γ-Sitosterol from Acacia nilotica L. induces G2/M cell cycle arrest and apoptosis through c-Myc suppression in MCF-7 and A549 cells

Ethnopharmacological relevance

Acacia nilotica is widely distributed in Asia. In India, it occupies an important place in the indigenous system of medicine against anti-inflammatory, antioxidant, cancers, and/or tumors.

Aim of the study

The purpose of this study is to investigate the inhibitory effect of Acacia nilotica leaves extract and γ-Sitosterol on cell proliferation, the apoptotic effect and cell cycle arrest in breast and lung cancer cells.

Materials and methods

GC–MS and HPLC were used to determine the chemical constituents of this extract and γ-Sitosterol respectively. Human MCF-7 and A549 cell lines were treated with Acacia nilotica extract and γ-Sitosterol. Cell viability was determined by MTT assay. Cell proliferation was determined by BrdU incorporation assay. Apoptosis was detected by cell morphologic observation through AO/EtBr staining, cell cycle analysis, and immunoblot analysis on the expression of protein associated with cell cycle arrest.

Results

Experimental results of bioactive compound analysis indicate that γ-Sitosterol, bioactive ingredients of Acacia nilotica extract. The IC₅₀ value of extract on MCF-7 and A549 cancer cells was 493.3 ± 15.2 and 696.6 ± 11.5 μg/ml, respectively. Acacia nilotica extract and γ-Sitosterol were inhibited the cell proliferation by 54.34 ± 1.8 and 42.18 ± 3.9% for MCF-7 and 58.26 ± 1.5 and 44.36 ± 3.05% for A549 cells. The percentage of apoptotic cells observed in the MCF-7 and A549 cell lines were increased to 42.46 and 36.8% of extract; 46.68 and 43.24% for γ-Sitosterol respectively. Flow cytometric analysis results demonstrate that cells were arrested at the G2/M phase and decrease the c-Myc expression.

Conclusions

This study demonstrates in vitro results, which support the ethnomedical use of γ-Sitosterol against cancer. Experimental results of this study suggest that γ-Sitosterol exerts potential anticancer activity through the growth inhibition, cell cycle arrest and the apoptosis on cancer cells.     

A Comprehensive Analysis of Plausible Genotoxic Covariates Among Workers of a Polyvinyl Chloride Plant Exposed to Vinyl Chloride Monomer

The aim of this study was to assess the frequency of chromosomal aberrations—including chromatid type aberrations (CTAs), chromosomal type aberrations, micronucleus (MN) comet assay, and XRCC1 399 Arg/Gln polymorphism—in peripheral blood lymphocytes of workers occupationally exposed to vinyl chloride monomer (VCM). A total of 52 workers and an equal number of controls were recruited into the study to explore the potential cytogenetic risk of occupational exposure to VCM. Questionnaires were administered to obtain details of habitual cigarette-smoking, alcohol-consumption pattern, and occupation, etc. The exposed subjects and controls were classified into two groups based on age (group I <40 years; group II ≥40 years), and exposed subjects were further classified based on exposure duration (>8 and ≥8 years). CTA, MN, and comet assay frequency were significantly greater in polyvinyl chloride (PVC) factory workers (p < 0.05) with long-duration work. CTA, MN, and comet assay values were found to be increased with age in exposed subjects as well as in controls, with exposed subjects showing a statistically greater degree. An extensively greater MN frequency was observed in smokers exposed to VCM than in the control group (P < 0.05). The mean tail length of exposed subjects was greater compared with controls. The study on XRCC1 399 Arg/gln polymorphism in PVC factory workers showed less significant difference in allele frequency compared with controls. In conclusion, this results of work provides evidence for an apparent genotoxic effect associated with VCM exposure. Our results reinforce the greater sensitivity of cytogenetic assays for biomonitoring of occupationally exposed populations. Statistics indicate that workers exposed to VCM are at carcinogenic risk and should be monitored for long-term adverse effects from their exposure.     

