JNJ-26366821, a novel thrombopoietin mimetic peptide (TPOm), is shown to increase platelets (PLTs) transiently in peripheral blood. We hypothesized that increases in PLT counts may involve stimulation of hematopoiesis via induction of cytokines, growth factors, and microRNAs. Hence, we measured various cytokines, chemokines, and growth factors in serum. Time-course analysis of G-CSF, IL-5, IL-6, IL-9, IL-10, TNFα, IL-1α, and IL-1β expression was significantly altered in the control group at 9.5 Gy compared to a lower non-lethal dose of 7 Gy on days 7 to 15 post-exposure. TPOm pre-treatment significantly ameliorated the changes in expression of these pro-inflammatory cytokines and growth factors. Additionally, we show that TPOm differentially modulates the miRNA expression profiles in the spleen of irradiated mice compared to controls at both early times as well as later times after irradiation. These results suggest a possible role of TPOm in protecting animals from radiation-induced thrombocytopenia and lethality by attenuating radiation-induced inflammatory cytokines and miRNAs.

ROCK1 plays an important role in phagocytosis by inducing cytoskeletal rearrangement. Although the transcriptional regulation of ROCK1 is known but its post-transcriptional regulation is underexplored. We intended to find a mechanism of microglial phagocytosis through possible post-transcriptional regulation of ROCK1. The study identified miR-129-5p as a regulator of microglial phagocytosis following exposure to an environmental stressor, arsenic, combining in silico analysis, mutational analysis, in vitro experiments, and validation in BALB/c mouse. The in silico analysis and in vitro studies with mouse primary neonatal microglia, BV2 microglia, ex vivo microglia, and human microglial cell line CHME3 revealed that arsenic exposure increases microglial phagocytosis. Arsenic exposure was also observed to increase the level of miR-129-5p and consequently decrease the level of ROCK1 protein. In vitro experiments and mutational analysis confirmed the in silico predicted binding site of miR-129-5p on the 3'UTR of ROCK1 and also confirmed the shuttling of ROCK1 mRNA into the cytoplasmic-processing body (p-body) in mouse microglia. Downstream to ROCK1, Rac1 has also been studied to pinpoint the partners in the signaling axis. The role of miR-129-5p in microglial phagocytosis was studied in vitro and validated in vivo in BALB/c mouse by stereotactically injecting anti-miR-129-5p and assessing the phagocytosis in ex vivo microglia and colocalization of Iba1 and PSD95 in brain cryosection. Finally, experiments with arsenic, anti-miR-129-5p, ROCK1 & Rac1 siRNA in various combinations confirmed the miR-129-5p→ROCK1→Rac1→Phagocytosis signaling axis. Overall, the study revealed miR-129-5p as an important regulator of microglial phagocytosis with potential implication in synaptic plasticity and neurodegenerative complications.

Coronavirus disease 2019 (COVID-19) is a deadly human viral disease with a high rate of infection, morbidity, and mortality. Although vaccines and antiviral treatments are available, hospitalizations remain steady, and concerns about long-term consequences persist. Therefore, there is a great urgency to develop novel therapies. Here, we analyzed the role of miR-155, one of the most powerful drivers of host antiviral responses including immune and inflammatory responses, in the pathogenicity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Endogenous microRNAs (miRNAs, miRs) are key molecules in preventing viral entry and replication while building an antiviral cellular defense. Our study reveals that miR-155 expression is elevated in patients with COVID-19. Using a mouse model transgenic for human angiotensin-converting enzyme receptor 2, we evaluated the potential of anti-miR-155 therapy. Treating SARS-CoV-2-infected mice with anti-miR-155 significantly reduced miR-155 expression, improved survival, and slightly increased body weight. Notably, these mice showed altered expression of cytokines in the lungs. These findings suggest anti-miR-155 could be a promising therapy to mitigate the cytokine storm and long-lasting symptoms induced by SARS-CoV-2 infection, improving public health outcomes and enhancing global pandemic preparedness.

