentridae. A caracterização citogenética clássica e mapeamento  in situde sequências multigênicas foram empregadas. Adicionalmente em espécies do gênero  Selene  e em morfótipos de Caranx lugubrisforam realizadas comparações através de morfometria geométrica. Em geral, as espécies exibiram um marcante conservadorismo cromossômico numérico (2n=48). Apesar de apresentar, em grande parte, fórmulas cariotípicas diferenciadas, conservam diversas características cromossômicas típicas da Ordem Perciformes, como elevado número de elementos monobraquiais, sítios Ag-RONs/ DNAr 18S simples e heterocromatina reduzida, preferencialmente centromérica. Os principais mecanismos envolvidos na diversificação cariotípica são as inversões pericêntricas, com ação secundária de fusões cêntricas. Além do mapeamento físico e detalhamento cromossômico para as espéciessão apresentados e discutidos padrões de variabilidade intra e diversificação interespecíficas, com identificação de marcadores citotaxonômicos. Este conjunto dos dados propicia um melhor conhecimento dos padrões carioevolutivos destes grupos e condições para o desenvolvimento de protocolos biotecnológicos baseados na manipulação cromossômica para estas espécies Atlânticas.


6
  • GABRIELA VASCONCELOS DE ANDRADE
  • CARACTERIZAÇÃO HEMATOLÓGICA E IMUNOFENOTÍPICA EM 192 PACIENTES COM LEUCEMIA LINFÓIDE AGUDA.

  • Leader : GERALDO BARROSO CAVALCANTI JUNIOR
  • MEMBRES DE LA BANQUE :
  • GERALDO BARROSO CAVALCANTI JUNIOR
  • IVONETE BATISTA DE ARAUJO
  • MARIA GORETTI DO NASCIMENTO SANTOS
  • AMALIA CINTHIA MENESES DO REGO
  • DEBORAH AFONSO CORNELIO
  • Data: 1 nov. 2012


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  • A Leucemia Linfóide Aguda (LLA) é uma doença de caráter maligno do sistema  imune  que  se  origina  a  partir  da  proliferação  neoplásica  de  uma  única  célula  linfocitária  progenitora  que  sofre  alteração  no  seu  processo  de  crescimento  e  reprodução  resultando  na  formação  de  um  clone  neoplásico  (teoria  monoclonal).  As  células  deste  clone  vão  se  expandindo  ao nível  dos  diversos órgãos linfóides e da medula óssea, atingindo a corrente sanguínea, podendo assim  atingir  vários  órgãos  ou  tecidos.  Atualmente  as  LLAs  são  classificadas pelo aspecto morfológico associado às características do perfil imunológico das células  malignas.  O  grupo  French-American-British  (FAB)  classificou  as  LLAs em  três  subtipos  distintos  baseados  somente  em  critérios  morfológicos  de medula  óssea  e  de  sangue  periférico  em  L1,  L2  e  L3.  Métodos  de imunofenotipagens  por  citometria  de  fluxo  tornaram  possível  a  detecção  de populações de células específicas obtidas de suspensões celulares através do uso  dos  anticorpos  monoclonais.  A  proposta  deste  estudo  foi  diagnosticar  e classificar  192  pacientes  com  LLA  com  base  na  imunofenotipagem  por citometria de fluxo com um painel de AcMo específico para leucemias agudas. Foram realizadas análises e correlações entre os dados demográficos, clínicos, laboratoriais  e  imunofenotípicos  desses  pacientes.  A  expressão  de  antígenos linfóides  como  o  CD3  e  CD19,  para  as  LLAs  T  e  B,  respectivamente,  associados na maioria dos casos ao CD34 caracterizou imunofenotipicamente  a  presença  de  precursores  linfóides  muito  imaturos.  A  expressão  dos marcadores linfóides B mais comuns nos pacientes com LLA foram os CD19 e sCD22,  e  dos  marcadores  linfóides  T  foram  os  CD7  e  cCD3.  Foi  notificada  a expressão  de  fenótipos  aberrantes  quando  observamos  a  expressão  dos marcadores linfóides T mais comuns nos pacientes com LLA B como o sCD3, e também  quando  observamos  a  expressão  dos  marcadores  linfóides  B  mais comuns nos pacientes com LLA T como os CD19 e o cCD79a. Concluímos que a imunofenotipagem por citometria de fluxo é uma metodologia necessária para o diagnóstico e a classificação das leucemias agudas.

