Systems biology-derived genetic signatures of mastitis in dairy cattle : a new avenue for drug repurposing

Mastitis, a disease with high incidence worldwide, is the most prevalent and costly disease in the dairy industry. Gram-negative bacteria such as Escherichia coli (E. coli) are assumed to be among the leading agents causing acute severe infection with clinical signs. E. Coli, environmental mastitis pathogens, are the primary etiological agents of bovine mastitis in well-managed dairy farms. Response to E. Coli infection has a complex pattern affected by genetic and environmental parameters. On the other hand, the efficacy of antibiotics and/or anti-inflammatory treatment in E. coli mastitis is still a topic of scientific debate, and studies on the treatment of clinical cases show conflicting results. Unraveling the bio-signature of mastitis in dairy cattle can open new avenues for drug repurposing. In the current research, a novel, semi-supervised heterogeneous label propagation algorithm named Heter-LP, which applies both local and global network features for data integration, was used to potentially identify novel therapeutic avenues for the treatment of E. coli mastitis. Online data repositories relevant to known diseases, drugs, and gene targets, along with other specialized biological information for E. coli mastitis, including critical genes with robust bio-signatures, drugs, and related disorders, were used as input data for analysis with the Heter-LP algorithm. Our research identified novel drugs such as Glibenclamide, Ipratropium, Salbutamol, and Carbidopa as possible therapeutics that could be used against E. coli mastitis. Predicted relationships can be used by pharmaceutical scientists or veterinarians to find commercially efficacious medicines or a combination of two or more active compounds to treat this infectious disease

Prediction of key regulators and downstream targets of E. coli induced mastitis

Mastitis, an inflammatory response of mammary glands to invading bacteria, is one of the most economically costly diseases affecting dairy animals. E. coli can introduced as a major etiological agent of bovine mastitis in wellmanaged dairy farms. It is of great significance to understand the regulatory mechanisms by which the disease can be controlled. High-throughput technologies combined with novel computational systems biology tools have provided new opportunities for better understanding the molecular mechanisms that underlie disease. In the current study, the results of microarray meta-analysis research were used to perform a network analysis to potentially identify molecular mechanisms that regulate gene expression profile in response to E. coli mastitis. In our result, transcription factors, TP53, SP1, ligands, INS, IFNG, EGF, and protein kinases, MAPK1, MAPK14, AKT1, were identified as the key upstream regulators whereas protein kinases, MAPK3, MAPK8, MAPK14, ligands, VEGFA, IL10, an extracellular protein, MMP2, and a mitochondrial membrane protein, BCL2, were identified as the key downstream targets of differentially expressed genes. The results of this research revealed important genes that have the key functions in immune response, inflammation or mastitis which can provide the basis for strategies to improve the diagnosis and treatment of mastitis in dairy cows

Integration of machine learning and metaanalysis identifies the transcriptomic biosignature of mastitis disease in cattle

This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.Gram-negative bacteria such as Escherichia coli (E. coli) are assumed to be among the main agents that cause severe mastitis disease with clinical signs in dairy cattle. Rapid detection of this disease is so important in order to prevent transmission to other cows and helps to reduce inappropriate use of antibiotics. With the rapid progress in high-throughput technologies, and accumulation of various kinds of ‘-omics’ data in public repositories, there is an opportunity to retrieve, integrate, and reanalyze these resources to improve the diagnosis and treatment of different diseases and to provide mechanistic insights into host resistance in an efficient way. Meta-analysis is a relatively inexpensive option with good potential to increase the statistical power and generalizability of single-study analysis. In the current meta-analysis research, six microarray-based studies that investigate the transcriptome profile of mammary gland tissue after induced mastitis by E. coli infection were used. This meta-analysis not only reinforced the findings in individual studies, but also several novel terms including responses to hypoxia, response to drug, anti-apoptosis and positive regulation of transcription from RNA polymerase II promoter enriched by up-regulated genes. Finally, in order to identify the small sets of genes that are sufficiently informative in E. coli mastitis, the differentially expressed gene introduced by meta-analysis were prioritized by using ten different attribute weighting algorithms. Twelve meta-genes were detected by the majority of attribute weighting algorithms (with weight above 0.7) as most informative genes including CXCL8 (IL8), NFKBIZ, HP, ZC3H12A, PDE4B, CASP4, CXCL2, CCL20, GRO1(CXCL1), CFB, S100A9, and S100A8. Interestingly, the results have been demonstrated that all of these genes are the key genes in the immune response, inflammation or mastitis. The Decision tree models efficiently discovered the best combination of the meta-genes as bio-signature and confirmed that some of the top-ranked genes -ZC3H12A, CXCL2, GRO, CFB- as biomarkers for E. coli mastitis (with the accuracy 83% in average). This research properly indicated that by combination of two novel data mining tools, meta-analysis and machine learning, increased power to detect most informative genes that can help to improve the diagnosis and treatment strategies for E. coli associated with mastitis in cattle

