Correlation of attention deficit hyperactivity disorder with gut microbiota according to the dietary intake of Korean elementary school students.

We investigated the impact of dietary patterns on the gut microbiota and concentration of short-chain fatty acids in the feces of Korean elementary school students. The dietary intake and ADHD assessment of 40 Korean elementary school students were analyzed using a dish-based semi-quantitative food frequency questionnaire. Analysis of gut microbiota and short-chain fatty acids composition were performed using the real-time polymerase chain reaction, metagenomics, and gas chromatography methods. The dietary patterns of participants were divided into four groups: healthy, processed food, fish and shellfish, and meat. The participants were also divided into two groups according to their ADHD scores: 0-30, control group; over 30, ADHD group. The ADHD score of the processed food group was significantly higher than that of the healthy group. The processed food and ADHD groups showed significantly higher abundance of harmful bacteria, such as the Enterobacter, Escherichia coli, and Clostridium strains, and markedly lower abundance of beneficial bacteria, such as the Bifidobacterium and Ruminococcus strains, than the control group. The heat maps of metagenomics indicated that each group was separated into distinct clusters, and the processed food and ADHD groups showed significantly lower α-diversity of gut microbiota than the control group. In these groups, the concentration of acetate or butyrate in the feces was significantly lower than that in the control group. These results may indicate that imbalanced diets can disturb the colonic microbial balance and are likely to become a potential risk factor for the prevalence of ADHD.

Butyrate modulates mucin secretion and bacterial adherence in LoVo cells via MAPK signaling.

Short-chain fatty acids contribute to normal bowel function and prevent bacterial infections. In particular, butyrate is a promising candidate that plays an important role in regulating the functional integrity of the gastrointestinal tract by stimulating mucin secretion. We investigated whether butyrate treatment modulates mucin secretion and bacterial adherence in LoVo cells. In addition, the possible signaling pathways were also examined in connection with the upregulation of mucin secretion. The results showed that butyrate induced mucin secretion in LoVo cells, resulting in the inhibition of Escherichia coli adhesion by increasing the adherence of Lactobacillus acidophilus and Bifidobacterium longum. The gene expression analysis suggests that mitogen-activated protein kinase (MAPK) signaling pathways including Cdc42-PAK pathway appears to be involved in stimulating mucin secretion. More importantly, butyrate induced the increased actin expression and polymerization in LoVo cells, which could be attributable to the Cdc42-PAK signaling pathway, implicated in actin cytoskeleton and mucin secretion. Our results provide a molecular basis in modulating bacterial adherence and the MAPK signaling pathway for the improved homeostasis of colonic epithelial cells.

Quality Characteristics of Substitute Meat Patties Developed Using Aruncus dioicus var. kamtschaticus Hara.

We developed a vegetable alternative to meat patties using Aruncus dioicus var. kamtschaticus Hara (A. dioicus) and used it to generate basic data for the alternative meat market by comparing nutritional and microbiological components with commercially available vegetable and meat patties. Nutrient analysis, microbiological analysis, chromaticity, and texture analysis were performed on substitute meat patties (SMPs) with A. dioicus and commercially available vegetable and animal patties. Among sugars, the contents of fructose and maltose were respectively high in commercial meat patties (CMPs) and SMPs. SMPs were low in saturated and trans-fat, and high in ω-3 fatty acids. The contents (in descending order) of leucine > phenylalanine > threonine > isoleucine were high in SMPs and commercial vegetable patties (CVPs). Qualitative and quantitative findings of Escherichia coli (E. coli) and Staphylococcus aureus were all negative. Our SMPs had high lightness (L*), low redness (a*), and low yellowness (b*). The hardness, chewiness, and resilience of our SMPs were lower than those of other vegetable and animal patties. Considering our results, the method of manufacturing SMPs developed in the present study allows meat to be flavored without significant nutritional differences compared with commercially available CMPs. Our findings provide a base for studies on future meat alternatives.

Imbalanced dietary intake alters the colonic microbial profile in growing rats.

