Oral resveratrol supplementation improves Metabolic Syndrome features in obese patients submitted to a lifestyle-changing program.

AIMS: The aim of the present study was to evaluate the oral resveratrol effects associated with diet and physical training changes on anthropometric and biochemical parameters. MAIN METHODS: 25 individuals aged from 30 to 60 years old; with Body Mass Index (BMI) ≥ 30 kg/m2 were included in the study. Following the primary evaluation (anthropometric and clinical), the patients were randomly divided into 2 groups: (1) Placebo: Physical activity program + Diet + Placebo; (2) Resveratrol: Physical activity program + Diet + Resveratrol (RVS) (250 mg/day) for three months. Anthropometric and biochemical parameters were evaluated at baseline and after the treatment period. KEY FINDINGS: The main findings showed that the resveratrol supplementation improved total cholesterol (TC), High-density Lipoprotein cholesterol (HDL-c), Very-low density Lipoprotein cholesterol (VLDL-c), urea, creatinine and albumin serum levels. SIGNIFICANCE: These findings indicate that this polyphenol may be an option to potentiate the beneficial effects induced by dietary and physical activity programs in the Metabolic Syndrome (MetS) treatment.

Effect of insulin-transferrin-selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation on in vitro bovine embryo development.

The aim of this study was to evaluate the effect of adding a combination of insulin, transferrin and selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation (IVM) and in vitro culture (IVC) on in vitro embryo production. To verify the effect of the supplements, cleavage and blastocyst rates, embryo size and total cell number were performed. Embryonic development data, embryo size categorization and kinetics of maturation were analyzed by chi-squared test, while the total cell number was analyzed by a Kruskal-Wallis test (P < 0.05). When ITS was present during IVM, IVC or the entire culture, all treatments had a cleavage and blastocyst rates and embryo quality, similar to those of the control group (P < 0.05). Supplementation of IVM medium with ITS and AA for 12 h or 24 h showed that the last 12 h increased embryo production (51.6%; n = 220) on D7 compared with the control (39.5%; n = 213). However, no improvement was observed in blastocyst rate when less competent oocytes, obtained from 1-3 mm follicles, were exposed to ITS + AA for the last 12 h of IVM, with a blastocyst rate of 14.9% (n = 47) compared with 61.0% (n = 141) in the control group. The results suggest that the addition of ITS alone did not affect embryo production; however, when combined with AA in the last 12 h of maturation, there was improvement in the quantity and quality of embryos produced. Furthermore, the use of ITS and AA during IVM did not improve the competence of oocytes obtained from small follicles.

Molecular markers for oocyte competence in bovine cumulus cells.

This study aimed to quantify the expression of candidate genes in cumulus cells (CCs) from cumulus-oocyte complexes (COCs) with high and low potential for in vitro development up to the blastocyst stage. First, the effects of individual culture and biopsy on embryo development were evaluated. Individuals cultured using the well of the well system were compared with individuals cultured in 20 µL droplets (microdroplets) and those cultured in groups (control). Blastocyst rates were lower for the individual culture systems (P < 0.05; well of the well = 17.9%, n = 95; microdrop = 26.3%, n = 95) than for the control group (45.0%, n = 209). Second, the effects of biopsy on embryo production were compared between the control and microdroplet cultures, and no effects (P > 0.05) were observed for either group. Finally, the expression profiles of glypican 4 (GPC4), IGF4-binding protein, follicle-stimulating hormonereceptor, growth hormone receptor, epidermal growth factor receptor, fibroblast growth factor 11, solute carrier family 2 member 1, solute carrier family 2 member 3,sprouty homolog 1, versican, and keratin protein 8 in CCs obtained by biopsy were quantified by real-time polymerase chain reaction. Cumulus cells were categorized on the basis of the fates of the COCs: expanded blastocyst, cleaved and arrested, and uncleaved. The GPC4 gene was overexpressed (P = 0.007) in CCs from oocytes that formed embryos compared with those that produced cleaved and arrested embryos. We concluded that individual culture reduced blastocyst production; however, biopsy did not affect embryo development. The profile of GPC4 expression can be used as a marker to distinguish COCs with potential for embryo development from those with limited developmental potential.

