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Introduction: Retinoblastoma (RB) is the most common paediatric intraocular malignancy. The Spleen Tyrosine Kinase (SYK) acts as a proto-oncogene crucial for RB cell survival and is absent in the normal retina, making it a valuable therapeutic target. Aim: Entospletinib, a selective oral SYK inhibitor currently in phase 2 trials for chronic lymphocytic leukaemia, was evaluated for its activity on the RB cell line Y-79. Materials and Methods: The effects of Entospletinib on cell proliferation and death of the human RB Y79 cells were assessed by MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay, phase contrast microscopy and AO/EtBr dual labeling. The Annexin-V kit was used to quantify the cell mortality following treatment using flow cytometry. The mRNA levels of caspase-3, BAX, BCL-2, SYK, p53, TNF-and NF-κB were measured using RT-PCR to better understand the mechanism of action of this drug. Western blot was used to further validate the data. Results: Entospletinib treatment led to a time- and dosedependent reduction in cell viability, with significant cytotoxicity observed at an IC50 of 61.3 μM/ml. Annexin V/Propidium Iodide (PI) staining demonstrated a dose-dependent increase in apoptosis. The treatment upregulated mRNA levels of Caspase-3, Bax, and p53 while downregulating SYK, TNF-α, and Bcl-2. RT-PCR and Western blot analyses confirmed a dose-dependent decrease in NF-κB/p65 expression. Conclusion: The study findings suggest that Entospletinib inhibits cell proliferation and induces apoptosis in RB cells by downregulating SYK/TNF-α/NF-κB/p65, highlighting its potential as a promising therapeutic candidate for RB. [ABSTRACT FROM AUTHOR]

COVID-19 is a highly transmissible illness caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The World Health Organization (WHO) declared it a pandemic in 2020, following its rapid global spread. While the virus can lead to severe illness, particularly among vulnerable groups such as the elderly, individuals with compromised immune systems, diabetics, those with heart conditions, and patients with hypertension, many inf ected individuals remain asymptomatic or exhibit mild to moderate respiratory symptoms. The most critical manifestation of COVID-19 is often marked by a cytokine storm ( A cytokine storm : is an intense immune response marked by the excessive release of pro-inflammatory cytokines into the bloodstream. This can cause widespread inflammation and tissue damage across multiple organs. Commonly linked to severe infections, autoimmune diseases, and conditions like COVID-19, symptoms may include fever, fatigue, and respiratory distress. If untreated, it can lead to serious complications or death. Management typically focuses on reducing inflammation and moderating the immune response.) The definitive laboratory diagnosis relies on the identification of viral ribonucleic acid (RNA) through real-time polymerase chain reaction (RT-PCR) testing of samples obtained from nasal and oropharyngeal swabs. This testing method is most effective when conducted in the initial days following symptom onset. Serological tests play a crucial role in assessing the immune response, as both IgM and IgG antibodies can be detected approximately seven days after the onset of symptoms and may persist for over 200 days. However, the presence of antibodies does not necessarily indicate that an individ ual is no longer infectious, as this depends on their viral load and clinical status. Utilizing specific laboratory markers judiciously is essential, considering the disease's progression. Proper interpretation of these markers can enhance patient management, facilitate the identification of asymptomatic carriers, and assist in monitoring those with mild symptoms. [ABSTRACT FROM AUTHOR]

Introduction: Retinoblastoma (RB) is the most common paediatric intraocular malignancy. The Spleen Tyrosine Kinase (SYK) acts as a proto-oncogene crucial for RB cell survival and is absent in the normal retina, making it a valuable therapeutic target. Aim: Entospletinib, a selective oral SYK inhibitor currently in phase 2 trials for chronic lymphocytic leukaemia, was evaluated for its activity on the RB cell line Y-79. Materials and Methods: The effects of Entospletinib on cell proliferation and death of the human RB Y79 cells were assessed by MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay, phase contrast microscopy and AO/EtBr dual labeling. The Annexin-V kit was used to quantify the cell mortality following treatment using flow cytometry. The mRNA levels of caspase-3, BAX, BCL-2, SYK, p53, TNF-and NF-κB were measured using RT-PCR to better understand the mechanism of action of this drug. Western blot was used to further validate the data. Results: Entospletinib treatment led to a time- and dose-dependent reduction in cell viability, with significant cytotoxicity observed at an IC50 of 61.3 μM/ml. Annexin V/Propidium Iodide (PI) staining demonstrated a dose-dependent increase in apoptosis. The treatment upregulated mRNA levels of Caspase-3, Bax, and p53 while downregulating SYK, TNF-α, and Bcl-2. RT-PCR and Western blot analyses confirmed a dose-dependent decrease in NF-κB/p65 expression. Conclusion: The study findings suggest that Entospletinib inhibits cell proliferation and induces apoptosis in RB cells by downregulating SYK/TNF-α/NF-κB/p65, highlighting its potential as a promising therapeutic candidate for RB.

Canine distemper, a highly fatal systemic disease in domestic dogs and wild carnivores, has the second highest mortality rate after rabies and is responsible for a large number of animal deaths around the world. It is considered a major pathogen in the canine infectious respiratory disease complex. This paper reports the finding of a study conducted to detect and characterise canine distemper virus (Canine Morbillivirus) based on N gene. A total of 59 samples collected from cases of respiratory infections in dogs were subjected to N gene based reverse transcriptase polymerase chain reaction (RT-PCR) and eleven of them (18.64 per cent) were found positive. Sequencing and phylogenetic analysis revealed that all the canine distemper viruses obtained in the present study were related to Indian strains that were previously reported. However, the viruses from the same district were similar among themselves.

Background and Aim: Feline infectious peritonitis (FIP) is an infectious, immune-mediated, and fatal disease in cats caused by a mutant feline coronavirus (FCoV) infection. Feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) are two common retroviruses that play a role in reducing feline immune function with opportunistic retrovirus infection being a predisposing factor for the development of FIP. This study aimed to evaluate the clinicopathological parameters of FIP in cats with and without retrovirus coinfection. Materials and Methods: In total, 62 cats presenting with pleural and/or peritoneal effusion at the Kasetsart University Veterinary Teaching Hospital, Bangkok, Thailand, were selected for the study. Effusion samples were collected and a reverse transcriptase-polymerase chain reaction (RT-PCR) assay was performed on all samples using the 3' untranslated region primer. All FCoV-positive cats were tested for retrovirus infection using a commercial kit (Witness FeLV-FIV [Zoetis]; United States). Clinical signs, hematological, and biochemical parameters of these cats were investigated and grouped. Results: Of the 62 cats with pleural and/or peritoneal effusion, FCoV was detected in 32, of which 21 were highly suspicious for FIP. The cats suspected of FIP were divided into three subgroups following viral detection. A total of 14 had only FCoV infection (Group A), four had FCoV and FeLV infection (Group B), and three had FCoV, FeLV, and FIV infection (Group C). Of the rest, 11 had definitive diagnoses, which included three being FCoV and FeLV-positive (Group D), and eight were retrovirus-negative (Group E). Mild anemia and lymphopenia were found in cats infected with these three viruses. An albumin-to-globulin ratio lower than 0.5 was found in FIP cats with only FCoV infection. Conclusion: Typically, cats with clinical effusion and FIP, with and without retrovirus coinfection, had similar hematological findings. Clinical signs, blood parameters, fluid analysis with cytological assessment, and RT-PCR assays could identify better criteria to diagnose FIP with and without retrovirus coinfection.

