https://repositorio.ufpb.br/jspui/handle/tede/338 PGBIOTECM 2026-04-11T14:09:05Z https://repositorio.ufpb.br/jspui/handle/123456789/37754 Título: Potencial antidiabético do ácido ferúlico em modelo experimental - uma revisão integrativa Autor(es): Sousa, Monike de Orientador: Vieira, Giciane Carvalho Abstract: Diabetes mellitus is a metabolic disorder characterized by hyperglycemia from defects in insulin secretion, insulin action or both. The increased amount of glucose in the blood causes energy deprivation for the cells. Type 1 diabetes mellitus and type 2 diabetes mellitus are the most common types of diabetes, however, there is gestational diabetes mellitus and two other specific types of diabetes. This disease is a global epidemic and its numbers increase exponentially each year. In addition to hyperglycemia, diabetes causes several complications affecting almost all tissues and over time can lead to serious complications. Ferulic acid is a phenolic acid found in several vegetables and known for its antioxidant properties, anti-inflammatory and antidiabetic potential. The present work was carried out through a literature review on the most recent results of the use of ferulic acid in the treatment of diabetes. Therefore, a search and survey of publications involving diabetes and the use of ferulic acid was carried out in the SCIELO databases - Scientific Electronic Library online, PubMed and BVS - Virtual Health Library. For compose the sample of selected articles, the following inclusion factors were applied: articles in Portuguese, English or Spanish; that remaining in full and portraying the use of ferulic acid as a potential antidiabetic in an experimental model with rats. The exclusion factors: articles that do not assess the hypoglycemic activity of the substance; research published and indexed outside the period from 2016 to 2021. The sample consisted of a total of 6 articles. Studies have shown possible ways to explain the antidiabetic mechanism of ferulic acid, including the inhibitory activity of peroxiredoxin 1, its effect against cellular stress and action in diabetic neuropathy. Ferulic acid is used not only for hyperglycemic control, but also to improve and prevent some complications of diabetes. Therefore, ferulic acid proves to be a promising substance in the study of the treatment of diabetes mellitus. The studies show possible pathways for the antidiabetic mechanism of ferulic acid, including its inhibitory activity against peroxiredoxin 1, its effect against cellular stress and how it also acted in diabetic neuropathy. Through this research we noticed that ferulic acid is used not only for hyperglycemic control, but also to improve and prevent some complications of diabetes. This demonstrates a wide range of options for studying this promissory substance. Editor: Universidade Federal da Paraíba Tipo: Dissertação 2022-09-28T00:00:00Z https://repositorio.ufpb.br/jspui/handle/123456789/37753 Título: Produção e caracterização bioquímica de proteases de fungos isolados de amostras de solos do semiárido pernambucano Autor(es): Freires, Ariane Susan Santos Orientador: Amaral, Ian Porto Gurgel do Abstract: Soils are habitat for a diverse range of macro and microorganisms that form a network of interrelationships capable of allowing and ensuring the maintenance of life in this place. Even with abiotic (climate, temperature, availability of nutrients and water) and biotic factors that interfere with the development of species, fungi are one of the groups of microorganisms most adaptable to environments like this, since these factors end up conferring resistance to these organisms, that use strategies such as the production of secondary metabolites to ensure their maintenance, an example of this are enzymes. Therefore, this work aimed to evaluate the ability of protease production by filamentous fungi (Aspergillus sp., Paecilomyces sp., Penicillium sp. and Cladosporium sp.), isolated from soil samples from the semi-arid region of Pernambuco, and to characterize the enzymatic extract with better relationship between enzymatic index (IE) and fungal biomass. Six isolates were obtained from the microbiological collection at LAPEEMI, and were submitted to macroscopic analysis, microscopic analysis, and their potential for proteolytic production in solid Milk-Agar medium. The production capacity was tested through three processes, being production in milk-agar medium (solid), which served as an initial screening for better peptidases producers. Only four isolates with better EI and tested under submerged production, in half fiber skimmed milk, with agitation of 150 rpm, 37°C and submerged production