The effect of pre-maturation culture using phosphodiesterase type 3 inhibitor and insulin, transferrin and selenium on nuclear and cytoplasmic maturation of bovine oocytes.

This study aims to evaluate if a pre-maturation culture (PMC) using cilostamide as a meiotic inhibitor in combination with insulin, transferrin and selenium (ITS) for 8 or 24 h increases in vitro embryo production. To evaluate the effects of PMC on embryo development, cleavage rate, blastocyst rate, embryo size and total cell number were determined. When cilostamide (20 µM) was used in PMC for 8 or 24 h, 98% of oocytes were maintained in germinal vesicles. Although the majority of oocytes resumed meiosis after meiotic arrest, the cleavage and blastocyst rates were lower than the control (P 0.05) to the control. The deleterious effect of 20 µM cilostamide treatment for 24 h on a PMC was confirmed by lower cumulus cell viability, determined by trypan blue staining, in that group compared with the other groups. A lower concentration (10 µM) and shorter exposure time (8 h) minimized that effect but did not improve embryo production. More studies should be performed to determine the best concentration and the arresting period to increase oocyte competence and embryo development.

Evaluation of the simulated physiological oocyte maturation system for improving bovine in vitro embryo production.

The aim of this study was to test the simulated physiological oocyte maturation (SPOM)- adapted system during bovine oocyte maturation to improve embryo development. Oocytes were obtained from follicles of 3 to 8 mm in diameter that were aspirated from ovaries obtained from a slaughterhouse. To verify the effect of the maturation system on in vitro embryo production, the cleavage, blastocyst rates on Days 7 and 8, embryo size, and total cell number were evaluated. The resulting data on embryo development were analyzed by the chi-square test, whereas data on embryo size and total cell number were analyzed by the Kruskal-Wallis test. First, the SPOM system principle was tested in our IVM system, in which 0.01 IU/mL of purified FSH and 10% of fetal calf serum were used during maturation. However, the cleavage and blastocyst rates on Days 7 and 8 were drastically reduced compared with those of the control group (P < 0.05). Increasing the dose of purified FSH to 0.1 IU/mL in the SPOM-adapted system did not affect (P > 0.05) embryo production, which remained lower than that of the control group. When less competent oocytes obtained from 1 to 3 mm follicles were used, the SPOM-adapted system was also unable to improve embryo production. To make the adapted system as similar as possible to the reported system, recombinant FSH was associated with BSA during maturation and embryo culture was performed under low oxygen tension conditions. Nevertheless, a reduction (P < 0.05) in the blastocyst rates was also observed, whereas the size and total cell number were similar to those of the control group (P > 0.05). It can be concluded that an SPOM-adapted system used under different culture conditions does not improve in vitro embryo development.

Fluoxetine reduces periodontal disease progression in a conditioned fear stress model in rats.

BACKGROUND AND OBJECTIVE: Recent evidence suggests that the use of fluoxetine could reduce periodontal disease severity. However, the effect of fluoxetine on periodontal disease has not been tested in the context of conditioned fear stress (CFS). We hypothesized that inhibition of chronic stress by fluoxetine might decrease the levels of bone loss in periodontal disease. The aim of the present study was to analyze the effect of fluoxetine on bone loss in chronic periodontitis. MATERIAL AND METHODS: Fourteen Wistar rats were submitted to ligature-induced periodontal disease and divided into four groups (A-D). Groups A (n = 3) and B (n = 4) were not stressed, while Groups C (n = 3) and D (n = 4) were submitted to a CFS paradigm for 38 d. Daily fluoxetine (20 mg/kg) was administered to Groups B and D from day 20 to day 39, at which point the rats were submitted to an open field test and killed on day 40. Mandibles were removed for histological and immunohistochemical analyses. RESULTS: Stress was associated with a higher level of bone loss in Group C compared with Group A. Additionally, no differences in bone loss were observed among Groups A, B and D. CONCLUSION: We showed that stress is associated with the progression of bone loss in a CFS model in rats and that fluoxetine treatment reduces the bone loss.

