Oral Probiotic Microcapsule Formulation Ameliorates Non-Alcoholic Fatty Liver Disease in Bio F1B Golden Syrian Hamsters

The beneficial effect of a microencapsulated feruloyl esterase producing Lactobacillus fermentum ATCC 11976 formulation for use in non-alcoholic fatty liver disease (NAFLD) was investigated. For which Bio F₁B Golden Syrian hamsters were fed a methionine deficient/choline devoid diet to induce non-alcoholic fatty liver disease. Results, for the first time, show significant clinical benefits in experimental animals. Examination of lipids show that concentrations of hepatic free cholesterol, esterified cholesterol, triglycerides and phospholipids were significantly lowered in treated animals. In addition, serum total cholesterol, triglycerides, uric acid and insulin resistance were found to decrease in treated animals. Liver histology evaluations showed reduced fat deposits. Western blot analysis shows significant differences in expression levels of key liver enzymes in treated animals. In conclusion, these findings suggest the excellent potential of using an oral probiotic formulation to ameliorate NAFLD.     

Design and validation of an orally administrated active L. fermentum-L. acidophilus probiotic formulation using colorectal cancer Apc Min/+ mouse model

Probiotics have been shown to have beneficial properties in attenuating the risk of colorectal cancer (CRC) development. However, functional evidence to support such effects for some probiotic bacteria are relatively unknown. Here, we document a significant antioxidant, anti-proliferative and pro-apoptotic activities of Lactobacillus acidophilus ATCC 314 and Lactobacillus fermentum NCIMB 5221 on CRC cells, particularly when used in combination (La-Lf). Furthermore, a superior synergistic activity on the inhibition of tumor growth and modulation of cell proliferation and epithelial markers in the Apc ^Min/+ CRC mouse model was explored, based on the expression levels of Ki-67, E-cadherin, β-catenin, and cleaved caspase-3 (CC3) proteins. The anti-cancer activity of La-Lf co-culture was significantly enhanced in vitro with significant reduced proliferation (38.8 ± 6.9 %, P = 0.009) and increased apoptosis (413 RUL, P < 0.001) towards cancer cells, as well as significant protection of normal colon cell growth from toxic treatment (18.6 ± 9.8 %, P = 0.001). La-Lf formulation (10¹⁰cfu/animal/day) altered aspects of intestinal tumorigenesis by significantly reducing intestinal tumor multiplicity (1.7-fold, P = 0.016) and downregulating cellular proliferation markers, including β-catenin (P = 0.041) and Ki-67 (P = 0.008). In conclusion, La-Lf showed greater protection against intestinal tumorigenesis supporting a potential use as a biotherapeutic for the prevention of CRC.

     

An In Vitro Study of the Probiotic Potential of a Bile-Salt-Hydrolyzing Lactobacillus fermentum Strain, and Determination of Its Cholesterol-Lowering Properties

This study evaluated the use of a bile-salt-hydrolyzing Lactobacillus fermentum strain as a probiotic with potential hypocholesterolemic properties. The effect of L. fermentum on representative microbial populations and overall metabolic activity of the human intestinal microbiota was investigated using a three-stage continuous culture system. Also, the use of galactooligosaccharides as a prebiotic to enhance growth and/or activity of the Lactobacillus strain was evaluated. Administration of L. fermentum resulted in a decrease in the overall bifidobacterial population (ca. 1 log unit). In the in vitro system, no significant changes were observed in the total bacterial, Lactobacillus, Bacteroides, and clostridial populations through L. fermentum supplementation. Acetate production decreased by 9 to 27%, while the propionate and butyrate concentrations increased considerably (50 to 90% and 52 to 157%, respectively). A general, although lesser, increase in the production of lactate was observed with the administration of the L. fermentum strain. Supplementation of the prebiotic to the culture medium did not cause statistically significant changes in either the numbers or the activity of the microbiota, although an increase in the butyrate production was seen (29 to 39%). Results from this in vitro study suggest that L. fermentum KC5b is a candidate probiotic which may affect cholesterol metabolism. The short-chain fatty acid concentrations, specifically the molar proportion of propionate and/or bile salt deconjugation, are probably the major mechanism involved in the purported cholesterol-lowering properties of this strain.     

