Kanthi Kiran Kondepudi

Potential of probiotics, prebiotics and synbiotics for management of colorectal cancer.

Colorectal Cancer (CRC) is the second leading cause of cancer-related mortality and is the fourth most common malignant neoplasm in USA. Escaping apoptosis and cell mutation are the prime hallmarks of cancer. It is apparent that balancing the network between DNA damage and DNA repair is critical in preventing carcinogenesis. One-third of cancers might be prevented by nutritious healthy diet, maintaining healthy weight and physical activity. In this review, an attempt is made to abridge the role of carcinogen in colorectal cancer establishment and prognosis, where special attention has been paid to food-borne mutagens and functional role of beneficial human gut microbiome in evading cancer. Further the significance of tailor-made prebiotics, probiotics and synbiotics in cancer management by bio-antimutagenic and desmutagenic activity has been elaborated. Probiotic bacteria are live microorganisms that, when administered in adequate amounts, confer a healthy benefit on the host. Prebiotics are a selectively fermentable non-digestible oligosaccharide or ingredient that brings specific changes, both in the composition and/or activity of the gastrointestinal microflora, conferring health benefits. Synbiotics are a combination of probiotic bacteria and the growth promoting prebiotic ingredients that purport “synergism.”

Capsaicin Induces ''Brite'' Phenotype in Differentiating 3T3-L1 Preadipocytes

Objective Targeting the energy storing white adipose tissue (WAT) by pharmacological and dietary means in order to promote its conversion to energy expending “brite” cell type holds promise as an anti-obesity approach. Present study was designed to investigate/revisit the effect of capsaicin on adipogenic differentiation with special reference to induction of “brite” phenotype during differentiation of 3T3-L1 preadipocytes. Methods Multiple techniques such as Ca2+ influx assay, Oil Red-O staining, nutrigenomic analysis in preadipocytes and matured adipocytes have been employed to understand the effect of capsaicin at different doses. In addition to in-vitro experiments, in-vivo studies were carried out in high-fat diet (HFD) fed rats treated with resiniferatoxin (RTX) (a TRPV1 agonist) and in mice administered capsaicin. Results TRPV1 channels are expressed in preadipocytes but not in adipocytes. In preadipocytes, both capsaicin and RTX stimulate Ca2+ influx in dose-dependent manner. This stimulation may be prevented by capsazepine, a TRPV1 antagonist. At lower doses, capsaicin inhibits lipid accumulation and stimulates TRPV1 gene expression, while at higher doses it enhances accumulation of lipids and suppresses expression of its receptor. In doses of 0.1–100 µM, capsaicin promotes expression of major pro-adipogenic factor PPARγ and some of its downstream targets. In concentrations of 1 µM, capsaicin up-regulates anti-adipogenic genes. Low-dose capsaicin treatment of 3T3-L1 preadipocytes differentiating into adipocytes results in increased expression of brown fat cell marker genes. In white adipose of mice, capsaicin administration leads to increase in browning-specific genes. Global TRPV1 ablation (i.p. by RTX administration) leads to increase in locomotor activity with no change in body weight. Conclusion Our findings suggest the dual modulatory role of capsaicin in adipogenesis. Capsaicin inhibits adipogenesis in 3T3-L1 via TRPV1 activation and induces brown-like phenotype whereas higher doses.

Expression of multiple Transient Receptor Potential channel genes in murine 3T3-L1 cell lines and adipose tissue

BACKGROUND Calcium and its signaling have a role in adipogenesis. Transient Receptor Potential (TRP) channels are non-selective cation channels with a high permeability to calcium. METHODS In the present study the expression of multiple TRP channels on mouse 3T3-L1 preadipocyte and adipocyte cells, white (WAT) and brown (BAT) adipose tissues was investigated using real time PCR (RT-PCR). RESULTS TRPV1, TRPV3, TRPM8, TRPC4, TRPC6 were differentially expressed in preadipocytes and adipocytes suggesting their significance in adipogenesis. Genes for multiple TRP channels were also expressed in murine WAT and BAT, out of which TRPV4, TRPV6 and TRPC6 showed differential expression. CONCLUSION Present study demonstrates the expression of TRP channels in mouse cell lines and adipose tissues.

