Miracle-dz/UniFAR-FTUs huggingface dataset:
Stable Expression of Modified Green Fluorescent Protein in Group B Streptococci To Enable Visualization in Experimental Systems

Group B streptococcus (GBS) is a bacterium associated with various diseases in humans and animals. This study describes the development of a strategy to label and visualize GBS using a fluorescence biomarker, termed GFPmut3. We show that this biomarker can be successfully applied to track the growth of bacteria in liquid medium, and it enables the detailed visualization of GBS in the context of live human cells in real-time microscopic analysis. The system for GFPmut3 expression in GBS described in this study provides a new tool for the visualization of this organism in diverse research applications. ABSTRACT Group B streptococcus (GBS) is a Gram-positive bacterium associated with various diseases in humans and animals. Many studies have examined GBS physiology, virulence, and microbe-host interactions using diverse imaging approaches, including fluorescence microscopy. Strategies to label and visualize GBS using fluorescence biomarkers have been limited to antibody-based methods or nonspecific stains that bind DNA or protein; an effective plasmid-based system to label GBS with a fluorescence biomarker would represent a useful visualization tool. In this study, we developed and validated a green fluorescent protein (GFP)-variant-expressing plasmid, pGU2664, which can be applied as a marker to visualize GBS in experimental studies. The synthetic constitutively active CP25 promoter drives strong and stable expression of the GFPmut3 biomarker in GBS strains carrying pGU2664. GBS maintains GFPmut3 activity at different phases of growth. The application of fluorescence polarization enables easy discrimination of GBS GFPmut3 activity from the autofluorescence of culture media commonly used to grow GBS. Differential interference contrast microscopy, in combination with epifluorescence microscopy to detect GFPmut3 in GBS, enabled visualization of bacterial attachment to live human epithelial cells in real time. Plasmid pGU2664 was also used to visualize phenotypic differences in the adherence of wild-type GBS and an isogenic gene-deficient mutant strain lacking CovR (the control of virulence regulator) in adhesion assays. The system for GFPmut3 expression in GBS described in this study provides a new tool for the visualization of this organism in diverse research applications. We discuss the advantages and consider the limitations of this fluorescent biomarker system developed for GBS. IMPORTANCE Group B streptococcus (GBS) is a bacterium associated with various diseases in humans and animals. This study describes the development of a strategy to label and visualize GBS using a fluorescence biomarker, termed GFPmut3. We show that this biomarker can be successfully applied to track the growth of bacteria in liquid medium, and it enables the detailed visualization of GBS in the context of live human cells in real-time microscopic analysis. The system for GFPmut3 expression in GBS described in this study provides a new tool for the visualization of this organism in diverse research applications.
Hypertension Among Youths — United States, 2001–2016

Hypertension is an important modifiable risk factor for cardiovascular morbidity and mortality, and hypertension in adolescents and young adults is associated with long-term negative health effects (1,2). * In 2017, the American Academy of Pediatrics (AAP) released a new Clinical Practice Guideline (3), which updated 2004 pediatric hypertension guidance† with new thresholds and percentile references calculated from a healthy-weight population. To examine trends in youth hypertension and the impact of the new guideline on classification of hypertension status, CDC analyzed data from 12,004 participants aged 12-19 years in the 2001-2016 National Health and Nutrition Examination Survey (NHANES). During this time, prevalence of hypertension declined, using both the new (from 7.7% to 4.2%, p<0.001) and former (from 3.2% to 1.5%, p<0.001) guidelines, and declines were observed across all weight status categories. However, because of the new percentile tables and lower threshold for hypertension (4), application of the new guideline compared with the former guideline resulted in a weighted net estimated increase of 795,000 U.S. youths being reclassified as having hypertension using 2013-2016 data. Youths who were older, male, and those with obesity accounted for a disproportionate share of persons reclassified as having hypertension. Clinicians and public health professionals might expect to see a higher prevalence of hypertension with application of the new guideline and can use these data to inform actions to address hypertension among youths. Strategies to improve cardiovascular health include adoption of healthy eating patterns and increased physical activity (3).
CORL Expression and Function in Insulin Producing Neurons Reversibly Influences Adult Longevity in Drosophila