without agitation, in triplicate in both cultures, for 6 days, after this time the samples were centrifuged to obtain the enzymatic extract. As a result, it is observed that there is a significant difference between the cultivation method with and without agitation, for biomass production and enzymatic production respectively for Paecilomyces sp. (0.368 and 0.602g and 80.3 and 302 U/mL), Penicillium sp. (1.756 and 2.913g; 75.7 and 177 U/mL), Aspergillus sp. (2.852 and 3.759; 91 and 292.7 U/ml). The latter produced more biomass and the same enzymatic level as Paecilomyces sp. which had lower production of biomass in stationary fermentation. So this isolate was selected for the biochemical characterization tests of the enzymes. The enzymatic extract of Paecilomyces sp. was tested for peptidases classes, optimal temperature, thermostability and pH-optimal. Inhibitors for four classes of proteases were used: serine protease, aspartate protease, metalloprotease, cysteine proteases, of which it was observed that only 20.3% of the proteases were inhibited, 10% of which were serine protease, and 10.3% in aspartate protease, demonstrating that most of the enzymes present do not belong to these classes. The enzymatic activity was analyzed at different temperatures (27 to 97 °C) with an increment of 5°C at each new temperature, demonstrating greater activity at 47°C. As for thermostability, proteases from the crude extract, after being incubated at temperatures (27°C; 37°C; 47°C to 97°C), also with increments of 5°C, demonstrated greater stability in the temperature ranges between 27 °C and 47°C. The enzymatic extract showed two peaks of proteolytic activity at different pH, one at alkaline pH (pH 8.0) and the second peak, this higher one, at acidic pH (pH 6.0). In view of the results obtained, it is concluded that the fungi studied were capable of producing proteases by submerged fermentation in both methodologies using skimmed milk as substrate, with emphasis on stationary cultivation, that fiber skimmed milk medium can be considered a good alternative medium for production of these enzymes and that the extract of proteases from Paecilomyces sp. presents desired characteristics for industrial proteases. Editor: Universidade Federal da Paraíba Tipo: Dissertação 2022-09-27T00:00:00Z https://repositorio.ufpb.br/jspui/handle/123456789/37721 Título: Derivado do carvacrol com propriedades antitumorais via modulação de canais de sódio dependentes de voltagem Autor(es): Vasconcelos, Aliny Pereira de Orientador: Araújo, Demetrius Antonio Machado de Abstract: In cancer pathology, ion channels play an important role in the regulation of cell proliferation, tumor initiation, and progression. Cancers with high expression levels of functional voltage-gated ion channels tend to have a higher metastatic capacity, like neuroblastomas. Natural products, plant derivatives, have stood out in the field of medicinal chemistry because of their intrinsic medicinal properties. An example is carvacrol which, in addition to modular ion channels, has antitumor activity. The natural product carvacrol was used as a base for the drug design of the semi-synthetic carvacrol derivatives compounds. This study proposed to investigate the anticancer potential of carvacrol derivatives, which has previously shown an inhibition effect on voltage-gated sodium channels subtype 1.7 (Nav1.7). The effect of the twelve compounds Derivatives of Carvacrol (DC) was evaluated through the Patch Clamp technique in which Nav1.7 currents (INav1.7) were obtained in CHO-hNav1.7 cells. Derivative of Carvacrol 6 (DC6) with better effect, presenting IC50 of 39.26 μM for blocking Nav1.7. The toxicity of DC6 was evaluated in nauplii of Artemia Salinas, and the behavioral toxicological evaluation was performed in Swiss mice using the Irwin test, the compound has shown low toxicity in both experimental models. The cytotoxic effect of DC6 was evaluated by the MTT reduction method in the neuroblastoma cell lines, SH-SY5Y, Neuro-2A, IMR-32M, and SH-EP, presenting IC50 values for the 24 h times of 23.36 μM, 26.29 μM, 0.35 μM, and 50 μM respectively. In normal cell lines, HUVEC and L929 the IC50 of the compound were higher than 100 μM in both cell types. In sub-toxic concentrations, DC6 presented antiproliferative activity in neuroblastomas. Cell migration assay was performed to verify the effect of DC6 on SH-SY5Y cell motility, in which the cell migration rate was reduced within 48 h. To understand the correlation between Nav1.7 blockade and the reduction of cell migration presented by DC6, migration in CHO-hNav1.7 cells was evaluated in the presence of DC6 and 300 nM of