Myocardium tissue changes caused by electrical transthoracic discharges in rats

<p>Abstract</p> <p>Background</p> <p>Cardiomyocytes cytoarchitecture changes caused by transthoracic countershocks have been focused recently. We aimed to evaluate the effects of electrical discharge application in the mitochondria structure in atrial myocardium of rats.</p> <p>Methods</p> <p>An electrical cardioverter was adapted to small rodent animals for our research. Electrical discharges were applied to the precordial region of 30 albino rats: (1) control group - animals that remained on resting period and were afterwards sacrificed; (2) electrical discharge group - animals that remained on resting period, followed by ten electrical discharges of 300 mV and sacrificed, and; (3) electrical post-discharge group - animals that remained on a resting period and received ten electrical discharges like the electrical discharge group, but were sacrificed seven days subsequently. We examined liver, adrenal and left atrium tissue fragments of the three groups.</p> <p>Results</p> <p>It was observed in control and post-discharge groups a normal cellular structure aspect with preserved architecture of cardiomyocytes and continuous sarcoplasmic membrane integrity. On the other hand, cardiac muscle fibers with mitochondrial edema and lysis occurred in the discharge group. Glycogen and adrenal lipids were not depleted in all groups.</p> <p>Conclusion</p> <p>These data suggest that transthoracic electrical discharges induce mitochondrial injuries in atrial cardiac cells of rats.</p

Memantine Prevents Cardiomyocytes Nuclear Size Reduction in the Left Ventricle of Rats Exposed to Cold Stress

OBJECTIVES: Memantine is an N-methyl-d-aspartate (NMDA) glutamate receptor antagonist used to treat Alzheimer's disease. Previous studies have suggested that receptor blockers act as neuroprotective agents; however, no study has specifically investigated the impact that these drugs have on the heart. We sought to evaluate the effects of memantine on nuclear size reduction in cardiac cells exposed to cold stress. METHOD: We used male EPM-Wistar rats (n=40) divided into 4 groups: 1) Matched control (CON); 2) Memantine-treated rats (MEM); 3) Rats undergoing induced hypothermia (IH) and 4) Rats undergoing induced hypothermia that were also treated with memantine (IHM). Animals in the MEM and IHM groups were treated by oral gavage administration of 20 mg/kg/day memantine over an eight-day period. Animals in the IH and IHM groups were submitted to 4 hours of hypothermia in a controlled environment with a temperature of - 8ºC on the last day of the study. RESULTS: The MEM group had the largest cardiomyocyte nuclear size (151 ± 3.5 &#956;m³ vs. CON: 142 ± 2.3 &#956;m³; p<0.05), while the IH group had the smallest mean value of nuclear size. The nuclear size of the IHM group was preserved (125 ± 2.9 &#956;m³) compared to the IH group (108 ± 1.7 &#956;m³; p<0.05). CONCLUSION: Memantine prevented the nuclear size reduction of cardiomyocytes in rats exposed to cold stress

RKIP Inhibition in Cervical Cancer Is Associated with Higher Tumor Aggressive Behavior and Resistance to Cisplatin Therapy

Cervical cancer is one of the most common cancers in women worldwide, being high-risk group the HPV infected, the leading etiological factor. The raf kinase inhibitory protein (RKIP) has been associated with tumor progression and metastasis in several human neoplasms, however its role on cervical cancer is unclear. In the present study, 259 uterine cervix tissues, including cervicitis, cervical intraepithelial lesions and carcinomas, were analyzed for RKIP expression by immunohistochemistry. We found that RKIP expression was significantly decreased during malignant progression, being highly expressed in non-neoplastic tissues (54% of the samples; 73/135), and expressed at low levels in the cervix invasive carcinomas (∼15% (19/124). Following in vitro downregulation of RKIP, we observed a viability and proliferative advantage of RKIP-inhibited cells over time, which was associated with an altered cell cycle distribution and higher colony number in a colony formation assay. An in vitro wound healing assay showed that RKIP abrogation is associated with increased migratory capability. RKIP downregulation was also associated with an increased vascularization of the tumors in vivo using a CAM assay. Furthermore, RKIP inhibition induced cervical cancer cells apoptotic resistance to cisplatin treatment. In conclusion, we described that RKIP protein is significantly depleted during the malignant progression of cervical tumors. Despite the lack of association with patient clinical outcome, we demonstrate, in vitro and in vivo, that loss of RKIP expression can be one of the factors that are behind the aggressiveness, malignant progression and chemotherapy resistance of cervical cancer.</p

O impacto do diagnóstico citológico de atipiasi indeterminadas no sistema público de saúde

