Moringa leaf meal exerts growth benefits in small ruminants through modulating the gastrointestinal microbiome.

This study investigated the impact of feeding 17% moringa leaf meal (MLM) on the ruminal and fecal microbial composition and body weight gain (BWG) performance of lambs (Ovis aries) and kids (Capra hircus). A total of n = 28 lambs (n = 14, no-moringa, n = 14, 17% moringa) and 24 kids (n = 12, no-moringa, n = 12, 17% moringa) were involved in the experiment and body weight was recorded fortnightly. Metagenomic shotgun sequencing was performed on 28, 22, and 26 ruminal solid, liquid fraction, and fecal samples from lambs, and 23, 22, and 23 samples from kids. Moringa supplementation significantly increased BWG in lambs (21.09 ± 0.78 to 26.12 ± 0.81 kg) and kids (14.60 ± 1.29 to 18.28 ± 1.09 kg) (p-value ≤ 0.01). Microbiome analysis revealed an elevated Firmicutes:Bacteroidetes ratio in the moringa diet group. Moringa-fed animals exhibited increased microbial genera associated with volatile fatty acids (VFAs) production (Prevotella, Anaerovibrio, Lachnospiraceae, Butyrivibrio, Christensenella) and starch and fiber digesters (Proteobacteria, Ruminococcus). The increase in the bacterial genus Sharpea suggested possible methane reduction and decreased proportion of pathogens, Aliarcobacter_ID28198, Campylobacter_ID194 and Campylobacter_ID1660076 suggest health benefits. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated significant alterations in microbial gene pool and metabolic pathways related to carbohydrate, protein, lipid and energy metabolism, indicating potential improvements in animal health. Overall, moringa feeding showed higher energy recovery, improved growth, and potential benefits in methane reduction and reduced pathogenic bacteria.

Interplay of gene expression and regulators under salinity stress in gill of Labeo rohita.

BACKGROUND: Labeo rohita is the most preferred freshwater carp species in India. The concern of increasing salinity concentration in freshwater bodies due to climate change may greatly impact the aquatic environment. Gills are one of the important osmoregulatory organs and have direct contact with external environment. Hence, the current study is conducted to understand the gill transcriptomic response of L. rohita under hypersalinity environment. RESULTS: Comprehensive analysis of differentially expressed long non-coding RNAs (lncRNAs), microRNAs (miRNAs) and mRNAs was performed in gills of L. rohita treated with 2, 4, 6 and 8ppt salinity concentrations. Networks of lncRNA-miRNA-mRNA revealed involvement of 20, 33, 52 and 61 differentially expressed lncRNAs, 11, 13, 26 and 21 differentially expressed miRNAs in 2, 4, 6 and 8ppt groups between control and treatment respectively. These lncRNA-miRNA pairs were regulating 87, 214, 499 and 435 differentially expressed mRNAs (DE mRNAs) in 2, 4, 6 and 8ppt treatments respectively. Functional analysis of these genes showed enrichment in pathways related to ion transportation and osmolyte production to cope with induced osmotic pressure due to high salt concentration. Pathways related to signal transduction (MAPK, FOXO and phosphatidylinositol signaling), and environmental information processing were also upregulated under hypersalinity. Energy metabolism and innate immune response pathways also appear to be regulated. Protein turnover was high at 8ppt as evidenced by enrichment of the proteasome and aminoacyl tRNA synthesis pathways, along with other enriched KEGG terms such as apoptosis, cellular senescence and cell cycle. CONCLUSION: Altogether, the RNA-seq analysis provided valuable insights into competitive endogenous (lncRNA-miRNA-mRNA) regulatory network of L. rohita under salinity stress. L. rohita is adapting to the salinity stress by means of upregulating protein turnover, osmolyte production and removing the damaged cells using apoptotic pathway and regulating the cell growth and hence diverting the essential energy for coping with salinity stress.

