Adjunctive role of Q10 with Ligilactobacillus salivarius, and Lactiplantibacillus plantarum probiotic Bacteria on the HEp-2 cells viability and adhesion of Streptococcus mutans.

Objective: Various studies have indicated the application of Coenzyme Q10 and probiotic bacteria such as Ligilactobacillus salivarius (L. salivarius) and Lactiplantibacillus plantarum (L. plantarum) in combating periodontal disease. Considering the positive effect of these two on oral health, and the destructive effect of S. mutans, in this study, we investigate the outcomes of the administration of probiotics and Q10 on infected HEp-2 cell viability and S. mutans adhesion in different settings. Methods: A 3-week-old human epidermoid laryngeal (HEp-2) cell line was cultured and exposed to two different probiotics and 3 different doses of Q10 doses. Samples were contaminated by S. mutans immediately (therapeutic setting) and after 3 hours (preventive setting). Eventually, the viability of HEp-2 cells was investigated by MTT. Also, the number of adhered S. mutans was explored by direct and indirect adhesion assays. Results: L. plantarum and L. salivarius protect epithelial cells against S. mutans in both therapeutic and preventive settings, albeit not fully. In contrast, Q10 completely preserves the viability of infected Her HEp-2 cells at all concentrations. The effects of the coexistence of Q10 and probiotics were not quite equal, among which L. salivarius and 5 µg of Q10 form the best results. The microscopic adherence assay of S. mutans revealed that samples containing Q10 had significantly lower adhesion of probiotics and S. mutans to HEp-2 cells. Similarly, plates containing L. salivarius with 5µg or L. plantarum with 1µg Q10 or sole presence of L. salivarius had the lowest S. mutans adherence among others. Also, L. salivarius with 5µg Q10 had one of the highest probiotic adherences. Conclusion: In conclusion, co-administration of Q10 and probiotics especially in presence of L. salivarius with 5µg Q10 could have remarkable effects on HEp-2 cell viability, S. mutans, and probiotic adherence. Nevertheless, our study, for the first time, showed that Q10 might have an anti-bacterial activity by suppressing the adhesion of tested bacteria to HEp-2 cells. This hypothesis, if correct, suggests that due to their different mechanisms, co-prescription of Q10 and probiotics may lead to better clinical responses, especially in the mentiond dose.

An antibacterial and self-healing hydrogel from aldehyde-carrageenan for wound healing applications.

This study aimed to develop and characterize a novel antibacterial, self-healing hydrogel made from aldehyde-carrageenan. Thus, carrageenan (CA) was first oxidized using different amounts of sodium periodate (NaIO4), and the highest concentration of aldehyde was obtained when the ratio of NaIO4 to CA was 1.5:1. Using dopamine (PDA) and zinc ions (Zn2+), various hydrogels were synthesized from oxidized carrageenan (O-CA). The effects of dopamine and zinc ions on the properties of O-CA hydrogel were examined. According to Fourier Transform Infrared Spectroscopy (FTIR) studies, the hydrogel's components are linked by Schiff bases, hydrogen bonds, and ion complexes. The rheological tests confirmed that hydrogels were elastic gels, not viscous sol, and were able to recover rapidly. Adding zinc to the hydrogel reduced weight loss (38 %) and provided extra antibacterial properties, particularly against E. coli. In addition, collagen secretion and cell attachment to Zn-containing hydrogels were significantly increased, and fibroblast viability reached 118 %. Overall, a hybrid O-CA/PDA/Zn hydrogel has excellent potential for wound healing applications.

Novel targets in rectal cancer by considering lncRNA-miRNA-mRNA network in response to Lactobacillus acidophilus consumption: a randomized clinical trial.

