Correlation Between Estrogen Receptor α Gene Polymorphism (c454-397T>C) with Serum Estradiol Levels and Known Risk Factors in Patients with Myocardial Infarction.

Myocardial infarction (MI) remains the most common cause of cardiac failure and continuous increasing rate of morbidity and mortality. We aimed to investigate the association of estrogen receptor-α (ESR1) gene polymorphism c454-397T>C with serum estradiol levels and dyslipidemia in 220 patients with MI in the age range of 35-70 years of both the genders. Genotyping study was performed through PCR-RFLP method using PvuII restriction enzyme. Serum estradiol level was estimated using the Access Sensitive Estradiol assay kit. Men patients had 43.2% increased risk for TC heterozygote in co-dominant (OR 10.66) and over-dominant models (OR 8.30), while women patients had 50% increased risk in co-dominant (OR 16.57) and over-dominant (OR 14.04) models. Variant C allele showed 25% increased risk of MI for in men (OR 2.24; CI 1.49-3.36; p = 0.0001), and 24% increased risk in women (OR 3.35; CI 1.95-5.76; p = 0.0001). Men patients had significantly increased serum estradiol levels compared to controls (25.28 ± 5.80 vs 17.04 ± 2.01; p < 0.0001). Significant difference was observed in estradiol levels between men and women patients (25.28 ± 5.80 vs 17.56 ± 3.32; p < 0.0001). Furthermore, significantly increased estradiol level was found in men patients compared to women for TT (25.46 ± 5.91 vs 16.71 ± 4.46; p < 0.0001), and TC genotypes (25.47 ± 5.91 vs 17.70 ± 2.86; p < 0.0001). Significantly increased HDL levels were observed in men patients with TC (43.10 ± 8.18 vs 38.91 ± 7.84; p < 0.01) and CC (47.16 ± 8.09 vs 38.91 ± 7.84; p < 0.001) genotypes compared to TT genotype. These findings suggest that TC heterozygote plays an important role as a genetic risk factor during MI pathogenesis in the South Indian population. Supplementary Information: The online version contains supplementary material available at 10.1007/s12291-022-01104-1.

Association of Angiotensin II Type 1 Receptor (AT1R) Gene Polymorphism with Angiotensin II Serum Levels in Patients with Essential Hypertension.

Essential hypertension (EH) is a multifactorial, polygenic condition, and is one of the most important comorbidities that contributes to stroke, myocardial infarction, cardiac failure, and renal failure. The continuous increasing rate of morbidity and mortality associated with EH presents an unmet need of population-based studies to explore pathophysiology as well as newer strategies for better diagnosis, prognosis and treatment. This study aimed to determine genotype and allele frequencies of A1166C polymorphism of AT1R gene in Indian patients with EH and correlated with serum levels of Angiotensin II. A total of 200 patients with EH and 200 age- and gender-matched control individuals were included in this study from the General Medicine Department Outpatient at Narayana Medical College and Hospital, Nellore, Andhra Pradesh, India. Patients with systolic blood pressure (SBP) ≥ 140 mmHg and/or diastolic blood pressure (DBP) ≥ 90 mmHg were considered as hypertensive. The findings of this study revealed significantly increased risk of C/A heterozygote and allele C in both men and women. Moreover, both men and women patients with EH showed higher serum levels of Angiotensin II with C/A as well as AA genotypes. These findings indicate a significant association of 1166 C/A polymorphism of the AT1R gene with increased risk of hypertension in Indian population.

Antimicrobial, antioxidant, antiviral activity, and gas chromatographic analysis of Varanus griseus oil extracts.