An imbalanced dietary intake is associated with alteration of intestinal ecosystem. We investigated the impact of imbalanced diets on colonic microbiota, concentrations of short chain fatty acid in colonic digesta and serum immunoglobulins (Igs) of growing rats. Compared to the control diet, consuming diets high in fat, sucrose, or processed meat, or low in iron, increased the abundance of the pathogenic bacteria such as Clostridium, Escherichia coli, and Salmonella species, and decreased the beneficial bacteria, like Bifidobacteria, Lactobacillus, Akkermansia, Phascolarctobacterium, Alistipes, and butyrate producing species of bacteria in the colon of growing rats. The heatmap of metagenomics indicated that each group was separated into distinct clusters, and the ID group in particular, showed significantly (P < 0.01) reduced alpha diversity of colonic microbiota in comparison to the control group. All experimental groups showed significantly (P < 0.05 or P < 0.01) decreased concentration of acetate and butyrate in the colonic digesta and lower levels of serum IgG or IgA, compared to the control. These results indicated that the imbalanced dietary intake negatively altered intestinal ecosystem and immunity.

Development of livestock feed additives using porcine blood cells.

Livestock blood discarded during slaughtering has potentially valuable components such as plasma proteins and haemoglobin. Plasma is used as a feed additive following processing via different methods, including spray drying, whereas blood cells have been underutilized. In this study, we developed haemoglobin hydrolysate (HH) and iron-enriched residue (IER) from porcine blood cells and investigated whether their oral administration regulates the immune system and gut microbiota in growing rats. Twenty-one Sprague-Dawley male rats (n = 7) were used during a 4-week trial and were fed a control, HH or IER diet. The ratio of beneficial bacteria such as Lactobacillus and Akkermansia strains increased in rats fed HH or IER diets. Moreover, compared with the control group, the IER group had an elevated ratio of Lactobacillus to Enterobacteria, which is regarded as an index of beneficial aspect in the gut. Phagocytosis of peripheral blood leucocytes was higher in the HH and IER groups than in the control group. The level of plasma immunoglobulin G increased to approximately 72.7 mg/ml and 152.0 mg/ml in the HH and IER groups, respectively, which was significantly (p < 0.05) higher than that in the control group. These results confirm that HH and IER developed in this study may be a potential additive for animal feeds.

Manufacture and Physicochemical Properties of Chitosan Oligosaccharide/A2 ß-Casein Nano-Delivery System Entrapped with Resveratrol.

The purposes of this research were to form chitosan oligosaccharide (CSO)/A2 ß-casein nano-delivery systems (NDSs) and to investigate the effects of production variables, such as CSO concentration levels (0.1%, 0.2%, and 0.3%, w/v) and manufacturing temperature (5°C, 20°C, and 35°C), on the production and physicochemical characteristics of CSO/A2 ß-casein NDSs to carry resveratrol. The morphological characteristics of CSO/A2 ß-casein NDSs were assessed by the use of transmission electron microscopy (TEM) and particle size analyzer. High-performance liquid chromatography (HPLC) was applied to determine the entrapment efficiency (EE) of resveratrol. In the TEM images, globular-shaped particles with a diameter from 126 to 266 nm were examined implying that NDSs was successfully formed. As CSO concentration level was increased, the size and zeta-potential values of NDSs were significantly (p<0.05) increased. An increase in manufacturing temperature from 5°C to 35°C resulted in a significant (p<0.05) increase in the size and polydispersity index of NDSs. Over 85% of resveratrol was favorably entrapped in CSO/A2 ß-casein NDSs. The entrapment efficiency (EE) of resveratrol was significantly (p<0.05) enhanced with an increase in manufacturing temperature while CSO concentration level did not significantly affect EE of resveratrol. There were no significant (p<0.05) changes observed in the size and polydispersity index of NDSs during heat treatments and storage in model milk and yogurt indicating that CSO/A2 ß-casein NDSs exhibited excellent physical stability. In conclusion, the CSO concentration level and manufacturing temperature were the crucial determinants affecting the physicochemical characteristics of CSO/A2 ß-casein NDSs containing resveratrol.

