Miracle-dz/UniFAR-FTUs huggingface dataset:
Cleaved N-terminal histone tails distinguish between NADPH oxidase (NOX)-dependent and NOX-independent pathways of neutrophil extracellular trap formation

Objectives Neutrophil extracellular traps (NETs) act in various rheumatic diseases. Although NET formation was originally described as a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX)-dependent pathway, it appears that there are also NOX-independent pathways of NET release. Currently, no tools are available that can discriminate between both NET-forming pathways. We aimed to develop a serological method allowing the discrimination between NETs generated through NOX-dependent or NOX-independent pathways. Methods Histones from in vitro generated NOX-dependent and NOX-independent NETs were characterised with a panel of lupus-derived antibodies against N-terminal histone tails using immunofluorescence microscopy, western blot and ELISA. NETs in patients with NET-associated diseases, that is, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), psoriatic arthritis (PsA) and sepsis, were characterised in sandwich ELISAs employing antibodies against myeloperoxidase (MPO) and N-terminal histone tails as detecting and capturing antibodies, respectively. Functional responses of endothelial cells to NOX-dependent and NOX-independent NETs were assessed as well. Results Neutrophil elastase cleaves the N-terminal tails of core histones during NOX-dependent, but not during NOX-independent NET formation. Consequently, the detection of MPO–histone complexes with antibodies against N-terminal histone tails allows discrimination between NETs formed through a NOX-dependent or NOX-independent manner. Characterisation of in vivo circulating NETs revealed the presence of NOX-independent NETs in RA, SLE and sepsis, but NOX-dependent NETs in PsA. NOX-independent NETs displayed an increased capacity to activate endothelial cells when compared with NOX-dependent NETs. Conclusions These results indicate heterogeneity in NET-forming pathways in vivo and highlight the need for disease-specific strategies to prevent NET-mediated pathology.
Reasons for low adherence to diet-diaries issued to pediatric dental patients: a collective case study

Objective Dietary habits are an important etiological factor in the development of dental caries. Several tools, such as 24-h dietary recall (retrospective) and diet-diaries (prospective), have been recommended for dietary assessment in dental practice. Diet-diaries are commonly advocated as a tool for oral health education; however, low adherence is found to be a recognized downside of their use in dental settings, as well as nutritional research more widely. However, the reasons for poor adherence to diet-diaries remain unclear. This study aimed to explore the reasons for poor adherence to diet-diaries issued to children in a dental hospital setting. Methods A qualitative collective case study design was employed to explore the use of diet-diaries as a health education tool. Twenty-eight data sources across 11 appointments included: observation of dentist–patient interactions, semi-structured interviews with child–parent dyads and dentists, in addition to documentary analysis of returned diet-diaries (this included 11 observations of dentist–patient interactions, 14 interviews with the child/parent dyads and dentists, and documentary analysis of three completed diet-diaries). Data from these multiple sources were integrated in a thematic analysis to identify themes and sub-themes. Results Two overarching themes were identified: 1) the diet-diary is perceived as a test which carries a potential for embarrassment and blame, which in turn generates defensive behavior from parents; and 2) parents’ values, priorities, and circumstances affect the level of commitment to completing a diet-diary. Conclusion Low adherence to diet-diary completion in clinical dentistry results from interacting factors related to the diet-diary itself, the patient, and the clinician. This study identifies a need for a more appropriate tool for dietary assessment that is patient-centered and compatible with modern lifestyles.
Assessment of Key Health and Wellness Indicators Among North Carolina Emergency Medical Service Providers

