Investigation of crude methanolic extract from poison secreted by the Rhaebo gutattus on status redox antioxidant in mice

The study evaluated the antioxidant properties of a crude methanolic extract (CME) from Rhaebo guttatus poison in mice over a period of 7 and 30 days. The mice were divided into groups and treated with different concentrations of the extract (0; 8 µg mL-1; 16 µg mL-1 and 32 µg mL-1 or vehicle; 100 µL/animal/day; via gavage). The liver samples were analyzed for status redox parameters as catalase (CAT), glutathione-S-transferase (GST), reduced glutathione (GSH) and thiobarbituric acid reactive substances (TBARS). The results showed that the CME caused changes in the levels of various antioxidants and oxidative stress markers. At 7 days, there was an increase in TBARS levels (8 µg mL-1 dose) and GST activity (16 µg mL-1 dose), and a reduction in GSH levels (32 µg mL-1 dose) compared to the control group. At 30 days, TBARS and GSH levels returned to control values in the same period, but GSH increased (32 µg mL-1 dose) compared at 7 days; GST activity remained high after 30 days for 32 µg mL-1 dose compared other groups and time of treatment (7 days). Overall, the study suggests that the extract modulates antioxidant properties per se that can affect various markers of status redox in the liver of mice, mainly 16 µg mL-1 dose demonstrated to act under antioxidant enzymes in different times (7 or 30 days).

O papel da atenção primária na rede de atenção à saúde frente ao manejo da hanseníase: um estudo transversal / he role of primary carein the health care network in front of leprosy management: a cross-sectional study

Objetivou-se analisar o perfil demográfico e clínico dos casos de hanseníase e associar os fatores relacionadas às incapacidades físicas com a Atenção Primária à Saúde(APS)na perspectiva de Redes de Atenção à Saúde em Sinop, Mato Grosso. Trata-se de um estudo transversal, que percorreu três etapas analíticas: conhecimento do perfil epidemiológico; classificação da capacidade da Atenção Primária em coordenar as Redes de Atenção à Saúde; correlação dessa classificação com o número de casos novos, desfecho e o Grau de Incapacidade Física e correlação do número de casos novos e a média de cobertura da Atenção Primária por meio do coeficiente de correlação de Pearson. Foram considerados os casos novos de hanseníase notificados no período de 2014 a 2018. Dos 1.649 casos novos, 63% foram mulheres, entre 15 e 59 anos (83%),com até nove anos de estudo (47%),e negros (61%). Cura respondeu por 86% dos registros. Predomínio de Grau de incapacidade física0no diagnóstico (58%) e ausência de avaliação no desfecho (17,1%). Todas as Atenção Primária à Saúde foram classificadas como 'condição boa' na coordenação das Redes de Atenção. O abandono apresentou 'correlação moderada' com a capacidade da Atenção Primária em coordenar as Redes de Atenção e a média de cobertura da Atenção Primária com o número de casos indicou correlação 'muito alta'. A Atenção Primária, na perspectiva da Rede de Atenção, se mostrou efetiva no controle e na condução da hanseníase no território municipal.

The objective was to analyze the clinical epidemiological profile of leprosy cases and associate this profile with primary health care from the perspective of health care networks. This is a cross-sectional study that covered three analytical stages: knowledge of the epidemiological profile; classification of the capacity of the APS to coordinate the RAS; correlation of this classification withthe numberof new cases, outcome and the Degree of Physical Disability (GIF),and correlation of the number of new cases and the average coverage of PHC, using the Pearson correlation coefficient. New cases of leprosy reported in the period from 2014 to 2018 were considered. Of the 1,649 new cases, 63% were women, between 15 and 59years (83%), with up to nine years of education (47%) and black (61% ). Healing accounted for 86% of the records. The predominance of GIF 0 in the diagnosis (58%) and absence ofevaluation in the outcome (16%). All UBS were classified as 'good condition' in the RAS coordination. Dropout showed a 'moderate correlation' with the PHC's ability to coordinate the RAS and the mean PHC coverage with the number of cases indicated a 'veryhigh' correlation. APS, from the perspective of RAS, proved to be effective in controlling and managing leprosy in the municipal territory.

Microwave-assisted green synthesis of fluorescent carbon quantum dots from Mexican Mint extract for Fe3+ detection and bio-imaging applications.

