Anti-interleukin-1 treatment in patients with rheumatoid arthritis and type 2 diabetes (TRACK): A multicentre, open-label, randomised controlled trial.
The inflammatory contribution to type 2 diabetes (T2D) has suggested new therapeutic targets using biologic drugs designed for rheumatoid arthritis (RA). On this basis, we aimed at investigating whether interleukin-1 (IL-1) inhibition with anakinra, a recombinant human IL-1 receptor antagonist, could improve both glycaemic and inflammatory parameters in participants with RA and T2D compared with tumour necrosis factor (TNF) inhibitors (TNFis). This study, designed as a multicentre, open-label, randomised controlled trial, enrolled participants, followed up for 6 months, with RA and T2D in 12 Italian rheumatologic units between 2013 and 2016. Participants were randomised to anakinra or to a TNFi (i.e., adalimumab, certolizumab pegol, etanercept, infliximab, or golimumab), and the primary end point was the change in percentage of glycated haemoglobin (HbA1c%) (EudraCT: 2012-005370-62 ClinicalTrial.gov: NCT02236481). In total, 41 participants with RA and T2D were randomised, and 39 eligible participants were treated (age 62.72 ± 9.97 years, 74.4% female sex). The majority of participants had seropositive RA disease (rheumatoid factor and/or anticyclic citrullinated peptide antibody [ACPA] 70.2%) with active disease (Disease Activity Score-28 [DAS28]: 5.54 ± 1.03; C-reactive protein 11.84 ± 9.67 mg/L, respectively). All participants had T2D (HbA1c%: 7.77 ± 0.70, fasting plasma glucose: 139.13 ± 42.17 mg). When all the enrolled participants reached 6 months of follow-up, the important crude difference in the main end point, confirmed by an unplanned ad interim analysis showing the significant effects of anakinra, which were not observed in the other group, led to the study being stopped for early benefit. Participants in the anakinra group had a significant reduction of HbA1c%, in an unadjusted linear mixed model, after 3 months (β: -0.85, p < 0.001, 95% CI -1.28 to -0.42) and 6 months (β: -1.05, p < 0.001, 95% CI -1.50 to -0.59). Similar results were observed adjusting the model for relevant RA and T2D clinical confounders (male sex, age, ACPA positivity, use of corticosteroids, RA duration, T2D duration, use of oral antidiabetic drug, body mass index [BMI]) after 3 months (β: -1.04, p < 0.001, 95% CI -1.52 to -0.55) and 6 months (β: -1.24, p < 0.001, 95% CI -1.75 to -0.72). Participants in the TNFi group had a nonsignificant slight decrease of HbA1c%. Assuming the success threshold to be HbA1c% ≤ 7, we considered an absolute risk reduction (ARR) = 0.42 (experimental event rate = 0.54, control event rate = 0.12); thus, we estimated, rounding up, a number needed to treat (NNT) = 3. Concerning RA, a progressive reduction of disease activity was observed in both groups. No severe adverse events, hypoglycaemic episodes, or deaths were observed. Urticarial lesions at the injection site led to discontinuation in 4 (18%) anakinra-treated participants. Additionally, we observed nonsevere infections, including influenza, nasopharyngitis, upper respiratory tract infection, urinary tract infection, and diarrhoea in both groups. Our study has some limitations, including open-label design and previously unplanned ad interim analysis, small size, lack of some laboratory evaluations, and ongoing use of other drugs. In this study, we observed an apparent benefit of IL-1 inhibition in participants with RA and T2D, reaching the therapeutic targets of both diseases. Our results suggest the concept that IL-1 inhibition may be considered a targeted treatment for RA and T2D. The trial is registered with EU Clinical Trials Register, EudraCT Number: 2012-005370-62 and with ClinicalTrial.gov, number NCT02236481.


Bigenic Cre/loxP, puDeltatk conditional genetic ablation.
Genetic ablation experiments are used to resolve problems regarding cell lineages and the in vivo function of certain groups of cells. We describe a two-component conditional ablation technology using a mouse carrying an X-linked puDeltatk transgene, which is only activated in cells expressing Cre. Ablation of the Cre-expressing cells can be temporally regulated by the time of ganciclovir (GCV) administration. This strategy was demonstrated using a Col2Cre transgenic line. Differentiating chondrocytes in bigenic animals could be ablated at different developmental stages resulting in disorganized growth plates and dwarfism. Macrocephaly, macroglossia and umbilical hernia were also observed in ablated 18.5 dpc embryos. Crosses between the puDeltatk selector transgenic line and existing cre lines will facilitate numerous temporally regulated tissue-specific ablation experiments.


Is the National Guideline Clearinghouse a Trustworthy Source of Practice Guidelines for Child and Youth Anxiety and Depression?
Innovative strategies that facilitate the use of high quality practice guidelines (PG) are needed. Accordingly, repositories designed to simplify access to PGs have been proposed as a critical component of the network of linked interventions needed to drive increased PG implementation. The National Guideline Clearinghouse (NGC) is a free, international online repository. We investigated whether it is a trustworthy source of child and youth anxiety and depression PGs. English language PGs published between January 2009 and February 2016 relevant to anxiety or depression in children and adolescents (≤ 18 years of age) were eligible. Two trained raters assessed PG quality using Appraisal of Guidelines for Research and Evaluation (AGREE II). Scores on at least three AGREE II domains (stakeholder involvement, rigor of development, and editorial independence) were used to designate PGs as: i) minimum quality (≥ 50%); and ii) high quality (≥ 70%). Eight eligible PGs were identified (depression, n=6; anxiety and depression, n=1; social anxiety disorder, n=1). Four of eight PGs met minimum quality criteria; three of four met high quality criteria. At present, NGC users without the time and special skills required to evaluate PG quality may unknowingly choose flawed PGs to guide decisions about child and youth anxiety and depression. The recent NGC decision to explore the inclusion of PG quality profiles based on Institute of Medicine standards provides needed leadership that can strengthen PG repositories, prevent harm and wasted resources, and build PG developer capacity.


Comparative effects of theophylline and adenosine on respiratory skeletal and smooth muscle.
This study compared the effects of theophylline and adenosine on the contractile activity of respiratory smooth and skeletal muscles in vitro. Studies were performed on isolated strips of tracheal smooth and diaphragmatic skeletal muscle from 22 Hartley strain guinea pigs. The changes in diaphragm and tracheal smooth muscle tension in response to electrical stimulation and histamine-induced contraction, respectively, were assessed over a range of theophylline and adenosine concentrations (10(-7) to 10(-3) M for each). Theophylline increased diaphragmatic tension and reversed histamine-induced smooth muscle contractions in a dose-dependent fashion. Theophylline, however, decreased tracheal tension at concentrations an order of magnitude lower than those that increased diaphragmatic tension. Adenosine mimicked the effect of theophylline on tracheal muscle but had no effect on the force of diaphragm contraction. Moreover, adenosine failed to alter the theophylline dose-response curve for diaphragmatic tension. This study demonstrates that airway smooth muscle is more sensitive than is respiratory skeletal muscle to the effects of theophylline. Furthermore, these data suggest that the effects of theophylline on diaphragm skeletal muscle are not mediated by adenosine receptor blockade.


Oligomeric proanthocyanidin complexes: history, structure, and phytopharmaceutical applications.
Considerable recent research has explored therapeutic applications of oligomeric proanthocyanidin complexes (OPCs), naturally occurring plant metabolites widely available in fruits, vegetables, nuts, seeds, flowers, and bark. OPCs are primarily known for their antioxidant activity. However, these compounds have also been reported to demonstrate antibacterial, antiviral, anticarcinogenic, anti-inflammatory, anti-allergic, and vasodilatory actions. In addition, they have been found to inhibit lipid peroxidation, platelet aggregation, capillary permeability and fragility, and to affect enzyme systems including phospholipase A2, cyclooxygenase, and lipoxygenase. Based on these reported findings, OPCs may be a useful component in the treatment of a number of conditions.


Lipid peroxidation by Pseudomonas aeruginosa in the pathogenesis of nosocomial sepsis.
To study whether Pseudomonas aeruginosa may directly trigger peroxidation of polyunsaturated fatty acids, since lipid peroxidation is a mechanism involved in the pathogenesis of sepsis. Gamma-linolenic acid (GLA) was administered intravenously at a dose of 25mg/kg in an infusion time of 10 minutes to seven male rabbits. Blood samples were collected from the hepatic veins and from the carotid artery at regular time intervals. One clinical isolate was ex vivo incubated with the serum derived from the latter samples and concentrations of malondialdehyde (MDA) were determined during incubation in the growth medium by the thiobarbiturate assay. Elevated concentrations of MDA compared to their basal levels were found over the first three hours of incubation in the presence of samples collected 30 to 60 minutes after the end of the infusion of GLA. After infusion of GLA concentrations of arachidonic acid in the serum increased to concentrations comparable to those detected in sepsis. Direct triggering of lipid peroxidation by nosocomial isolates might be proposed as a pathogenetic mechanism of sepsis.


The impact of the COVID-19 pandemic on the number of presentations of penetrating injuries to a UK major trauma centre.
Knife-related violence is of growing concern in the UK. This study aims to investigate the impact of the COVID-19 pandemic on the frequency of penetrating injuries at a UK major trauma centre. This was a retrospective study comparing the number of patients attending the emergency department of King's College Hospital (KCH) with a penetrating injury (gunshot or stab wound) during the 'pandemic year' (1 March 2020-28 February 2021) compared with the equivalent time period in the previous year. Penetrating injuries as a result of self-harm were excluded. The primary outcome was to assess whether there were any changes to the frequency of presentations during three periods of national lockdowns. Lockdown 1 showed a 48.45% reduction in presentations in the 'pandemic year' compared to the previous year, lockdown 2 showed a 31.25% reduction; however, lockdown 3 showed an 8.89% increase in the number of presentations. Our findings suggest that despite the initial reduction in the number of presentations of penetrating injury during lockdown 1, this returned to normal levels by lockdown 3. Further research is required to understand the effects of government-imposed restrictions on interpersonal violence and identify appropriate methods of outreach prevention during a pandemic.


Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production.
Cellulosic biofuel is the subject of increasing attention. The main obstacle toward its economic feasibility is the recalcitrance of lignocellulose requiring large amount of enzyme to break. Several engineered yeast strains have been developed with cellulolytic activities to reduce the need for enzyme addition, but exhibiting limited effect. Here, we report the successful engineering of a cellulose-adherent Saccharomyces cerevisiae displaying four different synergistic cellulases on the cell surface. The cellulase-displaying yeast strain exhibited clear cell-to-cellulose adhesion and a "tearing" cellulose degradation pattern; the adhesion ability correlated with enhanced surface area and roughness of the target cellulose fibers, resulting in higher hydrolysis efficiency. The engineered yeast directly produced ethanol from rice straw despite a more than 40% decrease in the required enzyme dosage for high-density fermentation. Thus, improved cell-to-cellulose interactions provided a novel strategy for increasing cellulose hydrolysis, suggesting a mechanism for promoting the feasibility of cellulosic biofuel production.


[Optimizing the bone cement-implant interface by hydrolysis-resistant conditioning of the metal surface].
The hydrolytic degradation of the implant-cement interface has to be seen as the main reason for aseptic loosening of cemented total hip replacements. Therefore, a new method of conditioning the metallic surface was developed in order to achieve a hydrolytic-resistant bound stability between the implant and bone cement. Preliminary experimental data on test bodies are presented here. The metallic surface of 6 pairs of cylindrical test bodies each (CoCr-alloy, circular testing surface with O 6 mm) were conditioned by the method of silicoating/silanisation to gain a covalent coupling with the applied bone cement. In order to examine the initial stability and the hydrolytic resistance of the metal-cement compound, these pairs of surface-conditioned test bodies (SCT) as well as a reference series of surface-unconditioned test bodies (SUT) were immersed for 0, 30, 90, 150 days (d) in moisture environment (physiological saline solution, 37 degrees C) after coupling with bone cement. The adhesive strength of the test bodies-(bone cement-compounds) were determined by tensile tests on an universal testing machine (Typ Z030, Zwick, Ulm) with gimbal suspension. At time 0 d (that was without immersion of the test bodies) the mean maximum tensile bond strength of the SCT-cement-compounds was 39.5 MPa (SD +/- 4.7 MPa) and that of the SUT-cement-compounds 37.1 MPa (SD +/- 7.3 MPa) (p = 0.575). After immersion the tensile bond strength of the SUT-cement-compounds significantly decreased to an average of 13.5 MPa (SD +/- 2.7 MPa) (30 d), 10 MPa (SD +/- 1.7 MPa) (90 d) and 12.3 MPa (SD +/- 1.4 MPa) (150 d) (p < 0.01). In contrast, the SCT-cement-compounds showed a nearly unchanged high mechanical stability with tensile bond strength values of 37.0 MPa (SD +/- 4.9 MPa) after 30 d, 36.1 MPa (SD +/- 5.0 MPa) after 90 d und 30.2 MPa (SD +/- 4.7 MPa) after 150 d (p > 0.01). With reservation as to further in vitro and in vivo investigations the increased hydrolytic stability of the metal-cement-bound of surface-conditioned CoCr-alloy test bodies promises an improvement of the long-term stability of cement total joint replacements.


Discovery and horizontal follow-up of an autoantibody signature in human prostate cancer.
In response to an urgent need for improved diagnostic and predictive serum biomarkers for management of metastatic prostate cancer, we used phage display fingerprinting to analyze sequentially acquired serum samples from a patient with advancing prostate cancer. We identified a peptide ligand, CTFAGSSC, demonstrating an increased recovery frequency over time. Serum antibody reactivity to this peptide epitope increased in the index patient, in parallel with development of deteriorating symptoms. The antigen mimicking the peptide epitope was identified as alpha-2-Heremans-Schmid glycoprotein, also known as fetuin-A. Metastatic prostate cancer cell lines and bone metastasis samples displayed robust fetuin-A expression, and we demonstrated serum immune reactivity to fetuin-A with concomitant development of metastatic castrate-resistant disease in a large cohort of prostate cancer patients. Whereas fetuin-A is an established tumor antigen in several types of cancer, including breast cancer, glioblastoma, and pancreas cancer, this report is to our knowledge the first study implicating fetuin-A in prostate cancer and indicating that autoantibodies specific for fetuin-A show utility as a prognostic indicator for prostate cancer patients prone to progress to metastatic disease.


Construction of a physical fitness evaluation index system and model for high-level freestyle skiing aerials athletes in China.
This study aims to enhance the competitive level of Chinese freestyle skiing aerials athletes by developing a specialized physical fitness evaluation index system and model tailored for high-level Chinese athletes. This system intends to provide theoretical references and training monitoring schemes in preparation for the 25th Milan Winter Olympics. A study was conducted on 29 high-level Chinese freestyle skiing aerials athletes. Physical fitness test indexes were selected using a literature review, expert interviews, and questionnaire surveys, and athletes were tested. Athletes were ensured to be in optimal physical condition before testing. Based on the test results, the representative indexes of the evaluation system are finally determined by combining R-type clustering analysis, multiple linear regression analysis. Determine index weights through weight questionnaires and normalization, and develop evaluation standards through methods such as percentile counting and weighted scoring. Physical fitness evaluation system for Chinese freestyle skiing aerialists includes three aspects: evaluation index, index weight, and evaluation standard. The evaluation indexes include 3 first-level, 11 second-level, and 11 third-level indexes of body form, physiological function, and physical quality. In the evaluation weight, physical quality is ranked first, and physiological function and body form rank second and third, respectively. The evaluation standard consists of a scoring evaluation standard and a rating evaluation standard. Based on the index system, this study constructs the general and ideal physical fitness model of China's high-level freestyle aerials athletes. The constructed physical fitness evaluation system effectively represents physical fitness development status of high-level freestyle skiing aerials athletes, providing a basis for creating personalized training plans. The established model serves as a reference for athletes' physical fitness development objectives.


Role of implantable pacemakers in control of recurrent ventricular tachycardia.
Implanted pacemakers can provide a viable alternative to pharmacologic therapy or surgical management of patients with recurrent ventricular tachycardia. An increasing variety of pacemaker techniques are proving useful for preventing, controlling the effective rate of or terminating ventricular tachycardia. To assess the role of permanent pacing in the treatment of ventricular tachycardia, a consecutive series of 160 patients undergoing electrophysiologic testing for recurrent ventricular tachycardia were analyzed. Thirty-nine patients received implantable pacemakers, of which 13 were intended for bursts of rapid ventricular pacing. No adverse responses were attributed to these burst pacemakers, but concern for possible acceleration of tachycardia and appropriate identification of the arrhythmia were among factors limiting more widespread use of antitachycardia pacing. Some of these limitations may be resolved with further advances in electrophysiologic understanding of arrhythmias combined with strides in medical electronics, which will permit the development of new generations of highly sophisticated antitachycardia pacemakers.


Optimal Treatment of Retinal Vein Occlusion: Canadian Expert Consensus.
The availability of new therapeutic approaches, particularly intravitreal anti-vascular endothelial growth factor (anti-VEGF) therapies, has prompted significant changes to the established treatment paradigms for retinal vein occlusion (RVO). Better visual outcomes and significantly lower rates of adverse events have been noted in multiple large randomized clinical trials and have led to a new standard of care for this sight-threatening condition. To develop an expert consensus for the management of RVO and associated complications in the context of recent clinical evidence. The development of a Canadian expert consensus for optimal treatment began with a review of clinical evidence, daily practice, and existing treatment guidelines and algorithms. The expert clinicians (11 Canadian retina specialists) met on February 1, 2014, in Toronto to discuss their findings and to propose strategies for consensus. The result of this expert panel is a consensus proposal for Canadian ophthalmologists and retina specialists treating patients presenting with RVO. Treatment algorithms specific to branch and central RVO (BRVO and CRVO) were also developed. The consensus provides guidelines to aid clinicians in managing RVO and associated complications in their daily practice. In summary, laser remains the therapy of choice when neovascularization secondary to RVO is detected. Adjunctive anti-VEGF could be considered in managing neovascularization secondary to RVO in cases of vitreous hemorrhage. Intravitreal anti-VEGF should be considered for symptomatic visual loss associated with center-involving macular edema on optical coherence tomography. Patients with BRVO and a suboptimal response to anti-VEGF could be treated with grid laser, and those with CRVO and an inadequate response to anti-VEGF may be candidates for intravitreal steroids.


Learning From Clinical Consensus Diagnosis in India to Facilitate Automatic Classification of Dementia: Machine Learning Study.
The Harmonized Diagnostic Assessment of Dementia for the Longitudinal Aging Study in India (LASI-DAD) is the first and only nationally representative study on late-life cognition and dementia in India (n=4096). LASI-DAD obtained clinical consensus diagnosis of dementia for a subsample of 2528 respondents. This study develops a machine learning model that uses data from the clinical consensus diagnosis in LASI-DAD to support the classification of dementia status. Clinicians were presented with the extensive data collected from LASI-DAD, including sociodemographic information and health history of respondents, results from the screening tests of cognitive status, and information obtained from informant interviews. Based on the Clinical Dementia Rating (CDR) and using an online platform, clinicians individually evaluated each case and then reached a consensus diagnosis. A 2-step procedure was implemented to train several candidate machine learning models, which were evaluated using a separate test set for predictive accuracy measurement, including the area under receiver operating curve (AUROC), accuracy, sensitivity, specificity, precision, F1 score, and kappa statistic. The ultimate model was selected based on overall agreement as measured by kappa. We further examined the overall accuracy and agreement with the final consensus diagnoses between the selected machine learning model and individual clinicians who participated in the clinical consensus diagnostic process. Finally, we applied the selected model to a subgroup of LASI-DAD participants for whom the clinical consensus diagnosis was not obtained to predict their dementia status. Among the 2528 individuals who received clinical consensus diagnosis, 192 (6.7% after adjusting for sampling weight) were diagnosed with dementia. All candidate machine learning models achieved outstanding discriminative ability, as indicated by AUROC >.90, and had similar accuracy and specificity (both around 0.95). The support vector machine model outperformed other models with the highest sensitivity (0.81), F1 score (0.72), and kappa (.70, indicating substantial agreement) and the second highest precision (0.65). As a result, the support vector machine was selected as the ultimate model. Further examination revealed that overall accuracy and agreement were similar between the selected model and individual clinicians. Application of the prediction model on 1568 individuals without clinical consensus diagnosis classified 127 individuals as living with dementia. After applying sampling weight, we can estimate the prevalence of dementia in the population as 7.4%. The selected machine learning model has outstanding discriminative ability and substantial agreement with a clinical consensus diagnosis of dementia. The model can serve as a computer model of the clinical knowledge and experience encoded in the clinical consensus diagnostic process and has many potential applications, including predicting missed dementia diagnoses and serving as a clinical decision support tool or virtual rater to assist diagnosis of dementia.