There is an urgent need to develop natural antimicrobials for the control of rapidly mutating drug-resistant bacteria and poultry viruses. Five extracts were prepared using diethyl ether, ethyl acetate, methanol, 1-butanol and n-hexane from abdominal fats of Varanus griseus locally known as Indian desert monitor. Antibacterial, antioxidant and antiviral activities from oil extracts were done through disc diffusion method, stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay and in ovo antiviral assay, respectively. The gas chromatography mass spectrometry (GC-MS) analyses were used to determine principal active compounds and chemical profile of each oil extract. n-Hexane extract showed clear zones of inhibition (ZOI) against Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae (12 ± 0.5 mm, 9 ± 0.5 mm, and 9 ± 0.5 mm) while diethyl ether extract exhibited significant antibacterial activity (11 ± 0.5 mm) against Proteus vulgaris only. In case of drug-resistant strains, methanol extract was active (6 ± 0.5 mm) against Staphylococcus aureus, whereas n-hexane extract has shown ZOI 11 ± 0.5 mm against P. aeruginosa. Range of percentage scavenging activity of V. griseus oil extracts from DPPH free radical assay was 34.9-70.7%. For antiviral potential, growth of new castle disease virus (NDV) was effectively inhibited by all five extracts (HA titer = 0-4). The highest antiviral activity against avian influenza virus (H9N2) was observed from methanol, diethyl ether and 1-Butanol oil extracts with HA titers of 2, 2 and 0, respectively. Methanol, diethyl ether, 1-butanol and n-hexane oil extracts produced best hemagglutination assay (HA) titer values (0, 0, 4 and 0) against infectious bronchitis virus (IBV). Ethyl acetate and 1-Butanol extract exhibited good antiviral potential against infectious bursal disease virus (IBDV) with indirect hemagglutination assay (IHA) titers of 8 and 4, respectively. Main classes of identified compounds through gas chromatography were aldehydes, fatty acids, phenols and esters. GC-MS identified 11 bioactive compounds in V. griseus oil extracts. It is summarized that V. griseus oil has strong antioxidant activity and good antimicrobial potential because of its bioactive compounds.

Status of Vitamin D Receptor Gene Polymorphism and 25-Hydroxy Vitamin D Deficiency with Essential Hypertension.

Essential hypertension (EH) is a multifactorial and complex disease with high rate of incidence and associated co-morbidities. Previous studies do not provide unanimous results for the risk of hypertension and association with Fok I genotype frequency and serum vitamin D levels. Hence, this study was undertaken to determine the status of Fok I vitamin D receptor (VDR) gene polymorphism along with vitamin D levels and blood pressure in patients with EH. Four hundred (200 controls and 200 cases of essential hypertension) participants from general Indian population were enrolled in this study. Peripheral blood samples were collected for genotyping Fok I-VDR gene polymorphism using PCR-RFLP method whereas 25-OH vitamin D levels in serum were quantified using high performance liquid chromatography (HPLC). Significantly reduced 25-OH vitamin D levels were observed in patients with EH (24.04 ± 8.62 vs 50.46 ± 15.46) compared to control subjects (p = 0.0001). Homozygous recessive genotype 'ff' frequency was increased by 8.06 fold (CI: 3.71-17.47, p = 0.0001) in patients with EH compared to dominant 'FF' genotype frequency. In conclusion, recessive 'ff' genotype frequency correlates with reduced serum vitamin D levels and results in significantly increased systolic and diastolic blood pressures leading to predisposition of EH.

Amino acid starvation sensing dampens IL-1ß production by activating riboclustering and autophagy.

Activation of the amino acid starvation response (AAR) increases lifespan and acute stress resistance as well as regulates inflammation. However, the underlying mechanisms remain unclear. Here, we show that activation of AAR pharmacologically by Halofuginone (HF) significantly inhibits production of the proinflammatory cytokine interleukin 1ß (IL-1ß) and provides protection from intestinal inflammation in mice. HF inhibits IL-1ß through general control nonderepressible 2 kinase (GCN2)-dependent activation of the cytoprotective integrated stress response (ISR) pathway, resulting in rerouting of IL-1ß mRNA from translationally active polysomes to inactive ribocluster complexes-such as stress granules (SGs)-via recruitment of RNA-binding proteins (RBPs) T cell-restricted intracellular antigen-1(TIA-1)/TIA-1-related (TIAR), which are further cleared through induction of autophagy. GCN2 ablation resulted in reduced autophagy and SG formation, which is inversely correlated with IL-1ß production. Furthermore, HF diminishes inflammasome activation through suppression of reactive oxygen species (ROS) production. Our study unveils a novel mechanism by which IL-1ß is regulated by AAR and further suggests that administration of HF might offer an effective therapeutic intervention against inflammatory diseases.