As alterações citológicas de significado indeterminado representam uma importante limitação diagnóstica nos programas de escrutíneo de lesões cérvico-vaginais. A introdução de métodos biomoleculares, como o sistema de captura híbrida para detecção de HPV de alto risco contribui para a otimização da conduta clínica dessas pacientes, indicando colposcopia com precisão. Objetivo: avaliar o significado de lesões de significado indeterminado com relação à infecção pelo HPV, com o uso do teste de DNA para HPV com o método da captura de híbridos II. Métodos: foram estudadas amostras de 236 casos consecutivos examinados no laboratório da DIGENE-BRASIL, de pacientes com diagnóstico citológico prévio de ASCUS. As amostras foram submetidas ao teste de captura híbrida para identificação de DNA-HPV de alto e baixo riscos. Resultados: dos 236 casos analisados, 183 (77,5%) foram negativos para o teste de captura híbrida, seis (2,6%) foram positivos para HPV de baixo risco e 47 (19,9%) foram positivos para HPV de alto risco. Conclusão : as amostras positivas para HPV de baixo risco representam uma pequena e não- onsiderável minoria de casos, provavelmente, transientes. Cerca de 20% dos casos foram positivos para HPV de alto risco e deverão ser encaminhados à colposcopia e biopsia, se necessário. Esses casos representam um grande potencial de progressão para lesões cervicais.In order to optimize the morphological analysis of the cases with uncertain diagnosis, we critically analyzed the cases with Atypia of Squamous Cells of Undertemined Significance (ASCUS) in cytological samples of uterine cervix collected in conventional smears (CS) and liquidbased preparations (LBC) an to correlate the findings with Hybrid Capture II (HC2) assay and biopsy. Objective: to evaluate the meanig of undetermined cytological atypia in relation to HPV infection detected by hybrid capture II test. Methods: 97 cases taken from women examined at Perola Byignton Hospital, São Paulo, Brazil, during the year of 2002. The conventional smears were taken previously than LBC. The residual sample was placed in liquid-medium and LBC preparation with DNA-Citoliq system was performed. If at least one of the paired samples were classified as ASCUS, the pair was submitted to a guided revision in order to evaluate the type of alteration taken in account to categorized ASCUS. Results: from 97 cases studied, 14 were categorized as ASCUS by the two methods simultaneously. The others had different classification under or hyper estimated. Six cases diagnosed as squamous intraepithelial lesion (SIL) by CS were ASCUS by LBC; in contrary, 19 ASCUS by CS were SIL by LBC. Eleven ASCUS by CS were diagnosed as negative by LBC, but CS categorized 47 LBC ASCUS as negative. From the morphological parameters nuclear enlargement and coarse chromatin were regarded as ASCUS. From 68 ASCUS by LBC, 36 were HC2 positive for high risk HPV (hr-HPV) : ten of them with biopsy proven lesion. From 42 CS ASCUS, 23 were hr-HPV positive, but only 7 with histological lesion. Conclusion: our results reinforced the hypothesis that ASCUS is poorly reproducible by morphological examination by CS or LBC preparations. To add HC2 as adjunct method to ASCUS cytology can improve the routine diagnosed of the uncertain atypies

The association of p16INK4A and fragile histidine triad gene expression and cervical lesions

Objective. This cross-sectional study was intended to assess the association between immunohistochemical analysis of p16INK4A and fragile histidine triad (FHIT) and the presence of precancerous cervical lesions. Materials and Methods. Women seen at Pe´ rola Byington Hospital, São Paulo, Brazil, with histologically confirmed cervicitis (n = 31), cervical intraepithelial neoplasia (CIN) 1 (n = 30), CIN 2,3 (n = 30), and cervical cancer (n = 7) had also cervical material collected for liquid-based cytology, human papillomavirus Hybrid Capture 2 (HC2) test, and p16 and FHIT immunohistochemical reactions. Results. p16 and FHIT reactions were scored as the following: G1%, 1% to 5%, 95% to 25%, and 925%. Receiver operating curve analysis was used to select p16 and FHIT score cutoffs for further categorical analyses. All but one of the 37 CIN 2,3/cancer cases had a p16 score of greater than 1% to 5%. Among the 61 cervicitis/CIN 1 cases, 46 (75%) had a p16 score lower than 1% to 5%. In contrast, no association of FHIT expression and severity of cervical lesions could be demonstrated in this data set. Receiver operating curve analyses suggested the score of 1% to 5% for p16 as the cutoff that best discriminates CIN 2,3/cancer from cervicitis/CIN 1. No cutoff for FHIT scores could be suggested with data set. Conclusions. p16, but not FHIT expression, has the potential to be used as complementary diagnostic tool to investigate human papillomavirusYinduced cervical lesions, if these results are confirmed in larger studies.(undefined

The step of incorporation of Bacillus coagulans GBI-30 6086 into “requeijão cremoso” processed cheese does not affect metabolic homeostasis of rats