Diazepam reverses the alveolar bone loss and hippocampal interleukin-1beta and interleukin-6 enhanced by conditioned fear stress in ligature-induced periodontal disease in rats.

BACKGROUND AND OBJECTIVE: Stress and anxiety have been associated with chronic periodontitis, but few studies examining the effects of psychotropic drugs on periodontal health have been performed. Therefore, we aimed to investigate the effects of diazepam on the progression of periodontitis in chronically stressed rats. MATERIAL AND METHODS: Fourteen Wistar rats were submitted to ligature-induced periodontal disease and were divided into four groups . Two groups were not stressed, whereas two groups were submitted to a conditioned fear stress paradigm for 38 d. Daily diazepam treatment (2 mg/kg, orally) was administered to one unstressed group and to one group submitted to a conditioned fear stress paradigm from day 2 to the day 39, at which point the rats were submitted to an open field test and were killed on day 40. Brains and mandibles were removed for histological and immunohistochemical analyses. RESULTS: Animals exposed to conditioned fear stress presented an increase in freezing behavior, a decrease in locomotor activity, enhanced alveolar bone loss and higher levels of hippocampal interleukin-1beta (IL-1ß) and interleukin-6 (IL-6), compared with the control group. Diazepam, at the dose used in the current study, had no effect on freezing behavior but reversed the decrease in locomotor activity provoked by stress. Additionally, the treatment reduced the levels of hippocampal IL-1ß and IL-6 and alveolar bone loss in Wistar rats. Neither conditioned fear stress nor diazepam treatment had an effect on periodontal IL-1ß or IL-6 levels in animals. CONCLUSION: Our results suggest that diazepam treatment reduces bone loss in rats submitted to conditioned fear stress. In addition, diazepam treatment led to decreased IL-1ß and IL-6 levels in the hippocampus.

Detection and quantification of mast cell, vascular endothelial growth factor, and microvessel density in human inflammatory periapical cysts and granulomas.

AIM: To identify and quantify mast cell (MC), vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in human periapical cysts and granulomas. METHODOLOGY: Archived samples of cysts (n = 40) and granulomas (n = 28) were sectioned and stained with toluidine blue. MCs were identified and counted. Immunohistochemical reactions were employed to evaluate the tissue expression of VEGF and vessels. MVD was estimated by determining the areas of tissue labelled with CD31 antibody. The data were analysed using the Mann-Whitney test (P < 0.05). RESULTS: MCs were observed in the peripheral regions of both lesion types, whilst VEGF and MVD were distributed in the stroma. The presence of MCs was higher in cysts than in granulomas (P < 0.05). VEGF and MVD expression were similar in these lesions. CONCLUSIONS: The highest number of MCs was observed in cysts. Moreover, the identification of VEGF and MVD was consistent with the immune mechanisms involved in the lesions.

Lip squamous cell carcinoma in a Brazilian population: epidemiological study and clinicopathological associations.

OBJECTIVES: It was evaluated epidemiological aspects of primary lip squamous cell carcinoma (LSCC) and its associations with clinicopathological factors. STUDY DESIGN: This retrospective, cross-sectional study analysed a socio-demographic, clinical, and morphological data of HNSCC in a Brazilian population (n=30). Data analysis included descriptive statistics and bivariate analyses using the chi-square and Fisher 's exact tests to compare the variables. RESULTS: The LSCC represented 10.8% of all oral cavity squamous cell carcinoma. Lip malignant disease was more prevalent in elderly men, with male-to-female ratio of 5:1. Lower lip was more affected. It was observed high rates of chronic solar exposure, and tobacco and alcohol drinking habits. Clinically, early TNM staging, small tumour lesions, and non-metastatic disease were predominant findings. It was identified a high frequency of well differentiated tumor samples. Worse Karnofsky performance status was associated with cervical metastasis. CONCLUSIONS: Our findings showed that LSCC patients exhibited similar epidemiological and clinical profiles as noted in other studies. Still, the occurrence of metastatic disease was associated with a worse physical performance status of the LSCC patients during diagnosis.

