Leonardo Oliveira Trivilin

Toxicological evaluation of acute and sub-chronic ingestion of hydroalcoholic extract of Solanum cernuum Vell. in mice

ETHNOPHARMACOLOGICAL RELEVANCE Solanum cernuum Vellozo is a Brazilian shrub or small tree, restricted to Southwest states of the country. It has been widely used for the treatment of many ailments. The pharmacological activity of the extract on gastric ulcer has been the major therapeutic target proposed by the population investigated. MATERIALS AND METHODS In the acute toxicity test was used increasing doses of the extract (2, 4, 8, 12, 16, 20 and 25 g of extract per kilogram of body weight). The animal behavior was observed from 5h after a single administration of the extract and subsequently monitored daily until the fourteenth day, beyond the calculation of the estimated LD50 of the extract. In the test sub-chronic toxicity was used two doses of the extract (0.1 and 1.4 g/kg) and the parameters analyzed over 31 days were: body weight, food intake, behavior, respiratory rate, movement and mortality of animals. After anesthesia, blood samples were collected for hematological and biochemical analysis. The animals were euthanized followed by macroscopic analysis of the stomach and intestine. Liver, lungs and kidneys were removed, weighed and analyzed histopathologically. RESULTS In the acute toxicity test was observed a dose-dependent mortality and the value of estimated LD50 was 14.50 g/kg. In the hematological and biochemical analyses there were significant increase in the activities of AST and ALT indicating liver toxicity, but the extract was not able to alter food intake, body weight and organ weights after 31 days of treatment and it did not produce significant histopathological changes. CONCLUSION Therefore we can consider the hydroalcoholic extract of Solanum cernuum Vell as practically non-toxic in acute administration and safe in the sub-chronic administration, as hepatotoxicity was observed only with the highest dose used, not with the dose routinely used by the native population.

HIF-1alpha Expression Profile in Intratumoral and Peritumoral Inflammatory Cells as a Prognostic Marker for Squamous Cell Carcinoma of the Oral Cavity

The HIF-1 transcriptional complex is responsible for controlling transcription of over 100 genes involved in cell hypoxia response. HIF-1alpha subunit is stabilized in hypoxia conditions, creating the HIF-1 nuclear transcription factor. In inflammatory cells, high HIF-1alpha expression induces lymphocytic immunosuppression, decreasing tumoral antigen recognition, which promotes tumor growth. The present work investigated the relationship between HIF-1alpha expression in lymphocytes populating the intratumoral and peritumoral region of 56 patients with oral cancer. Our data indicates a prognostic value for this expression. High HIF-1alpha expression in peritumoral inflammatory cells is significantly related to worse patient outcome, whereas high expression in the intratumoral lymphoid cells correlates with a better prognosis. A risk profile indicating the chance of disease relapse and death was designed based on HIF-1alpha expression in tumoral inflammatory cells, defining low, intermediate and high risks. This risk profile was able to determine that high HIF-1alpha expression in peritumoral cells correlates with worse prognosis, independently of intratumoral expression. Low HIF-1alpha in tumor margins and high expression in the tumor was considered a low risk profile, showing no cases of disease relapse and disease related death. Intermediate risk was associated with low expression in tumor and tumor margins. Our results suggest that HIF-1alpha expression in tumor and peritumoral inflammatory cells may play an important role as prognostic tumor marker.

JMJD1A, H3K9me1, H3K9me2 and ADM expression as prognostic markers in oral and oropharyngeal squamous cell carcinoma

Aims Jumonji Domain-Containing 1A (JMJD1A) protein promotes demethylation of histones, especially at lysin-9 of di-methylated histone H3 (H3K9me2) or mono-methylated (H3K9me1). Increased levels of H3 histone methylation at lysin-9 (H3K9) is related to tumor suppressor gene silencing. JMJD1A gene target Adrenomeduline (ADM) has shown to promote cell growth and tumorigenesis. JMJD1A and ADM expression, as well as H3K9 methylation level have been related with development risk and prognosis of several tumor types. Methods and results We aimed to evaluate JMJD1A, ADM, H3K9me1 and H3K9me2expression in paraffin-embedded tissue microarrays from 84 oral and oropharyngeal squamous cell carcinoma samples through immunohistochemistry analysis. Our results showed that nuclear JMJD1A expression was related to lymph node metastasis risk. In addition, JMJD1A cytoplasmic expression was an independent risk marker for advanced tumor stages. H3K9me1 cytoplasmic expression was associated with reduced disease-specific death risk. Furthermore, high H3K9me2 nuclear expression was associated with worse specific-disease and disease-free survival. Finally, high ADM cytoplasmic expression was an independent marker of lymph node metastasis risk. Conclusion JMJD1A, H3K9me1/2 and ADM expression may be predictor markers of progression and prognosis in oral and oropharynx cancer patients, as well as putative therapeutic targets.

PAI-1, CAIX, and VEGFA expressions as prognosis markers in oral squamous cell carcinoma

BACKGROUND In oral squamous cell carcinoma (OSCC), the HIF-1 complex promotes the expression of genes involved in specific mechanisms of cell survival under hypoxic conditions, such as plasminogen activator inhibitor-1 (PAI-1), carbonic anhydrase 9 (CAIX), and vascular endothelial growth factor A (VEGFA). The study aimed to investigate the presence and prognostic value of PAI-1, CAIX, and VEGFA in OSCC. MATERIALS AND METHODS Immunohistochemistry was used to analyze the expressions of these proteins in 52 tumoral tissue samples of patients with OSCC, surgically treated and followed by a minimum of 24 months after surgery. The correlations between protein expressions and clinicopathological parameters and prognosis were analyzed. RESULTS Positive PAI-1 membrane expression was significantly associated with local disease relapse (P = .027). Multivariate analysis revealed that the positive PAI-1 membrane expression is an independent marker for local disease relapse, with approximately 14-fold increased risk when compared to negative expression (OR = 14.49; CI = 1.40-150.01, P = .025). Strong PAI-1 cytoplasmic expression was significantly associated with the less differentiation grade (P = .027). Strong CAIX membrane expression was significantly associated with local disease-free survival (P = .038). Positive CAIX cytoplasmic expression was significantly associated with lymph node affected (P = .025) and with disease-specific survival (P = .022). Multivariate analysis revealed that the positive CAIX cytoplasmic expression is an independent risk factor for disease-related death, increasing their risk approximately 3-fold when compared to negative expression (HR = 2.84; CI = 1.02-7.87, P = .045). Positive VEGFA cytoplasmic expression was significantly associated with less differentiation grade (P = .035). CONCLUSION Our results suggest a potential role for these expressions profiles as tumor prognostic markers in OSCC patients.

Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica

The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p > 0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.