Towards improving online learning in physical education: Gender differences and determinants of motivation, psychological needs satisfaction, and academic achievement in Saudi students.

Studies on students' perceptions and expectations during physical education (PE) online learning remain scarce. Centered on self-determination theory, the present cross-sectional study aims to identify gender differences and predictors affecting motivation, psychological needs satisfaction (PNS), and academic achievement during PE online learning. Data were collected from Saudi students' (N = 308, 161 females and 147 males) responses to the PE autonomy, relatedness, competence, and motivation questionnaires. Welch's t-test for unequal sample sizes, multiple linear regression, and binary logistic regression were used to compare means and to predict the relationships between the independent and dependent variables. The results showed higher autonomy and competence perceptions in female than in male students, but no differences were observed in relatedness. Female students presented higher intrinsic motivations, lower amotivation perceptions than males. However, no gender differences were recorded in extrinsic motivation. Students with less experience in online learning and weak grade point averages (GPAs) are more susceptible to having a high level of amotivation. Gender, GPA, and prior experience with online learning are the common predictors for all PNS and amotivation, while GPA and prior experience with online learning are the determinants of intrinsic motivation. GPA is affected by prior experience with online learning, autonomy, competence, intrinsic motivation, and amotivation. Therefore, teachers are encouraged to adapt their didactic-pedagogical behaviors during PE online learning according to students' motivation and autonomy perceptions. Structuring teaching activities with more individualized support for autonomy, competence, intrinsic motivation, and students' online skills/competencies ensures better learning efficiency and academic achievements.

Anti-persister efficacy of colistin and meropenem against uropathogenic Escherichia coli is dependent on environmental conditions.

Antibiotic persistence is a phenomenon observed when genetically susceptible cells survive long-term exposure to antibiotics. These 'persisters' are an intrinsic component of bacterial populations and stem from phenotypic heterogeneity. Persistence to antibiotics is a concern for public health globally, as it increases treatment duration and can contribute to treatment failure. Furthermore, there is a growing array of evidence that persistence is a 'stepping-stone' for the development of genetic antimicrobial resistance. Urinary tract infections (UTIs) are a major contributor to antibiotic consumption worldwide, and are known to be both persistent (i.e. affecting the host for a prolonged period) and recurring. Currently, in clinical settings, routine laboratory screening of pathogenic isolates does not determine the presence or the frequency of persister cells. Furthermore, the majority of research undertaken on antibiotic persistence has been done on lab-adapted bacterial strains. In the study presented here, we characterized antibiotic persisters in a panel of clinical uropathogenic Escherichia coli isolates collected from hospitals in the UK and Australia. We found that a urine-pH mimicking environment not only induces higher levels of antibiotic persistence to meropenem and colistin than standard laboratory growth conditions, but also results in rapid development of transient colistin resistance, regardless of the genetic resistance profile of the isolate. Furthermore, we provide evidence for the presence of multiple virulence factors involved in stress resistance and biofilm formation in the genomes of these isolates, whose activities have been previously shown to contribute to the formation of persister cells.

Serum metabolomic profiles and semaphorin-3A as biomarkers of diabetic retinopathy progression.

Diabetic retinopathy (DR) is a typical microvascular complication of diabetes mellitus (DM) and it remains one of the leading causes of vision loss worldwide. Studies postulated that a distinct metabolic signature of DR exists and can be resolved from that of diabetes alone. Serum Semaphorin3A (Sema3A) levels have also been found to be correlated with the phenotypes of diabetic retinopathy. We aimed to analyze and identify serum metabolites and serum Sema3A levels that could be useful biomarkers of DR progression. This cross-sectional study included 45 type 2 diabetes (T2D) patients. Diabetic patients were divided into three groups based on the status of their complications: non-DR (NDR, n=15), non-proliferative DR (NPDR, n=15), and proliferative DR (PDR, n=15) groups. Serum metabolomic profiles of these patients were determined by using high-performance liquid chromatography-mass spectrometry (HPLC-MS), and serum Sema3A levels measured by ELISA. Metabolomics analysis revealed a set of metabolites that were altered in the serum of PDR patients as compared with NPDR and NDR groups. Among these metabolites total asymmetric dimethylarginine (ADMA) and Kynurenine were potential predictors of PDR patients. Significantly higher serum levels of Sema3A in PDR patients as compared with NPDR and NDR groups (p < 0.001), their serum levels were positively correlated with the central macular thickness (r= 0.952, p < 0.001) and negatively correlated with the superficial macular density (r=-0.952, p < 0.001). In conclusion, the metabolite signatures identified in this study and serum Sema3A levels could be proposed as biomarkers for DR development and progression in T2D patients. However, Sema3A was superior to metabolomics in the prediction of the severity of DR.

