Kinetic investigation of human 5-lipoxygenase with arachidonic acid.
Human 5-lipoxygenase (5-LOX) is responsible for the formation of leukotriene (LT)A4, a pivotal intermediate in the biosynthesis of the leukotrienes, a family of proinflammatory lipid mediators. 5-LOX has thus gained attention as a potential drug target. However, details of the kinetic mechanism of 5-LOX are still obscure. In this Letter, we investigated the kinetic isotope effect (KIE) of 5-LOX with its physiological substrate, arachidonic acid (AA). The observed KIE is 20±4 on kcat and 17±2 on kcat/KM at 25°C indicating a non-classical reaction mechanism. The observed rates show slight temperature dependence at ambient temperatures ranging from 4 to 35°C. Also, we observed low Arrhenius prefactor ratio (AH/AD=0.21) and a small change in activation energy (Ea(D)-Ea(H)=3.6J/mol) which suggests that 5-LOX catalysis involves tunneling as a mechanism of H-transfer. The measured KIE for 5-LOX involves a change in regioselectivity in response to deuteration at position C7, resulting in H-abstraction form C10 and formation of 8-HETE. The viscosity experiments influence the (H)kcat, but not (D)kcat. However the overall kcat/KM is not affected for labeled or unlabeled AA, suggesting that either the product release or conformational rearrangement might be involved in dictating kinetics of 5-LOX at saturating conditions. Investigation of available crystal structures suggests the role of active site residues (F421, Q363 and L368) in regulating the donor-acceptor distances, thus affecting H-transfer as well as regiospecificity. In summary, our study shows that that the H-abstraction is the rate limiting step for 5-LOX and that the observed KIE of 5-LOX is masked by a change in regioselectivity.


A colorimetric assay to rapidly determine the activities of lytic polysaccharide monooxygenases.
Lytic polysaccharide monooxygenase (LPMOs) are enzymes that catalyze the breakdown of polysaccharides in biomass and have excellent potential for biorefinery applications. However, their activities are relatively low, and methods to measure these activities are costly, tedious or often reflect only an apparent activity to the polysaccharide substrates. Here, we describe a new method we have developed that is simple to use to determine the activities of type-1 (C1-oxidizing) LPMOs. The method is based on quantifying the ionic binding of cations to carboxyl groups formed by the action of type-1 LPMOs on polysaccharides. It allows comparisons to be made of activities under different conditions. Based on the colorimetric detection and quantification of the pyrocatechol violet (PV)-Ni2+ complex, we have developed an assay to reliably detect and quantify carboxylate moieties introduced by type-1 LPMOs. Conditions were optimized for determining the activities of specific LPMOs. Comparisons were made of the activities against cellulose and chitin of a novel AA10 LPMO and a recently reported family AA11 LPMO. Activities of both LPMOs were boosted by hydrogen peroxide in the 1st hour of the reaction, with a 16-fold increase for the family AA11 LPMO, and up to a 34-fold increase for the family AA10 LPMO. We developed a versatile colorimetric cation-based assay to determine the activities of type-1 LPMOs. The assay is quick, low cost and could be adapted for use in industrial biorefineries.


Hybrid nanostructures for SERS: materials development and chemical detection.
This review focuses on recent developments in hybrid and nanostructured substrates for SERS (surface-enhanced Raman scattering) studies. Thus substrates composed of at least two distinct types of materials, in which one is a SERS active metal, are considered here aiming at their use as platforms for chemical detection in a variety of contexts. Fundamental aspects related to the SERS effect and plasmonic behaviour of nanometals are briefly introduced. The materials described include polymer nanocomposites containing metal nanoparticles and coupled inorganic nanophases. Chemical approaches to tailor the morphological features of these substrates in order to get high SERS activity are reviewed. Finally, some perspectives for practical applications in the context of chemical detection of analytes using such hybrid platforms are presented.


Relationship Between Tongue Base Region Pressures and Vallecular Clearance.
Tongue base pressures have been thought to provide primary bolus clearance through the pharynx during swallowing. The relationship between bolus driving pressures and residue remaining in the valleculae after the swallow has not been defined. Thirty-seven dysphagic patients who were evaluated with both videofluoroscopy (VFSS) and high-resolution manometry (HRM) were identified within the University of Wisconsin Voice and Swallowing Outcomes database. Patients were categorized according to binary ratings of presence or absence of vallecular stasis as well as incomplete or complete tongue retraction on VFSS. Tongue base region pressures measured with HRM during saline swallows of 1 and 10 ml volumes were compared to ratings of vallecular stasis or tongue base retraction. No significant difference could be identified among mean peak HRM pressures when compared to presence or absence of vallecular stasis (1 ml saline: p = .1886; 10 ml saline: p = .7354). When categorized according to complete or incomplete tongue retraction, mean peak HRM pressures were significantly greater in the complete tongue retraction group as compared to incomplete tongue retraction (1 ml saline: p = .0223; 10 ml saline: p = .0100). Findings suggest there are multiple factors that lead to reduced vallecular clearance. In the absence of HRM measures, judging complete or incomplete tongue retraction on VFSS may be a more valid gauge of tongue base region pressures than vallecular clearance when planning dysphagia treatment.


Fragile X-associated tremor/ataxia phenotype in a male carrier of unmethylated full mutation in the FMR1 gene.
The Fragile X syndrome is caused by a CGG repeat expansion >200 in the promoter of the Fragile X mental retardation 1 (FMR1) gene termed full mutation (FM). These alleles are silenced through methylation of the FMR1 promoter, leading to deficit of the FMR1 protein (FMRP), and neurodevelopmental changes. However, occasional FM individuals have a complete lack of methylation, and those typically have only minor deficit of FMRP levels compared with normal controls and their intelligence may be in the normal range. FM alleles are generated through expansion of the CGG repeat from the premutation (PM) range of 55-200 repeats, linked to the late onset Fragile X-associated tremor/ataxia syndrome (FXTAS). This disorder has been attributed to a 'toxicity' of the FMR1 mRNA, which is significantly elevated in male carriers of PM alleles and of unmethylated FM alleles. This is the first report of a 65-year-old male with an unmethylated FM allele and history of alcohol abuse, who developed symptoms of FXTAS. We postulate that, although the elevation of FMR1 transcripts associated with unmethylated FM alleles have a potential to cause FXTAS, in some cases this disorder may occur through an additional effect of exposure to neurotoxicants including alcohol.


Isolation of a new β-carboline alkaloid from aerial parts of Triclisia sacleuxii and its antibacterial and cytotoxicity effects.
A new β-carboline alkaloid named sacleuximine A (1) together with known compounds palmatine (2), isotetrandrine (3), trans-N-feruloyltyramine (4), trans-N-caffeoyltyramine (5), yangambin (6), syringaresinol (7), sesamin (8), (+) epi-quercitol (9), 4-hydroxybenzaldehyde (10), β-sitosterol (11), quercetin 3-O-rutinoside (12) and myricetin 3-O-β-glucose (1→6) α-rhamnoside (13) have been isolated from methanol extract of Triclisia sacleuxii aerial parts. Compounds 1-10 were evaluated for their cytotoxicity against human adenocarcinoma (HeLa), human hepatocarcinoma (Hep3B) and human breast carcinoma (MCF-7) cells lines and also for antibacterial activities against both Gram-positive and Gram-negative bacteria. The cytotoxicity (IC50) values ranged between 0.15 and 36.7 μM while the minimum inhibitory concentrations were found to be in the range of 3.9 and 125 μM, respectively. This is the first report of antibacterial compounds and the isolation of lignans together with a β-carboline alkaloid from T. sacleuxii.


Seasonal variation in the role of grey squirrels as hosts of Ixodes ricinus, the tick vector of the Lyme disease spirochaete, in a British woodland.
Data are presented on the variable patterns of the seasonal dynamics of Ixodes ricinus L. ticks seen questing on the vegetation and feeding on small rodents (mice and voles) and squirrels within a British woodland focus of Lyme borreliosis. Information on tick infestation levels on pheasants is also presented. The results show a prolonged, unimodal pattern of tick activity, with ticks feeding throughout the year in this sheltered habitat. If host density is taken into account, squirrels are quantitatively more important than small mammals as hosts for larval ticks from April until July, and overwhelmingly so for nymphal ticks throughout the year. The observed inter- and intraspecific differences in tick infestation levels are related to the behaviour of both hosts and ticks. Squirrels, as competent hosts for Borrelia burgdorferi and frequent occupants of habitats closely associated with man, will contribute significantly to the risk of Lyme disease.


fMRI assessment of upper extremity related brain activation with an MRI-compatible manipulandum.
Longitudinal studies to evaluate the effect of rehabilitative therapies require an objective, reproducible and quantitative means for testing function in vivo. An fMRI assessment tool for upper extremity related brain activation using an MRI-compatible manipulandum was developed and tested for use in neurorehabilitation research. Fifteen healthy, right-handed subjects participated in two fMRI sessions, which were three to four weeks apart. A block design paradigm, composed of three conditions of subject-passive movement, subject-active movement and rest, was employed for the fMRI recordings. During the rest condition, subjects simply held the device handle without applying any force or movement. The same type of auditory and visual instructions were given in all the three conditions, guiding the subjects to perform the motor tasks interactively with the MRI-compatible arm manipulandum. The tasks were controlled across the fMRI sessions. The subjects' brain activation was recorded by fMRI, and their behavioral performance was recorded by the manipulandum. The brain network activated by the subjects' interaction with the manipulandum was identified, and the reproducibility and reliability of the obtained activation were determined. All subjects completed the trial protocol. Two subjects were excluded from analysis due to head motion artifacts. All passive movements were performed well. Four out of the total 780 active movements were missed by two subjects. Brain activation was found in the contralateral sensorimotor cortex, secondary somatosensory cortex and non-primary motor cortex as well as in subcortical areas in the thalamus, basal ganglia and the cerebellum. These activations were consistent across the two fMRI sessions. The MRI-compatible manipulandum elicited robust and reproducible brain activations in healthy subjects during the subject-active and subject-passive upper extremity motor tasks with a block design paradigm. This system is promising for many applications in neurorehabilitation research and may be useful for longitudinal studies.