Correlation of Hair Mineral Concentrations with Insulin Resistance in Korean Males

Calcium and magnesium that are associated with insulin resistance play an antagonistic role with each other in cells. This study was conducted to investigate the relationship between hair mineral concentrations and insulin resistance in Korean adult males. A total of 123 male subjects (63 patients with metabolic syndrome and 60 normal control patients) were recruited and fasting plasma glucose, total cholesterol, triglyceride, as well as HDL cholesterol levels, HOMA-IR, and hair mineral concentrations were measured. The ratio of calcium/magnesium in hair showed a significantly positive correlation with the HOMA-IR (r?=?0.191, P?=?0.038) and insulin (r?=?0.198, P?=?0.031). The result of multiple regression analysis after adjusting the age also showed a significant correlation of the Ca/Mg ratio with HOMA-IR (R ²?=?0.115, P?=?0.047). The hair chromium concentration was lower in the metabolic syndrome group than in the control group, and it showed a significantly negative correlation with the fasting blood glucoseand the triglyceride. The result of this study showed that insulin resistance increased as the ratio of Ca/Mg increased, or as the chromium concentration in hair decreased.     

An antioxidant probiotic reduces postprandial lipemia and oxidative stress

Reducing postprandial oxidative stress (OxS), decreasing postprandial blood triglyceride level (TG) and improving lipoprotein status is likely to have a preventive impact on the development of cardiovascular disease (CVD). Previously we have shown that the antioxidant probiotic Lactobacillus fermentum ME-3 (DSM14241) is characterized by antiatherogenic effects. This randomized double-blind placebo-controlled study evaluated the influence of kefir enriched with an antioxidative probiotic L. fermentum ME-3 (LfKef) on postprandial OxS, blood TG response and lipoprotein status. 100 clinically healthy subjects were recruited into the study. Blood parameters of postprandial OxS, TG and lipoprotein status were determined by oxidized LDL, baseline diene conjugation in LDL (BDC-LDL), oxidized LDL complex with beta-2 glycoprotein (Beta2-GPI-oxLDL), paraoxonase (PON) activity, LDL-Chol, HDL-Chol and TG. To evaluate general body postprandial OxS-load we measured 8-isoprostanes (8-EPI) in the urine. Consumption of LfKef significantly reduced the postprandial level of oxidized LDL, BDC-LDL, Beta2-GPI-oxLDL, urinary 8-isoprostanes and postprandial TG and caused a significant increase in HDL-Chol and PON activity. This is the first evidence that kefir enriched with an antioxidant probiotic may have a positive effect on both postprandial OxS and TG response as well as on lipoprotein status.     

Protein hydrolysate from organic fraction of municipal solid waste compost as nitrogen source to produce lactic acid by Lactobacillus fermentum ATCC 9338 and Lactobacillus plantarum NCIMB 8826

In this work a strategy for obtaining free amino-acids concentrate from an organic fraction of municipal solid waste compost and its use as a nitrogen source for lactic acid production, a compound widely used in different industries, using L. fermentum ATCC 9338 and L. plantarum NCIMB 8826 strains is described. Enzymatic digestion is based on the combined action of endoprotease Alcalase 1.5 MG and exoprotease Flavourzyme 500 MG. The highest degree of hydrolysis obtained under the optimal conditions was 41%. The use of glucanase Viscozyme L prior to protein hydrolysis helped to reduce the viscosity of the solution and promote the action of proteases, increasing its hydrolysis degree by 76%. The hydrolysate contained all 21 amino-acids, making it ideal for lactic acid bacteria growth. During shake flask cultivations the culture media was complemented with glucose as carbon source. Finally, with the hydrolysate, a maximum lactic acid concentration of 9.0 ± 0.2 g·L⁻¹ and 11.1 ± 0.1 g·L⁻¹ for L. fermentum ATCC 9338 and L. plantarum NCIMB 8826 respectively was obtained after 27 h. The innovation of the approach lies in exploiting the overproduction of compost for the production of lactic acid.     

