Search

BackgroundRespiratory metagenomics (RMg) needs evaluation in a pilot service setting to determine utility and inform implementation into clinical practice.MethodsFeasibility, performance and clinical impacts on antimicrobial prescribing and infection control were recorded during a pilot RMg service for patients with suspected lower respiratory tract infection (LRTI) on two general and one specialist respiratory intensive care units (ICU) at Guy’s & St Thomas NHS foundation Trust, London.ResultsRMg was performed on 128 samples from 87 patients during the first 15-weeks providing same-day results for 110 samples (86%) with median turnaround time of 6.7hrs (IQR 6.1-7.5 hrs). RMg was 92% sensitive and 82% specific for clinically-relevant pathogens compared with routine testing. 48% of RMg results informed antimicrobial prescribing changes (22% escalation; 26% de-escalation) with escalation based on speciation in 20/24 cases and detection of acquired-resistance genes in 4/24 cases. Fastidious or unexpected organisms were reported in 21 samples including anaerobes (n=12),Mycobacterium tuberculosis, Tropheryma whipplei, cytomegalovirus andLegionella pneumophilaST1326, which was subsequently isolated from the bed-side water outlet. Application to consecutive severe community-acquired LRTI cases identifiedStaphylococcus aureus(two withSCCmecand three withlukF/S virulence determinants),Streptococcus pyogenes(emm1-M1uk clone),S. dysgalactiaesubspecies equisimilis (STG62647A) andAspergillus fumigatuswith multiple treatments and public-health impacts.ConclusionsRMg provides frequent diverse benefits for treatment, infection control and public health. The combination of rapid comprehensive results, alongside revealing and characterising a hidden burden of infections makes the case for expediting routine service implementation.

COG-UK is supported by funding from the MRC part of UK Research & Innovation, the National Institute of Health Research (Grant code MC_PC_19027), and Genome Research Limited, operating as the Welcome Sanger Institute. Background The first epidemic wave of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Scotland resulted in high case numbers and mortality in care homes. In Lothian, over one-third of care homes reported an outbreak, while there was limited testing of hospital patients discharged to care homes. Aim To investigate patients discharged from hospitals as a source of SARS-CoV-2 introduction into care homes during the first epidemic wave. Methods A clinical review was performed for all patients discharges from hospitals to care homes from 1st March 2020 to 31st May 2020. Episodes were ruled out based on coronavirus disease 2019 (COVID-19) test history, clinical assessment at discharge, whole-genome sequencing (WGS) data and an infectious period of 14 days. Clinical samples were processed for WGS, and consensus genomes generated were used for analysis using Cluster Investigation and Virus Epidemiological Tool software. Patient timelines were obtained using electronic hospital records. Findings In total, 787 patients discharged from hospitals to care homes were identified. Of these, 776 (99%) were ruled out for subsequent introduction of SARS-CoV-2 into care homes. However, for 10 episodes, the results were inconclusive as there was low genomic diversity in consensus genomes or no sequencing data were available. Only one discharge episode had a genomic, time and location link to positive cases during hospital admission, leading to 10 positive cases in their care home. Conclusion The majority of patients discharged from hospitals were ruled out for introduction of SARS-CoV-2 into care homes, highlighting the importance of screening all new admissions when faced with a novel emerging virus and no available vaccine. Publisher PDF

Background: Thyroid gland is one the important organs, which play a wide and vital physiological role in the body. Thyroid diseases are quite common. The incidence of thyroid diseases depends mainly depending upon iron deficiency status. Fine-needle aspiration cytology (FNAC) plays a vital role in evaluating patients with thyroid nodules. Surgical excision and histopathological evaluation are crucial to establish the diagnosis. Aims and Objectives: This study mainly aims to classify the cytomorphology of palpable thyroid lesions by FNAC and correlate the results with histopathology. This study also aims to determine the spectrum of histopathological diagnosis in thyroid surgeries. Materials and Methods: The present study was carried out at cytopathology and histopathology section of pathology department of GMERS Medical College, Valsad. In present study, we analysed 276 cases of palpable thyroid nodule underwent FNAC. Out of them 152 cases underwent surgical procedure like lobectomy or partial or total thyroidectomy Results: Out of these 276 patient, females were 218 and males were 58 with F:M ratio of 3.76:1. Out of total 276 patients, 81 (29.3%) were between 31-40 years followed by 75 (27.2%) were between 21-30 years and 237 (85.9%) were fall into category-II. Out of these 152 patients, 88 (57.9%) having colloid goitre and 79 (89.7%) were confirmed histologically. Conclusion: FNAC is an invaluable tool in management of thyroid lesions. It is a safe, simple and cost effective procedure and can be performed on outdoor patients. If the cytology report is malignant, surgery is the recommended procedure; for suspicious lesions, repeat aspiration is required and for benign lesions no further immediate diagnostic studies are required. Keywords: Endocrine, Thyroid, Histopathology correlation, Inflammatory, Neoplastic.

Background: \ud The SARS-CoV-2 delta (B.1.617.2) variant was first detected in England in March, 2021. It has since rapidly become the predominant lineage, owing to high transmissibility. It is suspected that the delta variant is associated with more severe disease than the previously dominant alpha (B.1.1.7) variant. We aimed to characterise the severity of the delta variant compared with the alpha variant by determining the relative risk of hospital attendance outcomes.\ud \ud Methods: \ud This cohort study was done among all patients with COVID-19 in England between March 29 and May 23, 2021, who were identified as being infected with either the alpha or delta SARS-CoV-2 variant through whole-genome sequencing. Individual-level data on these patients were linked to routine health-care datasets on vaccination, emergency care attendance, hospital admission, and mortality (data from Public Health England's Second Generation Surveillance System and COVID-19-associated deaths dataset; the National Immunisation Management System; and NHS Digital Secondary Uses Services and Emergency Care Data Set). The risk for hospital admission and emergency care attendance were compared between patients with sequencing-confirmed delta and alpha variants for the whole cohort and by vaccination status subgroups. Stratified Cox regression was used to adjust for age, sex, ethnicity, deprivation, recent international travel, area of residence, calendar week, and vaccination status.\ud \ud Findings: \ud Individual-level data on 43 338 COVID-19-positive patients (8682 with the delta variant, 34 656 with the alpha variant; median age 31 years [IQR 17–43]) were included in our analysis. 196 (2·3%) patients with the delta variant versus 764 (2·2%) patients with the alpha variant were admitted to hospital within 14 days after the specimen was taken (adjusted hazard ratio [HR] 2·26 [95% CI 1·32–3·89]). 498 (5·7%) patients with the delta variant versus 1448 (4·2%) patients with the alpha variant were admitted to hospital or attended emergency care within 14 days (adjusted HR 1·45 [1·08–1·95]). Most patients were unvaccinated (32 078 [74·0%] across both groups). The HRs for vaccinated patients with the delta variant versus the alpha variant (adjusted HR for hospital admission 1·94 [95% CI 0·47–8·05] and for hospital admission or emergency care attendance 1·58 [0·69–3·61]) were similar to the HRs for unvaccinated patients (2·32 [1·29–4·16] and 1·43 [1·04–1·97]; p=0·82 for both) but the precision for the vaccinated subgroup was low.\ud \ud Interpretation: \ud This large national study found a higher hospital admission or emergency care attendance risk for patients with COVID-19 infected with the delta variant compared with the alpha variant. Results suggest that outbreaks of the delta variant in unvaccinated populations might lead to a greater burden on health-care services than the alpha variant.\ud \ud Funding: \ud Medical Research Council; UK Research and Innovation; Department of Health and Social Care; and National Institute for Health Research.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections were rising during early summer 2021 in many countries as a result of the Delta variant. We assessed reverse transcription polymerase chain reaction swab positivity in the Real-time Assessment of Community Transmission–1 (REACT-1) study in England. During June and July 2021, we observed sustained exponential growth with an average doubling time of 25 days, driven by complete replacement of the Alpha variant by Delta and by high prevalence at younger, less-vaccinated ages. Prevalence among unvaccinated people [1.21% (95% credible interval 1.03%, 1.41%)] was three times that among double-vaccinated people [0.40% (95% credible interval 0.34%, 0.48%)]. However, after adjusting for age and other variables, vaccine effectiveness for double-vaccinated people was estimated at between ~50% and ~60% during this period in England. Increased social mixing in the presence of Delta had the potential to generate sustained growth in infections, even at high levels of vaccination.

