Nripendranath Mandal

Antioxidant and free radical scavenging activity of Spondias pinnata

BackgroundMany diseases are associated with oxidative stress caused by free radicals. Current research is directed towards finding naturally-occurring antioxidants of plant origin. The aim of the present study was to evaluate the in vitro antioxidant activities of Spondias pinnata stem bark extract.MethodsA 70% methanol extract of Spondias pinnata stem bark was studied in vitro for total antioxidant activity, for scavenging of hydroxyl radicals, superoxide anions, nitric oxide, hydrogen peroxide, peroxynitrite, singlet oxygen and hypochlorous acid, and for iron chelating capacity, reducing power, and phenolic and flavonoid contents.ResultsThe extract showed total antioxidant activity with a trolox equivalent antioxidant concentration (TEAC) value of 0.78 ± 0.02. The IC50 values for scavenging of free radicals were 112.18 ± 3.27 μg/ml, 13.46 ± 0.66 μg/ml and 24.48 ± 2.31 μg/ml for hydroxyl, superoxide and nitric oxide, respectively. The IC50 for hydrogen peroxide scavenging was 44.74 ± 25.61 mg/ml. For the peroxynitrite, singlet oxygen and hypochlorous acid scavenging activities the IC50 values were 716.32 ± 32.25 μg/ml, 58.07 ± 5.36 μg/ml and 127.99 ± 6.26 μg/ml, respectively. The extract was found to be a potent iron chelator with IC50 = 66.54 ± 0.84 μg/ml. The reducing power was increased with increasing amounts of extract. The plant extract (100 mg) yielded 91.47 ± 0.004 mg/ml gallic acid-equivalent phenolic content and 350.5 ± 0.004 mg/ml quercetin-equivalent flavonoid content.ConclusionThe present study provides evidence that a 70% methanol extract of Spondias pinnata stem bark is a potential source of natural antioxidants.

Assessment of the Antioxidant and Reactive Oxygen Species Scavenging Activity of Methanolic Extract of Caesalpinia crista Leaf

“Oxidative stress” is initiated by reactive oxygen species (ROS), which are responsible for majority of the diseases. However, antioxidants with ROS scavenging ability may have great relevance in the prevention of oxidative stress. The present study was undertaken, using a 70% methanolic extract of Caesalpinia crista leaves, to examine different in vitro tests in diversified fields including total antioxidant activity, scavenging activities for various ROS, iron chelating activity and phenolic and flavonoid contents. Total antioxidant activity was evaluated as trolox equivalent antioxidant capacity value of 0.546 ± 0.014. The extract was investigated for different ROS scavenging activities and IC50 values were found to be 0.44 ± 0.1 mg/ml, 24.9 ± 0.98 μg/ml, 33.72 ± 0.85 μg/ml, 61.13 ± 3.24 μg/mL and 170.51 ± 4.68 μg/mL for hydroxyl, superoxide, nitric oxide, singlet oxygen and hypochlorous acid, respectively; however, no significant results were obtained in scavenging of hydrogen peroxide and peroxynitrite anion. The extract was found to be a potent iron chelator with IC50 = 279.85 ± 4.72 μg/mL. The plant extract (100 mg) yielded 50.23 ± 0.003 mg/mL gallic acid equivalent phenolic content and 106.83 ± 0.0003 mg/mL quercetin equivalent flavonoid content. In the in vivo experiments, the extract treatment showed significant increase in the level of superoxide dismutase, catalase, glutathione-S-transferase and reduced glutathione. In a word, it may be concluded that 70% methanol extract of C. crista leaves acts as an antioxidant and ROS scavenger; which may be due to the presence of phenolic and flavonoid compounds.