Background and Aim: The coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) that attacks the respiratory and digestive tract. The SARS-CoV-2 showed systemic characteristics with various clinical symptoms from subclinical to fatal (causing death). Transmission of SARS-CoV-2 has been reported to occur from humans to pets (cats, dogs, tigers, ferrets, and poultry). Knowledge about the role of domestic animals in the transmission of SARS-CoV-2 to humans, and as reservoirs of this virus needs to be investigated further. This study aimed to detect the presence of SARS-CoV-2 in domestic animals such as dogs, cats, pigs, cows, birds, and bats that are often in contact with humans. Materials and Methods: A total of 157 samples, which included nasopharyngeal and oropharyngeal swabs, along with sera samples from domestic animals such as cats, pigs, cows, birds, and bats, were taken from Veterinary Hospitals, Veterinary Clinics, and farms around the Yogyakarta region. Detection of the virus was done using rapid detection of viral antigens, antibodies, and reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Results: The results showed that 5/157 (3.1%) samples found positive against the COVID-19 virus using a rapid antibody test; however, the results were negative on the rapid antigen and RT-PCR tests. Antibody-positive samples came from animals that had a history of household COVID-19 human infection. Conclusion: Thus, findings of the present study conclude that there is a potential for transmission of the COVID-19 virus between animals and humans.

Background: COVID-19 pandemic continues to be a public health threat. Rapid antigen tests (RATs) for the detection of SARS-CoV-2 infection will help in formulation of clinical and public health strategies for the control of transmission. In India, 49% of COVID-19 tests done are RATs.1 The sensitivity of RAT is 50.6–84%. RAT has specificity ranging from 99.3% to 100%.2 The present study aims at evaluating the RAT screening test done on people attending fever clinic of Shimoga Institute of Medical Sciences, Shivamogga. Aims and Objectives: The objectives of the study were to evaluate the COVID-19 RAT screening test with reverse transcriptase-polymerase chain reaction (RT-PCR) as the gold standard test done at fever clinic of SIMS, Shimoga. Materials and Methods: An observational, cross-sectional analytical study was conducted for a period of 1 month, May 2021. The study participants included all the people attending fever clinic of SIMS, Shimoga. Assuming the sensitivity of RAT to be 85%, power of 80%, and precision of 5%, the calculated sample size was 204. Considering non-response rate of 10%, the final sample size was 224. Secondary data regarding contact number of the people attending fever clinic were collected from the COVID-19 test register. Oral consent was taken after explaining about study and assuring confidentiality. Telephonic interview was done to collect relevant information. Analysis was done using Epi Info software version 7.2.4.0. Descriptive statistics such as percentages and analytical statistics such as Student’s t-test and Chi-square test were used. Results: Overall positivity rate was 43.7%. About 71% of people had contact history. Sensitivity and specificity of RAT test were found to be 64.2% and 97.2%, respectively, and were comparable with the previous studies. Significant difference was found (P

Canine distemper, a highly fatal systemic disease in domestic dogs and wild carnivores, has the second highest mortality rate after rabies and is responsible for a large number of animal deaths around the world. It is considered a major pathogen in the canine infectious respiratory disease complex. This paper reports the finding of a study conducted to detect and characterise canine distemper virus (Canine Morbillivirus) based on N gene. A total of 59 samples collected from cases of respiratory infections in dogs were subjected to N gene based reverse transcriptase polymerase chain reaction (RT-PCR) and eleven of them (18.64 per cent) were found positive. Sequencing and phylogenetic analysis revealed that all the canine distemper viruses obtained in the present study were related to Indian strains that were previously reported. However, the viruses from the same district were similar among themselves. [ABSTRACT FROM AUTHOR]

Background and Aim: The coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) that attacks the respiratory and digestive tract. The SARS-CoV-2 showed systemic characteristics with various clinical symptoms from subclinical to fatal (causing death). Transmission of SARS-CoV-2 has been reported to occur from humans to pets (cats, dogs, tigers, ferrets, and poultry). Knowledge about the role of domestic animals in the transmission of SARS-CoV-2 to humans, and as reservoirs of this virus needs to be investigated further. This study aimed to detect the presence of SARS-CoV-2 in domestic animals such as dogs, cats, pigs, cows, birds, and bats that are often in contact with humans. Materials and Methods: A total of 157 samples, which included nasopharyngeal and oropharyngeal swabs, along with sera samples from domestic animals such as cats, pigs, cows, birds, and bats, were taken from Veterinary Hospitals, Veterinary Clinics, and farms around the Yogyakarta region. Detection of the virus was done using rapid detection of viral antigens, antibodies, and reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Results: The results showed that 5/157 (3.1%) samples found positive against the COVID-19 virus using a rapid antibody test; however, the results were negative on the rapid antigen and RT-PCR tests. Antibody-positive samples came from animals that had a history of household COVID-19 human infection. Conclusion: Thus, findings of the present study conclude that there is a potential for transmission of the COVID-19 virus between animals and humans. [ABSTRACT FROM AUTHOR]

Introduction: Real time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) test, the gold standard test for Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) detection, is a tedious process and requires proficient workforce. Accurate and fast test results may permit more efficient use of protective and isolation resources and allow rapid therapeutic interventions. Aim: To evaluate the analytical performance characteristics of the Cepheid Xpert Xpress SARS-CoV-2 test, a rapid, automated molecular test for SARS-CoV-2 with gold standard RT-PCR test. Materials and Methods: This retrospective cohort study was conducted in Virus Research and Diagnostic Laboratory (VRDL) in Department of Microbiology at GGS Medical College, Faridkot from January 2021-June 2021. A total of 100 nasopharyngeal samples, collected from clinically suspected Coronavirus Diseae-2019 (COVID-19) cases admitted at GGSMC during 1st January-30th June 2021 were tested both by Xpert assay and RT-PCR test simultaneously, taking RT-PCR as the gold standard test. The data was analysed by MedCalc® statistical software version 19.6.4., and sensitivity, specificity, predictive values, likelihood ratios and the agreement between the two tests were calculated. Results: The mean age of the study participants was 46 years. Of these, 55 were males and 45 were females. The overall sample sensitivity and specificity of the Xpert assay were both 100% and there was perfect agreement across specimens, if authors, set a cut-off Cycle threshold-value (Ct-value) at 40 cycles for Xpert. Of 100 samples, 32 were positive for SARSCoV-2 by either of the tests and 68 were negative. Xpert assay could detect 100% positive cases and RT-PCR test could detect 84.37% positive cases. Out of the 32 samples which were positive by Xpert assay, 5 (15.62%) samples had a Ctvalue greater than 40. Conclusion: The Xpert assay found to be useful as a point-ofcare test in acute scenario, where rapid and authentic diagnosis is essential, but do not have expertise and infrastructure to perform RT-PCR. [ABSTRACT FROM AUTHOR]

Abstract This study aimed to investigate the demographic, clinical, and epidemiological statistics of Japanese patients attending a designated outpatient clinic for COVID‐19 in Sapporo, Japan, and contrast the clinical and epidemiological features between those with and without mild COVID‐19. A total of 27 (8.6%) of 315 patients were diagnosed with COVID‐19. They had higher proportions of myalgia, direct contact with a confirmed COVID‐19 patient, and attendance of social gatherings in close confines. We believe that our study makes a significant contribution to the literature because it provides a clinical picture of mild COVID‐19 in the Japanese population, which has not been studied extensively. It can also assist in optimizing the local preventive measures to reduce the transmission of COVID‐19.