Background Clinical metagenomics (CMg) has the potential to be translated from a research tool into routine service to improve antimicrobial treatment and infection control decisions. The SARS-CoV-2 pandemic provides added impetus to realise these benefits, given the increased risk of secondary infection and nosocomial transmission of multi-drug-resistant (MDR) pathogens linked with the expansion of critical care capacity. Methods CMg using nanopore sequencing was evaluated in a proof-of-concept study on 43 respiratory samples from 34 intubated patients across seven intensive care units (ICUs) over a 9-week period during the first COVID-19 pandemic wave. Results An 8-h CMg workflow was 92% sensitive (95% CI, 75–99%) and 82% specific (95% CI, 57–96%) for bacterial identification based on culture-positive and culture-negative samples, respectively. CMg sequencing reported the presence or absence of β-lactam-resistant genes carried by Enterobacterales that would modify the initial guideline-recommended antibiotics in every case. CMg was also 100% concordant with quantitative PCR for detecting Aspergillus fumigatus from 4 positive and 39 negative samples. Molecular typing using 24-h sequencing data identified an MDR-K. pneumoniae ST307 outbreak involving 4 patients and an MDR-C. striatum outbreak involving 14 patients across three ICUs. Conclusion CMg testing provides accurate pathogen detection and antibiotic resistance prediction in a same-day laboratory workflow, with assembled genomes available the next day for genomic surveillance. The provision of this technology in a service setting could fundamentally change the multi-disciplinary team approach to managing ICU infections. The potential to improve the initial targeted treatment and rapidly detect unsuspected outbreaks of MDR-pathogens justifies further expedited clinical assessment of CMg.

Fine needle aspiration cytology (FNAC) is a relatively simpler tool as compared to histopathology for the evaluation of the cytomorphology of skin lesions. Fifty consecutive leprosy patients diagnosed and classified according to Ridley Jopling scale1 were taken to study the cytomorphology of leprosy lesions in fine needle aspirates and correlate it with their histopathology by a skin biopsy from the same site for histopathological examination after H/E staining. Borderline leprosy patients with Type I reaction showed significantly more numbers of giant cells, collagen and elastin in their smears as compared to those without reaction. The smears were more heavily cellular with fragmented collagen and elastin along with significant increase in neutrophils in patients with Type II reaction while foamy macrophages with fatty background were common in non-reactional lepromatous leprosy patients. A complete correlation between histopathological and cytomorphological findings was observed in 79.4% of cases.

Background: Breast cancer is the most common cancer among women in the western world and second most in India after carcinoma cervix. The most common presentation of breast disease is a palpable mass with or without inflammatory lesion, nipple discharge and imaging abnormalities. Fine Needle Aspiration Cytology (FNAC) is a popular diagnostic technique as it is highly sensitive, specific, rapid, easy to perform, cost effective and can be carried out at OPD level.in the first visit without the need of anaesthesia. This technique is commonly used in combination with physical examination and mammography, so called triple-test diagnostic triad. Triple assessment of breast mass had decreased the false negative rate to less than 1%.Aims and Objectives: The aim of this study was to find out the common causes of breast lump, to find out sensitivity and specificity of FNAC of breast lump and also to calculate statistical data like sensitivity, specificity, positive predictive value, negative predictive value and efficiency of FNAC procedure.Materials and Methods: This study comprised of 180 cases and carried out at histopathology section of pathology department of GMERS Medical College, Valsad, a tertiary care centre in Valsad district of Gujarat state, India. The clinical case history and radiological details of the patient was recorded which includes detail history of pain, nipple discharge, ulceration of nipple and duration of lesion. Results: Out of total 180 cases, 126 (70.0%) were benign and 54 (30.0%) were malignant. Out of total 180 cases, 95 (52.8%) were located in right sided breast, 82 (45.5%) were located in left sided breast and 03 (2.4%) were located bilaterally. Out of 121 histologically diagnosed benign cases, 101 (83.5%) were fibroadenoma. Out of total 59 histologically diagnosed malignant cases, 43 (72.9%) were intraductal carcinoma. On FNAC of breast lumps, 70% (n=126) were categorized as benign, 30% (n=54) as malignant. Subsequent histopathology of breast lumps revealed 67.2% (n=121) benign lesions and 32.8% (n=59) malignant lesions.Conclusion: The FNAC is an essential and important test in diagnosing breast lump. It is patient-friendly, easy to perform, more reliable, repeatable, cost effective and a simple and safe test to diagnose patient presenting with lump breast. FNAC can suggest further investigation without delay if carcinoma could be diagnosed. It also help the clinicians for early diagnosis and specific management thus reducing morbidity and mortality.

