Real-time Reverse Transcription-polymerase Chain Reaction Research Articles

Seroprevalence and risk factors of Bluetongue virus in camels in Northern Oman: a cross-sectional study.

Bluetongue virus (BTV) has emerged as a significant concern in Oman, affecting various animal species, including camels. This cross-sectional study aimed to assess the seroprevalence of BTV in camels and explore the associated risk factors within the northern region of Oman. Between October 2016 and March 2017, 439 serum samples and 100 blood samples were collected from camels in five governorates. Competitive enzyme-linked immunosorbent assay (c-ELISA) was employed to screen the serum for BTV antibodies, and real-time reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect BTV genomic RNA in blood. The findings revealed an overall seroprevalence of BTV of 56.04% (246 out of 439), with a 95% confidence interval ranging from 51.3% to 60.7%. BTV seropositivity was significantly associated with various factors, including geographical location, camel age, sex, participation in events, and a history of previous outbreaks. However, no significant correlation was found with tick infestations on farms or a history of abortion. These results underscore the enzootic and widespread nature of BTV in northern Oman, with camels appearing to manifest BTV infection as an asymptomatic or subclinical condition. To gain a more comprehensive understanding, further investigations are essential to identify the specific BTV serotypes currently in circulation within the Sultanate of Oman. The implications of these findings extend to the development of targeted interventions to manage and control the spread of BTV in this region.

Comparison of Commercially Available Thermostable DNA Polymerases with Reverse Transcriptase Activity in Coupled Reverse Transcription Polymerase Chain Reaction Assays

Reverse transcription polymerase chain reaction (RT-PCR) is an important tool for the detection of target RNA molecules and the assay of RNA pathogens. Coupled RT-PCR is performed with an enzyme mixture containing a reverse transcriptase and a thermostable DNA polymerase. To date, several biotechnological companies offer artificial thermostable DNA polymerases with a built-in reverse transcriptase activity for use in the coupled RT-PCR instead of the enzyme mixtures. Here, we compared the artificial DNA polymerases and conventional enzyme mixtures for the RT-PCR by performing end-point and real-time RT-PCR assays using severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV2) RNA and endogenous mRNA molecules as templates. We found that the artificial enzymes were suitable for different RT-PCR applications, including SARS-CoV2 RNA detection but not for long-fragment RT-PCR amplification.

Liraglutide mitigates dexamethasone-induced fatty acid synthase (FASN) and the cluster of differentiation36 (CD36) expression: a potential treatment for glucocorticoid-induced non-alcoholic fatty liver disease (NAFLD).

The clinical use of dexamethasone (DXM) is associated with the development of non-alcoholic fatty liver disease (NAFLD). However, the mechanisms by which DXM-induced NAFLD is still incompletely known. Therefore, the current study aims to test the hypothesis that DXM-induced NAFLD is mediated by dysregulation of key genes involved in lipid metabolism and liraglutide (LG) can ameliorate these effects. The histopathological and biochemical analysis assessed the effects of DXM and/or LG in liver tissue. The computational analysis was performed to detect the glucocorticoid response elements (GRE) in the promotor regions of FASN and CD36 genes. The effects of DXM and LG on the expression of FASN and CD36 were determined by real-time quantitative reverse transcription PCR (RT-qPCR), western blot (WB) and immunohistochemical (IHC) analyses. The NAFLD induced by high-fat diet (HFD) and DXM was manifested by increased levels of liver enzymes, deterioration of histological architecture of the liver tissue and accumulation of fat droplets. Computational analysis revealed that the promotor regions of FASN and CD36 harper several GRE. Most importantly, treatment with DXM decreased phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK) levels, while LG upregulate it. In addition, treatment with DXM increased expression of FASN and CD36, whereas LG ameliorated these effects in a dose-dependent manner. DXM-induced NAFLD is mediated by upregulation of FASN and CD36 expression which may be attributed to GRE. LG coupling with DXM mitigates this induction by downregulating FASN and CD36 levels and therefore mitigates the development of NAFLD.

TSPOAP1-AS1: A Novel Biomarker for the Prognosis and Therapeutic Target in Cervical Cancer.