CORL proteins (known as SKOR in mice, Fussel in humans and fussel in Flybase) are a family of CNS specific proteins related to Sno/Ski oncogenes. Their developmental and adult roles are largely unknown. A Drosophila CORL (dCORL) reporter gene is expressed in all Drosophila insulin-like peptide 2 (dILP2) neurons of the pars intercerebralis (PI) of the larval and adult brain. The transcription factor Drifter is also expressed in the PI in a subset of dCORL and dILP2 expressing neurons and in several non-dILP2 neurons. dCORL mutant virgin adult brains are missing all dILP2 neurons that do not also express Drifter. This phenotype is also seen when expressing dCORL-RNAi in neurosecretory cells of the PI. dCORL mutant virgin adults of both sexes have a significantly shorter lifespan than their parental strain. This longevity defect is completely reversed by mating (lifespan increases over 50% for males and females). Analyses of dCORL mutant mated adult brains revealed a complete rescue of dILP2 neurons without Drifter. Taken together, the data suggest that dCORL participates in a neural network connecting the insulin signaling pathway, longevity and mating. The conserved sequence and CNS specificity of all CORL proteins imply that this network may be operating in mammals.
A blockchain-based PKI management framework

Public-Key Infrastructure (PKI) is the cornerstone technology that facilitates secure information exchange over the Internet. However, PKI is exposed to risks due to potential failures of Certificate Authorities (CAs) that may be used to issue unauthorized certificates for end-users. Many recent breaches show that if a CA is compromised, the security of the corresponding end-users will be in risk. As an emerging solution, Blockchain technology potentially resolves the problems of traditional PKI systems - in particular, elimination of single point-of-failure and rapid reaction to CAs shortcomings. Blockchain has the ability to store and manage digital certificates within a public and immutable ledger, resulting in a fully traceable history log. In this paper we designed and developed a blockchain-based PKI management framework for issuing, validating and revoking X.509 certificates. Evaluation and experimental results confirm that the proposed framework provides more reliable and robust PKI systems with modest maintenance costs.
Mapping Forest Structure Using UAS inside Flight Capabilities

We evaluated two unmanned aerial systems (UASs), namely the DJI Phantom 4 Pro and DJI Mavic Pro, for 3D forest structure mapping of the forest stand interior with the use of close-range photogrammetry techniques. Assisted flights were performed within two research plots established in mature pure Norway spruce (Picea abies (L.) H. Karst. ) and European beech (Fagus sylvatica L.) forest stands. Geotagged images were used to produce georeferenced 3D point clouds representing tree stem surfaces. With a flight height of 8 m above the ground, the stems were precisely modeled up to a height of 10 m, which represents a considerably larger portion of the stem when compared with terrestrial close-range photogrammetry. Accuracy of the point clouds was evaluated by comparing field-measured tree diameters at breast height (DBH) with diameter estimates derived from the point cloud using four different fitting methods, including the bounding circle, convex hull, least squares circle, and least squares ellipse methods. The accuracy of DBH estimation varied with the UAS model and the diameter fitting method utilized. With the Phantom 4 Pro and the least squares ellipse method to estimate diameter, the mean error of diameter estimates was −1.17 cm (−3.14%) and 0.27 cm (0.69%) for spruce and beech stands, respectively.
Association of HLA-DRB1 and -DQB1 alleles with type 1 (autoimmune) diabetes in African Arabs: systematic review and meta-analysis