Introduction: Real time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) test, the gold standard test for Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) detection, is a tedious process and requires proficient workforce. Accurate and fast test results may permit more efficient use of protective and isolation resources and allow rapid therapeutic interventions. Aim: To evaluate the analytical performance characteristics of the Cepheid Xpert Xpress SARS-CoV-2 test, a rapid, automated molecular test for SARS-CoV-2 with gold standard RT-PCR test. Materials and Methods: This retrospective cohort study was conducted in Virus Research and Diagnostic Laboratory (VRDL) in Department of Microbiology at GGS Medical College, Faridkot from January 2021-June 2021. A total of 100 nasopharyngeal samples, collected from clinically suspected Coronavirus Diseae-2019 (COVID-19) cases admitted at GGSMC during 1st January-30th June 2021 were tested both by Xpert assay and RT-PCR test simultaneously, taking RT-PCR as the gold standard test. The data was analysed by MedCalc® statistical software version 19.6.4., and sensitivity, specificity, predictive values, likelihood ratios and the agreement between the two tests were calculated. Results: The mean age of the study participants was 46 years. Of these, 55 were males and 45 were females. The overall sample sensitivity and specificity of the Xpert assay were both 100% and there was perfect agreement across specimens, if authors, set a cut-off Cycle threshold-value (Ct-value) at 40 cycles for Xpert. Of 100 samples, 32 were positive for SARS-CoV-2 by either of the tests and 68 were negative. Xpert assay could detect 100% positive cases and RT-PCR test could detect 84.37% positive cases. Out of the 32 samples which were positive by Xpert assay, 5 (15.62%) samples had a Ct-value greater than 40. Conclusion: The Xpert assay found to be useful as a point-of-care test in acute scenario, where rapid and authentic diagnosis is essential, but do not have expertise and infrastructure to perform RT-PCR.

Background: COVID-19 pandemic continues to be a public health threat. Rapid antigen tests (RATs) for the detection of SARS-CoV-2 infection will help in formulation of clinical and public health strategies for the control of transmission. In India, 49% of COVID-19 tests done are RATs.¹ The sensitivity of RAT is 50.6–84%. RAT has specificity ranging from 99.3% to 100%.² The present study aims at evaluating the RAT screening test done on people attending fever clinic of Shimoga Institute of Medical Sciences, Shivamogga. Aims and Objectives: The objectives of the study were to evaluate the COVID-19 RAT screening test with reverse transcriptase-polymerase chain reaction (RT-PCR) as the gold standard test done at fever clinic of SIMS, Shimoga. Materials and Methods: An observational, cross-sectional analytical study was conducted for a period of 1 month, May 2021. The study participants included all the people attending fever clinic of SIMS, Shimoga. Assuming the sensitivity of RAT to be 85%, power of 80%, and precision of 5%, the calculated sample size was 204. Considering non-response rate of 10%, the final sample size was 224. Secondary data regarding contact number of the people attending fever clinic were collected from the COVID-19 test register. Oral consent was taken after explaining about study and assuring confidentiality. Telephonic interview was done to collect relevant information. Analysis was done using Epi Info software version 7.2.4.0. Descriptive statistics such as percentages and analytical statistics such as Student’s t-test and Chi-square test were used. Results: Overall positivity rate was 43.7%. About 71% of people had contact history. Sensitivity and specificity of RAT test were found to be 64.2% and 97.2%, respectively, and were comparable with the previous studies. Significant difference was found (P<0.05) between RAT and RT-PCR results. Conclusion: Significant difference was found between RAT and RT-PCR results which indicate that RAT is not diagnostic for people who test positive in RT-PCR. Sensitivity of RAT is relatively less in our study (but crucial in detecting disease early) and hence we strongly recommend that RT-PCR for those who test negative for RAT test. [ABSTRACT FROM AUTHOR]

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) highlights the importance of rapid diagnostic testing to identify individuals with SARS‐CoV‐2 infections and to limit the spread of the virus. Many molecular assays have become commercially available to cope with this surging demand for timely diagnosis of COVID‐19 cases, but identifying individuals requires accurate diagnostic tools. We compared the performance of three molecular SARS‐CoV‐2 assays: Aptima™ SARS‐CoV‐2 assay running on the Panther system (Hologic), an in‐house assay (Laboratory Developed Test, LDT) running on the Fusion module of the Panther Fusion system (LDT‐Fusion; Hologic), and the R‐GENE® SARS‐CoV‐2 assay (bioMérieux). In addition, we also evaluated the turnaround time. This parameter is crucial to managing the SARS‐CoV‐2 diagnosis and represents a key point in the quality management at the laboratory. Aptima™ and LDT‐Fusion assays exhibited an excellent positive percent agreement (PPA) (100.0%), while the R‐GENE® assay showed a slightly decreased PPA (98.2%). The Hologic assays have a higher throughput with less hands‐on time than the R‐GENE® assays (24–25 vs. 71 min). Both Hologic assays are used on a fully automated random‐access testing system with on‐demand testing capabilities that avoid run series, unlike the R‐GENE® assay. Automated random‐access testing systems should be preferred during periods of high SARS‐CoV‐2 prevalence. [ABSTRACT FROM AUTHOR]

The clinical presentation of COVID-19 is varied: from asymptomatic to severe neurological syndrome like stroke can happen. Guillain-Barré syndrome (GBS) as a manifestation of COVID-19 is not very common. GBS is an acute immune-mediated polyradiculoneuropathy that usually occurs following previous exposure to infection. Here, we are reporting a case of GBS related to COVID-19 infection. The reported case presented with quadriparesis and was diagnosed with GBS after evaluation. At the same time, his RT-PCR for COVID-19 was also positive. Interestingly, this patient suffered from COVID-19 2 months before this presentation. He was successfully treated with intravenous immunoglobulin. The clinician should be aware of severe neurological complications such as GBS as a potentially life-threatening complication related to COVID-19.

Background and Aim: Globally, pestiviruses are among the most economically important viral pathogens of livestock. The genus Pestivirus comprises four species, including bovine viral diarrhea virus type 1 and 2 (BVDV-1 and BVDV-2), which infect cattle, border disease virus and classical swine fever virus which infect small ruminants and pigs, respectively. Accumulating evidence suggests that pestiviruses are no longer species-specific, creating new challenges for disease control. In Indonesia, investigations related to pestiviruses remain focused on cattle as the primary host and no research has been conducted on small ruminants (sheep and goats). Therefore, the present study aimed to study the possible occurrence of pestivirus (BVDV or BVD) infections in small ruminants in Indonesia, particularly in Central Java. Materials and Methods: We used 46 blood samples consisting of 26 sheep's blood and 20 goat's blood. Samples were selected from 247 small ruminant blood collected between July and October 2020 in Central Java, Indonesia, which met the following criteria: Female, local species, approximately 1-2 years old, never been pregnant, raised in the backyard, and had no close contact with cattle in either shelter or grazing area. We tested plasma samples from sheep and goats using competitive antibody enzyme-linked immunosorbent assay to detect specific antibodies against pestivirus followed by reverse transcription-polymerase chain reaction (RT-PCR) analysis for all positive samples to differentiate the species of pestivirus. Results: Two of the 20 samples collected from goats were positive for pestivirus at the serological and molecular levels, whereas 2 of 26 samples collected from sheep were doubtful but tested negative by RT-PCR. The genotyping test results obtained using nested PCR revealed that the positive samples collected from goats had a BVDV-1 genotype. Conclusion: The results of the present study demonstrated that BVDV-1 can infect species other than bovines, in Central Java, Indonesia. Further studies involving a larger number of samples are required to: (1) Determine the actual seroprevalence of pestiviruses in small ruminants and (2) Determine the potency of small ruminants as reservoirs for pestiviruses, both of which are important for the identification of the appropriate control program for pestiviruses in Indonesia.

