Results of intrastromal corneal ring implantation in advanced keratoconus With the aim of postpoing or avoiding corneal transplantation.
To evaluate the safety and efficacy of the surgery of intracorneal ring segment implantation with 320° of arc (320-ICRS) in patients with advanced keratoconus stage IV and maximum keratometry (Kmax) above 60 D. A prospective, interventional case series study evaluating 25 eyes of 19 patients with keratoconus stage IV and Kmax > 60D in which 320-ICRS were implanted using VisuMax® femtosecond. Uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), keratometric values (mean - mean-K, flat - K1, and steep - K2), maximum keratometry (Kmax), tomographic astigmatism, refractive astigmatism and asphericity (Q) were assessed preoperatively and at 3, 6 and 12 months after the procedure. The UDVA improved from 1.03 ± 0.28 LogMAR (20/200) to 0.54 ± 0.21 LogMAR (20/60), (p < 0.001), the CDVA (with glasses) improved from 0.63 ± 0.29 LogMAR (20/80) to 0.31 ± 0.16 LogMAR (20/40),(p = 0.004), K1 reduced from 54.41 ± 4.46 D to 49.36 ± 4.11 D (p < 0.001), K2 reduced from 61.15 ± 4.37 D to 53.715 ± 4.05 D, (p < 0.001), mean-K reduced from 57.55 ± 4,17 D to 51.44 ± 3,94 D (p < 0.001), Kmax reduced from 69.80 ± 8.20 D to 63.43 ± 6.31 D (p < 0.001) and asphericity (Q) changed from -1.57 ± 0.35 to -0.77 ± 0.56 (p < 0.001). A total of 89.9% patients reached BCVA wearing scleral contact lens 0.2 LogMAR(20/25). 320-ICRS to treat advanced keratoconus appears to be an efficacious and safe procedure, being a surgical alternative to delay or even prevent corneal transplantation.


Posture interacts with arm weight support to modulate corticomotor excitability to the upper limb.
The use of arm weight support (WS) to optimize movement quality may be an avenue for improved upper limb stroke rehabilitation; however, the underlying neurophysiological effects of WS are not well understood. Rehabilitation exercises may be performed when sitting or standing, but the interaction of posture with WS has not been examined until now. We explored the effect of posture with WS on corticomotor excitability (CME) in healthy adults. Thirteen participants performed static shoulder abduction in two postures (sitting and standing) at three levels of WS (0, 45, and 90 % of full support). Transcranial magnetic stimulation of primary motor cortex was used to elicit motor-evoked potentials (MEPs) in eight upper limb muscles. Stimulus-response (SR) curves were fitted to the MEP data using nonlinear regression. Whole-body posture interacted with WS to influence tonic activity and CME in all muscles examined. SR curve parameters revealed greater CME when standing compared to sitting for upper arm muscles, but lower CME to the shoulder, forearm, and hand. Distal to the shoulder, tonic activity and CME were modulated independent of any explicit differences in task requirements. Overall, these results support a model of integrated upper limb control influenced by whole-body posture and WS. These findings have implications for the application of WS in settings such as upper limb rehabilitation after stroke.


Short-term radiorespirometry of cell suspensions.
1. An apparatus and method are described with which the oxidation of labelled substrates to (14)CO(2) by cell suspensions may be examined. 2. The use of high-specific-radioactivity substrates at low concentration together with frequent quantitative collection of CO(2) permit a more detailed analysis of the appearance of various substrate carbon atoms as CO(2) than is possible by existing techniques. 3. Typical experiments with various cell types are reported, in which pathways of glucose oxidation are examined.


Effect of flubendazole on Cysticercus cellulosae in pigs.
The effect of flubendazole on Cysticercus cellulosae was tested in 15 pigs. In doses o 8.3 mg/kg body weight or more daily for 10 days the drug was lethal to the cysticerci, causing both macroscopic and microscopic morphological alterations. An in vitro test showed 0% viability of cysticerci in most of the treated pigs, whereas in 90-100% of 11 control pigs they were viable.


Sexual, ontogenetic, temporal and spatial effects on the diet of Urotrygon rogersi (Elasmobranchii:Myliobatiformes).
The food habits and trophic ecology of Urotrygon rogersi were analysed to ascertain sexual, ontogenetic, temporal and spatial effects on the diet. These effects were examined with contingency tables, simple correspondence analysis and MANOVA. The trophic relationships of the species were determined with Levin's niche amplitude index and Pianka's diet overlap index. Urotrygon rogersi is a predator of benthic organisms, especially shrimp and polychaetes. Analysis of sex and time showed no significant differences, but differences among class sizes were found. A strong trend towards diet specialization and a strong overlap between sexes and between class sizes were found. Finally, fishing activity on shrimps in the study area could force U. rogersi to change its trophic spectrum over time as an adaptation to the considerable reduction in its principal prey.


Proteomic Characterization of the Heart and Skeletal Muscle Reveals Widespread Arginine ADP-Ribosylation by the ARTC1 Ectoenzyme.
The clostridium-like ecto-ADP-ribosyltransferase ARTC1 is expressed in a highly restricted manner in skeletal muscle and heart tissue. Although ARTC1 is well studied, the identification of ARTC1 targets in vivo and subsequent characterization of ARTC1-regulated cellular processes on the proteome level have been challenging and only a few ARTC1-ADP-ribosylated targets are known. Applying our recently developed mass spectrometry-based workflow to C2C12 myotubes and to skeletal muscle and heart tissues from wild-type mice, we identify hundreds of ARTC1-ADP-ribosylated proteins whose modifications are absent in the ADP-ribosylome of ARTC1-deficient mice. These proteins are ADP-ribosylated on arginine residues and mainly located on the cell surface or in the extracellular space. They are associated with signal transduction, transmembrane transport, and muscle function. Validation of hemopexin (HPX) as a ARTC1-target protein confirmed the functional importance of ARTC1-mediated extracellular arginine ADP-ribosylation at the systems level.