Association between two mu-opioid receptor gene (OPRM1) haplotype blocks and drug or alcohol dependence.
We examined 13 single nucleotide polymorphisms (SNPs) spanning the coding region of the mu-opioid receptor gene (OPRM1), among 382 European Americans (EAs) affected with substance dependence [alcohol dependence (AD) and/or drug dependence (DD)] and 338 EA healthy controls. These SNPs delineated two haplotype blocks. Genotype distributions for all SNPs were in Hardy-Weinberg equilibrium (HWE) in controls, but in cases, four SNPs in Block I and three SNPs in Block II showed deviation from HWE. Significant differences were found between cases and controls in allele and/or genotype frequencies for six SNPs in Block I and two SNPs in Block II. Association of SNP4 in Block I with DD (allele: P=0.004), SNP5 in Block I with AD and DD (allele: P< or =0.005 for both) and two SNPs in Block II with AD (SNP11 genotype: P=0.002; SNP12 genotype: P=0.001) were significant after correction for multiple testing. Frequency distributions of haplotypes (constructed by five tag SNPs) differed significantly for cases and controls (P<0.001 for both AD and DD). Logistic regression analyses confirmed the association between OPRM1 variants and substance dependence, when sex and age of subjects and alleles, genotypes, haplotypes or diplotypes of five tag SNPs were considered. Population structure analyses excluded population stratification artifact. Additional supporting evidence for association between OPRM1 and AD was obtained in a smaller Russian sample (247 cases and 100 controls). These findings suggest that OPRM1 intronic variants play a role in susceptibility to AD and DD in populations of European ancestry.


Skin cancer in heart transplant recipients.
The frequency of skin cancer in organ transplant recipients is high, up to 15%. Our purpose was to determine the incidence of skin cancer in patients who underwent immunosuppression after heart transplantation and to determine the factors important in the appearance of skin cancer. We studied the frequency of skin cancer in 92 of 111 patients after they underwent heart transplantation between January 1984 and December 1993. At least one cutaneous neoplasm (squamous cell carcinoma and/or basal cell carcinoma) developed in 14 patients (15.2%). The basal cell carcinoma to squamous cell carcinoma ratio was 1:1.5. The skin cancer appeared an average of 31.5 months after transplantation; the average was 36 months for squamous cell carcinoma and 25.3 months for basal cell carcinoma. Cumulative risk rose from 4.3% at 1 year up to 43.8% at 7 years after transplantation. The overall incidence of both types of skin cancer was 45.3 per 1000 posttransplant person-years, with an incidence of 25.8 for basal cell carcinoma and 29.1 for squamous cell carcinoma. Most skin cancers developed between 2 and 3 years after transplantation. All patients were exposed to a significant amount of ultraviolet radiation and had skin type II or III. We did not find a significant association between skin cancer and haplotype HLA-A3, HLA-A11, HLA-DR, and the number of mismatches for HLA-B. We found an increased progressive cumulative incidence of skin cancer in heart transplant recipients for two reasons: (1) immunosuppression and increased exposure to ultraviolet radiation in some patients, and (2) the skin type of certain patients. We emphasize the need for photoprotection in this group of patients and regular skin cancer screening examinations.


The effectiveness of intermediate care in a nursing-led in-patient unit.
In order to assess the potential for a nursing-led in-patient unit (NLIU) to substitute for a period of care in the acute hospital environment and promote recovery before discharge, a randomised controlled trial was conducted. The setting was an acute inner London hospital trust, part of the UK's national health service. Of patients referred to a NLIU from acute wards, 80 were randomly assigned to usual care (remain in normal hospital system) and 97 to the NLIU (nursing-led care with no routine medical involvement). Patients were identified as medically stable but in need of additional nursing intervention by referring medical staff prior to full nursing assessment of suitability. Outcomes compared included functional dependence (Barthel Index), discharge destination and length of hospital stay. Inputs from nursing, paramedical and medical staff were measured. There was no significant difference in functional independence at discharge (p0.05). Patients undergoing usual care stayed in hospital for less time (mean difference 18 days, p<0.01) but the same number of patients were in hospital 90 days after recruitment (23% NLIU, 24% usual care p0.05) due to re-admissions. The model of care implemented differed considerably from that described in the literature with the NLIU having significantly fewer qualified nurses (RNs). Although the anticipated benefits of the NLIU were not demonstrated, the study does not conclude that the model should be rejected. Factors driving length of stay need to be further investigated, as does the possibility of post-discharge benefits. The NLIU does offer some potential to substitute for acute care but also appears to substitute for a period of primary care.


Zinc transport and the inhibition of the L-type calcium channel are two separable functions of ZnT-1.
Traditionally, proteins are considered to perform a single role, be it as an enzyme, a channel, a transporter or as a structural scaffold. However, recent studies have described moonlighting proteins that perform distinct and independent functions; for example, TRPM7 is both an ion channel and a kinase. ZnT-1 is a member of the Carrier Diffusion Facilitator family that is expressed throughout the phylogenetic tree from bacteria to humans. Since its cloning in 1995, ZnT-1 is considered a major extruder of Zn2+ based on its capability to protect cells against zinc toxicity. Recently, we reported that ZnT-1 inhibits the L-type calcium channel (LTCC), a major Zn2+ and Ca2+ entry pathway. Here we show that ZnT-1 is a dual-function protein by demonstrating that its abilities to exchange Zn2+/H+ and to inhibit the LTCC are independent of each other and are mediated by different parts of the protein. Specifically, mutations in the membrane-spanning helices that render ZnT-1 unable to transport zinc do not prevent it from inhibiting the LTCC. Moreover, a fragment consisting of the intracellular ZnT-1 C-terminal, which lacks all ion-transfer segments, inhibits the LTCC as efficiently as wild-type ZnT-1. Our data therefore indicates that ZnT-1 performs two structurally independent functions related to zinc homeostasis.


Influence of lead apron shielding on absorbed doses from panoramic radiography.
This study investigated the absorbed doses in a full anthropomorphic body phantom from two different panoramic radiography devices, performing protocols with and without applying a lead apron. A RANDO® full body phantom (Alderson Research Laboratories Inc., Stamford, CT) was equipped with 110 thermoluminescent dosemeters at 55 different sites and set up in two different panoramic radiography devices [SCANORA® three-dimensional (3D) (SOREDEX, Tuusula, Finland) and ProMax® 3D (Planmeca, Helsinki, Finland)] and exposed. Two different protocols were performed in the two devices. The first protocol was performed without any lead shielding, whereas the phantom was equipped with a standard adult lead apron for the second protocol. A two-tailed paired samples t-test for the SCANORA 3D revealed that there is no difference between the protocol using lead apron shielding (m = 87.99, s = 102.98) and the protocol without shielding (m = 87.34, s = 107.49), t(54) = −0.313, p > 0.05. The same test for the ProMax 3D showed that there is also no difference between the protocol using shielding (m = 106.48, s = 117.38) and the protocol without shielding (m = 107.75, s = 114,36), t(54) = 0.938, p > 0.05. In conclusion, the results of this study showed no statistically significant differences between a panoramic radiography with or without the use of lead apron shielding.


Thiamin and Riboflavin in Human Milk: Effects of Lipid-Based Nutrient Supplementation and Stage of Lactation on Vitamer Secretion and Contributions to Total Vitamin Content.
While thiamin and riboflavin in breast milk have been analyzed for over 50 years, less attention has been given to the different forms of each vitamin. Thiamin-monophosphate (TMP) and free thiamin contribute to total thiamin content; flavin adenine-dinucleotide (FAD) and free riboflavin are the main contributors to total riboflavin. We analyzed milk collected at 2 (n = 258) or 6 (n = 104), and 24 weeks (n = 362) from HIV-infected Malawian mothers within the Breastfeeding, Antiretrovirals and Nutrition (BAN) study, randomly assigned at delivery to lipid-based nutrient supplements (LNS) or a control group, to investigate each vitamer's contribution to total milk vitamin content and the effects of supplementation on the different thiamin and riboflavin vitamers at early and later stages of lactation, and obtain insight into the transport and distribution of these vitamers in human milk. Thiamin vitamers were derivatized into thiochrome-esters and analyzed by high-performance liquid-chromatography-fluorescence-detection (HPLC-FLD). Riboflavin and FAD were analyzed by ultra-performance liquid-chromatography-tandem-mass-spectrometry (ULPC-MS/MS). Thiamin-pyrophosphate (TPP), identified here for the first time in breast milk, contributed 1.9-4.5% to total thiamin. Free thiamin increased significantly from 2/6 to 24 weeks regardless of treatment indicating an active transport of this vitamer in milk. LNS significantly increased TMP and free thiamin only at 2 weeks compared to the control: median 170 versus 151 μg/L (TMP), 13.3 versus 10.5 μg/L (free thiamin, p<0.05 for both, suggesting an up-regulated active mechanism for TMP and free thiamin accumulation at early stages of lactation. Free riboflavin was consistently and significantly increased with LNS (range: 14.8-19.6 μg/L (LNS) versus 5.0-7.4 μg/L (control), p<0.001), shifting FAD:riboflavin relative amounts from 92-94:6-8% to 85:15%, indicating a preferred secretion of the free form into breast milk. The continuous presence of FAD in breast milk suggests an active transport and secretion system for this vitamer or possibly formation of this co-enymatic form in the mammary gland.


Experiences, Expectations, and Satisfaction with Police-a Population-Based Study of Differences by Race and Ethnicity in Two Counties.
We used the 2021 Policing in America Survey to examine experiences with police and perceptions of policing in Cook County, IL, and Dallas County, TX. Substantial portions of residents believe that local police use force during at least half of arrests (Cook 45.8%, Dallas 52.2%) and report dissatisfaction with local policing efforts (Cook 49.0%, Dallas 48.5%). Black and Hispanic residents in both communities were more likely than white, non-Hispanic residents to have frequent use of force expectations and dissatisfaction with police. Several other indicators of experiences with and perceptions of police also varied by race and ethnicity. We combined individual-level survey data with neighborhood-level data from the American Community Survey in multilevel logistic regression models to examine differences between groups for primary outcomes. Some findings were consistent for both counties. Black residents (Cook aOR = 3.63; Dallas aOR = 5.07) and those who had witnessed police misconduct were more likely to have frequent use of force expectations (Cook aOR = 6.44; Dallas aOR = 14.91). Differences between Black, Hispanic, and white residents were not associated with dissatisfaction in the adjusted models. Negative experiences with police (Cook aOR = 10.73; Dallas aOR = 12.11), witnessing misconduct (Cook aOR = 3.87; Dallas aOR = 3.55), and feeling unsafe in one's neighborhood (Cook aOR = 4.56; Dallas aOR = 3.49) were independently associated with greater odds of dissatisfaction. Inequities in exposure to unsafe neighborhoods, witnessing misconduct, and negative interactions with police are key drivers of variation in expectations and perceptions of police. Optimizing transparency and assurances of procedural justice are needed to reduce fear and increase satisfaction with police in the USA.