Rapid culture-independent loop-mediated isothermal amplification detection of antimicrobial resistance markers from environmental water samples.

Environmental water is considered one of the main vehicles for the transmission of antimicrobial resistance (AMR), posing an increasing threat to humans and animals health. Continuous efforts are being made to eliminate AMR; however, the detection of AMR pathogens from water samples often requires at least one culture step, which is time-consuming and can limit sensitivity. In this study, we employed comparative genomics to identify the prevalence of AMR genes within among: Escherichia coli, Klebsiella, Salmonella enterica and Acinetobacter, using publicly available genomes. The mcr-1, blaKPC (KPC-1 to KPC-4 alleles), blaOXA-48, blaOXA-23 and blaVIM (VIM-1 and VIM-2 alleles) genes are of great medical and veterinary significance, thus were selected as targets for the development of isothermal loop-mediated amplification (LAMP) detection assays. We also developed a rapid and sensitive sample preparation method for an integrated culture-independent LAMP-based detection from water samples. The developed assays successfully detected the five AMR gene markers from pond water within 1 h and were 100% sensitive and specific with a detection limit of 0.0625 µg/mL and 10 cfu/mL for genomic DNA and spiked bacterial cells, respectively. The integrated detection can be easily implemented in resource-limited areas to enhance One Health AMR surveillances and improve diagnostics.

Portable Differential Detection of CTX-M ESBL Gene Variants, blaCTX-M-1 and blaCTX-M-15, from Escherichia coli Isolates and Animal Fecal Samples Using Loop-Primer Endonuclease Cleavage Loop-Mediated Isothermal Amplification.

Cefotaximase-Munich (CTX-M) extended-spectrum beta-lactamase (ESBL) enzymes produced by Enterobacteriaceae confer resistance to clinically relevant third-generation cephalosporins. CTX-M group 1 variants, CTX-M-1 and CTX-M-15, are the leading ESBL-producing Enterobacteriaceae associated with animal and human infection, respectively, and are an increasing antimicrobial resistance (AMR) global health concern. The blaCTX-M-1 and blaCTX-M-15 genes encoding these variants have an approximate nucleotide sequence similarity of 98.7%, making effective differential diagnostic monitoring difficult. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) enables rapid real-time multiplex pathogen detection with single-base specificity and portable on-site testing. We have developed an internally controlled multiplex CTX-M-1/15 LEC-LAMP assay for the differential detection of blaCTX-M-1 and blaCTX-M-15. Assay analytical specificity was established using a panel of human, animal, and environmental Escherichia coli isolates positive for blaCTX-M-1 (n = 18), blaCTX-M-15 (n = 35), and other closely related blaCTX-Ms (n = 38) from Ireland, Germany, and Portugal, with analytical sensitivity determined using probit regression analysis. Animal fecal sample testing using the CTX-M-1/15 LEC-LAMP assay in combination with a rapid DNA extraction protocol was carried out on porcine fecal samples previously confirmed to be PCR-positive for E. coli blaCTX-M. Portable instrumentation was used to further analyze each fecal sample and demonstrate the on-site testing capabilities of the LEC-LAMP assay with the rapid DNA extraction protocol. The CTX-M-1/15 LEC-LAMP assay demonstrated complete analytical specificity for the differential detection of both variants with sensitive low-level detection of 8.5 and 9.8 copies per reaction for blaCTX-M-1 and blaCTX-M-15, respectively, and E. coli blaCTX-M-1 was identified in all blaCTX-M positive porcine fecal samples tested. IMPORTANCE CTX-M ESBL-producing E. coli is an increasing AMR public health issue with the transmission between animals and humans via zoonotic pathogens now a major area of interest. Accurate and timely identification of ESBL-expressing E. coli CTX-M variants is essential for disease monitoring, targeted antibiotic treatment and infection control. This study details the first report of portable diagnostics technology for the rapid differential detection of CTX-M AMR markers blaCTX-M-1 and blaCTX-M-15, facilitating improved identification and surveillance of these closely related variants. Further application of this portable internally controlled multiplex CTX-M-1/15 LEC-LAMP assay will provide new information on the transmission and prevalence of these CTX-M ESBL alleles. Furthermore, this transferable diagnostic technology can be applied to other new and emerging relevant AMR markers of interest providing more efficient and specific portable pathogen detection for improved epidemiological surveillance.