[Dependence of the intensity of growth processes on the potential difference at the cytoplasmic membrane].
The growth of organisms is a dynamic process of interrelated fluxes of substances and energy which most often proceeds in the steady-state conditions, the energy-dependent process being predominant. The rate of growth is directly proportional to the potential difference at the plasma membrane, created essentially by a passive influx of energy substance to the cell oxidative system which is a limiting link of its energization. The formation of potential difference is influenced by concentrations of biomass and energy substrate at the plasma membrane, as well as the values of electric capacity of the membrane and rate constants for the electric charge formation and expenditure.


Distribution of a specific 500-base-pair fragment in mycobacterium bovis isolates from Sardinian cattle.
Amplification of a specific, 500-bp fragment from Mycobacterium bovis isolates and use of the fragment to differentiate between Mycobacterium tuberculosis and M. bovis was previously reported (J. G. Rodriguez, G. A. Meja, P. Del Portillo, M. E. Patarroyo, and L. A. Murillo, Microbiology 141:2131-2138, 1995). In the present study, 30 M. bovis isolates from Sardinian cattle were examined for the presence of this 500-bp fragment; 4 of the 30 isolates lacked the fragment. This result indicates that identification of M. bovis strains by amplification of the 500-bp sequence may lead to false-negative results.


Designing a potent L1 protein-based HPV peptide vaccine: A bioinformatics approach.
Oncogenic human papilloma viruses (HPV) are the cause of various types of cancer, specifically cervical cancer. L1 protein is the main protein of HPV capsid which targeted in many vaccine-producing attempts. However, they have not enough coverage on the various high risk HPV types. Therefore, having a low cost potent HPV vaccine to protect against all members of the α-papillomaviridea family will be promising. In this study, L1 protein-based peptide vaccine was designed using immunoinformatics methods which provides physicochemical properties such as stability in room temperature, potential of antigenicity, non-allergic properties and no requirement with eukaryotic host system. The designed vaccine has two HPV conserved epitopes with lengths 18 and 27 amino acids in all members of α-papillomaviridea. These peptides promote humoral and cellular immunity and INF-γ responses. In order to ensure strong induction of immune responses, Flagellin, a Toll like receptor 5(TLR-5) agonist, and a short synthetic toll like receptor 4 (TLR-4) agonist were also joined to the epitopes. Structure of the designed- vaccine was validated using Rampage and ERRAT and a high quality 3D structure of the vaccine protein was provided. Docking studies demonstrated an appropriate and stable interaction between the vaccine and TLR-5. The vaccine is expected to have a high quality structure and suitable properties including high stability, solubility and a high potential to be expressed in E.coli. High potentiality of the vaccine in inducing humoral and cellular immune responses, may be considered as an anti-tumor vaccine.


[Radiotherapy: metrological provision].
The accuracy of delivering the dose of radiation during the therapy is determined by many factors. Its enhancement requires substantial material costs. All the requirements for enhancing the accuracy are shown to be outrageous as the proportion of negative results of treatment virtually does not vary with the dosimetric and technical equipping of radiation therapy. This must alter its technical equipping policy without increasing the requirements for the accuracy of dosimetric measurements.


Antimicrobial efficacy of orthodontic primer added with silver nanoparticles. Cross-sectional in vitro study.
The antimicrobial efficacy of the silver nanoparticles (NPsAg), incorporated into the adhesive (primer) placed in the enamel adjacent to fixed orthodontic appliances (brackets), was evaluated. An experimental study was performed on 40 premolars in vitro, which were divided into two groups with brackets, one cemented with conventional primer and another added with NPsAg, placed in culture media previously inoculated with Streptococcus mutans, and sampled for culturing and counting colony forming units (UFC) on days 1, 15 and 30. A decrease in the presence of Streptococcus mutans in the samples after 15 days with nanoparticle aggregation was observed, and a reduction in the effect of said nanoparticles after 30 days. This reduction of the nanoparticles effects can be due to the absence of mechanical cleaning, which favored the bacterial aggregation on the biofilm, affecting its antimicrobial effect. This suggest the need for realizing studies in "vivo" which will allow the observation of the behavior of the biometals on the buccal medium. The NPsAg added to the primer are an effective tool to prevent the demineralization of the enamel around the fixed orthodontic appliances.


Estimating individual health-related quality of life changes in low back pain patients.
There is a need to evaluate different options for estimating individual change in health-related quality of life for patients with low back pain. Secondary analysis of data collected at baseline and 6 weeks later in a randomized trial of 749 adults with low back pain receiving usual medical care (UMC) or UMC plus chiropractic care at a small hospital at a military training site or two large military medical centers. The mean age was 31; 76% were male and 67% were White. The study participants completed the Patient-Reported Outcomes Measurement Information System (PROMIS®)-29 v 1.0 physical function, pain interference, pain intensity, fatigue, sleep disturbance, depression, anxiety, satisfaction with participation in social roles, physical summary, and mental health summary scores (T-scored with mean = 50 and standard deviation (SD) = 10 in the U.S. general population). Reliability estimates at the baseline ranged from 0.700 to 0.969. Six-week test-retest intraclass correlation estimates were substantially lower than these estimates: the median test-retest intraclass correlation for the two-way mixed-effects model was 0. 532. Restricting the test-retest reliability estimates to the subset who reported they were about the same as at baseline on a retrospective rating of change item increased the median test-retest reliability to 0.686. The amount of individual change that was statistically significant varied by how reliability was estimated, and which SD was used. The smallest change needed was found when internal consistency reliability and the SD at baseline were used. When these values were used, the amount of change needed to be statistically significant (p < .05) at the individual level ranged from 3.33 (mental health summary scale) to 12.30 (pain intensity item) T-score points. We recommend that in research studies estimates of the magnitude of individual change needed for statistical significance be provided for multiple reliability and standard deviation estimates. Whenever possible, patients should be classified based on whether they 1) improved significantly and perceived they got better, 2) improved significantly but did not perceive they were better, 3) did not improve significantly but felt they got better, or 4) did not improve significantly or report getting better.


GaN nanowire tips for nanoscale atomic force microscopy.
Imaging of high-aspect-ratio nanostructures with sharp edges and straight walls in nanoscale metrology by atomic force microscopy (AFM) has been challenging due to the mechanical properties and conical geometry of the majority of available commercial tips. Here we report on the fabrication of GaN probes for nanoscale metrology of high-aspect-ratio structures to enhance the resolution of AFM imaging and improve the durability of AFM tips. GaN nanowires were fabricated using bottom-up and top-down techniques and bonded to Si cantilevers to scan vertical trenches on Si substrates. Over several scans, the GaN probes demonstrated excellent durability while scanning uneven structures and showed resolution enhancements in topography images, independent of scan direction, compared to commercial Si tips.


