Ameliorating inflammation in an in vitro model by screening the anti-inflammatory and immunomodulatory roles of putative probiotics in inflammatory bowel disease.

IBD is considered a relapsing disease with relapsing phases. Probiotics are beneficial microorganisms that modulate inflammatory signaling pathways. Our aim was to identify the precise molecular effects of probiotics on inflammatory signaling pathways during the presence of inflammation. Evaluation of the expression of JAK/STAT and inflammatory genes after treatment of the HT -29 cell line with the sonicated pathogens and probiotics, simultaneously was performed by quantitative real-time polymerase chain reaction (qPCR) assay. The production of IL-6 and IL-1ß after administration of probiotics was conducted by means of cytokine assay. The probiotic cocktail resulted in the downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-кB pathway compared with Sonicat-treated cells. The expression of JAK/STAT genes was various after probiotic treatment. The application of probiotics has been observed to result in a notable decrease in the production of IL-6 and IL-1ß. The investigated probiotic cocktail, especially Bifidobacterium spp. showed anti-inflammatory effects on HT -29 cells via modulation of JAK/STAT and NF-кB signaling pathways. The use of probiotics with the least side effects could be considered a suitable treatment for patients with inflammatory bowel disease, even at the beginning of inflammation.

Investigation of the anti-inflammatory effects of native potential probiotics as supplementary therapeutic agents in an in-vitro model of inflammation.

BACKGROUND: IBD is considered an inflammatory disease with abnormal and exaggerated immune responses. To control the symptoms, different theraputic agents could be used, however, utilizing the agents with the least side effects could be important. Probiotics as beneficial microorganisms are one of the complementory theraputic agents that could be used to modulate inflammatory signaling pathways. In the current study, we aimed to identify the precise molecular effects of potential probiotics on signaling pathways involved in the development of inflammation. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK /STAT (JAK1, JAK2, JAK3, TYK2, STAT1, STAT2, STAT3, STAT4, STAT5 and STAT6) and inflammatory genes (NEMO, TIRAP, IRAK, and RIP) after the HT -29 cell line treatment with the sonicated pathogens and potential probiotics. A cytokine assay was also used to evaluate IL -6 and IL -1ß production after potential probiotic treatment. RESULTS: The potential probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared with cells that were treated with sonicated gram negative pathogens. The expression of STAT genes was different after potential probiotic treatment. The production of IL -6 and IL -1ß decreased after potential probiotic treatment. CONCLUSIONS: Considering the importance of controlling the symptoms of IBD to improve the life quality of the patients, using probiotic could be crucial. In the current study the studied native potential probiotic cocktails showed anti-inflammatory effects via modulation of JAK /STAT and NF-kB signaling pathways. This observation suggests that our native potential probiotics consumption could be useful in reducing intestinal inflammation.

Prophylactic vs. Therapeutic Effect of Probiotics on the Inflammation Mediated by the NF-κB Pathway in Inflammatory Bowel Conditions.

Probiotic supplements consumed adequately at the proper time can affect health by modulating inflammatory pathways in gastrointestinal epithelial cells and modifying the resultant inflammatory response. The current study applied in vitro models to investigate the effectiveness of probiotics in modulating inflammatory pathways and altering inflammatory gene expression in gastrointestinal epithelial cells, with the ultimate goal of promoting probiotic consumption as a therapeutic and preventive measure for chronic inflammatory bowel conditions. HT-29 cells were treated with Gram-negative bacteria to evaluate the changes in pathways related to inflammation activities before and after treatment with a Lactobacillus spp. cocktail (L. plantarum, L. rhamnosus, L. brevis, and L. ruteri) and a Bifidobacterium spp. cocktail (B. bifidum, B. langum, and B. breve) using the real-time PCR method and ELISA for IL-1ß and IL-6 as pro-inflammatory cytokines. The results showed that the expression of NF-κB signaling pathway genes and IL-1ß and IL-6 cytokines increased after exposure to Gram-negative components. In contrast, all probiotic combinations significantly decreased the expression of genes and the secretion of cytokines. However, this decrease was significantly smaller in cells that underwent probiotic treatment after inflammation induction. In addition, cocktails containing combined Lactobacillus and Bifidobacterium demonstrated robust anti-inflammatory activity relative to solo cocktails. Our observations confirm that probiotic consumption could positively impact inflammatory conditions and alleviate inflammatory symptoms; they can be particularly effective as a preventive measure. Our study provides preliminary evidence to support the lifetime consumption of probiotics.

