关键词： SARS-CoV-2   RT-PCR   Extraction-Free  

摘要： The worldwide demand for SARS-CoV-2 RT-PCR testing resulted in a shortage of diagnostic kits. RNA extraction step constitutes a major bottleneck to perform diagnostic. The aim of this study was to assess performances of different extraction-free SARS-CoV-2 RT-PCR assays compared to a reference RT-PCR assay. The panel of evaluation consisted of 94 samples: 69 positive and 25 negative for SARS-CoV-2 by reference RTPCR. Three extraction-free RT-PCR assays were assessed: (i) PrimeDirect? Probe RT-qPCR Mix (Takara), (ii) PrimeScript?RT-PCR (Takara), and (iii) SARS-CoV-2 SANSURE?BIOTECH Novel Coronavirus (Sansure). The overall sensitivity of PrimeDirect, PrimeScript and Sansure assays was 55.1 %, 69.6 % and 69.6 %, respectively. The sensitivity increased among samples with Ct<30: 91.9 % (n = 34/37), 89.2 % (n = 33/37) and 94.6 % (n = 35/37) for PrimeDirect, PrimeScript and Sansure assays, respectively. The specificity was 88 %, 100 % and 100 % for PrimeDirect, PrimeScript and Sansure assays, respectively. In the present study, we showed a good sensitivity of extraction-free PCR assays, especially for high viral loads (Ct<30), except PrimeDirect that displayed imperfect sensitivity and specificity. Despite a lower sensitivity for low viral loads, extraction-free reagents can provide a valuable option, cheaper, easier and less reagent consuming for SARS-CoV-2 diagnostic, especially in laboratory with lower experience and equipment for molecular assays.

关键词： hip   acetabulum   hip abduction   acetabular development   hip dysplasia  

摘要： Raja Ravi Varma's painting "There Comes Papa" depicts his daughter carrying her son astride her hip. The positive implication of the posture of the hips of the child while being carried in this manner on acetabular development is discussed.

关键词： Alternative mRNA splicing   CD4 versus CD8 T cell lineages   Transcriptomics   High throughput sequencing   Illumina   PacBio  

摘要： T cells can be subdivided into a number of different subsets that are defined by their distinct functions. While the specialization of different T cell subsets is partly achieved by the expression of specific genes, the overall transcriptional profiles of all T cells appear very similar. Alternative mRNA splicing is a mechanism that facilitates greater transcript/protein diversity from a limited number of genes, which may contribute to the functional specialization of distinct T cell subsets. In this study we employ a combination of short-read and long-read sequencing technologies to compare alternative mRNA splicing between the CD4 and CD8 T cell lineages. While long-read technology was effective at assembling full-length alternatively spliced transcripts, the low sequencing depth did not facilitate accurate quantitation. On the other hand, short-read technology was ineffective at assembling full-length transcripts but was highly accurate for quantifying expression. We show that integrating long-read and short-read data together achieves a more complete view of transcriptomic diversity. We found that while the overall usage of transcript isoforms was very similar between the CD4 and CD8 lineages, there were numerous alternative spliced mRNA isoforms that were preferentially used by one lineage over the other. These alternative spliced isoforms included ones with different exon usage, exon exclusion or intron inclusion, all of which are expected to significantly alter the protein sequence.

关键词： cost implication   diagnostic algorithm   haemodialysis   HCV core antigen   hepatitis C virus  

