Functional analysis of human T cell subsets defined by monoclonal antibodies. I. Collaborative T-T interactions in the immunoregulation of B cell differentiation.
T-B and T-T interactions involved in the regulation of PWM-triggered human B cell differentiation were studied in vitro. Functionally distinct human T cell subsets were isolated by C-mediated lysis by using the monoclonal antibodies OKT4 and OKT8. Graded numbers of either untreated or irradiated T cell subsets were added to autologous B cells, and total antibody synthesis was measured after 5 to 6 days of culture by using a highly sensitive reverse hemolytic plaque assay. The data indicate that a) the helper activity that is exclusively contained within the OKT4+ population is radiosensitive. Only at high T/B ratios can this radiosensitivity be overcome; b) the OKT8+ population contains radiosensitive cells important in suppressing B cell differentiation, and c) the suppression induced with OKT8+ cells requires the presence of radiosensitive OKT4+ cells. Thus, OKT8+ cells added to cultures containing B cells and irradiated OKT4+ cells do not suppress the PFC response. Addition of unirradiated OKT4+ cells to these cultures permits reexpression of suppression by OKT8+ cells. It is concluded that two radiosensitive cells, one within the OKT4+ population and the other within the OKT8+ population, collaborate to induce suppression. Possible mechanisms for this suppressive interaction including induction of suppressor precursor cells within the OKT4+ population or inhibition of OKT4+ helper cells by OKT8+ cells are discussed.


Persistence of anticancer activity in berry extracts after simulated gastrointestinal digestion and colonic fermentation.
Fruit and vegetable consumption is associated at the population level with a protective effect against colorectal cancer. Phenolic compounds, especially abundant in berries, are of interest due to their putative anticancer activity. After consumption, however, phenolic compounds are subject to digestive conditions within the gastrointestinal tract that alter their structures and potentially their function. However, the majority of phenolic compounds are not efficiently absorbed in the small intestine and a substantial portion pass into the colon. We characterized berry extracts (raspberries, strawberries, blackcurrants) produced by in vitro-simulated upper intestinal tract digestion and subsequent fecal fermentation. These extracts and selected individual colonic metabolites were then evaluated for their putative anticancer activities using in vitro models of colorectal cancer, representing the key stages of initiation, promotion and invasion. Over a physiologically-relevant dose range (0-50 µg/ml gallic acid equivalents), the digested and fermented extracts demonstrated significant anti-genotoxic, anti-mutagenic and anti-invasive activity on colonocytes. This work indicates that phenolic compounds from berries undergo considerable structural modifications during their passage through the gastrointestinal tract but their breakdown products and metabolites retain biological activity and can modulate cellular processes associated with colon cancer.


Thermal imaging as a tool for evaluating tumor treatment efficacy.
Breast cancer is the most frequently diagnosed cancer among women in the Western world. Thermography is a nonionizing, noninvasive, portable, and low-cost method that can be used in an outpatient clinic. It was tried as a tool to detect breast cancer tumors, however, it had too many false readings. Thermography has been extensively studied as a breast cancer detection tool but was not used as a treatment monitoring tool. The purpose of this study was to investigate the possibility of using thermal imaging as a feedback system to optimize radiation therapy. Patients were imaged with a thermal camera prior and throughout the radiotherapy sessions. At the end of the session, the images were analyzed for temporal vasculature changes through vessels segmentation image processing tools. Tumors that were not responsive to treatment were observed before the radiation therapy sessions were concluded. Assessing the efficacy of radiotherapy during treatment makes it possible to change the treatment regimen, dose, and radiation field during treatment as well as to individualize treatment schedules to optimize treatment effectiveness.


Dynamical systems for discovering protein complexes and functional modules from biological networks.
Recent advances in high throughput experiments and annotations via published literature have provided a wealth of interaction maps of several biomolecular networks, including metabolic, protein-protein, and protein-DNA interaction networks. The architecture of these molecular networks reveals important principles of cellular organization and molecular functions. Analyzing such networks, i.e., discovering dense regions in the network, is an important way to identify protein complexes and functional modules. This task has been formulated as the problem of finding heavy subgraphs, the Heaviest k-Subgraph Problem (k-HSP), which itself is NP-hard. However, any method based on the k-HSP requires the parameter k and an exact solution of k-HSP may still end up as a "spurious" heavy subgraph, thus reducing its practicability in analyzing large scale biological networks. We proposed a new formulation, called the rank-HSP, and two dynamical systems to approximate its results. In addition, a novel metric, called the Standard deviation and Mean Ratio (SMR), is proposed for use in "spurious" heavy subgraphs to automate the discovery by setting a fixed threshold. Empirical results on both the simulated graphs and biological networks have demonstrated the efficiency and effectiveness of our proposal.


Cherry: mobile application for children with cancer.
The Cherry project seeks to address the information needs of young cancer patients, their parents, and health care providers. It aims at helping the patients to understand various aspects of their disease and treatment, and allow them to assess and record their disease related quality of life. It uses elements of social media to offer a meeting point with the physician and peers. Information is presented in a way that is both understandable and appealing to young children in school age and adolescents. Preschool children will be studied as a separate user group to address their needs and possibilities to meet them. The Cherry system wants to utilize Internet and mobile technologies to benefit patient outcome.


Entrustable Professional Activities for Entering Residency: Establishing Common Osteopathic Performance Standards in the Transition From Medical School to Residency.
Entrustable professional activities (EPAs) are measurable units of observable professional practice that can be entrusted to an unsupervised trainee. They were first introduced as a method of operationalizing competency-based medical education in graduate medical education. The American Association of Medical Colleges subsequently used EPAs to establish the core skills that medical students must be able to perform before they enter residency training. A recently published guide provides descriptions, guidelines, and rationale for implementing and assessing the core EPAs from an osteopathic approach. These osteopathically informed EPAs can allow schools to more appropriately assess a learner's whole-person approach to a patient, in alignment with the philosophy of the profession. As the single accreditation system for graduate medical education moves forward, it will be critical to integrate EPAs into osteopathic medical education to demonstrate entrustment of medical school graduates. The authors describe the collaborative process used to establish the osteopathic considerations added to EPAs and explores the challenges and opportunities for undergraduate osteopathic medical education.


Frailty Checkup Supporters' Intentions to Participate in Human-Resource Development and Training Activities.
Frailty prevention is one of social prescriptions for an aging society. That requires community level intervention. This study examined frailty checkup supporters' (FCSs') intentions to engage in human-resource development and training activities (i.e., leadership activities) and related factors. Cross-sectional study. Three municipalities in suburban area, Eastern Japan. Forty-five of 59 FCSs completed anonymous self-administered questionnaires. Questionnaire sought information regarding their sociodemographic data, their perceptions and experiences of FCS activities, and their intentions to participate in human-resource development and training activities. Participants were divided into a high intention (HI) and low intention group (LI). The two groups were compared using quantitative and qualitative data. Eleven FCSs reported intending to engage in leader-related activities. Factors associated with FCSs' intentions were finding FCS activities rewarding and the willingness to continue performing FCS activities. The participants who changed their daily activities (p = .041) and perceptions regarding contributing to the community (p = .018) showed significantly higher intention than LI participants. Free description about the changes in perceptions and lifestyles as a consequence of participating in FCS activities were analyzed qualitatively. FCSs who changed their daily activities and perspectives about contributing to the community described positive changes in both the groups. Meanwhile, FCSs who did not change their daily activities and perspectives about contributing to the community described their reason only in the LI group. The results might encourage FCSs to participate in training and guidance activities, as they have positive experiences and receive recognition obtained through participation in such activities.


The transporter-like protein inebriated mediates hyperosmotic stimuli through intracellular signaling.
We cloned the inebriated homologue MasIne from Manduca sexta and expressed it in Xenopus laevis oocytes. MasIne is homologous to neurotransmitter transporters but no transport was observed with a number of putative substrates. Oocytes expressing MasIne respond to hyperosmotic stimulation by releasing intracellular Ca(2+), as revealed by activation of the endogenous Ca(2+)-activated Cl(-) current. This Ca(2+) release requires the N-terminal 108 amino acid residues of MasIne and occurs via the inositol trisphosphate pathway. Fusion of the N terminus to the rat gamma-aminobutyric acid transporter (rGAT1) also renders rGAT1 responsive to hyperosmotic stimulation. Immunohistochemical analyses show that MasIne and Drosophila Ine have similar tissue distribution patterns, suggesting functional identity. Inebriated is expressed in tissues and cells actively involved in K(+) transport, which suggests that it may have a role in ion transport, particularly of K(+). We propose that stimulation of MasIne releases intracellular Ca(2+) in native tissues, activating Ca(2+)-dependent K(+) channels, and leading to K(+) transport.


Transcriptional regulation of PRPF31 gene expression by MSR1 repeat elements causes incomplete penetrance in retinitis pigmentosa.
PRPF31-associated retinitis pigmentosa presents a fascinating enigma: some mutation carriers are blind, while others are asymptomatic. We identify the major molecular cause of this incomplete penetrance through three cardinal features: (1) there is population variation in the number (3 or 4) of a minisatellite repeat element (MSR1) adjacent to the PRPF31 core promoter; (2) in vitro, 3-copies of the MSR1 element can repress gene transcription by 50 to 115-fold; (3) the higher-expressing 4-copy allele is not observed among symptomatic PRPF31 mutation carriers and correlates with the rate of asymptomatic carriers in different populations. Thus, a linked transcriptional modifier decreases PRPF31 gene expression that leads to haploinsufficiency. This result, taken with other identified risk alleles, allows precise genetic counseling for the first time. We also demonstrate that across the human genome, the presence of MSR1 repeats in the promoters or first introns of genes is associated with greater population variability in gene expression indicating that copy number variation of MSR1s is a generic controller of gene expression and promises to provide new insights into our understanding of gene expression regulation.