The Role of Combining Probiotics in Preventing and Controlling Inflammation: A Focus on the Anti-Inflammatory and Immunomodulatory Effects of Probiotics in an In Vitro Model of IBD.

Objective: IBD is an inflammatory disease with abnormalities such as dysbiosis and abnormal immune system activity. Probiotics, as live beneficial microorganisms, play a role in maintaining health through various mechanisms, including the modulation of the immune system and the control of inflammation. Here, we aimed to investigate the efficacy of a probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. in modulating JAK/STAT and NF-kB inflammatory signaling pathways. Method: A quantitative real-time polymerase chain reaction (qPCR) assay was conducted to analyze the expression of JAK/STAT and inflammatory genes after treatment with the probiotic mixture before, after, and simultaneously with the sonicated pathogen in the HT-29 cell line. The production of IL-6 and IL-1ß after probiotic treatment was investigated via cytokine assay. Results: Treatment with probiotics resulted in downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway and JAK/STAT genes compared with sonicat-treated cells as inflammation inducers. The production of IL-6 and IL-1 decreased after probiotic treatment. Conclusions: The probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. showed anti-inflammatory effects by modulating JAK/STAT and NF-kB signaling pathways. The use of probiotics could be considered as an appropriate complementary treatment for patients with inflammatory bowel disease.

Decreased mucosal adhesion of Lactobacillus species in patients with inflammatory bowel disease.

Background: Probiotic Lactobacillus spp. modulate immune response via interactions of their binding proteins with epithelial cells. We studied the presence of attachment protein-encoding genes (mub1, mub2, and mapA) in Lactobacillus strains with probiotic features isolated from inflammatory bowel disease (IBD) patients and their attachment strength relative to healthy individuals. Methods: Bacterial strains have been isolated from stool samples of 35 healthy and 23 IBD volunteers. Lactobacillus spp. were identified using PCR. Strains with probiotic features were determined by testing resistance against acid and bile. Isolates were assigned as non-adhesive, adhesive, and strongly adhesive strains based on the number of attached bacteria to epithelial cells. Finally, PCR was used to detect the presence of mub1, mub2, and mapA genes. Results: Probiotic lactobacilli were isolated from 35/35 and 9/23 of healthy and IBD individuals and yielded a total of 87 and 28 strains, respectively. The Mub1 gene was detected in 95.4% and 100% (p>0.05), mub2 in 95.4% and 89.3% (p>0.05), and mapA in 94.3% and 78.6% (p<0.05) of healthy and IBD isolates, respectively. The numbers of bacteria attached to epithelial cells in healthy and IBD isolates were respectively 33.68±6.00 and 12.23±3.87 in non-adhesive, 71.3±10.83 and 42.17±1.33 in adhesive, 124.40±8.59 and 104.67±5.50 in the strongly adhesive group (p< 0.05). Conclusion: Less Lactobacillus spp. with weaker attachments to epithelial cells colonize the gut in IBD than healthy individuals. These findings suggest the beneficial role of probiotics in the management of IBD.

Anti-inflammatory and immunomodulatory effects of Lactobacillus spp. as a preservative and therapeutic agent for IBD control.

