Cinthia Gandolfi Boer

Production of extracellular protease by Aspergillus tamarii

The production of protease by Aspergillus tamarii was investigated in submerged fermentation using different substrates. Low basal proteolytic activity was detected on cultures with casein or gelatin as the only substrate (16–20 U/ml). The enrichment of protein medium with glucose increased the protease production up to 6 times (110 U/ml). The highest proteolytic levels were obtained in cultures supplemented with wheat bran (120 U/ml) and soybean meal (161 U/ml). One major proteolytic band with estimated molecular mass of 48 KDa was detected by SDS‐PAGE. The acetone‐precipitated enzyme was active and stable over a wide range of pH (6–9.5), and temperature (30–55 °C). The enzyme was stable for more than ten hours at temperatures up to 45 °C. The half‐lives at 50 and 55 °C were 90 min and 12 min, respectively. In presence of 5.0 mM CaCl2, the half‐life of the enzyme at 55 °C was increased to 130 min.

Evaluation of the Efficacy of Flaxseed Meal and Flaxseed Extract in Reducing Menopausal Symptoms

The objective of this study was to evaluate the efficacy of flaxseed meal and flaxseed extract in reducing climacteric symptoms of menopausal women. Ninety menopausal women were randomly distributed into three study groups: group I received 1 g per day of flaxseed extract containing at least 100 mg of secoisolariciresinol diglucoside (SDG), group II received 90 g per day of flaxseed meal containing at least 270 mg of SDG, and group III received 1 g per day of collagen (placebo group). Subjects were assessed for menopausal symptoms by the Kupperman index at the beginning and at the end of the 6 months of treatment. Subjects were also assessed for endometrial thickness and vaginal cytology. The Kupperman index values at the beginning and end of the treatments were analyzed using the paired t-test. Both the flaxseed extract (P=.007) and the flaxseed meal (P=.005) were effective in reducing the menopausal symptoms when compared with the placebo control (P=.082). Alternatively, the changes in Kupperman index were also computed and submitted to analysis of variance. In this case, no significant differences were found (P=.084) although the data indicate a decreasing tendency for the Kupperman index by both the flaxseed extract and the flaxseed meal groups. Neither the flaxseed extract nor the flaxseed meal exerted clinically important estrogenic effects on the vaginal epithelium or endometrium as revealed by the absence of changes in the blood levels of follicle stimulating hormone and estradiol, as well as in the endometrial thickness, and vaginal epithelial maturation value. No serious adverse events related to the treatments were reported. Although the results of the present study do not allow an unequivocal conclusion about the action of flaxseed on the menopausal symptoms, they suggest that it could be premature to conclude that no such action exists. Clearly the matter still deserves further experimental attention.

Human leukocyte antigen (HLA)-G and cervical cancer immunoediting: A candidate molecule for therapeutic intervention and prognostic biomarker?

While persistent infection with oncogenic types of human Papillomavirus (HPV) is required for cervical epithelial cell transformation and cervical carcinogenesis, HPV infection alone is not sufficient to induce tumorigenesis. Only a minor fraction of HPV infections produce high-grade lesions and cervical cancer, suggesting complex host-virus interactions. Based on its pronounced immunoinhibitory properties, human leukocyte antigen (HLA)-G has been proposed as a possible prognostic biomarker and therapeutic target relevant in a wide variety of cancers and viral infections, but to date remains underexplored in cervical cancer. Given the possible influence of HLA-G on the clinical course of HPV infection, cervical lesions and cancer progression, a better understanding of HLA-G involvement in cervical carcinogenesis might contribute to two aspects of fundamental importance: 1. Characterization of a novel diagnostic/prognostic biomarker to identify cervical cancer and to monitor disease stage, critical for patient screening; 2. Identification of HLA-G-driven immune mechanisms involved in lesion development and cancer progression, leading to the development of strategies for modulating HLA-G expression for treatment purposes. Thus, this systematic review explores the potential involvement of HLA-G protein expression and polymorphisms in cervical carcinogenesis.

Production, purification and characterization of tannase from Aspergillus tamarii

The production of tannases by Aspergillus tamarii was evaluated in submerged cultures using tannic acid and gallic acid as substrates. Two tannases, designated as TAH I and TAH II were produced in gallic acid submerged cultures. TAH I, responsible for 70% of the total tannase activity was purified to apparent electrophoretic homogeneity with 18.35% yield. The enzyme is a homodimeric protein with molecular mass of 180 kDa and 40.5% of its weight corresponds to carbohydrates. TAH I exhibited optimal activity at 30°C and pH 5.5 and was stable over a large pH range (3.0 to 9.0) and at temperatures up to 40°C. With methyl gallate as substrate, the enzyme presented a K M of 0.77 mM and a V max of 682.8 U/mg proteins. The enzyme was inhibited by metal ions but showed relative resistance to organic solvents and surfactants. Since the enzyme is active over a wide range of pH and temperature, it is potentially useful in food and pharmaceutical industries. Key words : Aspergillus tamarii , enzyme purification, submerged culture, tannase.