ABSTRACT Several studies confirmed the association of HLA-DRB1 and -DQB1 alleles with altered risk of type 1 diabetes (T1D). However, data from individual studies based on small sample sizes yielded often conflicting findings in African Arabs. This is a systematic review and meta-analysis aimed at comprehensively evaluating this association with T1D, using molecular HLA data. Relevant studies were identified through systemic search of Medline/PubMed, Cochrane, Science Direct, ResearchGate, and EMBASE databases. Statistical analysis was carried out using RevMan, and Comprehensive Meta-analysis programs. Given the heterogeneity of African Arabs, we also performed subgroup analysis according to ethnicity. Analysis of sensitivity, heterogeneity, and publication bias were performed to validate the outcome of the findings. This meta-analysis included 862 T1DM cases, along with 1,390 normoglycemic control, and comprised ten comparisons. Our study indicates that DRB1*03 (OR = 2.86), DRB1*04 (OR = 2.78), and DQB1*02 (OR = 2.29), are positively associated with increased risk of T1DM, while DRB1*07 (OR = 0.48), DRB1*11 (OR = 0.20), DRB1*13 (OR = 0.47), DRB1*15 (OR = 0.30), DQB1*05 (OR = 0.39), and DQB1*06 (OR = 0.27) were negatively associated with T1D, suggesting a protective role against T1D. This meta-analysis was characterized by low heterogeneity, sensitivity, and publication bias, indicating the robustness and reliability of the results. Background: Several studies confirmed the association of HLA-DRB1 and –DQB1 alleles with altered risk of type 1 diabetes (T1D). However, data from individual studies based on small sample sizes yielded often conflicting findings in African Arabs. This is a systematic review and meta-analysis aimed at comprehensively evaluating this association with T1D, using molecular HLA data. Methods: Relevant studies were identified through systemic search of Medline/PubMed, Cochrane, Science Direct, ResearchGate, and EMBASE databases. Statistical analysis was carried out using Revman, and Comprehensive Meta-analysis programs. Given the heterogeneity of African Arabs, we also performed subgroup analysis according to ethnicity. Analysis of sensitivity, heterogeneity, and pub¬lication bias were performed to validate the outcome of the findings. This meta-analysis included 862 T1DM cases, along with 1,390 normoglycemic control, and comprised ten comparisons. Results: Our study indicates that DRB1*03 (OR = 2.86), DRB1*04 (OR = 2.78), and DQB1*02 (OR = 2.29), are positively associated with increased risk of T1DM, while DRB1*07 (OR = 0.48), DRB1*11 (OR = 0.20), DRB1*13 (OR = 0.47), DRB1*15 (OR = 0.30), DQB1*05 (OR = 0.39), and DQB1*06 (OR = 0.27) were negatively associated with T1D, suggesting a protective role against T1D. Conclusion: This meta-analysis was characterized by low heterogeneity, sensitivity, and publication bias, indicating the robustness and reliability of the results.
A Generic Mechanism for Perceptual Organization in the Parietal Cortex

Our visual system's ability to group visual elements into meaningful entities and to separate them from others is referred to as scene segmentation. Visual motion often provides a powerful cue for this process as parallax or coherence can inform the visual system about scene or object structure. Here we tested the hypothesis that scene segmentation by motion cues relies on a common neural substrate in the parietal cortex. We used fMRI and a set of three entirely distinct motion stimuli to examine scene segmentation in the human brain. The stimuli covered a wide range of high-level processes, including perceptual grouping, transparent motion, and depth perception. All stimuli were perceptually bistable such that percepts alternated every few seconds while the physical stimulation remained constant. The perceptual states were asymmetric, in that one reflected the default (nonsegmented) interpretation, and the other the non-default (segmented) interpretation. We confirmed behaviorally that upon stimulus presentation, the default percept was always perceived first, before perceptual alternations ensued. Imaging results showed that across all stimulus classes perceptual scene-segmentation was associated with an increase of activity in the posterior parietal cortex together with a decrease of neural signal in the early visual cortex. This pattern of activation is compatible with predictive coding models of visual perception, and suggests that parietal cortex hosts a generic mechanism for scene segmentation. SIGNIFICANCE STATEMENT Making sense of cluttered visual scenes is crucial for everyday perception. An important cue to scene segmentation is visual motion: slight movements of scene elements give away which elements belong to the foreground or background or to the same object. We used three distinct stimuli that engage visual scene segmentation mechanisms based on motion. They involved perceptual grouping, transparent motion, and depth perception. Brain activity associated with all three mechanisms converged in the same parietal region with concurrent deactivation of early visual areas. The results suggest that posterior parietal cortex is a hub involved in structuring visual scenes based on different motion cues, and that feedback modulates early cortical processing in accord with predictive coding theory.
Interaction of the α7-nicotinic subunit with its human-specific duplicated dupα7 isoform in mammalian cells: Relevance in human inflammatory responses

