Clinical and Epidemiological Correlates of Genotypes within the Mycobacterium avium Complex Defined by Restriction and Sequence Analysis of hsp65

Species identification of isolates of the Mycobacterium avium complex (MAC) remains a difficult task. Although M. avium and Mycobacterium intracellulare can be identified with expensive, commercially available probes, many MAC isolates remain unresolved, including those representing Mycobacterium lentiflavum as well as other potentially undefined species. PCR restriction analysis (PRA) of the hsp65 gene has been proposed as a rapid and inexpensive approach. We applied PRA to 278 MAC isolates, including 126 from blood of human immunodeficiency virus (HIV)-infected patients, 59 from sputum of HIV-negative patients with chronic obstructive pulmonary disease, 88 from environmental sources, and 5 pulmonary isolates from a different study. A total of 15 different PRA patterns were observed. For 27 representative isolates, a 441-bp fragment of the hsp65 gene was sequenced; based on 54 polymorphic sites, 18 different alleles were defined, including 12 alleles not previously reported. Species and phylogenetic relationships were more accurately defined by sequencing than by PRA or commercial probe. The distribution of PRA types and, by implication, phylogenetic lineages among blood isolates was significantly different from that for pulmonary and environmental isolates, suggesting that particular lineages have appreciably greater virulence and invasive potential

Virulence and antimicrobial resistance determinants of verotoxigenic Escherichia coli (VTEC) and of ESBL-producing multidrug resistant E. coli from foods of animal origin illegally imported to Europe

Microbial risk due to illegal food import has not been investigated so far. Here we aimed to reveal frequency, phenotype and genotype of verotoxigenic E. coli (VTEC) and ESBL-producing multidrug resistant (MDR) E. coli isolated from foods of animal origin confiscated at the EU airport borders. Of the 1500 food samples confiscated at the airports of Austria, Germany and Slovenia, the most frequent were cheese and meat products primarily from Turkey and from Balkan countries. The VTEC bacteria were isolated using ISO 16654:2001 for O157 and Ridascreen® ELISA based PCR testing of stx genes or ISO/ TS13136 for non-O157 VTEC, resulting in 15 isolates of VTEC (1%). In addition 600 samples from the Vienna airport were also tested for ESBL-producing MDR E. coli, using cefotaxime-McConkey agar. We identified 14 E. coli strains as ESBL/MDR E. coli. (0,9%) for phenotyping for antimicrobial resistance and for genotypiing by microarray (Identibac®,AMR05). The 15 VTEC isolates were phenotyped as Stx toxin producing non-O157 strain. Only one isolate, from Turkish cheese, proved to be EHEC (O26:H46). The remaining 14 strains represent uncommon VTEC serotypes with stx1 and/or stx2 genes. Microarray analysis (Identibac®, Ec03) revealed a wide range of other non-LEE encoding virulence genes. Pulsed field electrophoresis (PFGE) showed high genetic diversity of the strains. Multilocus sequence typing (MLST) established three new ST types (ST4505, 4506 and 4507) in the MLST database, and indicated the existence of 5 small clusters with no relation to origin or serotype/genotype of the strains, but representing several human-related ST types. All VTEC isolates were sensitive to 18 antimicrobials relevant to human and/or animal health, and did not contain resistance genes. ESB/MDR E. coli were resistant to at least 3 classes of antimicrobials. Microarray analysis detected TEM-1 in all but one strain and a variety of genes encoding resistances to other ESBLs (CTXM-1, OXA-1), trimethromprim, tetracycline, aminoglycosides and class1/class2 integrons (8/14 isolates). E.coli virulence microarray detected 2-6 virulence genes in all but one MDR E. coli, and one of the strains qualified as an atypical EPEC . Even though the frequency and attributes of isolated VTEC and ESBL/MDR E. coli did not represent an immediate major risk through illegal food import for the countries involved, it is suggested that the unusual serovars of VTEC as well as the virulence and antimicrobial resistance determinants of ESBL/MDR E. coli detected here, may indicate a future emerging threat by strains in illegally imported foods. Acknowledgement is due to: EU FP7 PROMISE (Grant No: 265877), to Dr. Mária Herpay, National Institute for Epidemiology, Budapest