ß-Lactam resistance genes present in UK pheasants and red-legged partridges.

BACKGROUND: Despite considerable recent reductions in antimicrobial use, the UK gamebird industry continues to struggle with production diseases during the rearing season, necessitating significant antibiotic use. This observational study investigated the presence of genes conferring resistance to ß-lactam antibiotics within industry-reared pheasants and red-legged partridges in the UK. METHODS: DNA was extracted from 60 pooled caecal samples collected from gamebirds at routine postmortem examinations during the rearing season. Genes encoding extended-spectrum ß-lactamases (ESBL) were detected by PCR and the corresponding alleles were determined. RESULTS: Over half (53%) of the samples harboured genes encoding blaTEM resistance, with blaSHV identified in 20% of samples. The blaTEM gene was more common on sites with higher antibiotic use, whereas blaSHV was predominantly found in birds younger than 5 weeks. Genotyping of the identified resistance genes revealed the presence of blaTEM-1 , blaSHV-1 and blaSHV-11 alleles. LIMITATIONS: This was a small-scale study conducted at four sites in southern England. CONCLUSION: This is the first report of the presence of ESBL genes in gamebirds, highlighting the need for further research into antimicrobial resistance in UK gamebirds.

Mapping the evidence of the effects of environmental factors on the prevalence of antibiotic resistance in the non-built environment: Protocol for a systematic evidence map.

BACKGROUND: Human, animal, and environmental health are increasingly threatened by the emergence and spread of antibiotic resistance. Inappropriate use of antibiotic treatments commonly contributes to this threat, but it is also becoming apparent that multiple, interconnected environmental factors can play a significant role. Thus, a One Health approach is required for a comprehensive understanding of the environmental dimensions of antibiotic resistance and inform science-based decisions and actions. The broad and multidisciplinary nature of the problem poses several open questions drawing upon a wide heterogeneous range of studies. OBJECTIVE: This study seeks to collect and catalogue the evidence of the potential effects of environmental factors on the abundance or detection of antibiotic resistance determinants in the outdoor environment, i.e., antibiotic resistant bacteria and mobile genetic elements carrying antibiotic resistance genes, and the effect on those caused by local environmental conditions of either natural or anthropogenic origin. METHODS: Here, we describe the protocol for a systematic evidence map to address this, which will be performed in adherence to best practice guidelines. We will search the literature from 1990 to present, using the following electronic databases: MEDLINE, Embase, and the Web of Science Core Collection as well as the grey literature. We shall include full-text, scientific articles published in English. Reviewers will work in pairs to screen title, abstract and keywords first and then full-text documents. Data extraction will adhere to a code book purposely designed. Risk of bias assessment will not be conducted as part of this SEM. We will combine tables, graphs, and other suitable visualisation techniques to compile a database i) of studies investigating the factors associated with the prevalence of antibiotic resistance in the environment and ii) map the distribution, network, cross-disciplinarity, impact and trends in the literature.

