Miracle-dz/UniFAR-FTUs huggingface dataset:
Gap junction signalling mediated through connexin-43 is required for chick limb development.

During chick limb development the gap junction protein Connexin-43 (Cx43) is expressed in discrete spatially restricted domains in the apical ectodermal ridge (AER) and mesenchyme of the zone of polarising activity. Antisense oligonucleotides (ODNs) were used to investigate the role of Connexin-43 (Cx43) in the development of the chick limb bud. We have used unmodified ODNs in Pluronic F-127 gel, which is liquid at low temperature but sets at room temperature and so remains situated at the point of application. As a mild surfactant, the gel increases antisense ODN penetration and supplies ODNs to the embryo continually for 12-18 h. We have shown a strong decrease in Cx43 protein expression after application of specific antisense oligonucleotides but the abundance of a closely related protein, Connexin-32 (Cx32), was not affected. Application of antisense Cx43 ODNs at stages 8-15 HH before limb outgrowth resulted in dramatic limb phenotypes. About 40% of treated embryos exhibited defects such as truncation of the limb bud, fragmentation into two or more domains, or complete splitting of the limb bud into two or three branches. Molecular analysis of antisense treated embryos failed to detect Shh or Bmp-2 in anterior structures and suggested that extra lobes seen in nicked and split limbs were not a result of establishment of new signalling centres as found after the application of FGF to the flank. However, examination of markers for the AER showed a number of abnormalities. In severely truncated specimens we were unable to detect the expression of either Fgf-4 or Fgf-8. In both nicked and split limbs the expression of these genes was discontinuous. Down-regulation of Cx43 after the antisense application could be comparable to AER removal and results in distal truncation of the limb bud. Taken together these data suggest the existence of a feedback loop between the FGFs and signalling mediated by Cx43.
Serotype Identification of Group B Streptococci by PCR and Sequencing

ABSTRACT Group B streptococcus (GBS; Streptococcus agalactiae) is the most common cause of neonatal and obstetric sepsis and is an increasingly important cause of septicemia in elderly individuals and immunocompromised patients. Ongoing surveillance to monitor GBS serotype distribution will be needed to guide the development and use of GBS conjugate vaccines. We designed sequencing primers based on the previously published sequences of the capsular polysaccharide (cps) gene clusters to further define partial cps gene clusters for eight of the nine GBS serotypes (serotypes Ia to VII). Subsequently, we designed and evaluated primers to identify serotypes Ia, Ib, III, IV, V, and VI directly by PCR and all eight serotypes (serotypes Ia to VII) by sequence heterogeneity. A total of 206 clinical GBS isolates were used to compare our molecular serotype (MS) identification method with conventional serotyping (CS). All clinical isolates were assigned an MS, whereas 188 of 206 (91.3%) were assigned a serotype by use of antisera. A small number of isolates (serosubtypes III-3 and III-4) showed different serotype specificities between PCR and sequencing, but the PCR results correlated with those obtained by CS. The overall agreement between the MS identification method and CS for isolates for which results of both tests were available was 100% (188 of 188 isolates). The MS identification method is a specific and practical alternative to conventional GBS serotyping and will facilitate epidemiological studies.
Virion assembly defect of polyomavirus hr-t mutants: underphosphorylation of major capsid protein VP1 before viral DNA encapsidation

The major capsid protein of polyomavirus, VP1, was separated into at least four subspecies by isoelectric focusing. One of these subspecies was selectively extracted from purified virions by mild treatment with sodium dodecyl sulfate, leaving a 140S particle enriched in the other three forms. The two most acidic subspecies were labeled in vivo with [32P]phosphate, and these subspecies are among those identified as being deficient in nontransforming host range (hr-t) mutant virus nonpermissive infection of NIH3T3 cells. Quantitation of VP1 phosphorylation revealed that hr-t mutant virus VP1 is phosphorylated to about 40 to 50% the level of the wild type in NIH3T3 cells, and two-dimensional phosphoamino acid analysis suggested that threonine phosphorylation was affected more than serine phosphorylation. Two results indicate that the VP1 modifications occur before and independent of virus assembly: modified subspecies were detected during wild-type infection within a 2-min pulse-label with [32S]methionine, and VP1 modifications of temperature-sensitive VP1 mutants were the same at both restrictive and permissive temperatures for virus assembly. We conclude that most VP1 modification occurs before viral DNA encapsidation, and that one defect in hr-t mutant virus assembly is in VP1 phosphorylation, primarily affecting threonine.
The effect of type of physical activity measure on the relationship between body fatness and habitual physical activity in children: a meta-analysis.