The α7 nicotinic receptor subunit and its partially duplicated human-specific dupα7 isoform are coexpressed in neuronal and non-neuronal cells. In these cells, α7 subunits form homopentameric α7 nicotinic acetylcholine receptors (α7-nAChRs) implicated in numerous pathologies. In immune cells, α7-nAChRs are essential for vagal control of inflammatory response in sepsis. Recent studies show that the dupα7 subunit is a dominant-negative regulator of α7-nAChR activity in Xenopus oocytes. However, its biological significance in mammalian cells, particularly immune cells, remains unexplored, as the duplicated form is indistinguishable from the original subunit in standard tests. Here, using immunocytochemistry, confocal microscopy, coimmunoprecipitation, FRET, flow cytometry, and ELISA, we addressed this challenge in GH4C1 rat pituitary cells and RAW264.7 murine macrophages transfected with epitope- and fluorescent protein-tagged α7 or dupα7. We used quantitative RT-PCR of dupα7 gene expression levels in peripheral blood mononuclear cells (PBMCs) from patients with sepsis to analyze its relationship with PBMC α7 mRNA levels and with serum concentrations of inflammatory markers. We found that a physical interaction between dupα7 and α7 subunits in both cell lines generates heteromeric nAChRs that remain mainly trapped in the endoplasmic reticulum. The dupα7 sequestration of α7 subunits reduced membrane expression of functional α7-nAChRs, attenuating their anti-inflammatory capacity in lipopolysaccharide-stimulated macrophages. Moreover, the PBMC's dupα7 levels correlated inversely with their α7 levels and directly with the magnitude of the patients' inflammatory state. These results indicate that dupα7 probably reduces human vagal anti-inflammatory responses and suggest its involvement in other α7-nAChR–mediated pathophysiological processes.
Automated In Vivo Sub-Hertz Analysis of Viscoelasticity (SAVE) for Evaluation of Breast Lesions

We present an automated method for acquiring images and contrast parameters based on mechanical properties of breast lesions and surrounding tissue at load frequencies less than 1 Hz. The method called sub-Hertz analysis of viscoelasticity (SAVE) uses a compression device integrated with ultrasound imaging to perform in vivo ramp-and-hold uniaxial creep-like test on human breast in vivo. It models the internal deformations of tissues under constant surface stress as a linear viscoelastic response. We first discuss different aspects of our unique measurement approach and the expected variability of the viscoelastic parameters estimated based on a simplified one-dimensional reconstruction model. Finite-element numerical analysis is used to justify the advantages of using imaging contrast over viscoelasticity values. We then present the results of SAVE applied to a group of patients with breast masses undergoing biopsy.
Cryopreserved Human Intestinal Mucosal Epithelium: A Novel In Vitro Experimental System for the Evaluation of Enteric Drug Metabolism, Cytochrome P450 Induction, and Enterotoxicity

We report here a novel in vitro enteric experimental system, cryopreserved human intestinal mucosa (CHIM), for the evaluation of enteric drug metabolism, drug-drug interaction, drug toxicity, and pharmacology. CHIM was isolated from the small intestines of four human donors. The small intestines were first dissected into the duodenum, jejunum, and ileum, followed by collagenase digestion of the intestinal lumen. The isolated mucosa was gently homogenized to yield multiple cellular fragments, which were then cryopreserved in a programmable liquid cell freezer and stored in liquid nitrogen. After thawing and recovery, CHIM retained robust cytochrome P450 (P450) and non-P450 drug-metabolizing enzyme activities and demonstrated dose-dependent induction of transcription of CYP24A1 (approximately 300-fold) and CYP3A4 (approximately 3-fold) by vitamin D3 as well as induction of CYP3A4 (approximately 3-fold) by rifampin after 24 hours of treatment. Dose-dependent decreases in cell viability quantified by cellular ATP content were observed for naproxen and acetaminophen, with higher enterotoxicity observed for naproxen, consistent with that observed in humans in vivo. These results suggest that CHIM may be a useful in vitro experimental model for the evaluation of enteric drug properties, including drug metabolism, drug-drug interactions, and drug toxicity.
Fetal surgery for spina bifida aperta