TSPOAP1 antisense RNA 1 (TSPOAP1-AS1) is a long non-coding RNA (lncRNA) that has received widespread attention in oncology research in recent years. Its role and mechanism in some cancers have gradually been revealed. However, it is not clear what role TSPOAP1-AS1 plays in cervical cancer (CESC). In this study, bioinformatic analysis and experimental validation were carried out to investigate the relationship between TSPOAP1-AS1 and CESC. The relationships between clinical characteristics in patients with CESC, TSPOAP1-AS1 expression, prognostic factors, regulation network, and immune infiltration of TSPOAP1-AS1 were evaluated using statistics and The Cancer Genome Atlas database. Real-Time Quantitative Reverse Transcription PCR was used to test TSPOAP1-AS1, miR-17-5p, and AGFG2 expression in CESC cell lines. CESC patients exhibited markedly reduced expression of TSPOAP1-AS1. There was a significant correlation between low expression of TSPOAP1-AS1 in CESC patients and the clinical stage (p < 0.05), weight (p < 0.05), and BMI (p < 0.05). Lower expression of TSPOAP1-AS1 in patients with CESC was associated with poorer overall survival (OS) (p = 0.014) and disease-specific survival (DSS) (p = 0.030). There was also an independent correlation between high expression of TSPOAP1- AS1 (p = 0.036) and DSS in patients with CESC. TSPOAP1-AS1 was involved in the ribosome, oxidative phosphorylation, antigen processing and presentation, cell adhesion molecules (CAMs), the chemokine signaling pathway, neuroactive ligand-receptor interaction, and primary immunodeficiencies. The infiltration of immune cells and the expression of TSPOAP1-AS1 were found to be correlated. A ceRNA network of TSPOAP1-AS1/miR-17-5p/AGFG2 was constructed in CESC. In CESC, a ceRNA network involving TSPOAP1-AS1/miR-17-5p/AGFG2 was successfully established. When comparing CESC cell lines with HcerEpic, the expression of TSPOAP1-AS1 and AGFG2 decreased significantly, and the expression of miR-17-5p increased significantly. In CESC patients, low expression of TSPOAP1-AS1 was associated with poor survival and immune infiltration. It may be effective to use TSPOAP1-AS1 as a biomarker of prognosis and therapeutic target in CESC.

Mir-615-5p inhibits cervical cancer progression by targeting TMIGD2

BackgroundCervical cancer (CC) is a prevalent gynecological malignancy, contributing to a substantial number of fatalities among women. MicroRNAs (miRNAs) have emerged as promising biomarkers with significant potential for the early detection and prognosis of CC.ObjectiveThis study aimed to explore the clinical significance and biological role of miR-615-5p in CC, with the goal of identifying novel biomarkers for this disease.Materials and methodsThe levels of miR-615-5p and TMIGD2 mRNA in tissue samples and cells were quantified through quantitative reverse transcription real-time PCR, followed by statistical analyses to investigate the correlation between miR-615-5p and clinical data. The effects of miR-615-5p on the proliferation and metastasis of CC cells were evaluated using the Cell Counting Kit-8 and Transwell assays. The potential mechanism of miR-615-5p was elucidated by bioinformatics analyses and Dual-luciferase reporter assay. Western blotting was employed to measure the protein levels of TMIGD2.ResultsIn CC, the downregulation of miR-615-5p was related to poor prognosis and emerged as an independent prognostic factor. The levels of miR-615-5p were reduced in CC cells. miR-615-5p overexpression restrained the proliferation and metastasis of CC cells. Furthermore, TMIGD2 was identified as a target gene regulated by miR-615-5p, and its expression was notably elevated in CC. The influence of miR-615-5p on the biological behaviors of CC cells was mediated through the modulation of TMIGD2.ConclusionsDownregulation of miR-615-5p was a prognostic indicator of poor prognosis in CC. miR-615-5p exerted its tumor-suppressive effects by inhibiting cell growth and metastasis through the regulation of TMIGD2.

Zeolite Imidazole Framework-8 Exacerbates Astrocyte Activation and Oxidative Stress in the Brain of Rats.