2011
Thèses
1
  • LOURENA MAFRA VERISSIMO
  • Desenvolvimento de Nanosistemas Farmacêuticos para Terapia Gênica.

  • Leader : ERYVALDO SOCRATES TABOSA DO EGITO
  • MEMBRES DE LA BANQUE :
  • ELIAS FATTAL
  • ERYVALDO SOCRATES TABOSA DO EGITO
  • HELDER FERREIRA TEIXEIRA
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • MATHEUS DE FREITAS FERNANDES PEDROSA
  • SELMA MARIA BEZERRA JERONIMO
  • Data: 17 mars 2011


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  • A terapia gênica é um dos maiores desafios propostos pela pesquisa pós-genômica e se baseia na transferência de material genético a uma célula, tecido ou órgão com o intuito de curar ou melhorar o estado clínico do paciente. Em sua forma mais simples, a terapia gênica consiste na inserção de genes funcionais em células com genes defeituosos objetivando substituir, complementar ou inibir esses genes causadores de doenças. Para que o DNA exógeno seja expresso em uma população celular faz-se necessária a sua transferência até o local de ação. Assim, é necessário criar veículos, os vetores, que transportem e protejam o DNA até que este chegue a uma população celular alvo. Os obstáculos encontrados com a utilização de vetores virais têm proporcionado o interesse no desenvolvimento de vetores não-virais, por serem fáceis de produzir, apresentarem estabilidade controlável e facilitarem a transfecção gênica. O objetivo deste trabalho foi avaliar dois diferentes vetores não virais, lipossomas e nanoemulsões catiônicos, e sua possível utilização na Terapia Gênica. Para isso, utilizaram-se lipídeos catiônicos e co-tensoativos na produção dos dois sistemas. As nanoemulsões foram produzidas pelo método de sonicação e compostas por Captex® 355; Tween® 80; Spam® 80; lipídeo catiônico, Estearilamina (EA) ou N-[1-(2,3-Dioleoiloxi)propil]-N,N,Ntrimetilamonio metilsulfato (DOTAP); e água ultra-pura (Milli-Q® ). Estes sistemas foram caracterizados analisando-se tamanho médio de gotícula, índice de polidispersão (PI) e potencial zeta. Avaliou-se ainda a estabilidade dos sistemas e suas capacidades de compactação do material genético. Os lipossomas foram preparados a partir do método de hidratação do filme e compostos por DOTAP, Dioleilfosfatidiletanolamina (DOPE), na presença ou ausência de Rodaminafosfatidiletanolamine (PE-Rodamina) e do conjugado Ácido Hialurônico –DOPE (HA-DOPE). Estes sistemas foram caracterizados da mesma forma das nanoemulsões e também se avaliaram estabilidade; influência do tempo, tamanho de material genético e presença ou ausência de endotoxinas na formação dos lipoplexos. Os resultados obtidos permitem afirmar que os sistemas são promissores para posterior utilização na terapia gênica e que esta área promete ser uma área fértil de pesquisa científica e clínica por muitos anos, e provavelmente se tornará uma prática clínica importante neste século. No entanto, da possibilidade à prática existe um longo caminho a percorrer.

2
  • DELANNE CRISTINA SOUZA DE SENA FONTINELE
  • Identificação de genes em Chromobacterium violaceum Relacionados à Resposta ao Estresse.