Human regulatory factor X 4 (RFX4) is a testis-specific dimeric DNA-binding protein that cooperates with other human RFX members.
Regulatory factor X (RFX) members are evolutionarily conserved transcription factors that share a highly conserved winged helix DNA-binding domain. Human RFX4 has been isolated from breast cancer as a partial cDNA encoding a short RFX-type DNA-binding domain fused to the estrogen receptor, but the entire structure of RFX4 has been unknown. Here, we report the molecular cloning and characterization of human RFX4. RFX4 contains evolutionarily conserved regions, including a RFX-type DNA-binding domain, a dimerization domain, and other conserved regions, and is closely related to RFX1, RFX2, and RFX3 in structure. The expression of RFX4 is restricted to testis. In vitro synthesized RFX4 protein bound to typical RFX binding sites in a sequence-dependent manner. Immunoprecipitation analyses showed that RFX4 interacts physically with RFX2, RFX3, and RFX4 itself but not with RFX1. In contrast to other mammalian RFX members that form dimers, RFX4 is revealed to have no distinct transcriptional activation domains. By using a chimeric protein of RFX1 and RFX4, the C-terminal domain of RFX4 was shown to be a possible transcriptional repression domain. Taken together, these results indicate that RFX4 is the first mammalian member of RFX family without transcriptional activation capacity and might function through selective interactions with other RFX members in transcriptional regulation.


Pediatric Outcomes Data Collection Instrument is a Useful Patient-Reported Outcome Measure for Physical Function in Children with Osteogenesis Imperfecta.
Patient-reported outcome measures (PROMs) are increasingly recognized as valuable endpoints in clinical trials. The Pediatric Outcomes Data Collection Instrument (PODCI) is a PROM utilized in children with musculoskeletal disorders. We evaluated the validity and reliability of PODCI in children with osteogenesis imperfecta (OI). Physical functioning and psychological well-being were assessed using PODCI in a large cohort of children enrolled in a multicenter study conducted by the Brittle Bone Disorders Consortium. Physical function scores were correlated with a validated, observer-rated scale, Brief Assessment of Motor Function (BAMF), and with psychological well-being scores. We calculated sample sizes required to detect clinically meaningful differences in physical function. Four hundred seventeen children with OI types I, III, and IV were enrolled. Physical function scores in OI type III were significantly lower than those in OI types I and IV. There were no significant differences in psychological well-being. PODCI physical function scores showed moderate-to-strong correlation with BAMF. The Global Functioning Scale, a composite of physical function, did not consistently correlate with psychological well-being. PODCI can be a reliable measure of physical functioning in children with OI and offers valuable information about patient-reported health status and new ways to examine the utility of interventions in this population.


Multiple-wavelength surface patterns in models of biological chiral liquid crystal membranes.
We present a model to investigate the formation of surface patterns in biological materials through the interaction of anisotropic interfacial tension, bending elasticity, and capillarity at their free surfaces. Focusing on the cholesteric liquid crystal (CLC) material model, the generalized shape equation for anisotropic interfaces using the Rapini-Papoular anchoring and Helfrich free energies is applied to understand the formation of multi-length scale patterns, such as those found in floral petals. The chiral liquid crystal-membrane model is shown to be analogous to a driven pendulum, a connection that enables generic pattern classification as a function of bending elasticity, liquid crystal chirality and anchoring strength. The unique pattern-formation mechanism emerging from the model here presented is based on the nonlinear interaction between bending-driven folding and anchoring-driven creasing. The predictions are shown to capture accurately the two-scale wrinkling of certain tulips. These new findings enable not only to establish a new paradigm for characterizing surface wrinkling in biological liquid crystals, but also to inspire the design of functional surface structures.


Repressor element 1-silencing transcription factor deficiency yields profound hearing loss through Kv7.4 channel upsurge in auditory neurons and hair cells.
Repressor element 1-silencing transcription factor (REST) is a transcriptional repressor that recognizes neuron-restrictive silencer elements in the mammalian genomes in a tissue- and cell-specific manner. The identity of REST target genes and molecular details of how REST regulates them are emerging. We performed conditional null deletion of Rest (cKO), mainly restricted to murine hair cells (HCs) and auditory neurons (aka spiral ganglion neurons [SGNs]). Null inactivation of full-length REST did not affect the development of normal HCs and SGNs but manifested as progressive hearing loss in adult mice. We found that the inactivation of REST resulted in an increased abundance of Kv7.4 channels at the transcript, protein, and functional levels. Specifically, we found that SGNs and HCs from Rest cKO mice displayed increased Kv7.4 expression and augmented Kv7 currents; SGN's excitability was also significantly reduced. Administration of a compound with Kv7.4 channel activator activity, fasudil, recapitulated progressive hearing loss in mice. In contrast, inhibition of the Kv7 channels by XE991 rescued the auditory phenotype of Rest cKO mice. Previous studies identified some loss-of-function mutations within the Kv7.4-coding gene, Kcnq4, as a causative factor for progressive hearing loss in mice and humans. Thus, the findings reveal that a critical homeostatic Kv7.4 channel level is required for proper auditory functions.


Airflow pattern complexity and airway obstruction in asthma.
The scientific and clinical value of a measure of complexity is potentially enormous because complexity appears to be lost in the presence of illness. The authors examined the effect of elevated airway obstruction on the complexity of the airflow (Q) pattern of asthmatic patients analyzing the airflow approximate entropy (ApEnQ). This study involved 11 healthy controls, 11 asthmatics with normal spirometric exams, and 40 asthmatics with mild (14), moderate (14), and severe (12) airway obstructions. A significant (P < 0.02) reduction in the ApEnQ was observed in the asthmatic patients. This reduction was significantly correlated with spirometric indexes of airway obstruction [FEV(1) (%): R = 0.31, P = 0.013] and the total respiratory impedance (R = -0.39; P < 0.002). These results are in close agreement with pathophysiological fundamentals and suggest that the airflow pattern becomes less complex in asthmatic patients, which may reduce the adaptability of the respiratory system to perform the exercise that is associated with daily life activities. This analysis was able to identify respiratory changes in patients with mild obstruction with an adequate accuracy (83%). Higher accuracies were obtained in patients with moderate and severe obstructions. The analysis of airflow pattern complexity by the ApEnQ was able to provide new information concerning the changes associated with asthma. In addition, this analysis was also able to contribute to the detection of the adverse effects of asthma. Because these measurements are easy to perform, such a technique may represent an alternative and/or a complement to other conventional exams to help the clinical evaluations of asthmatic patients.


Executive function predicts cognitive-behavioral therapy response in childhood obsessive-compulsive disorder.
Cognitive-behavioral therapy (CBT) is considered first-line treatment for childhood obsessive-compulsive disorder (OCD). Despite CBT's efficacy, too many children and adolescents do not fully respond to treatment, making the identification of predictors of treatment response highly relevant. Executive functions (EF) have been suggested to constitute such predictors, but studies with pediatric samples are scarce. In the present study, we investigated latent level EF test performance and ratings of daily life EF behavior as predictors of CBT response in pediatric OCD. We further examined the stability of EF from pre-to post-treatment and the association of EF changes with OCD severity change. EF test performance significantly predicted exposure-based CBT outcome. Patients with better EF test performance had significantly elevated risk of non-response relative to patients with poorer performance. Daily life EF behavior in OCD probands improved after treatment relative to controls. The findings suggest that EF performance impacts CBT outcome, and that exposure-based CBT is well-suited for children and adolescents with OCD and poorer EF test performance. This study supports the relevance of EF in CBT for childhood OCD and denotes a possible need for development of enhanced treatments for children and adolescents with OCD and superior EF performance.


Regulation of VEGF-induced endothelial cell migration by mitochondrial reactive oxygen species.
Endothelial migration is a crucial aspect of a variety of physiologic and pathologic conditions including atherosclerosis and vascular repair. Reactive oxygen species (ROS) function as second messengers during endothelial migration. Multiple intracellular sources of ROS are regulated by cellular context, external stimulus, and the microenvironment. However, the predominant source of ROS during endothelial cell (EC) migration and the mechanisms by which ROS regulate cell migration are incompletely understood. In this study, we tested the hypothesis that mitochondria-derived ROS (mtROS) regulate EC migration. In cultured human umbilical vein endothelial cells, VEGF increased mitochondrial metabolism, promoted mtROS production, and induced cell migration. Either the targeted mitochondrial delivery of the antioxidant, vitamin E (Mito-Vit-E), or the depletion of mitochondrial DNA abrogated VEGF-mediated mtROS production. Overexpression of mitochondrial catalase also inhibited VEGF-induced mitochondrial metabolism, Rac activation, and cell migration. Furthermore, these interventions suppressed VEGF-stimulated EC migration and blocked Rac1 activation in endothelial cells. Constitutively active Rac1 reversed Mito-Vit-E-induced inhibition of EC migration. Mito-Vit-E also attenuated carotid artery reendothelialization in vivo. These results provide strong evidence that mtROS regulate EC migration through Rac-1.


Redox proteomics in selected neurodegenerative disorders: from its infancy to future applications.
Several studies demonstrated that oxidative damage is a characteristic feature of many neurodegenerative diseases. The accumulation of oxidatively modified proteins may disrupt cellular functions by affecting protein expression, protein turnover, cell signaling, and induction of apoptosis and necrosis, suggesting that protein oxidation could have both physiological and pathological significance. For nearly two decades, our laboratory focused particular attention on studying oxidative damage of proteins and how their chemical modifications induced by reactive oxygen species/reactive nitrogen species correlate with pathology, biochemical alterations, and clinical presentations of Alzheimer's disease. This comprehensive article outlines basic knowledge of oxidative modification of proteins and lipids, followed by the principles of redox proteomics analysis, which also involve recent advances of mass spectrometry technology, and its application to selected age-related neurodegenerative diseases. Redox proteomics results obtained in different diseases and animal models thereof may provide new insights into the main mechanisms involved in the pathogenesis and progression of oxidative-stress-related neurodegenerative disorders. Redox proteomics can be considered a multifaceted approach that has the potential to provide insights into the molecular mechanisms of a disease, to find disease markers, as well as to identify potential targets for drug therapy. Considering the importance of a better understanding of the cause/effect of protein dysfunction in the pathogenesis and progression of neurodegenerative disorders, this article provides an overview of the intrinsic power of the redox proteomics approach together with the most significant results obtained by our laboratory and others during almost 10 years of research on neurodegenerative disorders since we initiated the field of redox proteomics.