Relation of Anti- to Pro-Inflammatory Cytokine Ratios with Acute Myocardial Infarction

Background/Aims Acute myocardial infarction (AMI) is a leading cause of death. Inflammatory processes play an important role in atherosclerosis, which is intimately related to AMI. The aim of this study was to investigate the association between anti-inflammatory and pro-inflammatory cytokines ratios and AMI. Methods A total of 90 AMI patients and 90 age-and sex-matched controls were recruited in this study. Plasma cytokines and conventional risk factors were determined by standard methods. Results Patients with AMI showed increased interleukin (IL)-6 and tumor necrosis factor-α levels and lower anti- to pro-inflammatory cytokine ratios as compared with controls. A multivariate logistic regression analysis revealed that IL-10 to IL-6 ratio was independently associated with the occurrence of AMI (odds ratio [OR], 5.39; 95% confidence interval [CI], 2.39 to 12.17; p < 0.0001). In contrast, IL-6 levels were no longer significant in the multivariate model (OR, 1.02; 95% CI, 0.932 to 1.12; p = 0.603). A receiver operating characteristic (ROC) curve analysis indicated that IL-6 levels and IL-10 to IL-6 ratios were a significant predictor of AMI (area under ROC curve, 0.892 and 0.851, respectively). Conclusions Our results suggest that the ratio of IL-10 to IL-6 is independently associated with AMI, and reduced levels of this ratio may favor the development of AMI.

An antioxidant extract of tropical lichen, Parmotrema reticulatum, induces cell cycle arrest and apoptosis in breast carcinoma cell line MCF-7.

This report highlights the phytochemical analysis, antioxidant potential and anticancer activity against breast carcinoma of 70% methanolic extract of lichen, Parmotrema reticulatum (PRME). Phytochemical analysis of PRME confirms the presence of various phytoconstituents like alkaloids, carbohydrates, flavonoids, glycosides, phenols, saponins, tannins, anthraquinones, and ascorbic acid; among which alkaloids, phenols and flavonoids are found in abundant amount. High performance liquid chromatography (HPLC) analysis of PRME revealed the presence of catechin, purpurin, tannic acid and reserpine. Antioxidant activity was evaluated by nine separate methods. PRME showed excellent hydroxyl and hypochlorous radical scavenging as well as moderate DPPH, superoxide, singlet oxygen, nitric oxide and peroxynitrite scavenging activity. Cytotoxicity of PRME was tested against breast carcinoma (MCF-7), lung carcinoma (A549) and normal lung fibroblast (WI-38) using WST-1 method. PRME was found cytotoxic against MCF-7 cells with an IC50 value 130.03±3.11 µg/ml while negligible cytotoxicity was observed on A549 and WI-38 cells. Further flow cytometric study showed that PRME halted the MCF-7 cells in S and G2/M phases and induces apoptosis in dose as well as time dependent manner. Cell cycle arrest was associated with downregulation of cyclin B1, Cdk-2 and Cdc25C as well as slight decrease in the expression of Cdk-1 and cyclin A1 with subsequent upregulation of p53 and p21. Moreover PRME induced Bax and inhibited Bcl-2 expression, which results in increasing Bax/Bcl-2 ratio and activation of caspase cascade. This ultimately leads to PARP degradation and induces apoptosis in MCF-7 cells. It can be hypothesised from the current study that the antioxidant and anticancer potential of the PRME may reside in the phytoconstitutents present in it and therefore, PRME may be used as a possible source of natural antioxidant that may be developed to an anticancer agent.

Synergistic effect of anti and pro-inflammatory cytokine genes and their promoter polymorphism with ST-elevation of myocardial infarction.

