Test Sequences Research Articles

Assessment of age-related features of the condition of heavy-duty truck drivers based on spectral analysis of EEG-ECG and stress tests during mass occupational examinations

The authors studied the physiological characteristics of drivers of heavy-duty coal mine vehicles based on spectral analysis of EEG-ECG and stress tests, which have prognostic value in professional activity. The study aims to assess the functional condition of heavy-duty truck drivers of different age groups based on a comprehensive spectral analysis of EEG-ECG under occupational examinations. During the survey, the authors have studied the spectral parameters of the EEG‒ECG in 300 male drivers aged 25 to 66 years. Specialists have performed EEG registration when placing electrodes in the 10–20 system. The EEG analysis was performed with a preliminary visual analysis followed by the removal of artifacts. The scientists assessed the level of representation of the main alpha rhythm and its spatial distribution. The ECG registration was in the first lead. The duration of EEG-ECG registration is about 15 minutes in a typical sequence of stress tests: baseline, rhythmic photostimulation, hyperventilation with recovery periods. The spectral analysis of the EEG‒ECG was performed automatically on a Neurotravel device (Italy). The authors found lower alpha and delta activity in the initial EEG spectra in older drivers aged 45–66 years, which contributed to the cumulative effect of stress load. Light stress stimulation significantly reduced alpha and delta activity and increased beta EEG activity in both groups, while in the older group the high-frequency range significantly exceeded the low-frequency range. Simultaneously with the changes in the EEG, stress photostimulation increased the energy of the ECG spectra in all frequency ranges. The authors established the appearance of additional stress indicators during light stimulation in older drivers based on the correlation of EEG‒ECG spectra (alpha/delta) and the absence of such changes in the group of young drivers aged 25–35 years. Moreover, the EEG–ECG correlation coefficients and their critical values increased in groups in increments of 6 years — from 35 to 65 years. Elderly drivers, based on neuro-cardiodynamic indicators, were in a state of chronic stress and the resulting risks to health and professional activity, and the identified physiological mechanisms of stress may be at the heart of ways to prevent it. Limitations. The study is limited to the assessment of EEG and ECG indicators in 300 heavy truck drivers. Ethics. The study was performed using non-invasive methods and approved by the Bioethical Committee of the Research Institute of Complex Problems of Hygiene and Occupational Diseases, Protocol No. 5, § 1 dated 12/26/2018, in accordance with the Helsinki Declaration of the World Medical Association "Ethical Principles of conducting medical research with human participation as a subject" as amended in 2013 and the "Rules of Good Clinical Practice" approved by by Order of the Ministry of Health of the Russian Federation No. 200n dated April 1, 2016. All the examined patients signed a voluntary informed consent to participate in the study.

Duration of onset, body temperature and C-reactive protein can be used to predict the results of pus culture in children with acute osteomyelitis of long bones

BackgroundWith the application of PCR testing and Metagenomic Next-Generation Sequencing(mNGS), the detection rate of causative organisms in paediatric bone and joint infections has been greatly improved. The aim of our study is to identify some indicators that could be used to distinguish the culture results to optimize the use of PCR and mNGS.MethodsIn this study, a total of 117 cases of acute osteomyelitis of long bones in children who underwent pus culture were included. Patients were grouped as culture-negative (n:21) and culture-positive (n:96) groups according to the results of pus culture. Age, sex, duration of onset, maximum body temperature at onset, inflammatory indicators and D-dimer after admission were systematically collected for all patients and were compared for both groups. ROC curve (ROC) was used to evaluate the diagnostic efficiency of culture-negative. Logistic regression analysis was conducted to determine independent risk factors for culture-negative.ResultsThere was no significant difference in age, sex and erythrocyte sedimentation rate between culture-negative group and culture-positive group (P > 0.05). The duration of onset was longer, and the temperature, white blood cells, neutrophils count, C-reactive protein and D-dimer were less elevated in culture-negative acute osteomyelitis (P < 0.05). Duration of onset, maximum body temperature at onset, white blood cell count, neutrophil count, C-reactive protein, and D-dimer have certain diagnostic efficacy in judging the efficacy of negative culture. Logistic regression analysis indicated that the duration of onset more than 6.5 days, the maximum body temperature at onset lower than 38.35℃ and C-reactive protein lower than 78.40 mg/L were independent risk factors for negative culture (P < 0.05).ConclusionsOur study revealed that duration of onset more than 6.5 days, maximum body temperature at onset lower than 38.35℃ and C-reactive protein lower than 78.40 mg/L were independent risk factors for predicting negative culture. In children with this type of acute osteomyelitis, we recommend that the pus be tested by PCR or mNGS as a priority.

