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BackgroundDue to their prevalence worldwide, the β-lactamases CTX-M and plasmid-mediated CMY-2 are important antimicrobial resistance enzymes in a clinical setting. While culture- and PCR-based detection methods exist for these targets, they are time consuming and require specialist equipment and trained personnel to carry out.MethodsIn this study, three rapid diagnostic single-plex and a prototype triplex assay were developed, using recombinase polymerase amplification with lateral flow detection (RPA-LF), and tested for their sensitivity and specificity using two isolate DNA panels (n = 90 and n = 120 isolates). In addition, the RPA-LF assays were also tested with a small number of faecal extract samples (n = 18).ResultsThe RPA-LF assays were able to detect blaCXT-M-group-1, blaCTX-M-group-9 and blaCMY-2-type variants with high sensitivity (82.1%–100%) and specificity (100%) within a short turnaround time (15–20 min for amplification and detection).ConclusionsRPA-LF assays developed in this study have the potential to be used at or close to the point of care, as well as in low-resource settings, producing rapid results to support healthcare professionals in their treatment decisions.

Background A novel human parechovirus 3 Australian recombinant (HPeV3-AR) strain emerged in 2013 and coincided with biennial outbreaks of sepsis-like illnesses in infants. We evaluated the molecular evolution of the HPeV3-AR strain and its association with severe HPeV infections. Methods HPeV3-positive samples collected from hospitalized infants aged 5–252 days in 2 Australian states (2013–2020) and from a community-based birth cohort (2010–2014) were sequenced. Coding regions were used to conduct phylogenetic and evolutionary analyses. A recombinant-specific polymerase chain reaction was designed and utilized to screen all clinical and community HPeV3-positive samples. Results Complete coding regions of 54 cases were obtained, which showed the HPeV3-AR strain progressively evolving, particularly in the 3′ end of the nonstructural genes. The HPeV3-AR strain was not detected in the community birth cohort until the initial outbreak in late 2013. High-throughput screening showed that most (>75%) hospitalized HPeV3 cases involved the AR strain in the first 3 clinical outbreaks, with declining prevalence in the 2019–2020 season. The AR strain was not statistically associated with increased clinical severity among hospitalized infants. Conclusions HPeV3-AR was the dominant strain during the study period. Increased hospital admissions may have been from a temporary fitness advantage and/or increased virulence.

Isotemporal substitution model is a powerful tool to explore the real association between physical behavior and health outcomes, which has the potential of the application in large-scale cohort study. This paper systematically introduces the principle of isotemporal substitution model and its implementation method in specific analysis to provide analytical ideas for the epidemiological research related to physical behavior in China. The baseline data of Regional Ethic Cohort Study in Northwest China conducted in Shaanxi province were used to analyze the relationship between physical behavior and cardiovascular disease with single-factor model, partition model and isotemporal substitution model. The advantages and disadvantages of different models were compared, and the advantages of isotemporal substitution model in quantifying physical activity health risk were introduced. Isotemporal substitution model could qualify physical behavior and health outcomes, which has wide application value in epidemiological research.等时替代模型（ISM）是探索身体行为与健康结局之间真实关联的有力工具，在大型自然人群队列研究中具有突出的应用潜力。本文系统介绍ISM的原理和具体分析中的实现方法，以期为国内身体行为相关的流行病学研究提供分析思路。通过西北区域自然人群队列研究陕西省人群基线调查数据，分别采用单一活动模型、分割模型以及ISM分析身体活动与心血管疾病的关联，比较不同模型优缺点，展示ISM在量化身体活动健康风险研究中的优势。ISM可以较好量化身体活动与健康结局的真实关联，在流行病学研究中具有广泛的应用价值。.