Obesogenic neighbourhoods: the impact of neighbourhood restaurants and convenience stores on adolescents' food consumption behaviours.
To examine the relationship between the neighbourhood food environment and dietary intake among adolescents. Cross-sectional design using: (i) a geographic information system to assess characteristics of the neighbourhood food environment and neighbourhood socio-economic status; (ii) the modified Healthy Eating Index (HEI) to assess participants' overall diet quality; and (iii) generalized linear models to examine associations between HEI and home and school food environmental correlates. Mid-sized Canadian city in Ontario, Canada. Participants Grade 7 and 8 students (n 810) at twenty-one elementary schools. Students living in neighbourhoods with a lower diversity of land-use types, compared with their higher diversity counterparts, had higher HEI scores (P < 0.05). Students with more than 1 km between their home and the nearest convenience store had higher HEI scores than those living within 1 km (P < 0.01). Students attending schools with a distance further than 1 km from the nearest convenience store (P < 0.01) and fast-food outlet (P < 0.05) had higher HEI scores than those within 1 km. Those attending schools with three or more fast-food outlets within 1 km had lower HEI scores than those attending schools with no fast-food outlet in the school surroundings (P < 0.05). Close proximity to convenience stores in adolescents' home environments is associated with low HEI scores. Within adolescents' school environments, close proximity to convenience and fast-food outlets and a high density of fast-food outlets are associated with low HEI scores.


Differential roles of STAT1alpha and STAT1beta in fludarabine-induced cell cycle arrest and apoptosis in human B cells.
Signal transducer and activator of transcription 1 (STAT1), a transcription factor known to participate in antiviral responses, acts as a tumor suppressor inhibiting cell growth and promoting apoptosis. To study the role of STAT1 in DNA damage-induced apoptosis in B lymphocytes, its active form, STAT1alpha, was specifically inhibited by the overexpression of STAT1beta, the STAT1alpha truncated inhibitory isoform. An episomal vector with a tetracycline-inducible bidirectional promoter was created to induce the expression of 2 proteins, STAT1beta and enhanced green fluorescence protein (EGFP). The same vector was used to overexpress STAT1alpha as a control. Expression of STAT1beta inhibited the phosphorylation, the DNA-binding activity, and the transcriptional activity of STAT1alpha, as well as the expression of STAT1alpha target genes such as p21WAF1/CIP1, TAP1, IRF1, and PKR. Inhibiting STAT1alpha by STAT1beta increased the growth rate of transfected cells and their resistance to fludarabine-induced apoptosis and cell cycle arrest. Overexpressing STAT1beta reversed the negative regulation of Mdm2 expression observed after treatment with interferon-gamma (IFN-gamma), which activates STAT1, or with fludarabine. Nuclear translocation of p53 after fludarabine treatment was decreased when STAT1beta was overexpressed, and it was increased when STAT1alpha was induced. Oligonucleotide pull-down experiments showed a physical STAT1/p53 interaction. Our results show that imbalance between the antiproliferative/proapoptotic isoform STAT1alpha and the proliferative isoform STAT1beta is likely to play a crucial role in the regulation of proliferation and apoptosis and that STAT1alpha may regulate p53 activity and sensitize B cells to fludarabine-induced apoptosis.


Association of adiponectin gene expression with atrial fibrillation in a Pakistani populace.
Adiponectin, an adipocytokine produced and secreted by adipose tissue, has anti-diabetic, anti-atherogenic, and anti-inflammatory properties. This case-control study was aimed to assess the expression and serum levels of adiponectin in subject suffereing from atrial fibrillation (AF). The study's subjects (n = 690) were enrolled from the Punjab Institute of Cardiology, Lahore and were grouped into control, AF without Metabolic syndrome (MetS), and AF with MetS groups. Along with the collection of demographic data, an analysis of adiponectin and biochemical parameters were performed. A highly significant difference in serum levels of adiponectin was observed among the control, AF without MetS, and AF with MetS groups (61.61 ± 45.30 ng/ml, 37.20 ± 19.46 ng/ml, 63.78 ± 61.69 ng/ml). The expression analysis of adiponectin was decreased (n-fold = ̴ 0.30) in AF without MetS group as compared to control group (n-fold = ~ 1.16) but increased in AF with MetS group (n-fold = ̴ 6.26). The correlation analysis revealed a highly significant positive relationship between the expression of the adiponectin gene with waist-to-hip ratio (WHR) in AF without MetS group. Whereas, serum adiponectin was negatively related to serum triglycerides (TG) in AF with MetS group. In multiple regression analysis using adiponectin expression as the dependent variable, WHR was a determinant in AF without MetS. Whereas, when serum adiponectin was used as the dependent variable, serum TG was the determinant in group AF with MetS. The present study implicates that decreased expression and serum levels of adiponectin were associated with the development of AF in which WHR and serum TG also contributed towards the onset of atrial fibrillation.


Binding of purified Bacillus sphaericus binary toxin and its deletion derivatives to Culex quinquefasciatus gut: elucidation of functional binding domains.
Highly larvicidal strains of Bacillus sphaericus produce a binary toxin composed of 51 and 42 kDa proteins which binds to sharply delineated regions of the gastric caecum and posterior midgut of susceptible larvae of the mosquito Culex quinquefasciatus. To investigate the role of the individual subunits and the organization of functional binding regions within the toxin, plasmids were constructed for the expression in Escherichia coli of the toxin proteins and their NH2- and COOH-terminal deletion derivatives as fusions with glutathione S-transferase (GST). Toxin proteins were purified by affinity chromatography followed by cleavage from the GST carrier with thrombin. The LC50 values for the purified toxin proteins and their deletion derivatives were determined. The binding patterns of fluorescently labelled toxin suggested that the 51 kDa protein is the primary binding component of the toxin and mediates the regional binding and internalization of the 42 kDa protein. Examination of the toxin deletion derivatives revealed that the NH2-terminal region of the 51 kDa protein was required for binding to the larval gut, whilst the COOH-terminal region was responsible for interacting with the 42 kDa protein. Toxicity was strongly correlated with the subsequent internalization of the toxin, probably by endocytosis.


Vertical guidance performance analysis of the L1-L5 dual-frequency GPS/WAAS user avionics sensor.
This paper investigates the potential vertical guidance performance of global positioning system (GPS)/wide area augmentation system (WAAS) user avionics sensor when the modernized GPS and Galileo are available. This paper will first investigate the airborne receiver code noise and multipath (CNMP) confidence (σair). The σair will be the dominant factor in the availability analysis of an L1-L5 dual-frequency GPS/WAAS user avionics sensor. This paper uses the MATLAB Algorithm Availability Simulation Tool (MAAST) to determine the required values for the σair, so that an L1-L5 dual-frequency GPS/WAAS user avionics sensor can meet the vertical guidance requirements of APproach with Vertical guidance (APV) II and CATegory (CAT) I over conterminous United States (CONUS). A modified MAAST that includes the Galileo satellite constellation is used to determine under what user configurations WAAS could be an APV II system or a CAT I system over CONUS. Furthermore, this paper examines the combinations of possible improvements in signal models and the addition of Galileo to determine if GPS/WAAS user avionics sensor could achieve 10 m Vertical Alert Limit (VAL) within the service volume. Finally, this paper presents the future vertical guidance performance of GPS user avionics sensor for the United States' WAAS, Japanese MTSAT-based satellite augmentation system (MSAS) and European geostationary navigation overlay service (EGNOS).


A practical approach to storage and retrieval of high-frequency physiological signals.
Storage of physiological waveform data for retrospective analysis presents significant challenges. Resultant data can be very large, and therefore becomes expensive to store and complicated to manage. Traditional database approaches are not appropriate for large scale storage of physiological waveforms. Our goal was to apply modern time series compression and indexing techniques to the problem of physiological waveform storage and retrieval. We deployed a vendor-agnostic data collection system and developed domain-specific compression approaches that allowed long term storage of physiological waveform data and other associated clinical and medical device data. The database (called AtriumDB) also facilitates rapid retrieval of retrospective data for high-performance computing and machine learning applications. A prototype system has been recording data in a 42-bed pediatric critical care unit at The Hospital for Sick Children in Toronto, Ontario since February 2016. As of December 2019, the database contains over 720,000 patient-hours of data collected from over 5300 patients, all with complete waveform capture. One year of full resolution physiological waveform storage from this 42-bed unit can be losslessly compressed and stored in less than 300 GB of disk space. Retrospective data can be delivered to analytical applications at a rate of up to 50 million time-value pairs per second. Stored data are not pre-processed or filtered. Having access to a large retrospective dataset with realistic artefacts lends itself to the process of anomaly discovery and understanding. Retrospective data can be replayed to simulate a realistic streaming data environment where analytical tools can be rapidly tested at scale.


Global health training in pediatric residency programs.
Our goal was to describe current resident interest, participation, curricula, resources, and obstacles related to global health training within pediatric residency programs. We conducted a cross-sectional survey of the 201 accredited pediatric residency programs in the United States, Puerto Rico, and the Caribbean from October 2006 to January 2007. Survey topics included resident interest and participation in electives, training opportunities, program support, and educational curricular content related to global health. Of the 201 surveyed pediatric residency programs, 106 (53%) responded. Fifteen percent of responding programs reported that a majority of their residents were interested in global health. Fifty-two percent offered a global health elective within the previous year, and 47% had formally incorporated global health into their training curricula. Six percent of the programs reported a formalized track or certificate in global health. The median number of residents per program participating in global health electives within the previous year was 0 during postgraduate year 1, 1 during postgraduate year 2, and 2 during postgraduate year 3. The median number of all residents per program participating in a global health elective in the previous year was 3 (7.4% of program size). Among programs that offered a global health elective, support to participating residents included prerequisite clinical training (36%), cultural orientation (36%), language training (15%), faculty mentorship (82%), and post-elective debriefing (77%). Fourteen percent of the programs provided full funding for resident electives. Characteristics of pediatric residency programs that were significantly associated with higher resident participation in a global health elective were larger program size, university affiliation, greater reported resident interest, and faculty involvement in global health. More than half of the pediatric residency programs surveyed offered a global health elective in the previous year. An American Academy of Pediatrics survey 10 years earlier had shown 1 of 4 programs with global health electives. Observance of American Academy of Pediatrics consensus guidelines for global health electives varied widely among programs, and additional efforts should focus on resident preparation, mentorship, and funding.