Bioactive compounds extracted from Indian wild legume seeds: antioxidant and type II diabetes-related enzyme inhibition properties

Seven different wild legume seeds (Acacia leucophloea, Bauhinia variegata, Canavalia gladiata, Entada scandens, Mucuna pruriens, Sesbania bispinosa and Tamarindus indica) from various parts of India were analyzed for total free phenolics, l-Dopa (l-3,4 dihydroxyphenylalanine), phytic acid and their antioxidant capacity (ferric-reducing antioxidant power [FRAP] and 2,2-diphenyl-1-picrylhydrazyl [DPPH] assay) and type II diabetes-related enzyme inhibition activitiy (α-amylase). S. bispinosa had the highest content in both total free phenolics and l-Dopa, and relatively low phytic acid when compared with other seeds. Phytic acid content, being highest in E. scandens, M. pruriens and T. indica, was highly predictive for FRAP (r = 0.47, p < 0.05) and DPPH (r = 0.66, p < 0.001) assays. The phenolic extract from T. indica and l-Dopa extract from E. scandens showed significantly higher FRAP values among others. All seed extracts demonstrated a remarkable reducing power (7-145 mM FeSO4 per mg extract), DPPH radical scavenging activity (16-95%) and α-amylase enzyme inhibition activity (28-40%).     

Analysis of Genetic Damage and Gene Polymorphism in Hepatocellular Carcinoma (HCC) Patients in a South Indian Population

Background

Hepatocellular carcinoma (HCC) is the second leading cause of cancer death in many regions of Asia and the etiology of human HCC is clearly multi-factorial. The development of effective markers for the detection of HCC could have an impact on cancer mortality and significant health implications worldwide. The subjects presented here were recruited based on the serum alpha-fetoprotein level, which is an effective marker for HCC. Further, the chromosomal alterations were elucidated using trypsin G-banding. HCCs with p53 mutations have high malignant potential and are used as an indicator for the biological behavior of recurrent HCCs. The functional polymorphism in the XRCC1 gene, which participates in the base-excision repair of oxidative DNA damage, was associated with increased risk of early onset HCC. Thus, in this investigation, the p53 and XRCC1 gene polymorphisms using the standard protocols were also assessed to find out whether these genes may be associated with HCC susceptibility.

Methods

Blood samples from HCC patients (n = 93) were collected from oncology clinics in South India. Control subjects (n = 93) who had no history of tumors were selected and they were matched to cases on sex, age, and race. Peripheral blood was analyzed for chromosomal aberrations (CAs) and micronuclei (MN) formation. p53 and XRCC1 genotypes were detected using a PCR–RFLP technique.

Results

Specific biomarkers on cytogenetic endpoints might help in diagnosis and treatment measures. The frequencies of genotypes between groups were calculated by χ² test. A statistically significant (p < 0.05) increase in CA was observed in HCC patients compared to their controls as confirmed by ANOVA and MN shows insignificant results. The study on p53 Arg72Pro and XRCC1 Arg399Gln polymorphism in HCC patients demonstrated differences in allele frequencies compared to their controls.

Conclusions

The present study indicates that chromosomal alterations and the genetic variations of p53 and XRCC1 may contribute to inter-individual susceptibility to HCC. A very limited role of genetic polymorphism was investigated in modulating the HCC risk, but the combined effect of these variants may interact to increase the risk of HCC in the South Indian population.     

Cytotoxic Effect of Palladium Nanoparticles Synthesized From <Emphasis Type="Italic">Syzygium</Emphasis><Emphasis Type="Italic">aromaticum</Emphasis> Aqueous Extracts and Induction of Apoptosis in Cervical Carcinoma

Among the various nanoparticles explored for diverse biological applications, palladium nanoparticles still remain far too behind in the field of cancer therapeutics. Nanoparticles synthesized by chemical methods aggregate in physiological conditions hindering their biomedical applications. Therefore, in the present study, a plant mediated green synthesis approach for palladium nanoparticle preparation from Syzygium aromaticum was reported and the biocompatibility and anticancer activity of the eco-friendly synthesized palladium nanoparticles against human cervical carcinoma was evaluated. The as-synthesized palladium nanoparticles were characterized by various analytical techniques such as, UV–vis spectroscopy, transmission electron microscope, dynamic light scattering, zeta potential, energy dispersive X-ray spectroscopy, X-ray diffraction and fourier transform infrared spectroscopy. The biocompatibility of palladium nanoparticles were verified by incubating with RBCs and cytotoxic studies revealed a dose dependent cytotoxic effect with half maximal inhibitory concentration values of 15 ± 0.5 µg/mL against HeLa cells at 48 h incubation. Further, the induction of apoptosis was evidenced by fluorescence microscopic study. Semi-quantitative RT-PCR analysis evidenced the activation of cytochrome c and caspase 3 and down regulation of Bcl-2 and Bcl-xL. In vivo antitumor studies showed significant suppression of tumor growth against HeLa tumor xenograft models. The results suggest that palladium nanoparticles can be synthesized using clove buds; they are biocompatible possessing significant anticancer activity against human cervical carcinoma, indicating the great potential of palladium nanoparticles in relevant biomedical applications.     