Idiopathic gingival fibromatosis: description of two cases.

Gingival fibromatosis is an enlargement localized or generalized of the gingival tissue characterized by an expansion and accumulation of the connective tissue, predominantly type I collagen, with occasional presence of increased number of cells, supposed fibroblastic proliferation. Gingival fibromatosis can be induced as a side effect of systemic drugs, such as phenytoin, cyclosporin, and nifedipine, or due to hereditary factors. However, in some cases, the gingival overgrowth is idiopathic. This paper reports two cases of idiopathic gingival fibromatosis and discusses the diagnosis, histopathological features, treatment and immunohistochemical evaluation of myofibroblasts of this condition. The tissues removed were fixed in formalin, and sections used for hematoxylin and eosin and Masson tricromic stain. To determine the presence of myofibroblasts, we performed immunohistochemistry against a-SMA protein. Histological examination revealed epithelial hyperplasia with long rete pegs and increase in the dense fibrous connective tissue. The Masson tricromic stain revealed wide bundles of collagen strongly stained. It was showed negative labeling to a-SMA. These results strongly suggest that myofibroblasts are not involved in gingival overgrowth in the cases of IGF reported. Future studies will be necessary to determine the pathogenesis of idiopathic gingival fibromatosis.

Analysis of 724 cases of primary head and neck squamous cell carcinoma (HNSCC) with a focus on young patients and p53 immunolocalization.

This study evaluated 724 primary head and neck squamous cell carcinoma (HNSCC) in young and old patients, with regard to clinical profile and immunohistochemical expression of p53 protein. Associations among age, epidemiological and clinicopathological parameters, and survival analysis were evaluated. HNSCC in young people occurred in 14.5% (median age 40.7years; male-to-female ratio 5.9:1). A statistical association was demonstrated between age and family history of cancer, and between age and anatomical site. Among older patients, a higher presence of disease was noted in posterior sites. Expression of p53 was found in 71.7% of the samples and a higher expression was noted in lesions of young patients. Survival analysis showed that the age parameter is not a reliable prognostic factor for HNSCC. Among young patients, cervical metastasis was associated with worse survival. The presence of a family history of cancer in young patients could indicate genetic susceptibility and molecular disturbances in the p53 pathway in HNSCC of young and older patients seem to be distinct.

Cytomegalovirus shedding in the oral cavity of allogeneic haematopoietic stem cell transplant patients.

OBJECTIVE: This study was designed to investigate the effect of allogeneic haematopoietic stem cell transplantation (HSCT) on cytomegalovirus (CMV) shedding in the saliva by nested polymerase chain reaction (nested PCR) and its impact on patient survival. PATIENTS AND METHODS: One hundred and twenty-four HSCT patients and 124 healthy volunteers were included in the study. Oral swabs were taken before, after 100 days and 1 year of HSCT at the buccal mucosa. Nested PCR was used to detect CMV in the saliva. Time of death after HSCT was displayed, by means of the Kaplan-Meier method, for the following parameters: age and gender of the patient, donor gender, primary disease, stem cell source, platelet number, chronic graft vs host disease (cGVHD) of salivary glands and oral mucosa, and oral CMV shedding. Cox proportional hazards model was used for multivariate survival analysis. RESULTS: While none of the individuals in the control group showed positive swabs for CMV, the frequency of positive CMV oral swabs in patients at day + 100 after HSCT (45.2%) was statistically higher than before (7.2%) and 1 year after HSCT (17.5%). The presence of CMV was not associated with cGVHD and did not have any impact on post-transplant survival. CONCLUSIONS: The present study shows that oral CMV shedding occurs after HSCT, especially at day +100 post-transplant. Identification of CMV in the saliva might be important for the early diagnosis of CMV infection in allo-HSTC.