Fast-timing measurements in the ground-state band of Pd114

Using a hybrid Gammasphere array coupled to 25 LaBr3(Ce) detectors, the lifetimes of the first three levels of the yrast band in Pd-114, populated via Cf-252 decay, have been measured. The measured lifetimes are tau(2+) = 103(10) ps, tau(4+) = 22(13) ps, and tau(6+) <= 10 ps for the 2(1)(+), 4(1)(+), and 6(1)(+) levels, respectively. Palladium-114 was predicted to be the most deformed isotope of its isotopic chain, and spectroscopic studies have suggested it might also be a candidate nucleus for low-spin stable triaxiality. From the lifetimes measured in this work, reduced transition probabilities B(E2; J -> J - 2) are calculated and compared with interacting boson model, projected shell model, and collective model calculations from the literature. The experimental ratio R-B(E2) = B(E2; 4(1)(+) -> 2(1)(+))/B(E2; 2(1)(+) -> 0(1)(+)) = 0.80(42) is measured for the first time in Pd-114 and compared with the known values R-B(E2) in the palladium isotopic chain: the systematics suggest that, for N = 68, a transition from gamma-unstable to a more rigid gamma-deformed nuclear shape occurs

Comparative Transmissibility of SARS-CoV-2 Variants Delta and Alpha in New England, USA

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Delta variant quickly rose to dominance in mid-2021, displacing other variants, including Alpha. Studies using data from the United Kingdom and India estimated that Delta was 40-80% more transmissible than Alpha, allowing Delta to become the globally dominant variant. However, it was unclear if the ostensible difference in relative transmissibility was due mostly to innate properties of Delta\u27s infectiousness or differences in the study populations. To investigate, we formed a partnership with SARS-CoV-2 genomic surveillance programs from all six New England US states. By comparing logistic growth rates, we found that Delta emerged 37-163% faster than Alpha in early 2021 (37% Massachusetts, 75% New Hampshire, 95% Maine, 98% Rhode Island, 151% Connecticut, and 163% Vermont). We next computed variant-specific effective reproductive numbers and estimated that Delta was 58-120% more transmissible than Alpha across New England (58% New Hampshire, 68% Massachusetts, 76% Connecticut, 85% Rhode Island, 98% Maine, and 120% Vermont). Finally, using RT-PCR data, we estimated that Delta infections generate on average ∼6 times more viral RNA copies per mL than Alpha infections. Overall, our evidence indicates that Delta\u27s enhanced transmissibility could be attributed to its innate ability to increase infectiousness, but its epidemiological dynamics may vary depending on the underlying immunity and behavior of distinct populations

Comparative transmissibility of SARS-CoV-2 variants Delta and Alpha in New England, USA.

The SARS-CoV-2 Delta variant rose to dominance in mid-2021, likely propelled by an estimated 40%-80% increased transmissibility over Alpha. To investigate if this ostensible difference in transmissibility is uniform across populations, we partner with public health programs from all six states in New England in the United States. We compare logistic growth rates during each variant\u27s respective emergence period, finding that Delta emerged 1.37-2.63 times faster than Alpha (range across states). We compute variant-specific effective reproductive numbers, estimating that Delta is 63%-167% more transmissible than Alpha (range across states). Finally, we estimate that Delta infections generate on average 6.2 (95% CI 3.1-10.9) times more viral RNA copies per milliliter than Alpha infections during their respective emergence. Overall, our evidence suggests that Delta\u27s enhanced transmissibility can be attributed to its innate ability to increase infectiousness, but its epidemiological dynamics may vary depending on underlying population attributes and sequencing data availability

Development of an amplicon-based sequencing approach in response to the global emergence of mpox