BACKGROUND: Probiotics have a beneficial effect on inflammatory responses and immune regulation, via Janus kinase/signal transduction and activator of transcription (JAK/STAT) and NF-κB signaling pathways. To evaluate the precise effects of Lactobacillus spp. as a protective and therapeutic agent, we aimed to investigate the efficacy of Lactobacillus spp. in modulating JAK/STAT and nuclear factor kappa B (NF-κB) inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIR-associated Protein [TIRAP], Interleukin 1 Receptor Associated Kinase[IRAK4], Nuclear factor-kappa B Essential Modulator [NEMO], and receptor interacting protein [RIP]) followed by treatment of the HT-29 cell line with sonicated pathogens before, after, and simultaneously with Lactobacillus spp. A cytokine assay was also used to evaluate interleukin (IL)-6 and IL-1ß production after treatment with Lactobacillus spp. RESULTS: Lactobacillus spp. downregulated JAK and TIRAP, IRAK4, NEMO, and RIP genes in the NF-κB pathway compared to sonicate-treated cells. The expression of STAT genes was different after treatment with probiotics. The production of IL-6 and IL-1ß decreased after probiotic treatment. CONCLUSIONS: Our Lactobacillus spp. cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-κB signaling pathways in all three treatment variants. Therefore, Lactobacillus spp. as a dietary supplement can both prevent and reduce inflammation-related diseases such as inflammatory bowel disease.

The effects of the probiotic cocktail on modulation of the NF-kB and JAK/STAT signaling pathways involved in the inflammatory response in bowel disease model.

BACKGROUND: Probiotics positively affect inflammatory responses, in part, through Janus kinase/signal transduction and activator of transcription (JAK/STAT) and inflammatory signaling pathways. To evaluate the precise effects of probiotics as protective treatment, we aimed to investigate the effectiveness of Lactobacillus spp., Bifidobacterium spp., and a mixture of these probiotics in modulating the JAK/STAT and inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIRAP, IRAK4, NEMO, and RIP) following HT-29 cell line treatment with sonicated pathogens Lactobacillus spp., Bifidobacterium spp., and a mixed cocktail. A cytokine assay was also used to evaluate the IL-6 and IL-1ß production following the probiotic treatment. RESULTS: The probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared to sonicate pathogen treatment cells. The expression of STAT genes was variable following probiotic treatment. The IL-6 and IL-1ß production decreased after probiotic treatment. CONCLUSIONS: Our probiotic cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-kB pathways. Therefore, Lactobacillus spp. and Bifidobacterium spp. probiotics as nutritional supplements may reduce inflammation-associated diseases such as inflammatory bowel disease (IBD).

Effects of green synthesis of sulfur nanoparticles from Cinnamomum zeylanicum barks on physiological and biochemical factors of Lettuce (Lactuca sativa).

In this study, the effect of green synthesized sulfur nanoparticle (SNP) at different concentration (0, 0.01, 0.1, 1 and 10 mg/ml) on some physiological, phytochemical and biochemical traits of lettuce plants was investigated. For the first time, SNP were green synthesized using Cinnamomum zeylanicum barks extract. Our results indicated that the treatment of lettuce plants with 1 mg/ml of SNP improved the growth and photosynthetic parameters of lettuce plants than related control. Some other physiological parameters such as proline, glycine betaine and soluble sugars levels along with some phytochemical parameters like anthocyanin, total phenol, flavonoids and tannin contents were enhanced after treatment of the plants with same concentration of SNP. On the other hand, specific activity of antioxidant enzymes (catalase, ascorbate peroxidase and polyphenol oxidase) and stress markers level, MDA and H2O2 were reduced in the same treated lettuce plants. However, a concentration of 10 mg/ml of SNP exhibited toxicity on lettuce plants with inducing oxidative stress markers (H2O2 and MDA) and consequently reducing plant growth and biomass. This oxidative stress tend to diminish some physiological, phytochemical and biochemical parameters in treated lettuce plants. Overall, it can be concluded that the green synthesis SNP at an optimal concentration of 1 mg/ml improved physiological parameters in the lettuce plants making them potent to tolerate stressful conditions. However, higher concentration of SNP (10 mg/ml) indicated toxic effects on all of the physiological parameters.

Piperlongumine as anticancer agent: The story so far about killing many birds with one stone.