Genotyping simplified: rationally designed antisense oligonucleotide-mediated PCR amplification-free colorimetric sensing of viral RNA in HCV genotypes 1 and 3.

Molecular diagnosis of viral genotyping devoid of polymerase chain reaction (PCR) amplification in clinical cohorts has hitherto been challenging. Here we present a simplified molecular diagnostic strategy for direct genotyping of hepatitis C virus (HCV) 1 and 3 (prevalent worldwide) using a combination of rationally designed genotype-specific antisense oligonucleotides (ASOs) and plasmonic gold nanoparticles. The ASOs specific to genotypes 1 and 3 have been designed from the nonstructural region 5A (NS5A) of the viral genome using the ClustalW multiple sequence alignment tool. A total of 79 clinical samples including 18 HCV genotype 1, 18 HCV genotype 3, one HIV positive, one HBV positive, and 41 healthy controls have been tested against both the designed ASOs. The study reveals 100% specificity and sensitivity with the employed samples and thereby opens up new avenues for PCR-free direct genotyping of other viruses as well, through the rational design of ASOs.

Activation of integrated stress response pathway regulates IL-1ß production through posttranscriptional and translational reprogramming in macrophages.

Immune cells sense and programme its cellular machinery appropriately to the environmental changes through the activation of cytoprotective adaptive pathway so-called the "integrated stress response (ISR)". However, the mechanisms implicated in ISR-induced protective responses are poorly understood. Here, we show that ISR activation by arsenite (Ar) results in suppression of IL-1ß production in macrophages and inhibition of DSS-induced colitis in a murine model through a novel posttranscriptional and translation regulatory (PTR) mechanism. Ar triggers PTR events through eIF2α-phosphorylation, which results in the attenuation of active polysome formation leading to the accumulation of translationally stalled IL-1ß mRNAs. Translationally stalled IL-1ß mRNAs recruit RNA-binding proteins (TIA-1/TIAR), resulting in the formation of RBP-RNA complexes known as stress granules (SGs). The SGs bound IL-1ß mRNAs might undergo degradation through induction of autophagy. Also, we show that Ar posttranslationally impairs processing and secretion of IL-1ß by diminishing inflammasome activation. Altogether, this study unveils a novel mechanism of IL-1ß regulation and further suggests that pharmacological activation of cytoprotective ISR pathway might provide an effective therapeutic intervention against inflammatory diseases.

Transcriptome meta-analysis identifies immune signature comprising of RNA binding proteins in ulcerative colitis patients.

Ulcerative colitis (UC) is a persistent inflammatory illness, which is clinically categorised as Inflammatory bowel disease (IBD), affecting millions of people worldwide. The precise cause behind the pathology of the disease remains unknown. However, the involvement of multiple factors including genetic predisposition, immunological deregulations, microbiota imbalance, and environmental triggers has been suggested. Amongst all these factors, the over-active immunological response reported in UC patients seems to be a promising target for therapy. Moreover, identification of gene signatures associated with disease onset and progression would help in better understanding of the molecular mechanisms involved in the disease pathogenesis. Here, we have conducted meta-analysis of gene expression profiles of UC patient microarray datasets accessible in public databases and further validated the in-silico findings in UC patients' blood samples. Our study reveals that UC pathogenesis perturbs expression of several inflammatory genes. In addition, we report a novel gene signature comprising of TIA1 (T cell restricted intracellular antigen) and TIAR (TIA1 related protein; also known as TIAL1), which were found to be significantly downregulated in UC patients. TIA1 and TIAR are RNA-binding proteins (RBPs), which function as a translational represser by binding to ARE sequences in the 3' UTR of mRNAs encoding inflammatory mediators including cytokines. Our findings demonstrate that deletion of TIAR using gene specific siRNAs in-vitro results in enhanced production of inflammatory cytokine IL-1ß. In conclusion, the findings of this study reveal that down regulation of TIA1/TIAR genes could be responsible for UC associated inflammation. This study highlights the usefulness of the meta-analysis approach in the identification of unique gene signatures that might deliver mechanistic insights into UC pathogenesis and possibly assist in discovery of prognostic markers and therapeutic interventions.