BACKGROUND The single-gene approach in association studies of polygenic diseases such as acute myocardial infarction (AMI) is likely to provide limited value. Interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) plasma levels may be genetically influenced. AIM We evaluate the impact of single nucleotide polymorphism of the promoter region of these genes, as well as reciprocal interaction of these genes with ST-elevation of myocardial infarction (STEMI). METHODS In a case-control study 500 STEMI patients and 500 age- and sex-matched controls were studied. Three single-nucleotide polymorphism genotypes were evaluated by polymerase chain reaction and restriction enzyme analysis and assessed their association with STEMI. The synergistic effects of IL-6, TNF-α and IL-10 gene polymorphisms were evaluated by using logistic regression analysis. RESULTS We found that IL-6 and TNF-α concentrations of studied population were significantly different (p<0.0001) in each genotype of IL-6 -174G>C and TNF-α -308G>A gene polymorphisms respectively. A significant association was found in multivariate analysis for the IL-6 -174G>C [odds ratio (OR): 0.390; 95% confidence interval (CI): 0.176-0.865, p=0.020] and TNF-α -308G>A [OR: 0.372; 95% CI: 0.171-808, p=0.012] gene polymorphisms with STEMI. In contrast, IL-10 -592C>A gene polymorphism was no longer significant in the multivariate model (OR: 0.678; 95% CI: 0.288 to 1.594, p=0.373) whereas significant in univariate analysis (OR: 0.697; 95% CI: 0.523-0.929, p=0.014). CONCLUSIONS Our findings suggest that IL-6, TNF-α and IL-10 gene polymorphisms all contribute in the association with STEMI whereas the association persisted only for IL-6 and TNF-α but not for IL-10 gene polymorphism with this disease in the multivariate analysis.

Reducing power and iron chelating property of Terminalia chebula (Retz.) alleviates iron induced liver toxicity in mice

BackgroundThe 70% methanol extract of Terminalia chebula Retz. fruit (TCME) was investigated for its in vitro iron chelating property and in vivo ameliorating effect on hepatic injury of iron overloaded mice.MethodsThe effect of fruit extract on Fe2+-ferrozine complex formation and Fe2+ mediated pUC-18 DNA breakdown was studied in order to find the in vitro iron chelating activity. Thirty-six Swiss Albino mice were divided into six groups of: blank, patient control and treated with 50, 100, 200 mg/kg b.w. of TCME and desirox (standard iron chelator drug with Deferasirox as parent compound). Evaluations were made for serum markers of hepatic damage, antioxidant enzyme, lipid per oxidation and liver fibrosis levels. The reductive release of ferritin iron by the extract was further studied.ResultsIn vitro results showed considerable iron chelation with IC50 of 27.19 ± 2.80 μg/ml, and a significant DNA protection with [P]50 of 1.07 ± 0.03 μg/ml along with about 86% retention of supercoiled DNA. Iron-dextran injection (i.p.) caused significant increase in the levels of the serum enzymes, viz., alanine aminotransferase (ALAT), aspartate aminotransferase (ASAT), alkaline phosphatase (ALP) and Bilirubin, which were subsequently lowered by oral administration of 200 mg/kg b.w. dose of the fruit extract by 81.5%, 105.88%, 188.08% and 128.31%, respectively. Similarly, treatment with the same dose of the extract was shown to alleviate the reduced levels of liver antioxidant enzyme superoxide dismutase, catalase, glutathione S-transferase and non-enzymatic reduced glutathione, by 49.8%, 53.5%, 35.4% and 11% respectively, in comparison to the iron overloaded mice. At the same time, the fruit extract effectively lowered the iron-overload induced raised levels of lipid per oxidation, protein carbonyl, hydroxyproline and liver iron by 49%, 67%, 67% and 26%, respectively, with oral treatment of 200 mg/kg b.w. dose of TCME. The fruit extract also showed potential activity for reductive release of ferritin iron.ConclusionsThese findings suggest that Terminalia chebula extract may contain active substances capable of lessening iron overload induced toxicity, and hence possibly be useful as iron chelating drug for iron overload diseases.