Dairy product consumption is a common habit in Brazil. These products present a good matrix for probiotic incorporation. Thus, in this study the feasibility of producing a probiotic "requeijao cremoso" incorporated with Bacillus coagulans GBI-30 6086 in three different steps and its metabolic effect in an animal model for 2 weeks has been evaluated. Wistar adult health rats were randomized into one to five groups (n = 8 for each group): Control (C); "requeijao cremoso" without probiotic (RC); probiotic inoculated in the milk before pasteurization at 65 degrees C/30 min (RPP); "requeijao cremoso" inoculated before the fusion step and consequently exposed to 90 degrees C/5 min (RPF); and "requeijao cremoso" inoculated after fusion step, i.e., once the product temperature reached 50 degrees C (RPAF). At the end of treatment, analysis of molecular markers of proteins of stress and antioxidant system, HSP 25, 60, 70 and 90, SOD and catalase were performed in the animals' muscles by Western Blot technique. The HSP25, HSP90 and catalase levels of C, RPP, RPF, and RPAF were similar, indicating that the homeostasis remained unchanged. The incorporation of B. coagulans GBI-30 6086 in the "requeijao cremoso" was shown to be stable and the microorganism remained viable in all steps tested. The incorporation of the probiotic strain in the fusion stage facilitated the technological process, since it allowed a better homogenization of the product and did not affect the maintenance of the metabolic homeostasis of rats10CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação302763/2014-7; 305804/2017-013/21544-9; 18/24540-8; 2019/21188-

Screening for cervical Cancer in high-risk populations: DNA pap test or hybrid capture II test alone?

This study was designed to evaluate whether Hybrid Capture II (HC2) test alone refer women to colposcopy as appropriately as DNA Papanicolaou (Pap) test, in the context of a high-risk group of women using the recently validated DNACitoliq LBC system. Women with suspected cervical disease were included in this crosssectional study at a tertiary center in São Paulo, Brazil, for further workup. All women had cervical material collected for LBC and HC2 for high-risk human papillomavirus (hrHPV)-DNA test. Irrespective of cytology and HC2 results, colposcopy, and cervical biopsy when applicable, was systematically performed. All tests were performed blindly. Sensitivity, specificity, positive and negative predictive values, and overall accuracy of both methods were computed in relation to histology. A total of 1,080 women were included: 36.4% (393/1080) had ACUS+, 10.2% (110/1080) were high-grade squamous intraepithelial lesions (HSIL) or cancer. Mean age was 33.5 years. All women underwent colposcopy, and cervical biopsies were performed in 38.4% (415/1080): 33% (137/415) of the biopsies were negative, 14.4% (155/415) were low-grade squamous intraepithelial lesions (LSIL), 10.7% (116/415) were HSIL, and 0.6% (7/415) were cancer. HC2 sensitivity to diagnose biopsy-proven HSIL was 100%. Because all HSIL cases had a positive HC2 test, sensitivity could not be improved by adding LBC. Specificity and positive and negative predictive values of DNA Pap were not significantly different from HC2 test alone when considering LSIL+ histology as ‘‘gold standard’’ and HSIL+ histology. As a screening strategy for women with high-risk for cervical cancer, DNA Pap test does not seem to add substantially to HC2 alone in terms of appropriately referring to colposcopy

RKIP Inhibition in cervical cancer Is associated with higher tumor aggressive behavior and resistance to cisplatin therapy

Cervical cancer is one of the most common cancers in women worldwide, being high-risk group the HPV infected, the leading etiological factor. The raf kinase inhibitory protein (RKIP) has been associated with tumor progression and metastasis in several human neoplasms, however its role on cervical cancer is unclear. In the present study, 259 uterine cervix tissues, including cervicitis, cervical intraepithelial lesions and carcinomas, were analyzed for RKIP expression by immunohistochemistry. We found that RKIP expression was significantly decreased during malignant progression, being highly expressed in non-neoplastic tissues (54% of the samples; 73/135), and expressed at low levels in the cervix invasive carcinomas (,15% (19/124). Following in vitro downregulation of RKIP, we observed a viability and proliferative advantage of RKIP-inhibited cells over time, which was associated with an altered cell cycle distribution and higher colony number in a colony formation assay. An in vitro wound healing assay showed that RKIP abrogation is associated with increased migratory capability. RKIP downregulation was also associated with an increased vascularization of the tumors in vivo using a CAM assay. Furthermore, RKIP inhibition induced cervical cancer cells apoptotic resistance to cisplatin treatment. In conclusion, we described that RKIP protein is significantly depleted during the malignant progression of cervical tumors. Despite the lack of association with patient clinical outcome, we demonstrate, in vitro and in vivo, that loss of RKIP expression can be one of the factors that are behind the aggressiveness, malignant progression and chemotherapy resistance of cervical cancer.This work was partially supported by the Portuguese Fundacao para a Ciencia e Tecnologia (grant PTDC/SAU-TOX/114549/2009). Olga Martinho and Sara Granja were recipients of PhD fellowships (SFRH/BD/36463/2007 and SFRH/BD/51062/2010, respectively), and Filipe Pinto and Vera Miranda-Goncalves were recipients of research fellowships (UMINHO/BI/016/2011 and SFRH/BI/33503/2008, respectively), both from FCT, Portugal. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding received for this study