Metal-organic frameworks (MOFs) have been gaining significant attention due to their potential application in medicine. Here, we investigated the effect of zeolite imidazole framework-8 (ZIF-8) on neuro-behavioral parameters, histopathology, inflammation, and oxidative stress levels of rats' brain samples. Forty-eight male Wistar rats were injected by four injections of saline or ZIF-8 at different doses of 5, 10, or 20 mg/kg via the caudal vein. Y-Maze, Morris-Water Maze (MWM), and three chamber tests were conducted to explore working memory, spatial learning and memory, and social interactions, respectively. Histological staining and immunohistochemistry were used to evaluate pathological changes and astrocyte activation levels. The inflammation levels were measured using quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). The total antioxidant capacity (TAC) and oxidative stress production were assessed by biochemical assays. The results showed that ZIF-8 induces neuromotor impairment dose-dependently. Although histopathological studies indicated increased neuronal loss, inflammatory changes, and elevated active astrocytes in the hippocampus, the expression levels of IL-1β and TNF-α were not significantly increased in ZIF-8-treated rats. The TAC level significantly reduced and the malondialdehyde (MDA) level remarkably increased in the brain tissues. Our findings suggest that administration of ZIF-8 induce neuromotor impairment, probably through amplified inflammation and oxidative stress.

A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus.

To establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the M gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) reaction mixture was developed. A simple and rapid RT-qPCR detection method for PRRSV was developed by optimizing nucleic acid amplification conditions. The results showed that the method was able to specifically detect PRRSV without cross-reactivity with the other 11 porcine susceptible viruses. The sensitivities of the assay were 3.12 × 100 copies/μL and 100 TCID50/μL for M gene and virus, respectively, and the repeatability and reproducibility (relative standard deviation, CV) of the assay were less than 2.5%. Based on the new fullly pre-mixed RT-qPCR reaction mixture, the RT-qPCR detection method may provide a new, simple, and rapid method for accurately detecting PRRSV.

Molecular diagnosis and phylogenetic analysis of a Middle East respiratory syndrome coronavirus human case in Jordan.

Middle East respiratory syndrome coronavirus (MERS-CoV) is an important zoonotic pathogen. The aim of this paper is to report one polymerase chain reaction (PCR)-positive case of MERS-CoV in a 27-year-old man who was involved in a nationwide longitudinal surveillance study of certain zoonotic diseases in Jordan including MERS-CoV. Whole-blood and nasal swab samples were collected from the man and five camels in the vicinity of his living area. The samples were subjected to enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription PCR (RT-PCR) to detect MERS-CoV-specific antibodies and MERS-CoV genetic material, respectively. Genomic sequencing and phylogenetic analysis were also performed to detect similarities with known strains of the virus in the region. In January 2021, an ongoing surveillance study detected a MERS-CoV-positive nasal swab sample from an asymptomatic male and camels using RT-PCR. Phylogenetically, the MERS-CoV isolated in this case belonged to clade B and is clustered with other strains originating in the Arabian Peninsula. The case report represents the first PCR-positive case of MERS-CoV in an asymptomatic individual in Jordan, indicating active circulation of the virus within the population.

Temporal Trends in Respiratory Infection Epidemics Among Pediatric Inpatients Throughout the Course of the COVID-19 Pandemic From 2018 to 2023 in Fukushima Prefecture, Japan.

Nonpharmaceutical interventions for coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2, during the pandemic altered the epidemiology of respiratory viruses. This study aimed to determine the changes in respiratory viruses among children hospitalized from 2018 to 2023. Nasopharyngeal specimens were collected from children aged under 15 years with fever and/or respiratory symptoms admitted to a medical institution in Fukushima Prefecture between January 2018 and December 2023. Eighteen respiratory viruses were detected using real-time reverse transcription-polymerase chain reaction. Overall, 1933 patients were included. Viruses were detected in 1377 (71.2%); of these, a single virus was detected in 906 (46.9%) and multiple viruses in 471 (24.3%). Among the viruses whose epidemics were temporarily suppressed, the epidemics of respiratory syncytial virus A and human parainfluenza virus type 3 (HPIV3) started earlier, and the epidemics of human metapneumovirus, HPIV1, and influenza A and C viruses resumed as behavioral restrictions for preventing COVID-19 eased. The median age of children with airway infection was significantly higher in the postpandemic group than in the prepandemic group (18.0 months vs. 21.0 months, p < 0.01). The median age of children infected with HPIV3 and human rhinovirus was significantly higher in the postpandemic group than in the prepandemic group. Strengthening of nonpharmaceutical interventions changed the epidemic dynamics of pediatric infectious diseases, with a trend toward older hospitalized children. Continuous monitoring of pediatric infectious disease outbreaks in hospitalized children can help prepare for the emergence of future viruses and pandemics.