Controlling Charge Carrier Dynamics in Porphyrin Nanorings by Optically Active Templates.
Understanding the dynamics of photogenerated charge carriers is essential for enhancing the performance of solar and optoelectronic devices. Using atomistic quantum dynamics simulations, we demonstrate that a short π-conjugated optically active template can be used to control hot carrier relaxation, charge carrier separation, and carrier recombination in light-harvesting porphyrin nanorings. Relaxation of hot holes is slowed by 60% with an optically active template compared to that with an analogous optically inactive template. Both systems exhibit subpicosecond electron transfer from the photoactive core to the templates. Notably, charge recombination is suppressed 6-fold by the optically active template. The atomistic time-domain simulations rationalize these effects by the extent of electron and hole localization, modification of the density of states, participation of distinct vibrational motions, and changes in quantum coherence. Extension of the hot carrier lifetime and reduction of charge carrier recombination, without hampering charge separation, demonstrate a strategy for enhancing efficiencies of energy materials with optically active templates.


QTL mapping of adult-plant resistances to stripe rust and leaf rust in Chinese wheat cultivar Bainong 64.
Stripe rust and leaf rust, caused by Puccinia striiformis Westend. f. sp. tritici Erikss. and P. triticina, respectively, are devastating fungal diseases of common wheat (Triticum aestivum L.). Chinese wheat cultivar Bainong 64 has maintained acceptable adult-plant resistance (APR) to stripe rust, leaf rust and powdery mildew for more than 10 years. The aim of this study was to identify quantitative trait loci/locus (QTL) for resistance to the two rusts in a population of 179 doubled haploid (DH) lines derived from Bainong 64 × Jingshuang 16. The DH lines were planted in randomized complete blocks with three replicates at four locations. Stripe rust tests were conducted using a mixture of currently prevalent P. striiformis races, and leaf rust tests were performed with P. triticina race THTT. Leaf rust severities were scored two or three times, whereas maximum disease severities (MDS) were recorded for stripe rust. Using bulked segregant analysis (BSA) and simple sequence repeat (SSR) markers, five independent loci for APR to two rusts were detected. The QTL on chromosomes 1BL and 6BS contributed by Bainong 64 conferred resistance to both diseases. The loci identified on chromosomes 7AS and 4DL had minor effects on stripe rust response, whereas another locus, close to the centromere on chromosome 6BS, had a significant effect only on leaf rust response. The loci located on chromosomes 1BL and 4DL also had significant effects on powdery mildew response. These were located at the same positions as the Yr29/Lr46 and Yr46/Lr67 genes, respectively. The multiple disease resistance locus for APR on chromosome 6BS appears to be new. All three genes and their closely linked molecular markers could be used in breeding wheat cultivars with durable resistance to multiple diseases.


Log-Linear Modeling of Agreement among Expert Exposure Assessors.
Evaluation of expert assessment of exposure depends, in the absence of a validation measurement, upon measures of agreement among the expert raters. Agreement is typically measured using Cohen's Kappa statistic, however, there are some well-known limitations to this approach. We demonstrate an alternate method that uses log-linear models designed to model agreement. These models contain parameters that distinguish between exact agreement (diagonals of agreement matrix) and non-exact associations (off-diagonals). In addition, they can incorporate covariates to examine whether agreement differs across strata. We applied these models to evaluate agreement among expert ratings of exposure to sensitizers (none, likely, high) in a study of occupational asthma. Traditional analyses using weighted kappa suggested potential differences in agreement by blue/white collar jobs and office/non-office jobs, but not case/control status. However, the evaluation of the covariates and their interaction terms in log-linear models found no differences in agreement with these covariates and provided evidence that the differences observed using kappa were the result of marginal differences in the distribution of ratings rather than differences in agreement. Differences in agreement were predicted across the exposure scale, with the likely moderately exposed category more difficult for the experts to differentiate from the highly exposed category than from the unexposed category. The log-linear models provided valuable information about patterns of agreement and the structure of the data that were not revealed in analyses using kappa. The models' lack of dependence on marginal distributions and the ease of evaluating covariates allow reliable detection of observational bias in exposure data.


Comparative genomic analysis of coffee-infecting Xylella fastidiosa strains isolated from Brazil.
Strains of Xylella fastidiosa constitute a complex group of bacteria that develop within the xylem of many plant hosts, causing diseases of significant economic importance, such as Pierce's disease in North American grapevines and citrus variegated chlorosis in Brazil. X. fastidiosa has also been obtained from other host plants, in direct correlation with the development of diseases, as in the case of coffee leaf scorch (CLS)--a disease with potential to cause severe economic losses to the Brazilian coffee industry. This paper describes a thorough genomic characterization of coffee-infecting X. fastidiosa strains, initially performed through a microarray-based approach, which demonstrated that CLS strains could be subdivided in two phylogenetically distinct subgroups. Whole-genomic sequencing of two of these bacteria (one from each subgroup) allowed identification of ORFs and horizontally transferred elements (HTEs) that were specific to CLS-related X. fastidiosa strains. Such analyses confirmed the size and importance of HTEs as major mediators of chromosomal evolution amongst these bacteria, and allowed identification of differences in gene content, after comparisons were made with previously sequenced X. fastidiosa strains, isolated from alternative hosts. Although direct experimentation still needs to be performed to elucidate the biological consequences associated with such differences, it was interesting to verify that CLS-related bacteria display variations in genes that produce toxins, as well as surface-related factors (such as fimbrial adhesins and LPS) that have been shown to be involved with recognition of specific host factors in different pathogenic bacteria.


siRNA liposome-gold nanorod vectors for multispectral optoacoustic tomography theranostics.
Therapeutic applications of gene silencing using siRNA have seen increasing interest over the past decade. The optimization of the delivery and biodistribution of siRNA using liposome-gold nanorod (AuNRs) nanoscale carriers can greatly benefit from adept imaging methods that can visualize the time-resolved delivery performance of such vectors. In this work, we describe the effect of AuNR length incorporated with liposomes and show their complexation with siRNA as a novel gene delivery vehicle. We demonstrate the application of multispectral optoacoustic tomography (MSOT) to longitudinally visualize the localisation of siRNA carrying liposome-AuNR hybrids within tumors. Combination of in vivo MSOT with ex vivo fluorescence cryo-slice imaging offers further insight into the siRNA transport and activity obtained.