Resistance Evolution against Host-directed Antiviral Agents: Buffalopox Virus Switches to Use p38-ϒ under Long-term Selective Pressure of an Inhibitor Targeting p38-α.

Host-dependency factors have increasingly been targeted to minimize antiviral drug resistance. In this study, we have demonstrated that inhibition of p38 mitogen-activated protein kinase (a cellular protein) suppresses buffalopox virus (BPXV) protein synthesis by targeting p38-MNK1-eIF4E signaling pathway. In order to provide insights into the evolution of drug resistance, we selected resistant mutants by long-term sequential passages (P; n = 60) in the presence of p38 inhibitor (SB239063). The P60-SB239063 virus exhibited significant resistance to SB239063 as compared to the P60-Control virus. To provide mechanistic insights on the acquisition of resistance by BPXV-P60-SB239063, we generated p38-α and p38-ϒ (isoforms of p38) knockout Vero cells by CRISPR/Cas9-mediated genome editing. It was demonstrated that unlike the wild type (WT) virus which is dependent on p38-α isoform, the resistant virus (BPXV-P60-SB239063) switches over to use p38-ϒ so as to efficiently replicate in the target cells. This is a rare evidence wherein a virus was shown to bypass the dependency on a critical cellular factor under selective pressure of a drug.

lncRNA-miRNA-mRNA network in kidney transcriptome of Labeo rohita under hypersaline environment.

The present study describes the kidney transcriptome of Labeo rohita, a freshwater fish, exposed to gradually increased salinity concentrations (2, 4, 6 and 8ppt). A total of 10.25 Gbps data was generated, and a suite of bioinformatics tools, including FEELnc, CPC2 and BLASTn were employed for identification of long non-coding RNAs (lncRNAs) and micro RNAs (miRNAs). Our analysis revealed a total of 170, 118, 99, and 269 differentially expressed lncRNA and 120, 118, 99, and 124 differentially expressed miRNAs in 2, 4, 6 and 8 ppt treatment groups respectively. Two competing endogenous RNA (ceRNA) networks were constructed i.e. A* ceRNA network with up-regulated lncRNAs and mRNAs, down-regulated miRNAs; and B* ceRNA network vice versa. 2ppt group had 131 and 83 lncRNA-miRNA-mRNA pairs in A* and B* networks, respectively. 4ppt group featured 163 pairs in A* network and 191 in B* network, while the 6ppt had 103 and 105 pairs. 8ppt group included 192 and 174 pairs. These networks illuminate the intricate RNA interactions in freshwater fish to varying salinity conditions.

Photocytotoxic kinetically stable ruthenium(II)-N,N-donor polypyridyl complexes of oxalate with anticancer activity against HepG2 liver cancer cells.

Liver cancer is one of the leading causes of death that motivating scientists worldwide to synthesize novel chemotherapeutics. Ru(II)-polypyridyl complexes are extensively studied for possible therapeutic and cellular applications due to their tunable coordination chemistry, structural diversity, ligand-exchange kinetics, accessible redox states, and rich photophysical or photochemical properties. Herein, we have synthesized a series of Ru(II) polypyridyl complexes [RuII(N^N)2(ox)] (1-3), where ox is oxalate (C2O42-) and N^N is 1,10-phenanthroline (phen) (1), dipyrido[3,2-d:2',3'-f]quinoxaline (dpq) (2), and dipyrido[3,2,-a:2',3'-c]phenazine (dppz) (3). Oxalate (ox2-) was opted as a bioactive dioxo ligand to prevent facile hydrolysis in aqueous media, thereby increasing the stability of the Ru(II)-polypyridyl complexes in physiological media. We thoroughly characterized all the complexes using ESI-MS, FT-IR, UV-vis, and 1H NMR spectroscopy and other physicochemical methods. The complexes were stable under physiological conditions and under low-energy green LED light (λirr = 530 nm). However, the photoirradiation of complexes resulted in the efficient generation of singlet oxygen (1O2) as a major reactive oxygen species (ROS). The role of the extended aromatic conjugation of the N^N-donor ligands in the complexes was demonstrated by their binding propensities with CT-DNA and bovine serum albumin (BSA). Both DNA intercalation and groove binding were evidenced, while tryptophan (Trp) and tyrosine (Tyr) binding site preferences were revealed from the synchronous fluorescence spectra (SFS) of BSA. The cytotoxic profiling of the complexes performed on hepatocellular carcinoma cells (HepG2) in the dark and in the presence of green light indicated their dose-dependent cytotoxicity. The [RuII(N^N)2(ox)] complexes exhibited enhanced photocytotoxicity mediated by efficient generation of cytotoxic 1O2 and effective interaction with DNA. All the complexes were internalized by the HepG2 liver cancer cells efficiently and localized to the cytoplasm and nucleus. The complexes exhibited potent anti-proliferative, anti-clonogenic, and anti-migratory effects on the cancer cells, suggesting their potential for therapeutic applications.