Summary Background Lateral flow devices (LFDs) for rapid antigen testing are set to become a cornerstone of SARS-CoV-2 mass community testing, although their reduced sensitivity compared with PCR has raised questions of how well they identify infectious cases. Understanding their capabilities and limitations is, therefore, essential for successful implementation. We evaluated six commercial LFDs and assessed their correlation with infectious virus culture and PCR cycle threshold (Ct) values. Methods In a single-centre, laboratory evaluation study, we did a head-to-head comparison of six LFDs commercially available in the UK: Innova Rapid SARS-CoV-2 Antigen Test, Spring Healthcare SARS-CoV-2 Antigen Rapid Test Cassette, E25Bio Rapid Diagnostic Test, Encode SARS-CoV-2 Antigen Rapid Test Device, SureScreen COVID-19 Rapid Antigen Test Cassette, and SureScreen COVID-19 Rapid Fluorescence Antigen Test. We estimated the specificities and sensitivities of the LFDs using stored naso-oropharyngeal swabs collected at St Thomas' Hospital (London, UK) for routine diagnostic SARS-CoV-2 testing by real-time RT-PCR (RT-rtPCR). Swabs were from inpatients and outpatients from all departments of St Thomas' Hospital, and from health-care staff (all departments) and their household contacts. SARS-CoV-2-negative swabs from the same population (confirmed by RT-rtPCR) were used for comparative specificity determinations. All samples were collected between March 23 and Oct 27, 2020. We determined the limit of detection (LOD) for each test using viral plaque-forming units (PFUs) and viral RNA copy numbers of laboratory-grown SARS-CoV-2. Additionally, LFDs were selected to assess the correlation of antigen test result with RT-rtPCR Ct values and positive viral culture in Vero E6 cells. This analysis included longitudinal swabs from five infected inpatients with varying disease severities. Furthermore, the sensitivities of available LFDs were assessed in swabs (n=23; collected from Dec 4, 2020, to Jan 12, 2021) confirmed to be positive (RT-rtPCR and whole-genome sequencing) for the B.1.1.7 variant, which was the dominant genotype in the UK at the time of study completion. Findings All LFDs showed high specificity (≥98·0%), except for the E25Bio test (86·0% [95% CI 77·9–99·9]), and most tests reliably detected 50 PFU/test (equivalent SARS-CoV-2 N gene Ct value of 23·7, or RNA copy number of 3 × 106/mL). Sensitivities of the LFDs on clinical samples ranged from 65·0% (55·2–73·6) to 89·0% (81·4–93·8). These sensitivities increased to greater than 90% for samples with Ct values of lower than 25 for all tests except the SureScreen fluorescence (SureScreen-F) test. Positive virus culture was identified in 57 (40·4%) of 141 samples; 54 (94·7%) of the positive cultures were from swabs with Ct values lower than 25. Among the three LFDs selected for detailed comparisons (the tests with highest sensitivity [Innova], highest specificity [Encode], and alternative technology [SureScreen-F]), sensitivity of the LFDs increased to at least 94·7% when only including samples with detected viral growth. Longitudinal studies of RT-rtPCR-positive samples (tested with Innova, Encode, and both SureScreen-F and the SureScreen visual [SureScreen-V] test) showed that most of the tests identified all infectious samples as positive. Test performance (assessed for Innova and SureScreen-V) was not affected when reassessed on swabs positive for the UK variant B.1.1.7. Interpretation In this comprehensive comparison of antigen LFDs and virus infectivity, we found a clear relationship between Ct values, quantitative culture of infectious virus, and antigen LFD positivity in clinical samples. Our data support regular testing of target groups with LFDs to supplement the current PCR testing capacity, which would help to rapidly identify infected individuals in situations in which they would otherwise go undetected. Funding King's Together Rapid COVID-19, Medical Research Council, Wellcome Trust, Huo Family Foundation, UK Department of Health, National Institute for Health Research Comprehensive Biomedical Research Centre.

BackgroundRapid antigen lateral flow devices (LFDs) are set to become a cornerstone of SARS-CoV-2 mass community testing. However, their reduced sensitivity compared to PCR has raised questions of how well they identify infectious cases. Understanding their capabilities and limitations is therefore essential for successful implementation. To address this, we evaluated six commercial LFDs on the same collection of clinical samples and assessed their correlation with infectious virus culture and cycle threshold (Ct) values.MethodsA head-to-head comparison of specificities and sensitivities was performed on six commercial rapid antigen tests using combined nasal/oropharyngeal swabs, and their limits of detection determined using viral plaque forming units (PFU). Three of the LFDs were selected for a further study, correlating antigen test result with RT-PCR Ct values and positive viral culture in Vero-E6 cells. This included sequential swabs and matched serum samples obtained from four infected individuals with varying disease severities. Detection of antibodies was performed using an IgG/IgM Rapid Test Cassette, and neutralising antibodies by infectious virus assay. Finally, the sensitivities of selected rapid antigen LFTs were assessed in swabs with confirmed B.1.1.7 variant, currently the dominant genotype in the UK.FindingsMost of the rapid antigen LFDs showed a high specificity (>98%), and accurately detected 50 PFU/test (equivalent N1 Ct of 23.7 or RNA copy number of 3×106/ml). Sensitivities of the LFDs performed on clinical samples ranged from 65 to 89%. These sensitivities increased in most tests to over 90% for samples with Cts lower than 25. Positive virus culture was achieved for 57 out of 141 samples, with 80% of the positive cultures from swabs with Cts lower than 23. Importantly, sensitivity of the LFDs increased to over 95% when compared with the detection of infectious virus alone, irrespective of Ct. Longitudinal studies of PCR-positive samples showed that most of the tests identified all infectious samples as positive, but differences in test sensitivities can lead to missed cases in the absence of repeated testing. Finally, test performance was not impacted when re-assessed against swabs positive for the dominant UK variant B.1.1.7.InterpretationIn this comprehensive comparison of antigen LFD and virus infectivity, we demonstrate a clear relationship between Ct values, quantitative culture of infectious virus and antigen LFD positivity in clinical samples. Our data support regular testing of target groups using LFDs to supplement the current PCR testing capacity, to rapidly identify infected individuals in situations where they would otherwise go undetected.FundingKing’s Together Rapid COVID-19, Medical Research Council, Wellcome Trust, Huo Family Foundation.

BackgroundClinical metagenomics (CMg) is being evaluated for translation from a research tool into routine diagnostic service, but its potential to significantly improve management of acutely unwell patients has not been demonstrated. The SARS-CoV-2 pandemic provides impetus to determine that benefit given increased risk of secondary infection and nosocomial transmission by multi-drug resistant (MDR) pathogens linked with expansion of critical care capacity.MethodsProspective evaluation of CMg using nanopore sequencing was performed on 43 respiratory samples over 14 weeks from a cohort of 274 intubated patients across seven COVID-19 intensive care units.ResultsBacteria or fungi were cultured from 200 (73%) patients, with a predominance of Klebsiella spp. (31%) and C. striatum (7%) amongst other common respiratory pathogens. An 8 hour CMg workflow was 93% sensitive and 81% specific for bacterial identification compared to culture, and reported presence or absence of β-lactam resistance genes carried by Enterobacterales that would modify initial guideline-recommended antibiotics in every case. CMg was also 100% concordant with quantitative PCR for detecting Aspergillus fumigatus (4 positive and 39 negative samples). Single nucleotide polymorphism (SNP)-typing using 24 hour sequence data identified an MDR-K. pneumoniae ST307 outbreak involving 4 patients and an MDR-C. striatum outbreak potentially involving 14 patients across three ICUs.ConclusionCMg testing for ICU patients provides same-day pathogen detection and antibiotic resistance prediction that significantly improves initial treatment of nosocomial pneumonia and rapidly detects unsuspected outbreaks of MDR-pathogens.