Effects of Mobility and Cognition on Risk of Mortality in Women in Late Life: A Prospective Study.
This study examines the effects of mobility and cognition on mortality risk in women late in life. A prospective study was conducted among 1,495 women (mean age 87.6 years) participating in the Study of Osteoporotic Fractures Year 20 examination (2006-2008). Mobility (ascertained by Short Physical Performance Battery [SPPB]) was categorized as poor (SPPB 0-3, n = 312), intermediate (SPPB 4-9, n = 799), or good (SPPB 10-12, n = 384). Cognitive status (adjudicated based on neuropsychological tests) was classified as normal (n = 873), mild cognitive impairment (n = 354), or dementia (n = 268). Deaths (n = 749) were identified from Year 20 through July 31, 2014 (average follow-up 4.9 years). There was not strong evidence of an interaction between mobility and cognition for prediction of mortality risk (p interaction term .16). Compared to women with good mobility, mortality risks were increased among women with intermediate mobility (hazard ratio [HR] 1.26, 95% confidence interval [CI] 1.02-1.57) and those with poor mobility (HR 1.64, 95% CI 1.24-2.16) after consideration of cognition and other mortality risk factors. Similarly, mortality risks were higher among women with mild cognitive impairment (HR 1.46, 95% CI 1.21-1.76) and those with dementia (HR 1.88, 95% CI 1.54-2.31) compared to women with normal cognition after consideration of mobility and other mortality risk factors. Among women late in life, 5-year mortality risk was substantially increased among women with deficits in mobility even after accounting for cognition and traditional prognostic indicators. Similarly, deficits in cognition were associated with increased 5-year mortality despite consideration of mobility and conventional risk factors.


Regulation of Embryonic Signal on Talin1 in Mouse Endometrium.
Embryonic signals can affect the spatiotemporal-specific expression of the uterus to establish a successful pregnancy. Our previous study has found that talin1 underwent dynamic changes in the mouse endometrium during peri-implantation period. However, whether talin1 is affected by the embryo signals is not clear. In order to investigate the effect of embryonic signals, especially human chorionic gonadotropin (HCG) on talin1, we have designed mouse models of pseudopregnancy, delayed implantation and activation, and HCG treatment. Using these models, the expression of talin1 in the mouse endometrium was determined by immunohistochemistry and Western blotting. In the pseudopregnancy model, an increased expression of talin1 was found from day 3 to day 5, whereas the talin1 protein was decreased on day 5 in the normal pregnant mice. In the delayed implantation model, a strong cytoplasmic staining of talin1 was found, especially in stromal cells. However, after activation of the implantation, the expression of talin1 decreased (P < .05). Furthermore, a significantly lower expression of talin1 was found at the implantation site when compared to the interimplantation sites (P < .05). In the HCG treatment model, an intrauterine perfusion of 10u HCG significantly reduced the expression of talin1 in both stromal and epithelial cells in pseudopregnant mice (P < .05), although further increase in the HCG concentration did not have additional effect on expression of talin1. Taken together, our data suggest that the presence of embryos can affect expression of talin1 in the mouse endometrium, and a certain concentration of HCG can regulate its expression.


Review of electronic cigarettes as tobacco cigarette substitutes: Their potential human health impact.
Electronic cigarettes (ECs) are devised to deliver nicotine in a vapor rather than in smoke without tar. ECs are hence advertised as being safer than tobacco cigarette products as the chemical compounds inhaled in the former are believed to be fewer and less toxic than those of the latter. Hazardous chemicals (e.g., formaldehyde) are nonetheless found to be generated incidentally by contacting the heated wire (i.e., the oxidation of glycerol/glycol in e-liquid). Although the extent of their release varies by several variables (e.g., the type of e-liquid, puffing rate, and the battery voltage), their exposure may also contribute to negative health effects. As the use of ECs may be much safer than that of common tobacco products, the former can be used as an aid to cut down or quit the latter. However, relatively little is yet known about the health effects of the EC on a long-term basis. Moreover, the use of EC cannot be clearly substantiated for renormalizing smoking behavior by current evidence. Behavior studies of the EC consumer suggest that the sufficient data for aerosol generation and chemical analysis should be acquired to establish reliable guides for its composition and consumption. In light of the urgent demand for such guidelines, this review examines the basic aspects of EC-related pollutants and their health effects.


Impact of myelin-specific antigen presenting B cells on T cell activation in multiple sclerosis.
The role of B cells in the pathogenesis of Multiple Sclerosis (MS) is incompletely understood. Here we define a possible role for B cells as myelin-specific antigen presenting cells (B-APCs) in MS. Peripheral blood B cells (PBBC) isolated from both MS patients and healthy controls (HC) were activated in vitro with either CD40L/IL-4 or a Class B CpG oligodeoxynucleotide (CpG ODN)/IL-2. Both activation techniques induced PBBCs to upregulate CD80 and HLA-DR, rendering them more efficient APCs than resting B cells. Although the CD40L/IL-4 B-APCs were highly effective in eliciting CNS-antigen specific proliferation by autologous T cells, CpG ODN/IL-2 stimulated B cells were not. Furthermore, CD40L/IL-4 B-APC induced responses by autologous CD4(+) T cells were susceptible to blocking with anti-HLA-DR antibody, suggesting that T cell responses were specific for antigen presentation by B-APC. CNS-antigen specific CD8(+) T cell proliferation was also blocked by HLA-DR, suggesting that CD8(+) proliferation is in part dependent on CD4(+) help.