We aimed to explore the lncRNA-miR-mRNA network in response to Lactobacillus acidophilus (L. acidophilus) consumption in rectal cancer patients. The candidate miRs were first taken from the GEO and TCGA databases. We constructed the lncRNA-miR-mRNA network using the high-throughput sequencing data. At last, we created a heatmap based on the experimental data to show the possible correlation of the selected targets. The expression levels of selected targets were measured in the samples of 107 rectal cancer patients undergoing placebo and probiotic consumption and 10 noncancerous subjects using Real-Time PCR. Our analysis revealed a group of differentially expressed 12 miRs and 11 lncRNAs, and 12 genes in rectal cancer patients. A significant expression increase of the selected tumor suppressor miRs, lncRNAs, and genes and a substantial expression decrease of the selected oncomiRs, onco-lncRNAs, and oncogenes were obtained after the probiotic consumption compared to the placebo group. There is a strong correlation between some network components, including miR-133b and IGF1 gene, miR-548ac and MSH2 gene, and miR-21 and SMAD4 gene. In rectal cancer patients, L. acidophilus consumption was associated with improved expression of the lncRNA-miR-mRNA network, which may provide novel monitoring and therapeutic approaches.

Comparison of the effects of Lactobacillus plantarum versus imipenem on infected burn wound healing.

Background: Infection of burn wounds is one of the most important problems in the world. Lactobacillus plantarum is known for burn wound healing because of the immunomodulatory and anti-microbial roles. This study was performed to compare the effects of L. plantarum and imipenem - alone and in combination - on infected burn wound healing. Methods: Burn wounds were experimentally induced on 50 rats in three test groups (germ and supernatant of L. plantarum ) and two control groups (n=10 each) and were inoculated with Pseudomonas aeruginosa. During a 14-day period, wounds in all groups were daily treated topically. The data were analyzed using one-way analysis of variance followed by Tukey-Kramer and LSD. A p-value of < 0.05 was considered as statistically significant. Results: The mean size of the wound on day 14 after the treatment in the probiotic group was significantly lower than the control and the supernatant treated groups (p<0.05). The percentage of wound healing was significantly higher in the probiotic pellet treated group compared to the imipenem and the supernatant groups (by Anova test: 69.58%, p=0.022). The mean leukocyte count in the probiotic pellet group (12110) and supernatant group (13650) was significantly higher than the imipenem group (7670) (p=0.002 and 0.001, respectively). Wound cultures revealed that the percentage of cases where the pathogens had no growth was significantly different among the comparison groups. In all three test groups, P. aeruginosa was completely eliminated in comparison to the positive control group (p<0.05). Conclusion: The results of our study showed that L. plantarum and its by-products promote wound healing and can be used as an alternative to antibiotics to treat ulcer infections caused by resistant bacteria.

Effects of probiotic, cinnamon, and synbiotic supplementation on glycemic control and antioxidant status in people with type 2 diabetes; a randomized, double-blind, placebo-controlled study.

PURPOSE: The aim of this study was to investigate the effect of probiotic bacteria of Lactobacillus acidophilus, cinnamon powder and their combinations on the glycemic and antioxidant indices in patients with type 2 diabetes. METHODS: A total of 136 patients randomized with type 2 diabetes entered the study and were randomly divided into four groups who were matched for age and gender. Thereafter, alongside their routine pharmacotherapy, each group followed one of the following diets: Group A: Lactobacillus acidophilus 108 cfu and 0.5 g of powdered cinnamon (synbiotic). Group B: Lactobacillus acidophilus (probiotic), Group C: powdered cinnamon. Group D: rice flour powder as placebo. At the beginning and end of the intervention, fasting blood sugar (FBS), HbA1c, advance glycation end products (AGE), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and antioxidant enzymes of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) were measured. RESULTS: Following 3 months of treatment, the mean FBS level was decreased significantly in probiotic, cinnamon, and synbiotic supplementation groups compared with control (P < 0.01). FBS levels in probiotic, cinnamon, and synbiotic groups were significantly decreased compared with the control group (P = 0.001, P = 0.063 and P = 0.001, respectively). The mean HbA1C in probiotic, cinnamon, and synbiotic groups were also decreased (P = 0.001, P = 0.001 and P = 0.04, respectively). The mean AGE in synbiotic group was significantly decreased (P = 0.037). Probiotic, cinnamon and synbiotic all could improve antioxidant enzyme activity modestly. However, the most significant effect was seen in probiotic group. CONCLUSIONS: According to the current results, the use of probiotic supplements (individually or in combination with cinnamon) leads to a reduction in blood glucose and an increase in antioxidant enzymes in people with type 2 diabetes.