Antibody responses to SARS-CoV-2 can be detected in most infected individuals 10–15 d after the onset of COVID-19 symptoms. However, due to the recent emergence of SARS-CoV-2 in the human population, it is not known how long antibody responses will be maintained or whether they will provide protection from reinfection. Using sequential serum samples collected up to 94 d post onset of symptoms (POS) from 65 individuals with real-time quantitative PCR-confirmed SARS-CoV-2 infection, we show seroconversion (immunoglobulin (Ig)M, IgA, IgG) in >95% of cases and neutralizing antibody responses when sampled beyond 8 d POS. We show that the kinetics of the neutralizing antibody response is typical of an acute viral infection, with declining neutralizing antibody titres observed after an initial peak, and that the magnitude of this peak is dependent on disease severity. Although some individuals with high peak infective dose (ID50 > 10,000) maintained neutralizing antibody titres >1,000 at >60 d POS, some with lower peak ID50 had neutralizing antibody titres approaching baseline within the follow-up period. A similar decline in neutralizing antibody titres was observed in a cohort of 31 seropositive healthcare workers. The present study has important implications when considering widespread serological testing and antibody protection against reinfection with SARS-CoV-2, and may suggest that vaccine boosters are required to provide long-lasting protection.

Antibody (Ab) responses to SARS-CoV-2 can be detected in most infected individuals 10-15 days following the onset of COVID-19 symptoms. However, due to the recent emergence of this virus in the human population it is not yet known how long these Ab responses will be maintained or whether they will provide protection from re-infection. Using sequential serum samples collected up to 94 days post onset of symptoms (POS) from 65 RT-qPCR confirmed SARS-CoV-2-infected individuals, we show seroconversion in >95% of cases and neutralizing antibody (nAb) responses when sampled beyond 8 days POS. We demonstrate that the magnitude of the nAb response is dependent upon the disease severity, but this does not affect the kinetics of the nAb response. Declining nAb titres were observed during the follow up period. Whilst some individuals with high peak ID50 (>10,000) maintained titres >1,000 at >60 days POS, some with lower peak ID50 had titres approaching baseline within the follow up period. A similar decline in nAb titres was also observed in a cohort of seropositive healthcare workers from Guy’s and St Thomas’ Hospitals. We suggest that this transient nAb response is a feature shared by both a SARS-CoV-2 infection that causes low disease severity and the circulating seasonal coronaviruses that are associated with common colds. This study has important implications when considering widespread serological testing, Ab protection against re-infection with SARS-CoV-2 and the durability of vaccine protection.

Background: Ovaries are complex intra-pelvic organs of the female reproductive system. Ovarian cancer accounts for 3% of all cancers in females and is the fifth most common cause of cancer death in women. Early menarche, late menopause, nulliparity and high socioeconomic status are associated with an increased risk for ovarian neoplasms. Histo-pathological diagnosis remains the mainstay to differentiate neoplastic lesions from non-neoplastic lesions. Aims and objectives: This study aims to analyze the view of ovarian tumors with respect to clinical presentation, gross and microscopic characteristics and also to study the frequency and histopathological patterns of ovarian tumours. Materials and Methods: This study comprised of 100 cases of ovarian tumours received in the Department of Pathology, GMERS Medical College, Gandhinagar were analysed. Their gross features, microscopic findings were analysed in detail. Ovarian tumours were divided into benign and malignant categories and their further sub typing were done according to WHO Classification. Results: Out of total 100 cases, 76 were benign and 24 were malignant. Out of 100 cases, 62% were between 21-40 years of age followed by 28% were between 41-60 years of age. Most common presenting symptom was pain in abdomen followed by lump in abdomen and heavy menstruation. Out of total 100 cases, 70% were surface epithelial tumours, 24% were germ cell tumours and 6% were sex cord stromal tumours. Conclusion: To conclude we recommend microscopic histopathological examination of every ovarian mass in order to assess the importance of pathological grading and staging and they must be classified correctly so that the patient can be provided with appropriate treatment and prognosis.

There is a worldwide shortage of reagents to perform detection of SARS-2. Many clinical diagnostic laboratories rely on commercial platforms that provide integrated end-to-end solutions. While this provides established robust pipelines, there is a clear bottleneck in the supply of reagents given the current situation of extraordinary high demand. Some laboratories resort to implementing kit-free handling procedures, but many other small laboratories will not have the capacity to develop those and/or will perform manual handling of their samples. In order to provide multiple workflows for SARS-CoV-2 nucleic acid detection we compared several commercially available RNA extraction methods: QIAamp Viral RNA Mini Kit (QIAgen), the recently developed RNAdvance Blood (Beckman) and Mag-Bind Viral DNA/RNA 96 Kit (Omega Bio-tek). We also compared different 1-step RT-qPCR Master Mix brands: TaqMan™ Fast Virus 1-Step Master Mix (ThermoFisher Scientific), qPCRBIO Probe 1-Step Go Lo-ROX (PCR Biosystems) and Luna® Universal Probe One-Step RT-qPCR Kit (NEB). We used the Centre for Disease Control (CDC) recommended primers that detect two regions of the viral N gene as well as those that detect the RdRP gene region as per Public Health England (PHE) guidelines (Charite/WHO/PHE). Our data show that the RNA extraction methods provide similar results. Amongst the qPCR reagents tested, TaqMan™ Fast Virus 1-Step Master Mix and Luna® Universal Probe One-Step RT-qPCR Kit proved most sensitive. The N1 and N2 primer-probes provide a more reliable detection than the RdRP-SARSr primer-probe set, particularly in samples with low viral titres. Importantly, we have implemented a protocol using heat inactivation and demonstrate that it has minimal impact on the sensitivity of the qPCR in clinical samples - potentially making SARS-CoV-2 testing portable to settings that do not have CL-3 facilities.

Objective To examine whether accounting for a woman’s age and body mass index (BMI) would improve the ability of antimullerian hormone (AMH) to distinguish between women with (cases) and without (controls) polycystic ovarian syndrome (PCOS). Design An opportunistic case-control dataset of reproductive age women having evaluations for PCOS as defined by National Institutes of Health criteria. Setting Two medical centers in the United States enrolled women. Serum samples were analyzed for relevant analytes. Patients Women were between 18 and 39 years of age when samples and clinical information were collected. Residual samples had been stored for 2−17 years. AMH was measured via immunoassay. Interventions None; this was an observational study. Main outcome measures Detection and false-positive rates for PCOS were computed for AMH results expressed as multiples of the median (MoM) both before and after adjustment for the woman’s age and BMI. Results Using unadjusted AMH MoM results, 168 cases (78%) cases were at or beyond the 90th centile of controls (2.47 MoM). After accounting for each woman’s age and BMI, 188 (87%) of those women were beyond the 90th centile of controls (2.20 MoM), a significant increase (P = .015). The adjusted AMH MoM levels fitted logarithmic normal distributions well (mean, standard deviation for controls and cases of 0.0000, 0.2765 and 0.6884, 0.2874, respectively) and this allowed for computation of patient-specific PCOS risks. Conclusions Accounting for the woman’s age and BMI resulted in significantly higher AMH-based detection rates for PCOS at a 10% false-positive rate, and patient-specific PCOS risks could be computed.