Enzymatic Hydrolysis of Egg White Protein Exerts a Hypotensive Effect in Spontaneously Hypertensive Rats.

This study was conducted to investigate the hypotensive effect of egg white protein (EWP) hydrolysate (EWH) in spontaneously hypertensive rats (SHRs). The hydrolysis of EWP was effectively performed with a combination of 0.5% bromelain and 1% papain at 50°C for 60 min. The resulting hydrolysate did not elicit an allergic reaction as confirmed by human mast cell activation test. The systolic and diastolic blood pressures of the SHRs fed the EWH diet were observed to be significantly or numerically lower than those of the other groups during the experimental period of 28 d. EWH treatment significantly (p<0.05) upregulated the nitric oxide levels in hCMEC/D3 cells and the plasma of the SHRs compared to those in the control. Moreover, EWH ingestion significantly (p<0.01) reduced the plasma angiotensin II level of the SHRs compared with that in the control. In conclusion, beyond its basic nutritional value, EWH prevents and manages hypertension, and thus can be an invaluable resource for functional food development.

Evaluation of the Digestibility of Korean Hanwoo Beef Cuts Using the in vitro Physicochemical Upper Gastrointestinal System.

The aim of this study was to investigate the digestibility of different Korean Hanwoo beef cuts using an in vitro digestion model, in vitro physicochemical upper gastrointestinal system (IPUGS). The four most commonly consumed cuts - tenderloin, sirloin, brisket and flank, and bottom round - were chosen for this study. Beef samples (75 g) were cooked and ingested into IPUGS, which was composed of mouth, esophagus, and stomach, thereby simulating the digestion conditions of humans. Digested samples were collected every 15 min for 4 h of simulation and their pH monitored. Samples were visualized under a scanning electron microscope (SEM) to examine changes in the smoothness of the surface after digestion. Analysis of the amino acid composition and molecular weight (MW) of peptides was performed using reverse-phase high performance liquid chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively. Following proteolysis by the gastric pepsin, beef proteins were digested into peptides. The amount of peptides with higher MW decreased over the course of digestion. SEM results revealed that the surface of the digested samples became visibly smoother. Total indispensable and dispensable amino acids were the highest for the bottom round cut prior to digestion simulation. However, the total amount of indispensable amino acids were maximum for the tenderloin cut after digestion. These results may provide guidelines for the elderly population to choose easily digestible meat cuts and products to improve their nutritional and health status.

Purification and Anti-pathogenic Properties of Immunoglobulin Concentrates from Porcine Blood.

During slaughtering, animal blood is typically discarded, resulting in water pollution. However, this discarded blood has valuable components, such as immunoglobulin (Ig). Although several studies have been conducted to develop methods for effective recycling of slaughterhouse blood, they have not been commercially utilized in Korea. Here, we extracted an Ig-rich fraction from porcine blood that was then subjected to various in vitro tests, including pathogen growth inhibition, antigenic cross-reactivity, and anti-toxin activity. The porcine immunoglobulin concentrate (PIC) was effectively purified by eliminating other components, such as albumin, and consisted of approximately 63.2±2.9% IgG and 7.2±0.4% IgM on a protein basis. The results showed that it significantly suppressed the growth of pathogenic bacteria, and bound to all tested pathogens, including both gram-positive and gram-negative species, although the degree of activity differed according to strain. The PIC bound to two types of lipopolysaccharide (LPS) obtained from Escherichia coli O111:B4 and Salmonella enterica serotype typhimurium in a concentration-dependent manner. In addition, the PIC restored the proliferation activity of the lymphoblast K-562 cells when co-incubated with pathogenic LPS. These results confirm that the PIC prepared in this study is a potentially valuable functional food material or diet supplement as an alternative to antibiotics that can protect animals from pathogenic bacteria.

Hypoallergenic and Physicochemical Properties of the A2 ß-Casein Fractionof Goat Milk.