Expression of Islet1 marks the sensory and neuronal lineages in the mammalian inner ear.
Several basic helix-loop-helix (bHLH) genes have been shown to be essential for the generation of the auditory sensory hair cells or the spiral ganglion (SG) neurons that innervate the hair cells in the cochlea, as well as a variety of cell types in the other nervous systems. However, it remains elusive what cellular context-dependent mechanisms confer the inner ear-specific neuronal or sensory competency/identities. We explored the possibility that one of the mechanisms responsible for generating cellular diversity in the nervous system through cooperative action of bHLH and LIM-homeodomain (LIM-HD) transcriptional factors might also contribute to the inner ear-specific sensory and/or neuronal competency. Here, we show that Islet1 (Isl1), a LIM-HD protein, is expressed early in the otocyst in the region that gives rise to both the auditory sensory organ, the organ of Corti, and SG neurons. Subsequently, the expression of Isl1 is maintained in SG neurons but is transitory in the sensory lineage. At embryonic day 12 (E12) in mice, the expression of Isl1 marks distinctively the ventral portion of the nascent cochlear epithelium encompassing the primordial organ of Corti. At E13, Isl1 is maintained at relatively high levels in the sensory primordium while down-regulated in the other regions of the cochlear duct. As the sensory epithelium starts to differentiate, it is down-regulated in the entire cochlear epithelium. The expression of Isl1 in the developing inner ear reveals an early and likely a common step in the development of both sensory and neuronal lineages of the inner ear, and suggests its potential role in the inner ear-specific sensory and neuronal cell development.


An Unusual Case of Acute Psychosis With Obsessive-Compulsive Features Following Arsenic Poisoning.
Arsenic exposure, particularly the chronic type, can lead to poisoning with manifestations presenting in multiple organ systems. However, acute psychosis is not a commonly described manifestation of arsenic exposure. In this report, we present the case of a patient who developed acute psychosis with hallucinations, disorganized thinking, and obsessive-compulsive symptoms following chronic occupational arsenic exposure. The patient was treated with the combination of an antipsychotic and an antidepressant and he responded well with significant improvement in both the acute psychosis and obsessive-compulsive symptoms. The authors concluded that patients can develop atypical symptoms, including acute psychosis, following arsenic poisoning. In the case described in this report, the patient also presented with a new onset of obsessive-compulsive symptoms. Given this rare manifestation of arsenic poisoning for which there is no clearly defined treatment regimen, this case suggests that the use of a combination of an antipsychotic and an antidepressant may be considered in the rare event of psychosis with obsessive-compulsive features following arsenic poisoning.


Patient reported burden of asthma on resource use and productivity across 11 countries in Europe.
Asthma affects 30 million people in Western Europe, leading to substantial burden on healthcare systems and economies. REcognise Asthma and LInk to Symptoms and Experience (REALISE™) was a large European survey across 11 countries assessing patient attitudes and behaviors towards their asthma. The present study utilizes REALISE™ data to understand resource use and absenteeism in asthma. Data were collected on absenteeism and healthcare resource use from 8000 asthma patients (aged 18-50 years) across the 11 countries. All data were patient reported. Odds ratios (ORs) were calculated against the country with the lowest proportion of respondents for hospitalization (as a proxy for lowest resource use). Patient characteristics were broadly similar across countries. However, self-reported asthma control status varied. More than 50% of respondents in most countries considered primary healthcare professionals (HCPs), i.e., general practitioners and nurses, the main HCP they see about their asthma. However, in some countries, specialists or nurses were considered the main HCP. Hospitalization was lowest amongst patients in the Netherlands. Resource use and productivity loss varied widely across the countries; ORs for hospitalization ranged from 1 in Sweden to 4 in Norway and for productivity loss from 0.6 in Sweden to 2.6 in Italy, compared with the Netherlands. This study quantified utilization of healthcare resources in asthma (number of visits of HCPs, hospitalization, and accident and emergency visits) as well as absenteeism and showed that differences exist across countries. The differences in primary care and specialist use suggest a possible difference in healthcare delivery across countries. Mundipharma International Limited, Cambridge, UK.


[Ceraxon (citicoline) in the treatment of the mild cognitive impairment syndrome].
The objective was to study efficacy and safety of ceraxon (citicoline) used perorally in dose 1000 mg daily in the treatment of cognitive disturbances in patients with amnesic type of mild cognitive impairment (MCI). Twenty patients, aged from 50 to 90 years, received ceraxon in dose 1000 mg twice a day during 90 days. The state of patients was assessed with a battery of scales and tests with the following statistical data analysis. The significant results on the clinical effect of the drug on cognitive disturbances and good tolerability of ceraxon have been obtained. The authors recommend ceraxon for long-term preventive treatment in high risk populations for Alzheimer's disease.


Insulin autoantibodies with high affinity to the bovine milk protein alpha casein.
Insulin autoantibodies (IAA) can appear in children within months of introducing solid foods to the diet and before clinical type 1 diabetes. The aim of this study was to determine whether infant dietary antigens could be immunizing agents of IAA. To this end, IAA binding to [(125) I]insulin was competed with food preparations and extracts of foods encountered in the infant diet (milk formulas, bovine milk, wheat flour, fowl meal). Bovine milk powder extracts inhibited IAA-positive samples from six of 53 children (age 0·3-14·0 years) participating in German prospective cohorts. Inhibition in these sera ranged from 23 to 100%. Competition was abolished when hydrolyzed milk powder was used. Competition with protein components of bovine milk found that two of the six milk-reactive sera were inhibited strongly by alpha- and beta-casein; none were inhibited by the milk proteins bovine serum albumin or lactoglobulins. The two casein-reactive sera had high affinity to alpha-casein (1·7×10(9) ; 3·1×10(9) l/mol), and lesser affinity to beta-casein (4·0×10(8) ; 7·0×10(7) l/mol) and insulin (2·6×10(8) ; 1·6×10(8) l/mol). No children with milk-reactive IAA developed autoantibodies to other islet autoantigens or diabetes (median follow-up 9·8 years). These results suggest that autoimmunity to insulin can occur infrequently via cross-reactivity to food proteins, but this form of IAA immunization does not appear to be associated with progression to diabetes.