High-throughput screening (HTS) is a widespread method in early drug discovery for identifying promising chemical matter that modulates a target or phenotype of interest. Because HTS campaigns involve screening millions of compounds, it is often desirable to initiate screening with a subset of the full collection. Subsequently, virtual screening methods prioritize likely active compounds in the remaining collection in an iterative process. With this approach, orthogonal virtual screening methods are often applied, necessitating the prioritization of hits from different approaches. Here, we introduce a novel method of fusing these prioritizations and benchmark it prospectively on 17 screening campaigns using virtual screening methods in three descriptor spaces. We found that the fusion approach retrieves 15% to 65% more active chemical series than any single machine-learning method and that appropriately weighting contributions of similarity and machine-learning scoring techniques can increase enrichment by 1% to 19%. We also use fusion scoring to evaluate the tradeoff between screening more chemical matter initially in lieu of replicate samples to prevent false-positives and find that the former option leads to the retrieval of more active chemical series. These results represent guidelines that can increase the rate of identification of promising active compounds in future iterative screens.

An evaluation of a rapid portable gold-nanotechnology measuring SARS-CoV-2 IgM, IgA and IgG antibody concentrations against spike 1 (S1), spike 2 (S) and nucleocapsid (N) was conducted using serum samples from 74 patients tested for SARS-CoV-2 RNA on admission to hospital, and 47 historical control patients from March 2019. 59 patients were RNA(+) and 15 were RNA(-). A serum (±) classification was derived for all three antigens and a quantitative serological profile was obtained. Serum(+) was identified in 30% (95% CI 11-48) of initially RNA(-) patients, in 36% (95% CI 17-54) of RNA(+) patients before 10 days, 77% (95% CI 67-87) between 10 and 20 days and 95% (95% CI 86-100) after 21 days. The patient-level diagnostic accuracy relative to RNA(±) after 10 days displayed 88% sensitivity (95% CI 75-95) and 75% specificity (95% CI 22-99), although specificity compared with historical controls was 100% (95%CI 91-100). This study provides robust support for further evaluation and validation of this novel technology in a clinical setting and highlights challenges inherent in assessment of serological tests for an emerging disease such as COVID-19.

In contrast to human cells, very few HSV-1 genes are known to be spliced, although the same pre-mRNA processing machinery is shared. Here, through global analysis of splice junctions in cells infected with HSV-1 and an HSV-1 mutant virus with deletion of infectious cell culture protein 27 (ICP27), one of two viral immediate early (IE) genes essential for viral replication, we identify hundreds of novel alternative splice junctions mapping to both previously known HSV-1 spliced genes and previously unknown spliced genes, the majority of which alter the coding potential of viral genes. Quantitative and qualitative splicing efficiency analysis of these novel alternatively spliced genes based on RNA-Seq and RT-PCR reveals that splicing at these novel splice sites is efficient only when ICP27 is absent; while in wildtype HSV-1 infected cells, the splicing of these novel splice junctions is largely silenced in a gene/sequence specific manner, suggesting that ICP27 not only promotes accumulation of ICP27 targeted transcripts but also ensures correctness of the functional coding sequences through inhibition of alternative splicing. Furthermore, ICP27 toggles expression of ICP34.5, the major viral neurovirulence factor, through inhibition of splicing and activation of a proximal polyadenylation signal (PAS) in the newly identified intron, revealing a novel regulatory mechanism for expression of a viral gene. Thus, through the viral IE protein ICP27, HSV-1 co-opts both splicing and polyadenylation machinery to achieve optimal viral gene expression during lytic infection. On the other hand, during latent infection when ICP27 is absent, HSV-1 likely takes advantages of host splicing machinery to restrict expression of randomly activated antigenic viral genes to achieve immune evasion., Author summary Little is known regarding to how HSV, a large DNA virus and known to contain very few spliced genes, escapes host pre-mRNA splicing machinery. Here, by establishing a high throughput splice junction identification platform and quantitative analysis method to assess splicing efficiency based on high throughput data, we find that HSV-1 encodes hundreds of previously unknown alternative splice junctions; however, splicing of these novel spliced genes is largely silenced in wild-type HSV-1 infected cells, explaining why only very few spliced genes have been previously identified in HSV-1. Moreover, ICP27 is required for splicing inhibition and 3’ end formation of ICP34.5, the major viral neurovirulence factor and also the major target of latently expressed viral miRNAs. These findings not only fundamentally change the view of HSV gene structure, but also reveal a mechanism by which HSV employs host splicing and polyadenylation machineries to achieve optimal gene expression during acute infection and may also contribute to immune evasion during latency when ICP27 is not expressed.

Objective: Several studies showed that a substantial decline in the use of co-trimoxazole did not result in a decline in resistance rates among Escherichia coli isolates. Since mathematical models have shown that it may take decades before resistance rates start to decline to relevant levels, we performed a new analysis using more recently collected data. Methods: Data were extracted from Guy’s and St Thomas’ Hospitals Transmission and Antimicrobial Record database which contains microbiological test results from all specimens tested between 2002 and 2014. We selected all blood samples positive for E. coli which were tested for resistance against co-trimoxazole. Prevalence of co-trimoxazole resistance among the tested samples by year was modelled by a Poisson model. Results: Almost all (96%) of E. coli blood isolates were tested for co-trimoxazole resistance. In total, 2,070 E. coli isolates were available for analyses. Resistance to co-trimoxazole fluctuated over the years, but there was no clear increasing or decreasing trend; the annual percentage change in the prevalence of co-trimoxazole resistance was 0.52 (95% confidence interval −0.75% to 1.81%). Including co-trimoxazole or trimethoprim use in the year before the sample was taken did not improve the model. Conclusion: The prevalence of co-trimoxazole resistance among E. coli blood isolates remained high, almost three decades after a substantial decline in co-trimoxazole use. Our results further emphasize the importance of prudent antibiotics use, as antibiotic resistance may not always be easily reversible.