BACKGROUND The relationship between activity levels and body fat in children is unclear, despite a large number of studies. The issue is clouded by the wide variety of methods used to assess children's activity levels. It is important to assess whether the type of activity measure influences the fatness-activity relationship. This is a first step to uncovering the role of modifying variables such as gender, age, maturity, etc. PRIMARY OBJECTIVE This study uses meta-analytic procedures to synthesize the results of such studies and to assess whether the type of activity measure used has an effect on the strength of the relationship observed. METHODS AND PROCEDURES Fifty studies were located that satisfied the inclusion criteria. Seventy-eight per cent of the studies showed a negative relationship, 18% no relationship and 4% a positive relationship between physical activity and body fatness. Data were analysed using the meta-analytic procedures described by Rosenthal (Meta-analytic Procedures for Social Research, Sage, 1991). MAIN OUTCOMES AND RESULTS The mean effect size indicated a small to moderate, inverse relationship (r = -0.16). Mean effect sizes differed significantly (F(3,52) = 8.04, p < 0.001) according to the activity measure used: questionnaire, r = -0.14; motion counters, r = -0.18; observation, r = -0.39; heart rate (HR), r = 0.00. Observation measures elicited a significantly stronger relationship with body fat than did questionnaire or heart rate measures (p < 0.05). However, there was no significant difference between the effect sizes elicited by observation and motion counters. Correlational analyses revealed no effect of age group or gender on the strength of the relationship between fatness and activity. CONCLUSIONS This meta-analysis suggests there is a small to moderate relationship between body fat and activity in children. It is important to note, however, that the size of the relationship depends on the activity measure used. It is therefore recommended that direct measures of movement, such as observation or motion counter methods, are used to assess the relationship of activity levels with health.
Using a user experience evaluation framework for eModeration

The use of eModeration (of examination scripts) can improve the efficiency of the examination moderation, while simultaneously lowering the risk of misplacing or delaying the moderation process. Despite the potential benefits of using an eModerate system in terms of optimising examination procedures, the implementation and application of such online moderation systems in the South African context is limited. Various factors could be contributing towards the resistance to the implementation and adoption of eModerate systems in higher education institutions. These factors include human factors as well as technical and organisational resistance to change. This study focuses on the human factors involved in eModeration (user experience) and attempts to answer the following research question: How can the User Experience Evaluation Framework for eModeration be utilised within the context of higher education institutions in South Africa? The research used a Design Science Research methodology, which included the design, development as well as testing of the User Experience Evaluation Framework for eModeration. This paper will report on identified issues pertaining to the User Experience Evaluation Framework for eModeration during the evaluation phase. The research was conducted at Midrand Graduate Institute (MGI), now trading as Pearson Institute of Higher Education, a private higher education institute in South Africa. The data generation methods included interviews with eModerators from different faculties within a private higher education institution. This paper makes a theoretical contribution to this area of study by identifying the problems that users might have with the implementation of the User Experience Evaluation Framework for eModeration as well as providing some insights into the user experience of eModerators.
Factors Associated With Shift Work Disorder in Nurses Working With Rapid-Rotation Schedules in Japan: The Nurses’ Sleep Health Project