Recognition of psychoneurological state of children with autism spectrum disorder by features of speech: acoustic and perceptual features

The work is devoted to studying of recognition of psychoneurological state of children aged 5–14 with autism spectrum disorder (ASD), n = 35, and typically developing (TD) children, n = 45, by adults. A perceptual analysis was carried out, in which adult native speakers of the Russian language (auditors), n = 206, took part. Test sequences (audiotests) with words and phrases of ASD and TD children selected from spontaneous speech recordings were created for perceptual research. Auditors’ task was to determine psychoneurological state of children (typical/atypical development) based on auditory perception. A spectrographic analysis of phrases of children was carried out. Phrases of ASD children in comparison with TD children are characterized by lower speech rate, fewer words, great stressed and unstressed vowel duration, great values of pitch in phrase, word, stressed and unstressed vowel.

Genetic variants in childhood-onset dilatation of thoracic aorta in a consanguineous population

IntroductionChildhood-onset thoracic aortic dilatation (TAD) is a heterogenous group of genetic conditions the inheritance of which is largely dominant. To our knowledge, the influence of consanguinity on childhood-onset TAD has not been addressed systematically. We aim to study a cohort of children with TAD in our highly consanguineous population. MethodsChildren with TAD were consecutively recruited. Based on the likelihood of a founder mutation, genetic test was categorized to either targeted gene testing or multi-gene sequencing, followed by genetic screening of first-degree relatives. Clinical data and outcome were reviewed. ResultsThirty-three children, from 20 families, had childhood-onset TAD. Genetic test was positive in 20 children, from 13 families, providing a yield of 65 % (13/20). The median age of onset of TAD was 4.5 years. Eight variants were detected in 4 genes (FBN1, EFEMP2, ACTA2, KANSL1) with a homozygous EFEMP2 variant found in 6 families (46.2 %). Surgical intervention was required in 14 (70 %) cases (13 with EFEMP2, 1 with FBN1 variants) at a median age of 3.5 years. All patients are alive (ages range:3–31 years). ConclusionsOur work illustrates the impact of consanguinity on the genetics of childhood-onset TAD elucidating severe presentation of recessively inherited form. Our data underscores the importance of genetic screening and early recognition of TAD to achieve excellent outcome.

A comprehensive epidemiological approach documenting an outbreak of H5N1 highly pathogenic avian influenza virus clade 2.3.4.4b among gulls, terns, and harbor seals in the Northeastern Pacific

Highly pathogenic avian influenza viruses (HPAIV) H5N1 clade 2.3.4.4b continue to have unprecedented global impacts on wild birds and mammals, with especially significant mortality observed in colonial surface-nesting seabirds and in some marine mammal species. In July of 2023 H5N1 HPAIV 2.3.4.4b was detected in Caspian terns nesting on Rat Island, Washington USA. An estimated 1,800–1,900 adult terns populated the breeding colony, based on aerial photographs taken at the start of the outbreak. On a near-weekly basis throughout July and August, we counted and removed carcasses, euthanized moribund birds, and collected swab and tissue samples for diagnostic testing and next-generation sequencing. We directly counted 1,101 dead Caspian tern adults and 520 dead chicks, indicating a minimum 56% loss of the adult colony population and potential impacts to reproductive success. Combining the observed mortality on Rat Island with HPAI-related Caspian tern deaths recorded elsewhere in Washington and Oregon, we estimate that 10–14% of the Pacific Flyway population was lost in the summer of 2023. Comparatively few adult Glaucous-winged gulls (hybrids) nesting on Rat Island died (~3% of the local population), although gull chick mortality was high. Sixteen harbor seals in the immediate or nearby area stranded during the outbreak, and H5N1 HPAIV was detected in brain and/or lung tissue of five seals. These cases are the first known detections of HPAIV in a marine mammal on the Pacific coast of North America. Phylogenetic analyses support the occurrence of at least three independent avian-mammalian virus spillover events (tern or gull to harbor seal). Whole genome sequencing indicated that H5N1 HPAIV may have been introduced to Washington from Caspian terns in Oregon. Ongoing monitoring and surveillance for H5N1 HPAIV in the marine environment is necessary to understand the epidemiology of this virus, assess conservation impacts to susceptible species, and provide support for data-driven management and response actions.