Therapeutic and Preventive Effects of Olea europaea Extract on Indomethacin-Induced Small Intestinal Injury Model in Rats.

BACKGROUND: Olea europaea (known as olive fruit) has anti-inflammatory and antioxidant activities and many potential health benefits including gastric inflammation reduction has been shown previously. This study aimed to investigate the preventive and therapeutic effects of O. europaea extract on the early histological changes in indomethacin-induced small intestinal injury model with the plasma D-lactate concentration being measured as a tool for determination of intestinal permeability. METHODS: In this experimental study, two separate protective and therapeutic protocols were designed. In both experiments, male Wistar rats were randomly divided into 4 groups and either pretreated with 0, 100, 200, or 400 mg/kg/day of O. europaea extract or received the treatment after administration of indomethacin. Their small intestines were examined to compare the histological changes. The intestinal injury severity was evaluated according to the presence of eosinophils, plasma cell infiltration, edema, congestion, and hyperplasia of the crypt using a histological scoring system. Also, measured were the presence of neutrophils, decreased villus length-to-crypt depth ratio, and destructed villus architecture. The plasma concentration of D-lactate was measured as well. RESULTS: The therapeutic use of O. europaea decreased the eosinophil, edema, congestion, and crypt hyperplasia scores compared to the control group. Although no significant difference was seen between groups of the preventive experiment in plasma cell infiltration score, villus length-to-crypt depth ratio, neutrophil infiltration, and percentage of destructed villus architecture, treatment with O. europaea caused a reduction in edema, eosinophil, congestion, and crypt hyperplasia score. In both experiments, no significant difference was seen between groups for villus length-to-crypt depth ratio, neutrophil infiltration, and percentage of destructed villus architecture. Plasma D-lactate concentration was decreased in all O. europaea-treated groups compared to the control group in the therapeutic and preventive experiments (p < 0.01, one-way ANOVA followed by the Dunnett test). CONCLUSION: O. europaea extract can be used to decrease some side effects of indomethacin on intestinal tissue and enhances the gastrointestinal function. O. europaea extract could be considered as a potential herbal supplement in the treatment of intestinal morphological injuries.

Altered levels of salivary biochemical markers in periodontitis.

PURPOSE: To investigate the association between periodontitis and levels of biochemical markers as well as enzyme activity. METHODS: Unstimulated whole saliva samples were obtained from 30 patients with periodontitis. Biochemical factors including the levels of malondialdehyde (MDA), superoxide dismutase (SOD), nitric oxide (NO), uric acid (UA), and lactoferrin, as well as ß-hexosaminidase (ß-HEX) activity were measured. RESULTS: The levels of a salivary oxidant such as MDA and NO were statistically significantly higher in periodontitis patients than to that of healthy individuals. Similarly, the results indicated elevated levels of lactoferrin and ß-HEX activity in saliva of the periodontitis group, which was statistically significant when compared to the controls. While the levels of an enzymatic antioxidant such as SOD were higher in the periodontitis patients than in the control subjects, uric acid levels were statistically significantly lower in the saliva of the periodontitis patients than in the healthy controls. CLINICAL SIGNIFICANCE: Except for uric acid, as a non-enzymatic antioxidant, the levels of salivary oxidative stress generally increase in the saliva of periodontitis patients. Since altered levels of salivary biomarkers such as oxidative stress and antioxidant substances might contribute in systemic and local complications in the patients, these informative biomarkers can be used as a promising factor for the early diagnosis of the disease.

The implication of single-nucleotide polymorphisms in ovarian hyperstimulation syndrome.

