Journal Article Lack of Association of Cytomegalovirus with Endemic African Kaposi's Sarcoma Get access Richard F. Ambinder, Richard F. Ambinder Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Please address requests for reprints to Dr. Richard Ambinder, 3-121 Oncology, Johns Hopkins Hospital, Baltimore, Maryland 21215. Search for other works by this author on: Oxford Academic PubMed Google Scholar Cheryl Newman, Cheryl Newman Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Gary S. Hayward, Gary S. Hayward Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Robert Biggar, Robert Biggar Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Mads Melbye, Mads Melbye Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Luc Kestens, Luc Kestens Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Eric Van Marck, Eric Van Marck Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Peter Plot, Peter Plot Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Paul Gigase, Paul Gigase Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar Pamela B. Wright, Pamela B. Wright Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar ... Show more Thomas C. Quinn Thomas C. Quinn Departments of Oncology, Infectious Disease, and Pharmacology, Johns Hopkins Medical School, Baltimore, and the National Cancer Institute and the Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland; the Institute of Cancer Research, Radiumstationen, Aarhus, Denmark; and the Departments of Pathology and Microbiology, Institute of Tropical Medicine, Antwerp, Belgium Search for other works by this author on: Oxford Academic PubMed Google Scholar The Journal of Infectious Diseases, Volume 156, Issue 1, July 1987, Pages 193–197, https://doi.org/10.1093/infdis/156.1.193 Published: 01 July 1987 Article history Received: 04 August 1986 Revision received: 29 December 1986 Published: 01 July 1987
Background The National Early Warning Scores (NEWS) is used in various healthcare settings to augment clinical decision making, and there is growing interest in its application in primary care. This research aimed to determine the distribution of NEWS among patients in UK out-of-hours (OOH) general practice and explore the relationship between NEWS and referral of patients to hospital. Methods A historical cohort study using routinely collected data from the Birmingham Out-of-hours general practice Research Database. This includes patients who attended a large out-of-hours general practice provider in the West Midlands, UK, between July 2013 and July 2018. All adults who were seen face to face who had a full set of physiological observations recorded were included. NEWS was calculated post hoc, and subsequent hospital referral was the outcome of interest. Results A NEWS was calculated for 74 914 consultations. 46.9% of patients had a NEWS of 0, while 30.6% had a NEWS of 1. Patients were referred to hospital in 8.5% of all encounters. Only 6.9% (95% CI 6.3% to 7.5%) of the 6878 patients referred to hospital had a NEWS of ≥5. Of the 1509 patients with a NEWS ≥5, 68.6% (95%CI 66.2% to 70.9%) were not referred to hospital. When considering how NEWS was related to hospital referral, the area under the receiver operating characteristic (AUROC) for patients seen in their own home was 0.731 (95%CI 0.681 to 0.787). For patient seen in treatment centres, the AUROC was 0.589 (95% CI 0.582 to 0.596). Conclusions Patients seen in out-of-hours general practice have low physiological acuity. Those referred to hospital have a slightly higher NEWS overall. NEWS is poorly associated with hospital referral, although the association is stronger for patients seen in at home compared with patients seen in treatment centres. Implementing NEWS-based referral from OOH general practice is likely to increase hospital admissions.