Goat milk has a protein composition similar to that of breast milk and contains abundant nutrients, but its use in functional foods is rather limited in comparison to milk from other sources. The aim of this study was to prepare a goat A2 ß-casein fraction with improved digestibility and hypoallergenic properties. We investigated the optimal conditions for the separation of A2 ß-casein fraction from goat milk by pH adjustment to pH 4.4 and treating the casein suspension with calcium chloride (0.05 M for 1 h at 25°C). Selective reduction of ß-lactoglobulin and αs-casein was confirmed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography. The hypoallergenic property of A2 ß-casein fraction was examined by measuring the release of histamine and tumor necrosis factor alpha from HMC-1 human mast cells exposed to different proteins, including A2 ß-casein fraction. There was no significant difference in levels of both indicators between A2 ß-casein treatment and the control (no protein treatment). The A2 ß-casein fraction is abundant in essential amino acids, especially, branched-chain amino acids (leucine, valine, and isoleucine). The physicochemical properties of A2 ß-casein fraction, including protein solubility and viscosity, are similar to those of bovine whole casein which is widely used as a protein source in various foods. Therefore, the goat A2 ß-casein fraction may be useful as a food material with good digestibility and hypoallergenic properties for infants, the elderly, and people with metabolic disorders.

Hydrolysis by Alcalase Improves Hypoallergenic Properties of Goat Milk Protein.

Goat milk is highly nutritious and is consumed in many countries, but the development of functional foods from goat milk has been slow compared to that for other types of milk. The aim of this study was to develop a goat milk protein hydrolysate (GMPH) with enhanced digestibility and better hypoallergenic properties in comparison with other protein sources such as ovalbumin and soy protein. Goat milk protein was digested with four commercial food-grade proteases (separately) under various conditions to achieve the best hydrolysis of αs -casein and ß-lactoglobulin. It was shown that treatment with alcalase (0.4%, 60℃ for 30 min) effectively degraded these two proteins, as determined by SDS-PAGE, measurement of nonprotein nitrogen content, and reverse-phase high-performance liquid chromatography. Hydrolysis with alcalase resulted in a significant decrease in ß-lactoglobulin concentration (almost to nil) and a ~40% reduction in the level of αs-casein. Quantification of histamine and TNF-α released from HMC-1 cells (human mast cell line) showed that the GMPH did not induce an allergic response when compared to the control. Hence, the GMPH may be useful for development of novel foods for infants, the elderly, and convalescent patients, to replace cow milk.

Physicochemical Characterization and Potential Prebiotic Effect of Whey Protein Isolate/Inulin Nano Complex.

The purposes of this study were to investigate the impacts of concentration levels of whey protein isolate (WPI) and inulin on the formation and physicochemical properties of WPI/inulin nano complexes and to evaluate their potential prebiotic effects. WPI/inulin nano complexes were produced using the internal gelation method. Transmission electron microscopy (TEM) and particle size analyzer were used to assess the morphological and physicochemical characterizations of nano complexes, respectively. The encapsulation efficiency of resveratrol in nano complexes was studied using HPLC while the potential prebiotic effects were investigated by measuring the viability of probiotics. In TEM micrographs, the globular forms of nano complexes in the range of 10 and 100 nm were successfully manufactured. An increase in WPI concentration level from 1 to 3% (w/v) resulted in a significant (p<0.05) decrease in the size of nano complexs while inulin concentration level did not affect the size of nano complexes. The polydispersity index of nano complexes was below 0.3 in all cases while the zeta-potential values in the range of -2 and -12 mV were observed. The encapsulation efficiency of resveratrol was significantly (p<0.05) increased as WPI and inulin concentration levels were increased from 1 to 3% (w/v). During incubation at 37℃ for 24 h, WPI/inulin nano complexes exhibited similar viability of probiotics with free inulin and had significantly (p<0.05) higher viability than negative control. In conclusions, WPI and inulin concentration levels were key factors affecting the physicochemical properties of WPI/inulin nano complexes and had potential prebiotic effect.