Breathlessness is a common presenting symptom in the emergency room. Focused history, clinical examination, and proper stepwise evaluations are essential for early diagnosis and treatment. The most common presenting feature of nCovid19 pneumonia is rapidly progressive breathlessness. During this pandemic, we need to differentiate nCovid19 pneumonia from other causes for breathlessness. Here, we discuss the differential diagnosis for rapidly progressive breathlessness, how to evaluate a patient with suspected nCovid19 pneumonia and the role of chest computed tomography scan in nCovid19.

Liver transplant recipients may face an unusually high risk for coronavirus disease 2019 (COVID-19). Observations of heightened risk, rapid progression of severe complications, greater infectivity, and potential for atypical disease presentations in transplant recipients underscore the critical importance of establishing an early diagnosis. Existing diagnostic approaches are marred by unreasonably high false-negative rates. Given the concerns for false-negative results, we performed a narrative review in effort to compile evidence for and against an optimal diagnostic algorithm for detecting COVID-19 in liver transplant recipients. In this algorithm, patients are triaged according to risk of severe acute respiratory syndrome coronavirus 2 infection. Initial testing is performed with reverse transcriptase-polymerase chain reaction, followed by chest computed tomography after 4 days. Repeat tests are performed as per the risk category, patient status, and urgency of transplant. Liver transplant centers should validate the algorithm presented herein, which is based on existing evidence and designed to maximize patient and provider safety, while assuring accuracy in diagnosis.

Background: The corona virus disease-2019 (COVID-19) or corona pandemic is an outgoing pandemic caused by coronavirus 2019 (COVID-19) caused by the transmission of severe acute respiratory syndrome coronavirus-2. It was first identified in Wuhan, China, and the first COVID-19 case in Bangladesh was on March 7, 2020. A retrospective research was conducted on Brahmanbaria Medical College with COVID-19-suspected patients to understand the current situation of COVID-19 in Brahmanbaria. Methods: A total 752 oropharyngeal and nasopharyngeal swab samples were collected from COVID-19 suspected patients and reverse transcriptase–polymerase chain reaction test was run to identify the positive cases. Results: We found 28.5% (n = 214) of positive cases among which 22.9% (n = 49) were diabetes mellitus patients and 20.5% (n = 46) were hypertension patients too. Conclusion: A total of 214 (28.5%) COVID-19 positive confirmed cases among 752 COVID-19-suspected peoples were found. The most common age group of COVID-19 patients was between 31 and 40 years, which is a matter of great concern as these patients develop different postcorona syndromes such as weakness, breathlessness, and muscles pains and cannot go back to their normal daily life as before.

Background: Although coronavirus disease-2019 (COVID-19) is predominantly a respiratory disease, cardiac involvement occurs commonly, especially in those with more severe illness. Echocardiography is the preferred imaging modality for diagnosing cardiac involvement in COVID-19. However, there are currently no data to describe echocardiographic abnormalities in Indian patients with COVID-19. Methodology: A cross-sectional observational analysis was performed among adult patients admitted to a tertiary care center between May 2020 and August 2020. Patients were included if they underwent transthoracic echocardiography during the hospitalization after a positive reverse transcriptase–polymerase chain reaction diagnosis for COVID-19 pneumonia. Demographic and clinical data were obtained and analyzed along with echocardiographic data. Results: During the study period, consecutive 245 patients were evaluated with echocardiography, of whom 11 were excluded due to nondiagnostic images. The remaining 234 (mean age 57 ± 16 years, 71.7% of men) were included in this analysis. All patients were admitted to intensive care unit or high-dependency unit. Right ventricular (RV) dilatation and/or dysfunction (37%) was the most common finding, followed by left ventricular (LV) systolic and diastolic dysfunction (27.7% and 23.1%, respectively). Pericardial effusion was present in 12% of cases. A total of 49 (20.9%) patients had preexisting LV systolic dysfunction (LVSD). After excluding them, the LVSD and LV diastolic dysfunction were observed in 8.6% and 2.7% of patients, respectively. Conclusions: This study demonstrates that RV dilatation/dysfunction is the most common echocardiographic abnormality in hospitalized patients with severe COVID-19. Further, larger, multicentric studies with systematic data collection and comparison with non-COVID patients are needed to determine the true incidence of echocardiographic abnormalities in COVID-19.

This study aimed to investigate the demographic, clinical, and epidemiological statistics of Japanese patients attending a designated outpatient clinic for COVID‐19 in Sapporo, Japan, and contrast the clinical and epidemiological features between those with and without mild COVID‐19. A total of 27 (8.6%) of 315 patients were diagnosed with COVID‐19. They had higher proportions of myalgia, direct contact with a confirmed COVID‐19 patient, and attendance of social gatherings in close confines. We believe that our study makes a significant contribution to the literature because it provides a clinical picture of mild COVID‐19 in the Japanese population, which has not been studied extensively. It can also assist in optimizing the local preventive measures to reduce the transmission of COVID‐19. [ABSTRACT FROM AUTHOR]

The diagnostic gold standard for Coronavirus-2019 disease (CoViD-19) is reverse transcriptase-polymerase chain reaction (RT-PCR). However, its sensitivity might be suboptimal. The current study aims to investigate predictive factors for false-negative nasopharyngeal RT-PCR in CoViD-19 patients. Additionally, the specificity and sensitivity of RT-PCR on the nasopharyngeal swab, serology and chest computerized-tomography (CCT) as a screening tool for the diagnosis of CoViD-19 were investigated. Medical records of patients admitted at the university hospital UZ Brussel during the CoViD-19 epidemic were reviewed. A group of CoViD-19 patients with false-negative RT-PCR was identified through scrupulous examination of medical records. Serological testing was performed through chemiluminescent microparticle assay. Eighteen CoViD-19 patients with 'false negative' RT-PCR were identified and compared to 51 'true positives'. Logistic regression for prediction of 'false negative' RT-PCR found significantly higher serology results at hospitalization and more intensive care unit admission in the group with false-negative testing. In a cohort of 228 patients, the sensitivity of RT-PCR for the diagnosis of CoViD-19 was 85%. The sensitivity of serology was 86% and its specificity 92%. Chest computerized-tomography (CCT) showed a sensitivity of 93%, its specificity was 62%. By combining RT-PCR and serology results any 'false negative' could be excluded. In this cohort, the sensitivity and specificity of RT-PCR and serology for the diagnosis of CoViD-19 were high and comparable. CCT had the highest sensitivity and confirmed its efficacy as a screening tool. CoViD-19 patients, who have a more severe presentation, might have negative RT-PCR and positive serology results. [ABSTRACT FROM AUTHOR]