[Total spinal block after posterior lumbar plexus blockade: case report.].
Lumbar plexus blockade can be very useful in surgical procedures of the hip, thigh, and knee, but it should be performed by an experienced anesthesiologist due to potential complications. The current report presents a case of total spinal block after posterior lumbar plexus blockade and discusses the possible pathophysiological mechanisms. Male patient, 34 years old, 97 kg, physical status ASA I, scheduled for total hip arthroplasty. After general anesthesia, a right posterior lumbar plexus blockade was performed with the adjunct of a peripheral nerve stimulator. The needle was introduced to a depth of 8 cm, perpendicular to the skin, 4 cm from the mid line, on a line perpendicular to the spinal process of L4. After identification of a motor response from the quadriceps, the intensity of the current was reduced to 0.35 mA and 0.5% ropivacaine (39 mL) was administered. During the injection, there were intermittent contractions of the quadriceps. After the block, the patient presented apnea, hypotension, and both pupils were dilated. At the end of the surgery, the patient presented motor block of the lower extremities, which reversed only nine hours after the block. In the postoperative period, the patient complained of severe pain; he was discharged 12 days after the surgery without motor or sensitive deficits. To identify the psoas compartment, where the lumbar plexus blockade is located, the intensity of the current must be between 0.5 and 1 mA. Motor response with low current indicates that the needle may be inside the sheath that surrounds the nervous root and extends to the epidural and subarachnoid spaces, to where the anesthetic solution might spread. Despite the wide safety margin of the procedure, the anesthesiologist must have keen anatomy knowledge, training on the technique, and be constantly alert to perform a lumbar plexus blockade.


A probabilistic model of glenohumeral external rotation strength for healthy normals and rotator cuff tear cases.
The reigning paradigm of musculoskeletal modeling is to construct deterministic models from parameters of an "average" subject and make predictions for muscle forces and joint torques with this model. This approach is limited because it does not perform well for outliers, and it does not model the effects of population parameter variability. The purpose of this study was to simulate variability in musculoskeletal parameters on glenohumeral external rotation strength in healthy normals, and in rotator cuff tear case using a Monte Carlo model. The goal was to determine if variability in musculoskeletal parameters could quantifiably explain variability in glenohumeral external rotation strength. Multivariate Gamma distributions for musculoskeletal architecture and moment arm were constructed from empirical data. Gamma distributions of measured joint strength were constructed. Parameters were sampled from the distributions and input to the model to predict muscle forces and joint torques. The model predicted measured joint torques for healthy normals, subjects with supraspinatus tears, and subjects with infraspinatus-supraspinatus tears with small error. Muscle forces for the three conditions were predicted and compared. Variability in measured torques can be explained by differences in parameter variability.


Trends in tropical tree growth: re-analyses confirm earlier findings.
In a recent Opinion article, Brienen et al. (2016) raise doubts about our finding that tropical tree growth has not increased during 150 years of CO2 rise (Groenendijk et al., 2015; van der Sleen et al., 2015). They claim that our tree-ring data contain evidence for historical growth stimulation that was concealed due to failing regeneration in several species. Here we show that (i) the correction method proposed by Brienen et al. induces a bias towards finding positive growth trends, (ii) the results of Brienen et al. rest on selective removal of species, (iii) there is a simple and effective way to accommodate effects of recruitment failure by subsetting data, and (iv) the application of this method confirms our earlier findings. Thus, our results are robust to effects of recruitment failure and our conclusions remain unchanged: we find no evidence for historical growth changes in our studied tree species.


The ADHD and sleep conundrum: a review.
The relationship between attention-deficit hyperactivity disorder (ADHD) and sleep is a complex one that poses many challenges in clinical practice. Recent studies have helped to elucidate the nature of the brain mechanisms and neuromodulator systems underlying the theoretical associations among sleepiness, arousal, and attention. Studies of sleep disturbances in children with academic and behavioral problems have also underscored the role that primary sleep disorders such as obstructive sleep apnea hypopnea syndrome play in the clinical presentation of symptoms of inattention and behavioral dysregulation. In addition, new methodologies used in examining sleep and sleep patterns in children diagnosed with ADHD have shed further light on the prevalence, type, risk factors for, and impact of sleep disturbances in these children. The following discussion of the multilevel relationships among sleep quality and quantity, neurobehavioral functioning, and the clinical syndrome of ADHD expands on previous reviews of the literature and synthesizes what is currently known about the interaction of sleep and attention/arousal in children to propose possible underlying mechanisms, integrate more recent findings, and highlight important areas for future study. In addition, guidelines are provided for a clinical approach to evaluation and management of children with ADHD and sleep problems.


Implementation of an innovative web-based conference table for community-dwelling frail older people, their informal caregivers and professionals: a process evaluation.
Due to fragmentation of care, continuity of care is often limited in the care provided to frail older people. Further, frail older people are not always enabled to become involved in their own care. Therefore, we developed the Health and Welfare Information Portal (ZWIP), a shared Electronic Health Record combined with a communication tool for community-dwelling frail older people and primary care professionals. This article describes the process evaluation of its implementation, and aims to establish (1) the outcomes of the implementation process, (2) which implementation strategies and barriers and facilitators contributed to these outcomes, and (3) how its future implementation could be improved. Mixed methods study, consisting of (1) a survey among professionals (n = 118) and monitoring the use of the ZWIP by frail older people and professionals, followed by (2) semi-structured interviews with purposively selected professionals (n = 12). 290 frail older people and 169 professionals participated in the ZWIP. At the end of the implementation period, 55% of frail older people and informal caregivers, and 84% of professionals had logged on to their ZWIP at least once. For professionals, the exposure to the implementation strategies was generally as planned, they considered the interprofessional educational program and the helpdesk very important strategies. However, frail older people's exposure to the implementation strategies was less than intended. Facilitators for the ZWIP were the perceived need to enhance interprofessional collaboration and the ZWIP application being user-friendly. Barriers included the low computer-literacy of frail older people, a preference for personal communication and limited use of the ZWIP by other professionals and frail older people. Interviewees recommended using the ZWIP for other target populations as well and adding further strategies that may help frail older people to feel more comfortable with computers and the ZWIP. This study describes the implementation process of an innovative e-health intervention for community-dwelling frail older people, informal caregivers and primary care professionals. As e-health is an important medium for overcoming fragmentation of healthcare and facilitating patient involvement, but its adoption in everyday practice remains a challenge, the positive results of this implementation are promising.


Novel Mutations in FA2H-Associated Neurodegeneration: An Underrecognized Condition?
Hereditary spastic paraplegias and related genetically heterogeneous disorders may be difficult to distinguish clinically. The FA2H gene has been associated with autosomal recessive neurodegenerative phenotypes encompassing spastic paraplegia with or without dystonia, and demyelinating leukodystrophy. To date, few individuals with mutations in the FA2H gene have been described. We report a 5-year-old girl of mixed Filipino and Vietnamese origin who presented with progressive lower limb spasticity and periventricular leukomalacia. The clinical diagnosis of FA2H-associated neurodegeneration was confirmed on the basis of 2 novel mutations in compound heterozygosity in the FA2H gene (p.S70L/p.P323L). This family highlights that FA2H-associated disorders may be underrecognized in children with neurodegeneration of many different ethnicities. Magnetic resonance imaging features play an important role as diagnostic clues in this and other hereditary spastic paraplegias. The consideration of this diagnosis is essential in providing families with important information on prognosis, as well as accurate genetic counseling.


Retinoid signaling determines germ cell fate in mice.
Germ cells in the mouse embryo can develop as oocytes or spermatogonia, depending on molecular cues that have not been identified. We found that retinoic acid, produced by mesonephroi of both sexes, causes germ cells in the ovary to enter meiosis and initiate oogenesis. Meiosis is retarded in the fetal testis by the action of the retinoid-degrading enzyme CYP26B1, ultimately leading to spermatogenesis. In testes of Cyp26b1-knockout mouse embryos, germ cells enter meiosis precociously, as if in a normal ovary. Thus, precise regulation of retinoid levels during fetal gonad development provides the molecular control mechanism that specifies germ cell fate.


Evidence for glomerular actions of epidermal growth factor in the rat.
Epidermal growth factor (EGF), an endogenous mitogenic peptide, has recently been shown to be a potent vasoconstrictor of vascular smooth muscle. In view of its potential role in proliferative and inflammatory renal glomerular diseases, we examined the effects of EGF both on cultured rat mesangial cells and on in vivo glomerular hemodynamics. Mesangial cells possess specific, saturable EGF receptors of differing affinities, with Kd's of 0.1 and 1.7 nM, respectively. EGF produced a rapid increase in intracellular pH of 0.12 +/- 0.01 pH U, which was sodium dependent and amiloride inhibitable. The addition of EGF to mesangial cells cultured on either glass or dimethylpolysiloxane substratum induced reproducible cell contraction. Intrarenal EGF infusion did not affect systemic blood pressure or hematocrit but reversibly decreased GFR and renal blood flow from 4.19 +/- 0.33 to 3.33 +/- 0.26 and from 1.17 +/- 0.09 to 0.69 +/- 0.07 ml/min, respectively. Glomerular micropuncture confirmed decreases in single nephron plasma flow and in single nephron GFR (from 142 +/- 9 to 98 +/- 8 and from 51.6 +/- 11.7 to 28.5 +/- 3.5 nl/min, respectively) which were due to significant increases in both pre- and postglomerular arteriolar resistances (from 1.97 +/- 0.31 to 2.65 +/- 0.36 and from 1.19 +/- 0.11 to 2.00 +/- 0.15 10(10) dyn.s.cm-5 respectively) and to a significant decrease in the ultrafiltration coefficient, Kf, which fell from 0.100 +/- 0.019 to 0.031 +/- 0.007 nl/(s.mmHg). These studies demonstrate that mesangial cells possess specific receptors for EGF, and exposure of these cells to physiologic concentrations of EGF results in an in vitro functional response characterized by activation of Na+/H+ exchange and by resultant intracellular alkalinization, as well as by cell contraction. EGF administration in vivo significantly reduces the glomerular capillary ultrafiltration coefficient, Kf, which, in combination with EGF-induced constriction of both preglomerular and postglomerular arterioles, results in acute major reductions in the rates of glomerular filtration and perfusion.