Butyrate modulates bacterial adherence on LS174T human colorectal cells by stimulating mucin secretion and MAPK signaling pathway.

BACKGROUND/OBJECTIVES: Fermentation of dietary fiber results in production of various short chain fatty acids in the colon. In particular, butyrate is reported to regulate the physical and functional integrity of the normal colonic mucosa by altering mucin gene expression or the number of goblet cells. The objective of this study was to investigate whether butyrate modulates mucin secretion in LS174T human colorectal cells, thereby influencing the adhesion of probiotics such as Lactobacillus and Bifidobacterium strains and subsequently inhibiting pathogenic bacteria such as E. coli. In addition, possible signaling pathways involved in mucin gene regulation induced by butyrate treatment were also investigated. MATERIALS/METHODS: Mucin protein content assay and periodic acid-Schiff (PAS) staining were performed in LS174T cells treated with butyrate at various concentrations. Effects of butyrate on the ability of probiotics to adhere to LS174T cells and their competition with E. coli strains were examined. Real time polymerase chain reaction for mucin gene expression and Taqman array 96-well fast plate-based pathway analysis were performed on butyrate-treated LS174T cells. RESULTS: Treatment with butyrate resulted in a dose-dependent increase in mucin protein contents in LS174T cells with peak effects at 6 or 9 mM, which was further confirmed by PAS staining. Increase in mucin protein contents resulted in elevated adherence of probiotics, which subsequently reduced the adherent ability of E. coli. Treatment with butyrate also increased transcriptional levels of MUC3, MUC4, and MUC12, which was accompanied by higher gene expressions of signaling kinases and transcription factors involved in mitogen-activated protein kinase (MAPK) signaling pathways. CONCLUSIONS: Based on our results, butyrate is an effective regulator of modulation of mucin protein production at the transcriptional and translational levels, resulting in changes in the adherence of gut microflora. Butyrate potentially stimulates the MAPK signaling pathway in intestinal cells, which is positively correlated with gut defense.

In Vitro Effects of Dietary Inulin on Human Fecal Microbiota and Butyrate Production.

Administration of dietary fibers has various health benefits, mainly by increasing numbers of beneficial bacteria and enhancing production of short-chain fatty acids in the colon. There has been growing interest in the addition of dietary fiber to human diet, due to its prebiotic effects. This study aimed to evaluate the prebiotic activity of inulin using an in vitro batch fermentation system with human fecal microbiota. Fermentation of inulin resulted in a significantly greater ratio of Lactobacillus or Bifidobacteria to Enterobacteria strains as an index of healthy human intestine and elevated butyrate concentration, which are related to improvement of gut health.

Korean ginseng modulates the ileal microbiota and mucin gene expression in the growing rat.

The study was conducted to investigate whether oral administration of Korean ginseng powders can modulate gut microbiota as well as intestinal mucin production at the translational and transcriptional levels in the ileum of the growing rat. Thirty individually caged Sprague-Dawley male rats were allocated to three groups (n = 10) and fed for 21 days either a basal control diet or one of the two treatment diets each containing white or red Korean ginseng (WG or RG) powder. Bacterial DNA was extracted from ileal digesta and subjected to quantitative real-time PCR (qPCR) using primers for total bacteria, Lactobacillus, Bifidobacteria, Escherichia coli, Bacteroides, and Clostridium strains. The qPCR results showed that consumption of WG or RG powder significantly increased the number of total bacteria and Lactobacillus strains compared to the control group. Consumption of WG powder increased mRNA expression of the Muc2 gene in the small intestine compared to the control group. There was no effect of WG or RG on the small intestinal digesta mucin content. Correlation analysis showed that expression of the Muc2 gene was significantly associated with the number of total bacteria (r = 0.52, P < 0.05) and Lactobacillus strains (r = 0.53, P < 0.05), respectively. Furthermore, the number of Lactobacillus strains was significantly correlated with the number of total bacteria (r = 0.87, P < 0.05). Consumption of the WG powder modulated the intestinal ecosystem of the growing rat and intestinal mucin gene expression.