Cinnamaldehyde supplementation prevents fasting‐induced hyperphagia, lipid accumulation, and inflammation in high‐fat diet‐fed mice

Cinnamaldehyde, a bioactive component of cinnamon, is increasingly gaining interest for its preventive and therapeutic effects against metabolic complications like type-2 diabetes. This study is an attempt to understand the effect of cinnamaldehyde in high-fat diet (HFD)-associated increase in fasting-induced hyperphagia and related hormone levels, adipose tissue lipolysis and inflammation, and selected cecal microbial count in mice. Cinnamaldehyde, at 40 µM dose, prevented lipid accumulation and altered gene expression toward lipolytic phenotype in 3T3-L1 preadipocyte cell lines. In vivo, cinnamaldehyde coadministration prevented HFD-induced body weight gain, decreased fasting-induced hyperphagia, as well as circulating leptin and leptin/ghrelin ratio. In addition to that, cinnamaldehyde altered serum biochemical parameters related to lipolysis, that is, glycerol and free fatty acid levels. At transcriptional level, cinnamaldehyde increased anorectic gene expression in hypothalamus and lipolytic gene expression in visceral white adipose tissue. Furthermore, cinnamaldehyde also decreased serum IL-1β and inflammatory gene expression in visceral white adipose tissue. However, cinnamaldehyde did not modulate the population of selected gut microbial (Lactobacillus, Bifidibaceria, and Roseburia) count in cecal content. In conclusion, cinnamaldehyde increased adipose tissue lipolysis, decreased fasting-induced hyperphagia, normalized circulating levels of leptin/ghrelin ratio, and reduced inflammation in HFD-fed mice, which augurs well for its antiobesity role.

Survival and synergistic growth of mixed cultures of bifidobacteria and lactobacilli combined with prebiotic oligosaccharides in a gastrointestinal tract simulator.

Background Probiotics, especially in combination with non-digestible oligosaccharides, may balance the gut microflora while multistrain preparations may express an improved functionality over single strain cultures. In vitro gastrointestinal models enable to test survival and growth dynamics of mixed strain probiotics in a controlled, replicable manner. Methods The robustness and compatibility of multistrain probiotics composed of bifidobacteria and lactobacilli combined with mixed prebiotics (galacto-, fructo- and xylo-oligosaccharides or galactooligosaccharides and soluble starch) were studied using a dynamic gastrointestinal tract simulator (GITS). The exposure to acid and bile of the upper gastrointestinal tract was followed by dilution with a continuous decrease of the dilution rate (de-celerostat) to simulate the descending nutrient availability of the large intestine. The bacterial numbers and metabolic products were analyzed and the growth parameters determined. Results The most acid- and bile-resistant strains were Lactobacillus plantarum F44 and L. paracasei F8. Bifidobacterium breve 46 had the highest specific growth rate and, although sensitive to bile exposure, recovered during the dilution phase in most experiments. B. breve 46, L. plantarum F44, and L. paracasei F8 were selected as the most promising strains for further studies. Conclusions De-celerostat cultivation can be applied to study the mixed bacterial cultures under defined conditions of decreasing nutrient availability to select a compatible set of strains.

Biomineralization of hydroxyapatite in silver ion-exchanged nanocrystalline ZSM-5 zeolite using simulated body fluid

Silver ion-exchanged nanocrystalline zeolite (Ag-Nano-ZSM-5) and silver ion-exchanged conventional zeolite (Ag-ZSM-5) were synthesized. Zeolites were incubated in simulated body fluid at 310K for different time periods to grow hydroxyapatite in their matrixes. Significant large amount of hydroxyapatite was grown in Ag-Nano-ZSM-5 matrix after incubation in simulated body fluid when compared to Ag-ZSM-5. The resultant material was characterized using X-ray diffraction, N2-adsorption, scanning/transmission electron microscopy, energy dispersive X-ray, and inductively coupled plasma analysis. Mechanical properties such as compressive modulus, compressive strength, and strain at failure of the parent materials were evaluated. Biocompatibility assays suggested that Ag-Nano-ZSM-5 and hydroxyapatite grown in Ag-Nano-ZSM-5 were compatible and did not impose any toxicity to RAW 264.7 cells macrophase and Caco2 cells suggesting considerable potential for biomedical applications such as bone implants.