tetrodotoxin (TTX). The cell migration rate after 72h of incubation with 300 nM of TTX was 42.79% and for 8 μM of DC6, it was 34.59%. Suggesting that the decrease in cell migration observed with DC6 treatment occurs mainly via Nav1.7 blockade. To evaluate the mechanism of cell death, the induction of apoptosis test by flow cytometry in neuroblastomas was performed, in which DC6 promoted the induction of cell death in a concentration-dependent manner, via late apoptosis and necrosis. Morphological analysis by fluorescence microscopy was performed on the SH-SY5Y cell line, and after 24 h of incubation with DC6, expressive disorganization of the cytoskeletal filaments was observed, more specifically of the actin filaments, showing an effect dependent on the concentration of DC6. In a three- dimensional model of in vitro metastasis, DC6 inhibited the formation of cellular spheroids in the neuroblastoma cell line SH-SY5Y. Thus it has been shown that DC6 has a double effect when it comes to tumor modulation, as it presents, in sub-toxic concentrations, the ability to induce cell death and reduce the migration of tumor cells via Nav1.7 modulation, as well as showing low toxicity in non-carcinogenic in vivo and in vitro models. Editor: Universidade Federal da Paraíba Tipo: Tese 2023-02-09T00:00:00Z https://repositorio.ufpb.br/jspui/handle/123456789/37667 Título: Bioprospecção de fungos filamentosos para produção de proteases utilizando leite desnatado como substrato Autor(es): Silva, Geisi Maria Henrique da Orientador: Amaral, Ian Porto Gurgel do Abstract: Filamentous fungi are highly relevant organisms in the production of bioactives. An example of bioactive compounds are enzymes, such as proteases, involved in important metabolic changes, which stand out in their use in industrial processes in several areas. However, the need to search for enzymes with more attractive characteristics for the industry and with low production cost is increasingly seen. In view of this, the present work aimed at the bioprospecting of filamentous fungi for the production of protease by submerged fermentation using skimmed milk with inducing substrate. Fungal isolates from zebrafish eggs were subjected to macroscopic (growth in Petri dish) and microscopic (cover slip culture) analysis, and the potential for proteolytic production in solid agar-milk medium, which proved to belong to the genera Penicillium sp. and Clonostahcys sp. and show positive protease activity. The isolates of Beauveria brongniartii and Paecilomyces sp., Were previously analyzed. For the production of protease, 0.5 ml of conidia suspension (1x109 conidia / ml) was inoculated in 50 ml of skimmed milk (5%), for 5 days at 180 rpm, RT. Proteolytic activity was determined using the azocazein and tricloacetic acid (TCA) method. The crude extracts from the fermentation processes showed activities of 1.3 U / mL for Clonostachys sp, 4.6 U / mL for Penicillium sp., 6.9 U / mL for Beauveria brongniartii and 30.3 U / mL for Paecilomyces sp. The crude extract that showed greater activity was subjected to partial purification using ethanol (0-80%). Where there was no significant increase in purity. The fraction purified by organic solvent with the highest total enzymatic activity (5.688 U), was subjected to purification by Ion Exchange Chromatography on a DEAE-Cellulose column, with a flow of 1mL / min and a mobile phase of 0.1M Tris-HCl pH 8 , 0. The purified enzyme showed 35% yield and a purification rate of 16.08 times. After purification, the protease from Paecilomyces sp. was characterized in terms of class, optimal temperature, thermostability and enzymatic kinetics. Serine protease inhibitors from which no inhibition was observed were used, demonstrating that the protease produced does not belong to this class. The enzymatic activity was analyzed at different temperatures (25, 30, 35, 40, 45, 50, 55 and 60 ° C), showing greater activity at 50 ° C, for the crude extract and 35 ° C, for the purified enzyme. As for thermal stability, the protease of the crude extract, after being incubated at temperatures 25; 35; 40; and 45 ° C, demonstrated greater stability in the first 2h, at 45 ° C. The kinetic parameters were found experimentally, using different concentrations of azocasein - 0.1 to 3.0% (w / v) -, of which the values of Km = 0.142% and Vmax = 6.8 U / mL were seen. In view of the results obtained, it is concluded that the fungi used were capable of producing protease by submerged fermentation and that the skimmed milk medium is a potential substrate for the production of proteolytic enzymes. Editor: Universidade Federal da Paraíba Tipo: Dissertação 2019-11-30T00:00:00Z