[Experimental study on gut tolerance to early enteral nutrition following intestinal ischemia/reperfusion].
To investigate the effects of early enteral nutrition (EEN) on intestinal function and gut tolerance after intestinal ischemia/reperfusion (I/R) injury. Twenty-four male dogs were randomly divided into three groups: EEN only, I/R only and I/R+EEN. The superior mesenteric artery (SMA) was blocked for 1 hour followed by restoration of blood flow. EEN was given 4 hours after reperfusion by continuous infusion of Fresubin nutrition fluid (4 ml.kg(-1).h(-1), Fresenius Kabi Co, Germany) into gut via a tonometric catheter for 3 hours till intolerance symptoms, which including vomiting and diarrhea which indicated gut intolerance. CO(2) partial pressure of intestinal mucosa (PiCO(2)), D-xylose absorption and intestinal luminal pressure were measured to reflect intestinal function and perfusion. Incidence and severity of diarrhea and vomiting were significantly higher in I/R+EEN group (with 87.5% intolerance) than those in I/R only group (12.5%) and EEN only group (0). After EEN, PiCO(2) and intestinal cavity pressure were significantly higher, and the D-xylose absorption much lower, in the I/R+EEN group compared with the I/R group and EEN group (all P<0.01). Intestinal I/R may result in decreased tolerance to EEN. Too early enteral nutrition (less than 12 hours after gut hypoperfusion) may enhance intestinal ischemia injury and further inhibit its function of propulsion and absorption.


Evaluation of the Dutch version of the Parkinson's Disease Questionnaire 39.
The psychometric properties of the Dutch version of the Parkinson's disease questionnaire 39 (PDQ39-DV) were tested in 177 patients with Parkinson's disease (PD). Internal consistency of 7 of the 8 scales was adequate (>or=0.70), but was insufficient for 'bodily discomfort' (0.59). Correlation between the PDQ39 and other instruments in this study, the SCOPA-psychosocial questionnaire, Euroqol-5D, hospital anxiety and depression scale, and a visual analogue scale for quality of life, were 0.82, 0.74, 0.63 and -0.54, respectively. The factor analysis yielded 8 factors, which accounted for 65% of the variance and revealed only small differences with the original UK version. We conclude that the PDQ39-DV displays satisfactory psychometric properties and is an appropriate instrument to assess quality of life in Dutch patients with PD.


Processing Disability.
This Article argues that the practice of holding so many adjudicative proceedings related to disability in private settings (e.g., guardianship, special education due process, civil commitment, and social security) relative to our strong normative presumption of public access to adjudication may cultivate and perpetuate stigma in contravention of the goals of inclusion and enhanced agency set forth in antidiscrimination laws. Descriptively, the law has a complicated history with disability--initially rendering disability invisible; later, underwriting particular narratives of disability synonymous with incapacity; and, in recent history, promoting the full socio-economic visibility of people with disabilities. The Americans with Disabilities Act (ADA), the marquee civil rights legislation for people with disabilities (about to enter its twenty-fifth year), expresses a national approach to disability that recognizes the role of society in its construction, maintenance, and potential remedy. However, the ADA's mission is incomplete. It has not generated the types of interactions between people with disabilities and nondisabled people empirically shown to deconstruct deeply entrenched social stigma. Prescriptively, procedural design can act as an "ntistigma agent"to resist and mitigate disability stigma. This Article focuses on one element of institutional design--public access to adjudication--as a potential tool to construct and disseminate counter-narratives of disability. The unique substantive focus in disability adjudication on questions of agency provides a potential public space for the negotiation of nuanced definitions of disability and capacity more reflective of the human condition.


Decomposing complex reaction networks using random sampling, principal component analysis and basis rotation.
Metabolism and its regulation constitute a large fraction of the molecular activity within cells. The control of cellular metabolic state is mediated by numerous molecular mechanisms, which in effect position the metabolic network flux state at specific locations within a mathematically-definable steady-state flux space. Post-translational regulation constitutes a large class of these mechanisms, and decades of research indicate that achieving a network flux state through post-translational metabolic regulation is both a complex and complicated regulatory problem. No analysis method for the objective, top-down assessment of such regulation problems in large biochemical networks has been presented and demonstrated. We show that the use of Monte Carlo sampling of the steady-state flux space of a cell-scale metabolic system in conjunction with Principal Component Analysis and eigenvector rotation results in a low-dimensional and biochemically interpretable decomposition of the steady flux states of the system. This decomposition comes in the form of a low number of small reaction sets whose flux variability accounts for nearly all of the flux variability in the entire system. This result indicates an underlying simplicity and implies that the regulation of a relatively low number of reaction sets can essentially determine the flux state of the entire network in the given growth environment. We demonstrate how our top-down analysis of networks can be used to determine key regulatory requirements independent of specific parameters and mechanisms. Our approach complements the reductionist approach to elucidation of regulatory mechanisms and facilitates the development of our understanding of global regulatory strategies in biological networks.


Spectroscopic study of a DNA brush synthesized in situ by surface initiated enzymatic polymerization.
We used a combination of synchrotron-based X-ray photoelectron spectroscopy (XPS) and angle-resolved near-edge X-ray absorption fine structure (NEXAFS) spectroscopy to study the chemical integrity, purity, and possible internal alignment of single-strand (ss) adenine deoxynucleotide (poly(A)) DNA brushes. The brushes were synthesized by surface-initiated enzymatic polymerization (SIEP) on a 25-mer of adenine self-assembled monolayer (SAM) on gold (A25-SH), wherein the terminal 3'-OH of the A25-SH serve as the initiation sites for SIEP of poly(A). XPS and NEXAFS spectra of poly(A) brushes were found to be almost identical to those of A25-SH initiator, with no unambiguous traces of contamination. Apart from the well-defined chemical integrity and contamination-free character, the brushes were found to have a high degree of orientational order, with an upright orientation of individual strands, despite their large thickness up to ~55 nm, that corresponds to a chain length of at least several hundred nucleotides for individual ssDNA molecules. The orientational order exhibited by these poly(A) DNA brushes, mediated presumably by base stacking, was found to be independent of the brush thickness as long as the packing density was high enough. The well-defined character and orientational ordering of the ssDNA brushes make them a potentially promising system for different applications.


Uncertainty Quantification of Oscillation Suppression During DBS in a Coupled Finite Element and Network Model.
Models of the cortico-basal ganglia network and volume conductor models of the brain can provide insight into the mechanisms of action of deep brain stimulation (DBS). In this study, the coupling of a network model, under parkinsonian conditions, to the extracellular field distribution obtained from a three dimensional finite element model of a rodent's brain during DBS is presented. This coupled model is used to investigate the influence of uncertainty in the electrical properties of brain tissue and encapsulation tissue, formed around the electrode after implantation, on the suppression of oscillatory neural activity during DBS. The resulting uncertainty in this effect of DBS on the network activity is quantified using a computationally efficient and non-intrusive stochastic approach based on the generalized Polynomial Chaos. The results suggest that variations in the electrical properties of brain tissue may have a substantial influence on the level of suppression of oscillatory activity during DBS. Applying a global sensitivity analysis on the suppression of the simulated oscillatory activity showed that the influence of uncertainty in the electrical properties of the encapsulation tissue had only a minor influence, in agreement with previous experimental and computational studies investigating the mechanisms of current-controlled DBS in the literature.


A UK survey of virological testing of cadaver tissue donors. Microbiology Working Group of the NIBSC steering group on Tissue/Cell Banking and Engineering.
To analyse information about virological testing of cadaveric tissue donors, including the kits used and the rates of test reactivity. Retrospective data were collected using a standardised questionnaire sent to 16 tissue banks in the UK. The rates of repeat reactive screen tests and confirmed positive results for the markers HBsAg, anti-HCV, anti-HIV were analysed in 1,833 cadaver tissue donors tested at 12 tissue banks over one year up to March 1998. There was a wide range of kits in use for virological screen testing of cadaver donors. The rates of repeat reactivity in the screen tests varied from 0 to 42%. The findings have implications for policies based on discard of tissues from donors with repeat reactive results, and raise important safety issues with regards to cadaveric virological testing, as the test systems in use have not been validated for cadaver blood samples.


Surprising solvent-induced structural rearrangements in large [N···I+···N] halogen-bonded supramolecular capsules: an ion mobility-mass spectrometry study.
Coordinative halogen bonds have recently gained interest for the assembly of supramolecular capsules. Ion mobility-mass spectrometry and theoretical calculations now reveal the well-defined gas-phase structures of dimeric and hexameric [N···I+···N] halogen-bonded capsules with counterions located inside their cavities as guests. The solution reactivity of the large hexameric capsule shows the intriguing solvent-dependent equilibrium between the hexamer and an unprecedented pentameric [N···I+···N] halogen-bonded capsule, when the solvent is changed from chloroform to dichloromethane. The intrinsic flexibility of the cavitands enables this novel structure to adopt a pseudo-trigonal bipyramidal geometry with nine [N···I+···N] bonds along the edges and two pyridine binding sites uncomplexed.


Size-class effect contributes to tree species assembly through influencing dispersal in tropical forests.
We have investigated the processes of community assembly using size classes of trees. Specifically our work examined (1) whether point process models incorporating an effect of size-class produce more realistic summary outcomes than do models without this effect; (2) which of three selected models incorporating, respectively environmental effects, dispersal and the joint-effect of both of these, is most useful in explaining species-area relationships (SARs) and point dispersion patterns. For this evaluation we used tree species data from the 50-ha forest dynamics plot in Barro Colorado Island, Panama and the comparable 20 ha plot at Bubeng, Southwest China. Our results demonstrated that incorporating an size-class effect dramatically improved the SAR estimation at both the plots when the dispersal only model was used. The joint effect model produced similar improvement but only for the 50-ha plot in Panama. The point patterns results were not improved by incorporation of size-class effects using any of the three models. Our results indicate that dispersal is likely to be a key process determining both SARs and point patterns. The environment-only model and joint-effects model were effective at the species level and the community level, respectively. We conclude that it is critical to use multiple summary characteristics when modelling spatial patterns at the species and community levels if a comprehensive understanding of the ecological processes that shape species' distributions is sought; without this results may have inherent biases. By influencing dispersal, the effect of size-class contributes to species assembly and enhances our understanding of species coexistence.