摘要： Aims In Malaysia, majority anti-HCV positive haemodialysis patients do not undergo hepatitis C confirmation due to the high cost of HCV RNA. HCV Core Antigen might be a cost-effective diagnostic test to identify HD patients who have active HCV infection eligible for Direct Acting Anti-viral therapy. Methods A cross-sectional study was conducted to assess the correlation between HCV Ag and HCV RNA and the cost implications of different diagnostic algorithms to diagnose active HCV infection using Anti-HCV, HCV Ag, and HCV RNA. Pre-dialysis blood was tested for both HCV Ag and HCV RNA. HCV Ag was tested with Abbott ARCHITECT HCV Ag test. Results Two-hundred twenty-seven haemodialysis patients were recruited from 20 centres with mean age of 57.68 +/- 12.48 years, and male constitutes 56.8% (129) of the study population. HCV Ag correlated well with HCV RNA (Spearman test coefficient 0.943, p < .001) with sensitivity of 93.9%, specificity 99.3%, and the accuracy was 97.36%. Cost analysis indicated that a sequential test involving Anti-HCV antibody as initial screening, followed by HCV Ag on Anti-HCV positive and HCV RNA on HCV Ag negative cases translated to a modest cost-saving algorithm compared to standard diagnostic algorithm. Conclusion HCV Ag correlated well with HCV RNA and can potentially be fused in an alternative diagnostic algorithm to generate cost savings methods to diagnose active HCV infection among haemodialysis patients. This alternative algorithm is especially relevant in low to middle-income countries such as Malaysia to optimize the use of the healthcare resource and gains in clinical outcomes.

关键词： pediatric sports medicine   tibial spine fracture   arthroscopy   trauma   knee  

摘要： Background: Postoperative ipsilateral anterior cruciate ligament (ACL) tears after tibial eminence fracture fixation has been previously noted in the literature. This study aims to describe the prevalence of and risk factors for postoperative ACL tears in a cohort of patients operatively treated for tibial eminence fracture. Methods: A retrospective review of children undergoing treatment of a tibial eminence fracture at 10 tertiary care children's hospitals was performed. The primary outcome of interest was subsequent ACL rupture. Incidence of ACL tear was recorded for the entire cohort. Patients who sustained a postoperative ACL tear were compared with those without ACL tear and analyzed for demographics and risk factors. A subgroup analysis was performed on patients with a minimum of 2-year follow-up data or those who had met the primary outcome (ACL tear) before 2 years. Results: A total of 385 pediatric patients were reviewed. 2.6% of the cohort experienced a subsequent ACL tear. The median follow-up time was 6.5 months (SD=6.4 mo). Subsequent ACL tears occurred at a median of 10.2 months (SD=19.5 mo) postoperatively. There was a statistically significant association with higher grade tibial spine fractures (Myers and McKeever type III and IV) and subsequent ACL tear (P=0.01). Patients with a subsequent ACL tear were older on average (13.5 vs. 12.2 y old), however, this difference was not statistically significant (P=0.08). Subgroup analysis of 46 patients who had a 2-year follow-up or sustained an ACL tear before 2 years showed a 21.7% incidence of a subsequent ACL tear. There was a statistically significant association with higher grade tibial spine fractures (Myers and McKeever type III and IV) and subsequent ACL rupture (P=0.006) in this subgroup. Postoperative ACL tears occurred in patients who were older at the time that they sustained their original tibial eminence fracture (13.4 vs. 11.3 y old, P=0.035). Conclusions: Ipsilateral ACL tears following oper

关键词： Substance use disorder   Personality   Neurobehavioral   Disinhibition   Conscientiousness   Co-twin control  

摘要： To establish a trait-dispositional variable as an indicator of liability for the development of substance use disorders (SUDs), the trait must share heritable variance with SUDs and its association should not be primarily attributable to a direct impact of SUDs on characteristics that define the trait. The current work applied a co-twin control (CTC) modeling approach to data from two monozygotic twin samples to investigate the degree to which different measures of trait-impulsiveness represent indicants of vulnerability to SUDs (liability indicators), or outcomes or concomitants of SUDs (exposure indicators). The Five Factor Model (FFM) trait of conscientiousness was assessed via self-report, and a counterpart neurobehavioral trait of disinhibition was assessed both through self-report and using self-report and brain response measures combined. FFM trait data were available for one twin sample (N = 298);data for variants of P3 brain response were available along with a scale measure of disinhibition in the other (N = 258). CTC analyses revealed only an exposure effect of SUD symptomatology on FFM conscientiousness, indicating that this self-report assessed trait does not index liability for SUDs. By contrast, the disinhibition scale measure showed pronounced liability and weaker exposure-based associations with SUDs ? and when quantified using scale scores together with P3 brain response, the exposure-based association was eliminated, such that this disinhibition measure related to SUD symptoms exclusively as a function of liability influences. These findings highlight a distinct advantage of quantifying traits in neurobehavioral terms ? namely, the capacity to effectively index dispositional liability for psychopathological outcomes.

