DSpace Coleção:
http://repositorio.ufc.br/handle/riufc/15581
2024-03-28T08:27:15ZProdução da uma galectina recombinante de anuros a partir da bioprospecção e mapeamento tecnológico das galectinas-9
http://repositorio.ufc.br/handle/riufc/76195
Título: Produção da uma galectina recombinante de anuros a partir da bioprospecção e mapeamento tecnológico das galectinas-9
Autor(es): Rodrigues, Cássia Ferreira
Abstract: Galectins are a family of animal lectins that have the ability to bind reversibly to carbohydrates with a high affinity for β-galactosides. They are expressed in different organisms and are involved in cell adhesion, tumor differentiation, apoptosis and have thus been the subject of technological product development. As few amphibian galectins have a primary structure or a solved three-dimensional structure, the structural elucidation of these lectins will make it possible to reveal their mechanisms of action within cells and evaluate their biotechnological potential. Thus, the aim of this work was to produce a recombinant galectin encoded in the anuran genome based on information obtained from molecular bioprospecting and a review of patents related to galectins. The galectin present in the secretion of the paratoid gland of Rhinella diptycha was partially purified using ion exchange chromatography and the fractions obtained from the chromatography were subjected to the hemagglutination test with rabbit erythrocytes and it was possible to identify the presence of the active galectin in one of the fractions. Using mass spectrometry, it was possible to obtain the primary structure of R. diptycha galectin, which is composed of 393 amino acid residues with a molecular mass of 51 kDa and a triple domain. Through a search for protein sequences similar to R. diptycha galectin in the National Center for Biotechnology Information (NCBI) database, B. bufo galectin-9 was found to be highly similar to R. diptycha galectin, which is why it was decided to produce it heterologously. Thus, two strains of Komagataella pastoris were transformed with the expression vector pPICZα(A) containing the gene encoding galectin, after codon optimization. Subsequently, the target protein was expressed through induction with methanol, and a molecule with a mass of 48 kDa was obtained, referring to recombinant galectin-9. Using the data available in the European Patent Office (EPO) and Cortellis Drug Discovery Intelligence (CDDI) databases, technological advances related to galectin-9 were mapped. In this survey, galectin-9 is related to various diseases, including several types of cancer, and has been used as a target for treatment or as a biomarker for cancer diagnosis. Therefore, galectin-9 with a triple secretion domain from R. diptycha, and recombinant galectin-9 from B. bufo are proteins from a family with demonstrated efficacy in technological prospecting, in terms of their anticancer activity, and will thus contribute to diagnosis or treatment in response to global demand.
Tipo: Tese2023-01-01T00:00:00ZPerfil antropométrico e análise proteômica do plasma seminal: estratégia para identificação de marcadores moleculares da fertilidade em homens atendidos em um centro de referência em medicina reprodutiva em Fortaleza, Ceará
http://repositorio.ufc.br/handle/riufc/74863
Título: Perfil antropométrico e análise proteômica do plasma seminal: estratégia para identificação de marcadores moleculares da fertilidade em homens atendidos em um centro de referência em medicina reprodutiva em Fortaleza, Ceará
Autor(es): Cunha, Luana Mara Silva de Castro Pacheco da
Abstract: Male infertility represents a public health problem with negative repercussions on male health, with central obesity and varicocele being risk factors that lead to significant seminal changes, making pregnancy impossible. In this sense, this thesis had two objectives: 1) to evaluate the association between anthropometric markers (BMI, WC) and clinical parameters, including the lifestyle of men undergoing reproductive treatment and 2) to identify biomarkers for male infertility based on the characterization Proteomics of seminal plasma from men attended at an assisted reproduction clinic. The study included 117 men attended at the reproductive medicine center from October 2019 to March 2020. Weight (kg) and waist circumference (cm) were measured and a socioeconomic and lifestyle form was applied. Semen collection was performed to obtain the seminal report and proteomic analysis. In research 1, 98 participants were divided into three groups and categorized by BMI, namely: eutrophic (n = 14), overweight (n = 56) and obese (n = 28). The mean age was 37.8 (6.8) years and the WC was 96.3 (15.3) cm. There was a significant association between BMI and WC (p<0.001), BMI and family diseases (p=0.017), highlighting obesity (p=0.017) and diabetes (p=0.008), as well as medication use (p=0.049). For research 2, we focused on the proteomic analysis where 19 men with a mean age of 35.9 (6.4) years were divided into 4 groups, normozoospermic (NZ) (n=5), varicocele and asthenozospermia (VAZ) (n= 4), varicocele and teratozoospermia (VTZ) (n=4) and, varicocele and oligoteratozoospermia (VOTZ) (n=6). 4,245 proteins were identified in seminal plasma, 724 of them in the NZ group, with emphasis on the most abundant ANX A3, PRDX1 and CRAC1. Then, the VAZ group with 886 proteins with emphasis on IGLC3, IGLC1 and H2BC1, the VOTZ group with 907 proteins, with emphasis on TUBB, MSLN and HEX and the VTZ group, with 1728 proteins, in which HSPA1, KIF5C and CLTCL1, identified as the most abundant. This study provided the association of BMI and familial diseases and medication use, as well as the first characterization of the seminal plasma proteome of men with normal seminal parameters and varicocele, in which it was possible to identify proteins and pathways associated with spermatogenesis, energy metabolism, fertilization, permeability of the reproductive tract and immunofertility.
Keywords: male infertility, obesity, circumference waist, seminal plasma, varicocele, proteome.