Piperlongumine is a biologically and pharmacologically active constituent of the plant Piper longum. This compound is gradually gaining attention because of its ability to inhibit/prevent different cancers. Modern era of molecular oncology is incomplete without ground-breaking discoveries made in the field of cell signaling pathways. High-throughput technologies have considerably improved our understanding about wide ranging signal transduction cascades which play crucial role in cancer development and progression. It is exciting to note that piperlongumine has been shown to pleiotropically modulate different oncogenic signaling pathways. We partition this multi-component review into discrete sections and categorically summarize key findings related to excellent ability of piperlongumine to therapeutically target JAK-STAT, NF-kB and PI3K/AKT/mTOR pathways. We also set spotlight on regulation of TRAIL pathway and autophagy by piperlongumine in different cancers. On the basis of the current understanding of piperlongumine, molecular biologists and pharmacologists will develop the next generation of translational studies, which will prove to be helpful in improving the clinical outcome and getting a step closer to personalized medicine.

Regulatory network analysis of Epstein-Barr virus identifies functional modules and hub genes involved in infectious mononucleosis.

Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis (IM) and establishes lifetime infection associated with a variety of cancers and autoimmune diseases. The aim of this study was to develop an integrative gene regulatory network (GRN) approach and overlying gene expression data to identify the representative subnetworks for IM and EBV latent infection (LI). After identifying differentially expressed genes (DEGs) in both IM and LI gene expression profiles, functional annotations were applied using gene ontology (GO) and BiNGO tools, and construction of GRNs, topological analysis and identification of modules were carried out using several plugins of Cytoscape. In parallel, a human-EBV GRN was generated using the Hu-Vir database for further analyses. Our analysis revealed that the majority of DEGs in both IM and LI were involved in cell-cycle and DNA repair processes. However, these genes showed a significant negative correlation in the IM and LI states. Furthermore, cyclin-dependent kinase 2 (CDK2) - a hub gene with the highest centrality score - appeared to be the key player in cell cycle regulation in IM disease. The most significant functional modules in the IM and LI states were involved in the regulation of the cell cycle and apoptosis, respectively. Human-EBV network analysis revealed several direct targets of EBV proteins during IM disease. Our study provides an important first report on the response to IM/LI EBV infection in humans. An important aspect of our data was the upregulation of genes associated with cell cycle progression and proliferation.

Prevalence of Cytomegalovirus in Patients With Multiple Sclerosis: A Case-Control Study in Northern Iran.

BACKGROUND: Multiple sclerosis (MS) is a chronic debilitating disease known as one of the most common neurological dysfunctions in young adults. Recent studies suggest that infections with herpesviruses play a critical role in the pathogenesis of MS. OBJECTIVES: The present investigation aimed to detect the presence of cytomegalovirus (CMV) in patients with MS using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) methods. PATIENTS AND METHODS: Plasma and peripheral blood mononuclear cells (PBMCs) were collected from MS patients (n = 82) and from blood donors as control group (n = 89). They were tested for the presence of CMV antibodies and DNA by ELISA and PCR, respectively. RESULTS: Anti-CMV was positive in 65 (79.3%) and 69 (77.5%) of the MS patients and healthy subjects, respectively (P= 0.853). Similarly, 23 (28%) and 2 (2.2%) patients were positive for CMV DNA among the MS and control groups, respectively. Statistical analysis showed that the frequency of CMV DNA in the MS patients was significantly higher than in the healthy controls (P < 0.001). CONCLUSIONS: The results of this study showed a possible association between CMV infection and MS. Further experimental and epidemiological studies using case-control approaches are needed to confirm this association.

Impacts of the G145R Mutation on the Structure and Immunogenic Activity of the Hepatitis B Surface Antigen: A Computational Analysis.