  • Leader : LUCYMARA FASSARELLA AGNEZ LIMA
  • MEMBRES DE LA BANQUE :
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • HUGO ALEXANDRE DE OLIVEIRA ROCHA
  • ADRIANA FERREIRA UCHOA
  • KERONNINN MORENO DE LIMA BESSA
  • TIRZAH BRAZ PETTA
  • Data: 8 sept. 2011


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  • O seqüenciamento do genoma da espécie Chromobacterium violaceum identificou um cromossomo simples circular de 4.8 Kb, onde aproximadamente 40% das ORFs encontradas são classificadas como hipotéticas conservadas ou hipotéticas. Algumas regiões gênicas de interesse biotecnológico vêm sendo caracterizadas, como por exemplo, genes de reparo de DNA. Diante disso, objetivou-se identificar genes de  C. violaceum envolvidos com mecanismos de reparo de DNA e manutenção da integridade genômica. Foram realizados ensaios de complementação funcional com uma biblioteca genômica de  C. violaceum transformada na cepa DH10B de Escherichia coli deficiente em RecA. Cinco clones foram selecionados após ensaios com luz UVC. No clone PLH6A foram identificadas quatro ORFs (CV_3721 a 3724). Duas ORFs, CV_3722 e  CV_3724, foram subclonadas e uma atividade sinérgica de complementação foi observada. O ensaio de mutagênese com rifampicina após tratamento de luz UVC mostrou alta freqüência de mutagenicidade para a ORF CV_3722 (Pol III δ’ subunit). Dessa forma, propomos que a subunidade  δ’ de C. violaceum pode agir em DH10B na síntese translesão utilizando Pol IV em uma via  RecA independente. A ocorrência de um operon foi
    confirmada usando cDNA de  C. violaceum em um ensaio de RT-PCR. Adicionalmente, foi observada a indução do operon após tratamento com UVC, dessa forma, esse operon composto pelas quatro ORFs foi identificado estar associado a resposta ao estresse em  C. violaceum. Em ensaios de curva de crescimento os outros quatro clones foram capazes de complementar a função na dose de 5 J/m2. E em ensaios de mutagenicidade três deles apresentaram freqüências de mutação com diferenças significativas em relação ao controle (DH10B), demonstrando que de alguma forma eles estão envolvidos com a resposta ao estresse em C. violaceum e manutenção da integridade genômica. Os resultados obtidos contribuem para um melhor conhecimento da genética desta espécie. 

3
  • VIVIANE KATIELLY SILVA MEDEIROS
  • Análise proteômica de Chromobacterium violaceum: expressão basal e diferencial após exposição à luz UVC.

  • Leader : SILVIA REGINA BATISTUZZO DE MEDEIROS
  • MEMBRES DE LA BANQUE :
  • SILVIA REGINA BATISTUZZO DE MEDEIROS
  • LUCYMARA FASSARELLA AGNEZ LIMA
  • JOAO PAULO MATOS SANTOS LIMA
  • CARLOS PRIMINHO PIROVANI
  • THALLES BARBOSA GRANGEIRO
  • Data: 13 déc. 2011


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  • Chromobacterium  violaceum  é  um  bacilo  de  vida-livre,  Gram-negativo comumente encontrado no solo e nas águas de regiões tropicais e subtropicais. Uma  das  principais  características  deste  organismo  é  sua  capacidade  de produzir  o  pigmento  violaceína,  o  qual  apresenta  inúmeras  atividades biológicas.  Em  2003,  o  genoma  deste  organismo  foi  completamente  sequenciado  e  revelou  informações  importantes  sobre  a  fisiologia  desta bactéria.  Porém,  poucos  estudos  pós-genômicos  tem  sido  realizados.  Este trabalho avaliou o perfil proteico de C. violaceum cultivada em meio LB a 28ºC, o que permitiu a identificação de proteínas relacionadas a um possível sistema de  secreção  ainda  não  identificado  e  caracterizado  em  C.  violaceum,  ao sistema  quorum sensing,  a  processos  regulatórios  da  transcrição  e  tradução, adaptação  ao  estresse  e  ao  potencial  biotecnológico.  Além  disso,  a  resposta desta  bactéria  à  radiação  UVC  foi  avaliada.  A  comparação  do  perfil  protéico, analisado  por  eletroforese  2-D,  do  controle  versus  tratado  possibilitou  a identificação  de  52  proteínas  que  surgiram  após  a  indução  do  estresse.  Os resultados  obtidos  permitiram  a  elaboração  de  uma  via  de  resposta  de  C. violaceum  ao  estresse  gerado  pela  luz  UVC.  Esta  via,  que  parece  ser  de resposta  geral  ao  estresse,  envolve  a  expressão  de  proteínas  relacionada  à divisão  celular,  metabolismo  de  purinas  e  pirimidinas,  choque  térmico  ou chaperonas,  fornecimento  de  energia,  regulação  da  formação  de  biofilme, transporte,  regulação  do  ciclo  lítico  de  bacteriófagos,  além  de  proteínas  que ainda  não  apresentam  função  caracterizada.  Apesar  da  reposta  apresentar similaridades com a SOS clássica de E. coli, ainda não podemos afirmar que C. violaceum apresenta uma resposta SOS-like, principalmente devido a ausência da caracterização de um proteína LexA-like neste organismo.