Abstract Objective: The objective of this study was to characterize key health indicators in Emergency Medical Services (EMS) personnel and identify areas for intervention in order to ensure a strong and capable emergency health workforce. Methods: Participants were EMS personnel delivering patients to 4 regional tertiary care emergency departments within North Carolina (NC). After transferring patient care and agreeing to participate, height, weight, and blood pressure (BP) measurements were recorded and each participant completed a questionnaire regarding demographics, activity levels, alcohol consumption, smoking, and medical history. Data were analyzed descriptively. Results: A sample of 452 EMS personnel from across NC was enrolled. The cohort was predominantly male (74.1%) and employed full-time (85.5%). The prevalence of overweight and obesity (80.3%) among EMS personnel was higher than the NC population (65.6%) and the general United States (US) population (70.8%). A previous diagnosis of high BP was reported by only 18.3% of participants, but 65.1% had elevated BP at the time of measurement. Alcohol consumption in the past 30 days among participants (55.4%) was slightly higher than state estimates (48.0%) and similar to national estimates (57.1%). However, heavy drinking (22.2%) and binge drinking (28.8%) were reported at much higher rates than state (5.6% and 15.2%, respectively) and national (6.6% and 18.3%, respectively) estimates. The prevalence of current smoking (21.5%) and quit attempts (48.8%) in the cohort was similar to state (21.8% and 55.0%, respectively) and national (21.2% and 55.7%, respectively) estimates. Likewise, the proportion of EMS providers meeting the Center for Disease Control’s activity guidelines (49.6%) was similar to that found in the NC (46.8%) and the general US (48.0%) populations. Conclusions: These findings suggest a high prevalence of overweight and obesity, heavy drinking, binge drinking, and high BP among NC EMS personnel. Similar to fire service personnel, these rates are higher than the general US population. As such, they suggest areas where intervention would have the greatest positive impact on the health and performance of the EMS workforce.
Factors that Affect the Osteoclastogenesis of RAW264.7 Cells

Osteoclasts and their activity are key regulators of bone formation. However, studying osteoclasts is difficult. Primary osteoclast cultures are difficult to maintain and isolate. Also, the amount of cells that are isolated and their properties depend on the origin and differentiation protocols. These protocols are usually developed in a distinct lab and multiple protocols exist. A cell line to study osteoclasts and a thorough study of osteoclast differentiation and culturing is currently lacking. The RAW264.7 cell line is most commonly used to study osteoclast differentiation and its signaling pathways. RAW264.7 cells are not a homogenous cell line. They don’t often exclusively differentiate into osteoclast but also into other multinucleated cells as well including macrophage polykaryons. A challenge of culturing RAW264.7 cells are culture conditions. Different conditions can affect survival, proliferation, and differentiation of RAW264.7 cells. Currently published protocols of culturing RAW264.7 cells often assume multinucleated cells that have three or more nuclei with distinguished osteoclast characteristics (such as TRAP+) as osteoclasts. However, osteoclasts and macrophage polykaryons are almost indistinguishable under a light microscope (TRAP+ with three or more nuclei). The goal of this paper is to examine the effect of culture conditions on the osteoclastogenesis ability of RAW264.7 cells. The focus will be on establishing the crucial parameters for culture density, time of stimulation, RANKL, and L-Gln concentrations. Although we are unable to establish the condition that offers a homogenous population of osteoclasts; nevertheless, we are able to identify the optimal conditions at which osteoclasts are found to be more than macrophage polykaryons. Finally, this article also demonstrates that osteoclasts and macrophage polykaryons can be distinguished by immunofluorescence staining for cathepsin K.
Genetic diversity analyses of Lasiodiplodia theobromae on Morus alba and Agave sisalana based on RAPD and ISSR molecular markers

ABSTRACT Genetic diversity of 23 Lasiodiplodia theobromae isolates on Morus alba and 6 isolates on Agave sisalana in Guangxi province, China, was studied by using random amplified polymorphic DNA and inter-simple sequence repeat molecular markers. Results of two molecular markers showed that the average percentage of polymorphic loci of all isolates was more than 93%. Both dendrograms of two molecular markers showed obvious relationship between groups and the geographical locations where those strains were collected, among which, the 23 isolates on M. alba were divided into 4 populations and the 6 isolates on A. sisalana were separated as a independent population. The average genetic identity and genetic distance of 5 populations were 0.7215, 0.3284 and 0.7915, 0.2347, respectively, which indicated that the genetic identity was high and the genetic distance was short in the 5 populations. Average value of the gene diversity index (H) and the Shannon’s information index (I) of 29 isolates were significantly higher than 5 populations which showed that genetic diversity of those isolates was richer than the populations and the degree of genetic differentiation of the isolates was higher. The Gst and Nm of 29 isolates were 0.4411, 0.6335 and 0.4756, 0.5513, respectively, which showed that the genetic diversity was rich in those isolates.
Perturbations of BMP/TGF-β and VEGF/VEGFR signalling pathways in non-syndromic sporadic brain arteriovenous malformations (BAVM)