Analysis of the composition and function of rhizosphere microbial communities in plants with tobacco bacterial wilt disease and healthy plants

ABSTRACT To explore the factors influencing the occurrence of bacterial wilt, the differences in the physicochemical properties, microbial community composition and function between rhizosphere soil of tobacco plants with bacterial wilt and healthy plants in the tobacco planting area of Fuzhou City, Jiangxi Province were analyzed and compared. The results showed that the rhizosphere soil of diseased tobacco exhibited significantly reduced levels of exchangeable potassium, water-soluble potassium, nitrate nitrogen, total nitrogen and pH, in comparison to the rhizosphere soil of healthy plants. Conversely, the available phosphorus content of the rhizosphere soil of diseased tobacco was significantly increased. The amount of Ralstonia solanacearum in soil was negatively correlated with pH, nitrate nitrogen and total nitrogen, and positively correlated with exchangeable potassium and water-soluble potassium. A total of 43 genera were significantly different between the two groups of rhizosphere soil, of which 24 genera were enriched in the rhizosphere of healthy plants, including Ideonella , Rhizophagus , Rhizobacter , Altererythrobacter and Ignavibacterium associated with plant disease resistance, Thermodesulfovibrio , Syntrophorhabdus , Syntrophus , Chlorobium , Hydrogenophaga and Limnohabitans associated with soil sulfur metabolism, as well as Ignavibacterium , Ideonella , Derxia and Azohydromonas associated with soil nitrogen cycling. Kyoto Encyclopedia of Genes and Genomes functional analysis of the unigenes obtained by metagenomic sequencing also showed that the differential unigenes were significantly enriched in the sulfur metabolism pathway. In addition, the rhizosphere soil of diseased tobacco plants exhibited a higher abundance of antibiotic-producing actinomycetes and an increased load of antibiotic resistance genes compared to that of healthy plants. In general, lower pH value, less content of nitrate nitrogen and total nitrogen, and more content of exchangeable potassium and water-soluble potassium could contribute to onset of bacterial wilt. Twenty-four genera, including Ideonella and Rhizophagus , may construct a healthy microecological network in the rhizosphere of tobacco plants. All these factors may interact with each other to control the development of bacterial wilt. This complicated interaction network needs to be explored further. IMPORTANCE Previous studies have mainly focused on the differences in microbial species composition between healthy and diseased soils, but the differences in microbial community functions between two types of soil have not been well characterized. In this study, soil samples in diseased and healthy plant rhizospheres were collected for physicochemical property testing and metagenomic sequencing. We focused on analyzing the differences in physicochemical properties and microbial community functions between these soils, as well as the correlation between these factors and pathogen content. The results of this study provide a theoretical basis for further understanding the occurrence of tobacco bacterial wilt in the field.

Fast‐MedNeXt: Accelerating the MedNeXt Architecture to Improve Brain Tumour Segmentation Efficiency

ABSTRACTWith the rapid development of medical imaging technology, 3D image segmentation technology has gradually become a mainstream method, especially in brain tumour detection and diagnosis showing its unique advantages. The technique makes full use of 3D spatial information to locate and analyze tumours more accurately, thus playing an important role in improving diagnostic accuracy, optimising treatment planning and promoting research. However, it also suffers from significant computational expenditure and delayed processing pace. In this paper, we propose an innovative optimisation scheme to address this problem. We thoroughly investigate the MedNeXt network and propose Fast‐MedNeXt, which aims to increase the processing speed while maintaining accuracy. First, we introduce the partial convolution (PConv) technique, which replaces the deep convolutional layers in the network. This improvement effectively reduces computation and memory requirements while maintaining efficient feature extraction. Second, based on PConv, we propose PConv‐Down and PConv‐Up modules, which are applied to the up‐sampling and down‐sampling modules to further optimise the network structure and improve efficiency. To confirm the efficacy of the approach, we carried out a sequence of tests in the multimodal brain tumour segmentation challenge 2021 (BraTS2021). By comparing with the MedNeXt series network, the Fast‐MedNeXt reduced the latency by 22.1%, 20.5%, 15.8%, and 11.4% respectively, while the average accuracy also increased by 0.475% and 0.2% respectively. These significant performance improvements demonstrate the effectiveness of Fast‐MedNeXt in 3D medical image segmentation tasks and provide a new and more efficient solution for the field.