Effect of soluble Carcinoembryonic antigen on myogenic and enterocytic differentiation

Introduction: Carcinoembryonic antigen (CEA), a glycophosphatidyl inositol (GPI) anchored glycoprotein, is over-expressed in various cancers, including colorectal carcinomas. Inhibition of cell differentiation can cause cancer. Previous studies show transfection and over-expression of CEA gene in myoblasts or colonocytes leads to inhibition of cell differentiation. We investigated whether soluble CEA has a role in inhibition of cell differentiation.Materials and Methods: Monolayer cultures of L6, C2C12, Caco-2 and CHO cell lines were grown in dulbecco's modification of eagle's medium (DMEM) containing 10% fetal bovine serum (FBS). For differentiation induction C2C12 and L6 cell lines media, was changed to DMEM containing 2% horse serum. CEA concentration was measured in harvested media by ELISA assay. H&E staining, CK assay and RT-PCR for myogenin gene were used for C2C12 differentiation experiments. Results: The results showed although commercial pure CEA and LS-180 conditioned media (contain high level of CEA) have inhibitory effects on Caco-2 differentiation, but it can also inhibit differentiation of control groups. We observed the effect of high concentration of CEA (5μg/ml) on L6 differentiation is identical with the effect of BSA as control. C2C12 differentiation was inhibited in response to LS-180 conditioned media at morphological level and myogenin expression. CK activity was significantly lower in LS-180 conditioned media (P= 0.0012) and CHO (P= 0.0002) conditioned media groups in comparison to control group.Conclusion: Our finding shows soluble CEA antigen did not have a significant effect on cell differentiation

Integrated co-expression analysis of regulatory elements (miRNA, lncRNA, and TFs) in bovine monocytes induced by Str. uberis

Abstract Non-coding RNAs, including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), together with transcription factors, are critical pre-, co-, and post-transcriptional regulators. In addition to their criteria as ideal biomarkers, they have great potential in disease prognosis, diagnosis, and treatment of complex diseases. Investigation of regulatory mechanisms in the context of bovine mastitis, as most common and economic disease in the dairy industry, to identify elements influencing the expression of candidate genes as key regulators of the mammary immune response is not yet fully understood. Transcriptome profiles (50 RNA-Seq and 50 miRNA-Seq samples) of bovine monocytes induced by Str. uberis were used for co-expression module detection and preservation analysis using the weighted gene co-expression network analysis (WGCNA) approach. Assigned mi-, lnc-, and m-modules used to construct the integrated regulatory networks and miRNA-lncRNA-mRNA regulatory sub-networks. Remarkably, we have identified 18 miRNAs, five lncRNAs, and seven TFs as key regulators of str. uberis-induced mastitis. Most of the genes introduced here, mainly involved in immune response, inflammation, and apoptosis, were new to mastitis. These findings may help to further elucidate the underlying mechanisms of bovine mastitis, and the discovered genes may serve as signatures for early diagnosis and treatment of the disease

Beneficial Effects of American Ginseng on Epididymal Sperm Analyses in Cyclophosphamide Treated Rats

