Alarm Test: A Novel Chemical-Free Behavioural Assessment Tool for Zebrafish

Zebrafish (ZF) is an incredible animal for the study of neurological disorders. Its behaviour is like higher vertebrate animals, which makes it gainful and robust. Understanding the psychological and biological implications of housing settings for ZFs is very crucial in improving the replicability and dependability of ZF behavioural research. Individual housing triggers depression-like symptoms that suggest that housing conditions have negative effects on ZF and can result in the data discrepancy. Based on various behavioural analyses, we have evaluated that the ZFs kept in isolation and the ZFs kept in herd conditions exhibit different behavioural patterns. Interestingly, normal isolated subjects exhibit similar behavioural patterns as Alzheimer disease (AD)-induced subjects; hence, this can have serious implications on any study concerning behaviour of ZFs. Therefore, we have reported a new behavioural test named “Alarm Test”, which effectively discriminates normal isolated subjects from AD subjects. Alarm Test is observed to be better than other tests used for studying fear and anxiety in ZFs as it uses the indigenous compound released by ZFs during fear and makes use of the same for analysis. This can reduce the involvement of chemicals during behavioural analysis as well as sacrifice of ZFs for collection of alarm substance

Molecular Mechanisms behind Initiation of Focal Seizure in Temporal Lobe Epilepsy: Computational Study

Epilepsy is a noncommunicable disease of the brain that affects people of all ages. The chapter aims to identify protein targets and their mechanism of action behind temporal lobe epilepsy. Differentially expressed proteins in temporal lobe epilepsy (TLE) were used to derive a hypothesis demonstrating routes of protein interactions causing focal seizure and identification of putative target receptor for its treatment. Text mining was done by constructing a Boolean query with keywords such as temporal lobe epilepsy, focal seizures, proteomics, etc., in different scientific search engines. The proteins were further used for creating protein interaction network and analysed for their role in focal epileptic seizure pathway. The most appropriate route for initiation of seizure was observed to be route 3. It describes the dysregulated signal transduction from adenosine A1 receptor (ADORA1) to gamma-aminobutyric acid (GABA) B receptor 1 (GABBR1). This causes electrical imbalance and hyper-excitation of neurons that lead to focal seizure. The study also predicts that YWHAZ (3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta) could be the potential target for preventing focal seizures. The network framed in this study is ideal for studying the cascades of events that may occur during focal seizures in TLE and is useful in drug discovery

SUSTAINED RELEASE TABLETS OF SORAFENIB-SILIBININ COMBINATIONS FOR THE TREATMENT OF HEPATOCELLULAR CARCINOMA

Objective: The aim of this study was to develop polymer coated sustained release tablet using sorafenib and silibinin combination for the treatment of hepatocellular carcinoma. Methods: The qualitative analysis such as weight variation, friability, hardness, interaction studies, disintegration and in vitro release were performed to validate formulated tablets. We have maintained the acceptable official limits for weight variation, friability, hardness and disintegration time according to prescribed pharmacopoeial recommendation. In vitro drug release studies were performed using USP-II (paddle type) dissolution apparatus. The MTT assay was performed for assessment of Cell viability of drug combination for tablet formulation. Molecular docking studies have been performed to determine the combinatorial mode of action for the tablet formulation. Results: Friability and weight variation were less than 1% for each formulation, which were within range of prescribed pharmacopoeial recommendation. The hardness of 20 tablets showed 5-6.5Kg/cm2 for all formulations 5-6.5Kg/cm2. The optimized formulation resulted in 98% drug release after 28 h. The present study reports the synergistic effects of drug combination to inhibit cell growth in HepG2 cell line. Molecular docking studies showed that sorafenib has high binding affinity for B-Raf vascular endothelial growth factor receptor β and protein kinase B. Silibinin showed binding affinity with MAP kinase-11, protein phosphatase 2 A and tankyrase. Conclusion: The present study reports for the first time a novel formulation for sustained release and reduced toxicity of sorafenib with enhanced inhibitory effect of the drug combination on cancerous hepatic cell line as well collaborative mechanism of action for the formulation