Tipo: Tese2023-01-01T00:00:00ZAtividade antibacteriana e antibiofilme de um complexo de rutênio polipiridínico e seu efeito individual e combinado a antibióticos sobre bactérias de interesse clínico
http://repositorio.ufc.br/handle/riufc/74821
Título: Atividade antibacteriana e antibiofilme de um complexo de rutênio polipiridínico e seu efeito individual e combinado a antibióticos sobre bactérias de interesse clínico
Autor(es): Sousa, Leonardo Silva de
Abstract: Healthcare-associated infections (HAIs) are one of the main sources of preventable diseases in hospitalized patients, and cause significant damage to healthcare resources. They are often caused by bacteria resistant to several antibiotics such as methicillin-resistant Staphylococcus aureus – MRSA and long periods of hospitalization. The objective of this study was to evaluate the antibacterial and antibiofilm activity of a polypyridine ruthenium complex designated ruthenium(II) tris(4-methyl-4-carboxamide(2-anthracenyl)-2,2-bipyridine) [Ru(Ant)]32+ on bacterial strains S. aureus (ATCC 25923 and ATCC 700698 MRSA), and S. epidermidis (ATCC 12228 and ATCC 35984 MRSA) of clinical interest. To determine antibacterial activity, compounds were diluted to concentrations ranging from 3.9 to 250 μg/mL. The ruthenium complex [Ru(Ant)]32+ was evaluated for minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in relation to S. aureus ATCC 25923, S. aureus ATCC 700698, S. epidermidis ATCC 12228 and S. epidermidis ATCC 35984, through microdilution tests. The determined MICs were combined with the antibiotics ampicillin and tetracycline, and their effects were evaluated using the checkerboard technique. Antibiofilm activity was evaluated by quantifying biomass using crystal violet staining (CV) and the number of viable biofilm cells using colony forming units (CFU) and measuring metabolic activity using the XTT reduction assay. The effect of the treatment was also analyzed using scanning electron microscopy – SEM and confocal laser microscopy using the Live/Dead cell viability kit. In addition, the cytotoxicity of the complex was evaluated in in vitro tests on murine fibroblast lines. The results obtained showed an MIC value of 31.25 μg/mL for the strains S. aureus ATCC 700698 and S. epidermidis ATCC 35984 and an MIC of 125 μg/mL for S. aureus ATCC 25923 and S. epidermidis ATCC 12228 (MRSE). The combination of the complex with the antibiotics ampicillin and tetracycline showed a synergistic and additive effect, depending on the bacterial strain tested. The complex was also able to reduce the formation of biofilms by between 10 and 99%, with a reduction in the number of viable biofilm cells by up to 99.99% and complete inhibition of metabolism in some strains at concentrations ranging from 3.9 to 250.0 μg/mL. In relation to pre-formed biofilms, a reduction of biofilms of up to 80% was observed, and of viable cells by up to 90%, being capable of inhibiting metabolism by up to 80% at a concentration of 250 μg/mL. In relation to SEM, ruptures in the cell wall were observed in all strains treated with CIM. In confocal laser microscopy, the results on S. aureus ATCC 700698 and S. epidermidis ATCC 35984 treated with CIM demonstrated a reduction in cells and biofilm. Regarding cytotoxicity against the L929 strain, cytotoxicity above 125 μg/mL was observed within 24 hours. The results showed that [Ru(Ant)]32+ has potential as an antimicrobial agent or can even assist in the synthesis of new antibacterial agents of clinical interest for the pharmaceutical industry and research institutions.
Tipo: Tese2023-01-01T00:00:00ZInovação biotecnológica para biorremediação de manguezais contaminados com derivados do petróleo
http://repositorio.ufc.br/handle/riufc/74627
Título: Inovação biotecnológica para biorremediação de manguezais contaminados com derivados do petróleo
Autor(es): Alves, Bella Giselly Torres
Abstract: Mangroves are highly vulnerable to oil spills. The decontamination of these environments by hydrocarbon-degrading microorganisms stands out as a sustainable bioremediation alternative. This study aimed to validate a consortium of two hydrocarbonoclastic actinobacteria, previously isolated from mangrove, identified as Gordonia sp. HEX 5 and Micrococcus sp. HEX 6 for application in bioremediation of hydrocarbon-contaminated soils. First, the presence of the alkB gene in the consortium bacteria was evaluated through PCR using specific primers. Their ability to use hexadecane as a carbon source in mineral medium cultivation was also evaluated. Subsequently, the consortium was immobilized in chitosan spheres and tested in microcosms of mangrove sediment contaminated with 1% hexadecane. Other bioremediation strategies were also carried out in microcosms for comparison and control. The experiment lasted for 60 days, and the tested microcosms received the contaminant at the beginning and after 30 days. Treatments and controls were monitored by measuring dehydrogenase activity (DHA) and changes in bacterial community through sequencing of the 16S ribosomal RNA gene. The results showed that although both bacterial strains used hexadecane as the only carbon source, the alkB gene was detected in only one of them. In the microcosms, evidence of biodegradation was detected on the 3rd day after contamination in the groups inoculated with the consortium, with maximum degradation on the 12th day. After the 2nd contamination, in 30 days, the response was faster and more lasting. Chitosan stimulated the biodegradation, whereas the addition of biosurfactants did not show influence. Significant changes in the bacteriome were observed with the temporal succession of groups involved in biodegradation. At the peak of biodegradation, the bacteriome was enriched with the phyla Proteobacteria, Firmicutes, and Bacteroidetes, with higher abundance of Fusibacter, Flavobacterium, Vibrio, and Fontimonas. The tested technology proved to be capable of stimulating the autochthonous microbiota and accelerating alkane degradation in microcosms.This study gave rise to a product called BioClineX, with a patent submitted to INPI. It also gave rise to the startup Biotech4life Solutions, enabling the transformation of knowledge derived from science into marketable products that have a positive impact on the market and society.
Tipo: Tese2023-01-01T00:00:00Z