The Blood-Cerebrospinal Fluid Barrier as a Potential Entry Site for the SARS-CoV-2 Virus.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is an RNA virus responsible for coronavirus disease 2019 (COVID-19). While SARS-CoV-2 primarily targets the lungs and airways, it can also infect other organs, including the central nervous system (CNS). The aim of this study was to investigate whether the choroid plexus could serve as a potential entry site for SARS-CoV-2 into the brain. Tissue samples from 24 deceased COVID-19-positive individuals were analyzed. Reverse transcription real-time PCR (RT-qPCR) was performed on selected brain regions, including the choroid plexus, to detect SARS-CoV-2 viral RNA. Additionally, immunofluorescence staining and confocal microscopy were used to detect and localize two characteristic proteins of SARS-CoV-2: the spike protein S1 and the nucleocapsid protein. RT-qPCR analysis confirmed the presence of SARS-CoV-2 viral RNA in the choroid plexus. Immunohistochemical staining revealed viral particles localized in the epithelial cells of the choroid plexus, with the spike protein S1 detected in the late endosomes. Our findings suggest that the blood-cerebrospinal fluid (B-CSF) barrier in the choroid plexus serves as a route of entry for SARS-CoV-2 into the CNS. This study contributes to the understanding of the mechanisms underlying CNS involvement in COVID-19 and highlights the importance of further research to explore potential therapeutic strategies targeting this entry pathway.

The Dynamic Changes of COL11A1 Expression During the Carcinogenesis and Development of Breast Cancer and as a Candidate Diagnostic and Prognostic Marker.

Purpose: Collagen type XI alpha 1 (COL11A1), a critical member of the collagen superfamily, is essential for tissue structure and integrity. This study aimed to validate previously identified variations in COL11A1 expression during breast cancer carcinogenesis and progression, as well as elucidate their clinical implications. Methods: COL11A1 mRNA expression levels were assessed using real-time reverse transcription-PCR (RT-PCR) in 30 pairs of normal breast tissue and primary breast cancer, 30 pairs of primary breast cancer and lymph node metastases, 30 benign tumors, and 107 primary breast cancers. COL11A1 protein expression was evaluated by Western blot in six matched trios of normal tissue, primary cancer, and lymph node metastasis. Results: COL11A1 mRNA levels were significantly higher in primary breast cancer tissues (n = 30) than in adjacent normal breast tissues (p < 0.001). Conversely, lymph node metastases (n = 30) showed significantly lower COL11A1 mRNA levels compared to their primary breast cancer counterparts (p=0.005). In a larger cohort, primary breast cancers (n = 107) had significantly elevated COL11A1 mRNA levels relative to adjacent normal tissues (n = 30) and benign tumors (n = 30) (p < 0.001). Benign tumors also demonstrated higher levels compared to normal tissues (p=0.012). The protein expression patterns were consistent with the mRNA findings. Receiver operating characteristic (ROC) curve analysis confirmed the diagnostic relevance of COL11A1 expression levels. Significant associations were found between COL11A1 mRNA levels and clinical parameters including lymph node involvement (p=0.046), clinical stage (p=0.004), and progesterone receptor status (p=0.048). Overexpression of COL11A1 was correlated with poor prognosis. Conclusions: COL11A1 expression varies during breast tumor initiation and progression, with elevated levels linked to worse prognoses. These findings underscore COL11A1's potential as a biomarker in breast cancer, suggesting its assessment could enhance diagnostic and prognostic strategies for more personalized patient management.

Exploring the Analgesic Effect of Acupuncture on Knee Osteoarthritis Based on MLT/cAMP/PKA/CREB Signaling Pathway.