Repopulation of decellularized whole organ scaffold using stem cells: an emerging technology for the development of neo-organ.

Demand of donor organs for transplantation in treatment of organ failure is increasing. Hence there is a need to develop new strategies for the alternative sources of organ development. Attempts are being made to use xenogenic organs by genetic manipulation but the organ rejection against human always has been a major challenge for the survival of the graft. Advancement in the genetic bioengineering and combination of different allied sciences for the development of humanized organ system, the therapeutic influence of stem cell fraction on the reconstitution of organ architecture and their regenerative abilities in different tissues and organs provides a better approach to solve the problem of organ shortage. However, the available strategies for generating the organ/tissue scaffolds limit its application due to the absence of complete three-dimensional (3D) organ architecture, mechanical strength, long-term cell survival, and vascularization. Repopulation of whole decellularized organ scaffolds using stem cells has added a new dimension for creating new bioengineered organs. In recent years, several studies have demonstrated the potential application of decellularization and recellularization approach for the development of functional bio-artificial organs. With the help of established procedures for conditioning, extensive stem cells and organ engineering experiments/transplants for the development of humanized organs will allow its preclinical evaluation for organ regeneration before translation to the clinic. This review focuses on the major aspects of organ scaffold generation and repopulation of different types of whole decellularized organ scaffolds using stem cells for the functional benefit and their confines.

Molecular basis of sepsis: A New insight into the role of mitochondrial DNA as a damage-associated molecular pattern.

Sepsis remains a critical challenge in the field of medicine, claiming countless lives each year. Despite significant advances in medical science, the molecular mechanisms underlying sepsis pathogenesis remain elusive. Understanding molecular sequelae is gaining deeper insights into the roles played by various damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs) in disease pathogenesis. Among the known DAMPs, circulating cell-free mitochondrial DNA (mtDNA) garners increasing attention as a key player in the immune response during sepsis and other diseases. Mounting evidence highlights numerous connections between circulating cell-free mtDNA and inflammation, a pivotal state of sepsis, characterized by heightened inflammatory activity. In this review, we aim to provide an overview of the molecular basis of sepsis, particularly emphasizing the role of circulating cell-free mtDNA as a DAMP. We discuss the mechanisms of mtDNA release, its interaction with pattern recognition receptors (PRRs), and the subsequent immunological responses that contribute to sepsis progression. Furthermore, we discuss the forms of cell-free mtDNA; detection techniques of circulating cell-free mtDNA in various biological fluids; and the diagnostic, prognostic, and therapeutic implications offering insights into the potential for innovative interventions in sepsis management.

Chemical composition and antimicrobial study of Crossobamon orientalis body oil.

Geckos and their products have been used in Asian traditional medicine. Medicinal properties of desert-dwelling Gecko species, Crossobamon orientalis remain unexplored. In this study, natural bioactive macromolecules present in oil extracted from C. orientalis (COO) and their biological activities were evaluated. Chemical constitution of COO was explored by using gas chromatography mass spectrometry. Antioxidant, antiviral, and antibacterial activities of COO extracts were assessed using various assays, including DPPH free-radical-protocol, HET-CAM method, in ovo-antiviral technique, and disc-diffusion method. GC-MS study reported 40 different compounds in COO. n-hexane and methanol extracts of COO demonstrated highest DPPH radical inhibition, with values of 70 and 63.3%, respectively. Extracts of COO in solvents, namely 1-butanol, methanol, diethyl ether, and n-hexane significantly inhibited the proliferation of four pathogenic viruses. Maximum zone of inhibition was observed for Escherichia coli (13.65 ± 0.57 mm). These findings suggest that COO possesses potent antioxidant and antimicrobial properties against viral and bacterial strains, thanks to its biologically active components having no side effects. Further studies are essential to isolate and identify individual bioactive compounds present in COO and to investigate their potential as therapeutic agents.