Association of Increased Hair Calcium Levels and Enhanced Augmentation Index (AIx): a Marker of Arterial Stiffness

Arterial stiffness is involved in the pathophysiology of cardiovascular disease, and the degree of arterial stiffness is associated with the extent of vascular calcification. This study aimed to investigate the association of hair calcium levels with augmentation index (AIx), a simple, non-invasive measurement for arterial stiffness. Healthy Koreans (male, n?=?34, female, n?=?70) were enrolled in this study. Anthropometric parameters, lipid profiles, fasting glucose, hair mineral levels, and AIx were measured. Pearson/partial correlations and multivariate linear regression analyses were used to assess the relationship between hair calcium levels and AIx. AIx positively correlated with hair calcium levels (r?=?0.275, p?=?0.005), age (r?=?0.283, p?=?0.004), systolic blood pressure (r?=?0.282, p?=?0.004), low-density lipoprotein (LDL)-cholesterol (r?=?0.255, p?=?0.009), and hair magnesium (r?=?0.196, p?=?0.046), and negatively correlated with heart rate (r?=?-0.563, p?<?0.001) and fasting glucose (r?=?-0.262, p?=?0.005). Hair calcium levels significantly correlated with hair magnesium (r?=?0.926, p?<?0.001). Significant relationship between AIx and hair calcium levels was maintained after adjustment for sex, age, height, hear rate, blood pressure, total cholesterol, LDL-cholesterol, fasting glucose, and hair magnesium (r?=?0.244, p?=?0.018). Logistic regression model showed that AIx increased with the increment of hair calcium levels; log-AIx increased by 0.403% (95% CI: 0.139–0.515, p?=?0.001) per unit change in log-hair calcium level (sex-adjusted). After adjustment for all the variables above together with triglyceride and high-density lipoprotein-cholesterol, increased significance of the association was 0.513% (p?=?0.016)]. This study supports the presence of the independent positive relationship between hair calcium levels and AIx. It suggests the possibility that hair calcium levels may be a useful index for reflecting arterial stiffness.     

人源乳酸菌对2型糖尿病小鼠的缓解效果

为考察人源乳酸菌(发酵乳杆菌Lactobacillus fermentum 11、305和植物乳杆菌Lactobacillus plantarum 22、25)对2型糖尿病的缓解效果,对2型糖尿病小鼠连续灌胃菌粉溶液12周.每周记录各组小鼠体重、进食量和血糖.实验结束前进行口服葡萄糖耐量试验和胰岛素抵抗试验.实验结束后...     

Interaction of Zinc, Ascorbic Acid, and Folic Acid in Glycation with Albumin as Protein Model

Using albumin as model, we conducted series of in vitro glycation experiments to examine role of zinc in glycation using glucose at 4–100 mg/ml, incubations at 37°C or 60°C, duration of 2 or 4 weeks and in presence of zinc or ascorbic acid (AA) or folic acid (FA). Modifications of bovine serum albumin (BSA) were examined by using fluorescence of advanced glycation end products (AGEs) and dityrosine, UV, and Fourier transformed infrared spectroscopy. Adding zinc (0 to 768.5 μmol/l) resulted in significant inhibition of albumin glycation by glucose with a linear fit, $ y = - 0.0{895}x + {23}0.{99}\left( {{R^2} = 0.{7676},p = 0.0{13}} \right) $ . The glycation by fructose was greater than that of glucose with stronger inhibitory effect by zinc in fructose–glycation (t?=??5.8, p?=?0.002). Addition of zinc significantly decreased fluorescence as seen in Zn?+?FA or Zn?+?AA sets as compared to sets of FA alone (p?=?0.00056) or AA alone (p?=?0.037). The fluorescence for dityrosine and AGE had a correlation of 0.897 (p?<?0.01). The data from fluorescence, UV, and FTIR spectra collectively suggested inhibitory effect of zinc in BSA glycation alone or in presence of FA and AA, showing new dimension for the protective action of zinc in hyperglycemic conditions.     

