Molecular biomarkers NOTCH1, CD44, BMI1, and TP53 in oral squamous cell carcinoma.

It is of interest to evaluate NOTCH1, CD44, BMI1, and TP53 genes in the epiglottis, tongue, and hard palate of oral malignancies (OM) with healthy controls. This was a prospective and cross-sectional study of 60 individuals with oral malignancies (OM) (20 each of tongue, epiglottis, and hard palate) studied at Malla Reddy Medical College and tertiary care hospitals in Hyderabad. Adults aged ≥ 18 years and diagnosed with oral cancer were included in the study. Those who had cancer in more than one area were excluded from the study. Blood samples of individuals with tongue or epiglottis or hard palate were taken for testing the expression of NOTCH1, CD44, TP53, and BMI1 genes. They were analysed by the genomic sequencing method. One-way ANOVA with Bonferroni's t-test was used for statistical analysis. Expression of NOTCH1, CD44, BMI1, and TP53 genes were significantly higher in epiglottis, tongue, and hard palate compared to healthy control samples (p < 0.001). All four genes were expressed in all three areas of OM. However, they were not significant between them. Further analysis revealed that NOTCH1, CD44, TP53, and BMI1 genes did not show any difference in HPV-positive and HPV-negative samples. Comparing the T stages of cancer Notch1, gene expression was significantly higher in stages 1 and 2 compared to 3 and 4. The CD44, TP53, and BMI1 did not show any differences in the T stage. However, the difference in HPV in all T stages was very minimal. Data showed that irrespective of the areas of cancer (epiglottis, tongue, and hard palate) NOTCH1, CD44, TP53, and BMI1 genes were expressed equally. The expression was not very much dependent on HPV positive (+ve) or negative (-ve). However the T-stage was showing higher expression compared to control group. Since the expression of these genes was very high in all the three malignancies, they may be used as early biomarkers to detect cancer of epiglottis, tongue, and hard palate.

Aqueous extract of Tamarindus indica fruit pulp exhibits antihyperglycaemic activity.

OBJECTIVE: Tamarindus indica Linn. (T.indica) is a well-known plant used in traditional medicine. The plant is popular for its antidiabetic activity. However, effect so f its aqueous fruit pulp extract on carbohydrate hydrolyzing enzymes and its glucose uptake potential were not explored. MATERIALS AND METHODS: The antidiabetic activity was assessed by in-vitro α-amylase and α-glucosidase inhibitory assays after preliminary phytochemical analysis. MTT assay was carried out to find cytotoxicity. Glucose uptake activity of the extract was carried out using L6 myotubes. RESULTS: The results showed a strong α-amylase inhibitory activity for the fruit pulp extract of T.indica compared to standard acarbose; the IC50 of the fruit pulp extract of T.indica and acarbose was 34.19 µg/ml 34.83µM. The extract also showed moderate α-glucosidase inhibitory activity. IC50 of the fruit pulp extract of T.indica and acarbose were 56.91µg/ml and 45.69µM respectively. The cytotoxicity assay showed IC50 of >300µg/ml and ≥1000µM for the fruit pulp extract of T.indica and metformin. The extract showed 63.99±0.08% glucose uptake in L6 myotubes whereas metformin and insulin at 10µg/ml and 10µM exhibited an uptake of 76.99±0.3% and 84.48±0.45% glucose, respectively. CONCLUSION: The study revealed that the fruit pulp extract of T.indica Linn does not show any cytotoxic effect and has very good α-amylase and good α-glucosidase inhibitory activities. The glucose uptake potential proves its postprandial hypoglycemic effect. Hence, it may be considered an antidiabetic agent for control of postprandial hyperglycemia.

Antidiabetic Activity of Ajwain Oil in Different In Vitro Models.