Biomass-derived carbon quantum dots have drawn special interest owing to their admirable photostability, biocompatibility, fluorescence, high solubility, sensitivity and environmentally friendly properties. In the present work, the Carbon Quantum Dots (CQDs) was synthesized from the Plectranthus amboinicus (Mexican Mint) leaves via the microwave-assisted reflux method. The strong absorption peaks observed from UV-vis spectra at 291 and 330 nm corresponds to the π-π* and n-π* transitions, respectively, reveal the formation of CQDs. The synthesized CQDs showed bright blue fluorescence under UV irradiation with a fluorescence quantum yield of 17% and a maximum emission of 436 nm in the blue region at an excitation wavelength of 340 nm. The HRTEM analysis elucidates that the synthesized CQDs were crystalline and spherical in shape with a particle size of 2.43 ± 0.02 nm. The FT-IR spectroscopy confirms the presence of the different functional groups such as -OH, -CH, CO and C-O. The chemical composition of CQD was revealed through XPS analysis. The synthesized CQDs were used as a fluorescent probe to detect different metal ions, where high selectivity was obtained for Fe3+ ions through quenching phenomenon. The emission intensity of CQD showed a good linear relationship with R2 = 0.9111 with the concentration of Fe3+ ions in the range of 0-15 µM. The fluorescence emission of CQD was turned OFF upon the binding of Fe3+ ions and turned - ON with the addition of ascorbic acid. With this fluorescent turn ON-OFF behaviour of CQD, the NOT and IMPLICATION logic gates were constructed and studied for different input conditions. The biocompatibility of CQD was tested via MTT assay using MCF7 breast cancer cell line, which revealed that CQD synthesized from the Mexican Mint leaves possess less cytotoxicity. Further, the prepared CQD was applied effectively as fluorescent probes in a cell imaging application.

Optimization of Zika DNA vaccine by delivery systems.

In our previous study, we designed and evaluated the efficacy of six DNA vaccine candidates based on the E protein of Zika virus (ZIKV). To optimize the DNA vaccine, we inoculated C57BL/6 and IFNAR1- mice with the vaccine candidate expressing tandem repeated ZIKV envelope domain III (ED III × 3) doses; 50 µg by intramuscular (IM), jet injection (JET), or electroporation (EP) routes. Results showed that vaccination by all routes induced humoral and cellular immunity. Among them, EP induced robust ZIKV E specific-total IgG and neutralizing antibodies, as well as T cell responses. Additionally, EP showed superior protective efficacy against the ZIKV Brazil strain compared to the IM and JET routes. Finally, in the dose optimization test of EP route, cellular immunity of 50 µg was induced a significant level than other dose groups. These results showed that the EP delivery system enhanced the potential immunogenicity and protective efficacy of DNA vaccines.

NEK2 Promotes Hepatoma Metastasis and Serves as Biomarker for High Recurrence Risk after Hepatic Resection.

INTRODUCTION AND AIM: Developing reliable biomarkers for hepatocellular carcinoma (HCC) patients who are at a high risk of recurrence after curative hepatic resection is very important for determining subsequent therapeutic strategies. We investigated the role of the cell cycle factor NIMA-related kinase 2 (NEK2) in HCC progression in hepatoma cells and post-surgery patients. MATERIAL AND METHODS: The effects of NEK2 on proliferation, invasion and migration of hepatoma HuH7 and SK-Hep1 cells were evaluated. In a post-surgery HCC cohort (N = 97), the Nek2 induction levels in the tumors were examined with real-time RT-PCR analysis, and the results were analyzed for their correlations with recurrence. RESULTS: NEK2 promoted G1 to S phase cell cycle progression by causing increases in cyclin D1 and AKT phosphorylation and decreases in the cyclin-dependent kinase inhibitor p27, indicating that NEK2 plays an important role during interphase in addition to its previously identified role in M phase. NEK2 also enhanced the proliferation, migration and invasion of hepatoma cells and regulated the expression of E-cadherin and MMP9. The Nek2 mRNA levels in the tumors were highly correlated with recurrence rates in the post-surgery HCC patients. Combined evaluation of the tumor AJCC stage and the Nek2 level can serve as a reliable method for predicting the relative risk of HCC recurrence in these patients. CONCLUSIONS: NEK2 plays a significant role in cell cycle progression in the inter- and M-phases. NEK2 enhances HCC metastasis and is correlated with recurrence and thus can potentially serve a promising high-risk biomarker for HCC.

Loss of c-Met signaling sensitizes hepatocytes to lipotoxicity and induces cholestatic liver damage by aggravating oxidative stress.