Relationships of Hair Mineral Concentrations with Insulin Resistance in Metabolic Syndrome

Mineral deficiencies can cause impaired insulin release and insulin resistance. This study was conducted to investigate the relationship between hair mineral concentrations and insulin resistance in patients with metabolic syndrome (MS). A total of 456 subjects (161 patients with MS and 295 subjects without MS) were reviewed, and fasting plasma glucose, triglycerides, HDL-cholesterol, homeostasis assessment model-insulin resistance (HOMA-IR), and hair mineral concentrations were analyzed. While hair sodium and potassium concentrations were significantly higher, the hair calcium, magnesium, and zinc concentrations were lower in the MS group than in the control group. Regarding toxic element measurements, the hair arsenic (As) and lead (Pb) concentrations were higher in the MS group than in the control group. The results of multiple regression analysis, after adjusting for age, showed significant relationships between the Na/Mg and Ca/P ratios and HOMA-IR (R ²?=?0.109, p?<?0.05). The Ca, Na, K, and B concentrations were also associated with HOMA-IR (R ²?=?0.116, p?<?0.05). The hair Na concentration was significantly associated with MS, even after adjusting for age, visceral adipose tissue, and HOMA-IR (OR 1.020; 95 % CI 1.001–1.040; p?=?0.036). Our findings suggest that hair mineral concentrations, such as calcium, magnesium, zinc, sodium, and potassium concentrations, may play a role in the development of insulin resistance.     

Behavior of Lactobacilli Isolated from Fermented Slurry (ben-saalga) in Gnotobiotic Rats

Most bacterial strains, which have been studied so far for their probiotic functions, are extensively used by manufacturers in developed countries. In our work, we sought to study a mix (called BSL) comprising three strains belonging to Lactobacillus fermentum, L. paraplantarum and L. salivarius, that were isolated from a traditional African pearl millet based fermented slurry. Our objective was to study this BSL cocktail in gnotobiotic rats, to evaluate their survival and their behavior in the digestive tract conditions. After a single oral inoculation of germfree rats with BSL, the species established stably in the digestive tract with the following hierarchy of abundance: L. salivarius> L. plantarum> L. fermentum. BSL cocktail was metabolically active since it produced 50 mM lactate and it expressed genes involved in binding mechanism in the caecum. Furthermore, the global morphology of the colon epithelium was not disturbed by the BSL cocktail. BSL cocktail did not modify mucus content and host mucus-related genes (MUC1, MUC2, MUC3 or resistin-like molecule β). The cocktail of lactobacilli enhanced the proliferating cell nuclear antigen (PCNA) at a level comparable to what was observed in conventional rats. PCNA was involved in proliferation and DNA repair, but the presence of the cocktail did not provoke proliferative events (with Ki67 as indicator), so we suppose BSL may help gut preservation. This work is the first step towards the selection of strains that are derived from traditional fermented food to formulate new probiotic mixture.     

Short-term weight loss in overweight/obese low-income women improves plasma zinc and metabolic syndrome risk factors

Metabolic syndrome is a group of disorders involving obesity, insulin resistance, dyslipidemia and hypertension. Obesity is the most crucial risk factor of metabolic syndrome, because it is known to precede other risk factors. Obesity is also associated with disturbances in the metabolism of the trace mineral, zinc. The overall purpose of this study was to investigate the effects of short-term weight loss on plasma zinc and metabolic syndrome risk factors. An 8-week weight loss intervention study was conducted with 90 low-income overweight/obese mothers, whose youngest child was 1–3 years old. Plasma levels of zinc, glucose, insulin, leptin, triglycerides, total, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) cholesterol were measured and compared at weeks 0 and 8 of the weight loss program. At pre-study, plasma zinc was low in 39% and, within normal values in 46%, of obese/overweight mothers. By the end of intervention, plasma zinc rose by 22% and only 5% of the mothers continued to exhibit low plasma zinc. At post-study, the metabolic syndrome risk factors of waist circumference, HDL cholesterol, and diastolic blood pressure (p < 0.05) showed significant improvements. Plasma zinc increased by a greater margin (67%) in women with low zinc, as compared to those with normal zinc (18%); weight reduction was similar in both the groups. Finally, changes in % body fat were related negatively with changes in plasma zinc (r = ? 0.28, p < 0.05). The circulating levels of zinc, as well as the metabolic syndrome components, showed significant improvements in overweight/obese low-income women after weight loss.     