Acupuncture is an effective treatment for knee osteoarthritis (KOA), reducing pain and improving function. While melatonin (MLT) has notable pain relief benefits, the analgesic mechanism of acupuncture in KOA and its relationship with melatonin are still unknown. This study aims to explore this mechanism. In this work, the KOA rabbit model was constructed using the traditional Hulth method, and the therapeutic effect was assessed by the Lequesne MG score and Pain assessment by hot plate test. The pathological alterations of cartilage tissue were observed using hematoxylin and eosin (H&E) staining, Safranin O-fast green and MASSON staining to observe the pathological changes in cartilage tissue, and the efficacy was evaluated according to the principles of Mankin score and Osteoarthritis Research Society International (OARSI) score. Meanwhile, MLT in serum, cyclic adenosine monophosphate (cAMP) in cartilage, and matrix metalloproteinase-3 (MMP-3) in joint fluid were detected by enzyme-linked immunosorbent assay. In addition, the expression of aromatic L-amino acid N-acetyltransferase (AANAT), melatonin receptor 1 (MT1) and 2 (MT2) mRNAs in cartilage was determined by real-time quantitative reverse transcription-polymerase chain reaction, and the levels of proteins related to PKA/CREB signaling pathway were detected by Western blotting. Based on the results of Lequesne MG score and Pain assessment by hot plate test experimental data, the treatment group presented significant improvements in knee pain and overall function relative to OA (Osteoarthritis) group. Besides, according to results of histologic staining, Mankin and OARSI scores, articular cartilage degeneration of treatment group remarkably improved. In addition, acupuncture significantly reduced the expression of the inflammatory factor MMP-3 in knee joint fluid and significantly increased the levels of MLT, AANAT, MT1, MT2, cAMP, PKA and CREB. By regulating sympathetic excitability, acupuncture may activate the MLT/cAMP/PKA/CREB signaling pathway, decrease inflammatory factor expression and slow down degradation of articular cartilage, resulting in the relief of knee pain.

LINC02987 suppression hepatocellular carcinoma progression by modulating autophagy via the miR-338-3p/ATG12 axis.

Hepatocellular carcinoma (HCC), the most common primary liver cancer, is marked by a high mortality rate, with the misregulation of long non-coding RNAs (LncRNAs) playing a key role in its development. Here, we studied the role of LINC02987 in HCC. We employed bioinformatics tools to identify LncRNAs and miRNAs that exhibit differential expression in HCC. Quantitative real-time reverse transcription PCR (RT-qPCR) and Western blot analysis were utilized to quantify gene and protein expression levels. The interaction between miR-338-3p and LINC02987 or ATG12 was confirmed through dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. We observed that LINC02987 was overexpressed in HCC tumor tissues and cell lines. Silencing of LINC02987 led to a reduction in cell viability, diminished clonogenic potential, and attenuated invasive and migratory capabilities. Also, decreasing protein level and fluorescence intensity of the autophagy-associated LC3 I/II. In HCC, miR-338-3p expression was downregulated, while inversely correlates with the overexpression of the autophagy protein ATG12. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in corresponding reporter assays. Mimicking miR-338-3p suppresses the activity of both LINC02987 and ATG12, as evidenced by reduced luciferase signals in reporter assays. Transfection with si-LINC02987 decreased ATG12 expression, an effect that was partially reversed by miR-338-3p knockdown. Inhibition of miR-338-3p or overexpression of ATG12 increased LC3 I/II protein levels. Our results indicate that LINC02987 sequesters miR-338-3p, leading to increased ATG12 and promoting autophagy in HCC cells. These results highlight the potential of LINC02987 as a therapeutic target for the treatment of HCC.

Predictive biosignatures for hospitalization in patients with virologically confirmed COVID-19.

