QTQTN motif upstream of the furin-cleavage site plays a key role in SARS-CoV-2 infection and pathogenesis

The furin cleavage site (FCS), an unusual feature in the SARS-CoV-2 spike protein, has been spotlighted as a factor key to facilitating infection and pathogenesis by increasing spike processing. Similarly, the QTQTN motif directly upstream of the FCS is also an unusual feature for group 2B coronaviruses (CoVs). The QTQTN deletion has consistently been observed in in vitro cultured virus stocks and some clinical isolates. To determine whether the QTQTN motif is critical to SARS-CoV-2 replication and pathogenesis, we generated a mutant deleting the QTQTN motif (ΔQTQTN). Here, we report that the QTQTN deletion attenuates viral replication in respiratory cells in vitro and attenuates disease in vivo. The deletion results in a shortened, more rigid peptide loop that contains the FCS and is less accessible to host proteases, such as TMPRSS2. Thus, the deletion reduced the efficiency of spike processing and attenuates SARS-CoV-2 infection. Importantly, the QTQTN motif also contains residues that are glycosylated, and disruption of its glycosylation also attenuates virus replication in a TMPRSS2-dependent manner. Together, our results reveal that three aspects of the S1/S2 cleavage site—the FCS, loop length, and glycosylation—are required for efficient SARS-CoV-2 replication and pathogenesis.

SARS-CoV-2 emerged in late 2019 and has caused the largest pandemic since the 1918 influenza outbreak (1). An unusual feature of SARS-CoV-2 is the presence of a furin cleavage site (FCS) in its spike protein (2). The CoV spike is a trimer of spike proteins composed of the S1 and S2 subunits, responsible for receptor binding and membrane fusion, respectively (1). After receptor binding, the spike protein is proteolytically cleaved at the S1/S2 and S2′ sites to activate the fusion machinery. For SARS-CoV-2, the spike protein contains a novel cleavage motif recognized by the host cell furin protease (PRRAR) directly upstream of the S1/S2 cleavage site that facilitates cleavage prior to virion release from the producer cell. This FCS, not found in other group 2B CoVs, plays a key role in spike processing, infectivity, and pathogenesis as shown by our group and others (3, 4).Importantly, another novel amino acid motif, QTQTN, is found directly upstream of the FCS. This QTQTN motif, also absent in other group 2B CoVs, is often deleted and has been pervasive in cultured virus stocks of the alpha, beta, and delta variants (5–8). In addition, the QTQTN deletion has been observed in a small subset of patient samples as well (9–11). Because this deletion has been frequently identified, we set out to characterize it and determine whether it has consequences for viral replication and virulence. Using our infectious clone (12, 13), we demonstrated that the loss of this motif attenuates SARS-CoV-2 replication in respiratory cells in vitro and pathogenesis in hamsters. The QTQTN deletion results in reduced spike cleavage and diminished capacity to use serine proteases on the cell surface for entry. Importantly, mutations of glycosylation-enabling residues in the QTQTN motif results in similar replication attenuation despite intact spike processing. Together, our results highlight elements in the SARS-CoV-2 spike in addition to the FCS that contribute to increased replication and pathogenesis.     

薄层扫描法测定藤茶中二氢杨梅素的含量

目的：建立藤茶质量控制的方法,方法：采用薄层扫描法测定藤茶中二氢杨梅素的含量。结果：藤茶中二氢杨梅素的含量３８．１７％－３８．５４％,回收率为９８．５％,ＲＳＤ为１．６％,结论：该法快速、简单、稳定、测定结果准确、可靠。     

Timely follow‐up of positive cancer screening results: A systematic review and recommendations from the PROSPR Consortium

Timely follow‐up for positive cancer screening results remains suboptimal, and the evidence base to inform decisions on optimizing the timeliness of diagnostic testing is unclear. This systematic review evaluated published studies regarding time to follow‐up after a positive screening for breast, cervical, colorectal, and lung cancers. The quality of available evidence was very low or low across cancers, with potential attenuated or reversed associations from confounding by indication in most studies. Overall, evidence suggested that the risk for poorer cancer outcomes rises with longer wait times that vary within and across cancer types, which supports performing diagnostic testing as soon as feasible after the positive result, but evidence for specific time targets is limited. Within these limitations, we provide our opinion on cancer‐specific recommendations for times to follow‐up and how existing guidelines relate to the current evidence. Thresholds set should consider patient worry, potential for loss to follow‐up with prolonged wait times, and available resources. Research is needed to better guide the timeliness of diagnostic follow‐up, including considerations for patient preferences and existing barriers, while addressing methodological weaknesses. Research is also needed to identify effective interventions for reducing wait times for diagnostic testing, particularly in underserved or low‐resource settings. CA Cancer J Clin 2018;68:199–216 . © 2018 American Cancer Society .     