New insights into the intricate taxonomy and phylogeny of the Sylvia curruca complex.

We use the mitochondrial cytochrome b from 213 individuals and the three nuclear introns BRM 15, myoglobin 2 and ODC 6-7 from a smaller subsample to evaluate the taxonomy of the Lesser Whitethroat Sylvia curruca (Aves, Passeriformes, Sylviidae) complex, which has long been controversial. We sequenced type material of the taxa althaea, blythi, margelanica and minula, and used topotypical material of caucasica, chuancheica, curruca and telengitica. The nuclear introns fail to resolve the complex, but cytochrome b recovers six major clades, revealing genetically identifiable populations corresponding to previously named taxa, and we propose that the names althaea, blythi, curruca, halimodendri, margelanica and minula, respectively, should be used for these. The margelanica clade is suggested to have a more extensive distribution than previously known, including both the taxon telengitica and a population in eastern Mongolia. The taxon minula is found to have a more restricted range than generally believed, only breeding in China. According to the mitochondrial gene tree, there is a basal dichotomy, with the taxa althaea, blythi, halimodendri and margelanica being part of one clade, well separated from a clade containing curruca and minula. Dating analysis suggests that a basal divergence separating curruca and minula from the other four taxa occurred between 4.2 and 7.2 mya; these two then diverged between 2.3 and 4.4 mya. The splits between the althaea, blythi, halimodendri and margelanica lineages is inferred to have occurred later, approximately between 1.0 and 2.5 mya (all 95% HPD). The nucleotide data suggest significant departure from demographic equilibrium in blythi (clade 1a), halimodendri (clade 2a) and minula, whereas tendencies are weaker for other clades. We propose that the names althaea, blythi, curruca, halimodendri, margelanica and minula should be used for the major clades. However, whether these are treated as subspecies or species is largely a matter of species definition and is not resolved by our data.

Multilocus phylogeny of the avian family Alaudidae (larks) reveals complex morphological evolution, non-monophyletic genera and hidden species diversity.

The Alaudidae (larks) is a large family of songbirds in the superfamily Sylvioidea. Larks are cosmopolitan, although species-level diversity is by far largest in Africa, followed by Eurasia, whereas Australasia and the New World have only one species each. The present study is the first comprehensive phylogeny of the Alaudidae. It includes 83.5% of all species and representatives from all recognised genera, and was based on two mitochondrial and three nuclear loci (in total 6.4 kbp, although not all loci were available for all species). In addition, a larger sample, comprising several subspecies of some polytypic species was analysed for one of the mitochondrial loci. There was generally good agreement in trees inferred from different loci, although some strongly supported incongruences were noted. The tree based on the concatenated multilocus data was overall well resolved and well supported by the data. We stress the importance of performing single gene as well as combined data analyses, as the latter may obscure significant incongruence behind strong nodal support values. The multilocus tree revealed many unpredicted relationships, including some non-monophyletic genera (Calandrella, Mirafra, Melanocorypha, Spizocorys). The tree based on the extended mitochondrial data set revealed several unexpected deep divergences between taxa presently treated as conspecific (e.g. within Ammomanes cinctura, Ammomanes deserti, Calandrella brachydactyla, Eremophila alpestris), as well as some shallow splits between currently recognised species (e.g. Certhilauda brevirostris-C. semitorquata-C. curvirostris; Calendulauda barlowi-C. erythrochlamys; Mirafra cantillans-M. javanica). Based on our results, we propose a revised generic classification, and comment on some species limits. We also comment on the extraordinary morphological adaptability in larks, which has resulted in numerous examples of parallel evolution (e.g. in Melanocorypha mongolica and Alauda leucoptera [both usually placed in Melanocorypha]; Ammomanopsis grayi and Ammomanes cinctura/deserti [former traditionally placed in Ammomanes]; Chersophilus duponti and Certhilauda spp.; Eremopterix hova [usually placed in Mirafra] and several Mirafra spp.), as well as both highly conserved plumages (e.g. within Mirafra) and strongly divergent lineages (e.g. Eremopterix hova vs. other Eremopterix spp.; Calandrella cinerea complex vs. Eremophila spp.; Eremalauda dunni vs. Chersophilus duponti; Melanocorypha mongolica and male M. yeltoniensis vs. other Melanocorypha spp. and female M. yeltoniensis). Sexual plumage dimorphism has evolved multiple times. Few groups of birds show the same level of disagreement between taxonomy based on morphology and phylogenetic relationships as inferred from DNA sequences.