Land use in relation to composition and abundance of phlebotomines (Diptera: Psychodidae) in five foci of domiciliary transmission of cutaneous leishmaniasis in the Andean region of Colombia.
American cutaneous leishmaniasis is a public health concern in Colombia, its incidence being sustained or focally increased principally by the emergence of domestic transmission concomitantly with the adaptation of the phlebotomine vectors to habitat transformation around households. The objective of the study is to scale up a rapid characterization methodology for evaluating the relationship of land use around the house to the composition and abundance of phlebotomines. Five sites with a history of domestic leishmaniasis transmission in the Andean area of Colombia were selected. The peri-domestic habitat was evaluated at 10 m intervals along eight radial transects, centered on each house, at 45° intervals using a web pattern. Phlebotomines were captured by placing three CDC light traps over two nights both indoors and outdoors (10 m from the house). Blood source and infection were determined by PCR. Spearman rank correlation coefficients and negative binomial regression were used to quantify associations between the phlebotomine abundance and habitat categories. The study demonstrated that the vectors were largely anthropophagic (62% of 79 were human blood) and that a single species in each site was favored by the pertaining agriculture monoculture. Specifically, Pintomyia (Pifanomyia) quasitownsendi was associated with sugar cane in Novillero and La Esmeralda; while Pi. (Pif.) longiflocosa was associated with coffee plantations in Agua Bonita and El Cucal. Honda Alta had a more diverse array of land use and forest coverage with a lower number of specimens but higher species diversity. In terms of distance from the house to an area of a given land use, the abundance of Pi. (Pif.) quasitownsendi was inversely related to the distance to sugar cane plantation (Spearman correlation coefficient, ρ = -0.56, p < 0.001 for outdoor catches, and ρ = -0.50, p < 0.001 indoors). A similar inverse relationship was observed for Pi. (Pif.) longiflocosa with regard to technified coffee (ρ = -0.51, p < 0.001 outdoors, and ρ = -0.48, p < 0.001 indoors). This rapid characterization methodology could guide public heath decision makers in identifying those houses at higher risk of domestic transmission, and also educate farmers to increase the distance between their crops and any neighboring houses.


The shifting biology of prions.
Transmissible spongiform encephalopathies (TSEs), or prion diseases, are rare fatal neurodegenerative diseases of humans and animals. Although some TSEs, like scrapie in sheep, have been known to exist for centuries, bovine spongiform encephalopathy (BSE) was recognized only 15 years ago. New variant Creutzfeldt-Jakob disease (nvCJD) of humans is probably caused by consumption of BSE-infected materials. The nature of the infectious agent is not fully elucidated, but substantial evidence suggests that it is devoid of nucleic acids and consists at least in part of an abnormal form of a host protein termed PrP(C). Despite their rarity, prion diseases have become an important topic in public health and basic research because of the connection between nvCJD and BSE and also because of the unusual biological attributes of the infectious agent.


3-Cyano-3-aza-β-amino Acid Derivatives as Inhibitors of Human Cysteine Cathepsins.
Nitrile-type inhibitors are known to interact with cysteine proteases in a covalent-reversible manner. The chemotype of 3-cyano-3-aza-β-amino acid derivatives was designed in which the N-cyano group is centrally arranged in the molecule to allow for interactions with the nonprimed and primed binding regions of the target enzymes. These compounds were evaluated as inhibitors of the human cysteine cathepsins K, S, B, and L. They exhibited slow-binding behavior and were found to be exceptionally potent, in particular toward cathepsin K, with second-order rate constants up to 52 900 × 10(3) M(-1) s(-1).


Effects of the pneumoperitoneum and Trendelenburg position on respiratory mechanics in the rats by the end-inflation occlusion method.
To describe the consequences of the cranial displacement of the diaphgram occurring during pneumoperitoneum (Pnp) and/or Trendelenburg (Tnd) position on respiratory mechanics. Possible addictive effects and the changes of the viscoelastic respiratory system resistance were studied, which were not extensively described before. The end-inflation occlusion method was applied on eight rats. It allows us to determine mechanical parameters such as respiratory system static elastance, the ohmic resistance due to frictional forces in the airways, and the additional viscoelastic impedance due to tissues deformation. Measurements during mechanical ventilation were taken in controls (supine position), after 20-25° head-down tilting (Tnd), after abdominal air insufflation up to 12 mmHg abdominal pressure in the supine position (Pnp), and combining Tnd + Pnp. Tnd and Pnp modalities were similar to those commonly applied during surgical procedures in humans. We confirmed the previously described detrimental effects on respiratory mechanics due to the diaphgram displacement during both Pnp and Tnd. The increment in the total resistive pressure dissipation was found to depend primarily on the effects on the viscoelastic characteristics of the respiratory system. Data suggesting greater effects of Pnp compared to those of Tnd were obtained. The cranial displacement of the diaphgram occurring as a consequence of Pnp and/or Tnd, for example during laparoscopic surgical procedures, causes an increment of respiratory system elastance and viscoelastic resistance. The analysis of addictive effects show that these are more likely to occur when Pnp + Tnd are compared to isolated Tnd rather than to isolated Pnp.


When two trees go to war.
Rooted phylogenetic networks are used to model non-treelike evolutionary histories. Such networks are often constructed by combining trees, clusters, triplets or characters into a single network that in some well-defined sense simultaneously represents them all. We review these four models and investigate how they are related. Motivated by the parsimony principle, one often aims to construct a network that contains as few reticulations (non-treelike evolutionary events) as possible. In general, the model chosen influences the minimum number of reticulation events required. However, when one obtains the input data from two binary (i.e. fully resolved) trees, we show that the minimum number of reticulations is independent of the model. The number of reticulations necessary to represent the trees, triplets, clusters (in the softwired sense) and characters (with unrestricted multiple crossover recombination) are all equal. Furthermore, we show that these results also hold when not the number of reticulations but the level of the constructed network is minimised. We use these unification results to settle several computational complexity questions that have been open in the field for some time. We also give explicit examples to show that already for data obtained from three binary trees the models begin to diverge.


Basal ganglia-cortical interactions in Parkinsonian patients.
Parkinson's disease is a common and debilitating condition, caused by aberrant activity in a complex basal ganglia-thalamocortical circuit. Therapeutic advances rely on characterising interactions in this circuit. However, recording electrophysiological responses over the entire circuit is impractical. Dynamic causal modelling offers large-scale models of predictive value based on a limited or partial sampling of complex networks. Using dynamic causal modelling, we determined the network changes underlying the pathological excess of beta oscillations that characterise the Parkinsonian state. We modelled data from five patients undergoing surgery for deep brain stimulation of more than one target. We found that connections to and from the subthalamic nucleus were strengthened and promoted beta synchrony, in the untreated compared to the treated Parkinsonian state. Dynamic causal modelling was able to replicate the effects of lesioning this nucleus and may provide a new means of directing the search for therapeutic targets.


Critical Review on Suicide Among Nurses.
Background: Research shows that there is a high prevalence of suicide among nurses. Despite this, it has been 15 years since the last literature review on the subject was published. Aim: The aim of this article is to review the knowledge currently available on the risk of suicide among nurses and on contributory risk factors. Method: A search was conducted in electronic databases using keywords related to prevalence and risk factors of suicide among nurses. The abstracts were analyzed by reviewers according to selection criteria. Selected articles were submitted to a full-text review and their key elements were summarized. Results: Only nine articles were eligible for inclusion in this review. The results of this literature review highlight both the troubling high prevalence of suicide among nurses as well as the persistent lack of studies that examine this issue. Conclusion: Considering that the effects of several factors related to nurses' work and work settings are associated with high stress, distress, or psychiatric problems, we highlight the relevance of investigating work-related factors associated with nurses' risk of suicide. Several avenues for future studies are discussed as well as possible research methods.


Investigation of damage mechanisms of KTiOPO4 crystals by use of a continuous-wave argon laser.
We have developed a simple method that uses cw irradiation from an argon laser to measure the damage threshold of KTP crystals. Our experimental results show that there are two types of damage in KTP crystal, depending on the polarization of the incident laser beam. One type of optical damage is the appearance of gray tracks, including both dark tracks and orange dots, when the pump polarization is perpendicular to the z axis. The other type is invisible damage when the polarization is parallel to the z axis. In addition, we have also observed weak photorefractive two-wave mixing in KTP crystals in each of these polarization states. After a systematic analysis we have concluded that the first type of damage is due to the formation and absorption of Ti3+ and Fe3+ centers and the second is due to the drift of K+ ions. The weak photorefractive effect is the result of screening of the electric field by the drifted K+ ions.


The efficacy of an augmented virtual reality system to alleviate pain in children undergoing burns dressing changes: a randomised controlled trial.
In children, the pain and anxiety associated with acute burn dressing changes can be severe, with drug treatment alone frequently proving to be inadequate. Virtual reality (VR) systems have been successfully trialled in limited numbers of adult and paediatric burn patients. Augmented reality (AR) differs from VR in that it overlays virtual images onto the physical world, instead of creating a complete virtual world. This prospective randomised controlled trial investigated the use of AR as an adjunct to analgesia and sedation in children with acute burns. Forty-two children (30 male and 12 female), with an age range of 3-14 years (median age 9 years) and a total burn surface area ranging from 1 to 16% were randomised into a treatment (AR) arm and a control (basic cognitive therapy) arm after administration of analgesia and/or sedation. Pain scores, pulse rates (PR), respiratory rates (RR) and oxygen saturations (SaO2) were recorded pre-procedurally, at 10 min intervals and post-procedurally. Parents were also asked to grade their child's overall pain score for the dressing change. Mean pain scores were significantly lower (p=0.0060) in the AR group compared to the control group, as were parental pain assessment scores (p=0.015). Respiratory and pulse rates showed significant changes over time within groups, however, these were not significantly different between the two study groups. Oxygen saturation did not differ significantly over time or between the two study groups. This trial shows that augmented reality is a useful adjunct to pharmacological analgesia.