Ovarian hyperstimulation syndrome (OHSS) is an undesirable complication in the course of ovarian stimulation. This kind of stimulation is aimed at acquiring a sufficient number of high-quality oocytes in in vitro fertilization (IVF). Whereas the predisposition to OHSS could be impacted by genetic polymorphisms in susceptible genes, the present study has been jointly conducted with an Iranian cohort to scrutinize its relevant implication. Genomic DNA was extracted from blood samples of patients with a normal ovarian response (NOR) or with OHSS. Samples were analyzed to detect polymorphisms MTHFR rs1801131, MTHFR rs1801133, AMHR2 rs2002555, LHCGR rs2293275, PGR rs10895068, and SERPINE1 rs1799889. Variations of MTHFR, AMHR2, LHCGR, and PGR genes were significantly associated with the developing OHSS. After correction for multiple analysis, this difference was not evident for PGR genotypes. The polymorphic alleles of MTHFR (rs1801131 C-allele and rs1801133 T-allele), AMHR2 (rs2002555 G-allele), and LHCGR (rs2293275 G-allele) were significantly more prevalent among patients with OHSS compared to those in the NOR group. In contrast, the minor allele of PGR single-nucleotide polymorphism (SNP) (rs10895068, A-allele) was more prominent among patients with a NOR than those with OHSS. No significant difference was observed in genotypes or alleles of SERPINE1 rs1799889. The observations indicated that the minor alleles of MTHFR, AMHR2, and LHCGR genes could be considered an independent risk factor in susceptibility to OHSS. Nevertheless, polymorphic allele in the PGR rs10895068 SNP contributes to preventing OHSS occurrence. Therefore, it can be argued that these genes have a significant impact on OHSS.

Accelerated wound healing process in rat by probiotic Lactobacillus reuteri derived ointment.

Background Antioxidant agents have positive effects on wound healing process and on the other hand probiotics also have great antioxidation or oxidation-resistance properties. Methods In this study, we investigated the effects of Lactobacillus reuteri derived ointment as a probiotic bacterium on the wound healing process using a full-thickness rat skin model. The wound diameter and contraction percent measurement, histopathological examination, and myeloperoxidase (MPO) enzyme activity evaluation were carried out during the period of study. Results Amount of deposit collagen was significantly increased; epithelization speed and epidermis production were also observed in the ointment-treated group, while the inflammation rate was also significantly lower than the control group. In addition, a significant reduction in the activity of enzyme MPO was also detected in the ointment-treated group. Conclusions According to the results, probiotic ointment effectively accelerates the wound healing process and it can be used as a promising drug for wound dressing application.

In vitro osteogenic differentiation potential of the human induced pluripotent stem cells augments when grown on Graphene oxide-modified nanofibers.

Due to the several limitations that surgeons are faced during bone tissue implantation there are daily increases in introducing new cell-co-polymer composites for use in bone tissue engineering approaches. In this study tried to develop a suitable nanostructured bio-composite for enhancing osteogenic differentiation of the human induced pluripotent stem cells (iPSCs). Polyvinylidene fluoride-Graphene oxide (PVDF-GO) nanofibers was fabricated by electrospinning and then characterized using scanning electron microscope, tensile and viability assays. After that osteogenic differentiation of the iPSCs was investigated in three groups, including PVDF, PVDF-GO and tissue culture plate as a control group. Alkaline phosphatase activity and calcium content of the iPSCs cultured on PVDF-GO were significantly higher than those cultured on other groups. In addition, Runx2, osteocalcin and osteonectin genes were up regulated in iPSCs cultured on PVDF-GO significantly higher than those cells cultured on PVDF and control. Finally, osteocalcin and osteopontin proteins expression evaluated and the results confirmed higher osteoinductivity of the PVDF-GO nanofibers in comparison with the PVDF nanofibers. According to the results, it was demonstrated that PVDF-GO nanofibers have a great osteoinductive potential and taking together iPSCs-PVDF-GO nanofibrous construct can be an appropriate bio-implant to use for bone tissue engineering applications.