Resistant cumin cultivar, GC-4 counters Fusarium oxysporum f. sp. cumini infection through up-regulation of steroid biosynthesis, limonene and pinene degradation and butanoate metabolism pathways.

Cumin (Cuminum cyminum L.), an important spice crop belonging to the Apiaceae family is infected by Fusarium oxysporum f. sp. cumini (Foc) to cause wilt disease, one of the most devastating diseases of cumin adversely affects its production. As immune responses of cumin plants against the infection of Foc are not well studied, this research aimed to identify the genes and pathways involved in responses of cumin (cv. GC-2, GC-3, GC-4, and GC-5) to the wilt pathogen. Differential gene expression analysis revealed a total of 2048, 1576, 1987, and 1174 differentially expressed genes (DEGs) in GC-2, GC-3, GC-4, and GC-5, respectively. In the resistant cultivar GC-4 (resistant against Foc), several important transcripts were identified. These included receptors, transcription factors, reactive oxygen species (ROS) generating and scavenging enzymes, non-enzymatic compounds, calcium ion (Ca2+) transporters and receptors, R-proteins, and PR-proteins. The expression of these genes is believed to play crucial roles in conferring resistance against Foc. Gene ontology (GO) analysis of the up-regulated DEGs showed significant enrichment of 19, 91, 227, and 55 biological processes in GC-2, GC-3, GC-4, and GC-5, respectively. Notably, the resistant cultivar GC-4 exhibited enrichment in key GO terms such as 'secondary metabolic process', 'response to reactive oxygen species', 'phenylpropanoid metabolic process', and 'hormone-mediated signaling pathway'. Furthermore, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed the enrichment of 28, 57, 65, and 30 pathways in GC-2, GC-3, GC-4, and GC-5, respectively, focusing on the up-regulated DEGs. The cultivar GC-4 showed enrichment in pathways related to steroid biosynthesis, starch and sucrose metabolism, fatty acid biosynthesis, butanoate metabolism, limonene and pinene degradation, and carotenoid biosynthesis. The activation or up-regulation of various genes and pathways associated with stress resistance demonstrated that the resistant cultivar GC-4 displayed enhanced defense mechanisms against Foc. These findings provide valuable insights into the defense responses of cumin that could contribute to the development of cumin cultivars with improved resistance against Foc.

Integrative miRNA-mRNA network analysis to identify crucial pathways of salinity adaptation in brain transcriptome of Labeo rohita.