Workers who meet the criteria for shift work disorder (SWD) have elevated levels of risk for various health and behavioral problems. However, the impact of having SWD on shiftworkers engaged in rapid-rotation schedules is unknown. Moreover, the risk factors for the occurrence of SWD remain unclear. To clarify these issues, we conducted a questionnaire-based, cross-sectional survey on a sample of shiftworking nurses. Responses were obtained from 1202 nurses working at university hospitals in Tokyo, Japan, including 727 two-shift workers and 315 three-shift workers. The questionnaire included items relevant to age, gender, family structure, work environment, health-related quality of life (QOL), diurnal type, depressive symptoms, and SWD. Participants who reported insomnia and/or excessive sleepiness for at least 1 mo that was subjectively relevant to their shiftwork schedules were categorized as having SWD. The prevalence of SWD in the sampled shiftworking nurses was 24.4%; shiftworking nurses with SWD showed lower health-related QOL and more severe depressive symptoms, with greater rates of both actual accidents/errors and near misses, than those without SWD. The results of logistic regression analyses showed that more time spent working at night, frequent missing of nap opportunities during night work, and having an eveningness-oriented chronotype were significantly associated with SWD. The present study indicated that SWD might be associated with reduced health-related QOL and decreased work performance in shiftworking nurses on rapid-rotation schedules. The results also suggested that missing napping opportunities during night work, long nighttime working hours, and the delay of circadian rhythms are associated with the occurrence of SWD among shiftworking nurses on rapid-rotation schedules. (Author correspondence: yuinoue@tokyo-med.ac.jp)
The Mcf1 toxin induces apoptosis via the mitochondrial pathway and apoptosis is attenuated by mutation of the BH3‐like domain

Photorhabdus are Gram‐negative, nematode‐vectored bacteria that produce toxins to kill their insect hosts. The expression of one of these, Makes caterpillars floppy 1 (Mcf1), is sufficient to allow Escherichia coli to survive within, and kill, caterpillars which are otherwise able to clear E. coli infection. Mcf1 treated caterpillars show rapid loss of body turgor (the ‘floppy’ phenotype) and death is associated with massive apoptosis of both the midgut epithelium and insect phagocytes. Mammalian tissue culture cells treated with Mcf1 also display key features of apoptosis including zeiosis, apoptotic nuclear morphology, DNA laddering, activation of the effector caspase‐3 and PARP cleavage. As Mcf1 carries a single BH3‐like domain, here we investigate the hypothesis that this toxin promotes apoptosis via the mitochondrial pathway by mimicking a BH3 domain‐only protein. Consistent with this hypothesis, a double mutant within the BH3‐like domain causes a dramatic decline in apoptosis. Mcf1 also alters mitochondrial membrane potential and triggers the release of cytochrome c. Cells overexpressing Bcl‐xL, an anti‐apoptotic Bcl‐2 family member, are resistant to Mcf1‐mediated apoptosis, as are cells deficient in Bax. In addition, translocation of Bax to the mitochondrion is observed in response to Mcf1 treatment. Together, these results show that Mcf1 mediates apoptosis via the mitochondrial pathway, and are consistent with the hypothesis that the BH3‐like domain in Mcf1 is a functional requirement for the pro‐apoptotic activity of Mcf1.
Salvage endoscopic submucosal dissection in patients with local failure after chemoradiotherapy for esophageal squamous cell carcinoma

Abstract Objective. For locoregional failure after chemoradiotherapy (CRT) in patients with esophageal squamous cell carcinoma (ESCC), salvage esophagectomy and endoscopic mucosal resection have disadvantages, such as a high morbidity rate and a high local recurrence rate, respectively. The aim of this study was to clarify the efficacy of salvage endoscopic submucosal dissection (ESD) for locoregional failure of CRT. Methods. A total of 19 lesions in 19 patients were treated with salvage ESD; 15 lesions were local recurrences at the primary site and 4 lesions were residual. All lesions were intramucosal or submucosal tumors without metastases. A case-control study was retrospectively evaluated to clarify whether the clinical outcomes of salvage ESD were equivalent to those of control primary ESD. Results. No significant differences were observed between salvage ESD and primary ESD in short-term outcomes, including procedure time. For salvage ESD, the complete en bloc resection rate was 94.7% (18 of 19), and no severe complications were observed. At a median follow up of 54.6 (range: 5–98) months after salvage ESD, the local recurrence rate was 0%. However, three patients (15.8%) died due to lymph node and distant metastases and six patients (31.5%) died from other diseases, including radiation pneumonitis, pyothorax or respiratory failure with no recurrence of ESCC. The 3-year overall survival rate for all 19 patients was 74%. Conclusions. ESD represents an acceptable treatment option for recurrent or residual ESCC because of its improvement in local control, when local failure after CRT is limited to the submucosal layer without metastases.
Rap1a Null Mice Have Altered Myeloid Cell Functions Suggesting Distinct Roles for the Closely Related Rap1a and 1b Proteins1