Aflatoxins as risk factors for hepatocellular carcinoma in humans.
On a global basis, primary liver cancer (PLC) is a very prevalent form of cancer. Wide variation of PLC incidence in different areas of the world suggests the involvement of environmental factors in its etiology. Two major classes of risk factors have been identified. Extensive evidence indicates the importance of infection by the hepatitis B virus as a major risk factor for PLC. Because many organic chemicals induce liver cancer in experimental animals, those to which human exposure is known to occur are also of interest with respect to their possible involvement as risk factors for PLC. Particular emphasis has been placed on aflatoxins because of the frequency with which they occur as food contaminants, together with their potency as liver carcinogens for a large number of experimental animals, including subhuman primates. Other mycotoxins, notably sterigmatocystin and fumonisin, also are relatively potent carcinogens for the liver of animals, but little is known about human exposure to them. Epidemiological surveys carried out over the past 25 years in Asia and Africa have revealed a strong statistical association between aflatoxin ingestion and PLC incidence. The combined experimental and epidemiological evidence has led to designation of aflatoxins as human carcinogens according to International Agency for Cancer Research criteria. Collectively, current evidence strongly suggests that PLC is of multifactorial origin, with probable interactions between viral and chemical agents in populations concurrently exposed to both classes of risk factors. Recently developed methods that permit individual monitoring of aflatoxin exposure, hepatitis B virus infection, and genetic damage caused by these agents are being applied in the design of molecular and biochemical epidemiological studies of the etiology of the disease. Application of this methodology may contribute to elucidation of the relative importance of interacting etiological agents in different populations.


Psychosocial consequences of disordered eating attitudes in elite female figure skaters.
The purpose of this study was to test a model of the effects of athletes' eating disorders on coach, sport friend and parent social relationships and peer social acceptance through physical self-perceptions (i.e. perceived physical appearance and perceived physical ability). The sample comprised 199 elite female figure skaters. The data were analysed using structural equation modelling. Our results showed that the effects of athletes' eating disorders on social relationships were not mediated by physical self-perceptions. Athletes' eating disorders directly negatively influenced the quality of the parent-athlete relationship, thus confirming previous findings in young adults. Contrary to the literature, eating disorders were positively related to perceived physical ability. This paradoxical finding is discussed in relation to achievement theories and the particularity of the sport context.


Chemical and structural characterization of a model Post-Termination Complex (PoTC) for the ribosome recycling reaction: Evidence for the release of the mRNA by RRF and EF-G.
A model Post-Termination Complex (PoTC) used for the discovery of Ribosome Recycling Factor (RRF) was purified and characterized by cryo-electron microscopic analysis and biochemical methods. We established that the model PoTC has mostly one tRNA, at the P/E or P/P position, together with one mRNA. The structural studies were supported by the biochemical measurement of bound tRNA and mRNA. Using this substrate, we establish that the release of tRNA, release of mRNA and splitting of ribosomal subunits occur during the recycling reaction. Order of these events is tRNA release first followed by mRNA release and splitting almost simultaneously. Moreover, we demonstrate that IF3 is not involved in any of the recycling reactions but simply prevents the re-association of split ribosomal subunits. Our finding demonstrates that the important function of RRF includes the release of mRNA, which is often missed by the use of a short ORF with the Shine-Dalgarno sequence near the termination site.


Relationship among Glutamine, γ-Aminobutyric Acid, and Social Cognition in Autism Spectrum Disorders.
An imbalance of excitatory and inhibitory neurotransmission in autism spectrum disorder (ASD) has been proposed. We compared glutamate (Glu), glutamine (Gln), and γ-aminobutyric acid (GABA) levels in the anterior cingulate cortex (ACC) of 13 males with ASD and 14 typically developing (TD) males (ages 13-17), and correlated these levels with intelligence quotient (IQ) and measures of social cognition. Social cognition was evaluated by administration of the Social Responsiveness Scale (SRS) and the Reading the Mind in the Eyes Test (RMET). We acquired proton magnetic resonance spectroscopy ((1)H-MRS) data from the bilateral ACC using the single voxel point resolved spectroscopy sequence (PRESS) to quantify Glu and Gln, and Mescher-Garwood point-resolved spectroscopy sequence (MEGA-PRESS) to quantify GABA levels referenced to creatine (Cr). There were higher Gln levels (p=0.04), and lower GABA/Cre levels (p=0.09) in the ASD group than in the TD group. There was no difference in Glu levels between groups. Gln was negatively correlated with RMET score (rho=-0.62, p=0.001) and IQ (rho=-0.56, p=0.003), and positively correlated with SRS scores (rho=0.53, p=0.007). GABA/Cre levels were positively correlated with RMET score (rho=0.34, p=0.09) and IQ (rho=0.36, p=0.07), and negatively correlated with SRS score (rho=-0.34, p=0.09). These data suggest an imbalance between glutamatergic neurotransmission and GABA-ergic neurotransmission in ASD. Higher Gln levels and lower GABA/Cre levels were associated with lower IQ and greater impairments in social cognition across groups.


Prevention of hemolytic disease of the fetus and newborn: what have we learned from animal models?
This review aims to highlight recent advances in our understanding of how anti-red blood cell (RBC) antibodies prevent erythrocyte immunization with an emphasis on new murine models. New murine models with clinically relevant human erythrocyte antigens have been used to understand the alloimmunization process and its inhibition. The search to elucidate the mechanism of action of IgG-mediated inhibition of erythrocyte alloimmunization has provided new evidence in support of a potential role for epitope masking, immune deviation and/or antigen modulation in this process. In addition, recent evidence suggests that blends of monoclonal antibodies targeting nonoverlapping epitopes on the RBC surface can improve the efficacy of monoclonal antibodies approaching that of polyclonal IgG. Animal models with defined alloantigens have helped to identify important mechanistic components that lead to alloimmunization and its inhibition by IgG. A better understanding of the underlying mechanisms leading to hemolytic disease of the fetus and newborn is required to develop the most effective prevention strategies for future patients.


[Thromboembolic complications and permanent endocavitary stimulation catheters].
Based on four cases of phlebitis associated with permanent intravenous cardiac stimulation catheters, the authors review the pathophysiology, clinical signs, diagnosis and treatment of this disease. The insertion of the catheter causes varying degrees of radiologically detectable thrombosis in 30 to 50 per cent of cases. The most frequent clinical presentation is the typical picture of phlebitis of the upper limb due to axillo-subclavian thrombosis. However, clinical signs are only present in 0.5 to 5 per cent of cases (1.7 per cent of cases in the present series). The risk of this complication can not be predicted on the basis of delay after the implantation. The incidence is increased in patients with more than one stimulating electrode, but this disease is not confined to this group of patients. The thrombosis may affect the superior vena cava or the right cardiac cavities, when it may also be associated with pulmonary embolism, as in one of the cases in this series. One case with arteriolar involvement (phlegmatia cerulea) is also described. The diagnosis may require impedance rheoplethysmography, two dimensional echocardiography, isotope phlebography or contrast phlebography, which remains the most frequently used technique. The treatment is medical, consisting of intravenous or subcutaneous heparin. Fibrinolysis may be indicated in cases with recent superior vena cava thrombosis and or other complications. To our knowledge, no controlled studies have been performed to define the optimal treatment. Long term prophylactic anticoagulant therapy is useless. There is never any indication for removal and replacement of the catheter responsible or for the removal of useless catheters left in place in patients with multiple implants.


Prospective study in bullous pemphigoid: association of high serum anti-BP180 IgG levels with increased mortality and reduced Karnofsky score.
Bullous pemphigoid (BP) is a subepidermal blistering disease characterized by autoantibodies against the two hemidesmosomal proteins, BP180 (type XVII collagen) and BP230. The multicentre prospective BLISTER (Bullous Pemphigoid Steroids and Tetracyclines) trial randomized 253 patients with BP to compare the benefits and harms between initial treatment with doxycycline or prednisolone. To analyse distinct autoantibody profiles for the prediction of the disease course in a well-characterized cohort of BP sera. One hundred and forty-three patients of the BLISTER trial consented to participate in this serological study. Sera taken at baseline were analysed by (i) indirect immunofluorescence, (ii) anti-BP180 NC16A (16th noncollagenous domain) and anti-BP230 enzyme-linked immunosorbent assay and (iii) immunoblotting with various substrates. Results were then linked with clinical parameters including age, Karnofsky score, number of blisters, related adverse events and mortality. Disease activity correlated with immunoglobulin (Ig)G anti-BP180 levels but not with levels of anti-BP230 IgG and anti-BP180 IgE. High levels of both anti-BP180 IgG and anti-BP230 IgG were associated with a low Karnofsky score. The presence of anti-BP230 IgG was more frequent in older patients. Those with higher total IgE serum levels suffered from fewer adverse events. Higher IgG anti-BP180 levels were associated with an increased 1-year mortality rate. Analysis of the autoantibody profile is not only of diagnostic relevance but may also be helpful in predicting the course of the disease.


[Study on the occurrence of adverse health events in nursing homes : contributions from the Liège SENIOR cohort].
The SENIOR study followed a cohort of older people living in nursing homes for 8 years and examined their adverse health events. The results were analysed after 1, 3 and 8 years to identify predictive factors and improve care. After 1 year, residents with poorer motor and muscle function had a higher incidence of adverse health events such as death and falls. Sarcopenia predicted mortality, while poor physical performance was associated with falls. At 3 years, poor nutritional status and poor balance were important predictors of mortality, falls and loss of independence. At 8 years, younger age, higher body mass index, and good physical and cognitive performance were associated with longer survival. The study also examined the impact of the COVID-19 pandemic in nursing homes and found no significant association between frailty, nutrition, muscle strength and COVID-19. In conclusion, functional capacity and nutrition play a crucial role in predicting adverse events in nursing home residents. The results will guide public health policies and clinical interventions to improve quality of life.