Simultaneous extraction and quantification of circulating mitochondrial and nuclear DNA using a single plasma sample to predict specific molecular diagnostic implications.

The magnitude of variations in the level of circulating mitochondrial (cir-mtDNA) and nuclear DNA (cir-ncDNA) in different diseases has indicated the need for investigating a discriminative approach for evaluating their diagnostic significance. This study reports a typical in-house process for extracting both types of cir-DNAs from a single plasma sample and assessed their usefulness in discriminating type 2 diabetes mellitus patients from healthy individuals to eliminate the prevailing dispute about their discriminative role and improve their diagnostic value. This approach offers a more precise and valuable tool for distinguishing the impact of cir-mtDNA from cir-ncDNA in diagnostic implications.

Assessment of soluble thrombomodulin and soluble endoglin as endothelial dysfunction biomarkers in seriously ill surgical septic patients: correlation with organ dysfunction and disease severity.

BACKGROUND: Sepsis, a complex condition characterized by dysregulated immune response and organ dysfunction, is a leading cause of mortality in ICU patients. Current diagnostic and prognostic approaches primarily rely on non-specific biomarkers and illness severity scores, despite early endothelial activation being a key feature of sepsis. This study aimed to evaluate the levels of soluble thrombomodulin and soluble endoglin in seriously ill surgical septic patients and explore their association with organ dysfunction and disease severity. METHODOLOGY: A case control study was conducted from March 2022 to November 2022, involving seriously ill septic surgical patients. Baseline clinical and laboratory data were collected within 24 h of admission to the Surgical Intensive Care Unit. This included information such as age, sex, hemodynamic parameters, blood chemistry, SOFA score, qSOFA score, and APACHE-II score. A proforma was filled out to record these details. The outcome of each patient was noted at the time of discharge. RESULTS: The study found significantly elevated levels of soluble thrombomodulin and soluble endoglin in seriously ill surgical septic patients. The RTqPCR analysis revealed a positive correlation between soluble thrombomodulin and soluble endoglin levels with the qSOFA score, as well as, there was a positive association between RTqPCR soluble thrombomodulin and the SOFA score. These findings indicate a correlation between these biomarkers and organ dysfunction and disease severity. CONCLUSION: The study concludes that elevated levels of soluble thrombomodulin and soluble endoglin can serve as endothelial biomarkers for early diagnosis and prognostication in seriously ill surgical septic patients.

Role of circulatory miRNA-21 and associated signaling pathways in the pathogenesis of pulmonary fibrosis among individuals recovered after COVID-19 infection.