The Ras-related GTPases Rap1a and 1b have been implicated in multiple biological events including cell adhesion, free radical production, and cancer. To gain a better understanding of Rap1 function in mammalian physiology, we deleted the Rap1a gene. Although loss of Rap1a expression did not initially affect mouse size or viability, upon backcross into C57BL/6J mice some Rap1a−/− embryos died in utero. T cell, B cell, or myeloid cell development was not disrupted in Rap1a −/− mice. However, macrophages from Rap1a null mice exhibited increased haptotaxis on fibronectin and vitronectin matrices that correlated with decreased adhesion. Chemotaxis of lymphoid and myeloid cells in response to CXCL12 or CCL21 was significantly reduced. In contrast, an increase in FcR-mediated phagocytosis was observed. Because Rap1a was previously copurified with the human neutrophil NADPH oxidase, we addressed whether GTPase loss affected superoxide production. Neutrophils from Rap1a−/− mice had reduced fMLP-stimulated superoxide production as well as a weaker initial response to phorbol ester. These results suggest that, despite 95% amino acid sequence identity, similar intracellular distribution, and broad tissue distribution, Rap1a and 1b are not functionally redundant but rather differentially regulate certain cellular events.
Thermoregulation and pain perception: Evidence for a homoeostatic (interoceptive) dimension of pain

Experimental and clinical observations of interactions between the nociceptive and thermoceptive systems have suggested that they could be part of the homoeostatic system relating to the condition of the body, described as ‘interoception’. Homoeostatic physiological systems are extensively interconnected. Thus, consistent with this hypothesis, we would expect thermoregulatory challenges to be associated with changes in pain perception.
N4ITK: Improved N3 Bias Correction

A variant of the popular nonparametric nonuniform intensity normalization (N3) algorithm is proposed for bias field correction. Given the superb performance of N3 and its public availability, it has been the subject of several evaluation studies. These studies have demonstrated the importance of certain parameters associated with the B-spline least-squares fitting. We propose the substitution of a recently developed fast and robust B-spline approximation routine and a modified hierarchical optimization scheme for improved bias field correction over the original N3 algorithm. Similar to the N3 algorithm, we also make the source code, testing, and technical documentation of our contribution, which we denote as ¿N4ITK,¿ available to the public through the Insight Toolkit of the National Institutes of Health. Performance assessment is demonstrated using simulated data from the publicly available Brainweb database, hyperpolarized 3He lung image data, and 9.4T postmortem hippocampus data.
Type I receptor binding of bone morphogenetic protein 6 is dependent on N‐glycosylation of the ligand

Bone morphogenetic proteins (BMPs), together with transforming growth factor (TGF)‐β and activins/inhibins, constitute the TGF‐β superfamily of ligands. This superfamily is formed by more than 30 structurally related secreted proteins. The crystal structure of human BMP‐6 was determined to a resolution of 2.1 Å; the overall structure is similar to that of other TGF‐β superfamily ligands, e.g. BMP‐7. The asymmetric unit contains the full dimeric BMP‐6, indicating possible asymmetry between the two monomeric subunits. Indeed, the conformation of several loops differs between both monomers. In particular, the prehelix loop, which plays a crucial role in the type I receptor interactions of BMP‐2, adopts two rather different conformations in BMP‐6, indicating possible dynamic flexibility of the prehelix loop in its unbound conformation. Flexibility of this loop segment has been discussed as an important feature required for promiscuous binding of different type I receptors to BMPs. Further studies investigating the interaction of BMP‐6 with different ectodomains of type I receptors revealed that N‐glycosylation at Asn73 of BMP‐6 in the wrist epitope is crucial for recognition by the activin receptor type I. In the absence of the carbohydrate moiety, activin receptor type I‐mediated signaling of BMP‐6 is totally diminished. Thus, flexibility within the binding epitope of BMP‐6 and an unusual recognition motif, i.e. an N‐glycosylation motif, possibly play an important role in type I receptor specificity of BMP‐6.
Constitutive Nuclear Localization of NFAT in Foxp3+ Regulatory T Cells Independent of Calcineurin Activity