Green and gold kiwifruit peel ethanol extracts potentiate pentobarbital-induced sleep in mice via a GABAergic mechanism.

Kiwifruit is one of the most popular fruits worldwide, and it has various biological properties, including antioxidant, anti-allergic, and cardiovascular protective effects. The peel of kiwifruit, which is a by-product of processing, is a good source of flavonoids; however, its bioactivity has not been widely investigated. In this study, we evaluated the hypnotic effects of green (GRPE, Actinidia deliciosa) and gold (GOPE, Actinidia chinensis) kiwifruit peel ethanol extracts and their solvent fractions, and the possible underlying mechanisms. Oral GRPE and GOPE administration (125-1000mg/kg) produced a dose-dependent decrease in sleep latency and an increase in sleep duration in pentobarbital-treated mice. Among three different solvent fractions of GRPE and GOPE, ethyl acetate (EA) fractions had the greatest effect on sleep duration at 250mg/kg. The total flavonoid contents of solvent fractions were proportional to sleep duration. Like diazepam (a GABA(A)-benzodiazepine (BZD) receptor agonist), the hypnotic effects of GRPE, GOPE, and their EA fractions were fully inhibited by flumazenil (a GABA(A)-BZD receptor antagonist). These results suggest that potentiation effects of GRPE and GOPE on pentobarbital-induced sleep in mice may be modulated by a GABAergic mechanism.

Comparative analysis of RNA silencing suppression activities between viral suppressors and an endogenous plant RNA-dependent RNA polymerase.

RNA silencing is an evolutionarily conserved system that functions as an antiviral mechanism in eukaryotes, including higher plants. To counteract this, several plant viruses express silencing suppressors that inhibit RNA silencing in host plants. Here, we show that both 2b protein from peanut stunt virus (PSV) and a hairpin construct (designated hp-RDR6) that silences endogenous RNA-dependent RNA polymerase 6 (RDR6) strongly suppress RNA silencing. The Agrobacterium infiltration system was used to demonstrate that both PSV 2b and hp-RDR6 suppressed local RNA silencing as strongly as helper component (HC-Pro) from potato virus Y (PVY) and P19 from tomato bush stunt virus (TBSV). The 2b protein from PSV eliminated the small-interfering RNAs (siRNAs) associated with RNA silencing and prevented systemic silencing, similar to 2b protein from cucumber mosaic virus (CMV). On the other hand, hp-RDR6 suppressed RNA silencing by inhibiting the generation of secondary siRNAs. The small coat protein (SCP) of squash mosaic virus (SqMV) also displayed weak suppression activity of RNA silencing. Agrobacterium-mediated gene transfer was used to investigate whether viral silencing suppressors or hp-RDR6 enhanced accumulations of green fluorescence protein (GFP) and ß-glucuronidase (GUS) as markers of expression in leaf tissues of Nicotina benthamiana. Expression of both GFP and GUS was significantly enhanced in the presence of PSV 2b or CMV 2b, compared to no suppression or the weak SqMV SCP suppressor. Co-expression with hp-RDR6 also significantly increased the expression of GFP and GUS to levels similar to those induced by PVY HC-Pro and TBSV P19.

Ovine serum immunoglobulin has immunomodulatory effects in growing rats gavaged with Salmonella enteritidis.