A comprehensive overview on utilizing electromagnetic fields in bone regenerative medicine.

Stem cells are one of the most important sources to develope  a new strategy for repairing bone lesions through tissue engineering. Osteogenic differentiation of stem cells can be affected by various factors such as biological, chemical, physiological, and physical ones. The application of ELF-EMFs has been the subject of many research in bone tissue engineering and evidence suggests that this exogenous physical stimulus can promote osteogenic differentiation in several types of  cells. The purpose of this paper is to review the current knowledge on the effects of EMFs on stem cells in bone tissue engineering studies. We recapitulated and analyzed 39 articles that were focused on the application of EMFs for bone tissue engineering purposes. We tabulated scattered information from these articles for easy use and tried to provide an overview of conducted research and identify the knowledge gaps in the field.

Probiotic Bacteria and their Supernatants Protect Enterocyte Cell Lines from Enteroinvasive Escherichia coli (EIEC) Invasion.

Probiotic microorganisms have attracted a growing interest for prevention and therapy of gastrointestinal disorders. Many probiotic strains have been shown to inhibit growth and metabolic activity of enteropathogenic bacteria as well as their adhesion and invasion to intestinal cells. In the present study, we evaluated the interference of bacteria-free supernatants (BFS) of cultures belonging to sixteen strains of lactobacilli and bifidobacteria, with invasion of enteroinvasive Escherichia coli (EIEC) strain, using human colonic adenocarcinoma cell lines, T84 and Caco2 cells. To assess invasion of Caco-2 and T84 cells by EIEC, and measure the number of pathogens inside the enterocytes, the gentamicin protection assay was conducted. In addition, three different invasion inhibition assays were designed; namelyco-incubation, pre-incubation and treatment with the BFS of probiotics. Data obtained and theoretical calculation showed that the most effective assay in the prevention of pathogen invasion was treatment withBFS.Besides, co-incubation assay was more valid than pre-incubation assay in invasion prevention.The obtained results suggest that probiotics may produce some metabolites that strongly prevent invasion of enteroinvasive E.coli into the small and large intestine. Also, probiotics are able to compete with or exclude pathogen invasion.

Probiotics Lactobacillus plantarum and bifidobacterium B94: cognitive function in demyelinated model.

BACKGROUND: Multiple Sclerosis (MS) is a disease of the immune system that creates damage of Learning and memory in that. Using probiotic supplements is recommended for preventing MS disease and improving memory. This study aimed to investigate the effect of Lactobacillus plantarum (LP) and bifidobacterium B94 (BB94), on acquisition phase of spatial memory in the local demyelination of rats` hippocampus. METHODS: In this study, 32 male Wistar rats were divided into control, damage group and treatment groups. Treatment groups were including (LP) and (BB94). After the induction of demyelination by 3 µl of EB into the right dentate gyrus of the hippocampus in treatment groups, 1.5×10(8) probiotic bacteria were administered by gavage for 28 days. Data was analyzed using one-way ANOVA and Tukey post-hoc tests (p≤0.05). RESULTS: Findings demonstrated that injection of EB caused a significant increase in traveled distance (p<0.01) and also escape latency (p<0.05) compared with control group. Also, effect administrations of (LP) and (BB94) on traveled distance and escape latency were reviewed, and it was determined that administration of them do not cause significant reduction in the traveled distance compared with the lesion group. Also mentioned probiotics has no significant effect on swimming speed compared with lesion and saline groups. CONCLUSION: According to some studies, probiotics have a positive impact on improving the performance of spatial memory and learning, although the results of the current study could not indicate finality of this assumption. It seems that more researches is needed on this subject.

Focal Injection of Ethidium Bromide as a Simple Model to Study Cognitive Deficit and Its Improvement.