Introduction: Brain being the master regulator of the physiology of animal, the current study focuses on the gene expression pattern of the brain tissue with special emphasis on regulation of growth, developmental process of an organism and cellular adaptation of Labeo rohita against unfavourable environmental conditions. Methods: RNA-seq study was performed on collected brain samples at 8ppt salt concentration and analyzed for differential gene expression, functional annotation and miRNA-mRNA regulatory network. Results: We found that 2450 genes were having significant differential up and down regulation. The study identified 20 hub genes based on maximal clique centrality algorithm. These hub genes were mainly involved in various signaling pathways, energy metabolism and ion transportation. Further, 326 up and 1214 down regulated genes were found to be targeted by 7 differentially expressed miRNAs i.e., oni-miR-10712, oni-miR-10736, ssa-miR-221-3p, ssa-miR-130d-1-5p, ssa-miR-144-5p and oni-miR-10628. Gene ontology analysis of these differentially expressed genes led to the finding that these genes were involved in signal transduction i.e., calcium, FOXO, PI3K-AKT, TGF-ß, Wnt and p53 signalling pathways. Differentially expressed genes were also involved in regulation of immune response, environmental adaptation i.e., neuroactive ligand-receptor interaction, ECM-receptor interaction, cell adhesion molecules and circadian entrainment, osmoregulation and energy metabolism, which are critical for salinity adaptation. Discussion: The findings of whole transcriptomic study on brain deciphered the miRNA-mRNA interaction patterns and pathways associated with salinity adaptation of L. rohita.

Kidney transcriptome response to salinity adaptation in Labeo rohita.

The increasing salinization of freshwater resources, owing to global warming, has caused concern to freshwater aquaculturists. In this regard, the present study is aimed at economically important freshwater fish, L. rohita (rohu) adapting to varying degrees of salinity concentrations. The RNA-seq analysis of kidney tissue samples of L. rohita maintained at 2, 4, 6, and 8 ppt salinity was performed, and differentially expressed genes involved in various pathways were studied. A total of 755, 834, 738, and 716 transcripts were downregulated and 660, 926, 576, and 908 transcripts were up-regulated in 2, 4, 6, and 8 ppt salinity treatment groups, respectively, with reference to the control. Gene ontology enrichment analysis categorized the differentially expressed genes into 69, 154, 92, and 157 numbers of biological processes with the p value < 0.05 for 2, 4, 6, and 8 ppt salinity groups, respectively, based on gene functions. The present study found 26 differentially expressed solute carrier family genes involved in ion transportation and glucose transportation which play a significant role in osmoregulation. In addition, the upregulation of inositol-3-phosphate synthase 1A (INO1) enzyme indicated the role of osmolytes in salinity acclimatization of L. rohita. Apart from this, the study has also found a significant number of genes involved in the pathways related to salinity adaptation including energy metabolism, calcium ion regulation, immune response, structural reorganization, and apoptosis. The kidney transcriptome analysis elucidates a step forward in understanding the osmoregulatory process in L. rohita and their adaptation to salinity changes.

Magnetization of a warm plasma by the nonstationary ponderomotive force of an electromagnetic wave.

It is shown that magnetic fields can be generated in a warm plasma by the nonstationary ponderomotive force of a large-amplitude electromagnetic wave. In the present Brief Report, we derive simple and explicit results that can be useful for understanding the origin of the magnetic fields that are produced in intense laser-plasma interaction experiments.

Utility of cysticercus fasciolaris antigen in Dot ELISA for the diagnosis of neurocysticercosis.

BACKGROUND: Clinical diagnosis of neurocysticercosis (NC) is established by CT scan and MRI. However, absolute diagnosis is not possible in a fair number of cases, and serological assays are used as adjunct. Besides, CT scan and MR imaging are resource-intensive tests and not practical for screening in endemic areas. AIM: To provide a low-cost, efficient, and reproducible assay for the detection of antibodies against cysticerci. Hence we have attempted to standardize and evaluate the diagnostic utility of the cysticercus fasciolaris antigen in a Dot ELISA assay for diagnosis of NC. SETTING AND DESIGN: Tertiary hospital-based, case-control series. MATERIALS AND METHODS: Confirmed cases of NC diagnosed by presence of ring lesions in CT scan or MR imaging with presence of scolex were taken as positive controls (n = 50). Negative controls (n = 50) included subjects with normal CT scan studies (n = 30) and diseased controls with ring lesions in CT scan confirmed to be neurotuberculosis (n = 20). Dot ELISA was standardized and validated with commercially available ELISA (UBI, USA) using sera from the study groups. STATISTICAL ANALYSIS: Chi-square test was used to compare the immunodiagnostic performance of the two tests. P value less than .05 (P < 0.05) was considered significant. RESULTS: The Dot ELISA had a sensitivity of 88% and specificity of 74% with a positive predictive value of 77.19% and negative predictive value of 81.06%. Likelihood ratios for a positive and a negative test were 3.4 and 0.2. The sensitivity and specificity of commercial ELISA were 92% and 84% respectively. Difference between the performances of the two tests was not significant statistically. CONCLUSIONS: Dot ELISA has sensitivity and specificity comparable to ELISA for the diagnosis of NC. The test is simpler, not requiring expertise and instrumentation. Further validation of the test as a screening tool is required.