Fusaric Acid-Producing Strains of Fusarium oxysporum Alter 2,4-Diacetylphloroglucinol Biosynthetic Gene Expression in Pseudomonas fluorescens CHA0 In Vitro and in the Rhizosphere of Wheat

The phytotoxic pathogenicity factor fusaric acid (FA) represses the production of 2,4-diacetylphloroglucinol (DAPG), a key factor in the antimicrobial activity of the biocontrol strain Pseudomonas fluorescens CHA0. FA production by 12 Fusarium oxysporum strains varied substantially. We measured the effect of FA production on expression of the phlACBDE biosynthetic operon of strain CHA0 in culture media and in the wheat rhizosphere by using a translational phlA′-′lacZ fusion. Only FA-producing F. oxysporum strains could suppress DAPG production in strain CHA0, and the FA concentration was strongly correlated with the degree of phlA repression. The repressing effect of FA on phlA′-′lacZ expression was abolished in a mutant that lacked the DAPG pathway-specific repressor PhlF. One FA-producing strain (798) and one nonproducing strain (242) of F. oxysporum were tested for their influence on phlA expression in CHA0 in the rhizosphere of wheat in a gnotobiotic system containing a sand and clay mineral-based artificial soil. F. oxysporum strain 798 (FA⁺) repressed phlA expression in CHA0 significantly, whereas strain 242 (FA⁻) did not. In the phlF mutant CHA638, phlA expression was not altered by the presence of either F. oxysporum strain 242 or 798. phlA expression levels were seven to eight times higher in strain CHA638 than in the wild-type CHA0, indicating that PhlF limits phlA expression in the wheat rhizosphere.     

Effect of Soymilk Fermented with Lactobacillus plantarum P-8 on Lipid Metabolism and Fecal Microbiota in Experimental Hyperlipidemic Rats

We recently identified a novel probiotic strain Lactobacillus plantarum P-8 (L. plantarum P-8), which has been characterized in detail with regard to its probiotic potential. In the present study, soymilk fermented with L. plantarum P-8 was examined for its effects on diet-induced hyperlipidemia in Wistar rats. The experimental animals were divided into four groups: control group (C group), model group (M group), soymilk group (SM group) and fermented soymilk group (FSM group). The serum lipid levels, hepatic fat deposition, serum oxidative stress parameters, hepatic marker enzymes levels, organ indices, gut bacteria and fecal fat contents were analyzed. Fermented soymilk reduced the concentration of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in serum, with a significant elevation in high-density lipoprotein cholesterol (HDL) concentration. Our results also suggested the beneficial effects of fermented soymilk on the liver function, hyperlipidemia-induced oxidative stress and intestinal bacteria. Moreover, fermented soymilk could enhance the fecal excretion of TC, TG and bile acids. These findings demonstrated that soymilk fermented with L. plantarum P-8 was effective in improving the lipid metabolism in hyperlipidemic rats. The hypolipidemic effect of fermented soymilk was partly due to the inhibition of dietary fats absorption and regulation of fecal fats excretion mediated by gut bacteria.     