The herpes simplex virus (HSV) infected cell culture polypeptide 27 (ICP27) protein is essential for virus infection of cells. Recent studies suggested that ICP27 inhibits splicing in a gene-specific manner via an unknown mechanism. Here, RNA-sequencing revealed that ICP27 not only inhibits splicing of certain introns in

Despite the long-standing observation that herpes simplex virus (HSV) latency-associated transcript (LAT) promoter deletion viruses show impaired recurrence phenotypes in relevant animal models, the mechanism by which these sequences exert this phenotypic effect is unknown. We constructed and evaluated four mutant HSV-2 isolates with targeted mutations in the LAT promoter and LAT-associated microRNAs (miRNAs) affecting (i) the LAT TATA box; (ii) the LAT ICP4-binding site; (iii) miRNA I (miR-I) and miR-II (miR-I/II), which both target ICP34.5; and (iv) miR-III, which targets ICP0. While the LAT TATA box mutant caused milder acute infections than wild-type (WT) virus, there was no difference in the recurrence phenotype between these viruses. LAT and miRNA expression during latency was not impaired by this mutation, suggesting that other promoter elements may be more important for latent HSV-2 LAT expression. Mutation of the LAT ICP4-binding site also did not cause an in vivo phenotypic difference between mutant and WT viruses. Acute infection and reactivation from latency of the miR-I/II mutant were similar to those of its rescuant, although the acute infection was slightly reduced in severity relative to that caused by the wild-type virus. The miR-III mutant also exhibited WT phenotypes in acute and recurrent phases of infection. While they do not rule out an effect of these elements in human latency or reactivation, these findings do not identify a specific role for LAT or LAT-associated miRNAs in the HSV-2 LAT promoter deletion phenotype in guinea pigs. Thus, other sequences in this region may play a more important role in the long-studied LAT-associated phenotype in animals. IMPORTANCE While it has been known for several decades that specific HSV-1 and HSV-2 sequences near the LAT promoter are required for efficient viral reactivation in animal models, the mechanism is still not known. We constructed four mutant viruses with the goal of identifying critical sequence elements and of specifically testing the hypothesis that microRNAs that are expressed during latency play a role. Determination that specific LAT promoter sequences and miRNA sequences do not influence viral reactivation of HSV-2 helps to narrow down the search for the mechanism by which the virus controls its latency and recurrence phenotype.

BACKGROUND: Staphylococcus aureus bacteraemia is a common cause of severe community-acquired and hospital-acquired infection worldwide. We tested the hypothesis that adjunctive rifampicin would reduce bacteriologically confirmed treatment failure or disease recurrence, or death, by enhancing early S aureus killing, sterilising infected foci and blood faster, and reducing risks of dissemination and metastatic infection.METHODS: In this multicentre, randomised, double-blind, placebo-controlled trial, adults (≥18 years) with S aureus bacteraemia who had received ≤96 h of active antibiotic therapy were recruited from 29 UK hospitals. Patients were randomly assigned (1:1) via a computer-generated sequential randomisation list to receive 2 weeks of adjunctive rifampicin (600 mg or 900 mg per day according to weight, oral or intravenous) versus identical placebo, together with standard antibiotic therapy. Randomisation was stratified by centre. Patients, investigators, and those caring for the patients were masked to group allocation. The primary outcome was time to bacteriologically confirmed treatment failure or disease recurrence, or death (all-cause), from randomisation to 12 weeks, adjudicated by an independent review committee masked to the treatment. Analysis was intention to treat. This trial was registered, number ISRCTN37666216, and is closed to new participants.FINDINGS: Between Dec 10, 2012, and Oct 25, 2016, 758 eligible participants were randomly assigned: 370 to rifampicin and 388 to placebo. 485 (64%) participants had community-acquired S aureus infections, and 132 (17%) had nosocomial S aureus infections. 47 (6%) had meticillin-resistant infections. 301 (40%) participants had an initial deep infection focus. Standard antibiotics were given for 29 (IQR 18-45) days; 619 (82%) participants received flucloxacillin. By week 12, 62 (17%) of participants who received rifampicin versus 71 (18%) who received placebo experienced treatment failure or disease recurrence, or died (absolute risk difference -1·4%, 95% CI -7·0 to 4·3; hazard ratio 0·96, 0·68-1·35, p=0·81). From randomisation to 12 weeks, no evidence of differences in serious (p=0·17) or grade 3-4 (p=0·36) adverse events were observed; however, 63 (17%) participants in the rifampicin group versus 39 (10%) in the placebo group had antibiotic or trial drug-modifying adverse events (p=0·004), and 24 (6%) versus six (2%) had drug interactions (p=0·0005).INTERPRETATION: Adjunctive rifampicin provided no overall benefit over standard antibiotic therapy in adults with S aureus bacteraemia.FUNDING: UK National Institute for Health Research Health Technology Assessment.

The population of the United States is aging. Currently people over the age of 65 years constitute 13% of the general population. By 2050 this number will rise to about 25% of the population. As the population ages, the number of older adults with mental illness will also rise. Available data indicates that there are inadequate numbers of trained geriatric psychiatrists in United States to care for older adults with mental illness. The additional burden for services in the future on an already strained healthcare system can lead to catastrophic failure of the system. International Medical Graduates (IMGs) constitute almost half of the work force of geriatric psychiatrists. The IMGs have had successful career as clinicians, educators, academics and researchers in geriatric psychiatry. In this symposium we will enumerate the unique challenges faced by the IMGs and strategies on enable them to integrate into the mainstream geriatric psychiatry workforce. We will review the roles of IMGs as private practitioners, educators, academicians and as researchers. We will also discuss how organizations like the AAGP can play a greater role in attracting larger number of IMGs to join the geriatric psychiatry workforce and also the AAGP. This will enable greater access to care for the older adults with mental illness and maintain the success of AAGP in being the national organization for geriatric psychiatry clinicians in the United States.

Despite advances in the neurosciences, we continue to struggle with the treatment of neuropsychiatric symptoms in persons with dementia. There are very limited available treatments that are FDA approved. Recent statements and regulations have made the treatment of some neuropsychiatric symptoms of dementia more challenging. Despite the known risks of antipsychotics, APA has issued the statement indicating that there are still specific clinical situations which their use may be clinically appropriate. Conversely, CMS regulations have increased scrutiny of the use of antipsychotic medications in the Long-Term Care setting. The best interest of the patient seems to have been minimized, making it more difficult for us as geriatric psychiatrists to provide appropriate treatment for our patients. Our panel will discuss the following 1. The current data from examining antipsychotic and other psychotropic use among residents in long-term care using a national sample of Medicare data from 2009-2014, specifically focusing on antipsychotics and mood stabilizers. 2. The current CMS regulation of psychotropic medications prescribing with an emphasis on antipsychotics medication in Long-Term Care 3. The current public perception of dementia care 4. How CMS regulations of psychotropic medications affect the treatment of psychiatric patients and dementia care in nursing homes. 5. What we, as geriatric psychiatrists, can do to participate in the policy change to improve our ability to provide appropriate care to our patients in the current climate.