Oral phaeohyphomycosis.

This is the first description of solitary phaeohyphomycosis affecting the mucosal surface. The lesion developed in the inferior lip of a 57-year-old woman. After surgical resection, histopathological examination evidenced characteristic brownish fungal structures within granulomatous-purulent inflammation. Amplification and sequencing of rDNA obtained from paraffin-embedded tissue identified Alternaria species, as the causative agent.

Association of interleukin-1beta polymorphism with recurrent aphthous stomatitis in Brazilian individuals.

BACKGROUND: Recurrent aphthous stomatitis (RAS) is characterized by recurrent episodes of oral ulceration in an otherwise healthy individual. Some reports in the literature indicate that RAS may have immunological, psychological, genetic and microbiological bases. The purpose of the present study was to investigate the possible association between interleukin-1beta (IL-1beta) +3954 (C/T) genetic polymorphism and RAS in a sample of Brazilian patients. SUBJECTS AND METHODS: Sixty-two consecutive subjects affected by minor and major forms of RAS and 62 healthy volunteers were genotyped at IL-1beta (+3954). The chi-squared test was used for statistical analysis. RESULTS: A significant increase in the high production of IL-1beta genotype CT was observed in the group with RAS (P = 0.01). After stratifying RAS patients according to the mean number of lesions per episode, a significant difference was only observed between patients with >or=3 lesions in each episode and control. CONCLUSION: There is an increased frequency of polymorphism associated with high IL-1beta production in RAS patients.

Herpes simplex virus type 1 shedding in the oral cavity of seropositive patients.

OBJECTIVE: Investigate the frequency of herpes simplex virus type 1 (HSV-1) reactivation in the oral cavity of seropositive patients with previous history of recurrent herpes labialis (recrudescent group) compared with those without any history of recrudescent lesions (asymptomatic HSV-1 infection). In addition, the relation between recrudescence and the presence of the virus in the saliva was assessed. MATERIALS AND METHODS: Fourteen individuals with previous history of herpes labialis (recrudescent group) and 11 HSV-1 seropositive asymptomatic volunteers were included in the study. Swabs were performed periodically in all subjects and the presence of HSV-1 DNA was identified by nested PCR. RESULTS: All the 25 subjects enrolled in the study, revealed at least one positive swab for HSV-1. The frequency of HSV-1 positivity in the group with recrudescent herpes labialis was not statistically different from the other group. Ten subjects of the recrudescent group presented with herpes labialis at least once during the study. CONCLUSIONS: HSV-1 shedding in the oral cavity occurs independently of herpes labialis recrudescence.

Effect of bone marrow transplantation on the immunolocalization of p53, hMSH2, and hMLH1 proteins on oral mucosa.

OBJECTIVE: Considering that hMSH2, hMLH1 and p53 are important in maintaining genomic stability of the oral mucosa epithelium, the purpose of the present study was to investigate the immunolocalization of these proteins in the epithelium of the oral mucosa of patients submitted to bone marrow transplantation (BMT) compared with controls. MATERIALS AND METHODS: Twenty-one samples of lip biopsies from BMT recipients were retrieved. Twenty samples of normal lower labial mucosa associated with mucocele in non-transplanted patients were included as control group. The streptavidin-biotin complex stain was used to detect the human DNA mismatch repair proteins hMSH2, hMLH1 and p53 protein. RESULTS: The main findings demonstrated that the mean number of suprabasal epithelial cells positive for MSH2 was statistically higher than the control group. The immunostaining of hMLH1 and p53 at the basal and suprabasal epithelial layers were statistically higher in the oral labial mucosa of the BMT patients compared with controls. CONCLUSION: The present study shows that oral epithelial cells of BMT patients show increased immunolocalization of the DNA repair related proteins.