Mucin 13 expression correlates with tumor development in hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of early diagnostic marker. Accumulating studies have shown that elevated expression of mucin 13 as potential oncogene and predictive biomarker for various cancer. However, very little is known about its expression and function for development and progression of HCC Objective: To investigate mucin 13 expression in chemically induced hepatocellular carcinoma model. Methodology: Diethyl nitrosamine (DEN) and 2-Acetylaminofluorene (2-AAF) induced method was employed for the development of hepatocellular carcinoma in Male Wistar rats. Serum and tissues were collected at regular interval of time and routinely validated for liver cancer stages. Immunohistochemistry and in situ hybridization were performed on formalin-fixed, paraffin-embedded tissues. Molecular docking studies were performed to study the interaction of mucin 13 and DEN. Results: Our results demonstrate hepatocellular adenoma as observed by histopathological analysis. Biochemical analysis showed a progressive increase in the levels of serum ALT, AST and ALP, suggesting the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually elevated during consecutive stages of hepatocellular carcinoma. Interestingly, an increase in nuclear localization of mucin 13 was observed in treated group as compared to control group. In situ hybridization analysis showed that a decrease in miR-132 and miR-145, which are inversely related with mucin 13 expression. Moreover, DEN efficiently binds mucin 13 with high affinity and thus stabilize it as demonstrated by molecular docking analysis. Conclusion: These results suggest that mucin 13 expression is closely associated with hepatocarcinogenesis and could serve as a predictive candidate biomarker for HCC

Mucin 13 expression is an early indicator of hepatocellular carcinoma development

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of an early marker for diagnosis. Studies show that increased mucin 13 (MUC13) expression as a possible oncogene and predictive biomarker for various cancers has been shown. But its expression and role in the development of HCC is very little known. Objective: The aim of this study is to investigate the MUC13 expression in chemically induced hepatocellular carcinoma model. Methodology: Male Wistar rats were subjected to a DEN and 2-Acetylaminofluorene (2-AAF) induced method for the development of hepatocellular carcinoma. Serum and tissues were collected at regular intervals and routinely validated for various stages of liver cancer. On formalin-fixed, paraffin-embedded tissues, immunohistochemistry and in situ hybridization were performed. The molecular interaction of mucin 13 and DEN were also performed using in silico analysis. Results: Histopathological analysis of liver tissues revealed the development of hepatocellular carcinoma with successive stages in chemically induced model HCC. Moreover, biochemical analysis showed a progressive increase in serum ALT, AST, and ALP levels, indicating the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually increased during the progression of hepatocellular carcinoma. The treated group showed an increase in nuclear localization of mucin 13 as compared to the control group. In situ hybridization analysis revealed a reduction in miR-132 and miR145, both of which are inversely related to mucin 13 expression. Furthermore, molecular docking analysis showed that DEN efficiently binds mucin 13 with high affinity and thus stabilizes it. Conclusion: These findings suggest that mucin 13 expression is linked to hepatocarcinogenesis and could be used as a candidate biomarker for HCC

In silico approach to evaluate the efficacy of dietary flavonoids and their role in Alzheimer\u27s disease

© 2015, Global Research Online. All rights reserved. Alzheimer’s disease is recognized as fatal neurological disorder that leads to constant degeneration of neurons in later stage of life. Etiology of disease progression is characterized by accumulation of amyloid beta and tau protein that contributes in the formation of senile plaques and tangles. It was also observed that nutritional status of AD patients was low and often not considered in research studies. Here we emphasize on the intake of those dietary items which are good source of natural antioxidants and hence can protect the brain from oxidative and inflammatory damage. We have compared twenty seven secondary metabolites from 5 different classes of Flavonoids found in 506 dietary items (Source: USDA Database for Flavonoid Content of Selected foods, Release 3.1(2013)) along with marketed drugs for Alzheimer’s disease to AChE receptor. Our study indicates that Flavan-3-ol family of flavonoids are the best dietary supplements for improving brain health. Theaflavin-3’-gallate is the best binding ligand to AChE receptor. Pharmacophoric studies were conducted to see the potential of respective compounds as drug candidate. Since pharmaceutical industries have always shown interest in compounds from natural sources, a little intervention of these compounds with the synthetic drugs will guarantee lesser side effects with maximum efficacy. Hence the food items like hazelnuts, Green Tea, Black Tea, Pistachio nuts, walnuts, Cocoa mix, Coffee etc. should be incorporated more in the diet of healthy people as well as in AD patients to protect brain from damage and improve the cognitive abilities