BackgroundAlthough aging correlates with a worse prognosis for Covid-19, super elderly still unvaccinated individuals presenting mild or no symptoms have been reported worldwide. Most of the reported genetic variants responsible for increased disease susceptibility are associated with immune response, involving type I IFN immunity and modulation; HLA cluster genes; inflammasome activation; genes of interleukins; and chemokines receptors. On the other hand, little is known about the resistance mechanisms against SARS-CoV-2 infection. Here, we addressed polymorphisms in the MHC region associated with Covid-19 outcome in super elderly resilient patients as compared to younger patients with a severe outcome.MethodsSARS-CoV-2 infection was confirmed by RT-PCR test. Aiming to identify candidate genes associated with host resistance, we investigated 87 individuals older than 90 years who recovered from Covid-19 with mild symptoms or who remained asymptomatic following positive test for SARS-CoV-2 as compared to 55 individuals younger than 60 years who had a severe disease or died due to Covid-19, as well as to the general elderly population from the same city. Whole-exome sequencing and an in-depth analysis of the MHC region was performed. All samples were collected in early 2020 and before the local vaccination programs started.ResultsWe found that the resilient super elderly group displayed a higher frequency of some missense variants in the MUC22 gene (a member of the mucins’ family) as one of the strongest signals in the MHC region as compared to the severe Covid-19 group and the general elderly control population. For example, the missense variant rs62399430 at MUC22 is two times more frequent among the resilient super elderly (p = 0.00002, OR = 2.24).ConclusionSince the pro-inflammatory basal state in the elderly may enhance the susceptibility to severe Covid-19, we hypothesized that MUC22 might play an important protective role against severe Covid-19, by reducing overactive immune responses in the senior population.

Carbapenemase-producing organisms (CPOs) pose a serious clinical threat and rapid detection tools are essential to aid in patient management. We developed rapid and simple molecular tests to detect blaNDM-type and blaVIM-type carbapenemase genes using recombinase polymerase amplification (RPA) combined with a lateral flow detection. The tests could provide results in approximately 15 min when using DNA extracts, with limits of detection of 9.2 copies/μl for the blaNDM-type assay and 7.5 copies/μl for blaVIM-type assay, and successfully detected all isolates harbouring the carbapenemase encoding genes in a panel of 57 isolates. These RPA tests may be suitable for use in low-resource settings to tailor rapid implementation of infection control precautions and antibiotic stewardship.

Background Volunteer infection studies have become a standard model for evaluating drug efficacy against Plasmodium infections. Molecular techniques such as qPCR are used in these studies due to their ability to provide robust and accurate estimates of parasitaemia at increased sensitivity compared to microscopy. The validity and reliability of assays need to be ensured when used to evaluate the efficacy of candidate drugs in clinical trials. Methods A previously described 18S rRNA gene qPCR assay for quantifying Plasmodium falciparum in blood samples was evaluated. Assay performance characteristics including analytical sensitivity, reportable range, precision, accuracy and specificity were assessed using experimental data and data compiled from phase 1 volunteer infection studies conducted between 2013 and 2019. Guidelines for validation of laboratory-developed molecular assays were followed. Results The reportable range was 1.50 to 6.50 log10 parasites/mL with a limit of detection of 2.045 log10 parasites/mL of whole blood based on a parasite diluted standard series over this range. The assay was highly reproducible with minimal intra-assay (SD = 0.456 quantification cycle (Cq) units [0.137 log10 parasites/mL] over 21 replicates) and inter-assay (SD = 0.604 Cq units [0.182 log10 parasites/mL] over 786 qPCR runs) variability. Through an external quality assurance program, the QIMR assay was shown to generate accurate results (quantitative bias + 0.019 log10 parasites/mL against nominal values). Specificity was 100% after assessing 164 parasite-free human blood samples. Conclusions The 18S rRNA gene qPCR assay is specific and highly reproducible and can provide reliable and accurate parasite quantification. The assay is considered fit for use in evaluating drug efficacy in malaria clinical trials.

Cervical cancer is the most common gynecological malignant carcinoma in Chinese women, which seriously threatens women's health. Lymph node metastasis is the most important factor affecting the prognosis and adjuvant treatment of patients with early cervical cancer. Conventional systematic lymphadenectomy has always been a reliable method to detect lymph node metastasis. However, this procedure may affect the quality of life of patients because of high risks of complications. Recently, sentinel lymph node biopsy (SLNB) has been researched aiming to assess the status of pelvic lymph node metastasis intraoperatively, avoid unnecessary lymphadenectomy and achieve the goal of individualized diagnosis and treatment. Exploring efficient tracing methods and pathological ultra-staging is the key to the clinical application of SLNB for cervical cancer, and understanding the latest relevant clinical research progress will help SLNB to be applied in the clinic as soon as possible to benefit patients with cervical cancer.子宫颈癌是中国女性最常见的妇科恶性肿瘤，严重危害女性健康。淋巴结转移是影响早期子宫颈癌预后和辅助治疗的最主要因素。传统的盆腔淋巴结清扫术一直是确定淋巴结转移情况的可靠手段，但其并发症发生率高，严重影响患者的生活质量。前哨淋巴结活检术(SLNB)的应用旨在高效、准确评估盆腔淋巴结转移状态，以避免不必要的淋巴结清扫术，达到个体化诊疗目的。探索高效的前哨淋巴结示踪方法和病理学检测方法是子宫颈癌SLNB临床应用的关键，了解最新相关临床研究进展，有助于SLNB早日应用于临床，使子宫颈癌患者受益。.