Introduction: Currently pulmonary fibrosis in post-COVID individuals represents a crucial milieu of investigation due to long-term associated complications and worse clinical outcome. Lack of studies in Indian population confers a crucial need for elucidating possible targets and mechanisms to explore better management and outcome. Hence, this study aimed to explore the role of circulating miRNA-21 in patients from South India after COVID-19 recovery, while targeting TGF-ß signaling pathway involved in the development of pulmonary fibrosis. Methods: This prospective, single centre, hospital-based study enrolled a total of 50 participants in the age group of 50 to 60 years including 25 non-infected controls and 25 patients who were recovered after 3-6 months of COVID-19 infection and presented radiological pulmonary abnormalities. Quantification of miRNA-21 and selected gene transcripts (TGF-ß, Col1A2, Col3A1, and α-SMA) was performed in plasma samples of both patients and controls. Results: Significantly increased expression levels of miRNA-21 was observed in patient samples compared to controls (4.50 ± 1.03 vs 12.60 ± 3.52, p < 0.0001) with 72.10% sensitivity and 80.10% specificity. Further, significantly increased levels of central fibrosis regulatory gene transcript TGF-ß (0.56 ± 0.27 vs 1.83 ± 0.98), two crucial collagen transcripts Col1A2 (0.62 ± 0.19 vs 1.56 ± 1.00) and Col3A1 (0.61 ± 0.27 vs 1.54 ± 0.89), and α-SMA (0.46 ± 0.17 vs 1.20 ± 0.78) was observed in patients compared to controls. Western-blot analysis also showed almost similar observations at proteins levels. Conclusion: Circulating miRNA-21 may provide crucial insights for elucidating TGF-ß mediated pulmonary remodeling involved in the fibrosis development and achieve better clinical outcome for post-COVID patients after recovery, in real-time with high diagnostic accuracy.

A Comprehensive Stemness Gene Expression Analysis Signifies the Role of ABC Transporters and Molecular Chaperons in Determining the Fate of Human Neural Precursor Cells.

Background and Aim: The major aim of this study was to identify the most common stemness genes across different stem cell types and further validate them in human fetal subventricular zone-derived primary and cultured neural precursor cells (NPCs). This study involved the use of a unique method of stemness meta-analysis (SMA) for investigating comprehensive upregulation and downregulation of differentially expressed genes (DEGs) among different stem cell populations. Materials and Methods: A total of 55 mouse and human data sets targeting crucial genes identified in seven different types of stem cells population were screened and subjected to independent DEGs analysis using SMA. Identified 30 meta-gene signatures were subjected to functional enrichment analysis based on their biological processes and molecular functions. Validation of enriched meta-gene signatures was performed using RT-qPCR. Cellular localization of ABCB1 and ABCG2 was identified using immunofluorescence staining, whereas functional assessment was performed using western-blot. Results: SMA analysis revealed that among 52 commonly expressed genes, 30 genes were either upregulated or downregulated in at least two stem cell populations. Further gene enrichment analysis showed nine genes (ABCB1, ABCG2, HSPA4, HSPA9, HSPA14, Nestin, Sox-2, Oct-4, and Notch-2) with the highest combined scores among 30 meta-gene signatures. RT-qPCR demonstrated that all the enriched gene signatures were significantly upregulated in primary NPCs and further downregulated during NPCs lineage differentiation in culture except HSPA4, HSPA9, and HSPA14 gene transcripts. Conclusions: The stemness meta-gene signatures were abundantly expressed in human NPCs population which categorically suggest the involvement of these genes/pathways in pluripotency maintenance and molecular switches for lineage differentiation while HSP-70 had a neuroprotective effect.

Fabrication of Decellularized Amnion and Chorion Scaffolds to Develop Bioengineered Cell-Laden Constructs.