Background Brain arteriovenous malformations (BAVM) represent a congenital anomaly of the cerebral vessels with a prevalence of 10–18/100 000. BAVM is the leading aetiology of intracranial haemorrhage in children. Our objective was to identify gene variants potentially contributing to disease and to better define the molecular aetiology underlying non-syndromic sporadic BAVM. Methods We performed whole-exome trio sequencing of 100 unrelated families with a clinically uniform BAVM phenotype. Pathogenic variants were then studied in vivo using a transgenic zebrafish model. Results We identified four pathogenic heterozygous variants in four patients, including one in the established BAVM-related gene, ENG, and three damaging variants in novel candidate genes: PITPNM3, SARS and LEMD3, which we then functionally validated in zebrafish. In addition, eight likely pathogenic heterozygous variants (TIMP3, SCUBE2, MAP4K4, CDH2, IL17RD, PREX2, ZFYVE16 and EGFR) were identified in eight patients, and 16 patients carried one or more variants of uncertain significance. Potential oligogenic inheritance (MAP4K4 with ENG, RASA1 with TIMP3 and SCUBE2 with ENG) was identified in three patients. Regulation of sma- and mad-related proteins (SMADs) (involved in bone morphogenic protein (BMP)/transforming growth factor beta (TGF-β) signalling) and vascular endothelial growth factor (VEGF)/vascular endotheliual growth factor recepter 2 (VEGFR2) binding and activity (affecting the VEGF signalling pathway) were the most significantly affected biological process involved in the pathogenesis of BAVM. Conclusions Our study highlights the specific role of BMP/TGF-β and VEGF/VEGFR signalling in the aetiology of BAVM and the efficiency of intensive parallel sequencing in the challenging context of genetically heterogeneous paradigm.
Clinical outcomes after initial treatment of methicillin-resistant Staphylococcus aureus infections

Objective To evaluate the clinical outcomes associated with anti-methicillin-resistant Staphylococcus aureus (MRSA) antimicrobials. Methods We reviewed a prospective database of 247 consecutive patients with clinically and microbiologically confirmed MRSA infections, hospitalized in 7 Japanese hospitals between April 2014 and March 2015, and treated with anti-MRSA pharmaceuticals. Survival was measured at 30 days. We examined the relationships between initial antimicrobial administered and survival and organ toxicity. HR and 95% CIs were calculated. Results Overall 30-day mortality was 12%. The lungs were infected in 105 (41%), skin and soft tissue in 73 (30%), and bones and joints in 21 (9%) patients. Bacteremia complicated the illness in 69 patients (28%). Among 5 pharmaceuticals, vancomycin was prescribed to 174 (71%), linezolid to 38 (16%), teicoplanin to 22 (9%), and daptomycin to 11 (5%) patients. Vancomycin tended to be associated with the lowest survival (HR=2.47; 95% CI=0.93–6.51; P=0.067), particularly in the lung-infected subgroup (HR=4.85; 95% CI=1.12–20.94; P=0.034) after adjustments for baseline illness severity. The incidence of renal dysfunction tended to be higher in patients with trough serum concentrations of vancomycin >15 mg/dL. Conclusion In this observational study reflecting real-world conditions, vancomycin was associated with higher 30-day mortality and incidence of kidney dysfunction than other anti-MRSA agents. The significance of the differences observed among antimicrobials other than vancomycin is uncertain.
Seasonal Variation in Human Adenovirus Conjunctivitis: A 30-Year Observational Study

ABSTRACT Purpose: The aim of this study is to measure the seasonal trends of adenovirus conjunctivitis. Methods: A total of 926 positive cases of HAdV conjunctivitis were reviewed at the University of Pittsburgh Medical Center (UPMC) in Pittsburgh (Allegheny County, Pennsylvania, USA) from 11th March 1987 to 15th December 2016. To determine seasonal variation, daily counts were adjusted for yearly changes in Allegheny County population size, modelled as a function of long-term trend using Basis-spline (B-spline), and analysed for seasonal trend by quarter (1Q = January to March, 2Q = April to June, 3Q = July to September, 4Q = October to December). Results: The number of adenovirus conjunctivitis cases in 3Q was 45% higher than that of 2Q (95% confidence interval [CI] 1.21–1.74, P = 0.0001). When 3Q is compared to 1Q and 4Q, it was not statistically significant (95% CI 0.96–1.42, P = 0.1209 and 95% CI 0.96–1.41, P = 0.1180, respectively). Conclusion: To the best of our knowledge, this is the first report and the longest time-series evaluating the potential seasonality of adenovirus conjunctivitis in the USA. We observed the highest number of cases from July to September and the lowest number of cases from April to June in our study population. Knowledge of seasonality can be used to guide outbreak expectations, limit antibiotic over-prescription, and enhance disease prevention.
Magnetic resonance T1-mapping evaluates the degree of thyroid destruction in patients with autoimmune thyroiditis