Introduction: The persistence positivity detected for severe acute respiratory syndrome coronavirus 2 ribonucleic acid by real-time reverse transcriptase-polymerase chain reaction test in asymptomatic coronavirus disease 2019 positive patients has attracted a lot of attention. There is limited data on the duration of viral shedding. We aimed to determine the proportion of coronavirus disease patients with persistent positivity of real-time reverse transcriptase-polymerase chain reaction test in a teaching hospital of Nepal. Methods: A descriptive cross-sectional study was conducted using medical records from May to September 2020 in a teaching hospital of Nepal. The study was approved by the Institutional Review Committee of Karnali Academy of Health Sciences (Reference no 077/078/03). Convenient sampling method was used. Data was analysed by Statistical Package for the Social Sciences. Point estimate at 90% Confidence Interval was calculated along with frequency and proportion for binary data. Results: Of the total 95 cases, 9 (9.5%) cases (4.6-14.4 at 90% Confidence Interval), were repeat positive after achieving the first negative. The mean day required of achieving the last negative for the repeat positive group was 62.11±3.95, range (60-70 days). The mean time duration for the virus shedding was found to be 20.43±12.19 days (range 7-60 days) after the first positive test result. Conclusions: This study concludes that there might be a persistent positivity of the polymerase chain reaction test among patients with COVID-19. The majority of the patients were test positive for 8-14 days, and some were positive till 60-70 days.

Introduction: Chest Computed Tomography (CT) scan for Coronavirus Disease-2019 pneumonia is used widely and viral load is predicted by the Cycle threshold (Ct) values of Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). Studies correlating viral load in Severe Acute Respiratory SyndromeCoronavirus-2 (SARS-CoV-2) RT-PCR positive patients and severity of chest CT scan are limited. Aim: To find an association between viral load and chest CT findings. Materials and Methods: This was a cross-sectional study conducted on patients admitted to Believers Church Medical College Hospital, Thiruvalla, Kerala between September 2020 to March 2021. TrueNat RT-PCR test was performed on nasopharyngeal swabs, targeting the Orf1 gene of SARS-CoV-2 and results were quantified as Cycle threshold (Ct) values. Chest CT-Total Severity Score (TSS) ranged from 0-40 and was calculated by summing up the acute inflammatory lesions in each of the five lobes of both the lungs. Correlation was assessed using Spearman's Correlation Coefficient. Independent sample t-test and one-way Analysis of Variance (ANOVA) were used for comparison of means. Results: Of the 102 patients in the study [mean age of the patients 64.13±13.17 years and majority were males (70.6%)], 11 had lung changes unrelated to COVID-19. There was an inverse relationship between viral load Ct value of SARS-CoV-2 in nasopharyngeal specimens and TSS of chest CT scan. The mean viral load was highest in patients with mild (21.48±8.31), moderate (21.22±6.30) and severe (24.19±4.67) CT involvement. There was a significant difference between mean duration to symptom onset and chest CT scan among those with a high viral load (4.97±2.65) compared to those with a low viral load (6.81±4.5), (p-value=0.01). Among those who died due to COVID-19, (12/13) 92.3% were above 60 years of age. Presence of co-morbidities/dyspnoea/fever at presentation did not have any significant association with TSS severity. Conclusion: Viral load is not a critical factor that influences pulmonary manifestations in COVID-19, nor in-hospital mortality. CT scan may be more useful to detect lung involvement when done nearing or after the first week of symptom onset, irrespective of the viral load. Viral load can be important in predicting transmissibility and to minimise potential spread, whereas chest CT can help identify cases requiring extensive medical care.

Introduction: The emergence of severe acute respiratory syndrome coronavirus-2 pandemic is critically challenging the whole world. The real-time reverse transcriptase-polymerase chain reaction is the most widely used confirmatory test for COVID-19 detection. This study aimed to find out the prevalence of COVID-19 infection detected by gold standard reverse transcriptase-polymerase chain reaction test in a tertiary care center of Nepal. Methods: A descriptive cross-sectional study was conducted in Karnali Academy of Health Sciences from May to August 2020 after taking ethical approval from the Institutional Review Committee of Karnali Academy of Health Sciences, Jumla. Convenient sampling was used. A total of 361 participants enrolled in this study who have done real-time reverse transcriptase-polymerase chain reaction for screening of COVID-19 infection. Also, a designated questionnaire was obtained from persons with a travel history and close contact. Statistical Package for the Social Sciences software was used for the statistical analysis. Point estimate at 95% Confidence Interval was calculated along with frequency and proportion for binary data. Results: The prevalence of COVID-19 was 167 (46.3%) (95% Confidence Interval= 41.16-51.44) by real-time reverse transcriptase-polymerase chain reaction test. Out of 361 samples, 339 (93.9%) were male and 22 (6%) were female. The highest frequency of the participants belongs to the age groups of 20-40 years. Conclusions: The findings showed a high prevalence of COVID-19 detected by reverse transcriptase-polymerase chain reaction test. Further studies are necessary to improve the precision of prevalence estimations.

The clinical presentation of COVID-19 is varied: from asymptomatic to severe neurological syndrome like stroke can happen. Guillain-Barré syndrome (GBS) as a manifestation of COVID-19 is not very common. GBS is an acute immune-mediated polyradiculoneuropathy that usually occurs following previous exposure to infection. Here, we are reporting a case of GBS related to COVID-19 infection. The reported case presented with quadriparesis and was diagnosed with GBS after evaluation. At the same time, his RT-PCR for COVID-19 was also positive. Interestingly, this patient suffered from COVID-19 2 months before this presentation. He was successfully treated with intravenous immunoglobulin. The clinician should be aware of severe neurological complications such as GBS as a potentially life-threatening complication related to COVID-19. [ABSTRACT FROM AUTHOR]

Liver transplant recipients may face an unusually high risk for coronavirus disease 2019 (COVID-19). Observations of heightened risk, rapid progression of severe complications, greater infectivity, and potential for atypical disease presentations in transplant recipients underscore the critical importance of establishing an early diagnosis. Existing diagnostic approaches are marred by unreasonably high false-negative rates. Given the concerns for false-negative results, we performed a narrative review in effort to compile evidence for and against an optimal diagnostic algorithm for detecting COVID-19 in liver transplant recipients. In this algorithm, patients are triaged according to risk of severe acute respiratory syndrome coronavirus 2 infection. Initial testing is performed with reverse transcriptase-polymerase chain reaction, followed by chest computed tomography after 4 days. Repeat tests are performed as per the risk category, patient status, and urgency of transplant. Liver transplant centers should validate the algorithm presented herein, which is based on existing evidence and designed to maximize patient and provider safety, while assuring accuracy in diagnosis. [ABSTRACT FROM AUTHOR]

Although the sensitivity of reverse transcriptase-polymerase chain reaction (RT-PCR) is low in the diagnosis of coronavirus disease 2019 (COVID-19), it is the gold standard. Clinical improvement is prioritized in the follow-up of patients with COVID-19 who are followed as possible or definitive cases. Although the priority in the discharge decision is the resolution of complaints, it is also important to see radiological improvement and RT-PCR negativity. A total of 2 of our patients who were hospitalized and treated in our clinic with a diagnosis of COVID-19 were discharged after their complaints were resolved and their treatment was completed. The patients had 2 negative RT-PCR results at discharge. Both of them presented to the hospital with symptoms such as fever, cough, and shortness of breath after the discharge, and both showed positive RT-PCR results. Considering recurrent COVID-19 infection, we aimed to present treatment and the 2 cases we followed. [ABSTRACT FROM AUTHOR]

