Protective effect of (+)-catechin against lipopolysaccharide-induced inflammatory response in RAW 264.7 cells through downregulation of NF-κB and p38 MAPK.

Catechin, a flavonol belonging to the flavonoid group of polyphenols is present in many plant foods. The present study was done to evaluate the effect of catechin on various inflammatory mediators using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The effect of catechin on total cyclooxygenase (COX) activity, 5-lipoxygenase (5-LOX), myeloperoxidase, nitrite and inducible nitric oxide synthase (iNOS) level, secretion of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were assessed in LPS-stimulated RAW 264.7 cells. The expression of COX-2, iNOS, TNF-α, nuclear factor-ĸB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) genes were also investigated. The effect was further analyzed using human PBMCs by assessing the level of TNF-α and IL-10. The study demonstrated that the inflammatory mediators such as COX, 5-LOX, nitrite, iNOS, and TNF-α were significantly inhibited by catechin in a concentration-dependent manner whereas IL-10 production was up-regulated in RAW 264.7 cells. Moreover, catechin down-regulated the mRNA level expression of COX-2, iNOS, TNF-α, NF-κB and p38 MAPK. The current study ratifies the beneficial effect of catechin as a dietary component in plant foods to provide protection against inflammatory diseases.

Bacterial endophytes from Artemisia nilagirica (Clarke) Pamp., with antibacterial efficacy against human pathogens.

A study was conducted to isolate and characterize endophytes from Artemisia nilagirica, a traditional medicinal plant. The plant was collected from Western Ghats, India. Endophytes isolated included Arthrobacter sp. WWAT1, Pseudomonas sp. WYAT2, Microbacterium sp. WYAT3, Psychrobacter sp. WBAT4, Enterobacter sp. WWAT5, Bacillus sp. WBAT6, Kosakonia cowanii WBAT7, Bacillus sp. WBAT8, Bacillus sp. WBAT9, Chromobacterium violaceum WVAT6, Serratia sp.WPAT8 and Burkholderia sp. WYAT7. Of these two bacteria, Chromobacterium violaceum strain WVAT6 and Burkholderia sp. strain WYAT exhibited antibacterial property against human pathogens. Similar to the environmental isolates, Burkholderia sp. WYAT7 showed pleomorphism and produced different enzymes, whereas like clinical strains they showed multidrug resistance, for their survival in different environmental conditions. Chromobacterium violaceum WVAT6 exhibited rod shape morphology and showed multiple drug resistance except to erythromycin, tetracycline and gentamicin antibiotics. Both produced biofilm and enzymes such as protease and lipase. The antimicrobial compounds from these endophytes may find application in the preparation of antimicrobial formulations.

Catechin rich butanol fraction extracted from Acacia catechu L. (a thirst quencher) exhibits immunostimulatory potential.

Acacia catechu L., (Fabaceae) named as "catechu" is a plant, the decoction of heartwood of which is daily consumed as thirst quencher by a good percentage of the population in South India. The plant is mainly distributed in India and other Asian countries. It has been used in Indian traditional medicine for the treatment of asthma, bronchitis, colic, diarrhea, boils, skin afflictions, sores and stomatitis. The present investigation was aimed to study the immunomodulatory effects of different fractions of ethanol extract of A. catechu heartwood and HPLC analysis of the active fraction. Three fractions namely, butanol, chloroform and ethyl acetate were prepared from ethanol extract of A. catechu heartwood. Each of these fractions was assessed for its immunomodulatory activity. In vivo immunomodulatory activity was analyzed by sheep red blood cells (SRBC) specific hemagglutinating antibody titer, plaque-forming cell assay and delayed type hypersensitivity (DTH) reaction in Swiss albino mice. In vitro immunomodulating potential of the fractions was studied using murine peritoneal macrophages and splenocytes. Non-specific immune functions such as phagocytosis (nitroblue tetrazolium reduction assay and cellular lysosomal enzyme assay), nitric oxide (NO) production and cytokine release (TNF-α and IL-10) were studied in macrophages. In addition, splenocyte proliferation was also studied. In the in vivo experiments, butanol and chloroform fractions showed an increase in antibody titer dose-dependently. At higher dose (400 mg/kg b. w.) treatment the butanol fraction produced an enhancement in the number of plaque-forming cells (antibody producing cells) in the spleen. SRBC induced DTH reaction was significantly increased with butanol fraction in a dose-dependent manner. Peritoneal macrophages showed an increased phagocytic response on treatment with butanol fraction (100 µg/mL) as evidenced by its effect on nitroblue tetrazolium reduction and cellular lysosomal enzyme activity. All three fractions inhibited the production of NO and the release of TNF-α. Interleukin-10 production was significantly increased after treatment with butanol fraction. High-performance liquid chromatography analysis of the butanol fraction showed the presence of high concentration of catechin. The results suggested that butanol fraction of ethanol extract of A. catechu heartwood had immunomodulatory effects on non-specific, humoral, and cell-mediated immune functions. This study may be useful in validating the rationality of daily consumption of decoction of A. catechu and also its use in traditional medicine system. The study also suggests the possible use of A. catechu in the immunostimulatory herbal preparations.

