Co-sonicated coacervation for high-efficiency green nanoencapsulation of phytosterols by colloidal non-biotoxic solid lipid nanoparticles.

Plant sterols are used as a supplement or an additive to reduce LDL cholesterol. The poor dispersibility and instability of phytosterols are the main limitations of their application. So, we tried to overcome these problems through nanoencapsulation of them with colloidal natural RSs (SLNs) using an effective approach to achieve higher efficiency and less intrinsic coagulation. Phytosterols extracted from flax seeds oil with caffeine by a new method were encapsulated with a stable colloid of sheep fat and ostrich oil (1:2), soy lecithin, and glucose through co-sonicated coacervation. Characterization of the obtained SLNs was conducted using FTIR, UV-Vis, SEM, DLS, and GC analysis. The three-factor three-level Behnken design (BBD) was used to prioritize the factors affecting the coacervation process to optimize particle size and loading capacity of SLNs. Operational conditions were examined, revealing that the size of SLNs was below 100 nm, with a phytosterols content (EE %) of 85.46% with high positive zeta potential. The nanocapsules' anti-microbial activity and drug-release behavior were then evaluated using the CFU count method and Beer-Lambert's law, respectively. The controlled release of nanocapsules (below 20%) at ambient temperature has been tested. The stability of nano-encapsulated phytosterols was investigated for six months. All results show that this green optimal coacervation is a better way than conventional methods to produce stable SLNs for the nanoencapsulation of phytosterols.

Effects of a novel probiotic mixture on the modulation of brain and intestine Aquaporin-4 gene expression in rats exposed to Cadmium.

Cadmium (Cd) is a toxicant metal that risks human and animal health. Nowadays, the vital role of Aquaporin-4 (AQP-4) in brain and gut cell permeability has gathered too much attention to protecting against heavy metals. Studies have shown that heavy metals can harm the body due to oxidative stress. Probiotics are known for their health-beneficial effects and establish as dietary adjuncts mainly for their antioxidant properties. This study investigated the impact of a novel probiotic combination including Lactobacillus casei IBRC-M10783, Lactobacillus rhamnosus IBRC-M10782, and Lactobacillus helveticus TG-34 on the AQP-4 gene expression in CdCl2-induced Wistar rats. Rats were divided into three groups and received a specific dose of CdCl2 or probiotics. The AQP-4 expression level had estimated by Real-Time PCR in both the intestine and brain. These results showed a significant reduction in AQP-4 gene expression in the probiotic treatment group compared to the CdCl2 control group in the intestine and brain for the first time. Our research showed that consuming a probiotic mixture of L. casei, L. rhamnosus, and L. helveticus can reduce the expression of the aquaporin-4 gene in the brain and intestine of rats exposed to Cadmium, which can be promising in the field of aquaporin-4 regulation.

A novel probiotic strain exerts therapeutic effects on mouse model of multiple sclerosis by altering the expression of inflammasome and IDO genes and modulation of T helper cytokine profile.

Multiple sclerosis is an inflammatory demyelinating disease that commences to neuronal cell destruction. Recently, a promising evidence of synergic effects of combined supplementation with vitamin D and probiotics in modulating the gut microbiota and metabolome is emerging. Bacillus Coagulans IBRC-M10791 as a novel strain was chosen, prevention and treatment impacts of regular administered were studied in Cuprizone-induced C57bl/6 mouse of demyelination. The mice were divided into six groups and received a daily dose of cuprizone or probiotics. To investigate the effect of probiotic, the IDO-1, CYP27B1, NLRP1, NLRP3, and AIM2 expression were estimated by Real-Time PCR, and IL-4, IL-17, IFN-gamma, and TGF-beta cytokines were measured by ELISA. The results showed that there was significant decrease in IL-17 and IFN-γ and modulatory effects on IL-4 and TGF-ß. On the other hand, we demonstrated that there are significant decrease for expression of IDO-1, CYP27b1, NLRP1, NLRP3 and AIM2 genes in prevention and treatment groups compared to cuprizone group. Also, a significant enhancement in rate of remyelination and alternations proved by LFB staining and Y-Maze test. In conclusion, our study provides insight into how the therapeutic effect of the chosen strain of probiotic was correlated with the modulation of the level of inflammatory and anti-inflammatory cytokines. Further, we demonstrated that the expression of genes related to Tryptophan, Vitamin D and Inflammasome pathways could be affected by B.coagulans. Our study could be beneficial to provide a novel Co-therapeutic strategy for Multiple sclerosis.

Analysis of inflammasomes and CYP27B1 genes in cuprizone demyelinated C57BL/6 mice and evaluation of Th1 and Th2 patterns after oral administration of Lactobacillus casei strain T2 (IBRC-M10783).