The 2022 multicountry mpox outbreak concurrent with the ongoing Coronavirus Disease 2019 (COVID-19) pandemic further highlighted the need for genomic surveillance and rapid pathogen whole-genome sequencing. While metagenomic sequencing approaches have been used to sequence many of the early mpox infections, these methods are resource intensive and require samples with high viral DNA concentrations. Given the atypical clinical presentation of cases associated with the outbreak and uncertainty regarding viral load across both the course of infection and anatomical body sites, there was an urgent need for a more sensitive and broadly applicable sequencing approach. Highly multiplexed amplicon-based sequencing (PrimalSeq) was initially developed for sequencing of Zika virus, and later adapted as the main sequencing approach for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Here, we used PrimalScheme to develop a primer scheme for human monkeypox virus that can be used with many sequencing and bioinformatics pipelines implemented in public health laboratories during the COVID-19 pandemic. We sequenced clinical specimens that tested presumptively positive for human monkeypox virus with amplicon-based and metagenomic sequencing approaches. We found notably higher genome coverage across the virus genome, with minimal amplicon drop-outs, in using the amplicon-based sequencing approach, particularly in higher PCR cycle threshold (Ct) (lower DNA titer) samples. Further testing demonstrated that Ct value correlated with the number of sequencing reads and influenced the percent genome coverage. To maximize genome coverage when resources are limited, we recommend selecting samples with a PCR Ct below 31 Ct and generating 1 million sequencing reads per sample. To support national and international public health genomic surveillance efforts, we sent out primer pool aliquots to 10 laboratories across the United States, United Kingdom, Brazil, and Portugal. These public health laboratories successfully implemented the human monkeypox virus primer scheme in various amplicon sequencing workflows and with different sample types across a range of Ct values. Thus, we show that amplicon-based sequencing can provide a rapidly deployable, cost-effective, and flexible approach to pathogen whole-genome sequencing in response to newly emerging pathogens. Importantly, through the implementation of our primer scheme into existing SARS-CoV-2 workflows and across a range of sample types and sequencing platforms, we further demonstrate the potential of this approach for rapid outbreak response.This publication was made possible by CTSA Grant Number UL1 TR001863 from the National Center for Advancing Translational Science (NCATS), a component of the National Institutes of Health (NIH) awarded to CBFV. INSA was partially funded by the HERA project (Grant/ 2021/PHF/23776) supported by the European Commission through the European Centre for Disease Control (to VB).info:eu-repo/semantics/publishedVersio

Cumulative Prognostic Score Predicting Mortality in Patients Older Than 80 Years Admitted to the ICU.

OBJECTIVES: To develop a scoring system model that predicts mortality within 30 days of admission of patients older than 80 years admitted to intensive care units (ICUs). DESIGN: Prospective cohort study. SETTING: A total of 306 ICUs from 24 European countries. PARTICIPANTS: Older adults admitted to European ICUs (N = 3730; median age = 84 years [interquartile range = 81-87 y]; 51.8% male). MEASUREMENTS: Overall, 24 variables available during ICU admission were included as potential predictive variables. Multivariable logistic regression was used to identify independent predictors of 30-day mortality. Model sensitivity, specificity, and accuracy were evaluated with receiver operating characteristic curves. RESULTS: The 30-day-mortality was 1562 (41.9%). In multivariable analysis, these variables were selected as independent predictors of mortality: age, sex, ICU admission diagnosis, Clinical Frailty Scale, Sequential Organ Failure Score, invasive mechanical ventilation, and renal replacement therapy. The discrimination, accuracy, and calibration of the model were good: the area under the curve for a score of 10 or higher was .80, and the Brier score was .18. At a cut point of 10 or higher (75% of all patients), the model predicts 30-day mortality in 91.1% of all patients who die. CONCLUSION: A predictive model of cumulative events predicts 30-day mortality in patients older than 80 years admitted to ICUs. Future studies should include other potential predictor variables including functional status, presence of advance care plans, and assessment of each patient's decision-making capacity

Relationship between the Clinical Frailty Scale and short-term mortality in patients ≥ 80 years old acutely admitted to the ICU: a prospective cohort study.

BACKGROUND: The Clinical Frailty Scale (CFS) is frequently used to measure frailty in critically ill adults. There is wide variation in the approach to analysing the relationship between the CFS score and mortality after admission to the ICU. This study aimed to evaluate the influence of modelling approach on the association between the CFS score and short-term mortality and quantify the prognostic value of frailty in this context. METHODS: We analysed data from two multicentre prospective cohort studies which enrolled intensive care unit patients ≥ 80 years old in 26 countries. The primary outcome was mortality within 30-days from admission to the ICU. Logistic regression models for both ICU and 30-day mortality included the CFS score as either a categorical, continuous or dichotomous variable and were adjusted for patient's age, sex, reason for admission to the ICU, and admission Sequential Organ Failure Assessment score. RESULTS: The median age in the sample of 7487 consecutive patients was 84 years (IQR 81-87). The highest fraction of new prognostic information from frailty in the context of 30-day mortality was observed when the CFS score was treated as either a categorical variable using all original levels of frailty or a nonlinear continuous variable and was equal to 9% using these modelling approaches (p < 0.001). The relationship between the CFS score and mortality was nonlinear (p < 0.01). CONCLUSION: Knowledge about a patient's frailty status adds a substantial amount of new prognostic information at the moment of admission to the ICU. Arbitrary simplification of the CFS score into fewer groups than originally intended leads to a loss of information and should be avoided. Trial registration NCT03134807 (VIP1), NCT03370692 (VIP2)