Srisesharam Srigopalram

In vitro importance of probiotic Lactobacillus plantarum related to medical field.

Lactobacillus plantarum is a Gram positive lactic acid bacterium commonly found in fermented food and in the gastro intestinal tract and is commonly used in the food industry as a potential starter probiotic. Recently, the consumption of food together with probiotics has tremendously increased. Among the lactic acid bacteria, L. plantarum attracted many researchers because of its wide applications in the medical field with antioxidant, anticancer, anti-inflammatory, antiproliferative, anti-obesity and antidiabetic properties. The present study aimed to investigate the in vitro importance of L. plantarum toward medical applications. Moreover, this report short listed various reports related to the applications of this promising strain. In conclusion, this study would attract the researchers in commercializing this strain toward the welfare of humans related to medical needs.

Potential Application of p-Coumaric Acid on Differentiation of C2C12 Skeletal Muscle and 3T3-L1 Preadipocytes-An in Vitro and in Silico Approach.

Coumaric acid (CA) is a phenolic acid of the hydroxycinnamic acid family, and it has many biological functions such as anti-oxidant, anti-inflammatory, antidiabetic, anti-ulcer, anti-platelet, anti-cancer activities, etc. In the present study, we planned to analyse the potential molecular function of CA on skeletal muscle and preadipocytes differentiation using PCR and Western blot techniques. First, we analysed the impact of CA on C2C12 skeletal muscle differentiation. It revealed that CA treatment inhibited horse serum-induced skeletal muscle differentiation as evidenced by the decreased expression of early myogenic differentiation markers such as Myogenin and myoD via the AMP activated protein kinase- alpha AMPK-α mediated pathway. Furthermore, the level of lipid accumulation and changes in genes and protein expressions that are associated with lipogenesis and lipolysis were analyzed in 3T3-L1 cells. The Oil Red O staining evidenced that CA treatment inhibited lipid accumulation at the concentration of 0.1 and 0.2 mM. Furthermore, coumaric acid treatment decreased the expression of main transcriptional factors such as CCAAT/enhancer binding protein-alpha (C/EBP-α) and peroxisome proliferator-activated receptor gamma-2 (PPAR-γ2). Subsequently, CA treatment decreased the expression of sterol regulatory element binding protein-1 (SREBP-1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC) and adiponectin. Finally, we identified conformational changes induced by CA in PPAR-γ2 using computational biology tools. It revealed that CA might downregulate the PPAR-γ2 expression by directly binding with amino acids of PPAR-γ2 by hydrogen at 3.26 distance and hydrophobic interactions at 3.90 contact distances. These data indicated that CA suppressed skeletal muscle and preadipocytes differentiation through downregulation of the main transcriptional factors and their downstream targets.

Isolation, In Vitro Probiotic Characterization of Lactobacillus plantarum and its Role on Italian ryegrass Silage Quality Enhancement

Italian ryegrass (IRG) is an important forage type for feeding ruminants. IRG has difficult to ensile with good quality. Hence the inoculation IRG with lactic acid producing bacterial strains (LAB) gives an alternate solution to produce quality silage. Accordingly new Lactobacillus sp. KCC-32 was isolated from fermented animal manure. Biochemical and physiological studies showed that the strain belonged to Gram positive, produced gas from glucose and catalase-negative. The 16S rRNA sequence analysis revealed that KCC-32 showed 99% similarity towards Lactobacillus plantarum sp. Further, KCC-32 displayed potential probiotic characteristics including resistant to low pH, bile salt tolerance, auto-aggregation and hydrophobicity. The homo fermentative activity of KCC-32 resulted in the enhancement of Italian ryegrass silage quality. In addition, KCC-32 added silage group showed significantly (P≤0.05) increased lactic acid production (4.891 DM%) and the nutrient profile resulted with high crude protein content with less acid detergent fiber (ADF) and neutral detergent fiber (NDF) percentage when compared to control group. Further, the microbial count of KCC-32 silage group displayed significantly (P≤0.05) high LAB count (24.3×10 7 cfu/g) and no fungal as well as yeast growth. Hence, this study suggests that KCC-32 has potential probiotic characteristics and the addition of KCC-32 to the IRG silage can improve the fermentation quality for the production of high-quality silage.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach

BackgroundFrom ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipogenic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes.ResultsWe analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC–MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-γ2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ2, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells.ConclusionThe present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.

Proteomic Analysis of Stage-II Breast Cancer from Formalin-Fixed Paraffin-Embedded Tissues

Breast cancer is the most frequently occurring disease among women worldwide. The early stage of breast cancer identification is the key challenge in cancer control and prevention procedures. Although gene expression profiling helps to understand the molecular mechanism of diseases or disorder in the living system, gene expression pattern alone is not sufficient to predict the exact mechanisms. Current proteomics tools hold great application for analysis of cancerous conditions. Hence, the generation of differential protein expression profiles has been optimized for breast cancer and normal tissue samples in our organization. Normal and tumor tissues were collected from 20 people from a local hospital. Proteins from the diseased and normal tissues have been investigated by 2D gel electrophoresis and MALDI-TOF-MS. The peptide mass fingerprint data were fed into various public domains like Mascot, MS-Fit, and Pept-ident against Swiss-Prot protein database and the proteins of interest were identified. Some of the differentially expressed proteins identified were human annexin, glutathione S-transferase, vimentin, enolase-1, dihydrolipoamide dehydrogenase, glutamate dehydrogenase, Cyclin A1, hormone sensitive lipase, beta catenin, and so forth. Many types of proteins were identified as fundamental steps for developing molecular markers for diagnosis of human breast cancer as well as making a new proteomic database for future research.