Bioactive berry compounds-novel tools against human pathogens.
Berry fruits are rich sources of bioactive compounds, such as phenolics and organic acids, which have antimicrobial activities against human pathogens. Among different berries and berry phenolics, cranberry, cloudberry, raspberry, strawberry and bilberry especially possess clear antimicrobial effects against, e.g. Salmonella and Staphylococcus. Complex phenolic polymers, like ellagitannins, are strong antibacterial agents present in cloudberry and raspberry. Several mechanisms of action in the growth inhibition of bacteria are involved, such as destabilisation of cytoplasmic membrane, permeabilisation of plasma membrane, inhibition of extracellular microbial enzymes, direct actions on microbial metabolism and deprivation of the substrates required for microbial growth. Antimicrobial activity of berries may also be related to antiadherence of bacteria to epithelial cells, which is a prerequisite for colonisation and infection of many pathogens. Antimicrobial berry compounds may have important applications in the future as natural antimicrobial agents for food industry as well as for medicine. Some of the novel approaches are discussed.


Development, Psychometric Testing, and Use of a Disaster Nursing Competency Scale in Iran: A Mixed Methods Study.
Nurses play an important role during disaster response and recovery. Few tools are available to assess the relative competence of nurses in disaster. This study aimed to develop, test, and evaluate a disaster nursing competency scale. This was an exploratory mixed methods study. In the qualitative phase, 35 Iranian nurses and managers with experience in disaster relief were asked about the skills they need to respond effectively to a disaster event. From the qualitative data, a 50-item questionnaire was developed. In the quantitative phase, exploratory factor analysis was conducted with 400 samples followed by a test-retest reliability assessment with 30 nurses. The developed and validated instrument was applied to 200 nurses in Iran to assess their competency. From content analysis of the qualitative data, 5 main themes emerged. In the quantitative phase, exploratory factor analysis loaded 50 items into 4 domains named management, legal and ethical, specific personal, and technical competence. Use of the tool showed that nurses estimated their competence overall at 70.28%. Findings demonstrate the necessity for new domains in disaster nursing competencies; which may be applicable as a useful tool and guideline to improve nursing education and practice.


Folic acid-tethered poly(N-isopropylacrylamide)-phospholipid hybrid nanocarriers for targeted drug delivery.
A series of temperature-responsive lipopolymers have been synthesized by bioconjugating poly(N-isopropylacrylamide)n (n = 25, 40, 60) onto three different phospholipids by the combination of reversible addition fragmentation chain transfer polymerization and azide-alkyne click reactions. To achieve the active targeting of cancer cells, folic acid (FA) has also been tethered to the resulting hybrid materials. The doxorubicin (Dox) encapsulated uniform nanocarriers (150 nm in diameter) fabricated by the self-assembly of the lipopolymers display temperature responsive controlled release. The FA receptor-mediated delivery of Dox was then assessed using KB cell lines, and the anti-cancer activity was assessed by the blocking of folic acid receptors. The FA-tethered lipopolymers showing temperature-responsiveness are advantageous for the cell-specific release of Dox, potentiating their anti-cancer activity.


Meta-analysis of fenestrating versus reconstituting subtotal cholecystectomy in the management of difficult gallbladder.
To evaluate comparative outcomes of fenestrating and reconstituting subtotal cholecystectomy (STC) in patients with difficult gallbladder. A systematic search of electronic data sources and bibliographic reference lists were conducted. All comparative studies reporting outcomes of laparoscopic fenestrating and reconstituting STC were included and their risk of bias were assessed using ROBINS-I tool. Seven comparative studies were included enrolling 590 patients undergoing laparoscopic STC using either fenestrating (n = 353) or reconstituting (n = 237) approaches. Although fenestrating STC was associated with a significantly higher rate of bile leak (OR: 2.47, p = 0.007) compared to reconstituting STC, both approaches were comparable in terms of resolution of bile leak without (RD: -0.02, p = 0.86) or with (OR: 1.84, p = 0.40) postoperative ERCP. Moreover, there was no significant difference in development of bile duct injury (RD: -0.02, p = 0.16), need for postoperative ERCP (OR: 1.36, p = 0.49), wound infection (RD: 0.03, p = 0.27), re-operation (OR: 0.95, p = 0.95), gallbladder remnant cholecystitis (OR: 0.21, p = 0.09) or need for completion cholecystectomy (RD: 0.01, p = 0.59) between two groups. Fenestrating STC is associated with a higher risk of bile leak than the reconstructing technique. This issue can be mitigated by routine use of drains, delayed drain removal, and in selected cases endoscopic therapy. We encourage the fenestrating approach considering trends in improved short- and long-term outcomes.


Bone marrow deficiency of TRPC3 channel reduces early lesion burden and necrotic core of advanced plaques in a mouse model of atherosclerosis.
Macrophage apoptosis plays a determinant role in progression of atherosclerotic lesions. An important goal in atherosclerosis research is to identify new components of macrophage apoptosis that can eventually be exploited as molecular targets in strategies aimed at manipulating macrophage function in the lesion. In the previous work from our laboratory, we have shown that transient receptor potential canonical 3 (TRPC3) channel is an obligatory component of survival mechanisms in human and murine macrophages and that TRPC3-deficient non-polarized bone marrow-derived macrophages exhibit increased apoptosis, suggesting that in vivo TRPC3 might influence lesion development. In the present work, we used a bone marrow transplantation strategy as a first approach to examine the impact of macrophage deficiency of TRPC3 on early and advanced atherosclerotic lesions of Apoe(-/-) mice. After 3 weeks of high-fat diet, lesions in mice transplanted with bone marrow from Trpc3(-/-) donors were smaller and with reduced cellularity than controls. Advanced lesions from these mice exhibited reduced necrotic core, less apoptotic macrophages, and increased collagen content and cap thickness. In vitro, TRPC3-deficient macrophages polarized to the M1 phenotype showed reduced apoptosis, whereas both M1 and M2 macrophages had increased efferocytic capacity. Bone marrow deficiency of TRPC3 has a dual beneficial effect on lesion progression by reducing cellularity at early stages and necrosis in the advanced plaques. Our findings represent the first evidence for a role of a member of the TRPC family of cation channels in mechanisms associated with atherosclerosis.