2010
Thèses
1
  • CRISTIANE FERNANDES DE ASSIS
  • Produção e Caracterização de Quitooligossacarídeos produzidos por fungos e avaliação da citotoxicidade em células tumorais.

  • Leader : EVERALDO SILVINO DOS SANTOS
  • MEMBRES DE LA BANQUE :
  • ELIANA SETSUKO KAMIMURA
  • EVERALDO SILVINO DOS SANTOS
  • LUCIANA ROCHA BARROS GONÇALVES
  • MARIA DE FATIMA VITORIA DE MOURA
  • SUELI RODRIGUES
  • Data: 18 janv. 2010


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  • A quitosana é um polímero natural, biodegradável, não tóxico e de alta massa molecular
    obtido  a  partir  de  animais  marinhos,  insetos  e  microrganismos.  Os  oligômeros  de
    glicosamina  (GlcN)  e  N-acetilglicosamina  (GlcNAc)  têm  atividades  biológicas
    interessantes,  incluindo  efeitos  antitumorais,  atividade  antimicrobiana,  antioxidante
    entre  outras.  A  alternativa  proposta  por  este  trabalho  foi  estudar  a  viabilidade  de
    produção de quitooligossacarídeos utilizando um  extrato bruto de  enzimas produzidas
    pelo fungo Metarhizium anisopliae. A hidrólise da quitosana foi realizada em diferentes
    tempos,  de  10  a  60 minutos  para  a  produção  de  quitooligossacarídeos  e  a  detecção  e
    quantificação foi realizado pela Cromatografia Liquída de Alta Eficiência. A avaliação
    da  citotoxicidade  dos  oligômeros  de  quitosana  foi  realizada  em  células  tumorais
    (HepG2 e HeLa) e não  tumoral (3T3). As células foram  tratadas durante 72 horas com
    os oligômeros a viabilidade celular foi feita usando o método do MTT. A produção de
    oligômeros de quitosana  teve maiores rendimentos durante 10 minutos de hidrólise, os
    pentâmeros  apresentaram  concentração  0,15  mg/mL,  porém  os  hexâmeros,  que
    apresentam maior interesse pelas suas propriedades biológicas, só foram detectados com
    30 minutos de hidrólise  apresentando uma  concentração de 0,004 mg/mL. Um estudo comparando  os  oligômeros  puros  de  quitosana  com  a  mistura  dos  oligômeros
    produzidos  pelo  extrato  bruto  enzimático  na  toxicidade  das  células  tumorais  revelou
    maior  eficiência  da mistura  dos  quitooligossacarídeos. Além  disso,  o  composto  com
    maior citotoxicidade entre os oligômeros puros foi a glicosamina, apresentando valores
    de IC50 0.30; 0.49; 0.44 mg/mL para células HepG2, HeLa e 3T3, respectivamente. Os
    oligômeros  produzidos  por  hidrólise  durante  20  minutos  foram  analisados  quanto  à
    capacidade  de  inibir  células  tumorais mostrando  inibição  da  proliferação  apenas  nas células  HeLa,  não  apresentando  nenhum  efeito  em  células  HepG2  e  células  de
    fibroblastos (3T3). 

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