Altered Trace Mineral Milieu Might Play An Aetiological Role in the Pathogenesis of Polycystic Ovary Syndrome

Insulin resistance is a very common associate of polycystic ovary syndrome (PCOS). Pathophysiology in relation with the essential elements including copper, magnesium, zinc, manganese, chromium, and calcium has been reported in women with insulin resistance. This prospective study was designed to explore whether the women with PCOS do exhibit altered serum element levels in association with/without insulin resistance. One hundred and thirty-two women with PCOS and forty-six control women were studied. Women with PCOS were further divided based on the presence of insulin resistance (insulin resistant: n?=?50; non-insulin resistant: n?=?82). In all women, basal levels of gonadotropins, prolactin, testosterone, insulin, glucose, and the six different elements were measured. Serum levels of testosterone (p?<?0.001), luteinizing hormone (p?<?0.05), and fasting insulin (p?<?0.004) were significantly higher in the PCOS population compared to controls as well as PCOS women without insulin resistance. Women with PCOS exhibited a significantly high calcium (p?<?0.04) and lower manganese levels (p?<?0.002) when compared to controls. However, the PCOS women with insulin resistance exhibited significantly lower serum levels of magnesium and chromium (p?<?0.04), in addition to higher levels of zinc and copper (p?<?0.04). The differences in calcium (p?<?0.03) and manganese levels (p?<?0.0001) became aggravated with the presence of insulin resistance when compared to control as well as PCOS women without insulin resistance. In PCOS-associated insulin resistance, circulating serum magnesium (r?=??0.31; p?<?0.03) and chromium (r?=??0.38; p?<?0.006) status significantly correlated with fasting insulin levels. We conclude that imbalanced element status may be a key foundation for insulin resistance in PCOS. The findings in this study should be investigated with further trials in order to obtain new insights into PCOS.     

Screening of potential probiotic lactic acid bacteria and production of amylase and its partial purification

Probiotics are the healthy living bacteria when administered in adequate amounts confers health benefits in the host. The main objective of present study was to screen the bacteria for potential probiotic characters and enzyme production. The probiotic characters like tolerance to low pH, bile salts, antibiotic sensitivity, hydrophobicity and auto-aggregation properties were evaluated. Among all isolates Lactobacillus fermentum and Lactobacillus sp G3_4_1TO2 showed maximum potential probiotic characters and produced amylase enzyme by observing the halo zone around the colonies with the diameter 0.9?mm and 1.23?mm. Lactobacillus sp G3_4_1TO2 produced maximum amylase when compared with Lb. fermentum. The protein yield was 55.4% with the specific activity of 88.9 U/mg and obtained 40.8% purification fold. The molecular weight of amylase enzyme determined by SDS PAGE was 95,000?Da. From the present study it was considered that Lactobacillus sp G3_4_1TO2 was a potential probiotic bacteria producing maximum amylase enzyme.     

Selenium, Zinc, and Copper Plasma Levels in Patients with Schizophrenia: Relationship with Metabolic Risk Factors

The aim of this study was to determine the plasma selenium (Se), copper (Cu), and zinc (Zn) levels and to evaluate their possible association with metabolic syndrome (MetS) components in patients with schizophrenia. The study group consisted of 60 patients with schizophrenia and 60 sex- and age-matched healthy controls. Anthropometric measurements, blood pressure, and biochemical analysis of fasting blood were performed in all subjects. Patients with schizophrenia had significantly higher plasma Cu concentrations compared with controls (0.97?±?0.31 vs. 0.77?±?0.32 mg/L, p?=?0.001). The plasma Cu concentration showed a positive correlation with plasma glucose and diastolic blood pressure in the patient groups (r _s ?=?0.263, p?<?0.05 and r _s ?=?0.272, p?<?0.05, respectively). The plasma Se level correlated positive with MetS score (r _s ?=?0.385, p?<?0.01), waist circumference (r _s ?=?0.344, p?<?0.05), plasma glucose (r _s ?=?0.319, p?<?0.05), and triglyceride concentrations (r _s ?=?0.462, p?<?0.001) in patients with schizophrenia. Plasma Zn did not correlate with any of the MetS components. These results suggest that alterations in plasma Cu and Se levels in medicated patients with schizophrenia could be associated with metabolic risk factors.     