Skin Bacterial Community Reorganization Following Metamorphosis of the Fire-Bellied Toad (Bombina orientalis).
In organisms with complex life histories, dramatic changes in microbial community structure may occur with host development and immune system maturation. Amphibian host susceptibility to diseases such as chytridiomycosis may be affected by the reorganization of skin microbial community structure that occurs during metamorphosis. We tracked changes in the bacterial communities inhabiting skin of Korean fire-bellied toads (Bombina orientalis) that we infected as tadpoles with different strains of Batrachochytrium dendrobatidis (Bd), the pathogenic fungus that causes chytridiomycosis. We found that B. orientalis undergoes a major change in skin bacterial community composition between 5 and 15 days following metamorphosis. Richness indices and phylogenetic diversity measures began to diverge earlier, between aquatic and terrestrial stages. Our results further reveal differences in skin bacterial community composition among infection groups, suggesting that the effect of Bd infection on skin microbiome composition may differ by Bd strain. Additional studies are needed to further investigate the structural and temporal dynamics of microbiome shifts during metamorphosis in wild and captive amphibian populations. Analyses of the ontogeny of microbiome shifts may contribute to an understanding of why amphibians vary in their susceptibility to chytridiomycosis.


Glycoconjugate saccharidic moieties of the exocrine and endocrine pancreas in the chick embryo, newborn and adult.
A battery of horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, ConA, LTA and UEAI) was used to study the distribution of glycoconjugate sugar residues in the exocrine and endocrine pancreas of chick embryos, 1-day-old chicks and adult animals. During the period of incubation considered here, the time of appearance and changes in the oligosaccharides were noted in the acinar cells, in the endocrine cells of the "light islets" and in the capillary endothelial cells. In the developing exocrine pancreas, the basolateral surface of the acinar cells reacted with PNA, WGA, LTA and ConA for the entire period of incubation while SBA, WGA, LTA and ConA reactivity was detected in the apical plasma membrane. Zymogen granule membrane reacted with PNA, WGA and LTA from the 12th day onward, while ConA reactivity was detectable from the 7th day onward. Owing to its early appearance, alpha-D-mannose, revealed by ConA lectin, might in some way contribute to the maturation of the zymogen granule. In the 1-day-old chick and in the adult the cellular surface reacted with WGA and ConA. The zymogen granule membrane reacted with PNA, WGA and ConA in the 1-day-old chick. In the adult, WGA and ConA reactivity was observed in the zymogen granule membrane, whereas at this site PNA reactivity was revealed only after neuraminidase digestion. The hypothesis formulated by others that granule membrane lectin reactivity is equal to the apical membrane reactivity, owing to exocytotic processes, is not consistent with our results in the chick embryos, the 1-day-old chick and the adults. The surface and the granules of the islet beta cells in different periods of incubation reacted with WGA, SBA and, after neuraminidase treatment, with PNA. The same reactivity was seen also in 1-day-old chick while in the adult the granules of the beta cells also reacted with ConA. These findings show the achievement of an almost adult-like state of glycosylation of the glycoconjugates in the beta cells at an early stages of development.


Can early vigour occur in combination with drought tolerance and efficient water use in rice genotypes?
Selection for early vigour can improve rice (Oryza sativa L.) seedlings' access to resources, weed competitiveness and yield. Little is known about the relationships between early vigour and drought tolerance. This study explored a panel of 176 rice genotypes in a controlled environment regarding a diversity of traits and trait combinations related to early vigour and water use under drought. The design excluded genotypic differences for root depth. We hypothesised that early vigour (as determined by biomass accumulation under well-watered conditions) was not independent from drought tolerance (determined by biomass accumulation maintenance under drought). Leaf size, developmental rate (DR) and tiller number contributed positively to shoot DW and leaf area, and thus vigour. Early vigour was negatively correlated with growth maintenance and water use efficiency under drought, suggesting tradeoffs. Three clusters of genotypes were identified based on the constitutive traits DR, specific leaf area, tiller number and leaf size. The less drought-tolerant cluster, mainly with lowland O. sativa indica rices, showed a sensitive transpiration response to the fraction of transpirable soil water; however, under well-watered conditions these genotypes were vigorous, with small leaves, high DR and high tillering. This experiment showed that the tradeoff between early vigour and drought tolerance was physiological and not a matter of access to water. The results are discussed with a view to identify drought adaptation strategies for crop improvement. Further improvement of multitrait phenotyping approaches is proposed.


[Evaluation of the sanitary-and-epidemiological hazard of solid garbage in Astana].
According to the national plan of actions on environmental protection, industrial garbage recycling is to be introduced in Almaty and Astana for the sustainable development of the Republic of Kazakhstan. Integrated assessment of the hazard of garage is made by the sanitary-and-chemical and sanitary-and-epidemiological indices to provide the hygienic and ecological reliability of a procedure for neutralization and utilization of solid garbage (SG). According to the data obtained, Astana SG Astana in summer is characterized by the high total level of bacterial contamination. The indices of microbial contamination of SG and soil near the dustbins correlate with the density of population and the maturity of an infrastructure. Comparison of the sanitary-and-epidemiological indices of different types of SG (wastes from housing facilities, wholesale and retail outlays, and education, culture, and entertainment institutions) revealed no significant differences. According to the sanitary-and-helmintological indices, the Astana soil should be classified as pure (noninvasive). Involvement of SG into industrial recycling should be accompanied by a hygienic assessment of the hazard of waste and the reliability of used technologies in the context of warning and on-going sanitary surveillance.


New insights into stop codon recognition by eRF1.
In eukaryotes, translation termination is performed by eRF1, which recognizes stop codons via its N-terminal domain. Many previous studies based on point mutagenesis, cross-linking experiments or eRF1 chimeras have investigated the mechanism by which the stop signal is decoded by eRF1. Conserved motifs, such as GTS and YxCxxxF, were found to be important for termination efficiency, but the recognition mechanism remains unclear. We characterized a region of the eRF1 N-terminal domain, the P1 pocket, that we had previously shown to be involved in termination efficiency. We performed alanine scanning mutagenesis of this region, and we quantified in vivo readthrough efficiency for each alanine mutant. We identified two residues, arginine 65 and lysine 109, as critical for recognition of the three stop codons. We also demonstrated a role for the serine 33 and serine 70 residues in UGA decoding in vivo. NMR analysis of the alanine mutants revealed that the correct conformation of this region was controlled by the YxCxxxF motif. By combining our genetic data with a structural analysis of eRF1 mutants, we were able to formulate a new model in which the stop codon interacts with eRF1 through the P1 pocket.


Changes in striatal activity and functional connectivity in a mouse model of Huntington's disease.
Hereditary Huntington's disease (HD) is associated with progressive motor, cognitive and psychiatric symptoms. A primary consequence of the HD mutation is the preferential loss of medium spiny projection cells with relative sparing of local interneurons in the striatum. In addition, among GABAergic striatal projection cells, indirect pathway cells expressing D2 dopamine receptors are lost earlier than direct pathway cells expressing D1 receptors. To test in vivo the functional integrity of direct and indirect pathways as well as interneurons in the striatum of male R6/1 transgenic mice, we assessed their c-Fos expression levels induced by a striatal-dependent cognitive task and compared them with age-matched wild-type littermates. We found a significant increase of c-Fos+ nuclei in the dorsomedial striatum, and this only at 2 months, when our HD mouse model is still pre-motor symptomatic, the increase disappearing with symptom manifestation. Contrary to our expectation, the indirect pathway projection neurons did not undergo any severer changes of c-Fos expression regardless of age in R6/1 mice. We also found a decreased activation of interneurons that express parvalbumin in the dorsomedial striatum at both presymptomatic and symptomatic ages. Finally, analysis of c-Fos expression in extended brain regions involved in the cognitive learning used in our study, demonstrates, throughout ages studied, changes in the functional connectivity between regions in the transgenic mice. Further analysis of the cellular and molecular changes underlying the transient striatal hyperactivity in the HD mice may help to understand the mechanisms involved in the disease onset.


Cryptosporidium baileyi infection associated with an outbreak of ocular and respiratory disease in otus owls (Otus scops) in a rehabilitation centre.
Cryptosporidiosis has been reported in more than 30 avian species worldwide. Although some cases of cryptosporidiosis have been described in captive birds of prey in the order Falconiformes, to date there have been no reports of the disease in wild raptors. Here we describe for first time an ocular and respiratory disease associated with Cryptosporidium baileyi in wild scops owl (Otus scops, order: Strigiformes). Sixteen otus owl fledglings born in the wild during the summer of 2008 were admitted to the Torreferrussa Wildlife Rehabilitation Centre (Catalonia, northern Spain) in July and August of the same year. In the middle of September, blepharoedema, conjunctival hyperaemia and mucopurulent ocular discharge were diagnosed unilaterally in 75% (12/16) of the birds and bilaterally in 25% (4/16). Moreover, five birds (31%) developed diffuse epithelial corneal oedema, one owl (6%) displayed mild anterior exudative uveitis and another developed rhinitis (6%). Two birds were euthanized because of the severity of disease. The histopathology demonstrated cryptosporidia-like structures in the conjunctival cells and in the nasal respiratory epithelium of one owl. Cryptosporidium spp. oocysts (6.5 to 7.0 x 5.0 to 5.5 microm) were identified by immunofluorescent antibody test (IFAT) in histological sections from eyelids, trachea and respiratory sinuses and in swab samples from the glottis, choanal slit and conjunctival sac. Polymerase chain reaction and DNA sequence analysis confirmed the presence of C. baileyi. Birds were treated orally with azithromycin (40 mg/kg) once a day for 15 days, and by the end of the treatment all owls tested negative for the parasites, by IFAT, and did not display further signs of disease.