Clinical characteristics and prognosis of bloodstream infections with carbapenem-resistant Gram-negative organisms in patients with hematological malignancies: A multicenter case-control study in China

To investigate clinical characteristics of hematological malignancy (HM) patients with carbapenem-resistant gram-negative organism (CRO) bloodstream infections (BSI) in China, and to elucidate the prognostic risk factors of CRO BSI. We conducted a multicenter case-control study of 201 HM patients with CRO BSI between 2018-2020. Antimicrobial susceptibility testing and whole genome sequencing were performed for CRO isolates. Independent risk factors for 28-day crude mortality of were analyzed using Cox proportional hazards regression models. The subgroups of major species were also evaluated. The pathogens responsible for CRO BSI in HM patients dominated by ST11 CRKP, ST167 CREC and ST463 CRPA. Most isolates produced carbapenemases with KPC and NDM being the main. CRO isolates had resistance rates to conventional antimicrobials ranging from 55%-100% and poor susceptibility to novel antimicrobials related to carbapenemases and species. The 28-day crude mortality was 24.2%. Non-Hodgkin lymphoma, heart disease, blaKPC-2 positive, empirical antibiotic therapy with linezolid, Pitt bacteremia score >3.5 were risk factors for 28-day mortality and appropriate definitive antibiotic therapy, tigecycline-containing therapy and aminoglycoside-containing therapy were protective factors. blaKPC-2 positive in CRKP and ST463 in CRPA were associated with Pitt bacteremia score > 3.5. Solid tumor and other site infections before BSI were risk factors for ST463 CRPA BSI and Pulmonary infection before BSI was risk factor for KPC-KP BSI. The antimicrobial resistance of CRO isolates for BSI in HM patients is critical. HM patients with CRO BSI should be treated with appropriate definitive antibiotic therapy based on early clarification of pathology and their antimicrobial susceptibility.

Molecular identification and antimicrobial resistance patterns of enterobacterales in community urinary tract infections among indigenous women in Ecuador: addressing microbiological misidentification

BackgroundAntibiotic resistance of Enterobacterales poses a major challenge in the treatment of urinary tract infections (UTIs). In low- and middle-income countries (LMICs), standard microbiological (i.e. urine culture and simple disk diffusion test) methods are considered the “gold standard” for bacterial identification and drug susceptibility testing, while PCR and DNA sequencing are less commonly used. In this study, we aimed to re-identifying Enterobacterales as the primary bacterial agents responsible for urinary tract infections (UTIs) by comparing the sensitivity and specificity of traditional microbiological methods with advanced molecular techniques for the detection of uropathogens in indigenous women from Otavalo, Ecuador.MethodsA facility-based cross-sectional study was conducted from October 2021 to February 2022 among Kichwa-Otavalo women. Pathogens from urine samples were identified using culture and biochemical typing. Morphological identification was doble-checked through PCR and DNA sequencing of 16S, recA, and rpoB molecular barcodes. The isolates were subjected to antimicrobial susceptibility-testing using disk diffusion test.ResultsThis study highlighted a 32% misidentification rate between biochemical and molecular identification. Using traditional methods, E. coli was 26.19% underrepresented meanwhile Klebsiella oxytoca was overrepresented by 92.86%. Furthermore, the genera Pseudomonas, Proteus, and Serratia were confirmed to be E. coli and Klebsiella spp. by molecular method, and one Klebsiella spp. was reidentified as Enterobacter spp. The susceptibility profile showed that 59% of the isolates were multidrug resistant strains and 31% produced extended spectrum beta-lactamases (ESBLs). Co-trimoxazole was the least effective antibiotic with 61% of the isolates resistant. Compared to previous reports, resistance to nitrofurantoin and fosfomycin showed an increase in resistance by 25% and 15%, respectively.ConclusionsCommunity-acquired UTIs in indigenous women in Otavalo were primarily caused by E. coli and Klebsiella spp. Molecular identification (16S/rpoB/recA) revealed a high rate of misidentification by standard biochemical and microbiological techniques, which could lead to incorrect antibiotic prescriptions. UTI isolates in this population displayed higher levels of resistance to commonly used antibiotics compared with non-indigenous groups. Accurate identification of pathogens causing UTIs and their antibiotic susceptibility in local populations is important for local antibiotic prescribing guidelines.