人生殖道源乳酸杆菌的筛选及在小鼠阴道假丝酵母病模型中的应用评价

【目的】分离筛选人阴道环境中具有益生特性的乳酸杆菌,探索外阴阴道假丝酵母病的益生菌疗法。【方法】利用含1%碳酸钙的de Man,Rogosa and Sharpe (MRS)培养基从无症状育龄女性阴道分泌物中分离乳酸杆菌,采用共培养方法评价其对白色念珠菌(Candida albicans)的抑制作用,通过对乳酸杆菌的耐酸性能、体外聚集特性和黏附能力测试考察其益生特性,并进行乳酸杆菌株功能化组合。通过构建小鼠外阴阴道假丝酵母病模型,初步探索乳酸杆菌株组合对C. albicans的抑制作用。【结果】从53个样品中分离得到19株乳酸杆菌,筛选获得4株乳酸杆菌(Lactobacillus crispatus ZH08、L. fermentumZH09、L. fermentum ZH11和L. crispatus ZH17)具有较强抑制C. albicans生长的能力。4株乳酸杆菌均能耐受低pH环境,能快速降低培养液pH。其中2株L. fermentum具有更强的抑制活性,能在24 h内快速抑制C. albicans生长,抑制率可达到95%以上;另2株L. crispatus具有更强的聚集特性和...     

Plasma diacylglycerol composition is a biomarker of metabolic syndrome onset in rhesus monkeys

Metabolic syndrome is linked with obesity and is often first identified clinically by elevated BMI and elevated levels of fasting blood glucose that are generally secondary to insulin resistance. Using the highly translatable rhesus monkey (Macaca mulatta) model, we asked if metabolic syndrome risk could be identified earlier. The study involved 16 overweight but healthy, euglycemic monkeys, one-half of which spontaneously developed metabolic syndrome over the course of 2 years while the other half remained healthy. We conducted a series of biometric and plasma measures focusing on adiposity, lipid metabolism, and adipose tissue-derived hormones, which led to a diagnosis of metabolic syndrome in the insulin-resistant animals. Plasma fatty acid composition was determined by gas chromatography for cholesteryl ester, FFA, diacylglycerol (DAG), phospholipid, and triacylglycerol lipid classes; plasma lipoprotein profiles were generated by NMR; and circulating levels of adipose-derived signaling peptides were determined by ELISA. We identified biomarker models including a DAG model, two lipoprotein models, and a multiterm model that includes the adipose-derived peptide adiponectin. Correlations among circulating lipids and lipoproteins revealed shifts in lipid metabolism during disease development. We propose that lipid profiling may be valuable for early metabolic syndrome detection in a clinical setting.     

Molecular cloning, characterization and heterologous expression of bile salt hydrolase (Bsh) from Lactobacillus fermentum NCDO394

Bile salt hydrolase (Bsh) active probiotic strains hydrolyze bile acid amino conjugates in vivo, which triggers cholesterol consumption in liver to synthesize new bile leading to consequential cholesterol lowering. Hence, bile salt hydrolyzing potential was the criterion to select L. fermentum NCDO394 for this study and its gene encoding Bsh was identified and cloned. The resulting nucleotide sequence of bsh gene contained an open reading frame (ORF) of 978 nucleotides encoding a predicted protein of 325 amino acids with a theoretical pI of 6.39. Moreover, deduced Bsh protein had high similarity with the Bshs of L. fermentum only and also exhibited significant similarity to the Pencillin V amidases of other Lactobacillus spp. Five catalytically important amino acids were highly conserved in L. fermentum Bsh while four amino acid motifs around these active sites, were not as consistent as in other Bsh proteins. Furthermore, L. fermentum bsh gene was sub-cloned into pET-28b(+) vector, and its expression was induced with 0.05 mM isopropylthiogalactopyranoside (IPTG) in Escherichia coli BL21(DE3). The recombinant Bsh (rBsh) was purified with homogeneity using Ni⁺²-NTA column and characterized for substrate specificity, pH and temperature. The rBsh hydrolyzed six major human bile salts with a slight preference towards glycine-conjugated bile salts. The optimum pH of rBsh was six, and its enzymatic activity declined below pH 5 and above pH 7. The enzyme was stable and functional even at 65 °C while showed its maximum activity at 37 °C. In conclusion, L. fermentum NCDO394 may be a promising candidate probiotic which may affect cholesterol metabolism in vivo.