CONTEXT: Ajwain oil is an essential oil with thymol as its major constituent which is known for many pharmacological activities. AIMS: To evaluate the anti-hyperglycaemic potential of ajwain oil using different in vitro models. METHODS AND MATERIAL: In vitro α-amylase and α-glucosidase inhibitory effect of ajwain oil was carried out by the method of Bernfeld and Shibano et al. 1997 with minor modifications respectively. Cytotoxicity of the ajwain oil was assessed using MTT assay. Glucose uptake potential was assessed in differentiated L6 myotubes using fluorescent tagged 6-NBDG. RESULTS: Ajwain oil showed very good α - amylase inhibitory activity. A maximum inhibition of 88.55 ±0.43 % was achieved at a concentration of 4µL/ml by ajwain oil which was comparable to that of standard acarbose, 90.96 ± 1.81%. The IC50 of the extract was found to be 0.47µL/ml and for acarbose 0.69µL/ml. The maximum in vitro α - glucosidase inhibitory activity was found to be 89 ± 0.72 % and 91.67 ± 1.09% at 4µL/ml for ajwain oil and acarbose. The IC50 of the extract and acarbose were found to be 0.37µL/ml and 0.41µL/ml respectively. Ajwain oil has enhanced glucose uptake in L6 myotubes in a dose dependent manner. CONCLUSION: The anti-hyperglycaemic activity of the ajwain oil strongly support its ability to decrease sugar level hence it may be further validated for its use as an antidiabetic agent.

Oxidative stress and neuromodulatory effects of deltamethrin and its combination with insect repellents in rats.

Deltamethrin (DEL) and other synthetic repellents like N,N-diethyl benzamide (DEB), N,N-diethyl phenylacetamide (DEPA), and N,N-diethyl meta toluamide (DEET) are widely used due their high efficacy against mosquitoes. We evaluated the safe dose on exposure to insecticide and insect repellent individually and in combination in Wistar rats. Rats were administrated with individual chemicals DEL, DEB, DEPA, and DEET and in combination of DEL along DEB, DEPA, and DEET. The reduced glutathione and its detoxifying enzymes were significantly increased in DEL treated rats. Serotonin, dopamine, and noradrenaline were increased in DEL alone and its combination groups. Histopathology revealed that DEL and along with other insect repellants can cause lung toxicities. This study suggests that DEL in combination with other insect repellents showed antagonistic effect against oxidative stress, and there was no significant toxicological effect in the combination groups. The insect repellents with DEL cause less toxic effect and is safe for use.

Antihyperglycemic Activity of Caralluma fimbriata: An In vitro Approach.

BACKGROUND: An increase in prevalence of diabetes mellitus necessitates the need to develop new drugs for its effective management. Plants and their bioactive compounds are found to be an alternative therapeutic approach. Caralluma fimbriata, used in this study, is well known for its various biological effects. OBJECTIVE: The present study was designed to investigate the antihyperglycemic effect of the ethanolic leaf extract of C. fimbriata. MATERIALS AND METHODS: Different concentrations (1-1000 mg/mL) of the ethanolic leaf extract of C. fimbriata were subjected to alpha-amylase and alpha-glucosidase inhibitory assay with acarbose as control. Cytotoxicity was assessed by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Glucose uptake assay was performed on L6 myotubes using the extract in 1 µg-100 µg/mL, using metformin and insulin as control. RESULTS: The C. fimbriata extract showed potent inhibitory activity on enzymes of glucose metabolism in a dose-dependent manner. The maximum alpha-amylase inhibitory effect was 77.37% ± 3.23% at 1000 µg/mL with an IC50 value of 41.75 µg/mL and alpha-glucosidase inhibitory effect was 83.05% ± 1.69% at 1000 µg/mL with an IC50 value of 66.71 µg/mL. The maximum glucose uptake was found to be 66.32% ± 0.29% for the Caralluma extract at 100 µg/mL and that of metformin (10 µg/mL) was 74.44% ± 1.72% and insulin (10 mM) 85.55% ± 1.14%. The extract was found to be safe as the IC50 of extract and metformin was found to be ≥1000 µg/mL and ≥1000 µM, respectively, in the cell line tested. CONCLUSION: The study concludes that C. fimbriata has promising antihyperglycemic activity. SUMMARY: Caralluma fimbriata extract exhibited effective dose dependent inhibitory activity against alpha-amylase and alpha- glucosidaseEnhanced glucose uptake from L6 myotubes was appreciated in the presence of the extract, comparable to Insulin and metforminCaralluma fimbriata has potent antihyperglycemic properties. Abbreviations used: GLUT: Glucose transporter; MTT: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.