COVID-19, caused by the SARS-CoV-2 virus, presents with varying severity among individuals. Both viral and host factors can influence the severity of acute and chronic COVID-19, with chronic COVID-19 commonly referred to as long COVID. SARS-CoV-2 infection can be properly diagnosed by performing real-time reverse transcription PCR analysis of nasal swab samples. Pulse oximetry, chest X-ray, and complete blood count (CBC) analysis can be used to assess the condition of the patient to ensure that the appropriate medical care is delivered. This study aimed to develop biosignatures that can be used to distinguish between patients who are likely to develop severe disease and require hospitalization from patients who can be safely monitored in less intensive settings. A retrospective investigation was conducted on 7897 adult patients with virologically confirmed SARS-CoV-2 infection between January 26, 2020, and November 30, 2023; all patients underwent comprehensive CBC testing at Taipei Veterans General Hospital). Among them, 1867 patients were independently recruited for a population study involving genome-wide genotyping of approximately 424 000 genomic variants. Therefore, the participants were divided into two patient cohorts, one with genomic data (n = 1867) and one without (n = 6030) for model validation and training, respectively. We constructed and validated a biosignature model by using a combination of CBC measurements to predict subsequent hospitalization events (hazard ratio [95% confidence interval] = 3.38, [3.07, 3.73] for the training cohort and 3.03 [2.46, 3.73] for the validation cohort; both p < 10-8). The obtained scores were used to identify the top quartile of patients, who formed the "very high risk" group with a significantly higher cumulative incidence of hospitalization (log-rank p < 10-8 in both the training and validation cohorts). The "very high risk" group exhibited a cumulative hospitalization rate of >60%, whereas the rate for the other patients was approximately 30% over a 1.5-year period, providing a binary classification of patients with distinct hospitalization risks. To investigate the genetic factors mediating this risk, we conducted a genome-wide association study. Specific regions in chromosomes 7 and 10 and the mitochondrial chromosome (M), harboring IKZF1, ABLIM1 and MT-ND3, exhibited prominent associations with binary risk classification. The identified exonic variants of IKZF1 are linked to several autoimmune diseases. Notably, people with different genotypes of the leading variants (rs4132601, rs141492519, and Affx-120744614) exhibited varying cumulative hospitalization rates following infection. We successfully developed and validated a biosignature model of COVID-19 severe disease in virologically confirmed patients. The identified genomic variants provide new insights for infectious disease research and medical care.

CircTLK1 Knockdown Alleviates Cell Inflammation and Apoptosis by Regulating Elavl1/Nox4 Axis in Neonatal Sepsis-Induced Lung Injury in Mice.

Septic acute lung injury (ALI) is a common complication of sepsis with high morbidity and mortality but lacks specific treatment. This study aimed to elucidate the role of circular RNA TLK1 (circTLK1) in neonatal septic ALI. Murine cecal slurry was used to induce neonatal sepsis-induced ALI model in vivo. Hematoxylin and eosin staining was performed to detect the pathological changes in lung tissues. Pulmonary microvascular endothelial cells were treated with lipopolysaccharide (LPS) to induce neonatal sepsis-induced ALI model in vitro. The levels of IL-1β and IL-6 were detected by enzyme-linked immunosorbent assay. A lactate dehydrogenase (LDH) detection kit was used to detect LDH activity. Cell Counting Kit-8 assay and flow cytometry detected cell viability and apoptosis. The genes' expression was measured by quantitative real-time reverse-transcription polymerase chain reaction and western blot. The relationship between circTLK1 and Elavl1 or Elavl1 and Nox4 was detected using RNA immunoprecipitation assay. Our results illustrated that circTLK1 was highly expressed in neonatal sepsis-induced ALI model, and circTLK1 knockdown alleviated cell inflammation and apoptosis in neonatal sepsis-induced ALI model. Similarly, we found that circTLK1 knockdown alleviated neonatal sepsis-induced ALI. Mechanically, circTLK1 mediated Elavl1 binding to Nox4 messenger RNA and increased its stability. Functionally, circTLK1 knockdown alleviated cell inflammation and apoptosis by regulating Nox4 in the neonatal sepsis-induced ALI model. CircTLK1 knockdown alleviated cell inflammation and apoptosis by the Elavl1/Nox4 axis in neonatal sepsis-induced ALI. Our research provided a novel direction for the treatment of neonatal sepsis-induced ALI. · CircTLK1 knockdown relieved neonatal septic ALI.. · CircTLK1 mediated Elavl1 binding to Nox4 mRNA..