Measurement of dynamic task related functional networks using MEG.
The characterisation of dynamic electrophysiological brain networks, which form and dissolve in order to support ongoing cognitive function, is one of the most important goals in neuroscience. Here, we introduce a method for measuring such networks in the human brain using magnetoencephalography (MEG). Previous network analyses look for brain regions that share a common temporal profile of activity. Here distinctly, we exploit the high spatio-temporal resolution of MEG to measure the temporal evolution of connectivity between pairs of parcellated brain regions. We then use an ICA based procedure to identify networks of connections whose temporal dynamics covary. We validate our method using MEG data recorded during a finger movement task, identifying a transient network of connections linking somatosensory and primary motor regions, which modulates during the task. Next, we use our method to image the networks which support cognition during a Sternberg working memory task. We generate a novel neuroscientific picture of cognitive processing, showing the formation and dissolution of multiple networks which relate to semantic processing, pattern recognition and language as well as vision and movement. Our method tracks the dynamics of functional connectivity in the brain on a timescale commensurate to the task they are undertaking.


Direct and sensitive detection of circulating miRNA in human serum by ligase-mediated amplification.
MicroRNAs (miRNA) involve in regulating different physiological processes whose dysregulation is associated with a wide range of diseases including cancers, diabetes and cardiovascular problems. Herein, we report a direct, sensitive and highly selective detection assay for circulating microRNA (miRNA). This detection strategy employs magnetic nanoparticles as the reaction platform which can not only allow online pre-concentration and selective separation but also integrates ligation reaction with amplification to enhance the sensitivity of the detection assay. With the presence of the target miRNA, the locked nucleic acid (LNA)-modified molecular beacon (MB) opens up, exposing the binding sites at two ends. The 3'- and 5'-end of the MB responsible for the attachment onto the magnetic nanoparticles, and reporting probe for the attachment of the pair of amplification probes respectively. The ligase ligate RNA to DNA enhance the amplification efficiency. Upon labelled with intercalating fluorophores (YOYO-1) on the hybrids, the quantification of the target miRNA was determined by measuring the fluorescence intensity. A detection limit of 314 fM was achieved with trace amount of sample consumption (~20 μL). As a proof of concept, miRNA-149 was chosen as the target miRNA. This assay is capable of discriminating single-base and reliably quantifying circulating miRNA-149 in both healthy and cancer patient's serums. The result obtained was comparable with that of quantitative reverse transcription polymerase chain reaction (qRT-PCR), suggesting that this direct and sensitive assay can be served as a promising, non-invasive tool for early diagnosis of breast cancer and colorectal cancer.


Immunomodulation using bacterial enterotoxins.
Immunologic unresponsiveness (tolerance) is a key feature of the mucosal immune system, and deliberate vaccination by a mucosal route can effectively induce immune suppression. However, some bacterial-derived proteins, e.g. cholera toxin and the heat labile toxin of Escherichia coli, are immunogenic and immunomodulatory at mucosal surfaces and can effectively adjuvant immune responses to codelivered bystander antigens. This review summarizes some of the structural and biological characteristics of these toxins and provides examples of how these properties have been exploited for tolerance induction and mucosal vaccine development.


Influence of demographic factors on the occurrence of motion artefacts in HR-pQCT.
The study shows a high incidence of motion artefacts in a central European population and a significant increase of those artefacts with higher age. These findings may impact on the design and conduct of future in vivo HR-pQCT studies or at least help to estimate the potential number of drop outs due to unusable image quality. Motion artefacts in high-resolution peripheral quantitative computed tomography (HR-pQCT) are challenging, as they introduce error into the resulting measurement data. The aim of this study was to assess the general occurrence of motion artefacts in healthy distal radius and to evaluate the influence of demographic factors. The retrospective study is based on 525 distal radius second-generation HR-pQCT scans of 95 patients. All stacks were evaluated by two experienced observers and graded according to the visual grading scale recommended by the manufacturer, ranging from grade 1 (no visible motion artefacts) to grade 5 (severe motion artefacts). Correlations between demographic factors and image quality were evaluated using a linear mixed effects model analysis. The average visual grading was 2.7 (SD ± 0.7). Age and severity of motion artefacts significantly correlated (p = 0.026). Patients aged 65 years or above had an average image quality between grades 1 and 3 in 72.7% of cases, while patients younger than 65 had an average image quality between grades 1 and 3 in 91.9% of cases. Gender, smoking behaviour, and handedness had no significant influence on motion artefacts. This study showed a high incidence of motion artefacts in a representative central European population, but also a significant increase of motion artefacts with higher age. This could impact further study designs by planning for a sufficiently large and if possible a more selective study population to gain a representative amount of high-quality image data.


Ethanol sensitizes NF-kappaB activation in pancreatic acinar cells through effects on protein kinase C-epsilon.
Although ethanol abuse is the most common cause of pancreatitis, the mechanism of alcohol's effect on the pancreas is not well understood. Previously, we demonstrated that in vitro ethanol treatment of pancreatic acinar cells augmented the CCK-8-induced activation of NF-kappaB, a key signaling system involved in the inflammatory response of pancreatitis. In the present study, we determine the role for individual PKC isoforms in the sensitizing effect of ethanol on NF-kappaB activation. Dispersed rat pancreatic acini were treated with and without ethanol and then stimulated with CCK-8; 100 nM CCK-8 caused both NF-kappaB and PKC-delta, -epsilon, and -zeta activation, whereas 0.1 nM CCK-8 did not increase PKC-epsilon, PKC-zeta, or NF-kappaB activity. CCK-8 (0.1 nM) did activate PKC-delta. PKC-epsilon activator alone did not cause NF-kappaB activation; however, together with 0.1 nM CCK-8, it caused NF-kappaB activation. Ethanol activated PKC-epsilon without affecting other PKC isoforms or NF-kappaB activity. Of note, stimulation of acini with ethanol and 0.1 nM CCK-8 resulted in the activation of PKC-delta, PKC-epsilon, and NF-kappaB. The NF-kappaB activation to 0.1 nM CCK-8 in ethanol-pretreated acini was inhibited by both PKC-delta inhibitor and PKC-epsilon inhibitor. Taken together, these results demonstrate the different modes of activation of PKC isoforms and NF-kappaB in acini stimulated with ethanol, high-dose CCK-8, and low-dose CCK-8, and furthermore suggest that activation of both PKC-epsilon and -delta is required for NF-kappaB activation. These results suggest that ethanol enhances the CCK-8-induced NF-kappaB activation at least in part through its effects on PKC-epsilon.


Missense variants in the N-terminal domain of the A isoform of FHF2/FGF13 cause an X-linked developmental and epileptic encephalopathy.
Fibroblast growth factor homologous factors (FHFs) are intracellular proteins which regulate voltage-gated sodium (Nav) channels in the brain and other tissues. FHF dysfunction has been linked to neurological disorders including epilepsy. Here, we describe two sibling pairs and three unrelated males who presented in infancy with intractable focal seizures and severe developmental delay. Whole-exome sequencing identified hemi- and heterozygous variants in the N-terminal domain of the A isoform of FHF2 (FHF2A). The X-linked FHF2 gene (also known as FGF13) has alternative first exons which produce multiple protein isoforms that differ in their N-terminal sequence. The variants were located at highly conserved residues in the FHF2A inactivation particle that competes with the intrinsic fast inactivation mechanism of Nav channels. Functional characterization of mutant FHF2A co-expressed with wild-type Nav1.6 (SCN8A) revealed that mutant FHF2A proteins lost the ability to induce rapid-onset, long-term blockade of the channel while retaining pro-excitatory properties. These gain-of-function effects are likely to increase neuronal excitability consistent with the epileptic potential of FHF2 variants. Our findings demonstrate that FHF2 variants are a cause of infantile-onset developmental and epileptic encephalopathy and underline the critical role of the FHF2A isoform in regulating Nav channel function.


Nutrition knowledge of sports physical therapists.
This descriptive study determined the nutrition knowledge of sports physical therapists. A Nutrition Knowledge Test was designed and mailed to 1000 members of the Sports Section of the American Physical Therapy Association. The 498 therapists who returned completed questionnaires achieved an overall mean knowledge score of 71.63%. Women scored significantly better than men; therapists in the 16+ years of experience and the 50+ years age groups scored less well. While there was a positive attitude toward nutrition, no correlation was found between attitude and nutrition knowledge. Thirty percent of the therapists reported receiving nutrition education in their physical therapy program. Thirty-two percent received nutrition information through continuing education. Those therapists with continuing education achieved significantly higher knowledge scores, particularly on the sports-related questions. The results of this study indicate that the nutrition knowledge of sports physical therapists can be increased with nutrition education provided through continuing education. J Orthop Sports Phys Ther 1989;10(7):257-263.


In vitro generation of a highly immunogenic subline of L1210 leukemia following exposure to 5-(3,3'-dimethyl-1-triazeno)imidazole-4-carboxamide.
Strong and heritable increase of immunogenicity of L1210 Ha leukemia has been obtained in vitro following multiple treatments with 5-(3,3'-dimethyl-1-triazeno)imidazole-4-carboxamide (DTIC), metabolically activated by mouse liver preparations (MLP) containing liver microsomes. The DTIC-treated leukemia (L1210D line) or the control line treated with MLP alone (L1210N line) showed comparable growth kinetics in vitro. However, progressive increase of immunogenicity occurred in leukemic cells in the course of in vitro treatments with DTIC plus MIP, but not with MLP alone, as evidenced by comparative studies on transplantation immunity elicited in BALB/c x DBA/2 F1 mice by graded inocula of L1210D or L1210N leukemia cells. In vitro experiments confirmed that metabolic transformation of DTIC is required for increasing tumor immunogenicity. In fact, L1210Ha cells became highly immunogenic when treated with DTIC in intact mice but not in animals metabolically depressed by CCl4. Immunochemotherapy experiments based on the antigenic cross-reactivity between the L1210D line and the original L1210Ha leukemia showed that i.p. administration of L1210D cells followed by 1,3-bis(2-chloroethyl)-1-nitrosourea treatment afforded marked protection in mice inoculated intracerebrally with the parental lymphoma. The present findings could provide an adequate in vitro technique for developing further studies on DTIC-mediated immunogenic changes of tumors, including human cancer cells growing in tissue culture.