General report & recommendations in predictive, preventive and personalised medicine 2012: white paper of the European Association for Predictive, Preventive and Personalised Medicine.
This report is the collective product of word-leading experts working in the branches of integrative medicine by predictive, preventive and personalised medicine (PPPM) under the coordination of the European Association for Predictive, Preventive and Personalised Medicine. The general report has been prepared as the consortium document proposed at the EPMA World Congress 2011 which took place in Bonn, Germany. This forum analyzed the overall deficits and trends relevant for the top-science and daily practice in PPPM focused on the patient. Follow-up consultations resulted in a package of recommendations for consideration by research units, educators, healthcare industry, policy-makers, and funding bodies to cover the current knowledge deficit in the field and to introduce integrative approaches for advanced diagnostics, targeted prevention, treatments tailored to the person and cost-effective healthcare.


Acceptability of adolescents' occlusion in Finnish municipal health centres with differing timing of orthodontic treatment.
The objective of the study was to compare the acceptability of occlusion among orthodontically treated and untreated adolescents in eight Finnish municipal health centres applying different timing of treatment. A random sample of 16- and 18-year olds (n = 2325) living in these municipalities was invited for a clinical examination, and 1109 adolescents participated. Two calibrated orthodontists blindly examined the participants for the acceptability of occlusion with the Occlusal Morphology and Function Index. The history of orthodontic treatment was elicited by questionnaire. The impact of the history and timing of treatment on the acceptability of occlusion was analysed with logistic regression analysis. The history of orthodontic treatment decreased the odds for acceptability of morphology [odds ratio (OR) = 0.719, 95 per cent confidence limit (CL), P = 0.016] and acceptability of function (OR = 0.724, 95 per cent CL, P = 0.018). The early timing of treatment increased the odds for acceptability of morphology (OR = 1.370, 95 per cent CL, P = 0.042) and of function (OR = 1.420, 95 per cent CL, P = 0.023). No substantial differences were observed in the acceptability of occlusion between the early and late timing health centres. However, the proportion of subjects with acceptable occlusion was slightly higher in the early than in the late timing group. These findings suggest that when examining the effect of timing on treatment outcome, factors other than acceptability of occlusion should be concomitantly evaluated. Consequently, in this context, the duration and cost of treatment need to be investigated.


Increased incidence of peptic ulcer disease in central serous chorioretinopathy patients: a population-based retrospective cohort study.
To investigate peptic ulcer disease and other possible risk factors in patients with central serous chorioretinopathy (CSR) using a population-based database. In this population-based retrospective cohort study, longitudinal data from the Taiwan National Health Insurance Research Database were analyzed. The study cohort comprised 835 patients with CSR and the control cohort comprised 4175 patients without CSR from January 2000 to December 2009. Conditional logistic regression was applied to examine the association of peptic ulcer disease and other possible risk factors for CSR, and stratified Cox regression models were applied to examine whether patients with CSR have an increased chance of peptic ulcer disease and hypertension development. The identifiable risk factors for CSR included peptic ulcer disease (adjusted odd ratio: 1.39, P = 0.001) and higher monthly income (adjusted odd ratio: 1.30, P = 0.006). Patients with CSR also had a significantly higher chance of developing peptic ulcer disease after the diagnosis of CSR (adjusted odd ratio: 1.43, P = 0.009). Peptic ulcer disease and higher monthly income are independent risk factors for CSR. Whereas, patients with CSR also had increased risk for peptic ulcer development.


Serosurveillance study on transmission of H5N1 virus during a 2006 avian influenza epidemic.
In 2006 an outbreak of avian influenza A(H5N1) in Turkey caused 12 human infections, including four deaths. We conducted a serological survey to determine the extent of subclinical infection caused by the outbreak. Single serum samples were collected from five individuals with avian influenza whose nasopharyngeal swabs tested positive for H5 RNA by polymerase chain reaction, 28 family contacts of the cases, 95 poultry cullers, 75 individuals known to have had contact with diseased chickens and 81 individuals living in the region with no known contact with infected chickens and/or patients. Paired serum samples were collected from 97 healthcare workers. All sera were tested for the presence of neutralizing antibodies by enzyme-linked immunoassay, haemagglutination inhibition and microneutralization assays. Only one serum sample, from a parent of an avian influenza patient, tested positive for H5N1 by microneutralization assay. This survey shows that there was minimal subclinical H5N1 infection among contacts of human cases and infected poultry in Turkey in 2006. Further, the low rate of subclinical infection following contact with diseased poultry gave further support to the reported low infectivity of the virus.


The LifeCycle Project-EU Child Cohort Network: a federated analysis infrastructure and harmonized data of more than 250,000 children and parents.
Early life is an important window of opportunity to improve health across the full lifecycle. An accumulating body of evidence suggests that exposure to adverse stressors during early life leads to developmental adaptations, which subsequently affect disease risk in later life. Also, geographical, socio-economic, and ethnic differences are related to health inequalities from early life onwards. To address these important public health challenges, many European pregnancy and childhood cohorts have been established over the last 30 years. The enormous wealth of data of these cohorts has led to important new biological insights and important impact for health from early life onwards. The impact of these cohorts and their data could be further increased by combining data from different cohorts. Combining data will lead to the possibility of identifying smaller effect estimates, and the opportunity to better identify risk groups and risk factors leading to disease across the lifecycle across countries. Also, it enables research on better causal understanding and modelling of life course health trajectories. The EU Child Cohort Network, established by the Horizon2020-funded LifeCycle Project, brings together nineteen pregnancy and childhood cohorts, together including more than 250,000 children and their parents. A large set of variables has been harmonised and standardized across these cohorts. The harmonized data are kept within each institution and can be accessed by external researchers through a shared federated data analysis platform using the R-based platform DataSHIELD, which takes relevant national and international data regulations into account. The EU Child Cohort Network has an open character. All protocols for data harmonization and setting up the data analysis platform are available online. The EU Child Cohort Network creates great opportunities for researchers to use data from different cohorts, during and beyond the LifeCycle Project duration. It also provides a novel model for collaborative research in large research infrastructures with individual-level data. The LifeCycle Project will translate results from research using the EU Child Cohort Network into recommendations for targeted prevention strategies to improve health trajectories for current and future generations by optimizing their earliest phases of life.


Optimal increase in bone mass by continuous local infusion of alendronate during distraction osteogenesis in rabbits.
Several methods have been used to increase bone mass in distraction osteogenesis. Since bone resorption as well as regeneration is stimulated in the distracted segment, bisphosphonate can be a beneficial agent for distraction osteogenesis. Here, we examined the effects of bisphosphonate injected continuously into the regenerate on bone volume, and architectural and mechanical properties of distraction osteogenesis. The left tibia of Japanese White rabbits (n=66) was subjected to slow distraction using an external fixator. At the beginning of the consolidation phase, alendronate (7 microg/kg/day) was infused directly into the lengthened segment for 14 days using an osmotic pump. Control rabbits were infused with phosphate buffered saline (PBS). The tibiae were monitored weekly by soft X-ray and dual-energy X-ray absorptiometry (DXA). The animals were sacrificed at 4, 6, and 8 weeks after operation to examine bone mineral density (BMD) and cortical bone thickness (CBT) by peripheral quantitative computerized tomography (pQCT), while the mechanical property of the lengthened tibia was measured by three-point bending test. In PBS-infused control animals, bone mineral content around the lengthened segment began to decrease after the first week of consolidation phase, forming a tubular bone structure with thin cortex. Infusion of alendronate increased peak bone mineral content around the lengthened segment. At the end of the experiment, volumetric BMD, CBT and mechanical strength of the lengthened segment of the treatment group were approximately twice those of the control animals. Alendronate infused in this manner significantly prevented the osteopenia that critically began early in the consolidation phase, though the dose used in this study was relatively low and no adverse events were noted.


A multi-platform metabolomics approach identifies highly specific biomarkers of bacterial diversity in the vagina of pregnant and non-pregnant women.
Bacterial vaginosis (BV) increases transmission of HIV, enhances the risk of preterm labour, and is associated with malodour. Clinical diagnosis often relies on microscopy, which may not reflect the microbiota composition accurately. We use an untargeted metabolomics approach, whereby we normalize the weight of samples prior to analysis, to obtained precise measurements of metabolites in vaginal fluid. We identify biomarkers for BV with high sensitivity and specificity (AUC = 0.99) in a cohort of 131 pregnant and non-pregnant Rwandan women, and demonstrate that the vaginal metabolome is strongly associated with bacterial diversity. Metabolites associated with high diversity and clinical BV include 2-hydroxyisovalerate and γ-hydroxybutyrate (GHB), but not succinate, which is produced by both Lactobacillus crispatus and BV-associated anaerobes in vitro. Biomarkers associated with high diversity and clinical BV are independent of pregnancy status, and were validated in a blinded replication cohort from Tanzania (n = 45), where we predicted clinical BV with 91% accuracy. Correlations between the metabolome and microbiota identified Gardnerella vaginalis as a putative producer of GHB, and we demonstrate production by this species in vitro. This work illustrates how changes in community structure alter the chemical composition of the vagina, and identifies highly specific biomarkers for a common condition.


Apathy in nursing home residents with dementia: results from a cluster-randomized controlled trial.
Here we evaluate an interdisciplinary occupational and sport therapy intervention for dementia patients suffering from apathy. A prospective, controlled, rater-blinded, clinical trial with two follow-ups was conducted as part of a larger cluster-randomized trial in 18 nursing homes in Berlin. n=117 dementia patients with apathy, defined as a score of 40 or more on the apathy evaluation scale (AES) or presence of apathy on the Neuropsychiatric Inventory (NPI), were randomly assigned to intervention or control group. The intervention included 10 months of brief activities, provided once a week. The primary outcome measure was the total score on the AES scale measured directly after the intervention period and again after 12 months. We found significant group differences with respect to apathy during the 10 month intervention period (F2,82=7.79, P<0.01), which reflected an increase in apathy in the control group, but not in the intervention group. Within one year after the intervention was ceased, the treatment group worsened and no longer differed significantly from the control group (P=0.55). Our intervention was effective for the therapy of apathy in dementia, when applied, but not one year after cessation of therapy.