With the increase in life span, there has been a rise in the number of older adults. In 2010, approximately 40 million persons aged 65 years and older lived in the United States and this statistic is projected to increase to 88.5 million by 2050, accounting for 20.2% of the population. The number of older adults with psychiatric disorders is also expected to increase to 15 million by 2030. A significant proportion of these older adults will also have co-morbid medical issues, which may pose challenges. In this session, we will provide an overview of how nursing home facilities and inpatient geriatric psychiatry units are adapting to changes and challenges to provide care for this vulnerable population. In the last 20 years, the number of elderly patients cared for in nursing homes has increased, with facilities expanding to include more beds to respond to this need. The average resident age has also increased, with a corresponding increase in functional limitations, disabilities, cognitive and mental disorders requiring advanced care. Additionally, policy and industry changes over the years have affected the way we provide care for these residents. There has been rapid growth in assisted living facilities in order to provide an alternative care setting for these individuals who require more assistance with activities of daily living. This has led to the development of specialized units that will meet the needs of these residents. Since the implementation of the minimum data set (MDS) system, many prevalence reports have relied on either recorded diagnoses or scales derived from MDS to identify residents with psychiatric conditions. These have also been used as screening tools to identify residents in need of treatment. Available data indicates that there are an inadequate number of geriatric psychiatrists in the USA to care for these older adults with mental illness. The unfortunate consequence has been providers without specialist training or oversight attempting to provide care for residents. This often includes prescribing multiple psychotropic medications that are inappropriate for their symptoms and their age, which can ultimately lead to substantial psychological and functional morbidity. Additionally, these medication side effects and undertreated psychiatric disorders have a significantly negative impact on the residents’ quality of life, disability, and cost of care. Approximately 75-80% of residents in long- term care have a neurocognitive disorder. Although most major neurocognitive disorders are incurable, appropriate comprehensive treatment can substantially improve the quality of life of a resident, their family, and their caregivers. Routine provision of evidence-based care by a mental health care team with expertise in long-term care is essential for maintaining dignity and reducing suffering for these patients. This includes ongoing staff education, discontinuation of inappropriate medications, optimal use of appropriate psychotropic medications and other behavioral interventions. We will review how the geriatric psychiatrist, along with other allied health care professionals, can work as a team in today's highly regulated environment of long term care to improve patient outcomes. With the projection that the older adult population will swell over the next several decades, inpatient geriatric psychiatry facilities have arisen to address the need to provide acute services for these patients. These units appear to have distinct clinically relevant assessment and treatment advantages. They are more comprehensive in their diagnostic assessment of patients, are more likely to provide cognitive assessments and are more cognizant of potential side effects of psychotropic medications. They are also more likely to provide more age-specific aftercare recommendations, which is important in maintaining success after the hospitalization. However these specialized units are limited, expensive, and may encounter a unique set of challenges as the landscape changes for healthcare delivery, reimbursement and managed care. In this session, we will describe key characteristics of a typical geriatric psychiatry unit, discuss advantages of such a unit over a general psychiatry unit, discuss different models of geriatric psychiatry facilities, outline standard admission screening criteria for appropriate patient selection, address treatment challenges associated with the need for rapid improvement in patient psychopathologies and discuss the increased pressures in providing high quality care in the context of reduced length of stays with scarce resources for aftercare. At the end of our presentation, we will briefly showcase the unique characteristics of our inpatient geriatric psychiatry unit.

Background: Sickle cell disease (SCD) has a high mortality rate with unexpected sudden death. So, the purpose of the study was to analyze clinical and/or autopsy findings at the time of death among sickle cell disease (SCD) patients. Methods: This is a retrospective study of SCD patients who died between January 2011 to December 2011 and the morphological evidence of the cause of death was studied in a tertiary care hospital of south Gujarat. The clinical data, including the age, gender, symptoms and the major autopsy findings and cause of death were obtained for each patient that included histopathological examination. Results: A total of 472 autopsies were performed, out of which sickled erythrocytes were detected in 40 cases. The mean age at death was 30 years and a male/female ratio of 3:1 and peak mortality was in the 2 nd and 3 rd decades of life. The common causes of death in this study include vaso-occlusive crisis (45%), infection (40%). The terminal infection was heralded by upper respiratory tract (43.7%) and by gastroenteritis (31.2%). Other causes of death included intracranial haemorrhage (2.5%) and cirrhosis (2.5%). Among the cases of SCD, 10% of deaths were non-haemoglobinopathy related such as fall (accidental), organophosphorus poisoning and snake bite. Conclusion: This study is to precise analysis of causes of death is needed to focus and improve morbidity and mortality in sickle cell disease especially in highly prevalent area and will impact on the overall survival of these patients.

Summary Fidaxomicin is sporicidal and may be associated with a reduced time to resolution of diarrhoea when used to treat patients with Clostridium difficile infection (CDI). This study investigated whether fidaxomicin for treatment of all patients with CDI reduced C. difficile environmental contamination. Surfaces in the rooms of 66 hospitalized patients treated with metronidazole and/or vancomycin and 68 hospitalized patients treated with fidaxomicin were sampled. Patients treated with fidaxomicin were less likely to contaminate their environment (25/68, 36.8%) than patients treated with metronidazole and/or vancomycin (38/66 57.6%) ( P = 0.02). Treatment with fidaxomicin was associated with reduced environmental contamination with C. difficile.

Background: The objective of the study was to study the spectrum of pathological lesions in patients with lung mass and to study correlation between clinical findings, histopathological pattern and immunohistochemical stains in various biopsy specimen for differentiation and typing of tumors. Methods: This retrospective study was done for the period of three years at Department of Pathology, New Civil Hospital, Surat, India, which is a tertiary health care Centre. Here we studied 78 cases of lung biopsy received in formalin, which were subjected to histopathological examination. Immunohistochemistry was performed as and when required. Results: Total 78 lung biopsy specimens were examined. Out of which, 59 cases (75.6%) were neoplastic, 12 cases(15.4%) were non-neoplastic and 7 cases (9%) were inconclusive. The commonest histological type of malignancy was adenocarcinoma which is associated with peripheral mass lesion, female gender and in non-smokers. Commonest non-neoplastic lesion was tuberculosis. Malignancy was seen quite common in patients presented with lung masses in our institute. Conclusions: Lung tumours are quite common in patients presented with mass lesion. Similar to global trend, adenocarcinoma is the commonest histological type now and associated with change in incidence among women, in non-smokers, molecular alteration and prognosis which need further investigation. Immunohistochemistry is helpful in cases which are not accurately subtyped by histomorphology alone.