Isomalto-oligosaccharides, a prebiotic, functionally augment green tea effects against high fat diet-induced metabolic alterations via preventing gut dysbacteriosis in mice

&NA; High fat diet (HFD)‐induced alterations in gut microbiota and resultant ‘leaky gut’ phenomenon promotes metabolic endotoxemia, ectopic fat deposition, and low‐grade systemic inflammation. Here we evaluated the effects of a combination of green tea extract (GTE) with isomalto‐oligosaccharide (IMOs) on HFD‐induced alterations in mice. Male Swiss albino mice were fed with HFD (58% fat kcal) for 12 weeks. Systemic adiposity, gut derangement parameters and V3‐V4 region based 16S rRNA metagenomic sequencing, ectopic fat deposition, liver metabolome analysis, systemic and tissue inflammation, and energy homeostasis markers along with gene expression analysis in multiple tissues were done in mice supplemented with GTE, IMOs or their combination. The combination of GTE and IMOs effectively prevented HFD‐induced adiposity and lipid accumulation in liver and muscle while normalizing fasting blood glucose, insulin, glucagon, and leptin levels. Co‐administration of GTE with IMOs effectively modulated liver metabolome associated with lipid metabolism. It also prevented leaky gut phenotype and HFD‐induced increase in circulating lipopolysaccharides and pro‐inflammatory cytokines (e.g. resistin, TNF‐&agr;, and IL‐1&bgr;) and reduction in anti‐inflammatory cytokines (e.g. adiponectin and IL‐6). Gene expression analysis across multiple tissues further supported these functional outcomes. Most importantly, this combination improved beneficial gut microbiota (Lactobacillus sp., Bifidobacteria, Akkermansia muciniphila, Roseburia spp.) abundances, restored Firmicutes/Bacteriodetes and improved Prevotella/Bacteroides proportions. In particular, a combination of these two agents has shown improved beneficial effects on multiple parameters studied. Data presented herein suggests that strategically chosen food components might be highly effective in the prevention of HFD‐induced alterations and may further be developed as functional foods. Graphical abstract Figure. No caption available. Preventive effects of co‐administration of GTE with IMOs against HFD‐induced alterations in mice. Administration of GTE with IMOs to HFD‐fed mice prevented diet induced pathologies across multiple organ system. Its supplementation prevented adipose tissue mass building and prevented systemic obesity. It also prevented HFD‐induced alteration in insulin, glucagon and leptin levels. In liver, GTE with IMOs most effectively prevented HFD‐induced lipid and glucose metabolism aberrations, inflammation (via NF‐kB pathway activation) and metabolites patterns. Systemically, it prevented HFD‐induced increase in pro‐inflammatory cytokines and reduction in anti‐inflammatory adipokines. Its supplementation also prevented HFD‐induced impairment in glucose tolerance and in turn improved insulin sensitivity. It also prevented HFD‐induced gut microbial dysbiosis along with structural damages and SCFAs production. A reduced lipopolysaccharide production further assisted towards improved insulin sensitivity and reduced systemic inflammation.

A novel multi-strain probiotic and synbiotic supplement for prevention of Clostridium difficile infection in a murine model.