In this study, we aimed to determine whether orally administered ovine serum Ig modulate aspects of immunity and associated gut microflora in growing rats challenged with Salmonella enteritidis. The 4 groups consisted of rats fed a casein-based control diet (BD; ungavaged) and 3 groups of rats gavaged with 1 × 10(7) viable Salmonella enteritidis and fed a BD diet, a BD diet containing freeze-dried ovine Ig (FDOI), or a BD diet containing inactivated ovine Ig (IOI). The rats were randomly allocated to 1 of the 4 diets (n = 15) and consumed it for 18 d. They were orally gavaged on d 15. Phagocytic activity of peripheral blood leukocyte and lymphocyte proliferation in the presence of the concanavalin A (ConA) were greater (P < 0.05) in the ungavaged BD- and gavaged FDOI-fed rats than in the gavaged rats fed either the BD or IOI diet. ConA-stimulated Peyer's patch cells and splenocytes from the gavaged rats fed the FDOI diet produced more IFNγ, IgA, and IgG than the gavaged rats fed either the BD or IOI diet (P < 0.05). The gavaged FDOI-fed rats had higher ileal and colonic digesta and plasma concentrations of anti-Salmonella secretory sIgA and secretory sIgG (P < 0.05). DNA analysis of a denatured gradient gel electrophoresis profile revealed that 6 of 10 bands had sequence similarity to probiotic strains of bacteria in the ileum and colon of the gavaged FDOI-fed rats. In conclusion, an ovine Ig fraction modulated various indices of immune function and associated gut microflora in growing rats inoculated with Salmonella.

Indian culinary plants enhance glucose-induced insulin secretion and glucose consumption in INS-1 ß-cells and 3T3-L1 adipocytes.

Six Indian plants, commonly used as culinary plants, herbs or spices (kikar; jamun; neem; harad; fenugreek; bitter gourd), were screened and compared for their antidiabetic potential in vitro. Aqueous plant extracts were prepared and assessed for their effect on the insulin secretion activity of rat pancreatic INS-1 ß-cells and glucose consumption in mouse 3T3-L1 adipocytes in order to study their specific mechanisms of action. The effect of the plant extract concentration (25-1000µg/ml) on insulin release and glucose consumption was also studied. All the extracts had a significant stimulatory effect on the insulin secretion of INS-1 cells. In the presence of kikar extract (100µg/ml), an increase of 228% in insulin release was recorded compared to the control (5.6mM glucose) whereas that was 270% and 367% in the presence of kikar and jamun extracts (500µg/ml), respectively. 3T3-L1 cells treated with jamun extract (100µg/ml) exhibited the highest increase in glucose consumption by the cells (94%, compared with the control) followed by harad (53%) and fenugreek (50%) extracts. A significant inhibitory effect of the fenugreek, kikar and jamun extracts on glucose diffusion across a dialysis membrane suggested that these extracts could partly act by decreasing glucose absorption in the small intestine. The results showed that a combination of these plants in diet could help in the management of both type 1 and type 2 diabetes.

The in vitro anti-pathogenic activity of immunoglobulin concentrates extracted from ovine blood.

An immunoglobulin-rich fraction has been prepared from ovine blood in our laboratory. We have investigated its antibacterial activity and binding activity to pathogenic whole cell antigens, lipopolysaccharide (LPS) and staphylococcal enterotoxin B. Ovine immunoglobulin concentrate (OIC) comprised about 73 +/- 2% of IgG and 11 +/- 1% of IgM on a protein basis. It inhibited the growth of all 13 strains of pathogens tested, but the inhibitory activity varied according to bacterial strain. The inhibitory activity of OIC was attributed to the high contents of undenatured immunoglobulin present because its inhibitory activity was destroyed by pepsin digestion and heat treatment (65 degrees C for 30 min). OIC bound to all the Gram-positive and Gram-negative pathogens, regardless of cell wall structure. The highest magnitude of crossreactivity to whole cell antigens was against Staphylococcus epidermidis and Shigella soneii strains (p < 0.001). The binding activity of OIC to LPS obtained from Escherichia coli O111:B4 and Salmonella enterica serotype typhimurium was assessed by enzyme-linked immunosorbent assay and lymphoblast K-562 proliferation assay. OIC bound to LPS with a binding activity that was dependent on OIC concentration and saturable, showing typical hyperbolic curves. For toxin-binding activity, an OIC concentration-dependent trend like that for LPS-binding activity was also observed. This preliminary evidence suggests that the OIC used in this study could be a promising supplement for protecting against pathogenic bacteria.