Prognostic Significance of Platelet Activation Marker CD62P in Hospitalized Covid-19 Patients.

BACKGROUND: CD62P is a surface marker for platelet activation. Platelet dysfunction contributes to disproportionate intravascular microthrombosis in SARS-CoV-2. We aimed to assess the clinical significance of CD62P as a biomarker of platelet activation and its correlation to the clinical severity and outcome of COVID-19 infections. METHODS: The study included 80 COVID-19 patients and, in addition, there were 20 age and gender-matched healthy controls. Laboratory measurements included CBC, serum ferritin, LDH, CRP, D-dimer and flow cytometric assessments of the platelet markers CD42b and CD62P. The primary study outcome was patients' survival at the end of study. RESULTS: Among the studied patients, 24 patients (30.0%) died by the end of the study. Survivors had significantly lower CD62P values when compared with non-survivors [median (IQR): 75.5 (73.0 - 91.0) versus 96.0 (93.5 - 97.8), p < 0.001]. Patients with severe disease had significantly higher levels of CD62P levels [median (IQR): 95.5 (92.0 - 97.8) versus 75.0 (72.0 - 76.8), p < 0.001]. Logistic regression analysis identified D-dimer levels [OR (95% CI): 0.14 (0.03 - 0.74) and CD62P levels: OR (95% CI): 0.4 (0.17 - 0.94)] as significant predictors of mortality in multivariate analysis. CONCLUSIONS: CD62P expression on admission may be a useful prognostic maker in hospitalized Covid-19 patients. Its expression is related to other markers of inflammation and coagulopathy.

Determination of Therapeutic and Safety Effects of Zygophyllum coccineum Extract in Induced Inflammation in Rats.

Background: Z. coccineum is a facultative plant with many medicinal applications. This study examined the anti-inflammatory activity of Zygophyllum coccineum (Z. coccineum) in an arthritis animal model. Materials and Methods: Seventy-Six Wistar Albino rats of either sex randomly divided into six groups (12/each). The inflammation model was done using Complete Freund's Adjuvant in albino rats. The anti-inflammatory activities of the extract were estimated at different dose levels (15.6, 31, and 60 mg/kg) as well as upon using methotrexate (MTX) as a standard drug (0.3 mg/kg). Paw volume and arthritis index scores have been tested in all examined animals' treatments. Histological examination of joints was also performed. Flow cytometric studies were done to isolated osteoclasts. Cytokines assay as well as biochemical testing was done in the examined samples. Results. In vitro studies reported an IC50 of 15.6 µg/ml for Z. coccineum extract in lipoxygenase inhibition assay (L.O.X.). Moreover, it could be noticed that isorhamnetin-3-O-glucoside, tribuloside, and 7-acetoxy-4-methyl coumarin were the most common compounds in Z. coccineum extract separated using L.C.-ESI-TOF-M.S. (liquid chromatography-electrospray ionization ion-trap time-of-flight mass spectrometry). Microscopic examinations of synovial tissue and hind limb muscles revealed the effect of different doses of Z. coccineum extract on restoring chondrocytes and muscles structures. Osteoclast size and apoptotic rate examinations revealed the protective effect of Z. coccineum extract on osteoclast. The results upon induction of animals and upon treatment using of MTX significantly increased apoptotic rate of osteoclast compared to control, while using of 15.6 µg/ml. for Z. coccineum extract lead to recover regular apoptotic rate demonstrating the protective effect of the extract. Z. coccineum extract regulated the secretion of proinflammatory and anti-inflammatory cytokines. Biochemical tests indicated the safety of Z. coccineum extract on kidney and liver functions. Conclusion. Z. coccineum extract has efficient and safe anti-inflammatory potential in an induced rat model.

Maternal serum perlecan and ischemia modified albumin levels as biomarkers of preeclampsia severity.