Foxp3 plays an essential role in conferring suppressive functionality to CD4+/Foxp3+ regulatory T cells (Tregs). Although studies showed that Foxp3 has to form cooperative complexes with NFAT to bind to target genes, it remains unclear whether NFAT is available in the nucleus of primary Tregs for Foxp3 access. It is generally believed that NFAT in resting cells resides in the cytoplasm, and its nuclear translocation depends on calcineurin (CN) activation. We report that a fraction of NFAT protein constitutively localizes in the nucleus of primary Tregs, where it selectively binds to Foxp3 target genes. Treating Tregs with CN inhibitor does not induce export of NFAT from the nucleus, indicating that its nuclear translocation is independent of CN activity. Consistently, Tregs are resistant to CN inhibitors in the presence of IL-2 and continue to proliferate in response to anti-CD3 stimulation, whereas proliferation of non-Tregs is abrogated by CN inhibitors. In addition, PMA, which activates other transcription factors required for T cell activation but not NFAT, selectively induces Treg proliferation in the absence of ionomycin. TCR interaction with self-MHC class II is not required for PMA-induced Treg proliferation. Tregs expanded by PMA or in the presence of CN inhibitors maintain Treg phenotype and functionality. These findings shed light on Treg biology, paving the way for strategies to selectively activate Tregs.
Predictive values of body mass index and waist circumference for risk factors of certain related diseases in Chinese adults--study on optimal cut-off points of body mass index and waist circumference in Chinese adults.

OBJECTIVE For prevention of obesity in Chinese population, it is necessary to define the optimal range of healthy weight and the appropriate cut-off points of BMI and waist circumference for Chinese adults. The Working Group on Obesity in China under the support of International Life Sciences Institute Focal point in China organized a meta-analysis on the relation between BMI, waist circumference and risk factors of related chronic diseases (e.g., high diabetes, diabetes mellitus, and lipoprotein disorders). METHODS 13 population studies in all met the criteria for enrollment, with data of 239,972 adults (20-70 year) surveyed in the 1990s. Data on waist circumference was available for 111,411 persons and data on serum lipids and glucose were available for more than 80,000. The study populations located in 21 provinces, municipalities and autonomous regions in mainland China as well as in Taiwan. Each enrolled study provided data according to a common protocol and uniform format. The Center for data management in Department of Epidemiology, Fu Wai Hospital was responsible for statistical analysis. RESULTS AND CONCLUSION The prevalence of hypertension, diabetes, dyslipidemia and clustering of risk factors all increased with increasing levels of BMI or waist circumference. BMI at 24 with best sensitivity and specificity for identification of the risk factors, was recommended as the cut-off point for overweight, BMI at 28 which may identify the risk factors with specificity around 90% was recommended as the cut-off point for obesity. Waist circumference beyond 85 cm for men and beyond 80 cm for women were recommended as the cut-off points for central obesity. Analysis of population attributable risk percent illustrated that reducing BMI to normal range (< 24) could prevent 45%-50% clustering of risk factors. Treatment of obese persons (BMI > or = 28) with drugs could prevent 15%-17% clustering of risk factors. The waist circumference controlled under 85 cm for men and under 80 cm for women, could prevent 47%-58% clustering of risk factors. According to these, a classification of overweight and obesity for Chinese adults is recommended.