The COVID-19 pandemic has given rise to numerous commercially available antigen rapid diagnostic tests (Ag-RDTs). To generate and to share accurate and independent data with the global community requires multisite prospective diagnostic evaluations of Ag-RDTs. This report describes the clinical evaluation of the OnSite COVID-19 rapid test (CTK Biotech, CA, USA) in Brazil and the United Kingdom. A total of 496 paired nasopharyngeal (NP) swabs were collected from symptomatic health care workers at Hospital das Clínicas in São Paulo, Brazil, and 211 NP swabs were collected from symptomatic participants at a COVID-19 drive-through testing site in Liverpool, United Kingdom. Swabs were analyzed by Ag-RDT, and results were compared to quantitative reverse transcriptase PCR (RT-qPCR). The clinical sensitivity of the OnSite COVID-19 rapid test in Brazil was 90.3% (95% confidence interval [CI], 75.1 to 96.7%) and in the United Kingdom was 75.3% (95% CI, 64.6 to 83.6%). The clinical specificity in Brazil was 99.4% (95% CI, 98.1 to 99.8%) and in the United Kingdom was 95.5% (95% CI, 90.6 to 97.9%). Concurrently, analytical evaluation of the Ag-RDT was assessed using direct culture supernatant of SARS-CoV-2 strains from wild-type (WT), Alpha, Delta, Gamma, and Omicron lineages. This study provides comparative performance of an Ag-RDT across two different settings, geographical areas, and populations. Overall, the OnSite Ag-RDT demonstrated a lower clinical sensitivity than claimed by the manufacturer. The sensitivity and specificity from the Brazil study fulfilled the performance criteria determined by the World Health Organization, but the performance obtained from the UK study failed to do. Further evaluation of Ag-RDTs should include harmonized protocols between laboratories to facilitate comparison between settings. IMPORTANCE Evaluating rapid diagnostic tests in diverse populations is essential to improving diagnostic responses as it gives an indication of the accuracy in real-world scenarios. In the case of rapid diagnostic testing within this pandemic, lateral flow tests that meet the minimum requirements for sensitivity and specificity can play a key role in increasing testing capacity, allowing timely clinical management of those infected, and protecting health care systems. This is particularly valuable in settings where access to the test gold standard is often restricted.
Abstract Introduction: Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiological agent of KS, which is the leading cause of cancer and cancer related mortality in HIV/AIDS. African Americans currently have the highest incidence of KS throughout the world. This study examines the initial pathogenic path leading to, or promoting KS, and considers if a protein(s) in that path could provide a novel prognostic and therapeutic target for KS. Experimental Procedures: Microarray analysis was used to examine KSHV and mock infected dermal microvascular endothelial cells (DMVEC). Expression of fibulin-2 protein was examined by dual labeled immunofluorescense and immunoblot analysis. Transcriptional analysis of fibulins was analyzed by quantitative real time polymerase chain reaction (qRT-PCR). KSHV infection levels in primary effusion lymphoma cell lines (PELs) were examined by Southern blot hybridization using a 32P labeled viral genome terminal repeat fragment as a probe. Fibulin-2 protein expression in PELs was determined by immunoblots. Fibulin-2 and KSHV latency associated nuclear antigen (LANA) expression in KS tumors was performed using KS tumor tissue microarrays (from the AIDS Cancer Specimen Resource Consortium) and dual labeled immunohisto-chemistry. Transcriptional analysis of fibulin-2 ligand binding partners and time course transcriptional analysis of infected and control DMVEC cells were performed by qRTPCR. Data Summary and Conclusions: Fibulins are extra-cellular matrix proteins (ECM) involved in cell adhesion, proliferation, migration, invasion and angiogenesis, and have been linked to progression of several cancers. To our knowledge, fibulins have not been studied in KS. Here we demonstrate that fibulin-2 protein expression is down-regulated 50-fold in 10 day KSHV infected DMVEC with a 26-fold reduction in fibulin-2 message. By time-course analysis there was a consistent reduction of fibulin-2 message accompanied by an increase in LANA transcription. After examining all fibulin family members, fibulins −2, −5 and −3 were down-regulated over time in KSHV infected DMVEC, fibulins 1C and 1D were up-regulated, and no change in fibulins 4, 6 and 7. In contrast, in PELs (which express different levels of KSHV lytic replication), we found no detectable fibulin-2 expression. Tissue microarrays representing patch/plaque and nodular forms of KS from 86 patients were shown to be statistically significant for downregulation of fibulin-2 in virus infected LANA positive cells by dual labeled immunohistochemical staining. We also examined transcriptional dysregulation of fibulin-2 ligand binding partners laminin a2, fibrillin, aggrecan, versican, fibronectin, nidogen, perlecan, and tropoelastin. We found a 5 and 12.5-fold transcriptional suppression of fibronectin and tropoelastin mRNA, respectively, in virus infected cells, with no significant change in transcription for other fibulin-2 ligand binding partners. This represents the first study to examine fibulin-2 expression in KSHV infected DMVEC and in KS tissue samples to determine if suppression of this ECM protein or its ECM ligand binding partners play a role in KS tumor progression. Understanding the interactions between KSHV, fibulin-2 and its associated ECM protein binding partners may lead to development of novel treatment strategies for KS disease. Citation Information: Cancer Epidemiol Biomarkers Prev 2010;19(10 Suppl):B53.