Background and Aim: Avian infectious bronchitis virus (IBV) frequently infects broilers and is responsible for severe economic losses to the poultry industry worldwide. It has also been associated with kidney damage in the broiler flocks. The aim of the present study is to determine the presence of IBV and its possible involvement in kidney damage of broiler chicks. Materials and Methods: 14 clinically diseased broiler flocks from Western and Central Algeria were sampled and analyzed by hemagglutination inhibition (HI) test and reverse transcriptase-polymerase chain reaction (RT-PCR) followed by phylogenic analysis. Results: The QX (100%) and 4/91 (60%) IBV serotypes were the most prevalent in the kidney damaged broilers regardless of vaccination status. The molecular detection of avian IBV by RT-PCR identified six samples as positive, of which only two isolates were typable by sequencing. We identified a novel IBDZ13a genotype which showed 93% sequence homology to the partial-S1 gene sequence of the IB 4/91 commercial vaccine strain. Sequencing analysis characterized this virus as a novel and divergent IB 4/91 field virus with eight amino acid substitutions that might have resulted in altered immunogenicity. Conclusion: The isolation of a new IBV strain (IBDZ13a) from vaccinated broiler flocks may explain the failure of the vaccination programs against IBV field strains. Combination of the HI test and RT-PCR indicated that the nephropathogenic IB outbreaks in broilers are related to this novel strain.

Background/Aim: To investigate the clinical and prognostic significance of MUC1 expression in patients with esophageal squamous cell carcinoma (ESCC) after radical resection. Materials and Methods: A total of 108 ESCC specimens were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) to detect MUC1 at the mRNA level and were evaluated by immunohistochemistry (IHC) to detect MUC1 at the protein level. Results: MUC1 mRNA was found in 74 cases by RT-PCR and MUC1 protein expression was found by IHC in 70 cases. Both MUC1 mRNA and protein expression correlated with pT (

Introduction: Chromosomal aberrations play a significant role in the pathogenesis of acute lymphoblastic leukemia (ALL) with prognostic and therapeutic implications. Despite the availability of molecular tools, low-resource settings struggle to diagnose the disease due to limited diagnostic capacity. The objective of this study was to detect common chromosomal aberrations in patients with ALL attending the University Teaching Hospital (UTH) in Lusaka, Zambia. Materials and Methods: In this prospective study, 19 blood samples from patients with ALL were screened for the presence of BCR-ABL, E2A-PBX1, MLL-AF4, and ETV6-RUNX1 fusion oncogenes using reverse transcriptase-polymerase chain reaction assay. Blood counts and clinical characteristics of patients were also assessed. Results: The age of patients ranged from 1½ to 72 years and comprised 57.9% of males and 42.1% of females. The majority of these patients were children (68%), and adults only comprised 32%. Only BCR-ABL and E2A-PBX1 oncogenes were detected in 3/19 of cases. The BCR-ABL gene was detected in a 4-year-old female child and a 15-year-old child. Both cases were associated with hepatomegaly and anemia coupled with low hemoglobin, white blood cell, and platelet counts. E2A-PBX1 was detected in a 12-year-old child with lymphadenopathy and splenomegaly, coupled with low hemoglobin, white blood cell, and platelet counts. All the three patients who harbored these fusion oncogenes died. Conclusion: This is the first study from Zambia to investigate the presence of fusion oncogenes in leukemia patients, which were found only among the older children population. Based on these findings, we recommend that molecular diagnosis be made a priority for the younger leukemia patient population at UTH. [ABSTRACT FROM AUTHOR]

Background: Coronavirus disease-19 (COVID-19) is a zoonotic viral disease caused by nonsegmented, enveloped, positive-sense, single-strand ribonucleic acid coronavirus. Recent outbreak started in Wuhan, China, where a new type of coronavirus was isolated from respiratory samples such as bronchoalveolar lavage and sputum of patients developing respiratory symptoms. The World Health Organization declared COVID-19 a pandemic on January 20, 2020. On April 6, 1,288,080 were infected with this virus with 70,567 deaths. Computerized tomography (CT) is the investigation of choice for diagnosing, managing, and accessing temporal changes in COVID-19. Objective: The objective of this study is to describe the chest CT findings in documented nCovid-19-positive patients. Methodology: This was a retrospective observational study done in Government Medical College, Chest Disease Hospital from February 20 to April 25, 2020. Forty-eight patients with COVID-19 reverse transcriptase-polymerase chain reaction-positive test were scanned on 64 slice Somatom CT scanner and findings analyzed. All patients with previously underlying chest disease were excluded. Results: The various chest findings in the nCovid-19-positive patients include ground glassing, (81.25%), consolidation (56.25%), nodules (43.75%), halo sign (31.25%), crazy paving pattern (50%), air bronchogram (12.5%), air bubble sign (6.25%), vascular enlargement (25%), reversed halo sign or atoll sign (18.75%), bronchial wall thickening (6.25%), and mosaic attenuation (6.25%). None of the patients had pleural effusion. Conclusion: Characteristic CT findings of COVID-19 can help radiologists in the early diagnosis of symptomatic patients in whom testing is awaited. Bilateral peripheral ground-glass opacities with consolidation in dependent parts of the lung along with the absence of pleural effusion were the most common abnormality. [ABSTRACT FROM AUTHOR]

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Congenital Zika Syndrome (CZS) is a unique pattern of congenital abnormalities found in fetuses and neonates infected with the Zika virus (ZIKV). Here, we clinically identify and characterize infants with CZS between 2015 and 2018 in Mato Grosso do Sul, Brazil—a border area with Paraguay and Bolivia. This cross-sectional study, based on primary and secondary data, tracks the cases registered in the Brazilian Public Health Reporting System through the following stages: (1) preliminary data analysis, (2) identification of the congenital syndrome cases, (3) etiologic classification of the cases, (4) active search, and (5) clinical assessment. Of the 72 investigated cases, 16 were probable cases of CZS. Of these, it was only possible to clinically assess 11 infants. Considering the 16 probable cases of CZS, nine were classified as confirmed cases, and five as potential cases of the syndrome. Regarding clinical features, brain palsy was identified in all analyzed infants. Moreover, microcephaly and pseudobulbar syndrome were found in eight infants, and hydrocephalus was found in three individuals. In addition to these conditions, seven children were malnourished. Our study may provide significant insights for other researches that aim to elucidate CZS and its clinical and populational consequences. [ABSTRACT FROM AUTHOR]

The first genotyping data on measles virus (MeV) strains in Cameroon dates from 1994, while other studies were realized in 2001 and 2011 with the establishment of MeV virological surveillance. However, the genetic data of MeV strains circulating in Cameroon remains fragmented and concentrated in certain regions, hence the need for an update. The objective of this study was to have recent data on MeV genotypes circulating in Cameroon. Ninety throat swabs collected during recent measles outbreaks were analyzed by MeV genotyping RT-PCR using the nucleoprotein gene N. The resulting sequences were analyzed on the basis of 450 nucleotides with MEGA 7 software. Overall genome analysis was performed on 40/90 sequences. The strains were from all ten regions and all belonged to cluster 1 of genotype B3. The genotype B3 has been circulating in Cameroon for long periods of time; efforts must be made in immunization for its elimination. [ABSTRACT FROM AUTHOR]