INTRODUCTION: Memory and cognitive impairments are some of devastating outcomes of Multiple Sclerosis (MS) plaques in hippocampus, the gray matter part of the brain. The present study aimed to evaluate the intrahippocampal injection of Ethidium Bromide (EB) as a simple and focal model to assess cognition and gray matter demyelination. METHODS: Thirty Wistar rats were divided into three groups: control group, which received saline, as solvent of EB, into the hippocampus; and two experimental groups, which received 3 µL of EB into the hippocampus, and then, were evaluated 7 and 28 days after EB injection (n=10 in each group), using a 5-day protocol of Morris Water Maze (MWM) task as well as Transmission Electron Microscopy (TEM) assay. RESULTS: Seven days after EB injection, the behavioral study revealed a significance increase in travelled distance for platform finding in the experimental group compared to the control group. In addition, the nucleus of oligodendrocyte showed the typical clumped chromatin, probably attributed to apoptosis, and the myelin sheaths of some axons were unwrapped and disintegrated. Twenty-eight days after EB injection, the traveled distance and the time spent in target quadrant significantly decreased and increased, respectively in experimental groups compared to the control group. Also, TEM micrographs revealed a thin layer of remyelination around the axons in 28 days lesion group. DISCUSSION: While intracerebral or intraventricular injection of EB is disseminated in different parts of the brain and can affect the other motor and sensory systems, this model is confined locally and facilitates behavioral study. Also, this project could show improvement of memory function subsequent to the physiological repair of the gray matter of the hippocampus.

Tissue remodeling investigation in varicose veins.

Although the etiology of varicose veins remains unknown, recent studies have focused on endothelial cell integrity and function because the endothelium regulates vessel tone and synthesizes many pro- and anti-inflammatory factors. The aim of this study was to investigate the evidence involving the endothelium in the development of varicose vein disease. In addition, tissue remodeling was investigated in varicose veins to determine the expression of different types of collagen. Tissue specimens of superficial varicose veins and control saphenous vein were used for immunohistochemical and transmission electron microscope (TEM). α-smooth muscle actin, and collagen I, III, IV antibodies were applied for immunohistochemical investigation. Findings of this study showed alterations of the intima, such as focal intimal discontinuity and denudation of endothelium; and the media, such as irregular arrangements of smooth muscle cells and collagen fibres in varicose veins. Our findings showed some changes in terms of distribution of types I, III and IV collagen in the intima and media of varicose vein walls compared with controls. These alterations to the media suggest that the pathological abnormality in varicose veins may be due to the loss of muscle tone as a result of the breakup of its regular structure by the collagen fibres. These findings only described some changes in terms of distribution of these types of collagen in the intima and media of varicose vein walls which may result in venous wall dysfunction in varicosis.

Association of Helicobacter pylori infection with acute myocardial infarction.

OBJECTIVES: Helicobacter pylori infection has been linked to cardiovascular diseases (CVD) and several studies have reported its positive association with inflammatory response after acute myocardial infarction (AMI). On account of the importance of the inflammatory process in the development of CVD, we decided to examine the seroprevalence of H. pylori, the prevalence of CVD risk in the more virulent strains bearing the cytotoxin-associated protein (CagA), and the changes in C-reactive protein (CRP) as an inflammatory marker in Iranian patients with AMI. METHODS: A case-control study was designed to determine the seropositivity status of H. pylori and CagA in blood samples obtained from 500 patients with AMI and 500 control individuals without any evidence of clinical CVD. Serum and peripheral blood mononuclear cells were analyzed using the enzyme-linked immunosorbent assay and western blotting methods, respectively. CRP levels were also measured in all individuals. RESULTS: The prevalence of H. pylori infection and CagA status were significantly higher among the patients with AMI than the controls (66 vs. 20% and 75.7 vs. 30%, respectively); the odds ratio was 2.57 (95% confidence interval 1.89-3.49). CRP levels were significantly different in the patients compared with the controls (5.02±1.04 mg/l vs. 2.41±0.9 mg/l, respectively). CONCLUSION: Our results confirmed that the patients with AMI had a significantly higher prevalence of H. pylori infection and CagA seropositivity than the control population. Infection with H. pylori may influence AMI, which in our findings shows an association between H. pylori seropositivity and AMI through an inflammatory process.