Cytomegalovirus (CMV) infection is nonpermissive or persistent in many lymphoid and myeloid cell types but can be activated in differentiated macrophages. We have shown elsewhere that both the major immediate-early gene (MIE) and lytic cycle infectious progeny virus expression can be induced in otherwise nonpermissive monocyte-like U-937 cell cultures infected with either human CMV (HCMV) or simian CMV (SCMV) by treatment with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Two multicopy basal enhancer motifs within the SCMV MIE enhancer, namely, 11 copies of the 16-bp cyclic AMP response element (CRE) and 3 copies of novel 17-bp serum response factor (SRF) binding sites referred to as the SNE (SRF/NFkappaB-like element), as well as four classical NFkappaB sites within the HCMV version, contribute to TPA responsiveness in transient assays in monocyte and T-cell types. The SCMV SNE sites contain potential overlapping core recognition binding motifs for SRF, Rel/NFkappaB, ETS, and YY1 class transcription factors but fail to respond to either serum or tumor necrosis factor alpha. Therefore, to evaluate the mechanism of TPA responsiveness of the SNE motifs and of a related 16-bp SEE (SRF/ETS element) motif found in the HCMV and chimpanzee CMV MIE enhancers, we have examined the functional responses and protein binding properties of multimerized wild-type and mutant elements added upstream to the SCMV MIE or simian virus 40 minimal promoter regions in the U-937, K-562, HL-60, THP-1, and Jurkat cell lines. Unlike classical NFkappaB sites, neither the SNE nor the SEE motif responded to phosphatase inhibition by okadaic acid. However, the TPA responsiveness of both CMV elements proved to involve synergistic interactions between the core SRF binding site (CCATATATGG) and the adjacent inverted ETS binding motifs (TTCC), which correlated directly with formation of a bound tripartite complex containing both the cellular SRF and ELK-1 proteins. This protein complex was more abundant in U-937, K-562, and HeLa cell extracts than in Raji, HF, BALB/c 3T3, or HL-60 cells, but the binding activity was altered only twofold after TPA treatment. A 40-fold stimulation of chloramphenicol acetyltransferase activity mediated by four tandem repeats of the SNE could be induced within 2 h (and up to 250-fold within 6 h) after addition of TPA in DNA-transfected U-937 cells, indicating that the stimulation appeared likely to be a true protein kinase C-mediated signal transduction event rather than a differentiation response. Slight differences in the sequence of the core SRF binding site compared with that of the classical c-Fos promoter serum response element, together with differences in the spacing between the SRF and ETS motifs, appear to account for the inability of the SCMV SNEs to respond to serum induction.
Objective: To ensure clinicians can rely on point-of-care testing results, we assessed agreement between point-of-care tests for creatinine, urea, sodium, potassium, calcium, Hb, INR, CRP and subsequent corresponding laboratory tests. Participants: Community-dwelling adults referred to a community-based acute ambulatory care unit. Interventions: The Abbott i-STATTM (Hb, clinical chemistry, INR) and the AfinionTM Analyser (CRP) and corresponding laboratory analyses. Outcomes: Agreement (Bland-Altman) and bias (Passing-Bablok regression). Results: Among 462 adults we found an absolute mean difference between point-of-care and central laboratory analyses of 6.4g/L (95%LOA -7.9 to +20.6) for haemoglobin, -0.5mmol/L (95%LOA -4.5 to +3.5) for sodium, 0.2mmol/L (95%LOA -0.6 to +0.9) for potassium, 0.0mmol/L (95%LOA -0.3 to +0.3) for calcium, 9.0 μmol/L (95%LOA -18.5 to +36.4) for creatinine, 0.0mmol/L (95%LOA -2.7 to +2.6) for urea, -0.2 (95%LOA -2.4 to +2.0) for INR, -5.0 mg/L (95%LOA -24.4 to +14.4) for CRP. Conclusions: There was acceptable agreement and bias for these analytes, except for haemoglobin and creatinine.