Epidemiological Study of Mortality Among Workers Exposed to Tritium in France.
Workers of the Commissariat for Atomic Energy and Alternative Energy (CEA) may be potentially exposed to tritium over long periods. We aimed to assess the effect of tritium exposure on mortality in a cohort of employees followed by radiotoxicological monitoring. A total of 1,746 employees who worked for at least six months at one of three CEA centers were included between 1962 and 2011 (median follow-up 29.6 years). The cumulative dose of tritium was based on the quantification of tritium present from urinary excretion monitoring data from the beginning of occupational exposure to the end of such exposure or December 2011. Mortality was first compared to that in the French population using the standardized mortality ratio (SMR). Then, mortality risk ratios (RRs) per category of cumulative dose of tritium were estimated using categorical Poisson models adjusted for age at the onset of exposure, age, calendar period, sex, smoking, employment status, CEA center, and taking into account the number of person-years. The main causes of mortality were tumors (48%) and cardiovascular diseases (20%). The comparison of mortality within the cohort to that in the French population highlighted a lower rate for all-cause mortality and that due to cancer, related to the healthy worker effect bias. The regression model showed no effect of cumulative dose on all-cause mortality. The risk of death for most malignancies was positive, but not significant for the higher classes of doses relative to the reference class. The highest risk (not significant) was present for the class of higher doses for tumors of the larynx, trachea, bronchi and lung. The risk was significant for the higher doses for tumors of the pancreas and bladder (based on a limited number of cases: five and six deaths, respectively). Significantly more smokers died from tumors of the respiratory system than non-smokers, as expected. We were unable to show an effect of cumulative tritium dose due to the small size of the cohort and the low exposure level. However, our study underlines the need to continue following tritium-exposed workers and conducting multicenter studies.


Biogeographic consequences of nutrient enrichment for plant-herbivore interactions in coastal wetlands.
A major challenge in ecology is to understand broadscale trends in the impact of environmental change. We provide the first integrative analysis of the effects of eutrophication on plants, herbivores, and their interactions in coastal wetlands across latitudes. We show that fertilisation strongly increases herbivory in salt marshes, but not in mangroves, and that this effect increases with increasing latitude in salt marshes. We further show that stronger nutrient effects on plant nitrogen concentration at higher latitudes is the mechanism likely underlying this pattern. This biogeographic variation in nutrient effects on plant-herbivore interactions has consequences for vegetation, with those at higher latitudes being more vulnerable to consumer pressure fuelled by eutrophication. Our work provides a novel, mechanistic understanding of how eutrophication affects plant-herbivore systems predictably across broad latitudinal gradients, and highlights the power of incorporating biogeography into understanding large-scale variability in the impacts of environmental change.


The maintenance of blood colloid: passage of stored gum acacia from the cells to the circulation after plasmapheresis.
Removal of blood plasma by plasmapheresis from dogs made hypoproteinemic by injections of gum acacia over long periods of time, has resulted in the removal of more gum acacia than was originally present in the plasma. Gum acacia injections had been discontinued previous to the start of the experiments, and hence it must be concluded that the excess amounts of acacia were derived from deposits in the various organs. These observations verify the previous suggestion that the increase in blood acacia in the hypoproteinemic dog on a low protein diet is due to the transfer of acacia to the blood from its sites of deposit in the body. The experiments further suggest that the colloid content of the blood stream is maintained at the expense of tissue colloids, and support the idea that colloidal substances may pass in and out of cells.


Endoscopic characteristics and levodopa responsiveness of swallowing function in progressive supranuclear palsy.
Dysphagia is a frequent and early symptom in progressive supranuclear palsy (PSP) predisposing patients to aspiration pneumonia. Fiberoptic endoscopic evaluation of swallowing (FEES) has emerged as a valuable apparative tool for objective evaluation of neurogenic dysphagia. This is the first study using FEES to investigate the nature of swallowing impairment in PSP. Eighteen consecutive PSP patients (mean age 69.7 +/- 9.0 years) were included. The salient findings of FEES in PSP patients were compared with those of 15 patients with Parkinson's disease (PD). In 7 PSP patients, a standardized FEES protocol was performed to explore levodopa (L-dopa) responsiveness of dysphagia. Most frequent abnormalities detected by FEES were bolus leakage, delayed swallowing reflex, and residues in valleculae and piriformes. Aspiration events with at least one food consistency occurred in nearly 30% of PSP patients. Significant pharyngeal saliva pooling was observed in 4 PSP patients. We found no difference of salient endoscopic findings between PSP and PD patients. Endoscopic dysphagia severity in PSP correlated positively with disease duration, clinical disability, and cognitive impairment. No correlation was found with dysarthria severity. In early PSP patients, swallowing dysfunction was solely characterized by liquid leakage with the risk of predeglutitive aspiration during the oral phase of swallowing. Two PSP patients showed relevant improvement of swallowing function after L-dopa challenge. Chin tuck-maneuver, hard swallow, and modification of food consistency were identified as the most effective therapeutic interventions. In conclusion, FEES assessment can deliver important findings for the diagnosis and refined therapy of dysphagia in PSP patients.


Spectroscopic and Computational Studies of Spin States of Iron(IV) Nitrido and Imido Complexes.
High-oxidation-state metal complexes with multiply bonded ligands are of great interest for both their reactivity as well as their fundamental bonding properties. This paper reports a combined spectroscopic and theoretical investigation into the effect of the apical multiply bonded ligand on the spin-state preferences of threefold symmetric iron(IV) complexes with tris(carbene) donor ligands. Specifically, singlet (S = 0) nitrido [{PhB(ImR)3}FeN], R = tBu (1), Mes (mesityl, 2) and the related triplet (S = 1) imido complexes, [{PhB(ImR)3}Fe(NR')]+, R = Mes, R' = 1-adamantyl (3), tBu (4), were investigated by electronic absorption and Mössbauer effect spectroscopies. For comparison, two other Fe(IV) nitrido complexes, [(TIMENAr)FeN]+ (TIMENAr = tris[2-(3-aryl-imidazol-2-ylidene)ethyl]amine; Ar = Xyl (xylyl), Mes), were investigated by 57Fe Mössbauer spectroscopy, including applied-field measurements. The paramagnetic imido complexes 3 and 4 were also studied by magnetic susceptibility measurements (for 3) and paramagnetic resonance spectroscopy: high-frequency and -field electron paramagnetic resonance (for 3 and 4) and frequency-domain Fourier-transform (FD-FT) terahertz electron paramagnetic resonance (for 3), which reveal their zero-field splitting parameters. Experimentally correlated theoretical studies comprising ligand-field theory and quantum chemical theory, the latter including both density functional theory and ab initio methods, reveal the key role played by the Fe 3dz2 (a1) orbital in these systems: the nature of its interaction with the nitrido or imido ligand dictates the spin-state preference of the complex. The ability to tune the spin state through the energy and nature of a single orbital has general relevance to the factors controlling spin states in complexes with applicability as single molecule devices.