关键词： HuD   Elavl4 KO   alternative splicing   alternative polyadenylation   neocortex  

摘要： The neuronal Hu/ELAV-like proteins HuB, HuC and HuD are a class of RNA-binding proteins that are crucial for proper development and maintenance of the nervous system. These proteins bind to AU-rich elements (AREs) in the untranslated regions (3 '-UTRs) of target mRNAs regulating mRNA stability, transport and translation. In addition to these cytoplasmic functions, Hu proteins have been implicated in alternative splicing and alternative polyadenylation in the nucleus. The purpose of this study was to identify transcriptome-wide effects of HuD deletion on both of these nuclear events using RNA sequencing data obtained from the neocortex of Elavl4(-/-) (HuD KO) mice. HuD KO affected alternative splicing of 310 genes, including 17 validated HuD targets such as Cbx3, Cspp1, Snap25 and Gria2. In addition, deletion of HuD affected polyadenylation of 53 genes, with the majority of significantly altered mRNAs shifting towards usage of proximal polyadenylation signals (PAS), resulting in shorter 3 '-UTRs. None of these genes overlapped with those showing alternative splicing events. Overall, HuD KO had a greater effect on alternative splicing than polyadenylation, with many of the affected genes implicated in several neuronal functions and neuropsychiatric disorders.

摘要： The COVID-19 pandemic has had significant ramifications for provider well-being. During these unprecedented and challenging times, one institution's Department of Surgery put in place several important initiatives for promoting the well-being of trainees as they were redeployed to provide care to COVID-19 patients. In this article, the authors describe these initiatives, which fall into 3 broad categories: redeploying faculty and trainees, ensuring provider safety, and promoting trainee wellness. The redeployment initiatives are the following: reframing the team mindset, creating a culture of grace and forgiveness, establishing a multidisciplinary wellness committee, promoting centralized leadership, providing clear communication, coordinating between departments and programs, implementing phased restructuring of the department's services, establishing scheduling flexibility and redundancy, adhering to training regulations, designating a trainee ombudsperson, assessing physical health risks for high-risk individuals, and planning for structured deimplementation. Initiatives specific to promoting provider safety are appointing a trainee safety advocate, guaranteeing personal protective equipment and relevant information about these materials, providing guidance regarding safe practices at home, and offering alternative housing options when necessary. Finally, the initiatives put in place to directly promote trainee wellness are establishing an environment of psychological safety, providing mental health resources, maintaining the educational missions, solidifying a sense of community by showing appreciation, being attentive to childcare, and using social media to promote community morale. The initiatives to carry out the department's strategy presented in this article, which were well received by both faculty and trainee members of the authors' community, may be employed in other departments and even outside the context of COVID-19. The authors hope that colleagues a

关键词： Beef   Shelf-life   Carcass   Conventional chilling   Modified chilling   Microbiology  

摘要： The objective of this study was to compare the effect of current (10 degrees C for 10 h followed by 0 degrees C with a low fan speed) versus four alternative beef carcass chilling regimes, ranging from -6 degrees C to 0 degrees C and wind speeds between 1.5 and 6 m/s on the microbiology of beef carcasses. The temperature and relative humidity (RH) in the chillers, the carcass core and surface temperature, pH, water activity (a(w)) and carcass weight (drip) loss were recorded. Bacterial concentrations (total viable counts (TVC), total Enterobacteriaceae counts (TEC), Pseudomonas spp., lactic acid bacteria (LAB) and Brochothrix thermosphacta) were also monitored. Similar pH, a w and drip loss (2%) values were obtained regardless of chilling regime. For the most part, bacterial concentrations were also similar and, where statistically significant (P < 0.05) counts occurred, the reductions were low (<= 1 log(10) cfu/cm(2)). It was concluded that the current chilling regime was as effective as the tested alternatives in terms of the bacterial quality of the carcasses.