A novel antifungal peptide, PcAFP (6.48 kDa, pI 8.83), was obtained from the culture supernatant of the fungus Penicillium crustosum. The gene encoding the PcAFP peptide was isolated based on its homologue in Penicillium chrysogenum, PgAFP. PcAFP is a small, cystine-rich peptide, and the mature peptide consists of 58 amino acid residues. The immature P. crustosum antifungal protein (AFP) showed 95.65% identity to the antifungal protein of P. chrysogenum, while the mature peptide showed 98.28% identity with PgAFP. Molecular modeling of the tertiary structure of the mature peptide revealed details of the conserved structure of the AFPs, such as the β-barrel motif stabilized by three disulfide bonds and the l-core motif. Analysis of the extract by 16% tricine SDS-PAGE showed a 6.9 kDa peptide, which was close to the predicted molecular mass of the mature peptide of 6.48 kDa. Assays of antimicrobial activity, performed by broth microdilution using the crude extract obtained from the culture medium, showed activity against Candida albicans. These results demonstrate the conservation of the PcAPF gene and the high level of identity with the PgAFP antifungal protein of P. chrysogenum. Given these structural and biochemical characteristics, PcAFP could be a potential candidate for future investigations that may aid in the development of new antifungal compounds.
ResumoOBJETIVO: Avaliar o desenvolvimento folicular em ratas Wistar com obesidade induzida por dieta de cafeteria (DCAF) submetidas à administração de losartan (LOS), um antagonista do receptor AT 1 da Angiotensina II. MÉTODOS: Aos 21 dias de vida, as ratas foram separadas aleatoriamente em dois grupos: controle (CTL), que recebeu ração padrão, e cafeteria (CAF), que recebeu a DCAF, altamente palatável e calórica. Aos 70 dias de vida, início da idade reprodutiva, animais do grupo CAF foram subdivididos em dois grupos (n=15/grupo): CAF, que recebeu água, e CAF+LOS, que recebeu 30 mg/kg de peso corporal (PC) de LOS por gavagem durante 30 dias. O grupo CTL também recebeu água por gavagem. Aos 100 dias de vida foi realizada a eutanásia dos animais e o PC e das gorduras retroperitoneal, perigonadal e subcutânea foi avaliado. Os ovários direitos foram retirados para contagem do número dos diferentes tipos de folículos ovarianos. As concentrações plasmáticas dos hormônios folículo-estimulantes (FSH), luteinizante (LH), prolactina (PRL) e progesterona foram avaliadas. Os resultados foram expressos como média±erro padrão da média. Para análise estatística, foi utilizado one-way ANOVA, seguido pelo pós-teste de Newman-Keuls (p<0,05). RESULTADOS: O PC e das gorduras, assim como o número de folículos antrais, foi elevado no grupo CAF em relação ao CTL. Todavia, as concentrações de FSH e LH foram mais baixas entre os animais CAF. A administração de LOS reduziu o PC e das gorduras retroperitoneal e subcutânea, bem como o número de folículos antrais. O tratamento com LOS atenuou a redução das concentrações de FSH e de LH. As concentrações de progesterona e PRL foram semelhantes entre os grupos estudados. CONCLUSÃO: O uso de LOS pode favorecer o desenvolvimento folicular em fêmeas obesas e pode possibilitar sua utilização como fármaco coadjuvante no tratamento da infertilidade associada à obesidade. Abstract PURPOSE:To evaluate the follicular development of female Wistar rats with obesity induced by the cafeteria diet, submitted to the administration of losartan (LOS), an antagonist of the AT 1 receptor of Angiotensin II. METHODS: At weaning (21 days of age), female Wistar rats were randomly divided, into two groups: control (CTL) that received standard chow and cafeteria (CAF) that received a cafeteria diet, a highly palatable and highly caloric diet. At 70 days of age, at the beginning of the reproductive age, animals of the CAF group were subdivided into two groups (n=15/ group): CAF, that received water, and CAF+LOS, that received LOS for 30 days. The CTL group also received water by gavage. At 100 days of age, the animals were euthanized and body weight (BW) as well as the retroperitoneal, perigonadal and subcutaneous fat weights were analyzed. The right ovaries were isolated for counting the number of primary, secondary, antral and mature follicles. Plasma levels of FSH, LH, prolactin and progesterone hormones were analyzed. The results were expressed as mean±standard error of the mean. Data were analyzed statistically by...
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