Two small fragments of a novel human gammaherpesvirus genome known as Kaposi's sarcoma (KS)-associated herpesvirus or human herpesvirus 8 (HHV-8) have been shown to be present in virtually all AIDS and non-AIDS KS lesions, as well as in body cavity-based lymphomas (BCBL) and in multicentric Castleman's disease. We have extended those studies by identifying and sequencing a third fragment of HHV-8 DNA encoding a viral thymidylate synthetase (TS) gene. Use of this viral TS fragment as a probe led to the identification and mapping of a cluster of overlapping phage lambda clones from a BCBL tumor DNA genomic library that spanned 48 kb on the left-hand side of the HHV-8 genome between the equivalents of open reading frame 6 (ORF6) and ORF31 of herpesvirus saimiri (HVS). DNA sequencing of a 17-kb segment encompassing a gammaherpesvirus divergent locus (DL-B) between ORF11 and ORF17 revealed the presence of nine viral ORFs with predicted gene products related to cellular proteins. These include the complete TS gene and a dihydrofolate reductase (DHFR) gene, four novel cytokine genes (encoding viral interleukin-6, viral MIP-1A, viral MIP-1B, and BCK) that have not previously been found to be encoded by a virus, and a bcl-2 homolog. This region in HHV-8 also contains the T1.1 abundant lytic cycle nuclear RNA gene and encompasses two genes (or exons) encoding proteins with C4HC3 zinc finger domains of the PHD/leukemia-associated protein subtype. The latter are related to the spliced immediate-early IE1 protein of the gamma-2 class herpesvirus bovine herpesvirus type 4 and a similar motif found in HVS ORF12. Although genes for TS and DHFR enzymes are also encoded by HVS (ORF70 and ORF2), both occur at different genomic loci than in HHV-8, and the HHV-8 DHFR protein is much farther diverged from human DHFR than is the HVS version, implying that they were probably acquired as host cell cDNAs by independent evolutionary events. Transcripts from the IE1-A, IE1-B, DHFR, and MIP-1B genes were all detected by Northern blot hybridization analysis in a BCBL cell line at 12 h after induction with butyrate but were not present before induction, indicating that these are all primarily lytic cycle genes. We conclude that the DL-B locus of gammaherpesviruses displays considerably more variability that previously appreciated and that expression of many of these genes is likely to have important implications for HHV-8 biology and therapy.
Objectives Out-of-hours (OOH) primary care services are contacted in the last 4 weeks of life by nearly 30% of all patients who die, but OOH palliative prescribing remains poorly understood. Our understanding of prescribing demand has previously been limited by difficulties identifying palliative patients seen OOH. This study examines the volume and type of prescriptions issued by OOH services at the end of life. Methods A retrospective cohort study was performed by linking a database of Oxfordshire OOH service contacts over a year with national mortality data, identifying patients who died within 30 days of OOH contact. Demographic, service and prescribing data were analysed. Results A prescription is issued at 14.2% of contacts in the 30 days prior to death, compared with 29.9% of other contacts. The most common prescriptions were antibiotics (22.2%) and strong opioids (19%). 41.8% of prescriptions are for subcutaneously administered medication. Patients who were prescribed a syringe driver medication made twice as many OOH contacts in the 30 days prior to death compared with those who were not. Conclusion Absolute and relative prescribing rates are low in the 30 days prior to death. Further research is required to understand what occurs at these non-prescribing end of life contacts to inform how OOH provision can best meet the needs of dying patients. Overall, relatively few patients are prescribed strong opioids or syringe drivers. When a syringe driver medication is prescribed this may help identify patients likely to be in need of further support from the service.
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