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Data Description: This dataset comprises fluorescence micrographs of HeLa cells, specifically labelled to identify nuclei and cell cytoplasm. These images were acquired as a technical calibration for a high-content screening study detailed and published in [1]. The HeLa cell line (ATCC-CCL-2), a widely used immortalised cell line in laboratory research, was cultured under standard conditions. Post-cultivation, the cells were fixed and stained with fluorescent dyes to visualise the nuclei and cytoplasm. The nuclei were stained with DAPI (4',6-diamidino-2-phenylindole), a blue-fluorescent DNA stain, while fluorescent-labeled phalloidin was used to detect actin filaments and delineate the cytoplasm. The entire process of cell culture, fixation, staining, and imaging adhered strictly to the protocols described in [1]. The preprocessed dataset includes 2,676 8-bit RGB images, each with a pixel resolution of 520 x 696 pixels. In these images, only two of the RGB channels are utilized: the red channel represents the cytoplasm, and the blue channel represents the nuclei. The dataset is divided into training, validation, and test subsets in a 70:20:10 ratio. The entire dataset is accompanied by instance segmentation masks for nuclei and cytoplasm objects obtained through a specialised CellProfiler [2] software. Notably, the test subset was annotated manually by a specialist, ensuring high-quality annotations. The original raw images are of a higher resolution, 1040 x 1392 pixels, and have a bit depth of 16 bits, providing more detailed information for advanced analyses. File Description: The file structure of the zip files is as follows: HeLaCytoNuc_{train/validation/test}.zip -> - images -> {filename}.tif - nuclei_masks  -> {filename}.tif - cytoplasm_masks  -> {filename}.tif HeLaCytoNuc_raw_images.zip -> {filename}.tif HeLa

Data Description The VACVPlaque dataset comprises spatially correlated objects, specifically virological plaques, which are circular phenotypes indicative of vaccinia virus (VACV) spread, and the wells of the assay plate. The virus plaque assay is a common method performed by infecting a monolayer of host cells (indicator cells) that are grown in the wells of assay plates or dishes. The host cells are infected with varying concentrations of a highly diluted virus inoculum. After an incubation period, typically around 48 hours, the cells are fixed with formaldehyde and stained with a dye to reveal the plaques or areas of cell death. By counting these plaques, researchers can calculate the number of infectious particles present in the original inoculum as described in [1]. This dataset consists of mobile photographs of 6-well tissue culture plates where the VACV plaque assay was conducted. The photographs were taken using two different mobile phones, resulting in 211, 16-bit RGB images with a resolution of 2448 x 3264 pixels. Each plate was photographed from two different perspectives using two different devices, meaning there are two images of the same plate but from different angles and devices. To aid in the training of machine learning models, the dataset is divided into training, validation, and test subsets in a 70:20:10 ratio. To prevent data leaks, only one perspective of each image is included in the validation and test subsets. The training subset, which includes images from both perspectives, consists of 148 images. File Description: VACVPlaque_train.zip -> train holdout VACVPlaque_validation.zip -> validation holdout VACVPlaque_test.zip -> test holdout Each zip file contains: images -> {filename}.tif plaque_masks -> {filename}.tif well_masks -> {filename}.tif References: 1. Dulbecco, Re

01 februari 2023, Item does not contain fulltext, To assess and describe the aetiology and management of febrile illness in children with primary or acquired immunodeficiency at high risk of serious bacterial infection, as seen in emergency departments in tertiary hospitals. Prospective data on demographics, presenting features, investigations, microbiology, management, and outcome of patients within the 'Biomarker Validation in HR patients' database in PERFORM, were analysed. Immunocompromised children (< 18 years old) presented to fifteen European hospitals in nine countries, and one Gambian hospital, with fever or suspected infection and clinical indication for blood investigations. Febrile episodes were assigned clinical phenotypes using the validated PERFORM algorithm. Logistic regression was used to assess the effect size of predictive features of proven/presumed bacterial or viral infection. A total of 599 episodes in 482 children were analysed. Seventy-eight episodes (13.0%) were definite bacterial, 67 episodes probable bacterial (11.2%), and 29 bacterial syndrome (4.8%). Fifty-five were definite viral (9.2%), 49 probable viral (8.2%), and 23 viral syndrome (3.8%). One hundred ninety were unknown bacterial or viral infections (31.7%), and 108 had inflammatory or other non-infectious causes of fever (18.1%). Predictive features of proven/presumed bacterial infection were ill appearance (OR 3.1 (95% CI 2.1-4.6)) and HIV (OR 10.4 (95% CI 2.0-54.4)). Ill appearance reduced the odds of having a proven/presumed viral infection (OR 0.5 (95% CI 0.3-0.9)). A total of 82.1% had new empirical antibiotics started on admission (N = 492); 94.3% proven/presumed bacterial (N = 164), 66.1% proven/presumed viral (N = 84), and 93.2% unknown bacterial or viral infections (N = 177). Mortality was 1.9% (N = 11) and 87.1% made full recovery (N = 522). Conclusion: The aetiology of febrile illness in immunocompromised children is diverse. In one-third of cases, no cause for the fever will be identified. Justification for standard