The struggle for sanitary reform in the Lancashire cotton mills, 1920-1970.
This paper reports one aspect of a larger study. The aim of this aspect was to explore the role of the "welfare officer" in promoting the health of cotton mill workers during this period. The paper considers one element of a broad exploratory study of the health of women cotton mill workers in the North West of England. The original purpose of the study, which was conducted in 2002 in two towns, Oldham and Ashton-under-Lyne, was to explore the women's own perceptions of the impact of their work on their health, and to find out what, if any, help and support they obtained. During the course of the study it was discovered that "welfare officers", some of whom were trained nurses, had an important role in promoting the health of these workers. The study was therefore expanded to incorporate data obtained directly from interviews with a small sample of welfare officers. The present paper focuses on the issue of sanitary reform and considers the role of the "welfare officer" in promoting public health in the workplace. The study employed a combination of archive searches and oral history interviewing. In total, 31 interviews were undertaken between June 2001 and October 2002. The interpretive process focussed both on the ideological power structures which influenced the perspectives of participants, and on evidence for those aspects of participants' experiences which impacted on their health. In considering their health, female cotton-mill workers recalled that the poor sanitary conditions in their workplaces during the middle years of the 20th century had been a source of some concern to them. They also observed that mill welfare officers took a hand in promoting improvement in available facilities. Welfare officers themselves recounted their own concern regarding the poor sanitary conditions in the mills, and their efforts to improve conditions for the mill workers. The paper demonstrates the role of a little-known group of health workers in the middle years of the 20th century, and demonstrates the importance of oral history work in re-capturing elements of nursing work and experience, which do not appear extensively in the written record. The study's relevance to contemporary practice lies in the insight it offers into the autonomy with which these occupational health workers defined their roles and performed their work.


Electron microscopic localization of the beta-adrenergic receptor using a ferritin-alprenolol probe.
This report describes electron microscopic localization of the beta-adrenergic receptor using a beta-adrenergic receptor antagonist conjugated to ferritin. The conjugate was synthesized by reacting a carboxylic acid derivative of alprenolol with ferritin. The ferritin-alprenolol compound was shown to be effective in displacing specific [3H]dihydroalprenolol binding from rat erythrocyte membranes with a dissociation constant (Kd) of 25 nM. Rat erythrocyte ghosts were incubated with the compound and quantitative electron micrographic analysis yielded total binding of 1367 +/- 129 ferritin particles and nonspecific binding of 688 +/- 111 (six experiments). Thus, specific binding was 680 +/- 60 ferritin particles per red cell profile. Qualitative observations suggested that the particles were distributed randomly on the surface of the erythrocyte, although an occasional cluster was seen. A compound from another synthesis was shown be to effective in displacing specific [125I]iodocyanopindolol binding from neonatal rat cardiac myocyte membranes, with a dissociation constant of 13.8 nM, whereas native alprenolol had a dissociation constant of 1.3 nM. Neonatal rat cardiac myocytes were incubated with the compound and processed for electron microscopy. Total binding along the sarcolemmal membrane was 504 +/- 38 ferritin particles/100 micron of membrane and nonspecific binding was 301 +/- 26 ferritin particles/100 micron of membrane (seven experiments), yielding specific binding of 203 ferritin particles/100 micron of membrane. In additional studies, specific binding was inhibited 95% with 10(-5) M l-isoproterenol and 29% with d-isoproterenol, indicating stereoselectivity (seven experiments). The probe was distributed randomly along the sarcolemma with no preferential localization to coated pits or other membrane specializations. From measurements of the surface area of the average cardiac myocyte (732 micron 2), the specific binding of ferritin-alprenolol per 100 micron of membrane (203), and section thickness (0.08 micron), we calculated that cardiac myocytes had 18,575 beta-adrenergic membrane receptor sites. Thus, we have described a method for synthesizing and applying an electron-dense probe for electron microscopic localization of beta-adrenergic receptors. In these studies we determined the distribution of these receptors on rat erythrocyte ghosts and neonatal rat cardiac myocytes.


Effects of cognitive function on gait and dual tasking abilities in patients with Parkinson's disease suffering from motor response fluctuations.
Recent studies have demonstrated that cognitive loading aggravates the gait impairments that are typically seen in Parkinson's disease (PD). To better understand the relationship between cognition and gait in PD, we evaluated 30 subjects with PD who suffer from motor response fluctuations. The subjects were clinically and cognitively assessed using standard clinical (e.g., Unified Parkinson's Disease Rating Scale) and cognitive tests while in the "ON" period of the medication cycle. In addition, the subjects wore force-sensitive insoles to quantify the timing of the gait cycles during 80-m walks at a self-selected, comfortable pace during three randomly presented gait conditions: (1) usual-walking, (2) dual tasking (DT), performing serial 3 subtractions (DT_S3), and (3) DT_S7. Stride length, gait speed, gait variability and bilateral coordination of gait were affected by DT, compared to the usual-walking (P < 0.001) as was gait asymmetry (P = 0.024). Stepwise regression analyses showed that a subset of the cognitive performance scores accounted for the changes seen in the gait parameters during DT, e.g., set shifting capabilities as expressed by the Trial Making Test Scores (P < 0.001). Affect (e.g., anxiety) was not associated with DT-related gait changes. For most gait features, DT had a large impact on the DT_S3 condition with only minimal additional effect in the DT_S7 condition. These results demonstrate that the complex cognitive-motor interplay in the control of gait in patients with PD who suffer from motor response fluctuations has a profound and marked effect during DT conditions on gait variability, asymmetry and bilateral coordination, even in the "ON" state when patients are likely to be most active, mobile and vulnerable to the negative effects of dual tasking.


Coupled tertiary folding and oligomerization of the T1 domain of Kv channels.
Acquisition of secondary, tertiary, and quaternary structure is critical to the fabrication, assembly, and function of ion channels, yet the relationship between these biogenic events remains unclear. We now address this issue in voltage-gated K(+) channels (Kv) for the T1 domain, an N-terminal Kv recognition domain that is responsible for subfamily-specific, efficient assembly of Kv subunits. This domain forms a 4-fold symmetric tetramer. We have identified residues along the axial T1-T1 interface that are critical for tertiary and quaternary structure, shown that mutations at one end of the axial T1 interface can perturb the crosslinking of an intersubunit cysteine pair at the other end, and demonstrated that tertiary folding and tetramerization of this Kv domain are coupled. A threshold level of tertiary folding is required for monomers to oligomerize. Coupling between tertiary and quaternary structure formation may be a common feature in the biogenesis of multimeric proteins.


An integrated material metabolism model for stocks of urban road system in Beijing, China.
Rapid urbanization has greatly altered the urban metabolism of material and energy. As a significant part of the infrastructure, urban roads are being rapidly developed worldwide. Quantitative analysis of metabolic processes on urban road systems, especially the scale, composition and spatial distribution of their stocks, could help to assess the resource appropriation and potential environmental impacts, as well as improve urban metabolism models. In this paper, an integrated model, which covered all types of roads, intersection structures and ancillary facilities, was built for calculating the material stocks of urban road systems. Based on a bottom-up method, the total stocks were disassembled into a number of stock parts rather than obtained by input-output data, which provided an approach promoting data availability and inner structure understanding. The combination with GIS enabled the model to tackle the complex structures of road networks and avoid double counting. In the case study of Beijing, the following results are shown: 1) The total stocks for the entire road system reached 159 million tons, of which nearly 80% was stored in roads, and 20% in ancillary facilities. 2) Macadam was the largest stock (111 million tons), while stone mastic asphalt, polyurethane plastics, and atactic polypropylene accounted for smaller components of the overall system. 3) The stock per unit area of pedestrian overcrossing was higher than that of the other stock units in the entire system, and its steel stocks reached 0.49 t/m(2), which was 10 times as high as that in interchanges. 4) The high stock areas were mainly distributed in ring-shaped and radial expressways, as well as in major interchanges. 5) Expressways and arterials were excessively emphasized, while minor roads were relatively ignored. However, the variation of cross-sectional thickness in branches and neighborhood roads will have a significant impact on the scale of material stocks in the entire road system.


Association of Helicobacter pylori infection with the metabolic syndrome among HIV-infected black Africans receiving highly active antiretroviral therapy.
The metabolic syndrome (MetS) is common in human immune deficiency virus (HIV)-infected individuals receiving highly active antiretroviral therapy (HAART). Immune deficiencies caused by HIV give rise to numerous opportunistic gastrointestinal pathogens such as Helicobacter pylori, the commonest cause of chronic gastritis. The study sought to determine the relationship between H pylori infection and the MetS among HIV-infected clinic attendees. This cross-sectional study was carried out in a specialised heart clinic in Kinshasa, DR Congo. Between January 2004 and December 2008, 116 HIV-infected patients (61 with MetS and 55 without MetS) who underwent upper gastrointestinal endoscopy for dyspeptic symptoms were included in the study following an informed consent. Univariate associations were determined by odds ratios (OR), while multivariate logistic regression analysis was used to identify factors associated with the MetS. H pylori infection (OR = 13.5, 95% CI: 10.3-17.6; p < 0.0001) and peripheral obesity (median hip circumference ≥ 97 cm) (OR = 4.7, 95% CI: 1.2-18.8; p = 0.029) were identified as MetS-related factors in HIV-infected patients. Higher rates of the MetS were associated with increased incidence of HIV-related immunocompromise using World Health Organisation (WHO) staging criteria. There was a univariate significant difference in the prevalence of the MetS between antiretroviral therapy (ART)-naïve patients and patients treated by means of a first-line HAART regimen of stavudine (d4T), lamivudine (3TC) and nevirapine (NVP). However, this difference was not significant in multivariate logistic analysis. H pylori infection was significantly associated with the MetS in HIV-infected patients.