Chá verde brasileiro (Camellia sinensis var assamica): efeitos do tempo de infusão, acondicionamento da erva e forma de preparo sobre a eficiência de extração dos bioativos e sobre a estabilidade da bebida

Studies on the Brazilian green tea (Camellia sinensis var assamica) are still scarce when compared with the great number of investigations conducted on the green tea produced in other countries. The purposes of the present study were to evaluate the effects of infusion time, mode of packaging (tea in bulk or in tea bags) and preparation on the extraction of bioactive compounds from the Brazilian green tea and on the stability of the beverage. The evaluated parameters were: the amounts of soluble solutes and phenolic compounds that were extracted as well as the antioxidant properties of the beverage using the DPPH method (2,2-diphenyl-1-picrylhydrazyl radicals). The results revealed that the use of the herb in bulk with 5 minutes stirring was the most adequate condition for the extraction of bioactive compounds from the green tea. Increases in the volume of the preparation without changes in the herb/water ratio increased the efficiency of the bioactive compounds extraction due to the fact that cooling a large volume of water is delayed in comparison to cooling smaller volumes. It can be said that the obtained beverages were stable for 24 hour at room temperature and in the refrigerator since no decrease in the antioxidant properties and significant changes in the contents of the major bioactive coumpounds, namely epigallocatechin gallate, epicatechin, catechin, and caffeine were detected.

Design and optimization of self-microemulsifying drug delivery system (SMEDDS) of felodipine for chronotherapeutic application

The objective of this study was to determine the effect of medroxyprogesterone acetate (DMPA) on the development and maintenance of Candida albicans in the vagina of oophorectomized Wistar rats. The animals were divided into negative control groups (NCG), which received injections of sterile saline; positive control groups (PCG), which were given injections of estradiol valerate; and progesterone groups (PG), which were given injections of Depo-Provera®. After one week of hormonal induction, vaginal infection by C. albicans was induced in all the groups and detected by vaginal yeast culture and Papanicolaou smear. In addition, scanning and transmission electron microscopy images were obtained to confirm the vaginal infection by yeast in PG. A difference in progesterone levels in PG was observed between the basal level and after hormonal induction (P<0.0001). In this group, 100% of the rats acquired vaginal infection in the first week, but did not maintain it until the third week. The pharmaceutical brand of DMPA was effective for inducing the metestrus or diestrus phase of the estrous cycle in rats, similar to the use of pure progesterone. In contrast to estrogen treatment, progesterone alone could not support an experimental vaginal infection by C. albicans for any significant period of time.

Association between Human Papillomavirus and Non-cervical Genital Cancers in Brazil: A Systematic Review and Meta-Analysis

Background: Human papillomavirus (HPV) infection is associated with cervical cancer; however, it is controversial whether it is involved in non-cervical genital cancers. Objective: This study aimed to evaluate articles on the prevalence of HPV in penile cancer, vulvar cancer, colorectal cancer, prostate cancer and anal canal cancer in studies conducted in Brazil. Methods: The study was conducted in accordance with the Preferred Reporting of Systematic Reviews and Meta-Analysis Statement. Comprehensive searches for HPV and cancer for the years 2006 to 2016 were conducted in two databases (PubMed and Web of Knowledge) and Google Scholar system. We also tracked the references of all eligible articles to identify additional non-captured publications through online surveys. Results: Eighteen studies, with a combined sample size of 1,552 patients were analyzed. The overall prevalence of HPV was 43% (95% CI: 36–51%; p < 0.001). The pooled prevalence of HPV in penile cancer was 42% (95% CI: 32–55%; p < 0.001), in colorectal cancer it was 67% (95% CI: 64–70%; p < 0.001) and in vulvar cancer 43% (95% CI: 34–55%; p < 0.001). HPV 16 was the most prevalent in all sites evaluated, with prevalence estimated at 54% (95% CI: 44–66%; p < 0.001), followed by genotypes 33 (21%; 95% CI: 17–28; p < 0.001), 6 (15%; 95% CI: 8–26%; p < 0.001), 11 (13%; 95% CI: 5–32%; p < 0.001) and 18 (12%; 95% CI: 7–22%; p < 0.001), respectively. The pooled prevalence of single infection was 82% and infection by multiple genotypes of HPV was 22%. Conclusion: Our study demonstrated a high prevalence of HPV in non-cervical genital cancers in Brazil, with predominance of genotype 16, providing evidence for the need for preventive and control measures to avoid future harm to the population.