Spina bifida aperta (SBA) is one of the most common congenital malformations. It can cause severe lifelong physical and neurodevelopmental disabilities. Experimental and clinical studies have shown that the neurological deficits associated with SBA are not simply caused by incomplete neurulation at the level of the lesion. Additional damage is caused by prolonged exposure of the spinal cord and nerves to the intrauterine environment and a suction gradient due to cerebrospinal fluid leakage, leading to progressive downward displacement of the hindbrain. This natural history can be reversed by prenatal repair. A randomised controlled trial demonstrated that mid-gestational maternal-fetal surgery for SBA decreases the need for ventriculoperitoneal shunting and hindbrain herniation at 12 months and improves neurological motor function at 30 months of age. This came at the price of maternal and fetal risks, the most relevant ones being increased prematurity and a persistent uterine corporeal scar. Recently minimally invasive fetal approaches have been introduced clinically yet they lack extensive experimental or clinical trials. We aim to provide clinicians with the essential information necessary to counsel SBA parents as the basis for considering referral of selected patients to expert fetal surgery centres. We review the reported clinical outcomes and discuss recent developments of potentially less invasive fetal SBA approaches.
Protein Biomarkers of Cardiovascular Disease and Mortality in the Community

Background The discovery of novel and highly predictive biomarkers of cardiovascular disease (CVD) has the potential to improve risk‐stratification methods and may be informative regarding biological pathways contributing to disease. Methods and Results We used a discovery proteomic platform that targeted high‐value proteins for CVD to ascertain 85 circulating protein biomarkers in 3523 Framingham Heart Study participants (mean age, 62 years; 53% women). Using multivariable‐adjusted Cox models to account for clinical variables, we found 8 biomarkers associated with incident atherosclerotic CVD, 18 with incident heart failure, 38 with all‐cause mortality, and 35 with CVD death (false discovery rate, q<0.05 for all; P‐value ranges, 9.8×10−34 to 3.6×10−2). Notably, a number of regulators of metabolic and adipocyte homeostasis were associated with cardiovascular events, including insulin‐like growth factor 1 (IGF1), insulin‐like growth factor binding protein 1 (IGFBP1), insulin‐like growth factor binding protein 2 (IGFBP2), leptin, and adipsin. In a multimarker approach that accounted for clinical factors, growth differentiation factor 15 (GDF15) was associated with all outcomes. In addition, N‐terminal pro‐b‐type natriuretic peptide, C‐reactive protein, and leptin were associated with incident heart failure, and C‐type lectin domain family 3 member B (CLEC3B; tetranectin), N‐terminal pro‐b‐type natriuretic peptide, arabinogalactan protein 1 (AGP1), soluble receptor for advanced glycation end products (sRAGE), peripheral myelin protein 2 (PMP2), uncarboxylated matrix Gla protein (UCMGP), kallikrein B1 (KLKB1), IGFBP2, IGF1, leptin receptor, and cystatin‐C were associated with all‐cause mortality in a multimarker model. Conclusions We identified numerous protein biomarkers that predicted cardiovascular outcomes and all‐cause mortality, including biomarkers representing regulators of metabolic homeostasis and inflammatory pathways. Further studies are needed to validate our findings and define clinical utility, with the ultimate goal of improving strategies for CVD prevention.
Cellulose Synthase Stoichiometry Varies among Species and Tissues

Cellulose, the most abundant biopolymer on earth, is an important structural component of the primary and secondary cell wall of plant cells. It is also found in animals (tunicates), oomycetes, and bacteria ([Kumar and Turner, 2015][1]). Besides providing support and rigidity in living organisms,