Novel Analysis of Immune Cells from Nasal Microbiopsy Demonstrates Reliable, Reproducible Data for Immune Populations, and Superior Cytokine Detection Compared to Nasal Wash.
The morbidity and mortality related to respiratory tract diseases is enormous, with hundreds of millions of individuals afflicted and four million people dying each year. Understanding the immunological processes in the mucosa that govern outcome following pathogenic encounter could lead to novel therapies. There is a need to study responses at mucosal surfaces in humans for two reasons: (i) Immunological findings in mice, or other animals, often fail to translate to humans. (ii) Compartmentalization of the immune system dictates a need to study sites where pathogens reside. In this manuscript, we describe two novel non-invasive nasal mucosal microsampling techniques and their use for measuring immunological parameters: 1) using nasal curettes to collect cells from the inferior turbinate and; 2) absorptive matrices to collect nasal lining fluid. Both techniques were well tolerated and yielded reproducible and robust data. We demonstrated differences in immune populations and activation state in nasal mucosa compared to blood as well as compared to nasopharyngeal lumen in healthy adults. We also found superior cytokine detection with absorptive matrices compared to nasal wash. These techniques are promising new tools that will facilitate studies of the immunological signatures underlying susceptibility and resistance to respiratory infections.


The State of Emergency Medical Services (EMS) Systems in Africa.
Introduction Little is known about the existence, distribution, and characteristics of Emergency Medical Services (EMS) systems in Africa, or the corresponding epidemiology of prehospital illness and injury. A survey was conducted between 2013 and 2014 by distributing a detailed EMS system questionnaire to experts in paper and electronic versions. The questionnaire ascertained EMS systems' jurisdiction, operations, finance, clinical care, resources, and regulatory environment. The discovery of respondents with requisite expertise occurred in multiple phases, including snowball sampling, a review of published scientific literature, and a rigorous search of the Internet. The survey response rate was 46%, and data represented 49 of 54 (91%) African countries. Twenty-five EMS systems were identified and distributed among 16 countries (30% of African countries). There was no evidence of EMS systems in 33 (61%) countries. A total of 98,574,731 (8.7%) of the African population were serviced by at least one EMS system in 2012. The leading causes of EMS transport were (in order of decreasing frequency): injury, obstetric, respiratory, cardiovascular, and gastrointestinal complaints. Nineteen percent of African countries had government-financed EMS systems and 26% had a toll-free public access telephone number. Basic emergency medical technicians (EMTs) and Basic Life Support (BLS)-equipped ambulances were the most common cadre of provider and ambulance level, respectively (84% each). Emergency Medical Services systems exist in one-third of African countries. Injury and obstetric complaints are the leading African prehospital conditions. Only a minority (<9.0%) of Africans have coverage by an EMS system. Most systems were predominantly BLS, government operated, and fee-for-service. Mould-Millman NK , Dixon JM , Sefa N , Yancey A , Hollong BG , Hagahmed M , Ginde AA , Wallis LA . The state of Emergency Medical Services (EMS) systems in Africa. Prehosp Disaster Med. 2017;32(3):273-283.


The clinical effectiveness of sertraline in primary care and the role of depression severity and duration (PANDA): a pragmatic, double-blind, placebo-controlled randomised trial.
Depression is usually managed in primary care, but most antidepressant trials are of patients from secondary care mental health services, with eligibility criteria based on diagnosis and severity of depressive symptoms. Antidepressants are now used in a much wider group of people than in previous regulatory trials. We investigated the clinical effectiveness of sertraline in patients in primary care with depressive symptoms ranging from mild to severe and tested the role of severity and duration in treatment response. The PANDA study was a pragmatic, multicentre, double-blind, placebo-controlled randomised trial of patients from 179 primary care surgeries in four UK cities (Bristol, Liverpool, London, and York). We included patients aged 18 to 74 years who had depressive symptoms of any severity or duration in the past 2 years, where there was clinical uncertainty about the benefit of an antidepressant. This strategy was designed to improve the generalisability of our sample to current use of antidepressants within primary care. Patients were randomly assigned (1:1) with a remote computer-generated code to sertraline or placebo, and were stratified by severity, duration, and site with random block length. Patients received one capsule (sertraline 50 mg or placebo orally) daily for one week then two capsules daily for up to 11 weeks, consistent with evidence on optimal dosages for efficacy and acceptability. The primary outcome was depressive symptoms 6 weeks after randomisation, measured by Patient Health Questionnaire, 9-item version (PHQ-9) scores. Secondary outcomes at 2, 6 and 12 weeks were depressive symptoms and remission (PHQ-9 and Beck Depression Inventory-II), generalised anxiety symptoms (Generalised Anxiety Disorder Assessment 7-item version), mental and physical health-related quality of life (12-item Short-Form Health Survey), and self-reported improvement. All analyses compared groups as randomised (intention-to-treat). The study is registered with EudraCT, 2013-003440-22 (protocol number 13/0413; version 6.1) and ISRCTN, ISRCTN84544741, and is closed to new participants. Between Jan 1, 2015, and Aug 31, 2017, we recruited and randomly assigned 655 patients-326 (50%) to sertraline and 329 (50%) to placebo. Two patients in the sertraline group did not complete a substantial proportion of the baseline assessment and were excluded, leaving 653 patients in total. Due to attrition, primary outcome analyses were of 550 patients (266 in the sertraline group and 284 in the placebo group; 85% follow-up that did not differ by treatment allocation). We found no evidence that sertraline led to a clinically meaningful reduction in depressive symptoms at 6 weeks. The mean 6-week PHQ-9 score was 7·98 (SD 5·63) in the sertraline group and 8·76 (5·86) in the placebo group (adjusted proportional difference 0·95, 95% CI 0·85-1·07; p=0·41). However, for secondary outcomes, we found evidence that sertraline led to reduced anxiety symptoms, better mental (but not physical) health-related quality of life, and self-reported improvements in mental health. We observed weak evidence that depressive symptoms were reduced by sertraline at 12 weeks. We recorded seven adverse events-four for sertraline and three for placebo, and adverse events did not differ by treatment allocation. Three adverse events were classified as serious-two in the sertraline group and one in the placebo group. One serious adverse event in the sertraline group was classified as possibly related to study medication. Sertraline is unlikely to reduce depressive symptoms within 6 weeks in primary care but we observed improvements in anxiety, quality of life, and self-rated mental health, which are likely to be clinically important. Our findings support the prescription of SSRI antidepressants in a wider group of participants than previously thought, including those with mild to moderate symptoms who do not meet diagnostic criteria for depression or generalised anxiety disorder. National Institute for Health Research.


Acoustofluidic chemical waveform generator and switch.
Eliciting a cellular response to a changing chemical microenvironment is central to many biological processes including gene expression, cell migration, differentiation, apoptosis, and intercellular signaling. The nature and scope of the response is highly dependent upon the spatiotemporal characteristics of the stimulus. To date, studies that investigate this phenomenon have been limited to digital (or step) chemical stimulation with little control over the temporal counterparts. Here, we demonstrate an acoustofluidic (i.e., fusion of acoustics and microfluidics) approach for generating programmable chemical waveforms that permits continuous modulation of the signal characteristics including the amplitude (i.e., sample concentration), shape, frequency, and duty cycle, with frequencies reaching up to 30 Hz. Furthermore, we show fast switching between multiple distinct stimuli, wherein the waveform of each stimulus is independently controlled. Using our device, we characterized the frequency-dependent activation and internalization of the β2-adrenergic receptor (β2-AR), a prototypic G-protein coupled receptor (GPCR), using epinephrine. The acoustofluidic-based programmable chemical waveform generation and switching method presented herein is expected to be a powerful tool for the investigation and characterization of the kinetics and other dynamic properties of many biological and biochemical processes.


Perceived health status is associated with hours of exercise per week in older adults independent of physical health.
Perceived health status does not always reflect actual health status. We investigated the association between objective and self-rated measures of health status and hours of exercise per week in older adults. As part of the TRIL clinic assessment, we gathered information from 473 community dwelling adults over the age of 65, regarding hours spent per week exercising, depression, personality, perceived health status, and objective health status (in the form of a comorbidity count). Regression analyses were performed on these data to investigate whether perceived health status, objective health status, personality and mood are associated with hours of exercise per week. Perceived and objective health status were significantly but weakly correlated. Both perceived and objective health status, as well as depression, were independently associated with hours of exercise per week. We conclude that exercise uptake in older adults is contingent on both perceived and objective health status, as well as depression. Perceived health status has a stronger association with exercise uptake in older adults with lower depression levels. The current findings have implications for designing exercise interventions for older adults.


Structure of the basal components of a bacterial transporter.
Proteins SpoIIQ and SpoIIIAH interact through two membranes to connect the forespore and the mother cell during endospore development in the bacterium Bacillus subtilis. SpoIIIAH consists of a transmembrane segment and an extracellular domain with similarity to YscJ proteins. YscJ proteins form large multimeric rings that are the structural scaffolds for the assembly of type III secretion systems in gram-negative bacteria. The predicted ring-forming motif of SpoIIIAH and other evidence led to the model that SpoIIQ and SpoIIIAH form the core components of a channel or transporter through which the mother cell nurtures forespore development. Therefore, to understand the roles of SpoIIIAH and SpoIIQ in channel formation, it is critical to determine whether SpoIIIAH adopts a ring-forming structural motif, and whether interaction of SpoIIIAH with SpoIIQ would preclude ring formation. We report a 2.8-Å resolution structure of a complex of SpoIIQ and SpoIIIAH. SpoIIIAH folds into the ring-building structural motif, and modeling shows that the structure of the SpoIIQ-SpoIIIAH complex is compatible with forming a symmetrical oligomer that is similar to those in type III systems. The inner diameters of the two most likely ring models are large enough to accommodate several copies of other integral membrane proteins. SpoIIQ contains a LytM domain, which is found in metalloendopeptidases, but lacks residues important for metalloprotease activity. Other LytM domains appear to be involved in protein-protein interactions. We found that the LytM domain of SpoIIQ contains an accessory region that interacts with SpoIIIAH.