Preeclampsia (PE) is a pregnancy disorder characterized by hypertension and end-organ damage. Reliable biochemical markers for diagnosis and prediction of PE severity can improve maternal health, and several of these markers have been suggested till now. The goal of our study was to evaluate maternal serum levels of Perlecan and Ischemia modified albumin (IMA) in PE patients, and to investigate their relationship with the severity. This study included 45 pregnant women, who were divided into three groups: mild PE (n=15), severe PE (n=10), and normal pregnant females (n=20) as a control group. Maternal serum levels of Perlecan and IMA were determined by the enzyme linked immunosorbent assay (ELISA). Preeclamptic women with severe features have significantly higher serum Perlecan and IMA levels than women with mild PE and control (P0.001 for both). Serum levels of Perlecan and IMA were significantly increased in patients with mild PE as compared with control (P0.001 for both). Serum Perlecan levels were positively correlated with systolic blood pressure (SBP), diastolic blood pressure (DBP), ALT, AST, creatinine, urea, uric acid, and proteinuria, but negatively correlated with platelet count and fetal birth weight. Serum IMA level was positively correlated with SBP, DBP, but negatively correlated with Albumin, and fetal birth weight. The receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of Perlecan and IMA in the prediction of PE severity. Serum Perlecan had greater sensitivity and lower specificity for severe PE than for mild PE. Serum IMA had greater sensitivity and lower specificity for severe PE than for mild PE. In conclusion, maternal serum Perlecan and IMA levels were biomarkers for monitoring PE and the increase in serum Perlecan levels was in accordance with the severity of PE. Also, Perlecan was superior to IMA as a predictor for PE severity.

JMM profile: Loop-mediated isothermal amplification (LAMP): for the rapid detection of nucleic acid targets in resource-limited settings.

Loop-mediated isothermal amplification (LAMP) is a rapid alternative to PCR, in which the reaction occurs at one temperature and uses a polymerase with high displacement activity, e.g. Bacillus stearothermophilus DNA polymerase I (Bst) or homologues. Since the discovery of LAMP in 2000, several applications have been developed to employ this technique in the rapid detection of nucleic acid targets and enhance its performance. Improvements to the LAMP technique and a variety of innovative detection methods have led to its application for a wide range of targets in medical and veterinary microbiology, particularly in resource-poor settings. The key advantages of LAMP-based diagnostics include the ability to rapidly detect target nucleic acid sequences within 30 min and its ease of use, facilitating its application in field, bedside, pen-side, point-of-care and point-of-need diagnostic settings. LAMP can be a valuable tool to aid in the detection and management of disease outbreaks.

Diet and physical exercises for preschoolers with ADHD and their mothers: An intervention study.

BACKGROUND: Attention deficit hyperactivity disorder (ADHD) is a serious public health problem. Diet-focused approaches and physical exercise can be used to complement other ADHD management techniques. OBJECTIVE: To determine the prevalence of ADHD symptoms among preschoolers in nursery schools and to evaluate the educational interventions toward nutrition and physical exercise in mothers and their preschoolers with ADHD symptoms. RESEARCH METHODOLOGY: A two-phase sampling method was employed. First, a cross-sectional survey was conducted to determine the prevalence of ADHD symptoms in four nursery schools (400 preschoolers aged between 3 and 6 years). Second, an intervention study (a quasi-experimental research design with one group completing the pre-test and the post-test) was performed on 36 preschoolers having ADHD symptoms and their mothers by using the educational intervention for mothers and photos and games about nutrition and physical exercise for the preschoolers with ADHD; mothers of four children out of the 40 refused to participate in the study. Data were analyzed using SPSS version 20. The paired t-test was used to determine significant differences between the groups. Differences were considered significant at P < 0.05. RESULTS: Of the 400 preschoolers, 10% had high ADHD symptoms. The mean score of mothers' knowledge of nutrition and physical exercise improved after the implementation of the program (p = 0.01). In addition, preschoolers with ADHD enjoyed the session with photos and games (p = 0.01). CONCLUSIONS AND IMPLICATIONS: Educational intervention significantly improved the knowledge of the mothers. Moreover, preschoolers with ADHD symptoms enjoyed the session with photos and games. This intervention appears to be feasible and promising for further investigation of its effects.