BACKGROUND: Vaccine-escaped hepatitis B virus (HBV) mutations occur within the "a" determinant area, which is located in the major hydrophilic region (MHR) of the hepatitis B surface antigen (HBsAg) protein. It is now well established that the common G145R mutation is highly capable of escaping from HBsAg immune recognition. However, the impacts of this mutation on the structure and immunogenic activity of HBsAg have been poorly investigated. OBJECTIVES: The present study analyzed the effects of the G145R mutation on the structure and immunogenic activity of the HBsAg. MATERIALS AND METHODS: Three-dimensional (3D) structure of HBsAg for both the wild-type and G145R mutant were predicted and refined using several web tools. After quantitative evaluations, the effects of the G145R mutation on the secondary and 3D structures of the HBsAg were investigated. In parallel, the immunogenic activity of the wild-type and mutant HBsAg was also analyzed using a ClusPro docking server as well as the IEDB web tool. Further analyses were performed via molecular dynamics (MD) simulations using the GROMACS v5.0.2 simulation package. RESULTS: The G145R mutation causes a considerable reduction in the immunogenic activity of the HBsAg through a conformational change in the HBsAg antigenic loops. This mutation inserts a new ß-strand in the "a" determinant region of the HBsAg, leading to a reduced binding affinity to its monoclonal antibody, MAb12. The G145R mutation also increased the compactness and stability of the HBsAg by enhancing the rigidity of the "a" determinant. CONCLUSIONS: These data will be beneficial for designing more advanced antibodies for the recognition of the HBsAg in diagnostics. In addition, the results of this study may assist in the design or development of more effective hepatitis B vaccines.

How Hepatitis C Virus Leads to Hepatocellular Carcinoma: A Network-Based Study.

BACKGROUND: Hepatitis C virus (HCV) has been known as a major cause of hepatocellular carcinoma (HCC) worldwide. However, the distinct molecular mechanisms underlying the effects of HCV proteins on the HCC progression have remained unclear. OBJECTIVES: In the present study, we studied the possible role of HCV in the HCC initiation and invasion using topological analysis of protein-protein interaction (PPI) networks. MATERIALS AND METHODS: After analysis with GEO2R, a PPI network of differentially expressed genes (DEGs) was constructed for both chronic HCV and HCC samples. The STRING and GeneMANIA databases were used to determine the putative interactions between DEGs. In parallel, the functional annotation of DEGs was performed using g: Profiler web tool. The topological analysis and network visualization was carried outperformed using Cytoscape software and the top hub genes were identified. We determined the hub genes-related miRNAs using miRTarBase server and reconstructed a miRNA-Hubgene network. RESULTS: Based on the topological analysis of miRNA-Hubgene network, we identified the key hub miRNAs. In order to identify the most important common sub-network, we aligned two PPI networks using NETAL tool. The c-Jun gene was identified as the most important hub gene in both HCV and HCC networks. Furthermore, the hsa-miR-34a, hsa-miR-155, hsa-miR-24, hsa-miR-744 and hsa-miR-92a were recognized as the most important hub miRNAs with positive correlation in the chronic HCV and HCC samples. Functional annotation of differentially expressed miRNAs (DEMs) using the tool for annotations of human miRNAs (TAM) revealed that there is a considerable overlap between miRNA gene expression profiles of HCV-infected and HCC cells. CONCLUSIONS: Our results revealed the possible crucial genes and miRNAs involved in the initiation and progression of HCC cells infected with HCV.

The High Prevalence of the Varicella Zoster Virus in Patients With Relapsing-Remitting Multiple Sclerosis: A Case-Control Study in the North of Iran.

BACKGROUND: Multiple sclerosis (MS) is the most common neurological autoimmune disease, characterized by multifocal areas of inflammatory demyelination within the central nervous system. It has been hypothesized that the stimulation of the immune system by viral infections is the leading cause of MS among susceptible individuals. OBJECTIVES: The aim of this study was to investigate the prevalence of the varicella zoster virus (VZV) in patients with relapsing-remitting multiple sclerosis. PATIENTS AND METHODS: Plasma and peripheral blood mononuclear cells (PBMCs) collected from MS patients (n = 82) and controls (n = 89) were screened for the presence of anti-VZV antibodies and VZV DNA by the ELISA and PCR methods. DNA was extracted from all samples, and VZV infection was examined by the PCR technique. Statistical analysis was used to investigate the frequency of the virus in MS patients and a healthy control group. RESULTS: Of all the MS patients, 78 (95.1%) and 21 (25.6%) were positive for anti-VZV and VZV DNA, respectively. Statistical analysis of the PCR results showed a significant correlation between the abundance of VZV and MS disease (P < 0.001). However, there was no significant correlation between the abundance of anti-VZV antibodies and MS disease by the ELISA method. CONCLUSIONS: These results support the hypothesis that VZV may contribute to MS in establishing a systemic infection process and inducing an immune response.