The protective effect of a multi‐strain probiotic and synbiotic formulation was evaluated in C57BL/6 mice infected with Clostridium difficile (CD) NAP1/027. Antibiotic‐treated mice were divided into the following four groups: Group 1, fed with a synbiotic formulation consisting of Lactobacillus plantarum F44, L. paracasei F8, Bifidobacterium breve 46, B. lactis 8:8, galacto‐oligosaccharides, isomalto‐oligosaccharides, and resistant starch; Group 2, fed with the same four probiotic strains as Group 1; Group 3, fed with the same prebiotic supplements as Group 1 for 7 days before CD infection; and Group 4 (control group) antibiotic treated and infected with NAP1/027 strain. Feces and cecal contents were collected for microbial cell viability, quantitative PCR (qPCR), toxin analyses and histopathology. Synbiotics‐ and probiotics‐fed mice showed a significant increase in total bifidobacteria (P < 0.05). The total lactobacilli count was increased in Group 1. Tests for cecal toxins were negative in Group 2 mice, whereas one sample each from Group 1 and 3 was positive. qPCR of cecal contents showed significant reduction in NAP1/027 DNA copies in Groups 1 and 2 and significantly higher numbers of B. breve 46, L. plantarum F44, and L. paracasei F8 in Groups 1 and 2 (P < 0.05); these changes were much less pronounced in Groups 3 and 4. Our findings indicate that the newly developed synbiotic or multi‐strain probiotic formulation confers protection against NAP1/027 infection in C57BL/6 mice. This holds promise for performing human studies.

Protective effects of phyllanthin, a lignan from Phyllanthus amarus, against progression of high fat diet induced metabolic disturbances in mice

Evidence based studies have proved the efficacy of plant derived bioactives against lifestyle oriented disorders as they can be incorporated in to the diet or diet based supplements. Phyllanthin is one such lignan from Phyllanthus amarus as well as different Phyllanthus species. Phyllanthin was evaluated as a chronic intervention (12 weeks) in mice, at a daily dose of 2 and 4 mg kg−1 of body weight along with a lard based high fat diet (HFD). Phyllanthin protected against HFD induced weight gain and adiposity. Phyllanthin supplementation reduced mRNA expression of adipogenic genes and increased expression of lipolytic genes in white adipose tissue. Treatment also showed reduction in liver triglyceride accumulation. HFD induced serum lipid disturbances were found to be restored by phyllanthin. Treatment reduced serum triglycerides and free fatty acids in HFD fed mice. Phyllanthin counteracted coexisting low grade inflammation and oxidative stress in adipose tissue and liver. Along with serum proinflammatory cytokines, expression of NF-κB and F4/80 was decreased by phyllanthin. Supplementation of phyllanthin accelerated glucose clearance along with alleviation of insulin resistance in terms of HOMA-IR. Furthermore, mRNA expression of the insulin receptor and insulin receptor substrate-1 was elevated by phyllanthin in liver and adipose tissue. The present study confirmed the protective effects of phyllanthin against HFD induced metabolic changes. Daily consumption of phyllanthin in the diet as a nutraceutical can ameliorate the development of metabolic disorders.

Anticancer Potential of Ginger: Mechanistic and Pharmaceutical Aspects

BACKGROUND Multifaceted pathologies like cancers involve multiple targets. Failure of current treatment options modulating specific tumor target, evokes need for alternate approach of either combining several smart drugs or design a dirty drug that may simultaneously influence multiple targets to trigger a cascade of protective events complementing one another. METHODS Present review tends to unravel the mechanism of anticancer action of ginger and also address issues, which may limit its realization as a biotherapeutic. RESULTS Ginger exhibits a pleiotropy of antioxidant, anti-inflammatory, antiemetic, anticancer, and antimutagenic effects. In vivo and in vitro studies have established that phenolic components of ginger, particularly 6-gingerol and 6-shogaol induce apoptosis and autophagy and inhibit metastasis. The poor biological profile of ginger extract or its actives is due to its restricted biopharmaceutical properties. The gap in manifesting the curative/therapeutic effects of these agents can be plugged by assigning them with a suitable pharmaceutical couture. CONCLUSION Hence, amalgamating the rational formulation design with observational folklore data available on herbal drugs/agents, complemented with scientific and precise in vitro and in vivo findings can bring out a class of safe, cheap, and effective curatives which can address multitarget diseases like cancers.