Assessment of in Vitro Antioxidant and Free Radical Scavenging Activity of Cajanus cajan

The present study was undertaken to evaluate the antioxidant and free radical scavenging activities of 70% methanolic extract of Cajanus cajan leaves using various in vitro assays. The TEAC value 0.84 ± 0.05 for the extract indicates it is an antioxidant. The extract had shown its scavenging activity for different free radicals and 175.73 ± 7.50 ?g/ml, 27.17 ± 0.9 ?g/ml, 60.78 ± 5.15 ?g/ml and 250.29 ± 5.70 ?g/ml were determined as the IC50 values for hydroxyl, superoxide, nitric oxide and singlet oxygen radicals, respectively. The sample showed reasonable chelation of Fe2+ ion and inhibition of lipid peroxidation. The phenolic and flavonoid contents of the extract were also determined to be 51.33 ± 0.002 mg/ml gallic acid equivalent and 143.0 ± 0.003 mg/ml quercetin equivalent, respectively. The results showed that 70% methanol extract of C. cajan leaves possesses antioxidant and free radical scavenging properties.

Self-Assembled Novel BODIPY-Based Palladium Supramolecules and Their Cellular Localization

Four new palladium metal supramolecules with triangular/square architectures derived from boron dipyrromethane (BODIPY) ligands were synthesized by self-assembly and fully characterized by 1H and 31P NMR, electrospray ionization mass spectrometry, and single-crystal X-ray diffraction. These supramolecules were more cytotoxic to brain cancer (glioblastoma) cells than to normal lung fibroblasts. Their cytotoxicity to the glioblastoma cells was higher than that of a benchmark metal-based chemotherapy drug, cisplatin. The characteristic green fluorescence of the BODIPY ligands in these supramolecules permitted their intracellular visualization using confocal microscopy, and the compounds were localized in the cytoplasm and on the plasma membrane.

Heartwood extract of Acacia catechu induces apoptosis in human breast carcinoma by altering bax/bcl-2 ratio

Background: The heartwood extract of A. catechu, called pale catechu or “Katha” in Hindi has been widely used in traditional Indian medicinal system. Although various pharmacological properties of this plant had been reported previously, only a few were concerned with the anticancer activity of this plant. Objective: The objective was to assess the in vitro anticancer and apoptosis inducing effect of 70% methanolic extract of “Katha” (ACME) on human breast adenocarcinoma cell line (MCF-7). Materials and Methods: MCF-7 cell line was treated with increasing concentrations of ACME and cell viability was calculated. Flow cytometric methods were used to confirm the apoptosis promoting role of ACME. Morphological changes were then analysed using confocal microscopy. Western blotting was then performed to investigate the expression of apoptogenic proteins and to analyse the activation of caspases. Results: ACME showed significant cytotoxicity to MCF-7 cells with an IC50 value of 288.85 ± 25.79 μg/ml. Flow cytometric analysis and morphological studies confirmed that ACME is able to induce apoptosis in MCF-7 cells. Furthermore, immunoblot results suggested the pathway of apoptosis induction by increasing Bax/Bcl-2 ratio which results in the activation of caspase-cascade and ultimately leads to the cleavage of Poly adeno ribose polymerase (PARP). Conclusion: These results provide the evidence that ACME is able to inhibit the proliferation of MCF-7 cells by inducing apoptosis through intrinsic pathway.

Hepatoprotective Potential of Caesalpinia crista against Iron-Overload-Induced Liver Toxicity in Mice.

The present study was carried out to evaluate the ameliorating effect of Caesalpinia crista Linn. (CCME) extract on iron-overload-induced liver injury. Iron overload was induced by intraperitoneal administration of iron dextran into mice. CCME attenuated the percentage increase in liver iron and serum ferritin levels when compared to control group. CCME also showed a dose-dependent inhibition of lipid peroxidation, protein oxidation, and liver fibrosis. The serum enzyme markers were found to be less, whereas enhanced levels of liver antioxidant enzymes were detected in CCME-treated group. In presence of CCME, the reductive release of ferritin iron was increased significantly. Furthermore, CCME exhibited DPPH radical scavenging and protection against Fe2+-mediated oxidative DNA damage. The current study confirmed the hepatoprotective effect of CCME against the model hepatotoxicant iron overload and the activity is likely related to its potent antioxidant and iron-chelating property.