Endogenous Myc maintains the tumor microenvironment.
The ubiquitous deregulation of Myc in human cancers makes it an intriguing therapeutic target, a notion supported by recent studies in Ras-driven lung tumors showing that inhibiting endogenous Myc triggers ubiquitous tumor regression. However, neither the therapeutic mechanism nor the applicability of Myc inhibition to other tumor types driven by other oncogenic mechanisms is established. Here, we show that inhibition of endogenous Myc also triggers ubiquitous regression of tumors in a simian virus 40 (SV40)-driven pancreatic islet tumor model. Such regression is presaged by collapse of the tumor microenvironment and involution of tumor vasculature. Hence, in addition to its diverse intracellular roles, endogenous Myc serves an essential and nonredundant role in coupling diverse intracellular oncogenic pathways to the tumor microenvironment, further bolstering its credentials as a pharmacological target.


Adjustable task lighting: Field study assesses the benefits in an office environment.
Lighting is a part of every work task in the office environment, yet it is often overlooked. Research links direct and indirect glare to increased risk of visual discomfort among office workers with symptoms ranging from dry eyes to blurry vision or headaches. Researchers have been primarily concerned with those characteristics of task lighting that cause glare including luminance level, position (line of sight), and control. It is unknown what the benefits of adjustable task lights are and whether or not their use has an effect on musculoskeletal comfort or posture. No comprehensive field evaluations of this type were found among peer-reviewed, indexed journals. The purpose of this study was to assess the ergonomic and calculated utility power consumption benefits of adjustable LED task lighting in an office environment using a control/intervention experiment design. One hundred participants were originally recruited and randomly assigned to intervention and control groups. Self-reported data was collected on level of eye fatigue, perception of job content, intervention usability, and musculoskeletal discomfort. Data was also collected on workspace level of illumination and posture during standardized tasks (assessed using RULA). Comparing baseline data to follow-up data for the intervention group, the use of the adjustable, LED task lights provided statistically significant, positive impacts on users' rating of discomfort, eye fatigue, perception of job content, and posture between baseline and the short-term follow up. Significant benefits to musculoskeletal comfort, posture, and visual comfort were documented when participants used the adjustable task lights. Participants' assessments of the light's usability, usefulness and desirability were positive. There were no negative results found with adjustable task light use.


HIV knowledge and health-seeking behavior in Zambézia Province, Mozambique.
HIV prevalence rates in Zambézia Province were estimated to be 12.6% in 2009. A number of educational campaigns have been aimed at improving HIV transmission and prevention knowledge among community members in an effort to reduce infection rates. These campaigns have also encouraged people to seek health care at clinical sites, instead of employing traditional healers to cure serious illness. The impact of these programs on the rural population has not been well documented. To assess the level of knowledge about HIV transmission and prevention and health-seeking behavior, we interviewed 349 people in 2009 using free response and multiple choice questionnaires. Over half reported first seeking treatment at a government health clinic; however, the majority of participants had visited a traditional healer in the past. Knowledge regarding prevention and transmission of HIV was primarily limited to the sexual origins of infection and the protective advantages of condom use. Increased educational level and having learned about HIV from a community health worker were associated with higher HIV prevention and transmission knowledge. Traditional healers and community health-care workers were both conduits of health information to our study participants. HIV education and use of clinical services may be facilitated by partnering more closely with these groups.


Elucidation of the disulfide-folding pathway of hirudin by a topology-based approach.
A theoretical model for the folding of proteins containing disulfide bonds is introduced. The model exploits the knowledge of the native state to favor the progressive establishment of native interactions. At variance with traditional approaches based on native topology, not all native bonds are treated in the same way; in particular, a suitable energy term is introduced to account for the special strength of disulfide bonds, as well as their ability to undergo intramolecular reshuffling. The model thus possesses the minimal ingredients necessary to investigate the much debated issue of whether the refolding process occurs through partially structured intermediates with native or non-native disulfide bonds. This strategy is applied to a context of particular interest, the refolding process of hirudin, a thrombin-specific protease inhibitor, for which conflicting folding pathways have been proposed. We show that the only two parameters in the model (temperature and disulfide strength) can be tuned to reproduce well a set of experimental transitions between species with different number of formed disulfides. This model is then used to provide a characterization of the folding process and a detailed description of the species involved in the rate-limiting step of hirudin refolding.


Treatment of an Ulnar Fracture Bone Defect Using Heterotopic Bone Autograft: A Case Report.
Autologous bone grafting has wide applications for the treatment of bony defects. Generally, cancellous or corticocancellous bone grafts are used depending on the characteristics and size of the bony defect and wound bed. The use of heterotopic bone as a potential source of bone graft has not been widely reported. We present a 56-year-old right-hand-dominant male victim of dog mauling who sustained a right ulnar fracture with a 5-cm bony defect, treated with the use of heterotopic bone autograft. Heterotopic bone can be successfully used as an autograft in the treatment of bony defects.


Mortality from intentional and unintentional injury among infants of young mothers in Colorado, 1986 to 1992.
To investigate the association between maternal age and other risk factors and infant injury deaths in the state of Colorado from 1986 to 1992. A retrospective cohort design was used to compare rates of unintentional and intentional infant injury mortality by maternal age group. A case-control design explored the importance of various risk factors, particularly maternal age, using multivariate logistic regression. The 2 case groups comprised all unintentional and intentional injury deaths in the first year of life. The control group was a random sample of both survivors and noninjury deaths selected from the entire birth cohort. The infant injury mortality rate for the 322766 live births in Colorado from 1986 to 1992 was 3.1 per 10000. Intentional injury death rates were highest for infants of teenaged mothers, peaking at 10.5 per 10000 live births for mothers aged 16 years. Unintentional injury death rates were highest for infants of mothers aged 20 to 24 years, peaking at 3.7 per 10000 live births for 22-year-old mothers. For intentional injury death, maternal marital status had a significant impact on maternal age; compared with the baseline group of married mothers older than 24 years, significantly higher risks were observed for infants of teenagers who were married (odds ratio [OR] = 32.0; 95% confidence interval[CI], 9.9-104.0) but also in infants of older mothers who were unmarried (OR = 3.6; 95% CI, 1.0-13.0 for unmarried mothers aged 20-24 years and (OR = 7.7; 95% CI, 2.4-25.0 for those > 24 years). Black race (OR = 3.5; 95% CI, 1.4-9.4) was also associated with intentional injury death. For unintentional injury death, the highest risk was for infants of mothers aged 20 to 24 years and unmarried (OR = 3.9; 95% CI, 1.7-9.3). Risk was also elevated for infants of married teenaged mothers (OR = 3.5; 95% CI, 0.7-17.8) but was not significantly different from the baseline group for unmarried teenagers, married 20- to 24-year-old mothers, or unmarried mothers aged 25 years or older. Risk was increased by the presence of older siblings (OR = 1.5 per sibling; 95% CI, 1.2-2.0). Maternal age and marital status significantly affect the rate of both unintentional and intentional infant injury mortality. These results suggest that child abuse prevention strategies should be targeted to teenaged mothers, and that strategies designed to prevent unintentional injuries should focus particularly on parents or caretakers of infants born to unmarried mothers in their early 20s as well as married teenagers.


Improving time to palliative care review with predictive modeling in an inpatient adult population: study protocol for a stepped-wedge, pragmatic randomized controlled trial.
Palliative care is a medical specialty centered on improving the quality of life (QOL) of patients with complex or life-threatening illnesses. The need for palliative care is increasing and with that the rigorous testing of triage tools that can be used quickly and reliably to identify patients that may benefit from palliative care. To that aim, we will conduct a two-armed stepped-wedge cluster randomized trial rolled out to two inpatient hospitals to evaluate whether a machine learning algorithm accurately identifies patients who may benefit from a comprehensive review by a palliative care specialist and decreases time to receiving a palliative care consult in hospital. This is a single-center study which will be conducted from August 2019 to November 2020 at Saint Mary's Hospital & Methodist Hospital both within Mayo Clinic Rochester in Minnesota. Clusters will be nursing units which will be chosen to be a mix of complex patients from Cardiology, Critical Care, and Oncology and had previously established relationships with palliative medicine. The stepped wedge design will have 12 units allocated to a design matrix of 5 treatment wedges. Each wedge will last 75 days resulting in a study period of 12 months of recruitment unless otherwise specified. Data will be analyzed with Bayesian hierarchical models with credible intervals denoting statistical significance. This intervention offers a pragmatic approach to delivering specialty palliative care to hospital patients in need using machine learning, thereby leading to high value care and improved outcomes. It is not enough for AI to be utilized by simply publishing research showing predictive performance; clinical trials demonstrating better outcomes are critically needed. Furthermore, the deployment of an AI algorithm is a complex process that requires multiple teams with varying skill sets. To evaluate a deployed AI, a pragmatic clinical trial can accommodate the difficulties of clinical practice while retaining scientific rigor. ClinicalTrials.gov NCT03976297 . Registered on 6 June 2019, prior to trial start.


Cerebral aneurysm regrowth and coil unraveling after incomplete Guglielmi detachable coil embolization: serial angiographical and histological findings.
A 69-year-old woman suffered subarachnoid hemorrhage due to rupture of an aneurysm at the tip of the basilar artery. The aneurysm was treated by endovascular treatment using Guglielmi detachable coils but the aneurysm dome could not be completely packed. Serial angiography at 10 and 18 months after embolization showed progressive regrowth of the aneurysm with loosening and unraveling of the packed coils. The patient died accidentally after head injury and autopsy was performed. Examination of the aneurysm showed further regrowth and "relative coil compaction" of the coil mass. Histological examination of the resected aneurysm showed no endothelialized membrane in the orifice and only minimal organized thrombus in the body of the aneurysm.


Racial and Sociodemographic Disparities in Internet Access and eHealth Intervention Utilization Among Veteran Smokers.
Access to the internet at home may be an important barrier to electronic health (eHealth) smoking cessation interventions. The current study explored possible sociodemographic disparities in access to the internet at home among veteran smokers. Data from participants proactively recruited and enrolled in a randomized smoking cessation effectiveness trial (N = 408) that compared a web-based smoking cessation intervention to Veterans Affairs (VA) usual care were used to examine the demographic attributes of smokers with and without internet access at home. Multivariable logistic regression was used to examine associations between demographic factors and home internet access. Data from patients randomized to the internet arm of the study (N = 205) were used to ascertain correlates of utilization of the intervention website. While the majority of the sample (82 %) endorsed access to the internet at home, veterans who were African-American, older, and not married were significantly less likely to have home internet access. Veterans who were African-American, older, less educated, had longer travel times to the nearest VA facility, and increased nicotine dependence were less likely to access the internet on a daily basis. While several sociodemographic variables (e.g., age, race, education, employment) were related to utilization of a free membership to a commercial, web-based smoking cessation intervention in bivariate analyses, only access to the internet at home was related to use of the smoking cessation site in adjusted results. These results highlight gaps in internet access and use among veterans and additionally underscore the importance of improving accessibility of eHealth interventions for low-income, minority, and socially disadvantaged veteran populations.