Study of transforming growth factor beta 1 gene +869T/C polymorphism in Egyptian rheumatoid arthritis patients and its relation to disease activity.

This study intended to explore the relationship between the +869T/C polymorphism of the transforming growth factor-ß1 (TGF-ß1) gene and rheumatoid arthritis (RA) predisposition and activity in Egyptian patients. The study involved 30 patients suffering from RA and 30 apparently healthy participants as the control group. The +869T/C polymorphism of the TGF-ß1 gene was determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) process. The TGF-ß1 + 869 CT genotype and CT+TT genotypes in RA patients showed a significant increase than the control group (OR=3.782 and 3.824, CI=1.046-13.680 and 1.150-12.713, P=0.043 and 0.029, respectively). T allele showed a significant increase in patients than in controls (OR= 2.104, CI 1.015- 4.361, P = 0.046). The TGF-ß1 +869 CT+TT genotypes were accompanied by higher DAS-28 scores which express higher disease activity, and increased levels of RF, Anti-CCP, ESR, and CRP. In conclusion, the TGF-ß1 +869T/C gene polymorphism may be accompanied by an increased predisposition to RA and with its severity in Egyptian RA patients.

Median Nerve Affection in Hypertensive Patients with and without Diabetes High-Resolution Ultrasound Assessment.

PURPOSE: Diabetes is a documented risk factor for peripheral neuropathy. It was reported that associated hypertension could increase this risk. The present study aimed to assess the effect of hypertension and diabetes on median nerve using high-resolution ultrasound. METHODS: The study includes 50 hypertensive patients (HTN group), 50 diabetic patients (DM group), 50 patients with coexisting diabetes and hypertension (HTN + DM group) and 50 healthy controls. Median nerve affection in the studied groups was studied by vibration perception thresholds (VPT). The median nerve cross-sectional area was determined at the nerve cross-sectional area of the median nerve at the carpal tunnel by high-resolution ultrasound. Clinical symptoms were assessed using Toronto Clinical Severity Score (TCSS). RESULTS: There was significantly higher median nerve CSA in all patient groups in comparison to controls. HTN + DM group had significantly higher median nerve CSA when compared with DM group. Patients with peripheral neuropathy in HTN + DM and DM groups had significantly higher median nerve CSA than patients without. Using ROC curve analysis, it was shown that median CSA could successfully distinguish patients with peripheral neuropathy from patients without in HTN + DM group [AUC (95% CI): 0.71 (0.54-0.89)] and in DM group [AUC (95% CI): 0.86 (0.72-0.99)]. CONCLUSION: Hypertensive patients with and without diabetes have significantly higher median nerve CSA when compared with controls.

An ex vivo cystic fibrosis model recapitulates key clinical aspects of chronic Staphylococcus aureus infection.

Staphylococcus aureus is the most prevalent organism isolated from the airways of people with cystic fibrosis (CF), predominantly early in life. Yet its role in the pathology of lung disease is poorly understood. In mice, and many experiments using cell lines, the bacterium invades cells or interstitium, and forms abscesses. This is at odds with the limited available clinical data: interstitial bacteria are rare in CF biopsies and abscesses are highly unusual. Bacteria instead appear to localize in mucus plugs in the lumens of bronchioles. We show that, in an established ex vivo model of CF infection comprising porcine bronchiolar tissue and synthetic mucus, S. aureus demonstrates clinically significant characteristics including colonization of the airway lumen, with preferential localization as multicellular aggregates in mucus, initiation of a small colony variant phenotype and increased antibiotic tolerance of tissue-associated aggregates. Tissue invasion and abscesses were not observed. Our results may inform ongoing debates relating to clinical responses to S. aureus in people with CF.

Predicting Antibiotic-Associated Virulence of Pseudomonas aeruginosa Using an ex vivo Lung Biofilm Model.