Bronchial gene expression signature associated with rate of subsequent FEV1 decline in individuals with and at risk of COPD.
COPD is characterised by progressive lung function decline. Leveraging prior work demonstrating bronchial airway COPD-associated gene expression alterations, we sought to determine if there are alterations associated with differences in the rate of FEV1 decline. We examined gene expression among ever smokers with and without COPD who at baseline had bronchial brushings profiled by Affymetrix microarrays and had longitudinal lung function measurements (n=134; mean follow-up=6.38±2.48 years). Gene expression profiles associated with the rate of FEV1 decline were identified by linear modelling. Expression differences in 171 genes were associated with rate of FEV1 decline (false discovery rate <0.05). The FEV1 decline signature was replicated in an independent dataset of bronchial biopsies from patients with COPD (n=46; p=0.018; mean follow-up=6.76±1.32 years). Genes elevated in individuals with more rapid FEV1 decline are significantly enriched among the genes altered by modulation of XBP1 in two independent datasets (Gene Set Enrichment Analysis (GSEA) p<0.05) and are enriched in mucin-related genes (GSEA p<0.05). We have identified and replicated an airway gene expression signature associated with the rate of FEV1 decline. Aspects of this signature are related to increased expression of XBP1-regulated genes, a transcription factor involved in the unfolded protein response, and genes related to mucin production. Collectively, these data suggest that molecular processes related to the rate of FEV1 decline can be detected in airway epithelium, identify a possible indicator of FEV1 decline and make it possible to detect, in an early phase, ever smokers with and without COPD most at risk of rapid FEV1 decline.


Validity of the Agency for Healthcare Research and Quality clinical practice guidelines: how quickly do guidelines become outdated?
Practice guidelines need to be up-to-date to be useful to clinicians. No published methods are available for assessing whether existing practice guidelines are still valid, nor does any empirical information exist regarding how often such assessments need to be made. To assess the current validity of 17 clinical practice guidelines published by the US Agency for Healthcare Research and Quality (AHRQ) that are still in circulation, and to use this information to estimate how quickly guidelines become obsolete. We developed criteria for defining when a guideline needs updating, mailed surveys to members of the original AHRQ guideline panels (n = 170; response rate, 71%), and searched the literature for evidence through March 2000 (n = 6994 titles yielding 173 articles plus 159 new guidelines on the same topics). Identification of new evidence calling for a major, minor, or no update of the 17 guidelines; survival analysis of the rate at which guidelines became outdated. For 7 guidelines, new evidence and expert judgment indicated that a major update is required; 6 were found to be in need of a minor update; 3 were judged as still valid; and for 1 guideline, we could reach no conclusion. Survival analysis indicated that about half the guidelines were outdated in 5.8 years (95% confidence interval [CI], 5.0-6.6 years). The point at which no more than 90% of the guidelines were still valid was 3.6 years (95% CI, 2.6-4.6 years). More than three quarters of the AHRQ guidelines need updating. As a general rule, guidelines should be reassessed for validity every 3 years.


Restoration of bowel continuity after surgery for acute perforated diverticulitis: should Hartmann's procedure be considered a one-stage procedure?
Hartmann's procedure (HP) still remains the most frequently performed procedure in acute perforated diverticulitis, but it results in a end colostomy. Primary anastomosis (PA) with or without defunctioning loop ileostomy (DI) seems a good alternative. The aim of this study was to assess differences in the rate of stomal reversal after HP and PA with DI and to evaluate factors associated with postreversal morbidity in patients operated for acute perforated diverticulitis. All 158 patients who had survived emergency surgery for acute perforated diverticulitis in five teaching hospitals in The Netherlands between 1995 and 2005 and underwent HP or PA with DI were retrospectively studied. Age, gender, ASA-classification, severity of primary disease, delay of stoma reversal, surgeon's experience, surgical procedure and type of anastomosis were analysed in relation to outcome after stoma reversal. Of the 158 patients, 139 had undergone HP and 19 PA with DI. The reversal-rate was higher in patients with DI (14/19; 74%) compared to HP (63/139; 45%) (P = 0.027) Delay between primary surgery and stoma reversal was shorter after PA with DI compared with HP (3.9 vs 9.1 months; P < 0.001). Cumulative postreversal morbidity after HP was 44%. Early surgical complications occurred in 22 of 63 patients. Morbidity after DI reversal was 15% (P < 0.001). Three patients died after HP reversal, none died after DI reversal. Anastomotic leakage was observed in 10 patients after HP reversal. This was less frequently observed when the operation was performed by a specialist colorectal surgeon (10%vs 33%; P = 0.049) and when a stapled anastomosis was performed (4%vs 24%; P = 0.037). Reversal of HP should only be performed by an experienced colorectal surgeon, preferably performing a stapled anastomosis, or probably not be performed at all, as it is accompanied by high postoperative morbidity and even mortality. It is important that these findings are taken in account for when performing primary emergency surgery for acute perforated diverticulitis.


Role of collectins and complement protein C1q in pregnancy and parturition.
Collectins such as surfactant proteins SP-A, SP-D, and mannan-binding lectin (MBL), as well as complement protein C1q are evolutionarily conserved innate immune molecules. They are known to opsonize a range of microbial pathogens (bacteria, fungi, virus, and parasites) and trigger effector clearance mechanisms involving phagocytosis and/or complement activation. Collectins and C1q have also attracted attention in studies involving pregnancy as they are expressed in the female reproductive tissues during pregnancy; a unique state of immune suppression with increased susceptibility to infectious diseases. Recent studies are beginning to unravel their functional significance in implantation, placentation, pregnancy maintenance and parturition in normal and adverse pregnancies. Collectins and C1q, expressed in gestational tissues during pregnancy, might alter the status of mother's immune response to the allogenic fetus and the microenvironment, thereby serving as important regulators of fetus-mother interaction. Here, we discuss the functional roles that have been assigned to SP-A, SP-D, MBL and C1q in pregnancy and parturition.


Influence of alcohol use experience and motivational drive on college students' alcohol-related cognition.
Cognitive processes are thought to be pivotal to risk for heavy drinking. However, few studies have examined the alcohol cue-activated positive and negative semantic memory networks that may be pivotal to drinking behavior. Moreover, much is to be understood about the influences of cognitive processes, particularly in high-risk drinking samples such as college students. The current study examines the sequential process of alcohol cues activating valenced semantic memory networks, and the influences of prior drinking experience and individual differences in motivational drive on this automatic (implicit) cognitive process. Participants (N = 138, 52% women) were college freshmen prescreened to represent the full range of drinking experience (i.e., current abstainers, light and heavy drinkers). Participants completed self-reports of past month alcohol use, and individual differences in behavioral inhibition system (BIS) and behavioral approach/activation system (BAS). Alcohol cue-elicited positive and negative semantic memory networks were assessed using a priming task. Results from the priming task revealed that for light drinkers alcohol cues were equally as likely to activate positive and negative semantic memory networks, suggesting relatively neutral cue-elicited alcohol attitudes. Conversely, for heavy drinkers, alcohol cues more readily activated positive relative to negative semantic memory networks, suggesting relatively positive cue-elicited alcohol attitudes. Furthermore, positive alcohol cue-elicited semantic memory networks (positive attitudes) were evident for individuals characterized by a strong BAS and weak BIS (as hypothesized) and those characterized by a weak BAS and weak BIS. The findings suggest that alcohol-cue elicited positive semantic memory networks may be pivotal to risk for heavy drinking. Specifically, it is via the influence on this cognitive process that prior drinking experience and individual differences in motivational drive, respectively, may maintain and predispose individuals to risk for heavy alcohol use.


Comorbidity and outcomes of concurrent chemo- and radiotherapy in limited disease small cell lung cancer.
Many patients with limited disease small cell lung cancer (LD SCLC) suffer from comorbidity. Not all patients with comorbidity are offered standard treatment, though there is little evidence for such a policy. The aim of this study was to investigate whether patients with comorbidity had inferior outcomes in a LD SCLC cohort. We analyzed patients from a randomized study comparing two three-week schedules of thoracic radiotherapy (TRT) plus standard chemotherapy in LD SCLC. Patients were to receive four courses of cisplatin/etoposide and TRT of 45 Gy/30 fractions (twice daily) or 42 Gy/15 fractions (once daily). Responders received prophylactic cranial irradiation (PCI). Comorbidity was assessed using the Charlson Comorbidity Index (CCI), which rates conditions with increased one-year mortality. In total 157 patients were enrolled between May 2005 and January 2011. Median age was 63 years, 52% were men, 16% had performance status 2, and 72% stage III disease. Forty percent had no comorbidity; 34% had CCI-score 1; 15% CCI 2; and 11% CCI 3-5. There were no significant differences in completion rates of chemotherapy, TRT or PCI across CCI-scores; or any significant differences in the frequency of grade 3-5 toxicity (p = 0.49), treatment-related deaths (p = 0.36), response rates (p = 0.20), progression-free survival (p = 0.18) or overall survival (p = 0.09) between the CCI categories. Patients with comorbidity completed and tolerated chemo-radiotherapy as well as other patients. There were no significant differences in response rates, progression-free survival or overall survival - suggesting that comorbidity alone is not a reason to withhold standard therapy in LD SCLC.


International external quality assessment study for molecular detection of Lassa virus.
Lassa virus (LASV) is a causative agent of hemorrhagic fever in West Africa. In recent years, it has been imported several times to Europe and North America. The method of choice for early detection of LASV in blood is RT-PCR. Therefore, the European Network for Diagnostics of 'Imported' Viral Diseases (ENIVD) performed an external quality assessment (EQA) study for molecular detection of LASV. A proficiency panel of 13 samples containing various concentrations of inactivated LASV strains Josiah, Lib-1580/121, CSF, or AV was prepared. Samples containing the LASV-related lymphocytic choriomeningitis virus (LCMV) and negative sera were included as specificity controls. Twenty-four laboratories from 17 countries (13 European, one African, one Asian, two American countries) participated in the study. Thirteen laboratories (54%) reported correct results, 4 (17%) laboratories reported 1 to 2 false-negative results, and 7 (29%) laboratories reported 3 to 5 false-negative results. This EQA study indicates that most participating laboratories have a good or acceptable performance in molecular detection of LASV. However, several laboratories need to review and improve their diagnostic procedures.