Immunomodulatory activities of Acacia catechu, a traditional thirst quencher of South India.

BACKGROUND: Acacia catechu has been widely used in Ayurveda for treating many diseases. Its heartwood extract is used in asthma, cough, bronchitis, colic, diarrhea, dysentery, boils, skin afflictions, sores and for stomatitis. The decoction of heartwood is used for drinking purpose in southern part of India especially in Kerala. OBJECTIVE: The current study was carried out to evaluate immunomodulatory effects of heartwood extracts of A. catechu in Swiss albino mice. MATERIAL AND METHODS: In vivo immunomodulatory activity was analyzed by hemagglutinating antibody (HA) titer, plaque forming cell assay and delayed type hypersensitivity (DTH). In vitro immunomodulatory potential of the extracts was studied using peritoneal macrophages and splenocytes from mice. Effect of extracts on phagocytic activity of macrophages was analyzed by nitroblue tetrazolium (NBT) reduction assay and cellular lysosomal enzyme assay. Anti-inflammatory activity was studied by nitric oxide (NO) assay and production of TNF-α and IL-10. RESULTS: A dose dependent increase in antibody titer was observed with extracts treatment. Treatment with extracts produced an enhancement in the number of antibody producing cells in the spleen. DTH reaction was significantly decreased with extracts treatment. An increased phagocytic response was shown by peritoneal macrophages on treatment with the extracts as evidenced by its effect on NBT reduction and cellular lysosomal enzyme activity. The extracts inhibited the release of pro-inflammatory cytokine TNF-α and the production of NO. IL-10 production was significantly increased after extract treatment. CONCLUSION: The results of the present study indicate the immunomodulatory effects of A. catechu extracts on humoral, cell mediated and non-specific immune functions.

Efficient visible light induced synthesis of silver nanoparticles by Penicillium polonicum ARA 10 isolated from Chetomorpha antennina and its antibacterial efficacy against Salmonella enterica serovar Typhimurium.

The green synthesis of silver nanoparticles (AgNPs) using biological systems such as fungi has evolved to become an important area of nanobiotechnology. Herein, we report for the first time the light-induced extracellular synthesis of silver nanoparticles using algicolous endophytic fungus Penicillium polonicum ARA 10, isolated from the marine green alga Chetomorpha antennina. Parametric optimization, including the concentration of AgNO3, fungal biomass, ratio of cell filtrate and AgNO3, pH, reaction time and presence of light, was done for rapid AgNPs production. The obtained silver nanoparticles (AgNPs) were characterized by UV-Visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy and Transmission electron microscopy (HRTEM-EDAX). The AgNPs showed a characteristic UV-visible peak at 430 nm with an average size of 10-15 nm. The NH stretches in FTIR indicate the presence of protein molecules. The Raman vibrational bands suggest that the molecules responsible for the reduction and stability of AgNPs were extracellular proteins produced by P.polonicum. Antibacterial evaluation of AgNPs against the major foodborne bacterial pathogen Salmonella enterica serovar Typhimurium MTCC 1251, was assessed by well diffusion, Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assay. Killing kinetic studies revealed complete killing of the bacterial cells within 4 h and the bactericidal nature of synthesized nanoparticles was confirmed by fluorescent microscopy and scanning electron microscopy. Furthermore, the bactericidal studies with Transmission electron microscopy (TEM) at different time intervals explored the presence of AgNPs in the cell wall of S.Typhimurium at about 30 min and the complete bacterial lysis was found at 24 h. The current research opens an insight into the green synthesis of AgNPs and the mechanism of bacterial lysis by direct damage to the cell wall.

Antibacterial activity of autochthonous bacteria isolated from Anabas testudineus (Bloch, 1792) and it's in vitro probiotic characterization.

Antimicrobial potentials of bacteria isolated from Anabas testudineus have been evaluated through in vitro antagonistic activity against potent fish pathogens. The cellular components and filtered culture medium were effective against six fish pathogens. Altogether 110 strains were isolated from the fish gut, out of which 10 strains were selected through well diffusion method. From them, a strain HGA8B having cumulative maximum score was selected as candidate probiotic. The whole-cell product, heat-killed whole-cell product, Ethyl acetate extract, and the filtered broth were exhibited bactericidal activity against the tested pathogens. In addition, the isolated bacterium was capable of producing extracellular enzymes important for the digestion of food materials and was capable of growth in fish mucus from Oreochromis niloticus. The strain tolerated bile juice secreted by the host and effectively produced biofilm. Analysis of 16S rDNA sequence revealed that isolated strain HGA8B was Bacillus sp. (MF351637). Furthermore, intraperitoneal injection of the bacterium did not induce any pathological signs, symptoms or mortalities in Oreochromis niloticus and revealed the safety of this bacterium as a candidate probiotic in aquaculture.