The detrimental effects of high -dose ionizing radiation on human health are well-known, but the influence of sex differences on the delayed effects of acute radiation exposure (DEARE) remains unclear. Here, we conducted six-month animal experiments using escalating radiation doses (7-9 Gy) on male and female C57BL/6 mice. The results show that female mice exhibited greater resistance to radiation, showing increased survival at six months post -total body irradiation. LD50/30 (lethal dose expected to cause 50% lethality in 30 days) for female mice is 8.08 Gy, while for male mice it is 7.76 Gy. DEARE causes time- and sex -dependent dysregulation of microRNA expression, processing enzymes, and the HOTAIR regulatory pathway. Differential regulation of molecular patterns associated with growth, development, apoptosis, and cancer is also observed in male and female mice. These findings shed light on the molecular basis of age and sex differences in DEARE response and emphasize the importance of personalized medicine for mitigating radiation -induced injuries and diseases.

The coordination of cellular biological processes is regulated in part via metabolic enzymes acting to match cellular metabolism to current conditions. The acetate activating enzyme, acyl-coenzyme A synthetase short-chain family member 2 (Acss2), has long been considered to have a predominantly lipogenic function. More recent evidence suggests that this enzyme has regulatory functions in addition to its role in providing acetyl-CoA for lipid synthesis. We used Acss2 knockout mice (Acss2−/−) to further investigate the roles this enzyme plays in three physiologically distinct organ systems that make extensive use of lipid synthesis and storage, including the liver, brain, and adipose tissue. We examined the resulting transcriptomic changes resulting from Acss2 deletion and assessed these changes in relation to fatty acid constitution. We find that loss of Acss2 leads to dysregulation of numerous canonical signaling pathways, upstream transcriptional regulatory molecules, cellular processes, and biological functions, which were distinct in the liver, brain, and mesenteric adipose tissues. The detected organ-specific transcriptional regulatory patterns reflect the complementary functional roles of these organ systems within the context of systemic physiology. While alterations in transcriptional states were evident, the loss of Acss2 resulted in few changes in fatty acid constitution in all three organ systems. Overall, we demonstrate that Acss2 loss institutes organ-specific transcriptional regulatory patterns reflecting the complementary functional roles of these organ systems. Collectively, these findings provide further confirmation that Acss2 regulates key transcription factors and pathways under well-fed, non-stressed conditions and acts as a transcriptional regulatory enzyme.

Risks of radiation exposure necessitate the development of radioprophylactic drugs. We have reported the efficacy of CDX-301, a recombinantly developed human protein form of Fms-related tyrosine kinase 3 ligand (Flt3L), as a radioprophylactic and radiomitigatory agent. Here, we performed global microRNA profiling to further understand the mechanism of action of CDX-301. We find that CDX-301 administration 24 h prior to total body irradiation prevents radiation-induced dysregulation of microRNA biogenesis and expression in murine serum and spleen samples in a time- and tissue-dependent manner. Further analysis shows that activation of the HOTAIR regulatory pathway has a prominent function in radiationinduced injury responses, which is inhibited by pre-treatment with CDX-301. Moreover, CDX-301 attenuates radiationinduced dysregulation of several cellular functions such as inflammatory and immune responses. In corroboration, we also find that pre-treatment with CDX-301 restores the expression of bone marrow aplasia markers and inflammatory cytokines and growth factors, as well as the expression of genes associated with MAP kinase and TGF-bpathways that are altered by radiation. Our findings provide new insights into point to a possible novel radioprotective drug for the prevention of irradiation-induced injury and hematopoietic acute radiation syndrome.

Inflammation has recently achieved remarkable consideration due to the high severity, frequency, and economic burden of inflammatory diseases. Although acute inflammation is an essential host defense response to counteract diverse harmful stimulus, to speed up recovery of damaged cells/tissues, and maintain homeostasis, chronic inflammation may lead to the development and progression of various diseases, including allergic, respiratory, cardiovascular, autoimmune, cancer, and other diseases. It is essential to control dysregulated inflammatory responses and prevent cell/tissue damage. Therefore, it is of utmost importance to uncover the possible regulators of inflammatory mechanisms. In this context, microRNAs (miRNAs) that play critical regulatory roles in almost every cellular and developmental process are emerging as fine-tuned signaling regulators to modulate inflammatory responses and prevent or treat inflammatory diseases. Here, we provide a broad overview of miRNAs and their regulatory mechanisms in inflammatory diseases. Furthermore, the role of miRNAs as biomarkers and antiinflammatory agents leads to new avenues in the diagnosis and prevention or treatment of inflammatory diseases.