The equine hoof inner epithelium is folded into primary and secondary epidermal lamellae which increase the dermo-epidermal junction surface area of the hoof and can be affected by laminitis, a common disease of equids. Two keratin proteins (K), K42 and K124, are the most abundant keratins in the hoof lamellar tissue of Equus caballus. We hypothesize that these keratins are lamellar tissue-specific and could serve as differentiation- and disease-specific markers. Our objective was to characterize the expression of K42 and K124 in equine stratified epithelia and to generate monoclonal antibodies against K42 and K124. By RT-PCR analysis, keratin gene (KRT) KRT42 and KRT124 expression was present in lamellar tissue, but not cornea, haired skin, or hoof coronet. In situ hybridization studies showed that KRT124 localized to the suprabasal and, to a lesser extent, basal cells of the lamellae, was absent from haired skin and hoof coronet, and abruptly transitions from KRT124-negative coronet to KRT124-positive proximal lamellae. A monoclonal antibody generated against full-length recombinant equine K42 detected a lamellar keratin of the appropriate size, but also cross-reacted with other epidermal keratins. Three monoclonal antibodies generated against N- and C-terminal K124 peptides detected a band of the appropriate size in lamellar tissue and did not cross-react with proteins from haired skin, corneal limbus, hoof coronet, tongue, glabrous skin, oral mucosa, or chestnut on immunoblots. K124 localized to lamellar cells by indirect immunofluorescence. This is the first study to demonstrate the localization and expression of a hoof lamellar-specific keratin, K124, and to validate anti-K124 monoclonal antibodies. [ABSTRACT FROM AUTHOR]

Sexually transmitted infections (STIs) remain a worldwide problem and a severe threat to public health. The purpose of this study was to compare Aptima® Assays (Hologic®) and the Allplex™ STI Essential Assay (Seegene®) for the simultaneous detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis and Mycoplasma genitalium in clinical practice. The Aptima® assays (Hologic®) are based on a transcription-mediated amplification (TMA) method. The Allplex™ STI Essential assay (Seegene®) is based on a multiplex Real-Time PCR (RT-PCR) method. A total of 622 clinical samples from different anatomical sites were tested using both methods. A total of 88 (14.1%) and 66 (10.6%) positive samples were found for any of the TMA assays used and for the RT-PCR assay, respectively. Aptima® assays showed a slightly higher rate of positive results for all pathogens except for T. vaginalis, the results of which were similar to those obtained with Allplex™. The most commonly detected pathogen was C. trachomatis (37 samples; 5.9% using TMA assays) and the anatomical site with the highest prevalence of microorganisms was a non-urogenital site, the pharynx (27 positive samples; 4.3%). Using the Aptima® assays as reference method, the comparison showed that the average specificity of multiplex RT-PCR was 100.0% for the four pathogens. However an average sensitivity of 74.5% was observed, showing 95.2% (CI95%; 93.6–96.9) of overall concordance (κ = 0.80). In conclusion, the Aptima® assays show a higher sensitivity on a wide range of sample types compared to the Allplex™ assay. [ABSTRACT FROM AUTHOR]

Boid Inclusion Body Disease (BIBD) is a potentially fatal disease reported in captive boid snakes worldwide that is caused by reptarenavirus infection. Although the detection of intracytoplasmic inclusion bodies (IB) in blood cells serves as the gold standard for the ante mortem diagnosis of BIBD, the mechanisms underlying IB formation and the pathogenesis of BIBD are unknown. Knowledge on the reptile immune system is sparse compared to the mammalian counterpart, and in particular the response towards reptarenavirus infection is practically unknown. Herein, we investigated a breeding collection of 70 Boa constrictor snakes for BIBD, reptarenavirus viraemia, anti-reptarenavirus IgM and IgY antibodies, and population parameters. Using NGS and RT-PCR on pooled blood samples of snakes with and without BIBD, we could identify three different reptarenavirus S segments in the collection. The examination of individual samples by RT-PCR indicated that the presence of University of Giessen virus (UGV)-like S segment strongly correlates with IB formation. We could also demonstrate a negative correlation between BIBD and the presence of anti-UGV NP IgY antibodies. Further evidence of an association between antibody response and BIBD is the finding that the level of anti-reptarenavirus antibodies measured by ELISA was lower in snakes with BIBD. Furthermore, female snakes had a significantly lower body weight when they had BIBD. Taken together our findings suggest that the detection of the UGV-/S6-like S segment and the presence of anti-reptarenavirus IgY antibodies might serve as a prognostic tool for predicting the development of BIBD. [ABSTRACT FROM AUTHOR]

Zika virus (ZIKV) is a newly-identified infectious cause of congenital disease. Transplacental transfer of maternal IgG to the fetus plays an important role in preventing many neonatal infections. However, antibody transfer may also have negative consequences, such as mediating enhancement of flavivirus infections in early life, or trafficking of virus immune complexes to the fetal compartment. ZIKV infection produces placental pathology which could lead to impaired IgG transfer efficiency as occurs in other maternal infections, such as HIV-1 and malaria. In this study, we asked whether ZIKV infection during pregnancy impairs transplacental transfer of IgG. We enrolled pregnant women with fever or rash in a prospective cohort in Vitoria, Brazil during the recent ZIKV epidemic. ZIKV and dengue virus (DENV)-specific IgG, ZIKV and DENV neutralizing antibodies, and routine vaccine antigen-specific IgG were measured in maternal samples collected around delivery and 20 paired cord blood samples. We concluded that 8 of these mothers were infected with ZIKV during pregnancy and 12 were ZIKV-uninfected. The magnitude of flavivirus-specific IgG, neutralizing antibody, and vaccine-elicited IgG were highly correlated between maternal plasma and infant cord blood in both ZIKV-infected and -uninfected mother-infant pairs. Moreover, there was no difference in the magnitude of plasma flavivirus-specific IgG levels between mothers and infants regardless of ZIKV infection status. Our data suggests that maternal ZIKV infection during pregnancy does not impair the efficiency of placental transfer of flavivirus-specific, functional, and vaccine-elicited IgG. These findings have implications for the neonatal outomes of maternal ZIKV infection and optimal administration of antibody-based ZIKV vaccines and therapeutics. [ABSTRACT FROM AUTHOR]

It is generally accepted that ORF1629 is essential for baculovirus replication, which has enabled isolation of recombinant viruses in a baculovirus expression system using linearized viral DNA. ORF1629-defective viruses cannot replicate in insect cells; only recombinant virus with complete ORF1629 restoration by recombination can propagate, allowing for pure isolation and the development of bacmids for easy selection of recombinant viruses. We inadvertently found proliferation in insect cells of a bacmid lacking a complete ORF1629. PCR indicated no other viruses but a lack of complete ORF1629 in the proliferated bacmid, suggesting that the baculovirus propagated without a complete ORF1629. Lack of ORF1629 decreased the virus growth rate and yield; it also increased the occlusion body (OB) size but decreased its yield. These results were confirmed for Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV). Thus, entire ORF1629 is not essential for viral replication, though it does affect the virus growth rate, yield, and size and OB production. [ABSTRACT FROM AUTHOR]