Ciprofol is a newly developed intravenous sedative-hypnotic drug. The objective of the study was to prove whether ciprofol was non-inferior to propofol for the successful induction of general anesthesia. The ideal post-induction sedation level was assessed by comparing patients' clinical symptoms and their hemodynamic effects in responding to noxious stimuli, mostly tracheal intubation and bispectral index (BIS) alterations following ciprofol/propofol administration.In this multi-center, randomized, double-blind phase 3 trial, selective surgery patients were randomly assigned in a 1:1 ratio to either ciprofol 0.4 mg/kg (n = 88) or propofol 2.0 mg/kg (n = 88) groups. The primary endpoint was the percentage of patients with successful anesthesia inductions. Secondary endpoints included the times to successful induction of general anesthesia and loss of the eyelash reflex, changes in BIS, as well as safety indicators.The anesthesia induction success rates for both ciprofol 0.4 mg/kg and propofol 2 mg/kg groups were 100.0%, with a 95% CI lower success limit of -4.18% difference between the two groups, indicating that ciprofol was non-inferior to propofol. For secondary outcomes, the average time to successful anesthesia and loss of the eyelash reflex were 0.91 min and 0.80 min for ciprofol and 0.80 min and 0.71 min for propofol, respectively. The pattern of BIS changes with ciprofol was similar to propofol and stable during the anesthesia maintenance period. Safety was comparable with 88.6% TEAEs in the ciprofol group compared to 95.5% in the propofol group. The incidence of injection pain was significantly lower in the ciprofol group compared to the propofol group (6.8% vs. 20.5%, p0.05). In addition, the patients treated with ciprofol had a lesser increase in blood pressure and heart rate, and fewer cases with BIS60 within 15 min of intravenous administration, which indicated that ciprofol may provide a better ideal sedation level during the post-induction period under an equivalent dosing regimen to propofol.Ciprofol for patients undergoing selective surgery is a new option for the induction of general anesthesia.

The rapid development of urbanisation and the ever-changing consumers’ demands are constantly changing the urban logistics industry, imposing challenges on logistics service providers to improve cu...

Summary Background Perhaps, as never before, we need innovators. With our growing population numbers, and with increasing pressures on our education systems, are we in danger of becoming more rigid and formulaic and increasingly inhibiting innovation? When young can we predict who will become the great innovators? For example, in medicine, who will change clinical practice? Aims We therefore determined to assess whether the current academic excellence approach to medical school entrance would have captured previous great innovators in medicine, assuming that they should all have well fulfilled current entrance requirements. Methods The authors assembled a list of 100 great medical innovators which was then approved, rejected or added to by a jury of 12 MD fellows of the Royal Society of Canada. Two reviewers, who had taken both the past and present Medical College Admission Test as part of North American medical school entrance requirements, independently assessed each innovator’s early life educational history in order to predict the innovator’s likely success at medical school entry, assuming excellence in all entrance requirements. Results Thirty-one percent of the great medical innovators possessed no medical degree and 24% would likely be denied entry to medical school by today’s standards (e.g. had a history of poor performance, failure, dropout or expulsion) with only 24% being guaranteed entry. Even if excellence in only one topic was required, the figure would only rise to 41% certain of medical school entry. Conclusion These data show that today’s medical school entry standards would have barred many great innovators and raise questions about whether we are losing medical innovators as a consequence. Our findings have important implications for promoting flexibility and innovation for medical education, and for promoting an environment for innovation in general.