Antimicrobial resistance in plant endophytes associated with poultry-manure application revealed by selective culture and whole genome sequencing

Poultry manure is widely used as organic fertilizer in agriculture during the cultivation of crops, but the persistent high-level use of antibiotics in poultry production has raised concerns about the selection for reservoirs of antimicrobial resistance genes (ARGs). Previous studies have shown that the addition of poultry manure can increase the abundance of genes associated with resistance to tetracyclines, aminoglycosides, fluoroquinolones, sulfonamides, bacitracin, chloramphenicol, and macrolide-lincosamide-streptogramin in soil and plants. Understanding the microbial populations that harbor these ARGs is important to identify microorganisms that could enter the human food chain. Here, we test the hypothesis that environmental exposure to poultry manure increases the occurrence of antimicrobial resistance (AMR) in plant endophytes using selective culture, phenotypic Antibiotic Susceptibility Testing (AST), phylogenetic analysis, and whole genome sequencing (WGS). Endophytes from poultry manure treated Sorghum bicolor (L.) Moench plant root and stem samples showed increased phenotypic and genotypic resistance against multiple antibiotics compared to untreated controls. Comparison of AMR phenotype-to-genotype relationships highlighted the detection of multi-drug resistant (MDR) plant endophytes, demonstrating the value of genomic surveillance for emerging drug-resistant pathogens. The increased occurrence of ARGs in poultry manure-exposed endophytes highlights the need for responsible antibiotic use in poultry and animal farming to reduce contamination of ecological niches and transgression into endophytic plant microbiome compartments. It also emphasizes the requirement for proper manure management practices and vigilance in monitoring and surveillance efforts to tackle the growing problem of antibiotic resistance and preserve the efficacy of antibiotics for human and veterinary medicine.

B-074 Comparative Evaluation of Clinical Utility between Indirect Immunofluorescence and Immunoassay Methods for Antineutrophil Cytopalsmic Antibody Detection

Abstract Background ANCA testing is essential for the diagnosis of vasculitis, commonly using indirect immunofluorescence(IIF) and immunoassay(IA) methods. Recent guidelines recommend IA as the primary test for suspected GPA and MPA, but actual practice varies among physicians. Here, we retrospectively analyzed ANCA test results, focusing on new cases of ANCA-associated vasculitis, and compared the utility of IIF and IA to guide appropriate test selection. Methods From January 2022 to July 2023, 24 new cases of ANCA-associated vasculitis were diagnosed. We checked whether ANCA testing was performed and the results, and compared the turnaround time(TAT) and costs. IIF ANCA tests were performed on a computer-based automated instrument(Helios), and IA ANCA tests were performed on automated instruments(Alegria, Phadia 250) with MPO antigen and PR3 antigen tested simultaneously in a bundled prescription. Results Among the 24 cases, there were 6 GPA, 7 MPA, 7 EGPA, and 4 ANCA-associated GN, with varying diagnostic sensitivities. GPA and EGPA had different ANCA positivity rates than previously reported, suggesting potential ethnic variations in ANCA expression and positivity rates. The test sequence and results, summarized in Table 1, showed 21 out of 22 cases had identical concordance between methods, with an excellent concordance rate (kappa value=0.83). TAT for IIF was 3,452 minutes, and for IA was 1,491 minutes for the whole period, but IA TAT increased to 2,748 minutes after implementing a days-of-the-week testing system. Test costs, with a relative value score of 118.99 for IIF and 156.05 for IA, indicated IA cost approximately 2.6 times higher, considering the bundled prescription of both antigens for IA ANCA testing. Conclusions Evaluating the utility of both methods for ANCA detection, our study highlights high inter-method agreement in ANCA-associated vasculitis. Considering overall cost and turnaround time, IIF ANCA testing remains effective as a screening test, demonstrating standalone effectiveness.