ND7/23 cells are gaining traction as a host model to express peripheral sodium channels such as NaV1.8 and NaV1.9 that have been difficult to express in widely utilized heterologous cells, like CHO and HEK293. Use of ND7/23 as a model cell to characterize the properties of sodium channels requires clear understanding of the endogenous ion channels. To define the nature of the background sodium currents in ND7/23 cells, we aimed to comprehensively profile the voltage-gated sodium channel subunits by endpoint and quantitative reverse transcription-PCR and by whole-cell patch clamp electrophysiology. We found that untransfected ND7/23 cells express endogenous peak sodium currents that average –2.12nA (n = 15) and with kinetics typical of fast sodium currents having activation and inactivation completed within few milliseconds. Furthermore, sodium currents were reduced to virtually nil upon exposure to 100nM tetrodotoxin, indicating that ND7/23 cells have essentially null background for tetrodotoxin-resistant (TTX-R) currents. qRT-PCR profiling indicated a major expression of TTX-sensitive (TTX-S) NaV1.6 and NaV1.7 at similar levels and very low expression of TTX-R NaV1.9 transcripts. There was no expression of TTX-R NaV1.8 in ND7/23 cells. There was low expression of NaV1.1, NaV1.2, NaV1.3 and no expression of cardiac or skeletal muscle sodium channels. As for the sodium channel auxiliary subunits, β1 and β3 subunits were expressed, but not the β2 and β4 subunits that covalently associate with the α-subunits. In addition, our results also showed that only the mouse forms of NaV1.6, NaV1.7 and NaV1.9 sodium channels were expressed in ND7/23 cells that was originally generated as a hybridoma of rat embryonic DRG and mouse neuroblastoma cell-line. By molecular profiling of auxiliary β- and principal α-subunits of the voltage gated sodium channel complex, our results define the background sodium channels expressed in ND7/23 cells, and confirm their utility for detailed functional studies of emerging pain channelopathies ascribed to mutations of the TTX-R sodium channels of sensory neurons. [ABSTRACT FROM AUTHOR]

The galectin 3 binding protein (LGALS3BP, also known as 90K) is a ubiquitous multifunctional secreted glycoprotein originally identified in cancer progression. It remains unclear how 90K functions in innate immunity during viral infections. In this study, we found that viral infections resulted in elevated levels of 90K. Further studies demonstrated that 90K expression suppressed virus replication by inducing IFN and pro-inflammatory cytokine production. Upon investigating the mechanisms behind this event, we found that 90K functions as a scaffold/adaptor protein to interact with TRAF6, TRAF3, TAK1 and TBK1. Furthermore, 90K enhanced TRAF6 and TRAF3 ubiquitination and served as a specific ubiquitination substrate of TRAF6, leading to transcription factor NF-κB, IRF3 and IRF7 translocation from the cytoplasm to the nucleus. Conclusions: 90K is a virus-induced protein capable of binding with the TRAF6 and TRAF3 complex, leading to IFN and pro-inflammatory production. [ABSTRACT FROM AUTHOR]

C-to-U editing is an important event in post-transcriptional RNA processing, which converts a specific cytidine (C)-to-uridine (U) in transcripts of mitochondria and plastids. Typically, the pentatricopeptide repeat (PPR) protein, which specifies the target C residue by binding to its upstream sequence, is involved in the editing of one or a few sites. Here we report a novel PPR-DYW protein EMP21 that is associated with editing of 81 sites in maize. EMP21 is localized in mitochondria and loss of the EMP21 function severely inhibits the embryogenesis and endosperm development in maize. From a scan of 35 mitochondrial transcripts produced by the Emp21 loss-of-function mutant, the C-to-U editing was found to be abolished at five sites (nad7-77, atp1-1292, atp8-437, nad3-275 and rps4-870), while reduced at 76 sites in 21 transcripts. In most cases, the failure to editing resulted in the translation of an incorrect residue. In consequence, the mutant became deficient with respect to the assembly and activity of mitochondrial complexes I and V. As six of the decreased editing sites in emp21 overlap with the affected editing sites in emp5-1, and the editing efficiency at rpl16-458 showed a substantial reduction in the emp21-1 emp5-4 double mutant compared with the emp21-1 and emp5-4 single mutants, we explored their interaction. A yeast two hybrid assay suggested that EMP21 does not interact with EMP5, but both EMP21 and EMP5 interact with ZmMORF8. Together, these results indicate that EMP21 is a novel PPR-DYW protein required for the editing of ~17% of mitochondrial target Cs, and the editing process may involve an interaction between EMP21 and ZmMORF8 (and probably other proteins). [ABSTRACT FROM AUTHOR]

Background: The cocirculation of duck hepatitis A virus subtypes 1 (DHAV-1) and 3 (DHAV-3) in ducklings has resulted in significant economic losses. Ducklings with DHAV-1 or DHAV-3 infection show similar clinical signs and gross lesions; hence, it is important to identify the viral subtypes in infected ducklings as early as possible for better clinical management. Methods and results: Based on multiple 5’ noncoding region (5’-NCR) sequences of DHAV-1 and DHAV-3 strain alignments, universal and type-specific primers were designed and synthesized. With three primers in one-tube reverse transcription-PCR (RT-PCR), reference DHAV-1 and DHAV-3 isolates ranging over 60 years and across many different countries were successfully amplified, indicating that the primer sequences were completely conserved. The sequence results and the sizes of amplicons from reference DHAV-1 and DHAV-3 isolates are completely correlated with their subtypes. Moreover, with this one-tube RT-PCR system, amplicon sizes from liver samples of reference DHAV-1- or DHAV-3-infected birds fit closely with their subtypes, which was determined by virus isolation and neutralization testing. No other duck-origin RNA viruses were detected. The sensitivity of viral RNA detection was 10 pg. With this system, 20% subtype 1, 45% subtype 3, and 9% coinfection of two subtypes were detected in 55 clinical samples. Conclusions and significance: This novel approach could be used for rapidly typing DHAV-1 or DHAV-3 infection in routine clinical surveillance or epidemiological screening. [ABSTRACT FROM AUTHOR]

Group A rotaviruses (RVA) are one of the major causes of acute gastroenteritis (AGE) in young children worldwide. Owing to lack of proper surveillance programs and health facilities, developing countries of Asia and Africa carry a disproportionately heavy share of the RVA disease burden. The aim of this hospital-based study was to investigate the circulation of RVA genotypes in Rawalpindi and Islamabad, Pakistan in 2015 and 2016, prior to the implementation of RVA vaccine. 639 faecal samples collected from children under 10 years of age hospitalized with AGE were tested for RVA antigen by ELISA. Among 171 ELISA positive samples, 143 were successfully screened for RT-PCR and sequencing. The prevalence of RVA was found to be 26.8% with the highest frequency (34.9%) found among children of age group 6–11 months. The most predominant circulating genotypes were G3P[8] (22.4%) followed by G12P[6] (20.3%), G2P[4] (12.6%), G1P[8] (11.9%), G9P[6] (11.9%), G3P[4] (9.1%), G1P[6] (4.2%), G9P[8] (4.2%), and G3P[6] (0.7%). A single mixed genotype G1G3P[8] was also detected. The findings of this study provide baseline data, that will help to assess if future vaccination campaigns using currently available RVA vaccine will reduce RVA disease burden and instigate evolutionary changes in the overall RVA biology. The high prevalence of RVA infections in Pakistan require to improve and strengthen the surveillance and monitoring system for RVA. This will provide useful information for health authorities in planning public health care strategies to mitigate the disease burden caused by RVA. [ABSTRACT FROM AUTHOR]