Neuroprotective effect of sesame seed oil in 6-hydroxydopamine induced neurotoxicity in mice model: Cellular, biochemical and neurochemical evidence

Natural antioxidants have shown a remarkable reduction in oxidative stress due to excess formation of reactive oxygen species by enhancing antioxidant mechanism in the neurodegenerative disorders. Sesame seed oil (SO) is one of the most important edible oil in India as well as in Asian countries and has potent antioxidant properties thus the present study evaluated the neuroprotective effect of SO by using 6-Hydroxydopamine (6-OHDA)-induced Parkinson\u27s disease model in mice. The mice were fed an SO mix diet for 15 days and then 6-OHDA was injected into the right striatum of mice brain. Three weeks after 6-OHDA infusion, mice were sacrificed and the striatum was removed. The neuroprotective role of SO on the activities of antioxidant and non-antioxidant enzymes such as glutathione reductase (GR), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT) and content of glutathione (GSH) and thiobarbituric acid reactive substance (TBARS) were studied in the striatum. The activities of all the above-mentioned enzymes decreased significantly in 6-OHDA group (Lesioned) when compared with Sham. The pretreatment of SO on antioxidant mechanism and dopamine level in the brain had shown some significant improvement in Lesion+SO (L+SO) group when compared with Lesioned group. However, NADPH oxidase subunit, Nox2 and inflammatory stimulator Cox2 expression was increased as well as antioxidant MnSOD level was decreased in Lesioned group while SO showed the inhibitory effect on the activation of Nox2 and Cox2 and restored MnSOD expression in L+SO group. Increased tyrosine hydroxylase (TH) expression in substantia nigra as well as dopamine and its metabolite DOPAC level in L+SO group also support our findings that SO may inhibit activation of NADPH oxidase dependent inflammatory mechanism due to 6-OHDA induced neurotoxicity in mice. © Springer Science+Business Media, LLC 2011

Sesamin attenuates neurotoxicity in mouse model of ischemic brain stroke

© 2014 Elsevier Inc.. Stroke is a severe neurological disorder characterized by the abrupt loss of blood circulation into the brain resulting into wide ranging brain and behavior abnormalities. The present study was designed to evaluate molecular mechanism by which sesamin (SES) induces neuroprotection in mouse model of ischemic stroke. The results of this study demonstrate that SES treatment (30. mg/kg. bwt) significantly reduced infarction volume, lipid per-oxidation, cleaved-caspase-3 activation, and increased GSH activity following MCAO in adult male mouse. SES treatment also diminished iNOS and COX-2 protein expression, and significantly restored SOD activity and protein expression level in the ischemic cortex of the MCAO animals. Furthermore, SES treatment also significantly reduced inflammatory and oxidative stress markers including Iba1, Nox-2, Cox-2, peroxynitrite compared to placebo MCAO animals. Superoxide radical production, as studied by DHE staining method, was also significantly reduced in the ischemic cortex of SES treated compared to placebo MCAO animals. Likewise, downstream effects of superoxide free radicals i.e. MAPK/ERK and P38 activation was also significantly attenuated in SES treated compared to placebo MCAO animals. In conclusion, these results suggest that SES induces significant neuroprotection, by ameliorating many signaling pathways activated/deactivated following cerebral ischemia in adult mouse