ABSTRACT Coronavirus disease 2019 (COVID-19) is a viral illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and is a worsening global pandemic. COVID-19 has caused at least 1.7 million deaths worldwide and over 300,000 in the United States. Recently, two promising vaccines are being administered in several countries. However, there remains an urgent need for a therapeutic treatment for COVID-19 patients with severe respiratory damage that can lead to intensive care, prolonged hospitalization, or mortality. Moreover, an increasing population of patients manifest lingering disabling symptoms (called Long Haulers). Here, we tested the efficacy of a recombinant neural epidermal growth factor like 1 protein variant (NELL1-NV1) in a COVID-19 mouse model, transgenic mice expressing the human angiotensin I-converting enzyme 2 (ACE2) receptor (tg-mice hACE2) infected with SARS-CoV-2. The administration of NELL1-NV1 to SARS-CoV-2-infected tg-mice hACE2 significantly improved clinical health score and increased survival. Analyses of bronchoalveolar (BAL) fluid demonstrated decreased levels of several cytokines and chemokines (IFN-γ, IL-10, IL-12 p70, CXCL-10/IP-10, MIG and Rantes), in NV1-treated treated mice compared to controls. Cytokines including IL-1α, IL-9, IL-6, LIX/CXCL5, KC/CXCL1, MIP-2/CXCL2, MIP-1α/CCL3, and G-CSF, critical to immune responses such as neutrophil recruitment, viral clearance and vascularization, were increased compared to controls. Our data suggest the potential of NELL1-NV1-based therapy to mitigate the cytokine storm, modulate the abnormal immune response and repair respiratory tissue damage in COVID-19 patients.

With the advancement in transcriptomic technology, it is now possible to get insights into undescribed phenomena of the continually evolving arms race of plant–pathogen interactions. The study of this interaction describes various mechanisms such as the plant defense pathway, regulatory gene annotation, gene network, and biochemical metabolism of plants and pathogens, which still remain a major challenge in agriculture. Researchers can understand the mechanisms of altering gene expression during compatible and incompatible interaction of pathogenic organisms at the molecular level. Transcriptome profiling could efficiently identify the differential changes in gene expression in response to infection, and thereby provide information regarding the basal defense system, resistance, pathogenicity of plant pathogens, and the difference between host and nonhost defense mechanisms of plants. Based on this information, several broad spectrum disease-resistant crop plants have been generated.

Non-coding RNAs (ncRNAs), notably microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), have recently gained increasing consideration because of their versatile role as key regulators of gene expression. They adopt diverse mechanisms to regulate transcription and translation, and thereby, the function of the protein, which is associated with several major biological processes. For example, proliferation, differentiation, apoptosis, and metabolic pathways demand fine-tuning for the precise development of a specific tissue or organ. The deregulation of ncRNA expression is concomitant with multiple diseases, including lung diseases. This review highlights recent advances in the post-transcriptional regulation of miRNAs and lncRNAs in lung diseases such as asthma, chronic obstructive pulmonary disease, cystic fibrosis, and idiopathic pulmonary fibrosis. Further, we also discuss the emerging role of ncRNAs as biomarkers as well as therapeutic targets for lung diseases. However, more investigations are required to explore miRNAs and lncRNAs interaction, and their function in the regulation of mRNA expression. Understanding these mechanisms might lead to early diagnosis and the development of novel therapeutics for lung diseases.

Cystic fibrosis (CF) occurs as a result of mutations in the cystic fibrosis transmembrane conductance regulator ( CFTR ) gene, which lead to misfolding, trafficking defects, and impaired function of the CFTR protein. Splicing factor proline/glutamine-rich (SFPQ) is a multifunctional nuclear RNA-binding protein (RBP) implicated in the regulation of gene expression pathways and intracellular trafficking. Here, we investigated the role of SFPQ in the regulation of the expression and function of F508del-CFTR in CF lung epithelial cells. We find that the expression of SFPQ is reduced in F508del-CFTR CF epithelial cells compared to WT-CFTR control cells. Interestingly, the overexpression of SFPQ in CF cells increases the expression as well as rescues the function of F508del-CFTR. Further, comprehensive transcriptome analyses indicate that SFPQ plays a key role in activating the mutant F508del-CFTR by modulating several cellular signaling pathways. This is the first report on the role of SFPQ in the regulation of expression and function of F508del-CFTR in CF lung disease. Our findings provide new insights into SFPQ-mediated molecular mechanisms and point to possible novel epigenetic therapeutic targets for CF and related pulmonary diseases.