Multiple sclerosis is characterized by the destruction of myelin in the CNS. Various factors including genetics, epigenetics, and environmental factors are involved in the development of the disease. There is evidence that changes in the gut microbiome profile are associated with immune-related diseases such as MS. Probiotics can alter the composition of the gut microbiota on the mucosal surfaces by differentiating naive T cells into Th1, Th2, Th17, and Treg cells. Female C57BL/6 mice were divided into 6 groups (n = 7): Normal group, cuprizone group (gavage of cuprizone for 4 weeks), Probiotic group (gavage of probiotic for 4 weeks), Treatment1 group (Probiotic for 4 weeks and then cuprizone for 4 weeks), treatment2 group (cuprizone for 4 weeks and then probiotic for 4 weeks) and treatment3 group (cuprizone for 4 weeks and then probiotic for 4 weeks with vitamin D3). Then the expression of NLRP-1, NLRP-3, AIM2, and CYP27B1 genes were evaluated using Real-Time PCR, and serum levels of IFN-γ and IL-4 were also measured by ELISA.The results showed a significant decrease in the expression of inflammasome and CYP27B1 genes in the probiotic-treated groups compared to the cuprizone group. Also, the comparison between probiotic-treated groups and cuprizone group showed a significant decrease in the amount of IFN-γ and IL-4. Due to reduced expression of the inflammasome genes as well as the decrease in IFN-γ levels as an inflammatory cytokine, it appears that L. casei may be effective in the healing process of demyelinated mice.

Effects of a Potential Probiotic Strain Lactobacillus gasseri ATCC 33323 on Helicobacter pylori-Induced Inflammatory Response and Gene Expression in Coinfected Gastric Epithelial Cells.

In the present study, we aimed to investigate the modulatory effects of a potential probiotic bacterium Lactobacillus gasseri ATCC 33323 on Helicobacter pylori-induced inflammatory response and gene expression in human gastric adenocarcinoma (AGS) cell line. The gastric epithelial cells were coinfected with a collection of H. pylori clinical strains alone or in combination with L. gasseri at a multiplicity of infection (MOI) of 1:100 for each bacterium, and incubated for different time points of 3, 6, and 12 h. IL-8 secretion from coinfected AGS cells after incubation at each time point was measured by an enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-8, Bcl-2, ß-catenin, integrin α5, and integrin ß1 genes was determined by quantitative RT-PCR amplification of total RNA extracted from coinfected epithelial cells. L. gasseri significantly (P < 0.05 and P < 0.01) decreased the production of IL-8 in AGS cells coinfected with H. pylori strains at 6 h post-infection. We also detected that L. gasseri significantly (P < 0.05) down-regulated the gene expression level of IL-8 in H. pylori-stimulated AGS cells after 6 and 12 h of coinfection. Similarly, L. gasseri caused a significant decrease (P < 0.05) in mRNA expression of Bcl-2, ß-catenin, integrin α5, and integrin ß1 genes in AGS cells at 3 and 6 h after infection with H. pylori strains as compared with non-infected control cells. In conclusion, our results demonstrated that L. gasseri ameliorates H. pylori-induced inflammation and could be developed as a supplementation to the current treatment regimens administrated against H. pylori infection.

Effects of probiotic lactic acid bacteria on growth performance, carcass characteristics, hematological indices, humoral immunity, and IGF-I gene expression in broiler chicken.

The effects of screened lactic acid bacteria strains were evaluated on growth performance, humoral immunity, and IGF-1 gene expression in broiler chickens. The three dietary groups of negative control fed basal diet, the native LAB probiotic group (NP), and PrimaLac commercial LAB probiotic (PC) were studied. The results revealed that NP and PC diets significantly improved feed conversion ratio and increased body weight, as well as relative weight of carcass compared with group fed NC diet (P < 0.05). Lymphocyte level was significantly increased in birds fed NP and PC (P < 0.01), while serum triglycerides and total cholesterol levels were significantly decreased compared with the NC (P < 0.05). Significant increases were observed in antibody titers against Newcastle disease virus of vaccinated birds (P < 0.03), and morphological analysis of ileum revealed significant increases (P < 0.05) in the villus height and villus height/crypt depth in birds fed NP and PC compared with the NC. The dietary significantly increased Lactobacillus spp. (P < 0.05), while Escherichia coli (P < 0.04) populations were significantly decreased, and also, the expression of IGF-1 gene in liver tissue of broilers fed NP and PC was significantly increased compared with the NC (P < 0.05). These results indicated that the identified native LAB strains can be used commercially as a low-cost probiotic in poultry industry of Iran.

The effects of probiotics and synbiotic supplementation on glucose and insulin metabolism in adults with prediabetes: a double-blind randomized clinical trial.