Objective Autoimmune thyroiditis (AIT) is the most common autoimmune thyroid disease. Longitudinal relaxation time mapping (T1-mapping) measured by MRI is a new technique for assessing interstitial fibrosis of some organs, such as heart and liver. This study aimed to evaluate the relationship between T1-mapping value and thyroid function and determine the usefulness of T1-mapping in identifying thyroid destruction in AIT patients. Methods This case–control study recruited 57 drug-naïve AIT patients and 17 healthy controls. All participants were given thyroid MRI, and T1-mapping values were measured using a modified look-locker inversion-recovery sequence. Results AIT patients had significantly higher thyroid T1-mapping values than the healthy controls (1.077 ± 177 vs 778 ± 82.9 ms; P < 0.01). A significant increase in thyroid T1-mapping values was presented along with the increased severity of thyroid dysfunction (P < 0.01). Correlation analyses showed that increased thyroid T1-mapping values were associated with higher TSH and lower FT3 and FT4 levels (TSH: r = 0.75; FT3: r = −0.47; FT4: r = −0.72; all P < 0.01). Receiver-operating characteristic curve analysis revealed a high diagnostic value of T1-mapping values for the degree of thyroid destruction (area under the curve was 0.95, 95% CI: 0.90–0.99, P < 0.01). Conclusions AIT patients have higher thyroid T1-mapping values than the healthy controls, and the T1-mapping values increased with the progression of thyroid dysfunction. Thyroid T1-mapping value might be a new index to quantitatively evaluate the degree of thyroid destruction in AIT patients.
Oral administration of titanium dioxide nanoparticle through ovarian tissue alterations impairs mice embryonic development

Background: Titanium dioxide nanoparticle (TiO2NP) is commonly used in industrial products including food colorant, cosmetics, and drugs. Previous studies have shown that oral administration of TiO2NP can be toxic to the reproductive system, but little is known if TiO2NP could be able to affect the functions of the female reproductive system, in particular fertility. Objective: The objective was to evaluate the effects of oral administration of TiO2NP on histological changes in ovaries, pregnancy rate and in vitro fertility in mice. Materials and Methods: In this experimental study, 54 adult female NMRI mice were randomly assigned to two groups: control group (received vehicle orally) and TiO2NP group (received 100 mg/kg/daily TiO2NP solution orally). After 5 wk, pregnancy and in vitro fertilization rates, histological changes in ovaries, malondyaldehyde and estrogen hormone levels in the blood serum were investigated and compared between groups. Results: Our results revealed that TiO2NP administration induced histological alterations in ovary including, degenerating and reduction of ovarian follicles, ovarian cyst formation and disturbance of follicular development. Compared to control, animals in TiO2NP group have shown significant reduction of pregnancy rates and number of giving birth (p=0.04). TiO2NP caused significant reduction in oocyte number, fertilization rate, and pre-implantation embryo development (p<0.001). Furthermore, malondyaldehyde and estrogen hormone levels were significantly (p<0.01) increased in mice received TiO2NP. Conclusion: Our findings suggest that TiO2NP exposure induces alterations on mice ovary resulting in a decrease in the rate of embryo development and fertility.
Adhesion awareness in 2016: An update of the national survey of surgeons