Endophytes are microorganisms residing inside the plants with an escalating interest for the scientific community. The plethora of endophyte-derived medicinal molecules and the application potentials in agriculture and bioremediation are some of the reasons why endophytes are extensively studied. Endophytes which can be either endophytic fungi or bacteria have been isolated from a wide variety of plants. The huge diversity among the endophytic microbes and the host plants hinders the establishment of a uniform protocol for isolation of endophytes. However, the most common technique that is currently used involves a thorough surface sterilization followed by fragmentation of the plant tissue and culture of the fragments onto agar plates. Nevertheless, non-culturable microorganisms are usually omitted from the aforementioned procedure and molecular approaches have to be applied for a more inclusive isolation. Sequencing of 16S rRNA for bacteria and ITS or 18S rRNA for fungi can be applied to both culturable and unculturable endophytes. Sometimes, morphological examination can be a further validation for the identification of the isolated strains. Subsequently, a variety of biochemical and molecular approaches including colorimetry, spot assays, chromatographies, and gene-targeted PCR (polymerase chain reaction) amplification are performed in order to further characterize the endophytic strains. Scrutinous functional analysis of endophytes may provide valuable insights into the advancement of agriculture, medicine, and industry. In this chapter, methods for the isolation of endophytic bacteria/fungi, identification, and their characterization depending on their functional role are discussed and provided as a helpful toolbox for a new researcher in the endophyte field.

Nutrient procurement specifically from nutrient-limiting environment is essential for pathogenic bacteria to survive and/or persist within the host. Long-term survival or persistent infection is one of the main reasons for the overuse of antibiotics, and contributes to the development and spread of antibiotic resistance. Mycobacterium tuberculosis is known for long-term survival within the host, and develops multidrug resistance. Before and during infection, the pathogen encounters various harsh environmental conditions. To cope up with such nutrient-limiting conditions, it is crucial to uptake essential nutrients such as ions, sugars, amino acids, peptides, and metals, necessary for numerous vital biological activities. Among the various types of transporters, ATP-binding cassette (ABC) importers are essentially unique to bacteria, accessible as drug targets without penetrating the cytoplasmic membrane, and offer an ATP-dependent gateway into the cell by mimicking substrates of the importer and designing inhibitors against substrate-binding proteins, ABC importers endeavour for the development of successful drug candidates and antibiotics. Alternatively, the production of antibodies against substrate-binding proteins could lead to vaccine development. In this review, we will emphasize the role of M. tuberculosis ABC importers for survival and virulence within the host. Furthermore, we will elucidate their unique characteristics to discover emerging therapies to combat tuberculosis.

Coronavirus disease 2019 (COVID-19) is a recent global pandemic. It is a deadly human viral disease, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with a high rate of infection, morbidity and mortality. Therefore, there is a great urgency to develop new therapies to control, treat and prevent this disease. Endogenous microRNAs (miRNAs, miRs) of the viral host are key molecules in preventing viral entry and replication, and building an antiviral cellular defense. Here, we have analyzed the role of miR-155, one of the most powerful drivers of host antiviral responses including immune and inflammatory responses, in the pathogenicity of SARS-CoV-2 infection. Subsequently, we have analyzed the potency of anti-miR-155 therapy in a COVID-19 mouse model (mice transgenic for human angiotensin I-converting enzyme 2 receptor (tg-mice hACE2)). We report for the first time that miR-155 expression is elevated in COVID-19 patients. Further, our data indicate that the viral load as well as miR-155 levels are higher in male relative to female patients. Moreover, we find that the delivery of anti-miR-155 to SARS-CoV-2-infected tg-mice hACE2 effectively suppresses miR-155 expression, and leads to improved survival and clinical scores. Importantly, anti-miR-155-treated tg-mice hACE2 infected with SARS-CoV-2 not only exhibit reduced levels of pro-inflammatory cytokines, but also have increased anti-viral and anti-inflammatory cytokine responses in the lungs. Thus, our study suggests anti-miR-155 as a novel therapy for mitigating the lung cytokine storm induced by SARS-CoV-2 infection.