INTRODUCTION: Human mesenchymal stem cells (hMSCs) holds great promise for managing several clinical conditions. However, the low engraftment efficiency and obscurity to harvest these cells without compromising the cellular viability, structural and functional properties from the culture niche still remain major obstacles for preparing intact regenerative constructs. Although few studies have demonstrate different methods for generating cell-liberated amniotic scaffolds, a common method for producing completely cell-liberated amnion (D-HAM) and chorion (D-HCM) scaffolds and their cytocompatibility with hMSCs yet to be demonstrated. METHODS: A common process was developed for preparing D-HAM and D-HCM scaffolds for assessing hMSCs engraftment efficiency, proliferation and molecular shifts to generate cell-laden biological discs. The structural and functional integrity of D-HAM and D-HCM was evaluated using different parameters. The compatibility and proliferation efficiency of hMSCs with D-HAM and D-HCM was evaluated. RESULTS: Histological analysis revealed completely nucleic acid-free D-HAM and D-HCM scaffolds with intact extracellular matrix, mechanical and biological properties almost similar to the native membranes. Human MSCs were able to adhere and engraft on D-HCM better than D-HAM and expanded faster. Ultrastructural observations, crystal violet staining and expression studies showed better structural and functional integrity of hMSCs on D-HCM than D-HAM and control conditions. CONCLUSION: A common, simple and reliable process of decellularization can generate large number of cell-liberated amniotic scaffolds in lesser time. D-HCM has better efficiency for hMSCs engraftment and proliferation and can be utilized for preparing suitable cell-laden constructs for tissue engineering applications.

Long-term and non-invasive in vivo tracking of DiD dye-labeled human hepatic progenitors in chronic liver disease models.

BACKGROUND: Chronic liver diseases (CLD) are the major public health burden due to the continuous increasing rate of global morbidity and mortality. The inherent limitations of organ transplantation have led to the development of stem cell-based therapy as a supportive and promising therapeutic option. However, identifying the fate of transplanted cells in vivo represents a crucial obstacle. AIM: To evaluate the potential applicability of DiD dye as a cell labeling agent for long-term, and non-invasive in vivo tracking of transplanted cells in the liver. METHODS: Magnetically sorted, epithelial cell adhesion molecule positive (1 × 106 cells/mL) fetal hepatic progenitor cells were labeled with DiD dye and transplanted into the livers of CLD-severe combined immunodeficiency (SCID) mice. Near-infrared (NIR) imaging was performed for in vivo tracking of the DiD-labeled transplanted cells along with colocalization of hepatic markers for up to 80 d. The existence of human cells within mouse livers was identified using Alu polymerase chain reaction and sequencing. RESULTS: NIR fluorescence imaging of CLD-SCID mice showed a positive fluorescence signal of DiD at days 7, 15, 30, 45, 60, and 80 post-transplantation. Furthermore, positive staining of cytokeratin, c-Met, and albumin colocalizing with DiD fluorescence clearly demonstrated that the fluorescent signal of hepatic markers emerged from the DiD-labeled transplanted cells. Recovery of liver function was also observed with serum levels of glutamic-oxaloacetic transaminase, glutamate-pyruvate transaminase, and bilirubin. The detection of human-specific Alu sequence from the transplanted mouse livers provided evidence for the survival of transplanted cells at day 80. CONCLUSION: DiD-labeling is promising for long-term and non-invasive in vivo cell tracking, and understanding the regenerative mechanisms incurred by the transplanted cells.

Antibiograms of Gut Flora of Poultry Farms Workers Reveal Higher Resistance Levels as Compared to Non-Workers.

Introduction: Antibiotics are being used in humans and animals for treatment and control of bacterial infections. Excessive use of antibiotics in the production of poultry is a popular practice, but it poses serious health issues by transferring resistance from farm to humans via food or direct exposure. Study Objective: The objective of this study was to carry out a comparison of the resistance and sensitivity profile of isolated isolates from sewage of toilets that were in use of workers inside the farm and from sewage of household toilets. Methodology: In this study, a total of 320 sewage samples were collected. The antibiotic susceptibility profile was checked by Kirby-Bauer disc diffusion method, and the statistical analysis was carried out by MS excel. Chi-square test was performed to determine whether the antibiograms from two sample types were statistically different from each other or not. Results: From 320 sewage samples, a total of 296 bacterial isolates were isolated among which the leading bacterium was E. coli. The proportion of resistance, ESBL production and MDR was significantly higher in bacteria isolated from sewage of toilets under use of poultry farm workers as compared to the sewage from domestic use toilets. Conclusion: Resistance significantly increased in the bacteria isolated from toilets under use of poultry farm workers as compared to the ones isolated from control sewage samples.