B-035 Atellica CI Analyzer Sample-to-sample Carryover Performance Evaluation

Abstract Background Laboratories in many parts of the world are required to demonstrate/confirm the lack of sample carryover on their analyzer under their laboratory conditions using patient populations. Sample carryover occurs when analyte adheres to the probe and/or cuvette and contaminates a subsequent sample or cuvette in the testing sequence of the analyzer. Inadequate washing of the sample aspiration/ dispensing device and cuvette can cause this type of carryover. The Atellica® CI Analyzer uses independent integration and random-access sampling to process chemistry (CH) and immunoassay (IM) tests. The immunoassays performed on the IM portion of the analyzer utilizes a single-use, disposable sample tip and reaction cuvette which reduces sample carryover from IM to CH tests, whereas a reusable probe is used for CH assays—which is washed between sampling. This study evaluated the potential for carryover of CH samples from one sample to the next due to washing protocols on the Atellica® CI Analyzer in four independent real world clinical laboratories. Methods Carryover verification was performed as per CLSI GP31-A:2009 and Vericheck protocols. The study was conducted at four sites (UAE, France, Croatia, U.S.) using 2 serum samples prepared in 12 sample containers-9 for low (L) and 3 for high (H) concentrations of analyte. For glucose (GluH_3), UAE, L and H were 63 and 356 mg/dL (3.5 and 19.8 mmol/L), Croatia, 77.4 and 237.5 mg/dL (4.3 and 12.2 mmol/L), and U.S., 101 and 374 mg/dL (5.6 and 20.8 mmol/L). For creatine kinase (CK_L), France, L and H were 40 and 1400 U/L. Samples were tested in a defined sequence: L1, L2, L3, H1, L4, H2, L5, H3, L6, L7, L8, L9 and results analyzed for % carryover. Carryover was defined as average of L4, L5, L6 minus the average of L1, L2, L3, L7, L8, L9 divided by the average of H1, H2, H3. Results No significant carryover was observed at any site. Carryover % for UAE, France, Croatia, and U.S was 0.500%, -0.085%, -0.095%, and -0.095%, respectively. Conclusions When using high positive samples at concentrations at or greater than the upper limit of the clinical range, negligible CH sample carryover was found from the Atellica CI Analyzers in four independent real world clinical laboratory settings.

Rapid Access Diagnostics for Asthma (RADicA): protocol for a prospective cohort study to determine the optimum series of investigations to diagnose asthma using conventional and novel tests

IntroductionThe diagnosis of asthma is often based on characteristic patterns of symptoms in the absence of an alternative explanation, resulting in over and under diagnosis. Therefore, diagnostic guidelines usually recommend...

Evaluating culture-free targeted next-generation sequencing for diagnosing drug-resistant tuberculosis: a multicentre clinical study of two end-to-end commercial workflows.

Drug-resistant tuberculosis remains a major obstacle in ending the global tuberculosis epidemic. Deployment of molecular tools for comprehensive drug resistance profiling is imperative for successful detection and characterisation of tuberculosis drug resistance. We aimed to assess the diagnostic accuracy of a new class of molecular diagnostics for drug-resistant tuberculosis. We conducted a prospective, cross-sectional, multicentre clinical evaluation of the performance of two targeted next-generation sequencing (tNGS) assays for drug-resistant tuberculosis at reference laboratories in three countries (Georgia, India, and South Africa) to assess diagnostic accuracy and index test failure rates. Eligible participants were aged 18 years or older, with molecularly confirmed pulmonary tuberculosis, and at risk for rifampicin-resistant tuberculosis. Sensitivity and specificity for both tNGS index tests (GenoScreen Deeplex Myc-TB and Oxford Nanopore Technologies [ONT] Tuberculosis Drug Resistance Test) were calculated for rifampicin, isoniazid, fluoroquinolones (moxifloxacin, levofloxacin), second line-injectables (amikacin, kanamycin, capreomycin), pyrazinamide, bedaquiline, linezolid, clofazimine, ethambutol, and streptomycin against a composite reference standard of phenotypic drug susceptibility testing and whole-genome sequencing. Between April 1, 2021, and June 30, 2022, 832 individuals were invited to participate in the study, of whom 720 were included in the final analysis (212, 376, and 132 participants in Georgia, India, and South Africa, respectively). Of 720 clinical sediment samples evaluated, 658 (91%) and 684 (95%) produced complete or partial results on the GenoScreen and ONT tNGS workflows, respectively, with 593 (96%) and 603 (98%) of 616 smear-positive samples producing tNGS sequence data. Both workflows had sensitivities and specificities of more than 95% for rifampicin and isoniazid, and high accuracy for fluoroquinolones (sensitivity approximately ≥94%) and second line-injectables (sensitivity 80%) compared with the composite reference standard. Importantly, these assays also detected mutations associated with resistance to critical new and repurposed drugs (bedaquiline, linezolid) not currently detectable by any other WHO-recommended rapid diagnostics on the market. We note that the current format of assays have low sensitivity (≤50%) for linezolid and more work on mutations associated with drug resistance is needed. This multicentre evaluation demonstrates that culture-free tNGS can provide accurate sequencing results for detection and characterisation of drug resistance from Mycobacterium tuberculosis clinical sediment samples for timely, comprehensive profiling of drug-resistant tuberculosis. Unitaid.