AIMS: Probiotics and/or prebiotics could be a promising approach to improve metabolic disorders by favorably modifying the gut microbial composition. OBJECTIVES: To assess the effects of probiotics and synbiotic on glycemic indices in prediabetic individuals who are at risk of type 2 diabetes and its complications. METHODS: In a double-blind, randomized, placebo-controlled parallel-group clinical trial, 120 prediabetic adults participated and were randomly allocated to receive either probiotics or synbiotic or placebo supplements for 24 weeks. Anthropometric measurements, food record, physical activity and glycemic biomarkers including glycated hemoglobin (HbA1C), fasting plasma glucose (FPG), fasting insulin levels (FIL), homoeostasis model assessment for insulin resistance (HOMA-IR), quantitative insulin sensitivity check index (QUICKI), and ß-cell function (HOMA-B) were assessed at baseline and repeated at 12 and 24 weeks and compared within and between three groups using repeated measure ANOVA. RESULTS: Compared with the placebo, synbiotic supplementation resulted in a higher significant reduction in FPG (- 6.5 ± 1.6 vs. - 0.82 ± 1.7 mg/dL, P = 0.01), FIL (- 2.6 ± 0.9 vs. - 0.8 ± 0.8 µIU/mL, P = 0.028), and HOMA-IR (- 0.86 ± 0.3 vs. - 0.16 ± 0.25, P = 0.007), and a significant elevation in the QUICKI (+ 0.01 ± 0.003 vs. + 0.003 ± 0.002, P = 0.006). In addition, significant decreases in HbA1C was seen following the supplementation of probiotics and synbiotic compared with the placebo (- 0.12 ± 0.06 and - 0.14 ± 0.05 vs. +0.07 ± 0.06%, P = 0.005 and 0.008, respectively). HOMA-B was not found to be different between or within the three groups. CONCLUSION: Glycemic improvement by probiotics and particularly synbiotic supplements in prediabetic individuals has been supported by current study. However, further studies are required for optimal recommendations in this important area of patient treatment. TRIAL REGISTRATION: Iranian Registry of Clinical Trials: IRCT201511032321N2, Date registered February 27, 2016.

Antimutagenic and anticancer effects of lactic acid bacteria isolated from Tarhana through Ames test and phylogenetic analysis by 16S rDNA.

Deaths due to cancer have been increasing in many countries in recent years. One of the beneficial probiotic properties is potential of antimutagenic and anticancer effects. The aim of this study was to investigate antimutagenic and anticancer effects of 25 strains of lactobacilli spp isolated from Tarhana in vitro conditions and to identify selected strains based on molecular biology. Antimutagenicity and anticarcinogenicity tests were performed based on proposed method of Professor Ames et al. Identification of selected strains was performed based on biochemical and molecular tests. Study of antimutagenic and anticancer effects of 25 different strains led to identification of 4 isolates of Lactobacillus spp. Higher antimutagenic activities against sodium azide were observed in cell suspensions of 4 strains as compared with their supernatants. So the inhibitory percentage of cell suspensions among 4 strains was equal to 60.38% in its highest level. Antimutagenicity of these strains was also increased in the presence of rat S9. Four selected strains were identified based on 16S rDNA sequence. Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus brevis isolated from Tarhana exhibited high antimutagenic and anticancer activity. Of 25 strains of lactobacilli, 4 strains with the highest antimutagenicity effect were chosen as new potential probiotic strains.

Plasmin digest of κ-casein as a source of antibacterial peptides.

This study investigated the antibacterial properties of plasmin, the plasmin hydrolysis of bovine κ-casein and the fractions (named κC1, κC2, κC3, κC4, and κC5) liberated from it using RP-HPLC. The target bacteria were Escherichia coli, Staphylococcus aureus (pathogenic), Lactobacillus casei and Lactobacillus acidophilus (probiotic). Three peptides (kC1, kC3, and kC4) were found to have antibacterial activity, with κC3 peptide being the most active. The plasmin digest of bovine κ-casein proved to be stronger than any of its fractions in terms of antibacterial potential. Measurement of the minimum inhibitory concentration (MIC) showed that Gram-positive bacteria are generally more sensitive to antibacterial activity than Gram-negative bacteria. The MIC of nisin, as a bacteriocin peptide, was also measured. The three antibacterial peptides were identified using LC-Mass. The molecular mass of kC1, kC3, and kC4 corresponded to the f(17-21), f(22-24), and f(1-3) of bovine κ-casein, respectively. It was also found that the positive charge and hydrophobicity of a peptide are not key factors in antibacterial activity. On the whole, the present study demonstrated that the plasmin digest of κ-casein has a high antibacterial potential and can be considered as a natural antibacterial agent in the food chain.