Mechanical cell-matrix feedback explains pairwise and collective endothelial cell behavior in vitro.
In vitro cultures of endothelial cells are a widely used model system of the collective behavior of endothelial cells during vasculogenesis and angiogenesis. When seeded in an extracellular matrix, endothelial cells can form blood vessel-like structures, including vascular networks and sprouts. Endothelial morphogenesis depends on a large number of chemical and mechanical factors, including the compliancy of the extracellular matrix, the available growth factors, the adhesion of cells to the extracellular matrix, cell-cell signaling, etc. Although various computational models have been proposed to explain the role of each of these biochemical and biomechanical effects, the understanding of the mechanisms underlying in vitro angiogenesis is still incomplete. Most explanations focus on predicting the whole vascular network or sprout from the underlying cell behavior, and do not check if the same model also correctly captures the intermediate scale: the pairwise cell-cell interactions or single cell responses to ECM mechanics. Here we show, using a hybrid cellular Potts and finite element computational model, that a single set of biologically plausible rules describing (a) the contractile forces that endothelial cells exert on the ECM, (b) the resulting strains in the extracellular matrix, and (c) the cellular response to the strains, suffices for reproducing the behavior of individual endothelial cells and the interactions of endothelial cell pairs in compliant matrices. With the same set of rules, the model also reproduces network formation from scattered cells, and sprouting from endothelial spheroids. Combining the present mechanical model with aspects of previously proposed mechanical and chemical models may lead to a more complete understanding of in vitro angiogenesis.


Disk Carbapenemase Test for the Rapid Detection of KPC-, NDM-, and Other Metallo-β-Lactamase-Producing Gram-Negative Bacilli.
Rapid detection of carbapenemase-producing gram-negative bacilli (GNB) is required for optimal treatment of infected patients. We developed and assessed a new disk carbapenemase test (DCT). Paper disks containing 0.3 mg of imipenem and bromothymol blue indicator were developed, and the performance of the DCT were evaluated by using 742 strains of GNB with or without carbapenemases. The paper disks were simple to prepare, and the dried disks were stable at -20°C and at 4°C. The DCT detected 212 of 215 strains (98.6% sensitivity with 95% confidence interval [CI] 96.0-99.5%) of GNB with known class A (KPC and Sme) and class B (NDM, IMP, VIM, and SIM) carbapenemases within 60 min, but failed to detect GES-5 carbapenemase. The DCT also detected all two Escherichia coli isolates with OXA-48, but failed to detect GNB with OXA-232, and other OXA carbapenemases. The DCT showed 100% specificity (95% CI, 99.2-100%) in the test of 448 imipenem-nonsusceptible, but carbapenemase genes not tested, clinical isolates of GNB. The DCT is simple and can be easily performed, even in small laboratories, for the rapid detection of GNB with KPC, NDM and the majority of IMP, VIM, and SIM carbapenemases.


Stimulated Brillouin scattering based in-band OSNR monitoring technique for 40 Gbps and 100 Gbps optical transparent networks.
We demonstrate experimentally and numerically that the SBS based in-band OSNR monitoring technique can be used for dual polarization signals. We also present novel approach for a drastic enhancement of the sensitivity monitoring range by intentionally adding in-band ASE noise into the signal. Numerical results are provided for 44.6 Gbps DPSK, 44.6 Gbps DQPSK and 112 Gbps Dual Polarization (DP-) QPSK signals, with both 100 GHz and 50 GHz channel spacing scenarios.


Solid fuel use, socioeconomic indicators and risk of cardiovascular diseases and all-cause mortality: a prospective cohort study in a rural area of Sichuan, China.
Estimates indicate that household air pollution caused by solid fuel burning accounted for about 1.03 million premature mortalities in China in 2016. In the country's rural areas, more than half the population still relies on biomass fuels and coals for cooking and heating. Understanding the health impact of indoor air pollution and socioeconomic indicators is essential for the country to improve its developmental targets. We aimed to describe demographic and socioeconomic characteristics associated with solid fuel users in a rural area in China. We also estimated the risk of cardiovascular disease and all-cause mortality in association with solid fuel use and described the relationship between solid fuel use, socioeconomic status and mortality. We also measured the risk of long-term use, and the effect of ameliorative action, on mortality caused by cardiovascular disease and other causes. We used the China Kadoorie Biobank (CKB) site in Pengzhou, Sichuan, China. We followed a cohort of 55 687 people over 2004-13. We calculated the mean and standard deviation among subgroups classified by fuel use types: gas, coal, wood and electricity (central heating additionally for heating). We tested the mediation effect using the stepwise method and Sobel test. We used Cox proportional models to estimate the risk of incidences of cardiovascular disease and mortality with survival days as the time scale, adjusted for age, gender, socioeconomic status, physical measurements, lifestyle, stove ventilation and fuel type used for other purposes. The survival days were defined as the follow-up days from the baseline survey till the date of death or 31 December 2013 if right-censored. We also calculated the absolute mortality rate difference (ARD) between the exposure group and the reference group. The study population had an average age of 51.0, and 61.9% of the individuals were female; 64.8% participants (n = 35 543) cooked regularly and 25.4% participants (n = 13 921) needed winter heating. With clean fuel users as the reference group, participant households that used solid fuel for cooking or heating both had a higher risk of all-cause mortality: hazard ratio (HR) for: cooking, 1.11 [95% confidence interval (CI) 1.02, 1.26]; heating, 1.34 (95% CI 1.16, 1.54). Solid fuel used for winter heating was associated with a higher risk of mortality caused by cerebrovascular disease: HR 1.64 (95% CI 1.12, 2.40); stroke: HR 1.70 (95% CI 1.13, 2.56); and cardiovascular disease: HR 1.49 (95% CI 1.10, 2.02). Low income and poor education level had a significant correlation with solid fuel used for cooking: odds ratio (OR) for income: 2.27 (95% CI 2.14, 2.41); education: 2.34 (95% CI 2.18, 2.53); and for heating: income: 2.69 (95% CI 2.46, 2.97); education: 2.05 (95% CI 1.88, 2.26), which may be potential mediators bridging the effects of socioeconomic status factors on cardiovascular disease and all-cause mortality. Solid fuel used for cooking and heating accounted for 42.4% and 81.1% of the effect of poor education and 55.2% and 76.0% of the effect of low income on all-cause mortality, respectively. The risk of all-cause mortality could be ameliorated by stopping regularly cooking and heating using solid fuel or switching from solid fuel to clean fuels: HR for cooking: 0.90 (95% CI 0.84, 0.96); heating: 0.76 (95% CI 0.64, 0.92). Our study reinforces the evidence of an association between solid fuel use and risk of cardiovascular disease and all-cause mortality. We also assessed the effect of socioeconomic status as the potential mediator on mortality. As solid fuel use was a major contributor in the effect of socioeconomic status on cardiovascular disease and all-cause mortality, policies to improve access to clean fuels could reduce morbidity and mortality related to poor education and low income.


Extended Endoscopic Endonasal Approach for Craniopharyngioma Removal.
Objective Endoscopic transsphenoidal extended endoscopic approach (EEA) represents a valid alternative to microsurgery for craniopharyngiomas removal, especially for retrochiasmatic lesions without large parasellar extension. The present video illustrates the salient surgical steps of the EEA for craniopahryngioma removal. Patient A 52-year-old man presented with a bitemporal hemianopia and a bilateral decreased visual acuity. MRI showed a Kassam type III cystic craniopharyngioma with a solid component ( Fig. 1 , panels A and B). Surgical Procedure The head is rotated 10 degrees toward the surgeons. The nasal step is started through the left nostril with a middle turbinectomy. A nasoseptal flap is harvested and positioned in the left choana. The binostril approach allows a large sphenoidotomy to expose the key anatomic landmarks. The craniotomy boundaries are the planum sphenoidale superiorly, the median opticocarotid recesses, the internal carotid artery laterally and the clival recess inferiorly. After dural opening and superior intercavernous sinus coagulation, the tumor is entirely removed ( Fig. 2 , panels A and B). Skull base reconstruction is ensured by fascia lata grafting and nasoseptal flap positioning. Results Postoperative MRI showed the complete tumor resection ( Fig. 1 , panels C and D). At 3 months postoperatively, the bitemporal hemianopia regressed and the visual acuity improved. A novel left homonymous hemianopia developed secondary to optic tract manipulation. Conclusions The extended EEA is a valid surgical approach for craniopharyngioma resection. A comprehensive knowledge of the sellar and parasellar anatomy is mandatory for safe tumor removal with decreased morbidity and satisfactory oncologic results. The link to the video can be found at: https://youtu.be/NrCPPnVK2qA .


Influence of Different Antidepressants on Ocular Surface in Patients With Major Depressive Disorder.
Several studies have previously reported the association between dry eye and depression along with the treatment of depression. The aim of this study was to investigate the effects of different antidepressant drugs on tear parameters in patients with major depressive disorder. We recruited 132 patients who were using different antidepressants and 58 healthy controls. Venlafaxine, duloxetine, escitalopram, and sertraline were used by 34, 28, 36, and 34 patients, respectively. The participants filled out and completed the Beck Depression Scale. We recorded Schirmer test, tear breakup time (TBUT) and corneal staining values of the participants. The Ocular Surface Disease Index was completed by the participants. In addition, we evaluated the tear meniscus parameters by using anterior segment optical coherence tomography. All conventional dry eye tests and tear meniscus parameters were significantly lesser in the depression group than in the control group (Schirmer test, 11.41 ± 6.73 mm and 22.53 ± 4.98 mm; TBUT, 5.29 ± 2.92 seconds and 13.38 ± 1.72; Corneal staining, tear meniscus area, 0.026 ± 0.012 mm2 and 0.11 ± 0.025 mm2; tear meniscus depth, 182.75 ± 78.79 μm and 257.48 ± 90.1 μm; tear meniscus height, 290.3 ± 133.63 μm and 459.78 ± 180.26 μm, in patients and controls, respectively). The tear parameters of the duloxetine group were lowest among the drug groups and Schirmer test, and TBUT of the venlafaxine group was statistically significantly different from the duloxetine group (P = 0.028 and P = 0.017, respectively). Ocular Surface Disease Index score of the depression group was significantly higher than the control group (31.12 ± 21.15 and 17.43 ± 11.75 in depression and control group, respectively.). We found that the usage of selective serotonin reuptake inhibitors and serotonin noradrenaline reuptake inhibitors affects the ocular surface by a mechanism other than the anticholinergic system. Besides serotonin blockage, the noradrenaline blockade of serotonin noradrenaline reuptake inhibitors may increase the dry eye findings on the ocular surface.