Mental health consequences of intimate partner abuse: a multidimensional assessment of four different forms of abuse

Battered women are exposed to multiple forms of intimate partner abuse. This article explores the independent contributions of physical violence, sexual coercion, psychological abuse, and stalking on symptoms of posttraumatic stress disorder (PTSD) and depression among a sample of 413 severely battered, help-seeking women. The authors test the unique effects of psychological abuse and stalking on mental health outcomes, after controlling for physical violence, injuries, and sexual coercion. Mean scores for the sample fall into the moderate to severe range for PTSD and within the moderate category for depression scores. Hierarchical regressions test the unique effects of stalking and psychological abuse, after controlling for physical violence, injuries, and sexual coercion. Psychological abuse and stalking contribute uniquely to the prediction of PTSD and depression symptoms, even after controlling for the effects of physical violence, injuries, and sexual coercion. Results highlight the importance of examining multiple dimensions of intimate partner abuse.     

An assessment of water quality,physical habitat,and biological integrity of an urban stream in Wichita,Kansas, prior to restoration improvements (phase I)

Urban development alters the natural hydrological conditions of many streams and rivers often resulting in the degradation of water quality, physical habitat, and biotic integrity of lotic systems. Restoration projects attempt to improve and maintain the ecological integrity of urban streams; however, few projects have quantified improvements to stream ecology following implementation of restoration measures. This paper summarizes pre-restoration data collected as part of an urban stream restoration project on Gypsum Creek in Wichita, Kansas. Water quality monitoring revealed eutrophic conditions in the stream and the presence of pesticides. Channelization has led to changes in physical habitat including bank erosion, sedimentation, loss of substrate and channel diversity, elimination of in-stream aquatic habitat, removal of riparian vegetation, and decreased base flows. Benthic macroinvertebrate communities appear degraded with more than 90% of individuals collected described as tolerant to anthropogenic stressors. Fish communities were assessed with an Index of Biotic Integrity and were rated as poor to fair, with trophic structure dominated by generalists, no sensitive species present, and one-third of the species collected considered non-native. Overall, the data collected strongly suggest that site-specific restoration measures need to be implemented in order to improve and maintain the ecological condition of Gypsum Creek. Recommendations for improvements have been made to city managers, with implementation beginning in spring 2003 (dependent upon funding availability). Received: 28 March 2002/Accepted: 24 July 2002     

An internal fixator for posterior application to short segments of the thoracic, lumbar, or lumbosacral spine. Design and testing

A new spinal implant has been designed and biomechanical testing completed, intended for application to "short-segment" spinal defects such as disc degeneration, fracture, spondylolisthesis, or tumor. Major improvements over currently available devices include: only 2-3 vertebrae are spanned, not 5-7 as with Harrington rods; true three-dimensional fixation is achieved, preventing such problems as hook or rod dislocation; three-dimensional adjustment is easily accomplished, allowing fracture or spondylolisthesis reduction to be readily performed; attachment to vertebrae is by means of transpedicular screws eliminating deliberate encroachment into the spinal canal, such as Luque wires or Harrington hooks; no special alignment between screws is needed (such as with holes or slots in a plate), allowing screw placement to fully conform to anatomic structures; and laminectomy sites and lumbosacral junction are readily instrumented. Background investigations presented here for design of this device include: CT-defined pedicle morphometry showing that screws may be larger than those currently used; effect of pitch, minor diameter, and tooth profile on screw pull-out strength; mechanical testing of a compact, three-dimensionally adjustable, strong, nonloosening articulating clamp; and establishing of the relationship between depth of penetration and strength of fixation of transpeduncular screws.     

Chaplaincy Research: Its Value,Its Quality,and Its Future

The article is divided into four major sections, the first of which presents and discusses various reasons given by major researchers in the field why chaplains should do research. The second section summarizes findings on the sophistication of research on religion and health published in (a) medical and other healthcare journals, and (b) specialty journals on religion and health, chaplaincy, and pastoral care and counseling. The third section revisits suggestions that have been made by prominent chaplain researchers to increase and improve research by chaplains. The last section offers some suggestions for expanding several lines of current research in the future, including research: (1) to elucidate the nature of spiritual care chaplains provide to different populations, including patients, families and staff; (2) to assess the prevalence and intensity of patients' spiritual needs and the degree to which they are being met; (3) to identify that subset of patients who are spiritually at risk in terms of having high needs and slow religious resources; (4) to identify the biological causal mechanisms by which religion influences health; and (5) to measure the effectiveness of chaplain interventions.     

Minding the gap: health and social care provider perceptions of parental communication and Black-Canadian youths’ sexual health

Abstract

Health and social care providers’ perceptions of Black-Canadian parent-youth sexual health communication has important implications for addressing knowledge gaps in the provision of services to young people and their parents. Providers’ perceptions are crucial as they often act as advisers in tailoring programmes or services to the perceived needs of parents and youth. To understand these perceptions, 17 semi-structured in-depth interviews were conducted with providers who worked with African, Caribbean or Black (ACB) parents and youth in Toronto, Ontario, Canada. Critical Race Theory was used to help guide the interpretation of findings. The findings revealed providers believed that many parents were unlikely to explicitly discuss sexual health or HIV prevention with young people. Additionally, providers perceived that the content of and approach to parent-youth sexual health communication differed between African and Caribbean clients. Moreover, providers believed that both parents’ and young people’s sex and gender impacted the quality, content and style of sexual health communication and had important implications for programme development. Overall, findings suggest a need for understanding the development of providers’ perceptions of this communication, ways to address these perceptions and further parent-provider collaboration to promote Black youths’ sexual health.