BACKGROUND: Bacterial biofilms are known to have high antibiotic tolerance which directly affects clearance of bacterial infections in people with cystic fibrosis (CF). Current antibiotic susceptibility testing methods are either based on planktonic cells or do not reflect the complexity of biofilms in vivo. Consequently, inaccurate diagnostics affect treatment choice, preventing bacterial clearance and potentially selecting for antibiotic resistance. This leads to prolonged, ineffective treatment. METHODS: In this study, we use an ex vivo lung biofilm model to study antibiotic tolerance and virulence of Pseudomonas aeruginosa. Sections of pig bronchiole were dissected, prepared and infected with clinical isolates of P. aeruginosa and incubated in artificial sputum media to form biofilms, as previously described. Then, lung-associated biofilms were challenged with antibiotics, at therapeutically relevant concentrations, before their bacterial load and virulence were quantified and detected, respectively. RESULTS: The results demonstrated minimal effect on the bacterial load with therapeutically relevant concentrations of ciprofloxacin and meropenem, with the latter causing an increased production of proteases and pyocyanin. A combination of meropenem and tobramycin did not show any additional decrease in bacterial load but demonstrated a slight decrease in total proteases and pyocyanin production. CONCLUSION: In this initial study of six clinical isolates of P. aeruginosa showed high levels of antibiotic tolerance, with minimal effect on bacterial load and increased proteases production, which could negatively affect lung function. Thus, the ex vivo lung model has the potential to be effectively used in larger studies of antibiotic tolerance in in vivo-like biofilms, and show how sub optimal antibiotic treatment of biofilms may potentially contribute to exacerbations and eventual lung failure. We demonstrate a realistic model for understanding antibiotic resistance and tolerance in biofilms clinically and for molecules screening in anti-biofilm drug development.

Urinary Orosomucoid - 2 and Soluble CD14 as Potential Biomarkers for Assessment of Disease Activity in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is the most common inflammatory joint disease leading to severe disability and premature mortality. Current blood biomarkers for assessing RA activity are invasive and are not highly sensitive or specific to changes in disease activity. Therefore, there is a need for new biomarkers that can accurately indicate disease activity. The present study evaluated the use of urinary orosomucoid (ORM) - 2 and soluble CD14 (sCD14) as non-invasive biomarkers for precise assessment of disease activity of RA, in order to improve treatment outcomes in RA patients. The study included 36 female patients with RA, were divided into three groups of mild, moderate and severe disease activity according to disease activity score 28 (DAS 28) based on erythrocyte sedimentation rate (ESR) and were compared to control group. Urinary levels of ORM-2 and sCD14 were measured by ELISA. All patients showed significant increase in urinary ORM-2 and sCD14 levels in comparison to controls. There were significant positive correlations of urinary ORM-2 and sCD14 levels with DAS28score and also with the conventional blood biomarkers (C- reactive protein (CRP) and ESR). The receiver operating characteristic curve analysis revealed that both urinary ORM-2 and sCD14 have higher predictive value for disease activity than CRP and ESR. In conclusion, Urinary ORM-2 and sCD14 levels were increased in patients with RA and were correlated with the disease activity. Thus, the urinary biomarkers might be able to replace blood measures for RA activity as they could provide non-invasive and precise assessment of disease activity in RA patients.

A nanoparticle-based method for culture-free bacterial DNA enrichment from whole blood.