The Bergen Shopping Addiction Scale: reliability and validity of a brief screening test.
Although excessive and compulsive shopping has been increasingly placed within the behavioral addiction paradigm in recent years, items in existing screens arguably do not assess the core criteria and components of addiction. To date, assessment screens for shopping disorders have primarily been rooted within the impulse-control or obsessive-compulsive disorder paradigms. Furthermore, existing screens use the terms 'shopping,' 'buying,' and 'spending' interchangeably, and do not necessarily reflect contemporary shopping habits. Consequently, a new screening tool for assessing shopping addiction was developed. Initially, 28 items, four for each of seven addiction criteria (salience, mood modification, conflict, tolerance, withdrawal, relapse, and problems), were constructed. These items and validated scales (i.e., Compulsive Buying Measurement Scale, Mini-International Personality Item Pool, Hospital Anxiety and Depression Scale, Rosenberg Self-Esteem Scale) were then administered to 23,537 participants (M age = 35.8 years, SD age = 13.3). The highest loading item from each set of four pooled items reflecting the seven addiction criteria were retained in the final scale, The Bergen Shopping Addiction Scale (BSAS). The factor structure of the BSAS was good (RMSEA = 0.064, CFI = 0.983, TLI = 0.973) and coefficient alpha was 0.87. The scores on the BSAS converged with scores on the Compulsive Buying Measurement Scale (CBMS; 0.80), and were positively correlated with extroversion and neuroticism, and negatively with conscientiousness, agreeableness, and intellect/imagination. The scores of the BSAS were positively associated with anxiety, depression, and low self-esteem and inversely related to age. Females scored higher than males on the BSAS. The BSAS is the first scale to fully embed shopping addiction within an addiction paradigm. A recommended cutoff score for the new scale and future research directions are discussed.


Dynamic assembly of ribbon synapses and circuit maintenance in a vertebrate sensory system.
Ribbon synapses transmit information in sensory systems, but their development is not well understood. To test the hypothesis that ribbon assembly stabilizes nascent synapses, we performed simultaneous time-lapse imaging of fluorescently-tagged ribbons in retinal cone bipolar cells (BCs) and postsynaptic densities (PSD95-FP) of retinal ganglion cells (RGCs). Ribbons and PSD95-FP clusters were more stable when these components colocalized at synapses. However, synapse density on ON-alpha RGCs was unchanged in mice lacking ribbons (ribeye knockout). Wildtype BCs make both ribbon-containing and ribbon-free synapses with these GCs even at maturity. Ribbon assembly and cone BC-RGC synapse maintenance are thus regulated independently. Despite the absence of synaptic ribbons, RGCs continued to respond robustly to light stimuli, although quantitative examination of the responses revealed reduced frequency and contrast sensitivity.


Pattern of breathing during progressive exercise in chronic heart failure.
This descriptive study analyzed serial, individual changes in the exercise pattern of breathing (POB) of patients with stable chronic heart failure (CHF). Twenty-two CHF patients underwent maximal, symptom-limited cardiopulmonary exercise test on a treadmill. Minute ventilation (VE), tidal volume (VT), breathing frequency (f), the ventilatory equivalent for carbon dioxide (VE/VCO2) and estimated dead-space to tidal volume ratio (VD/VT) were continuously recorded. The VE/VCO2 slope was calculated in every subject as the slope of the regression line relating VE to VCO2 during exercising testing. Pattern of breathing was investigated by constructing the individual VT-f relationship for each patient separately. In 16 (73%) patients (group 1), the VT-f plot was initially linear, but subsequently exhibited an inflection point at which VT stopped increasing with further increases in f. In six (27%) patients (group 2) no inflection point was evident on the VT-f relation; in four of these patients the VT-f relation remained linear but shifted to the right throughout testing, and two patients decreased VT before peak exercise achieving high breathing frequencies. Comparing group 1 to group 2 patients, they had higher VEmax (68+/-23 vs. 44+/-10 l/min, P=0.02) and VO2max (17+/-5 vs. 12+/-3 ml/min/kg, P=0.01). In contrast, the two groups did not differ in terms of age, weight, height, diagnosis, ejection fraction or VE/VCO2 slope. In conclusion, patients with CHF adopt variable breathing patterns during exercise; specific patterns are associated with greater impairment in functional capacity.


Differential response of MC3T3-E1 and human mesenchymal stem cells to inositol hexakisphosphate.
Inositol hexakisphosphate (IP6) has been found to have an important role in biomineralization. Because the complete mechanism of action of IP6 on osteoblasts is not fully understood and its potential use in the primary prevention of osteoporosis, we examined the direct effect of IP6 on cell viability and differentiation of MC3T3-E1 cells and on differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs). We show that IP6 has different effects depending on the origin of the cell target. Thus, while IP6 decreased gene expression of osteoblast markers and mineralization in MC3T3-E1 cells without negatively affecting cell viability and ALP activity, an increase in gene expression of ALP was observed in hUC-MSCs committed to the osteoblastic lineage. This increasing effect of IP6 on ALP mRNA expression levels was reversed by the addition of a selective inhibitor of IP6 kinase, suggesting that the effect of IP6 might be due through its pyrophosphorylated derivatives. Besides, Rankl mRNA levels were decreased after IP6 treatment in MC3T3-E1 cells, pointing to a paracrine effect on osteoclasts. Our results indicate that IP6 has different effects on osteoblast differentiation depending on the cell type and origin. However, further studies are needed to examine the net effect of IP6 on bone formation and its potential as novel antiosteoporosis drug.


Elevated IgG autoantibody production in oligoarticular juvenile idiopathic arthritis may predict a refractory course.
Although oligoarticular juvenile idiopathic arthritis (oJIA) is considered to carry the best prognosis among the JIA subtypes, many children evolve to a chronic course. A few studies have identified clinical risk factors for disease extension, and recent studies have evaluated synovial fluid markers. However, the only biological marker from the serum studied to date is the anti-nuclear antibody (ANA), regarding which there is mixed data regarding prognosis. No studies have evaluated whether additional autoantibodies may affect the articular prognosis of oJIA. Microarrays containing candidate autoantigens were printed on slides, which were used to profile 36 children with oJIA and 18 controls. Unsupervised cluster analysis was used to identify distinct subgroups of JIA patients. Response to therapy after a mean interval of 4.9 months was evaluated. Cluster analysis revealed two subgroups of oJIA patients, with identical clustering observed when children with onset over age six were excluded. Cluster 1 had higher levels of multiple autoantibodies compared to both cluster 2 as well as controls, including antibodies against several extracellular matrix (ECM) and nuclear antigens. Although the two patient clusters were similar with respect to clinical features and treatment decisions, children in cluster 1 were less likely to have attained remission by the follow-up visit. Antibodies against ECM and possibly other antigens may identify a sub-group of children with oJIA who will require more aggressive therapy to attain control of the arthritis.


Determinants of Visceral Leishmaniasis: A Case-Control Study in Gedaref State, Sudan.
Improving knowledge on local determinants of visceral leishmaniasis (VL) is crucial to guide the development of relevant control strategies. This study aimed to identify individual and household level determinants of primary VL in 24 highly endemic villages of Tabarak Allah hospital's catchment area, Gedaref State, Sudan. From September 2012 to July 2013, in an unmatched case-control design, 198 patients with primary VL were compared to 801 controls free of VL symptoms and with a negative VL rapid test. Using random spatial sampling, controls were selected with a distribution of age, sex and village of residence proportionate to the distribution of the target population. Data were collected using a structured questionnaire. Children and men were at higher risk of VL. Reporting VL patient(s) in the household in the previous year was the strongest VL risk factor. In a multivariate analysis, VL risk increased with household size, sleep location (outside the yard, not in the farm), evening outdoor activities in the rainy season (playing, watching TV, radio listening), use of ground nut oil as animal repellent and of smoke of Acacia seyal as indoor repellent, presence of dogs in the yard at night, Acacia nilotica in the yard's immediate surroundings and of a forest at eye range. VL risk appeared to decrease with the use of drinking water sources other than the village water tank, a buffer distance from the adjacent house yard, and with the presence of animals other than dogs in the yard at night. In contrast with previous studies, housing factors, mosquito-net use, black cotton soil, ethnicity, socioeconomic index, presence of Balanites aegyptica and Azadirachta indica in the yard were not independent VL determinants. Although these results do not provide evidence of causality, they provide useful suggestions for guiding further intervention studies on VL preventive measures.


Low Temperature Fluorescence Imaging of Freeze-trapped Human Cervical Tissues.
We characterized the fluorescence intensity distribution within the epithelia and stroma of frozen human cervical tissues at the following excitation-emission wavelength pairs: 440, 525 nm and 365, 460 nm. The intensities at both excitation-emission wavelength pairs are significantly lower in the epithelia of severely dysplastic tissues, relative to that in normal and inflammatory tissues. Furthermore, there are small differences in (1) the epithelial intensity of severe dysplasia and mild dysplasia at 440, 525 nm and (2) the stromal intensity of inflammatory and severely dysplastic tissues at 365, 460 nm. A comparison of the ratio of intensities at 440, 525 nm and 365, 460 nm between the epithelia of each tissue type indicates that this ratio is lowest in severely dysplastic tissues. It is interesting to note that the epithelial and stromal intensities are comparable at 365, 460 nm; however, at 440, 525 nm, the epithelial intensity is more than a factor of two less that that of the stroma for all tissue types.


Can genetic polymorphisms predict response variability to anodal transcranial direct current stimulation of the primary motor cortex?
Genetic mediation of cortical plasticity and the role genetic variants play in previously observed response variability to transcranial direct current stimulation (tDCS) have become important issues in the tDCS literature in recent years. This study investigated whether inter-individual variability to tDCS was in-part genetically mediated. In 61 healthy males, anodal-tDCS (a-tDCS) and sham-tDCS were administered to the primary motor cortex at 1 mA for 10-min via 6 × 4 cm active and 7 × 5 cm return electrodes. Twenty-five single-pulse transcranial magnetic stimulation (TMS) motor evoked potentials (MEP) were recorded to represent corticospinal excitability (CSE). Twenty-five paired-pulse MEPs were recorded with 3 ms inter-stimulus interval (ISI) to assess intracortical inhibition (ICI) via short-interval intracranial inhibition (SICI) and 10 ms ISI for intracortical facilitation (ICF). Saliva samples were tested for specific genetic polymorphisms in genes encoding for excitatory and inhibitory neuroreceptors. Individuals were sub-grouped based on a pre-determined threshold and via statistical cluster analysis. Two distinct subgroups were identified, increases in CSE following a-tDCS (i.e. Responders) and no increase or even reductions in CSE (i.e. Non-responders). No changes in ICI or ICF were reported. No relationships were reported between genetic polymorphisms in excitatory receptor genes and a-tDCS responders. An association was reported between a-tDCS responders and GABRA3 gene polymorphisms encoding for GABA-A receptors suggesting potential relationships between GABA-A receptor variations and capacity to undergo tDCS-induced cortical plasticity. In the largest tDCS study of its kind, this study presents an important step forward in determining the contribution genetic factors play in previously observed inter-individual variability to tDCS.