Comparisons between scolex and membrane antigens of Cysticercus fasciolaris and Cysticercus cellulosae larvae for immunodiagnosis of neurocysticercosis.

The antigen source for enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunotransfer blot for neurocysticercosis is generally Taenia solium. A comparison of the membrane and scolex extracts of Cysticercus cellulosae and Cysticercus fasciolaris (larval stage of Taenia taeniaeformis) for the immunodiagnosis of neurocysticercosis has been performed. C. fasciolaris cysts were produced experimentally in rat liver. C. cellulosae was obtained from muscle of infected pigs. The antigen extracts of membrane and scolex were compared using ELISA in 50 patients and 50 control participants to detect immunoglobulin (Ig) G or IgM antibodies. Proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were immunoprobed using pooled and individual sera. The gold standard for diagnosis was visualization of scolex in ring lesions by magnetic resonance imaging or computed tomography scans. ELISA for IgG antibodies using C. fasciolaris membrane had the highest sensitivity of 94%. Specificity ranged from 78% to 90%. Immunoreactive bands common to all 4 antigens were seen between 60 and 70 kDa and 40 and 45 kDa. The presence of comparative antigenic bands between human and rat pathogens provides convincing evidence for use of C. fasciolaris antigens for immunodiagnostic procedures. The antigen can be produced in small animals in standardized laboratory conditions within 60 days.

ELISA in the evaluation of therapeutic response to albendazole in neurocysticercosis.

BACKGROUND: Immunological tests are frequently used in the diagnosis of neurocysticercosis (NC), but scant literature is available on the efficacy of these tests in the assessment of therapeutic response. An ELISA using the Cysticercus fasciolaris larval stage of T. taeniaeformis has been evaluated in the post-treatment follow-up of NC in a cohort of 165 cases. METHODS: Cases (n=165) with at least one active cyst documented by computed tomography and a positive pre-treatment serum ELISA for IgG and/or IgM antibodies were treated with albendazole. CT scan and ELISA tests were repeated at 6 months in 148 cases who returned for follow-up. RESULTS: A radiological response was observed in 132 of 148 cases at follow-up. Sixteen cases were non-responders. Amongst the responders, 111/128 (IgG) and 93/117 (IgM) respectively had converted to negative antibody titers at 6 months. Thirteen of 16 and 12 of 15 non-responders continued to show high anti-Cysticercus IgG and IgM titers. IgG ELISA, IgM ELISA and combined IgG and IgM results exhibited a sensitivity (%) of 81.3, 80.0 and 100, a specificity (%) of 86.7, 79.5 and 72.0, a positive predictive value (PV%) of 97.4, 96.9 and 30.2, and a negative PV(%) of 97.4, 96.9, 100 respectively. CONCLUSION: IgG ELISA is a sensitive and specific tool to assess treatment response. A negative ELISA result for both IgG and IgM antibodies denotes a cure. While ELISA cannot replace the visual confirmation provided by radiological imaging in follow-up, the addition of an ELISA test may help overcome the limitations in interpretation of CT scans.