Utilizing non-invasive prenatal test sequencing data for human genetic investigation

Non-invasive prenatal testing (NIPT) employs ultra-low-pass sequencing of maternal plasma cell-free DNA to detect fetal trisomy. Its global adoption has established NIPT as a large human genetic resource for exploring genetic variations and their associations with phenotypes. Here, we present methods for analyzing large-scale, low-depth NIPT data, including customized algorithms and software for genetic variant detection, genotype imputation, family relatedness, population structure inference, and genome-wide association analysis of maternal genomes. Our results demonstrate accurate allele frequency estimation and high genotype imputation accuracy (R2>0.84) for NIPT sequencing depths from 0.1× to 0.3×. We also achieve effective classification of duplicates and first-degree relatives, along with robust principal-component analysis. Additionally, we obtain an R2>0.81 for estimating genetic effect sizes across genotyping and sequencing platforms with adequate sample sizes. These methods offer a robust theoretical and practical foundation for utilizing NIPT data in medical genetic research.

Effects of forward head posture on the centre of pressure sway during dual-task balancing with smartphone engagement

PurposeForward head posture (FHP) may affect postural stability and balance. This study examines how smartphone texting while balancing, as a dual task, influences the body’s centre of pressure (CoP) during single-leg standing in individuals with FHP compared to those with normal posture.MethodsThe study utilized a randomized repeated measures design to assess single-leg standing balance and CoP across six conditions. Fifty participants, divided into two groups (25 with FHP and 25 with normal posture), underwent testing sequences involving non-smartphone usage (NU; single task), one-handed smartphone usage (1H; dual task), and two-handed smartphone usage (2H; dual task) on both soft and hard surfaces.ResultsThe study found that individuals with FHP showed increased CoP sway compared to those with normal posture across all conditions. Both 1H and 2H increased sway compared to non-usage for both groups, with a notable difference in FHP individuals on soft surfaces, where 2H led to greater sway.ConclusionsIndividuals with FHP are at a higher risk of balance issues during both NU and smartphone use, particularly during 2H. These results highlight the importance of addressing the impact of FHP on balance control and implementing interventions to reduce associated risks.

Synergistic antimicrobial activity of alpha-linolenic acid in combination with tetracycline or florfenicol against multidrug-resistant Salmonella typhimurium

Salmonella is a major foodborne pathogen that can be transmitted from livestock and poultry to humans through the food chain. Due to the widespread use of antibiotics, antibiotic resistance Salmonella has become an important factor threatening food safety. Combining antibiotic and non-antibiotic agents is a promising approach to address the widespread emergence of antibiotic-resistant pathogens.In this study, we investigated the antibiotic resistance profile and molecular characterization of different serotypes of Salmonella isolated from large-scale egg farms using drug susceptibility testing and whole genome sequencing. The synergistic effect of alpha-linolenic acid (ALA) with antibiotics was evaluated using the checkerboard test and time-kill curve. The molecular mechanism of α-linolenic acid synergism was explored using biochemical assays, pull-down assays, and molecular docking. In vivo efficacy of ALA in combination with florfenicol (FFC) or tetracycline (TET) against multidrug-resistant (MDR) Salmonella enterica subsp. enterica serovar typhimurium was also investigated using a mouse model.We found that ALA reduced the minimum inhibitory concentration (MIC) of tetracycline and florfenicol in all strains tested. When ALA (512 mg/L) was combined with florfenicol (32 mg/L) or tetracycline (16 mg/L), we observed disruption of cell membrane integrity, increased outer membrane permeability, lowered cell membrane potential, and inhibition of proton-drive-dependent efflux pumps. The synergistic treatment also inhibited biofilm production and promoted oxidative damage. These changes together led to an increase in bacterial antibiotic susceptibility. The improved efficacy of ALA combination treatment with antibiotics was validated in the mouse model. Molecular docking results indicate that ALA can bind to membrane proteins via hydrogen bonding. Our findings demonstrated that combined treatment using ALA and antibiotics is effective in preventing infections involving MDR bacteria. Our results are of great significance for the scientific and effective prevention and control of antibiotic resistance Salmonella, as well as ensuring food safety.