Recent studies confirmed a critical importance of c-Met signaling for liver regeneration by modulating redox balance. Here we used liver-specific conditional knockout mice (MetKO) and a nutritional model of hepatic steatosis to address the role of c-Met in cholesterol-mediated liver toxicity. Liver injury was assessed by histopathology and plasma enzymes levels. Global transcriptomic changes were examined by gene expression microarray, and key molecules involved in liver damage and lipid homeostasis were evaluated by Western blotting. Loss of c-Met signaling amplified the extent of liver injury in MetKO mice fed with high-cholesterol diet for 30days as evidenced by upregulation of liver enzymes and increased synthesis of total bile acids, aggravated inflammatory response and enhanced intrahepatic lipid deposition. Global transcriptomic changes confirmed the enrichment of networks involved in steatosis and cholestasis. In addition, signaling pathways related to glutathione and lipid metabolism, oxidative stress and mitochondria dysfunction were significantly affected by the loss of c-Met function. Mechanistically, exacerbation of oxidative stress in MetKO livers was corroborated by increased lipid and protein oxidation. Western blot analysis further revealed suppression of Erk, NF-kB and Nrf2 survival pathways and downstream target genes (e.g. cyclin D1, SOD1, gamma-GCS), as well as up-regulation of proapoptotic signaling (e.g. p53, caspase 3). Consistent with the observed steatotic and cholestatic phenotype, nuclear receptors RAR, RXR showed increased activation while expression levels of CAR, FXR and PPAR-alpha were decreased in MetKO. Collectively, our data provide evidence for the critical involvement of c-Met signaling in cholesterol and bile acids toxicity.

The effect of muscle fatigue on the last stride before stepping down a curb.

The stride before landing may be important during stepping down. The aim of this study was to analyze variability of the kinematics and muscle activity in the final stride before stepping down a curb, with and without ankle and knee muscle fatigue. Ten young participants walked at self-selected speed and stepped down a height difference (10-cm) in ongoing gait. Five trials were performed before and after a muscle fatigue protocol (one day: ankle muscle fatigue, another day: knee muscle fatigue). The analysis focused on the trailing leg during the last but one and the last step on the higher level. Kinematics and muscle activity were recorded. Fatigue increased variability of foot-step horizontal distance in the last step on the higher level of the trailing limb, as well as in the first steps on the lower level for both limbs. This appeared due to an increase in the range of motion of the knee joint after both fatigue protocols. Participants additionally showed an increased ankle and hip ROM and decreased knee ROM. Our results suggest a loss of control under fatigue reflected in a higher variability of trailing and leading limb-step horizontal distances, with compensatory changes to limit fatigue effects, such as a redistribution of movement over joints.

Simvastatin acts as an inhibitor of interferon gamma-induced cycloxygenase-2 expression in human THP-1 cells, but not in murine RAW264.7 cell

Cyclooxygenase-2 (COX-2) is a key inflammatory response molecule, and associated with many immune functions of monocytes/macrophages. Particularly, interferon gamma (IFNgamma)-induced COX-2 expression appears in inflammatory conditions such as viral infection and autoimmune diseases. Recently, statins have been reported to show variable effects on COX-2 expression, and on their cell and species type dependences. Based on the above description, we compared the effect of simvastatin on IFNgamma-induced COX-2 expression in human monocytes versus murine macrophages. In a result, we found that simvastatin suppresses IFNgamma-induced COX-2 expression in human THP-1 monocytes, but rather, potentiates IFNgamma-induced COX-2 expression in murine RAW264.7 macrophages. However, signal transducer and activator of transcriptio n 1/3 (STAT1/3), known as a transcription factor on COX-2 expression, is inactivated by simvastatin in both cells. Our findings showed that simvastatin is likely to suppress IFNgamma-induced COX-2 expression by inhibiting STAT1/3 activation in human THP-1 cells, but not in murine RAW264.7 cells. Thus, we concluded that IFNgamma-induced COX-2 expression is differently regulated by simvastatin depending on species specific mechanism.

Simvastatin acts as an inhibitor of interferon gamma-induced cycloxygenase-2 expression in human THP-1 cells, but not in murine RAW264.7 cell

Cyclooxygenase-2 (COX-2) is a key inflammatory response molecule, and associated with many immune functions of monocytes/macrophages. Particularly, interferon gamma (IFNgamma)-induced COX-2 expression appears in inflammatory conditions such as viral infection and autoimmune diseases. Recently, statins have been reported to show variable effects on COX-2 expression, and on their cell and species type dependences. Based on the above description, we compared the effect of simvastatin on IFNgamma-induced COX-2 expression in human monocytes versus murine macrophages. In a result, we found that simvastatin suppresses IFNgamma-induced COX-2 expression in human THP-1 monocytes, but rather, potentiates IFNgamma-induced COX-2 expression in murine RAW264.7 macrophages. However, signal transducer and activator of transcriptio n 1/3 (STAT1/3), known as a transcription factor on COX-2 expression, is inactivated by simvastatin in both cells. Our findings showed that simvastatin is likely to suppress IFNgamma-induced COX-2 expression by inhibiting STAT1/3 activation in human THP-1 cells, but not in murine RAW264.7 cells. Thus, we concluded that IFNgamma-induced COX-2 expression is differently regulated by simvastatin depending on species specific mechanism.(AU)