Background and aims Adhesions, that form in 60–80% of all abdominal operations, can cause complications such as chronic abdominal pain, small-bowel obstruction, female infertility, and the need for adhesiolysis in future surgeries. Our 2010 Adhesion Awareness survey demonstrated that despite the huge clinical impact of adhesions; adhesion-related complications were seldom mentioned in the informed consent. Six years later, a follow-up survey was conducted to assess the progress on awareness on adhesion-related complications in the Netherlands. Material and methods The 2010 Adhesion Awareness survey was repeated after a literature update. The knowledge regarding adhesions; the use of anti-adhesive agents and involvement in the informed consent process were assessed. Surgeons and surgical trainees were contacted by e-mail. The data was analysed using a Chi-square or Mann-Whitney U test and corrected for multiple testing. Results The response rate was 32.6%, similar to the survey in 2010 (34.4%). 88.1% agreed with the clinical relevance of adhesions, comparable to 2010 (89.8%). The score on the knowledge test was 38.8% (2010: 37.2%). Involvement of adhesion-related complications in the informed consent process increased, although 32.5% almost never mentions adhesions. In 2016, 42.4% reported a correct occurrence of bowel lesions during adhesiolysis, higher than in 2010 (P<0.001). Conclusions The adhesion awareness did not increase in six years, despite the efforts made. However, an increased awareness regarding adhesiolysis related complications was detected. Improvement of knowledge and behavior is essential to narrowing the gap between the impact of adhesions as a major complication of abdominal surgery and the limited adhesion awareness.
Human cancer evolution in the context of a human immune system in mice

Immunotherapy is one of the most promising cancer treatment modalities, but the lack of appropriate preclinical in vivo models hampers the development of novel immunotherapeutic strategies. Here, we studied the ability of transplanted human cancer cells to form primary tumors and metastasize in humanized immune system (HIS) mice created by transfer of CD34+ human hematopoietic stem cells. All tested transplanted cancer cell lines developed primary tumors that progressed nearly synchronously. Spontaneous lung and liver metastases developed from both orthotopic and ectopic transplanted cancer cells, and the ability to spread inversely correlated with the extent of CD8+ infiltration in the primary tumor. Further analysis revealed that interactions between the cancer model and the tumor‐infiltrating lymphocytes created tumor microenvironments (TMEs) resembling clinical cancers. Some models were largely immune cell‐excluding, while others appeared to develop adaptive resistance to immune‐mediated destruction by increased expression of programmed death ligand 1 (PDL1) and recruitment of human regulatory T cells. Our data suggest that HIS mice may provide a promising in vivo tumor model for evaluating immune modulatory anticancer therapies. Moreover, our study identified different tumor models resembling specific types of human TMEs, rendering each beneficial for addressing disease‐specific issues.
Scalable Generation of Mesenchymal Stem Cells from Human Embryonic Stem Cells in 3D

Human embryonic stem cell (hESC) derived mesenchymal stem cells (EMSC) are efficacious in treating a series of autoimmune, inflammatory, and degenerative diseases in animal models. However, all the EMSC derivation methods reported so far rely on two-dimensional (2D) culture systems, which are inefficient, costive and difficult for large-scale production. HESC, as an unlimited source, can be successively propagated in spheroids. Here, we demonstrate that hESC spheroids can directly differentiate into MSC spheroids (EMSCSp) within 20 days in one vessel without passaging and the system is scalable to any desired size. EMSCSp can further differentiate into osteocytes and chondrocytes in spheres or demineralized bone matrix. EMSCSp also retains immune-modulatory effects in vitro and therapeutic effects on two mouse models of colitis after dissociation. Compared to EMSC differentiated in monolayer, EMSCSp-derived cells have faster proliferation and higher yield and develop less apoptosis and slower senescence. Thus, the 3D differentiation system allows simple, cost-effective, and scalable production of high-quality EMSC and subsequently bone and cartilage tissues for therapeutic application.
Age of Onset of Alcohol Consumption and Subsequent Negative Health Outcomes in Gay and Bisexual Men Who Have Sex With Men

ABSTRACT The correlation between current alcohol use and sexual risk behaviors among men who have sex with men (MSM) is well documented, but little is known about age of drinking onset and current risks among this population. Data from a 2008 Internet survey of 8,452 MSM in the United States were used to assess factors associated with age of onset of alcohol use. In a multivariable model, drinking onset before age 15 was significantly associated with a higher number of lifetime male anal sex partners and being behaviorally bisexual in the past 12 months. Men who reported depressive symptoms in the past two weeks or who screened positive for alcohol abuse in the past 60 days were more likely to have early age of drinking onset. Findings suggest the need for targeting younger adolescents for alcohol screening, particularly sexual minority youth, to prevent risk behaviors in adulthood.