Objective: This study aims to evaluate the protective effects of American ginseng administered by gastric intubation on sperm vital quality in adult male rats treated with cyclophosphamide (CP).Materials and Methods: In this experimental study, 28 Adult male Wistar rats were assigned to four groups, seven rats in each. The animals allocated to control, CP treated, Ginseng treated and CP-Ginseng treated groups. Rats were treated with CP (6.1 mg/kg/day, i.p) for 6 weeks. American ginseng was used at a dose of 500 mg/kg/day during treatment. Sperm analysis (motion, count, morphology and viability) were evaluated at the end of the experiments. Sperm motion was assessed by Computer-Assisted Sperm Analysis (CASA). The data were analyzed using GB stat software. Probability values of p<0.05 and p<0.01 were considered significant.Results: The epididymal sperm counts in the groups that received CP showed significant decreases compared to the control group. Also dead and abnormal sperms significantly increased following CP treatment compared with control. The motility of caudal sperm was reduced significantly with CP treatment. Therefore, according to the results of this study, co-administration of CP and American ginseng can improve these parameters.Conclusion: American ginseng can prevent the cytotoxic effects of CP on sperm quality factors

Combined effect of eight weeks high intensity resistance training with ginger supplementation on waist to hip ratio, body composition and body mass in obese women

Introduction: This study aimed to investigate the effects of eight weeks intensive resistance training combined with the ginger supplement on body composition, waist to hip ratio (WHR) and boday mass index (BMI) in obese women. Materials and Methods: Participants were 18 women (with an average age of 30.11 &plusmn; 06.45 years, weight 82.46 &plusmn; 12.40 kg, BMI 31.60 &plusmn; 03.99 kg per square meter). The participants were split into resistance training + daily Ginger supplements (n=10) and resistance training + placebo (n=8) group randomly. All groups performed high intensity resistance training for eight weeks, three sessions per week (totally 24 sessions). Resistance training per session includes 8 exercise with upper-lower orders and 80-85 percent of 1RM, three set with 20 s rest intervals. Before and after the eight-week training, the examiner administered a protocol to assess body composition using Bioelectrical Impedance BOCAX1. The supplementation group consumed capsules containing ginger supplement (Zintoma Manufacturing Co. Goldaroo) 1000 mg per day in the morning and evening after meal and placebo group received 500 mg of starch in gelatin capsules similar to ginger supplementation in a double blind way. The participants used supplements twice a day (morning and afternoon) after their meals. Results: Comparison of changes in different variables showed that only reductions in weight and fat mass were significantly different between groups. The placebo group had more reduction in their weight and fat mass while ginger plus group showed less reduction. In both groups, other variables showed reductions too but the changes did not reach the significance level (0.05 Conclusion: It seems that resistance training alone can have more positive effect compared to resistance training plus ginger supplements capsule. Perhaps long-term use of ginger supplements or changes in its usage level combined with resistance training even repetition of the present study in a larger group may provide different results. Future studies can take a fresh look to resistance exercise and herbal supplements in different demographic groups to see their effects

Effects of Mineralocorticoid Receptors Blockade on FearMemory Reconsolidation in Rats

Reconsolidation memory is defined as a process in which the retrieval of a previously consolidated memory returns to a labile state which is then subject to stabilization. Previous studies have shown that mineralocorticoid receptors (MRs) modulate distinct phases of learning and memory, which display a high concentration and distinct distribution in the hippocampus. Moreover, we found no studies that examined the role of hippocampal MRs in fear memory reconsolidation. Here, we investigated the effect of MRs blockade on fear memory reconsolidation in rats. Additionally, to test whether blockade of protein synthesis would disrupt fear memory reconsolidation in our paradigm, we tested the effect of cycloheximide, an inhibitor of protein synthesis after memory reactivation. Results indicated that systemic as well as intra-hippocampal administrations of the MR antagonist spironolactone immediately following memory reactivation did not affect on post-retrieval long-term memory. Cycloheximide given after the reactivation treatment produced a strong impairment that persisted over test sessions. These findings indicate that MRs are not required for reconsolidation of fear-based memory