Health benefits of nut consumption with special reference to body weight control

Nuts are an integral part of the Mediterranean food patterns, and their incorporation into the regular diets of human beings is believed to provide many health benefits. The recent recognition of nuts as “heart-healthy” foods by the U.S. Food and Drug Administration has given a major boost to the positive image of nuts. Nut consumption has been associated with several health benefits, such as antioxidant, hypocholesterolemic, cardioprotective, anticancer, anti-inflammatory, and antidiabetic benefits, among other functional properties. However, although nuts possess these many health benefits, their consumption has been hampered by a lack of adequate information regarding those benefits. In addition, because nuts are energy-dense foods with high-fat content, there is a misconception among consumers that increased consumption may lead to unwanted gain in body weight with the risk of developing overweight/obesity. Nonetheless, available epidemiologic studies and short-term controlled feeding trials have supported the theory that the inclusion of nuts in the typical diet does not induce weight gain, despite an expected increase in total caloric intake. To address the misperception about nuts and body weight gain, the present review focuses mainly on the relation between nut consumption and body weight gain, in the context of the many health benefits of nuts.     

Identification and comparison of chromosomal alterations in infertile and fertile males of Tamil Nadu region exposed to cigarette smoking

Aim  

The aim of this investigation was to identify whether cigarette smoke increases the frequency of chromosomal aberrations (CA) in male infertility patients. Our study also endeavored to analyze the Y-chromosome deletion in infertile subjects.     

Assessment of Occupational Cytogenetic Risk, Among Petrol Station Workers

The focal aim of this study was to assess the frequency of chromosomal aberrations (CA) including chromatid type aberrations (CTA) and chromosomal type aberrations (CSA), micronucleus (MN) and XRCC1 399 Arg/Gln polymorphism in the peripheral blood lymphocytes of 27 petrol pump workers and same number of controls to explore the possible cytogenetic risk on occupational exposure to petrol vapors. The exposed subjects and controls were classified into two groups based on their age (group I < 40 years; group II > 40 years) apart from the classification of the exposed subjects based on their exposure duration (> 8 and < 8 years). CTA and MN frequency were significantly higher in petrol pump workers (p < 0.05) with longer work duration. CTA was found to increase with age in the exposed subjects as well as controls, with exposed subjects showing a statistically higher degree. This effect was not observed in MN. A significantly higher frequency of MN was observed in the smoking petrol pump workers than in control smokers (p < 0.05). No association was found between smoking and CA in both subjects. The study on XRCC1 399 Arg/gln polymorphism in petrol pump workers demonstrated very less difference in allele frequency compared to controls. In conclusion, these datas indicate that petrol pump workers under risk group should be monitored for any long-term adverse effects of the exposure.     

Flavonoid content in ethanolic extracts of selected raw and traditionally processed indigenous foods consumed by vulnerable groups of Kenya: antioxidant and type II diabetes-related functional properties

The present study evaluated the flavonoid content, antioxidant as well as type II diabetes-related enzyme inhibition activities of ethanolic extract of certain raw and traditionally processed indigenous food ingredients including cereals, legumes, oil seeds, tubers, vegetables and leafy vegetables, which are commonly consumed by vulnerable groups in Kenya. The vegetables exhibited higher flavonoid content (50-703?mg/100?g) when compared with the grains (47-343?mg/100?g). The ethanolic extract of presently studied food ingredients revealed 33-93% DPPH radical scavenging capacity, 486-6,389?mmol Fe(II)/g reducing power, 19-43% α-amylase inhibition activity and 14-68% α-glucosidase inhibition activity. Among the different food-stuffs, the drumstick and amaranth leaves exhibited significantly higher flavonoid content with excellent functional properties. Roasting of grains and cooking of vegetables were found to be suitable processing methods in preserving the functional properties. Hence, such viable processing techniques for respective food samples will be considered in the formulation of functional supplementary foods for vulnerable groups in Kenya.