Investigation and successful control of a Candida auris outbreak at a tertiary health care facility in Kenya

ObjectivesThis study aimed to conduct an epidemiological and genomic investigation of a hospital outbreak of Candida auris, and implement measures for its control. MethodsWe collected demographic and clinical data from medical records of patients with C. auris from January 2017 to June 2019 after identifying increased cases in April 2019. Point-prevalence surveys for C. auris colonisation were conducted in the critical care units (CCU). Antifungal susceptibility testing and genomic sequencing of isolates were performed. A bundle of infection prevention and control measures was instituted. ResultsThirty-two patients with C. auris were identified. All patients had a history of CCU admission. A total of 283 screening swabs were obtained and 57 isolates of C. auris identified. Antifungal susceptibility testing was performed on 48 isolates. All but two isolates were resistant to fluconazole; one isolate was also resistant to amphotericin B. Forty-one of 46 isolate genomes were clonally related and formed a distinct genetic cluster in Clade III. C. auris colonisation reduced from 42% in June 2019 to 1% in August 2019, and no new hospital-acquired colonisation was identified in the subsequent 9 months. ConclusionsWe identified a new genetic subcluster of Clade III C. auris. We also show that strict implementation of infection prevention measures can lead to substantial reductions in C. auris transmission.

Physiological Profile Assessment and Self-Measurement of Healthy Students through Remote Protocol during COVID-19 Lockdown.

Background: The COVID-19 pandemic has led to reduced physical activity and increased sedentary behaviors, negatively impacting mental and physical health. Engaging in physical activity at home during quarantine became essential to counteracting these adverse effects. To develop appropriate physical activity programs, assessing individuals' fitness levels and the impact of inactivity is crucial. This study aims to compare motor abilities-including flexibility, balance, reaction time, cardiovascular endurance, and lower and upper limb strength-assessed both in-person and remotely, to determine the accuracy and repeatability of self-administered tests. Methods: A total of 35 young subjects (age 24.2 ± 1.97 years, BMI 22.4 ± 2.61 kg/m2) participated in this study. Each participant underwent a battery of motor ability tests designed to assess various fitness components. The tests were administered twice for each subject: once in a laboratory setting and once remotely at home. The sequence of tests was randomly assigned to ensure unbiased results. Both the in-person and remote assessments were used to evaluate the accuracy and reliability of self-administered tests. Results: The comparison of test results between the laboratory and remote settings revealed percentage differences ranging from 5% to 10%. This variation is considered an acceptable margin of error, suggesting that the tests conducted remotely were relatively accurate when compared to those performed in a controlled laboratory environment. Conclusions: The findings indicate that remote fitness testing is a promising method for evaluating motor abilities. With an acceptable margin of error, remote assessments can be effectively used to personalize training programs based on individuals' physiological characteristics. This approach may be particularly beneficial during times of limited access to fitness facilities, such as during quarantine, or for individuals seeking more flexible fitness evaluation methods.

Dynamic behavior of kaolin clay under bidirectional cyclic loading for suction bucket foundation

The stability of offshore wind turbine (OWT) foundations is largely influenced by the dynamic characteristics of marine clay. In offshore engineering, marine clays may exposed to bidirectional cyclic stresses. To empirically examine the influence of different cyclic stress magnitudes on the dynamic characteristics of marine clay at different depth, a sequence of dynamic triaxial tests is conducted on marine kaolin clay, encompassing diverse values of the cyclic stress ratio (CSR) and effective confining pressure (p'0). Some valuable inferences have been drawn: At low cyclic stress ratios (CSR≤0.3), the axial strain (εa), stiffness (k), and excess pore water pressure (EPWP) of the specimen exhibit logarithmic increase, logarithmic decrease, and logarithmic accumulation, respectively. Under high cyclic stress ratio conditions (CSR＞0.3), the axial strain increases linearly, the stiffness deteriorates linearly, and the logarithmic excess pore water pressure accumulates. After analyzing the dynamic characteristics data of kaolin clay, the correlation between excess pore water pressure and axial strain was established. By analyzing the vertical cyclic mechanism of suction bucket foundation, a prediction model for suction bucket foundation under vertical cyclic loading was established based on the dynamic characteristics of marine clay and the model was validated. The comparison between the validation outcomes and the experimental outcomes shows good agreement. This study provides certain guidance for the design of suction bucket foundations for OWTs.