Human listeriosis caused by Listeria monocytogenes (LM) is a serious public health concern. The disease leads to several health issues like abortion, stillbirth, septicaemia, meningitis and meningoencephalitis. The main targets are pregnant women, neonates, elderly or immunocompromised people 1 .

The early detection of Listeria monocytogenes (L. monocytogenes) and understanding the disease burden is of paramount interest. The failure to detect pathogenic bacteria in the food industry may have terrible consequences, and poses deleterious effects on human health. Therefore, integration of methods to detect and trace the route of pathogens along the entire food supply network might facilitate elucidation of the main contamination sources. Recent research interest has been oriented towards the development of rapid and affordable pathogen detection tools/techniques. An innovative and new approach like biosensors has been quite promising in revealing the foodborne pathogens. In spite of the existing knowledge, advanced research is still needed to substantiate the expeditious nature and sensitivity of biosensors for rapid and in situ analysis of foodborne pathogens. This review summarizes recent developments in optical, piezoelectric, cell-based, and electrochemical biosensors for Listeria sp. detection in clinical diagnostics, food analysis, and environmental monitoring, and also lists their drawbacks and advantages.

Listeria monocytogenes has emerged as the deadly pathogen inflicting high mortality in humans and animals. To investigate the strain-specific characteristics of L. monocytogenes, and their role(s) in virulence and ecological sustenance, we sequenced genome of three L. monocytogenes strains BHU1, 2 and 3 isolated from the Ganges river, agricultural soil and human placenta bit, respectively, and compared it with L. monocytogenes EGD-e serovar 1/2a and L. monocytogenes F2365 serovar 4b strain. The contigs of all the three strains had a similarity (>90%) in regions that aligned with EGD-e and F2365. A total of 2872 core genes on the set of strains were identified in BHU1, 2 and 3 strains. In the mice virulence assay, BHU2 and 3 strains showed pathogenicity while BHU1 was non-pathogenic. These strains were also characterized with unique genes (8 genes in BHU1, 12 in BHU 2, and 17 in BHU 3 strain). Phylogenetic analysis based on multilocus sequence typing revealed BHU1, 2 and 3 strains to be more closely related to lineage I, serotype 4b, clonal complex 1, and sequence type 328. BHU1 strain seemed to harbor nucleotide mutation from A to G in the major virulence genes i.e. hemolysin D and listeriolysin O. Strain-specific mutations 8, 5 and 2 were identified in BHU1, 2 and 3, respectively, compared to F2365. Though all the three strains were genetically very close, the observed differences may play the crucial role(s) in their virulence attributes, and also, in the prevalence of L. monocytogenes.

Listeria infection is major health problem causing listeriosis that manifests as abortion, stillbirth, septicemia, meningitis and meningoencephalitis. Listeriolysin 0 is the cholesterol-dependent cytolysin toxin involved in the escape of L. monocytogenes from primary and secondary intracellular vacuoles and, therefore, can serve as the vital target for vaccine development. Consequently, the present study was aimed to design epitope-based vaccine against Listeria. LLO, ILO, and SLO proteins from L. monocytogenes, L. ivanovii and L. seeligeri, respectively were analyzed using various bioinformatics and immuonoinformatics tools, including sequence and structure-based ones. A total of 11 antigenic B-cell epitopes, and 4 and 3 allelic classes for MHC class I and MHC class II binding peptides, respectively were predicted for LLO protein. The unique peptide (363)LGDLRD(368) was identified in the LLO protein. Further, we also observed that IgG class of B-cells were predominant in these proteins. The study revealed potential B-cell and T-cell epitope that can raise the desired immune response against these proteins. The present study would, therefore, be helpful in designing and predicting novel vaccine candidates, which in near future might offer the source for eradicating listeriosis.

ABSTRACT We present here the whole-genome sequences of Listeria monocytogenes from Ganges River water, agricultural soil, and human clinical samples from Varanasi, India, which will be used for a comparative analysis.