Point-of-care (POC) diagnostics are one of the quick and sensitive detection approaches used in current clinical applications, but always face a performance tradeoff between time-to-result and assay sensitivity. One critical setting where these limitations are evident is the detection of sepsis, where 6-10mL of whole blood may contain as little as one bacterial colony forming unit (cfu). The large sample volume, complex nature of the sample and low analyte concentration necessitates signal enhancement using culture-based or molecular amplification techniques. In the time-critical diagnosis of sepsis, waiting for up to 24h to produce sufficient DNA for analysis is not possible. As a consequence, there is a need for integrated sample preparation methods that could enable shorter detection times, whilst maintaining high analytical performance. We report the development of a culture-free bacterial enrichment method to concentrate bacteria from whole blood in less than 3h. The method relies on triple-enrichment steps to magnetically concentrate bacterial cells and their DNA with a 500-fold reduction in sample volume (from 10 to 0.02mL). Using this sample preparation method, sensitive qPCR detection of the extracted S. aureus bacterial DNA was achieved with a detection limit of 5±0.58cfu/mL within a total elapsed time of 4h; much faster than conventional culture-based approaches. The method could be fully automated for integration into clinical practice for point-of-care or molecular detection of bacterial DNA from whole blood.

Surface Ligand Density of Antibiotic-Nanoparticle Conjugates Enhances Target Avidity and Membrane Permeabilization of Vancomycin-Resistant Bacteria.

Many bacterial pathogens have now acquired resistance toward commonly used antibiotics, such as the glycopeptide antibiotic vancomycin. In this study, we show that immobilization of vancomycin onto a nanometer-scale solid surface with controlled local density can potentiate antibiotic action and increase target affinity of the drug. Magnetic nanoparticles were conjugated with vancomycin and used as a model system to investigate the relationship between surface density and drug potency. We showed remarkable improvement in minimum inhibitory concentration against vancomycin-resistant strains with values of 13-28 µg/mL for conjugated vancomycin compared to 250-4000 µg/mL for unconjugated vancomycin. Higher surface densities resulted in enhanced affinity toward the bacterial target compared to that of unconjugated vancomycin, as measured by a competition experiment using a surrogate ligand for bacterial Lipid II, N-Acetyl-l-Lys-d-Ala-d-Ala. High density vancomycin nanoparticles required >64 times molar excess of ligand (relative to the vancomycin surface density) to abrogate antibacterial activity compared to only 2 molar excess for unconjugated vancomycin. Further, the drug-nanoparticle conjugates caused rapid permeabilization of the bacterial cell wall within 2 h, whereas no effect was seen with unconjugated vancomycin, suggesting additional modes of action for the nanoparticle-conjugated drug. Hence, immobilization of readily available antibiotics on nanocarriers may present a general strategy for repotentiating drugs that act on bacterial membranes or membrane-bound targets but have lost effectiveness against resistant bacterial strains.

Median nerve motor entry points in the forearm - clinical application

Superficial forearm flexors receive primary and secondary branches from the median nerve (mn). These branches vary in number, size and motor entry points (m) into the muscles. Knowledge of these points is essential for maximal compound muscle action potential (cmap) recording from these muscles. Spasticity of these flexors is treated using botulinum toxin (bt) injection or selective partial neurotomy (spn) of the nerve branches to the spastic muscles. Twenty human cadaveric forearms were dissected. The location of the motor entry points of the median nerve to the superficial forearm flexor muscles was expressed as a distance from the medial (me) and lateral (le) epicondyles of the humerus. Fifty apparently healthy volunteers (25 males and 25 females) underwent cmap recording from the superficial forearm flexors. Thirty patients (15 males and 15 females) with spastic hyperflexion of the wrist and fingers underwent bt injection or spn. Pronator teres (pt) had 2-4 m, flexor carpi radialis (fcr) had 1-3 m and flexor digitorum superficialis (fds) had 3-8 m. Variable shapes of the cmap were recorded from them (monophasic, biphasic or multiphasic). Based on the anatomical results, bt injection was done at 5 points (p1-p5); pt was injected at p1, fcr was injected at p2, and fds is a large muscle and was injected at p3, p4, p5 (proximal, middle, distal), giving good results in 85% of cases; spn was done in severe cases refractory to bt injection with excellent to good results in 80% of cases. The patterns of branching of mn differ from the classically described patterns. Therefore, revising the innervation patterns of the superficial forearm flexors is mandatory, since the variations observed are more diverse than has been described. Identification of the branches and the motor entry points of mn are essential for cmap recording from the superficial forearm flexors, bt injection, spn and tendon transfer

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