Background: : Immunohistochemistry play a very important role in modern surgical pathology especially for identification of tumors lacking the evidence of lineage differentiation on the basis of routine Hematoxylin and Eosin (H&E) stain alone. More than 90% of the tumor with diagnostic difficulties by routine H & E stain could be very well classified by using IHC. The aim of present study is to classify and identify MSRCT. Methods: The study was carried out in Department of Pathology, Government Medical College in a period from August 2008 to November 2011. Total 38 cases of MSRCT are selected for Immunohistochemical staining (IHC) and they are classified and categorized accordingly after IHC. Results: Out of 38 cases of MSRCT, there are 18 cases (47.36%) of non-Hodgkin lymphoma, 3 (7.89%) cases of Neuroblastoma, 3 (7.89%) cases of Synovial Sarcoma, 2 (5.26%) cases of Ewing Sarcoma, 2 (5.26%) cases of Undifferentiated neuroendocrine carcinoma, 2 (5.26%) case of Desmoplastic Small Round Cell Tumor, 1 (2.63%) case of Primitive Neuroectodermal Tumor, 1 (2.63%) case of Amelanotic Melanoma (Small cell variant), 1 (2.63%) case of Anaplastic Dysgerminoma, 1 (2.63%) case of Osteosarcoma, 1 (2.63 %) case of Wilm's tumor, 1 (2.63%) case of Dendritic cell Tumor Testis, 1 (2.63%) case of Undifferentiated Nasopharyngeal Carcinoma, 1 (2.63%) case of Embryonal rhabdomyosarcoma. Conclusions: IHC is very valuable tool for adequate and accurate categorization of MSRCT.

Introduction: In the developed world,Clostridium difficile infection (CDI) is the mostimportant cause of nosocomial infectiousdiarrhea. In addition to providingepidemiological data and helping to indicatethat a local outbreak may be occurring,laboratory tests are used to augment clinicaldecisions on individual patients. Very rarely dodiagnostic tests provide results at the point of decision making; in the intervening periodbetween requesting investigations on a patientwith suspected CDI and return of the laboratoryresult, decisions must be made regardingpatient isolation and treatment.Methods: A 22-month, real-world feasibilitystudy was conducted in patients with clinicallysignificant diarrhea, in a London Hospitalbetween March 2011 and January 2013, inthree older persons’ wards and two intensivecare units (ICUs) to determine acceptability,ease of use, change in turnaround time andclinical utility of a rapid, polymerase chainreaction (PCR)-based point-of-care test (POCT)(Cepheid GeneXpert, Sunnyvale, California,USA) for diagnosis of Clostridium difficile. Nursesin the older persons’ ward and laboratorytechnicians in the ICU were trained toperform the test. Residual samples were sent tothe centralized laboratory for parallel testingusing a two-step algorithm.Results: A total of 335 samples were tested usingthe POCT with a median turnaround time of1.85 h compared with 18 h for the centralizedlaboratory test. Overall agreement withcentralized laboratory testing was 98.1%.Discrepant samples were more frequent on elderly wards than ICU. Overall 20/335 (6%)processing errors were encountered and werehighest in the first few months of the study.Significantly more processing errors occurred onthe older persons’ wards 13/102 (12.7%) than onICU 7/271 (2.6%). Older persons’ patients whohad POCT were significantly less likely to have atest requested for bacterial stool culture (3.1% vs.10.9% p = 0.044). This difference was notobserved in the ICU patients. No otherdifferences in ancillary test requesting,mortality or length of stay were observed.Conclusions: The majority of users reportedthat the POCT was easy to perform and was anacceptable part of their job. POCT using thissystem is feasible and acceptable to nursing staffand technicians working within these twohospital-based settings.

267 Background: Over the past two decades the rise in number of oral oncolytics has significantly changed the landscape of cancer treatment. Although these medications offer patients the convenience of home administration, they pose different challenges. Two examples are adherence and education of associated toxicities. In an ongoing effort to provide quality cancer care, the Central Jersey Division of Regional Cancer Care Associates (CJD RCCA) has reconstructed its oral oncolytic program. Methods: In 2016 the CJD of RCCA developed an oral oncolytic program to educate patients on how to safely manage their medications. After the provider sent the prescription, a flowsheet is entered into the EMR, triggering the chemotherapy teaching session and appropriate follow up visits. During this visit patients are provided drug specific educational sheets which include lab monitoring, common adverse effects, and calendars to document adherence. Due to failure of entering flowsheets into the EMR, patients lost to scheduling while waiting for medication delivery, or patient refusal of teaching, compliance remained low. The program was modified to include running a daily report to identify all new oral oncolytic prescriptions. Schedulers are immediately calling patients to schedule a teaching session. A field in the EMR was created to document “patient refusal”. By making these improvements, CJD RCCA has increased compliance along with upholding quality care for patients receiving oral oncolytics. Results: Since re-structuring the program the RCCA CJD APNs have performed teaching on 124 of 172 (72%) patients from January to May of 2019. This is compared to only 79 of 144 (54%) in all of 2018. Of the patients identified in 2019 30 refused, 13 were completed by the physician, and 5 expired. This dramatic rise of 18% is largely due to running the daily oral oncolytic reports and entering the flowsheet into the EMR. Conclusions: Patient compliance has increased by identifying new patients and prompt enrollment into the program. It provides them the tools to safely navigate through treatment. At CJD RCCA, the oral oncolytic program is vital to ensuring positive clinical outcomes for these patients.

Clostridium difficile was recovered from 33 (34%) of 98 rooms of patients who were excretors compared with 36 (49%) of 73 rooms of patients with active infection. Not all laboratory algorithms can distinguish between these 2 groups, yet both may be a significant source for ongoing transmission.Infect Control Hosp Epidemiol 2015;36(8): 975–977

Herpes simplex virus 1 (HSV-1) and HSV-2, two closely related neurotropic human herpesviruses, achieve neurotropism through ICP34.5, a major viral neurovirulence factor. In this report, in addition to the full-length 38-kDa protein (ICP34.5α), we identified a 28-kDa novel form of ICP34.5 (ICP34.5β) in HSV-2-infected cells. ICP34.5β is translated from unspliced ICP34.5 mRNA, with the retained intron introducing a premature stop codon. Thus, ICP34.5β lacks the C-terminal conserved GADD34 domain but includes 19 additional amino acids encoded by the intron. Although a fraction of both HSV-2 ICP34.5 proteins are detected in the nucleolus, ICP34.5α is predominantly located in cytoplasm, and ICP34.5β is mainly detected more diffusely in the nucleus. ICP34.5β is unable to counteract PKR-mediated eIF2 phosphorylation but does not interfere with ICP34.5α's function in this process. Efficient expression of ICP34.5β in cell culture assays is dependent on viral infection or expression of ICP27, a multifunctional immediate-early gene. The effect of ICP27 on the ICP34.5β protein level is attributed to its selective inhibition of ICP34.5 splicing, which results in increased expression of ICP34.5β but a reduced level of ICP34.5α. The C- terminal KH3 domain but not the RNA binding domain of ICP27 is required for its specific inhibition of ICP34.5 splicing and promotion of ICP34.5β expression. Our results suggest that the expression of ICP34.5α and ICP34.5β is tightly regulated in HSV-2 and likely contributes to viral pathogenesis.