The transcription factor YbdO attenuates the pathogenicity of avian pathogenic Escherichia coli by regulating oxidative stress response

Avian pathogenic Escherichia coli (APEC) is a significant pathogen infecting poultry that is responsible for high mortality, morbidity and severe economic losses to the poultry industry globally, posing a substantial risk to the health of poultry. APEC encounters reactive oxygen species (ROS) during the infection process and thus has evolved antioxidant defense mechanisms to protect against oxidative damage. The imbalance of ROS production and antioxidant defenses is known as oxidative stress, which results in oxidative damage to proteins, lipids and DNA, and even bacterial cell death. APEC uses transcription factors (TFs) to handle oxidative stress. While many TFs in E. coli have been well characterized, the mechanism of the YbdO TF on protecting against oxidative damage and regulating the virulence and pathogenicity of APEC has not been clarified. Here we focus on the regulatory mechanism of YbdO on the pathogenicity of APEC. The results from this study showed that YbdO attenuated the pathogenicity of APEC in chicks infection models by inhibiting the expression of virulence genes fepG and ycgV using quantitative real-time reverse transcription PCR (RT-qPCR) experiments. The electrophoretic mobility shift assays (EMSA) confirmed that YbdO specifically bound to the promoters of fepG and ycgV. Additionally, YbdO increases H2O2-induced oxidative damage to APEC via repressing the expression of oxidative stress response genes sodA, soxR, ahpC, ahpF, katG, and oxyR by binding to their promoter regions. The repression effect facilitates host immune response to eliminate APEC and to generate beneficial immune protection to the body, thereby indirectly attenuating the pathogenicity of APEC. These findings might provide further insights into the mechanism of oxidative damage to APEC and offer new perspectives for further studies on the prevention and control of APEC infections.

Canid alphaherpesvirus 1 infection alters the gene expression and secretome profile of canine adipose-derived mesenchymal stem cells in vitro

BackgroundCanine adipose-derived mesenchymal stem cells (cAD-MSCs) demonstrate promising tissue repair and regeneration capabilities. However, the procurement and preservation of these cells or their secreted factors for therapeutic applications pose a risk of viral contamination, and the consequences for cAD-MSCs remain unexplored. Consequently, this research sought to assess the impact of canid alphaherpesvirus 1 (CHV) on the functional attributes of cAD-MSCs, including gene expression profiles and secretome composition.MethodsTo this end, abdominal adipose tissue from 12 healthy dogs was harvested to isolate cAD-MSCs. These samples were tested for CHV contamination before introducing a wild-type CHV strain via serial passages. Following CHV infection, real-time reverse transcription-polymerase chain reaction array and liquid chromatography with tandem mass spectrometry assessments enabled analyses of gene expression and secretome’s proteomic profile, respectively.ResultsThis study showed that the initial cAD-MSC populations were devoid of CHV. cAD-MSCs showed susceptibility to infection with wild-type CHV, leading to notable modifications in gene expression and secretome profile. The observed genomic variations in gene expression indicate potential impacts on the stemness, migration, and other functional properties of cAD-MSCs, highlighting the need for further studies to evaluate their functional capacity post-infection. Moreover, gene expression and secretome analyses suggest a shift in stem cell differentiation toward an adipogenic phenotype.ConclusionTo the best of our knowledge, this is the first study of the effects of virus infection on gene expression and secretome composition in cAD-MSCs. The outcomes of our study underscore the imperative of routine viral screening prior to the therapeutic use of cAD-MSCs. Moreover, these findings provide novel insights into the pathogenic mechanisms of CHV and pave the way for future canine stem cell and virus research.

A Nationwide Population-based Study for Audio-vestibular Disorders Following COVID-19 Infection.