Distal Tibiofibular Syndesmotic Widening in Progressive Collapsing Foot Deformity.
Lateral overload in progressive collapsing foot deformity (PCFD) takes place as hindfoot valgus, peritalar subluxation, and valgus instability of the ankle increase. Fibular strain due to chronic lateral impingement may lead to distraction forces over the distal tibiofibular syndesmosis (DTFS). This study aimed to assess and correlate the severity of the foot and ankle offset (FAO) as a marker of progressive PCFD with the amount of DTFS widening and to compare it to controls. In this case-control study, 62 symptomatic patients with PCFD and 29 controls who underwent standing weightbearing computed tomography (WBCT) examination were included. Two fellowship-trained blinded orthopedic foot and ankle surgeons performed FAO (%) and DTFS area measurements (mm2). DTFS was assessed semiautomatically on axial-plane WBCT images, 1 cm proximal to the apex of the tibial plafond. Values were compared between patients with PCFD and controls, and Spearman correlation between FAO and DTFS area measurements was assessed. P values of less than .05 were considered significant. Patients with PCFD demonstrated significantly increased FAO and DTFS measurements in comparison to controls. A mean difference of 6.9% (P < .001) in FAO and 10.4 mm2 (P = .026) in DTFS was observed. A significant but weak correlation was identified between the variables, with a Þ of 0.22 (P = .03). A partition predictive model demonstrated that DTFS area measurements were highest when FAO values were between 7% and 9.3%, with mean (SD) values of 92.7 (22.4) mm2. To our knowledge, this was the first study to assess syndesmotic widening in patients with PCFD. We found patients with PCFD to demonstrate increased DTFS area measurements compared to controls, with a mean difference of approximately 10 mm2. A significantly weak positive correlation was found between FAO and DTFS area measurements, with the highest syndesmotic widening occurring when FAO values were between 7% and 9.3%. Our study findings suggest that chronic lateral impingement in patients with PCFD can result in a negative biomechanical impact on syndesmotic alignment, with increased DTFS stress and subsequent widening. Level III, retrospective comparative study.


Behavioral hearing thresholds between 0.125 and 20 kHz using depth-compensated ear simulator calibration.
The purpose of this study was to obtain behavioral hearing thresholds for frequencies between 0.125 and 20 kHz from a large population between 10 and 65 yr old using a clinically feasible calibration method expected to compensate well for variations in the distance between the eardrum and an insert-type sound source. Previous reports of hearing thresholds in the extended high frequencies (>8 kHz) have either used calibration techniques known to be inaccurate or specialized equipment not suitable for clinical use. Hearing thresholds were measured from 352 human subjects between 10 and 65 yr old having clinically normal-hearing thresholds (<20 dB HL) up to 4 kHz. An otoacoustic emission probe fitted with custom sound sources was used, and the stimulus levels individually tailored on the basis of an estimate of the insertion depth of the measurement probe. The calibrated stimulus levels were determined on the basis of measurements made at various depths of insertion in a standard ear simulator. Threshold values were obtained for 21 frequencies between 0.125 and 20 kHz using a modified Békésy technique. Forty-six of the subjects returned for a second measurement months later from the initial evaluation. In agreement with previous reports, hearing thresholds at extended high frequencies were found to be sensitive to age-related changes in auditory function. In contrast with previous reports, no gender differences were found in average hearing thresholds at most evaluated frequencies. Two aging processes, one faster than the other in time scale, seem to influence hearing thresholds in different frequency ranges. The standard deviation (SD) of test-retest threshold difference for all evaluated frequencies was 5 to 10 dB, comparable to that reported in the literature for similar measurement techniques but smaller than that observed for data obtained using the standard clinical procedure. The depth-compensated ear simulator-based calibration method and the modified Békésy technique allow reliable measurement of hearing thresholds over the entire frequency range of human hearing. Hearing thresholds at the extended high frequencies are sensitive to aging and reveal subtle differences, which are not evident in the frequency range evaluated regularly (≤8 kHz). Previously reported gender-related differences in hearing thresholds may be related to ear-canal acoustics and the calibration procedure and not because of differences in hearing sensitivity.


Immunotherapy in urothelial cancer, part 1: T-cell checkpoint inhibition in advanced or metastatic disease.
Cancer of the urothelium is the sixth most common cancer in the United States and is seen predominantly in men. Most cases of this disease present as non-muscle-invasive bladder cancer (NMIBC), with cancer recurrence or progression to muscle-invasive cancer in more than 50% of patients after initial therapy. NMIBC is an immune-responsive disease, as indicated by the use of intravesical bacillus Calmette-Guérin as treatment for more than 3 decades. More recently, immunotherapy has seen much progress in a variety of cancers, including advanced and metastatic bladder cancer, in which historical 5-year survival rates are approximately 15%. The advent of T-cell checkpoint inhibitors, especially those directed at programmed death 1 (PD-1) and its ligand (PD-L1), has had a significant effect on the therapy of advanced urothelial cancer. This had led to accelerated approval by the US Food and Drug Administration for atezolizumab and nivolumab in advanced urothelial cancer previously treated with platinum-based chemotherapy. In addition, level 1 evidence supports the use of pembrolizumab over single-agent tubulin-directed chemotherapy in the same setting. Several other treatments with immune-mediating mechanisms of action are in development and hold great promise, including monoclonal antibodies directed at other checkpoint molecules, oncolytic virus therapy, adoptive T-cell therapy, combination immunotherapy, and antibody-drug conjugates. This review focuses on the recent development of T-cell checkpoint inhibitors in advanced and metastatic urothelial cancer and addresses their potential use in combination. It also discusses a spectrum of novel immunotherapies with potential use in urothelial cancer.


Advancing methods for research on household water insecurity: Studying entitlements and capabilities, socio-cultural dynamics, and political processes, institutions and governance.
Household water insecurity has serious implications for the health, livelihoods and wellbeing of people around the world. Existing methods to assess the state of household water insecurity focus largely on water quality, quantity or adequacy, source or reliability, and affordability. These methods have significant advantages in terms of their simplicity and comparability, but are widely recognized to oversimplify and underestimate the global burden of household water insecurity. In contrast, a broader definition of household water insecurity should include entitlements and human capabilities, sociocultural dynamics, and political institutions and processes. This paper proposes a mix of qualitative and quantitative methods that can be widely adopted across cultural, geographic, and demographic contexts to assess hard-to-measure dimensions of household water insecurity. In doing so, it critically evaluates existing methods for assessing household water insecurity and suggests ways in which methodological innovations advance a broader definition of household water insecurity.


Azetidine-induced accumulation of class I small heat shock proteins in the soluble fraction provides thermotolerance in soybean seedlings.
Accumulation of class I small heat shock proteins (sHSPs) is induced by the proline analog, azetidine-2-carboxylic acid (Aze) in soybean seedlings to a level similar to that induced by exposure to 40 degrees C. However, only the treatment with 10 mM Aze for 6 h and subsequently with 10 mM proline for 24 h protected the seedlings from damage during subsequent exposure to 45 degrees C as assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. A chaperone activity assay showed that the purified class I sHSPs induced by Aze were functional in vitro and protected proteins from thermal denaturation. Amino acid composition analysis indicated that Aze was not incorporated into de novo synthesized class I sHSPs. Accumulation of class I sHSPs in the soluble post-ribosomal supernatant fraction was found to be important for acquisition of thermotolerance. We suggest that both the accumulation of class I sHSPs and their presence in the soluble fraction are important for establishment of thermotolerance.


Human impact on genetic diversity of Toxoplasma gondii: example of the anthropized environment from French Guiana.
In French Guiana, severe cases of toxoplasmosis in immunocompetent patients are associated with atypical strains of Toxoplasma gondii linked to a wild neotropical rainforest cycle and a higher genetic diversity than usually observed for T. gondii isolates from anthropized environment. This raises the question of the impact of anthropization of the natural environment, on genetic diversity and on the population structure of T. gondii. However, few data are available on strains circulating in the anthropized areas from French Guiana. Seropositive animals originating mainly from anthropized sub-urban areas and punctually from wild environment in French Guiana were analyzed for T. gondii isolation and genotyping. Thirty-three strains were obtained by bioassay in mice and compared with 18 previously reported isolates chiefly originating from the Amazon rainforest. The genotyping analysis performed with 15 microsatellite markers located on 12 different chromosomes revealed a lower genetic diversity in the anthropized environment. Results were analyzed in terms of population structure by clustering methods, Neighbor-joining trees reconstruction based on genetic distances, F(ST,) Mantel's tests and linkage disequilibrium. They clearly showed a genetic differentiation between strains associated to the anthropized environment and those associated to the wild, but with some inbreeding between them. The majority of strains from the anthropized environment were clustered into additional lineages of T. gondii that are common in the Caribbean. In conclusion the two environmental populations "wild" and "anthropized" were genetically well differentiated. The anthropization of the environment seems to be accompanied with a decreased diversity of T. gondii associated with a greater structure of the populations. We detected potential interpenetration and genetic exchanges between these two environmental populations. As a higher pathogenicity in human of "wild" genotypes has been described, the interpenetration of both environments leads to hybridization between strains that may be at risk for human health.