01 september 2023, Contains fulltext : 295917.pdf (Publisher’s version ) (Open Access), BACKGROUND: The PERFORM study aimed to understand causes of febrile childhood illness by comparing molecular pathogen detection with current clinical practice. METHODS: Febrile children and controls were recruited on presentation to hospital in 9 European countries 2016-2020. Each child was assigned a standardized diagnostic category based on retrospective review of local clinical and microbiological data. Subsequently, centralised molecular tests (CMTs) for 19 respiratory and 27 blood pathogens were performed. FINDINGS: Of 4611 febrile children, 643 (14%) were classified as definite bacterial infection (DB), 491 (11%) as definite viral infection (DV), and 3477 (75%) had uncertain aetiology. 1061 controls without infection were recruited. CMTs detected blood bacteria more frequently in DB than DV cases for N. meningitidis (OR: 3.37, 95% CI: 1.92-5.99), S. pneumoniae (OR: 3.89, 95% CI: 2.07-7.59), Group A streptococcus (OR 2.73, 95% CI 1.13-6.09) and E. coli (OR 2.7, 95% CI 1.02-6.71). Respiratory viruses were more common in febrile children than controls, but only influenza A (OR 0.24, 95% CI 0.11-0.46), influenza B (OR 0.12, 95% CI 0.02-0.37) and RSV (OR 0.16, 95% CI: 0.06-0.36) were less common in DB than DV cases. Of 16 blood viruses, enterovirus (OR 0.43, 95% CI 0.23-0.72) and EBV (OR 0.71, 95% CI 0.56-0.90) were detected less often in DB than DV cases. Combined local diagnostics and CMTs respectively detected blood viruses and respiratory viruses in 360 (56%) and 161 (25%) of DB cases, and virus detection ruled-out bacterial infection poorly, with predictive values of 0.64 and 0.68 respectively. INTERPRETATION: Most febrile children cannot be conclusively defined as having bacterial or viral infection when molecular tests supplement conventional approaches. Viruses are detected in most patients with bacterial infections, and the clinical value of individual pathogen detection in determining treatment is low. New approaches are needed to help determine which feb

Contains fulltext : 294537.pdf (Publisher’s version ) (Open Access), BACKGROUND: To identify a diagnostic blood transcriptomic signature that distinguishes multisystem inflammatory syndrome in children (MIS-C) from Kawasaki disease (KD), bacterial infections, and viral infections. METHODS: Children presenting with MIS-C to participating hospitals in the United Kingdom and the European Union between April 2020 and April 2021 were prospectively recruited. Whole-blood RNA Sequencing was performed, contrasting the transcriptomes of children with MIS-C (n = 38) to those from children with KD (n = 136), definite bacterial (DB; n = 188) and viral infections (DV; n = 138). Genes significantly differentially expressed (SDE) between MIS-C and comparator groups were identified. Feature selection was used to identify genes that optimally distinguish MIS-C from other diseases, which were subsequently translated into RT-qPCR assays and evaluated in an independent validation set comprising MIS-C (n = 37), KD (n = 19), DB (n = 56), DV (n = 43), and COVID-19 (n = 39). RESULTS: In the discovery set, 5696 genes were SDE between MIS-C and combined comparator disease groups. Five genes were identified as potential MIS-C diagnostic biomarkers (HSPBAP1, VPS37C, TGFB1, MX2, and TRBV11-2), achieving an AUC of 96.8% (95% CI: 94.6%-98.9%) in the discovery set, and were translated into RT-qPCR assays. The RT-qPCR 5-gene signature achieved an AUC of 93.2% (95% CI: 88.3%-97.7%) in the independent validation set when distinguishing MIS-C from KD, DB, and DV. CONCLUSIONS: MIS-C can be distinguished from KD, DB, and DV groups using a 5-gene blood RNA expression signature. The small number of genes in the signature and good performance in both discovery and validation sets should enable the development of a diagnostic test for MIS-C.

Virological plaque assay is the primary way for the quantification of infectious virus particles in a virus suspension. It is performed by incubating a serial dilution of the virus inoculum with a monolayer of indicator cells. While some tools for automated quantification have been proposed in the past, they lack modularity and are often closed-source. PyPlaque is open-source software written in Python programming language for the analysis and quantification of plaque phenotypes that are observed in plaque assays by virologists. It is based on an abstract software architecture allowing to take into account glassware and specimen carriers and focus on phenotype-specific information. This, in turn, allows biologist to construct modular and simple program code that follows the logical flow of the experimental design and desired quantification outcomes. Furthermore, we demonstrate the comparability of PyPlaque to existing software and show how phenotype-based abstraction allows for seamless coding using Pythonic conventions.

Urinary tract infections (UTI) are a common disorder. Its diagnosis can be made by microscopic examination of voided urine for cellular markers of infection. We present a dataset containing 300 images and 3,562 manually annotated urinary cells labelled into seven classes of clinically significant urinary content. It is an enriched dataset with samples acquired from the unstained and untreated urine of patients with symptomatic UTI. The aim of the dataset is to facilitate UTI diagnosis in nearly all clinical settings by using a simple imaging system which leverages advanced machine learning techniques.  Data acquisition  300 urine samples were obtained from patients with symptomatic UTI between April and August 2022 from a specialist LUTS outpatient clinic in central London. Urine samples were collected as natural voids and processed on-site within one hour to mitigate cellular degradation. Brightfield microscopic examination (Olympus BX41F microscope frame, U-5RE quintuple nosepiece, U-LS30 LED illuminator, U-AC Abbe condenser) was performed at x20 objective (Olympus PLCN20x Plan C N Achromat 20x/0.4). A disposable haemocytometer (C Chip™) was used for enumeration of red cells (RBC), white cells (WBC), epithelial cells (EPC), and the presence of other cellular content per 1 µl of urine by two experienced microscopists. Images were acquired using the aforementioned brightfield microscope using a 0.5X C-mount adapter connected to a digital colour camera (Infinity 3S-1UR, Teledyne Lumenera). Images were taken in 16-bit colour in 1392 x 1040 .tif format using Capture and Analyse software. An enriched dataset approach was taken to maximise urinary cellular content in the acquired images. Such data curation was also necessary to overcome class imbalance. Daily Kohler illumination and global white balance was performed to ensure consistency in image acquisition.  Dataset annotation