To date, no study has reported the various otologic conditions associated with COVID-19 using population-based design. The aim of this study was to investigate the incidence and risk of audio-vestibular disorders (benign paroxysmal positional vertigo, sudden sensorineural hearing loss, Meniere's disease, vestibular neuritis, and tinnitus) increase after COVID-19 infection. This retrospective population-based study was conducted using National Health Insurance Service (NHIS)-COVID-19 cohort database of South Korea. We identified participants in the COVID-19 group using real-time reverse transcription-polymerase chain reaction tests. A matched cohort without COVID-19 was randomly selected in a 1:1 ratio. Benign paroxysmal positional vertigo, sudden sensorineural hearing loss, Meniere's disease, vestibular neuritis, and tinnitus were defined using diagnostic, medication, and procedure codes. The incidence and risk of these disorders were assessed in both groups using univariate and multivariate Cox proportional hazard analyses. In total, 4,976,589 COVID-19 patients and an equivalent number of matched non-infected controls were analyzed. COVID-19 patients faced an increased risk of developing benign paroxysmal positional vertigo, sudden sensorineural hearing loss, vestibular neuritis, and tinnitus compared to controls in univariate and multivariate Cox hazard analyses. COVID patients were at an increased risk of Meniere's disease in the univariate analysis; however, the risk of Meniere's disease after COVID-19 did not reach statistical significance in the multivariate analysis. COVID-19 infection may increase the risk of benign paroxysmal positional vertigo, sudden sensorineural hearing loss, vestibular neuritis, and tinnitus.

The absence of the Bluetongue virus in abortion cases in cattle, sheep, and goats in Türkiye: 2012-2017

Bluetongue (BT) is a disease that affects domestic and wild ruminants, and it is caused by a virus called bluetongue virus (BTV) that is transmitted by Culicoides midges. Although clinical signs of BT are most apparent in sheep, BTV could induce abortion and birth defects in cattle, sheep and goats. BTV infection has been reported in Türkiye, but the role of BTV in cattle and small ruminant abortion cases in Türkiye remains uncertain. Therefore, this research aimed to fill this research gap by investigating the prevalence of BTV in cattle and small ruminant abortion cases. To investigate the frequency of BTV in ovine, caprine, and bovine foetuses, a total of 1718 foetuses were collected from different farms between 2012 and 2017. A one-step real-time reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect BTV RNA in aborted foetuses. BTV specific RNA was not detected in the analysed foetuses. To the best of my knowledge, this is the longest study that has investigated whether BTV infection has a role in cattle and small ruminant abortion cases in Türkiye. The results of this study are limited only to the regions studied. Therefore, further epidemiological studies are needed to confirm the findings of this study.

Detection of Sars-Cov-2 Nucleic Acid by Rt-Pcr in Symptomatic and Asymptomatic Patients in Dhaka, Bangladesh

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the underlying etiology of COVID-19, a contagious respiratory disease. Proper diagnosis is required to prevent infection from spreading among persons and communities. Precise viral detection is a prerequisite to control the COVID-19 pandemic. Thus, the objective of this study is the detection of SARS-CoV-2 nucleic acid by Real-Time Reverse Transcription-Polymerase Chain Reaction (Real-Time RT-PCR) method of the suspected symptomatic and asymptomatic individuals in Dhaka, Bangladesh. This research was performed in different local hospitals in Dhaka, Bangladesh from June 2021 to January 2022. Samples from suspected individuals were collected following WHO guidelines. Then the samples were placed into a Viral Transport Medium (3 ml sterile VTM). After vortexing, Sansure Biotech Inc. virus nucleic acid RT-PCR test kits were used for diagnosis. Real-Time RT-PCR method was used to identify the positive and negative results of the samples. A total number of 416 COVID-19-suspected individuals were enrolled in this study; 297 (71%) tested real-time RT-PCR-positive, and 119 (29%) were negative. The test results of patients with COVID-19 symptoms were collected and analyzed. Among the positive samples, the positive rate of male patients was higher than female patients. Between age groups, 20 – 59 has shown the highest positivity rate and the lowest was observed below age group 10. Age distribution and clinical characteristics were compared with the neighboring country’s COVID-19 outbreak survey and found to be quite similar. Real-time RT-PCR for SARS-CoV-2 remains the gold standard identification method to verify COVID-19 cases unless and until the World Health Organization (WHO) recommends any alternative approach. Early detection at the primary level of SARS-CoV-2 may aid the clinicians in treatment of COVID-19. All the necessary guidelines need to be followed even after the post-COVID-19 situation. Stam. J. Microbiol. 2024;14(1):13-17