Perceptions of family caregivers of cancer patients about the challenges of caregiving: a qualitative study.
The experience of caring for a family member with cancer is associated with several care-related problems and challenges for the caregiver. The comprehensive and in-depth understanding of the trials and tribulations of caregiving can be a step towards resolving the problems faced by family caregivers of these patients. The present study aimed to explore challenges faced by Iranian family caregivers of cancer patients. The present qualitative study was conducted through in-depth semi-structured interviews held with 21 family caregivers of cancer patients selected through purposive sampling. Interviews continued until saturation of data. All interviews were recorded, transcribed and analysed through conventional content analysis. The codes extracted from interviews produced four main themes, including 'confusion', 'uncertainty', 'disintegration' and 'setback', which collectively caused suffering for family caregivers. Care provided in an atmosphere of suffering and discontent diminishes caregiver's quality of life and quality of patient care. Health planners should therefore consider the challenges and sufferings faced by family caregivers and should seek to obviate them through appropriate plans.


Psychological therapies for pathological and problem gambling.
Various psychological therapies for pathological and problem gambling have been evaluated in randomised trials. A synthesis of best-quality evidence is required. The objective was to synthesise evidence from randomised trials of psychological therapies for pathological and problem gambling (cognitive-behaviour therapy (CBT), motivational interviewing therapy, integrative therapy, other psychological therapy), in order to indicate the efficacy of therapies and durability of therapy effects, relative to control conditions. We conducted a search of the Cochrane Depression, Anxiety and Neurosis Review Group's Specialised Register (CCDANCTR), which includes relevant randomised controlled trials from the following bibliographic databases: CENTRAL (The Cochrane Central Register of Controlled Trials) (all years), EMBASE (1974 -), MEDLINE (1950 -) and PsycINFO (1967 -). We also carried out complementary searches of MEDLINE, EMBASE, PsycINFO, LILACS and CENTRAL for studies published between January 1980 and October 2011. We examined the WHO International Clinical Trials Registry Platform and ClinicalTrials.gov and also conducted manual searches of selected journals and reference lists of included studies. Included studies were clinical trials using random allocation to groups, considering pathological or problem gamblers, and evaluating a psychological therapy for pathological or problem gambling. Control conditions included 'no treatment' controls, referral to Gamblers Anonymous and non-specific treatment component controls. We systematically extracted data on the characteristics and results of studies. Primary outcomes were measures of gambling symptom severity, financial loss from gambling and frequency of gambling. Secondary outcomes were occurrence of pathological gambling diagnoses and depression and anxiety symptoms. Treatment effects were defined by comparisons between therapy and control conditions at post-treatment assessments (conducted from 0 to 3 months following completion of treatment) and follow-up assessments (conducted from 9 to 12 months following completion of treatment), respectively, using the standardised mean difference (SMD) or risk ratio (RR). We synthesised results through random-effects meta-analysis. Fourteen studies (n = 1245) met the inclusion criteria. Eleven studies compared CBT with control and comparisons at 0 to 3 months post-treatment showed beneficial effects of therapy that ranged from medium (when defined by financial loss from gambling: SMD -0.52; 95% confidence interval (CI) -0.71 to -0.33, n = 505) to very large (for gambling symptom severity: SMD -1.82; 95% CI -2.61 to -1.02, n = 402). Only one study (n = 147) compared groups at 9 to 12 months follow-up and produced smaller effects that were not significant. Four studies of motivational interviewing therapy were identified and mainly considered samples demonstrating less severe gambling (relative to studies of pathological gamblers). Data suggested reduced financial loss from gambling following motivational interviewing therapy at 0 to 3 months post-treatment (SMD -0.41; 95% CI -0.75 to -0.07, n = 244), although comparisons on other outcomes were not significant. The effect approached zero when defined by gambling symptom severity (SMD -0.03; 95% CI -0.55 to 0.50, n = 163). Studies compared groups at 9 to 12 months follow-up and found a significant effect of motivational interviewing therapy in terms of frequency of gambling (SMD -0.53; 95% CI -1.04 to -0.02, n = 62), with comparisons on other outcomes that were not significant. Two studies of integrative therapies also considered samples demonstrating overall low gambling severity, and found no significant effects of therapy at 0 to 3 months post-treatment. Comparisons at 9 to 12 months follow-up suggested a medium effect from therapy in terms of gambling symptom severity, with no significant differences for other outcomes. One study (n = 18) considered another psychological therapy (i.e.Twelve-Step Facilitated Group Therapy) and suggested beneficial effects in terms of most outcomes at 0 to 3 months post-treatment. The evidence supporting these various classes of therapy ranged from very low to low quality. This review supports the efficacy of CBT in reducing gambling behaviour and other symptoms of pathological and problem gambling immediately following therapy. However, the durability of therapeutic gain is unknown. There is preliminary evidence for some benefits from motivational interviewing therapy in terms of reduced gambling behaviour, although not necessarily other symptoms of pathological and problem gambling. However, the findings are based on few studies and additional research is needed to inform conclusions. There is also evidence suggestive of some possible benefit from integrative therapies, and other psychological therapies for pathological and problem gambling. However, there are too few studies and evidence is insufficient to evaluate these therapies. The majority of studies in this review varied in risk of bias, and much of the evidence comes from studies with multiple limitations. The current data may thus reflect overestimates of treatment efficacy.


Coexpression of connexins 40 and 43 enhances the pH sensitivity of gap junctions: a model for synergistic interactions among connexins.
Gap junctions are formed by oligomerization of a protein called connexin. Most cells express more than one connexin isotype. Atrial myocytes, for example, coexpress connexin (Cx) 40 and Cx43. The consequence of connexin coexpression on the regulation of gap junctions is not well understood. In the present study, we show that cells coexpressing Cx40 and Cx43 are more susceptible to acidification-induced uncoupling than those cells expressing only one connexin isotype. Xenopus oocytes were injected with mRNA for Cx40, Cx43, or a combination of both. Intracellular pH and junctional conductance were simultaneously measured while cells were progressively acidified by superfusion with a bicarbonate-buffered solution gassed with increasing concentrations of carbon dioxide. The data show that the pKa (ie, the pH at which junctional conductance decreased to 50% from maximum) shifted from approximately 6.7 when cells expressed only Cx40 or only Cx43 to approximately 7.0 when one of the oocytes was coexpressing both connexins. Truncation of the carboxyl terminal domains of the connexins caused the loss of pH sensitivity even after coexpression. The data are interpreted on the basis of previous studies from our laboratory that demonstrated heterodomain interactions in the regulation of Cx40 and Cx43 gap junctions. The possible implications of these findings on the regulation of native gap junctions that express both connexins remain to be determined.


Investigating the structure and dynamics of the PIK3CA wild-type and H1047R oncogenic mutant.
The PIK3CA gene is one of the most frequently mutated oncogenes in human cancers. It encodes p110α, the catalytic subunit of phosphatidylinositol 3-kinase alpha (PI3Kα), which activates signaling cascades leading to cell proliferation, survival, and cell growth. The most frequent mutation in PIK3CA is H1047R, which results in enzymatic overactivation. Understanding how the H1047R mutation causes the enhanced activity of the protein in atomic detail is central to developing mutant-specific therapeutics for cancer. To this end, Surface Plasmon Resonance (SPR) experiments and Molecular Dynamics (MD) simulations were carried out for both wild-type (WT) and H1047R mutant proteins. An expanded positive charge distribution on the membrane binding regions of the mutant with respect to the WT protein is observed through MD simulations, which justifies the increased ability of the mutated protein variant to bind to membranes rich in anionic lipids in our SPR experiments. Our results further support an auto-inhibitory role of the C-terminal tail in the WT protein, which is abolished in the mutant protein due to loss of crucial intermolecular interactions. Moreover, Functional Mode Analysis reveals that the H1047R mutation alters the twisting motion of the N-lobe of the kinase domain with respect to the C-lobe and shifts the position of the conserved P-loop residues in the vicinity of the active site. These findings demonstrate the dynamical and structural differences of the two proteins in atomic detail and propose a mechanism of overactivation for the mutant protein. The results may be further utilized for the design of mutant-specific PI3Kα inhibitors that exploit the altered mutant conformation.


CD10+ pancreatic stellate cells enhance the progression of pancreatic cancer.
Pancreatic stellate cells (PSCs) promote the progression of pancreatic cancer by producing extracellular matrix and soluble factors. However, the functional heterogeneity of PSCs has not been identified until now. Detailed characterization of the PSCs in human pancreatic cancer would provide a set of potential targets for stroma-directed therapy. We isolated PSCs from fresh pancreatic ductal adenocarcinoma tissue and sorted them by flow cytometry according to cell surface expression of CD10, which is a stromal prognostic marker for various tumors. We analyzed the functional differences between CD10(+) PSCs and CD10(-) PSCs. Immunohistochemical analysis showed that the frequency of CD10 expression by PSCs was markedly higher in tumor tissue than in normal tissue (33.7% vs 0%, respectively, P = .028). In pancreatic ductal adenocarcinoma, CD10 expression by PSCs was associated with positive nodal metastases (P = .011) and a shorter survival time (P < .001). In vitro coculture experiments showed that CD10(+) PSCs promoted the invasiveness of pancreatic cancer cell lines, SUIT-2 and Panc-1 cells more intensively than CD10(-) PSCs. CD10(+) PSCs significantly increased the tumor growth and invasiveness of SUIT-2 cells in a murine cotransplantation model. CD10(+) PSCs secreted higher levels of matrix metalloproteinase 3 than CD10(-) PSCs, and knockdown of matrix metalloproteinase 3 in cocultured PSCs reduced the invasion of SUIT-2 and Panc-1 cells. CD10(+) PSCs enhance the progression of pancreatic cancer cells. CD10(+) PSCs may be a candidate for selective therapeutic targeting in the treatment of pancreatic cancer.