Postoperative Recurrence of Instability Due to New Anterior Glenoid Rim Fractures After Arthroscopic Bankart Repair.
Computed tomography (CT) sometimes reveals a new fracture of the anterior glenoid rim in patients with postoperative recurrence of instability after arthroscopic Bankart repair using suture anchors, but there have been few previous reports about such fractures. The placement of a large number of suture anchors during arthroscopic Bankart repair might be associated with a new glenoid rim fracture. Cohort study; Level of evidence, 3. Screw-in metal suture anchors were used until June 2011 and suture-based soft anchors from July 2011. A follow-up of at least 2 years was conducted for 128 shoulders treated using metal anchors (metal anchor group) and 129 shoulders treated using soft anchors (soft anchor group). The frequency and features of new glenoid rim fractures were investigated, and the influence of the number of suture anchors and other factors on fractures was also assessed. There were 19 shoulders (14.8%) with postoperative recurrence in the metal anchor group and 23 shoulders (17.8%) in the soft anchor group. Among 37 shoulders evaluated by CT at recurrence, a new glenoid rim fracture was detected in 13 shoulders (35.1%; 5 shoulders in the metal anchor group and 8 shoulders in the soft anchor group). A fracture at the anchor insertion site was recognized in 4 shoulders from the metal anchor group and 6 shoulders from the soft anchor group, although linear fractures connecting several anchor holes were only seen in the soft anchor group. While new glenoid fractures occurred regardless of the number of suture anchors used, new fractures were significantly more frequent in teenagers at surgery and in junior high school or high school athletes. Such fractures did not only occur in contact athletes but were also found in overhead athletes. Postoperative recurrence of instability associated with a new glenoid rim fracture along the suture anchor insertion site was frequent after arthroscopic Bankart repair. These fractures might be related to placing multiple soft suture anchors in a linear arrangement.


Copy-number variation and protein expression of DOT1L in pancreatic adenocarcinoma as a potential drug target.
Adenocarcinoma of the pancreas has a poor prognosis. At present, no relevant personalized targets have been identified. Sequencing studies have implicated gene alterations of disruptor of telomeric silencing 1 like histone lysine methyltransferase (DOT1L) in pancreatic adenocarcinoma. DOT1L is part of the histone modification system and catalyzes methylation of H3K79, which is crucial in cell signaling and DNA damage repair. DOT1L is considered to be a target of therapy in mixed lineage leukemia gene-deficient leukemia cases and a potential target in breast carcinoma. The frequencies and importance of DOT1L copy-number variations and their specific correlation with protein expression in adenocarcinoma of the pancreas have yet to be investigated. In the present study, tissue microarrays of 230 resected pancreatic adenocarcinoma cases were constructed. The tumor tissue was analyzed using fluorescence in situ hybridization (FISH) and immunohistochemistry. In total, 10/225 carcinoma cases (4.4%) analyzed by immunohistochemistry demonstrated intense nuclear protein expression of DOT1L and in 9/224 tumors analyzed using FISH (4.0%), copy-number variations (CNV) were detectable. No DOT1L amplification was detected in the carcinoma cohort. To the best of our knowledge, the present study describes for the first time the frequency of CNV of DOT1L using the gold standard fluorescence in situ hybridization (FISH) and their specific correlation to the protein expression in adenocarcinomas of the pancreas. Although the positive cases by immunohistochemistry and copy-number variations by FISH were not congruent with each other, the data suggest a potential role for DOT1L in a small subset of pancreatic cancer cases. The significance of the two analysis methods concerning their druggability in pancreatic adenocarcinoma requires further studies.


Ethical issues in artificial nutrition and hydration.
From the time of Hippocrates, approximately 2500 years ago, medical ethics has been seen as an essential complement to medical science in pursuit of the healing art of medicine. This is no less true today, not only for physicians but also for other essential professionals involved in patient care, including clinical nutrition support practitioners. One aspect of medical ethics that the clinical nutritionist must face involves decisions to provide, withhold, or withdraw artificial nutrition and hydration. Such a decision is not only technical but often has a strong moral component as well. Although it is the physician who writes any such order, the clinical nutritionist as fellow professional should be a part not only of the scientific aspects of the order but of the moral discourse leading to such an order and may certainly be involved in counseling physicians, other healthcare providers, patients, and families alike. This paper is intended to give the clinical nutritionist a familiarity with the discipline of medical ethics and its proper relationship to medical science, politics, and law. This review will then offer a more specific analysis of the ethical aspects of decisions to initiate, withhold, or withdraw artificial nutrition and hydration (ANH) and offer particular commentary on the ethically significant pronouncements of Pope John Paul II in March of 2004 related to vegetative patients and artificial or "assisted" nutrition and hydration.


Arbitrarily large heteroclinic networks in fixed low-dimensional state space.
We consider heteroclinic networks between n∈N nodes where the only connections are those linking each node to its two subsequent neighboring ones. Using a construction method where all nodes are placed in a single one-dimensional space and the connections lie in coordinate planes, we show that it is possible to robustly realize these networks in R6 for any number of nodes n using a polynomial vector field. This bound on the space dimension (while the number of nodes in the network goes to ∞) is a novel phenomenon and a step toward more efficient realization methods for given connection structures in terms of the required number of space dimensions. We briefly discuss some stability properties of the generated heteroclinic objects.


Pelvic floor muscle examination in female chronic pelvic pain.
To determine if women with self-reported chronic pelvic pain (CPP) were more likely to have positive findings on two vaginal pelvic floor muscle (PFM) tests compared to women without CPP when the examiner was blinded to pain status. This was a prospective, cross-sectional study. Blinded examiners performed two vaginal pelvic floor tests (tenderness and strength) on 48 participants: 19 with self-reported CPP and 29 who were pain-free. Relative frequency of positive findings between groups and the total number of positive physical examination findings were calculated. Women with self-reported CPP were more likely to have PFM tenderness (63.2% with physician [M.D.] examiners [board certified in physical medicine and rehabilitation] and 73.7% with physical therapist [P.T.] examiners) as compared to pain-free participants (Fisher's exact test [FET]), 48 p < 0.001 with M.D., p < 0.001 with P.T.). PFM weakness was not more likely in women with CPP (31.6% with M.D., 42.1% with P.T.) as compared with pain-free participants (48.3% with M.D., 17.2% with P.T.) (FET, 48 p = 0.37 with M.D., p = 0.096 with P.T.). PFM tenderness is found more frequently in women with self-reported CPP than in pain-free women. PFM strength did not differentiate CPP from pain-free participants. Improved standardization of the PFM examination across disciplines may be helpful in distinguishing subgroups and treating women with CPP.


Psychometric evaluation of a cultural competency assessment instrument for health professionals.
Few valid and reliable measures exist for health care professionals interested in determining their levels of cultural and linguistic competence. To evaluate the measurement properties of the Cultural Competence Health Practitioner Assessment (CCHPA-129). The CCHPA-129 is a 129-item web-based instrument, developed by the National Center for Cultural Competence. Responses on the CCHPA -129 were examined using factor analysis; Rasch modeling; and differential item functioning across race, ethnicity, sex, and profession. A total of 2504 practitioners, including 1864 nurses (RN/LPN/BSN); 341 clinicians (PA/NP); and 299 physicians (MD/DO), who completed the CCHPA-129 online between 2005 and 2008. Three factors representing domains of Knowledge, Adapting Practice, and Promoting Health for culturally and linguistically diverse populations accounted for 46% of the variance. Among Knowledge factor items, 53% (23/43) fit the Rasch model, item difficulties ranged from -1.01 logits (least difficult) to +1.11 logits (most difficult), separation index (SI) 13.82, and Cronbach's α 0.92. Forty-seven percent (21/44) Adapting Practice factor items fit the model, item difficulties -0.07 to +1.11 logits, SI 11.59, Cronbach's α 0.88; and 58% (23/39). Promoting Health factor items fit the model, item difficulties -1.01 to +1.38 logits, SI 22.64, Cronbach's α 0.92. Early evidence of validity was established by known groups having statistically different scores. The 67-item CCHPA-67 is psychometrically sound. This shorted instrument can be used to establish associations between practitioners' cultural and linguistic competence and health outcomes as well as to evaluate interventions to increase practitioners' cultural and linguistic competence.


WC1 is a hybrid γδ TCR coreceptor and pattern recognition receptor for pathogenic bacteria.
WC1 proteins are uniquely expressed on γδ T cells and belong to the scavenger receptor cysteine-rich (SRCR) superfamily. While present in variable, and sometimes high, numbers in the genomes of mammals and birds, in cattle there are 13 distinct genes (WC1-1 to WC1-13). All bovine WC1 proteins can serve as coreceptors for the TCR in a tyrosine phosphorylation dependent manner, and some are required for the γδ T cell response to Leptospira. We hypothesized that individual WC1 receptors encode Ag specificity via coligation of bacteria with the γδ TCR. SRCR domain binding was directly correlated with γδ T cell response, as WC1-3 SRCR domains from Leptospira-responsive cells, but not WC1-4 SRCR domains from Leptospira-nonresponsive cells, bound to multiple serovars of two Leptospira species, L. borgpetersenii, and L. interrogans. Three to five of eleven WC1-3 SRCR domains, but none of the eleven WC1-4 SRCR domains, interacted with Leptospira spp. and Borrelia burgdorferi, but not with Escherichia coli or Staphylococcus aureus. Mutational analysis indicated that the active site for bacterial binding in one of the SRCR domains is composed of amino acids in three discontinuous regions. Recombinant WC1 SRCR domains with the ability to bind leptospires inhibited Leptospira growth. Our data suggest that WC1 gene arrays play a multifaceted role in the γδ T cell response to bacteria, including acting as hybrid pattern recognition receptors and TCR coreceptors, and they may function as antimicrobials.


Genetics of methyl-accepting chemotaxis proteins in Escherichia coli: organization of the tar region.
The tar locus of Escherichia coli specifies one of the major species of methyl-accepting proteins involved in the chemotactic behavior of this organism. The physical and genetic organization of the tar region was investigated with a series of specialized lambda transducing phages and plasmid clones. The tar gene was mapped at the promoter-proximal end of an operon containing five other chemotaxis-related loci. Four of those genes (cheR, cheB, cheY and cheZ) are required for all chemotactic responses; consequently, polar mutations in the tar gene resulted in a generally nonchemotactic phenotype. The fifth gene, tap, was mapped between the tar and cheR loci and specified the production of a 65-kilodalton methyl-accepting protein. Unlike the tar locus, which is required for chemotaxis to aspartate and maltose, mutants lacking only the tap function had no obvious defects in chemotactic ability. Genetic and physical maps of the tar-tap region were constructed with Mu d1 (Apr lac) insertion mutations, whose polar properties conferred a phenotype suitable for deletion mapping studies. Restriction endonuclease analyses of phage and plasmid clones indicated that all of the genetic coding capacity in the tar region is now accounted for.