Beneficial Effects of Lactic Acid Bacteria Inoculation on Oat Based Silage in South Korea

The objective of the study was to measure the beneficial effects of lactic acid bacteria (LAB) inoculation on the nutritive value of oat silage collected from thirteen regions in the Republic of Korea. The contents of crude protein, acid detergent fiber (ADF), neutral detergent fiber (NDF) and crude ash (CA) were slightly lower in LAB inoculated silage when compared with the control silage, whereas inoculation of LAB resulted in increased total digestible nutrient (TDN). Higher number of LAB, but lower count of yeast and fungi indicated the effectiveness of the LAB inoculation on oat silage fermentation. LAB inoculation resulted in low pH silage, which may prevent undesirable microbial growth. The LAB inoculation promoted lactic acid dominant fermentation with marginal levels of acetic acid and butyric acid in oat silage. These data suggest that the LAB inoculation may preserve oat silage at better quality for ruminant animal production.

Addition of Novel Lactobacillus plantarum KCC-10 and KCC-19 to Improve Fermentation Quality and Characterization of Italian Ryegrass Silage

ABSTRACT An investigation was carried out to evaluate the potential role of Lactobacillus Plantarum KCC-10 and KCC-19 on the quality and fermentation characterization of Italian ryegrass (IRG) silages. The physiochemical properties of IRG silage such as crude protein content, neutral detergent fiber, acid detergent fiber, total digestible nutrient and in vitro dry matter digestibility were not affected by KCC-10 and KCC-19. The pH of IRG silage in KCC-10 and KCC-19 treatments decreased compared to the control (p<0.05), while the lactic acid content in KCC-10 and KCC-19 treatments increased compared to the control (p<0.05). In addition, the number of lactic acid bacteria (LAB) in the KCC-10 treatment increased compared to the control (p<0.05). The number of lactic acid bacteria in KCC-19 increased, but there was no significant difference in all treatments. Therefore, we recommend L. plantarum KCC-10 and KCC-19 as potential additive candidates in IRG silage with lots of advantages.(

Silages of Rye Harvested at Different Stages: A Study on Microbial Inoculants Responses in Improving Rye Silage Fermentation Quality

The present study analyzes the role of Lactic Acid Bacteria Mixture (LBM) on improving rye silage quality. Rye of four different stages (Booting, Heading, Flowering, and Late flowering) was collected and silage was prepared. The nutrient profile analysis of experimental silage groups showed no significant changes between control and LBM inoculation. Interestingly, the pH of rye silage in LBM treatments showed significant reduction than control (p<0.05) in all stages of rye silage. However, lowest pH (3.69) resulted on booting stage among other stages of rye. Subsequently significant lactic acid production was noted in all stages of LBM inoculation than control. Conversely maximum lactic acid production of (5.33%DM) was noted at booting stage followed by (4.86%DM) in heading stage. Further the lactic acid bacterial (LAB) count in LBM inoculated group showed significant increase than control. Similarly, the silage of booting stage group registered maximum LAB population (63.7 ×106CFU/g) after that heading stage (32.3 ×106CFU/g). Further significant reduction in yeast growth and no fungal growth was noted in all LPM treatment groups. Hence, LBM inoculants could be a better additive for improving rye silage quality.

Effect of Lactic Acid Bacteria Inoculation on Fermentation Characteristics of Whole Crop Barley Silage

An experiment was carried out to determine the homofermentative activity of Lactobacillus plantarum KCC-10 and KCC-19 on the ensiling of whole crop barley (WCB). The crude protein in the silages was slightly higher in the KCC-10 and KCC-19 treatments compared to the control, but there was no significant difference between the two inoculant-treated silages. Nutrient parameters such as acid detergent fiber, neutral detergent fiber and in vitro dry matter digestibility in L. plantarum KCC-10 and KCC-19 treated silages did not differ from those in the control silage. The lactic acid content increased in KCC-10 and KCC-19 treated silage when compared with the control silage but the contents of acetic acid and butyric acid produced in KCC-10 and KCC-19 treated silages were similar with the control silage. Further, the number of lactic acid bacteria (LAB) in KCC-10 treated silage demonstrated a significant increase when compared to the control. Especially, KCC-19 treated silage showed greater lactic acid bacterial growth potential. Other microbes such as yeast and fungi were not detected in KCC-10 and KCC-19 treated WCB silages. Hence, this study suggests that the addition of L. Plantarum KCC-10 and KCC-19 to the WCB silage can improve fermentation quality for the production of high-quality silage.