The Warfarin and Antiplatelet Therapy in Heart Failure trial (WATCH): rationale, design, and baseline patient characteristics.
The role of anticoagulation in patients with chronic heart failure has long been an area of interest and controversy. Traditionally the goal of anticoagulation has been to prevent embolic events, but recent trials also demonstrated that oral anticoagulation also prevents vascular events in patients with prior myocardial infarction, who constitute the majority of heart failure patients. Although antiplatelet agents also reduce postinfarction vascular events, few data are available in heart failure patients, and some evidence suggests that aspirin may also have the potential to worsen heart failure morbidity and mortality, possibly by interfering with the effects of angiotensin-converting enzyme inhibitors. Methods and results The Warfarin and Antiplatelet Therapy in Chronic Heart Failure (WATCH) trial was undertaken to determine the optimal antithrombotic agent for heart failure patients. WATCH was a prospective-randomized trial in which symptomatic heart failure patients in sinus rhythm with ejection fractions < or =35% taking angiotensin-converting enzyme inhibitors (unless not tolerated) and diuretics were randomized to open-label warfarin (target International Normalized Ratio 2.5-3.0) or double-blind antiplatelet therapy with aspirin 162 mg or clopidogrel 75 mg. Two primary comparisons were specified: anticoagulation with warfarin versus antiplatelet therapy with aspirin and antiplatelet therapy with clopidogrel versus antiplatelet therapy with aspirin. The primary outcome is the composite of death from all causes, nonfatal myocardial infarction, and nonfatal stroke analyzed as time to first event using the intent-to-treat approach. The secondary endpoint was the broader composite of death from all causes, nonfatal myocardial infarction, non-fatal stroke, and hospitalizations for worsening heart failure, unstable angina pectoris, and systemic or pulmonary artery embolic events. Additional prespecified analyses include heart failure events, coronary events, and resource utilization. Although the trial was designed to enter 4500 patients, it was terminated 18 months prematurely in June 2003 by the VA Cooperative Study Program because of poor enrollment with a resulting reduction of its power to achieve its original objective. This manuscript describes the study rationale, protocol design, and the baseline characteristics of the 1587 patients who were entered into the study. The WATCH trial will help define the optimal approach to antithrombotic therapy in the contemporary management of patients with chronic heart failure resulting from left ventricular systolic dysfunction.


Sexual health in primary health care - a qualitative study of nurses' experiences.
To illuminate nurses' experiences and opportunities to discuss sexual health with patients in primary health care. Sexual health is a concept associated with many taboos, and research shows that nurses feel uncomfortable talking to patients about sexual health and therefore avoid it. This avoidance forms a barrier between patient and nurse which prevents nurses from giving satisfactory health care to patients. A qualitative descriptive design. Semi-structured interviews were conducted with nine nurses in primary health care in Sweden. Data were analysed using qualitative content analysis. During the analysis phase, five subcategories and two main categories were identified. The two main categories were as follows: 'factors that influence nurses' opportunities of talking to patients about sexual health' and 'nurses' experiences of talking to patients about sexual health'. Social norms in society were an obstacle for health professionals' opportunities to feel comfortable and act professionally. The nurses' personal attitude and knowledge were of great significance in determining whether they brought up the topic of sexual health or not. The nurses found it easier to bring up the topic of sexual health with middle-aged men with, for example, diabetes. One reason for this is that they found it easier to talk to male patients. A further reason is the fact that they had received training in discussing matters of sexual health in relation to diabetes and other conditions affecting sexual health. Nurses in primary care express the necessity of additional education and knowledge on the subject of sexual health. The healthcare organisation must be reformed to put focus on sexual health. Guidelines for addressing the topic of sexual health must be implemented to establish conditions that will increase nurse's knowledge and provide them with the necessary tools for discussing sexual health with patients.


Neuronal surface and glutamic acid decarboxylase autoantibodies in Nonparaneoplastic stiff person syndrome.
High titers of autoantibodies to glutamic acid decarboxylase (GAD) are well documented in association with stiff person syndrome (SPS). Glutamic acid decarboxylase is the rate-limiting enzyme in the synthesis of γ-aminobutyric acid (GABA), and impaired function of GABAergic neurons has been implicated in the pathogenesis of SPS. Autoantibodies to GAD might be the causative agent or a disease marker. To investigate the characteristics and potential pathogenicity of GAD autoantibodies in patients with SPS and related disorders. Retrospective cohort study and laboratory investigation. Weatherall Institute of Molecular Medicine, University of Oxford. Twenty-five patients with SPS and related conditions identified from the Neuroimmunology Service. Neurological examination, serological characterization and experimental studies. Characterization of serum GAD antibodies from patients with SPS and evidence for potential pathogenicity. We detected GAD autoantibodies at a very high titer (median, 7500 U/mL) in 19 patients (76%), including all 12 patients with classic SPS. The GAD autoantibodies were high affinity (antibody dissociation constant, 0.06-0.78 nmol) and predominantly IgG1 subclass. The patients' autoantibodies co-localized with GAD on immunohistochemistry and in permeabilized cultured cerebellar GABAergic neurons, as expected, but they also bound to the cell surface of unpermeabilized GABAergic neurons. Adsorption of the highest titer (700 000 U/mL) serum with recombinant GAD indicated that these neuronal surface antibodies were not directed against GAD itself. Although intraperitoneal injection of IgG purified from the 2 available GAD autoantibody–ositive purified IgG preparations did not produce clinical or pathological evidence of disease, SPS and control IgG were detected in specific regions of the mouse central nervous system, particularly around the lateral and fourth ventricles. Autoantibodies to GAD are associated with antibodies that bind to the surface of GABAergic neurons and that could be pathogenic. Moreover, in mice, human IgG from the periphery gained access to relevant areas in the hippocampus and brainstem. Identification of the target of the non-GAD antibodies and peripheral and intrathecal transfer protocols, combined with adsorption studies, should be used to demonstrate the role of the non-GAD IgG in SPS.


Detecting order and lateral pressure at biomimetic interfaces using a mechanosensitive second-harmonic-generation probe.
A planarizable push-pull molecular probe with mechanosensitive properties was investigated at several biomimetic interfaces, consisting of different phospholipid monolayers located between dodecane and an aqueous buffer solution, using the interface-specific surface-second-harmonic-generation (SSHG) technique. Whereas the SSHG spectra recorded at liquid-disordered interfaces were similar to the absorption spectra in bulk solutions, those measured at liquid-ordered phases exhibited a remarkable shift towards lower energies to an extent depending on the surface pressure of the phospholipid monolayer. On the basis of quantum-chemical calculations, this effect was accounted for by the planarization of the mechanosensitive probe. Polarization-resolved SSHG measurements revealed that the average orientation of the probe at the interface is an even more sensitive reporter of lateral pressure and order than the spectral shape. Additionally, time-resolved SSHG measurements pointed to slower dynamics upon intercalation inside the phospholipid monolayer, most likely due to the more constrained environment. This study demonstrates that the concept of mechanosensitive optical probes can be further exploited when combined with a surface-selective nonlinear optical technique.


Internal gallbladder drainage prevents development of acute cholecystitis in a pig model: a randomized study.
Acute cholecystitis can be the result of retention of bile in the gallbladder with possible secondary infection and ischaemia. The aim of the present study was to investigate whether internal drainage of the gallbladder could protect against the development of acute cholecystitis in a pig model. Twenty pigs were randomized to either internal drainage (drained) or not (undrained). Day 0 acute cholecystitis was induced by ligation of the cystic artery and duct together with inoculation of bacteria. Four days later the pigs were killed and the gallbladders were removed and histologically scored for the presence of cholecystitis. Bile and blood samples were collected for bacterial culturing and biochemical analyses. The histological examination demonstrated statistical significant differences in acute cholecystitis development between groups, the degree of inflammation being highest in undrained pigs. There were no differences in bacterial cultures between the two groups. Internal drainage of the gallbladder protected against the development of acute cholecystitis in the present pig model. These findings support the theory that gallstone impaction of the cystic duct plays a crucial role as a pathogenetic mechanism in the development of acute cholecystitis and suggest that internal drainage may be a way to prevent and treat acute cholecystitis.


The Effect of Binaural Beats on Visuospatial Working Memory and Cortical Connectivity.
Binaural beats utilize a phenomenon that occurs within the cortex when two different frequencies are presented separately to each ear. This procedure produces a third phantom binaural beat, whose frequency is equal to the difference of the two presented tones and which can be manipulated for non-invasive brain stimulation. The effects of binaural beats on working memory, the system in control of temporary retention and online organization of thoughts for successful goal directed behavior, have not been well studied. Furthermore, no studies have evaluated the effects of binaural beats on brain connectivity during working memory tasks. In this study, we determined the effects of different acoustic stimulation conditions on participant response accuracy and cortical network topology, as measured by EEG recordings, during a visuospatial working memory task. Three acoustic stimulation control conditions and three binaural beat stimulation conditions were used: None, Pure Tone, Classical Music, 5Hz binaural beats, 10Hz binaural beats, and 15Hz binaural beats. We found that listening to 15Hz binaural beats during a visuospatial working memory task not only increased the response accuracy, but also modified the strengths of the cortical networks during the task. The three auditory control conditions and the 5Hz and 10Hz binaural beats all decreased accuracy. Based on graphical network analyses, the cortical activity during 15Hz binaural beats produced networks characteristic of high information transfer with consistent connection strengths throughout the visuospatial working memory task.