BACKGROUNDInterventions that interrupt Plasmodium vivax transmission or eliminate dormant P. vivax liver-stage parasites will be essential for malaria elimination. Development of these interventions has been hindered by the lack of P. vivax in vitro culture and could be accelerated by a safe and reproducible clinical model in malaria-naive individuals.METHODSHealthy, malaria-naive adults were enrolled in 2 studies to assess the safety, infectivity, and transmissibility of a new P. vivax isolate. Participants (Study 1, n = 2; Study 2, n = 24) were inoculated with P. vivax-infected red blood cells to initiate infection, and were treated with artemether-lumefantrine (Study 1) or chloroquine (Study 2). Primary endpoints were safety and infectivity of the new isolate. In Study 2, transmission to mosquitoes was also evaluated using mosquito feeding assays, and sporozoite viability was assessed using in vitro cultured hepatocytes.RESULTSParasitemia and gametocytemia developed in all participants and was cleared by antimalarial treatment. Adverse events were mostly mild or moderate and none were serious. Sixty-nine percent of participants (11/16) were infectious to Anopheles mosquitoes at peak gametocytemia. Mosquito infection rates reached 97% following membrane feeding with gametocyte-enriched blood, and sporozoites developed into liver-stage schizonts in culture.CONCLUSIONWe have demonstrated the safe, reproducible, and efficient transmission of P. vivax gametocytes from humans to mosquitoes, and have established an experimental model that will accelerate the development of interventions targeting multiple stages of the P. vivax life cycle.TRIAL REGISTRATIONACTRN12614000930684 and ACTRN12616000174482.FUNDING(Australian) National Health and Medical Research Council Program Grant 1132975 (Study 1). Bill and Melinda Gates Foundation (OPP1111147) (Study 2).

Made available in DSpace on 2022-04-29T08:46:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Recurrence of COVID-19 in recovered patients has been increasingly reported. However, the immune mechanisms behind the recurrence have not been thoroughly investigated. The presence of neutralizing antibodies (nAbs) in recurrence/reinfection cases suggests that other types of immune response are involved in protection against recurrence. Here, we investigated the innate type I/III interferon (IFN) response, binding and nAb assays and T-cell responses to severe acute respiratory distress syndrome coronavirus 2 (SARS-CoV-2) with IFN gamma (IFNγ) enzyme-linked spot assay (ELISPOT) in three pairs of young adult monozygotic (MZ) twins with previous confirmed COVID-19, one of them presenting a severe recurrence four months after the initial infection. Twin studies have been of paramount importance to comprehend the immunogenetics of infectious diseases. Each MZ twin pair was previously exposed to SARS-CoV-2, as seen by clinical reports. The six individuals presented similar overall recovered immune responses except for the recurrence case, who presented a drastically reduced number of recognized SARS-CoV-2 T-cell epitopes on ELISPOT as compared to her twin sister and the other twin pairs. Our results suggest that the lack of a broad T-cell response to initial infection may have led to recurrence, emphasizing that an effective SARS-CoV-2-specific T-cell immune response is key for complete viral control and avoidance of clinical recurrence of COVID-19. Human Genome and Stem Cell Research Center (HUG-CELL) Biosciences Institute Universidade de São Paulo, SP Laboratory of Immunology Heart Institute (InCor) Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HCFMUSP), SP Institute for Investigation in Immunology Instituto Nacional de Ciência e Tecnologia - Iii-INCT, SP Division of Clinical Immunology and Allergy Department of Medicine Faculdade de Medicina da Universidade de São Paulo (FMUSP), SP Department of Microbiology Immunology and Parasitology Universidade Federal de São Paulo (UNIFESP/EPM), SP Department of Parasitology Biosciences Institute Universidade de São Paulo, SP School of Medicine Universidade Estadual Paulista (UNESP), SP Fleury Laboratory, SP School of Medicine Universidade Estadual Paulista (UNESP), SP

We develop a compact theory that can be applied to a variety of time-varying dispersive materials. The continuous wave reflection and transmission coefficients are replaced with equivalent operator expressions. In addition to comparing this approach to existing numerical and analytical techniques, we find that the eigenfunctions of these operators represent pulses that do not change their spectra after interaction with the time-varying, dispersive material. In addition, the poles of these operators represent the non-time harmonic bound states of the system., Comment: 6 pages, 3 figures

Made available in DSpace on 2022-04-29T08:46:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-09-28 Center for Information Technology Center for Scientific Review National Institutes of Health Office of Extramural Research, National Institutes of Health Office of Research Infrastructure Programs, National Institutes of Health Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Despite the high number of individuals infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) who develop coronavirus disease 2019 (COVID-19) symptoms worldwide, many exposed individuals remain asymptomatic and/or uninfected and seronegative. This could be explained by a combination of environmental (exposure), immunological (previous infection), epigenetic, and genetic factors. Aiming to identify genetic factors involved in immune response in symptomatic COVID-19 as compared to asymptomatic exposed individuals, we analyzed 83 Brazilian couples where one individual was infected and symptomatic while the partner remained asymptomatic and serum-negative for at least 6 months despite sharing the same bedroom during the infection. We refer to these as “discordant couples”. We performed whole-exome sequencing followed by a state-of-the-art method to call genotypes and haplotypes across the highly polymorphic major histocompatibility complex (MHC) region. The discordant partners had comparable ages and genetic ancestry, but women were overrepresented (65%) in the asymptomatic group. In the antigen-presentation pathway, we observed an association between HLA-DRB1 alleles encoding Lys at residue 71 (mostly DRB1*03:01 and DRB1*04:01) and DOB*01:02 with symptomatic infections and HLA-A alleles encoding 144Q/151R with asymptomatic seronegative women. Among the genes related to immune modulation, we detected variants in MICA and MICB associated with symptomatic infections. These variants are related to higher expression of soluble MICA and low expression of MICB. Thus, quantitative differences in these molecules that modulate natural killer (NK) activity could contribute to susceptibility to COVID-19 by downregulating NK cell cytotoxic activity in infected individuals but not in the asymptomatic partners. Department of Pathology School of Medicine São Paulo State University (UNESP) Molecular Genetics and Bioinformatics Laboratory–Experimental Research Unit School of Medicine São Paulo State University (UNESP) Human Genome and Stem Cell Research Center University of São Paulo Department of Genetics and Evolutionary Biology Biosciences Institute University of São Paulo Centro Universitário Sudoeste Paulista Departamento de Química Faculdade de Filosofa Ciências e Letras de Ribeirão Preto Universidade de São Paulo Department of Clinical and Toxicological Analyses School of Pharmaceutical Sciences University of São Paulo Departamento de Clínica Médica Disciplina de Alergia e Imunologia Clínica Faculdade de Medicina da Universidade de São Paulo Laboratório de Imunologia Instituto do Coração (InCor) LIM19 Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HCFMUSP) Instituto de Investigação em Imunologia - Instituto Nacional de Ciências e Tecnologia-iii-INCT Department of Pathology School of Medicine São Paulo State University (UNESP) Molecular Genetics and Bioinformatics Laboratory–Experimental Research Unit School of Medicine São Paulo State University (UNESP) CAPES: 001 FAPESP: 2013/08028-1 FAPESP: 2013/17084-0 FAPESP: 2014/50931-3 FAPESP: 2017/19223-0 FAPESP: 2019/19998-8 FAPESP: 2020/09702-1 CNPq: 465355/2014-5

The equilibrium sediment exchange process is defined as instantaneous deposition of suspended sediment to the streambed countered by equal erosion of sediment from the streambed. Equilibrium exchange has rarely been included in sediment transport studies but is needed when the sediment continuum is used to investigate the earth’s critical zone. Numerical modeling in the watershed uplands and stream corridor simulates sediment yield and sediment source partitioning for the Upper South Elkhorn watershed in Kentucky, USA. We simulate equilibrium exchange when upland-derived sediment simultaneously deposits to the streambed while streambed sediments erode. Sediment fingerprinting with stable carbon isotopes allowed constraint of the process in a gently rolling watershed. Carbon isotopes work well to partition upland sediment versus streambed sediment because sediment deposited in the streambed accrues a unique autotrophic, i.e., algal, fingerprint. Stable nitrogen isotopes do not work well to partition the sources in this study because the nitrogen isotope fingerprint of algae falls in the middle of the nitrogen isotope fingerprint of upland sediment. The source of sediment depends on flow intensity for the gently rolling watershed. Streambed sediments dominate the fluvial load for low and moderate events, while upland sediments become increasingly important during high flows and extreme events. We used sediment fingerprinting results to calibrate the equilibrium sediment exchange rate in the watershed sediment transport model. Our sediment fingerprinting and modeling evidence suggest equilibrium sediment exchange is a substantial process occurring in the system studied. The process does not change the sediment load or streambed sediment storage but does impact the quality of sediment residing in the streambed. Therefore, we suggest equilibrium sediment exchange should be considered when the sediment continuum is used to investigate the critical zone. We conclude the paper by outlining future research priorities for coupling sediment fingerprinting with watershed modeling.

免疫检查点抑制剂是肿瘤免疫治疗的一类药物，在肿瘤治疗中有很好的应用前景。该药可引起多系统不良反应，其中心血管不良反应虽不常见，但是致死率高，需重视。其常见的心血管不良反应包括心肌炎、心力衰竭、心律失常、急性冠状动脉事件等。该文总结了免疫检查点抑制剂所致心血管不良反应的机制、临床表现及管理，以期提高临床医生对其的警惕性和应对能力。.