Roles of Ubiquitination and SUMOylation in DNA Damage Response.
Ubiquitin and ubiquitin-like modifiers, such as SUMO, exert distinct physiological functions by conjugating to protein substrates. Ubiquitination or SUMOylation of protein substrates determine the fate of modified proteins, including proteasomal degradation, cellular re-localization, alternations in binding partners and serving as a protein-binding platform, in a ubiquitin or SUMO linkage-dependent manner. DNA damage occurs constantly in living organisms but is also repaired by distinct tightly controlled mechanisms including homologous recombination, non-homologous end joining, inter-strand crosslink repair, nucleotide excision repair and base excision repair. On sensing damaged DNA, a ubiquitination/SUMOylation landscape is established to recruit DNA damage repair factors. Meanwhile, misloaded and mission-completed repair factors will be turned over by ubiquitin or SUMO modifications as well. These ubiquitination and SUMOylation events are tightly controlled by both E3 ubiquitin/SUMO ligases and deubiquitinases/deSUMOylases. In this review, we will summarize identified ubiquitin and SUMO-related modifications and their function in distinct DNA damage repair pathways, and provide evidence for responsible E3 ligases, deubiquitinases, SUMOylases and deSUMOylases in these processes. Given that genome instability leads to human disorders including cancer, understanding detailed molecular mechanisms for ubiquitin and SUMO-related regulations in DNA damage response may provide novel insights into therapeutic modalities to treat human diseases associated with deregulated DNA damage response.


Tracheal extubation in children with difficult airways: a descriptive cohort analysis.
Tracheal extubation in children with difficult airways may be associated with an increased risk of perioperative adverse events. The aim of this study was to describe the exubation techniques used/ success rates/ adverse events related to tracheal extubation practices in children with difficult airways. A retrospective analysis of tracheal extubation practices in the difficult airway population over a 78-month period was performed. Difficult airway was defined as a Cormack and Lehane Grade 3 view or greater, and/or tracheal intubation requiring ≥ 3 attempts, and/or the need for an alternate device to direct laryngoscopy for successful tracheal intubation, and/or difficult mask ventilation. Reasons for difficult airway, demographic/surgical data, technique(s) for tracheal extubation, success/failure of tracheal extubation, and adverse events were recorded. A failed tracheal extubation was defined as any adverse event related to the airway occurring within 6 h of extubation requiring reintubation. A total of 519 patients were reported to have a difficult airway during this study period in a tertiary care pediatric center. Of these, 137 patients (26%) met inclusion criteria. Tracheal extubation was successfully performed in 130 patients (95%). The majority of tracheal exubations were performed without the use of additional airway adjuncts straight onto anesthesia face mask (121/137; 88%). Extubation failure occurred in seven cases (5%). Among the failed extubations, 6/7 children (85%) had evidence of severe upper airway obstruction and were <10 kg in weight. Of these children, one child required emergency tracheostomy, and two children (one with tracheal stenosis and other with spinal muscular atrophy) suffered from hypoxemic cardiac arrest and anoxic brain damage, respectively, and eventually died. In the studied population of children with difficult airways handled in a tertiary center environment, the majority of tracheal extubations could be performed without the use of airway adjuncts. In a minority of patients, tracheal extubation was associated with severe adverse outcomes.


An efficient and enantioselective approach to the azaspirocyclic core of alkaloids: formal synthesis of halichlorine and pinnaic acid.
A novel, highly stereoselective synthesis of an azaspirocyclic core, which contains four stereogenic carbons consistent with structures of natural halichlorine and pinnaic acid, is presented. Lipase PS-catalyzed selective acylation, asymmetric methylation on the alpha-methylene of the bicyclic lactone, and an asymmetric Michael addition of the tertiary nitro cyclopentane were concisely used to conquer the challenging problem of successfully constructing the C9 quarternary carbon center with complete stereocontrol. The spiropiperidine ring was formed by reduction of the delta-nitroketone, intramolecular condensation, and then highly stereoselective reduction of the cyclic nitrone with NaBH(4). This spirocyclic core is a key intermediate in Danishefsky's synthesis of pinnaic acid and halichlorine.


Self-purificatory Ganga water facilitates death of pathogenic Escherichia coli O157:H7.
Concern over the prevalence of active pharmaceutical agents and subsequent occurrence of antimicrobial resistance in the environment is increasing. Incorruptible ability of Ganga water was evaluated using fresh, 8-year-old, and 16-year-old Ganga water samples spiked with pathogenic Escherichia coli serotype O157:H7. Survival of E. coli O157:H7 over the course of the experiment was 3, 7, and 15 days for fresh, 8-year-old, and 16-year-old Ganga waters, respectively. On the contrary, in Milli Q water the decline in viable count of E. coli O157:H7 up to 30 days was only 2 log units. Survival of E. coli O157:H7 was greater in boiled water compared with water after passage through a 0.2-microm-pore-size membrane filter, indicating involvement of heat-labile agents influencing survival of E. coli O157:H7 in Ganga water, which seems to indicate the role of antimicrobial peptides. Functional diversity of Ganga water's native microbial community structure as assessed with Biolog Eco plates was not affected even in the presence of a 5-fold log units higher pathogenic load of E. coli O157:H7. These findings suggest that Ganga water has certain novel antimicrobial attributes, besides its remarkable fluidity, which may provide a much-needed basis for the development of new antimicrobial compounds.


The role of digital health in emergency medicine education.
Despite the need for a well-trained emergency medicine (EM) workforce in India, there are still many shortcomings. The COVID-19 pandemic has brought to the limelight the critical importance of digital health data, tools, technologies, and services in monitoring and combating adverse health effects of the pandemic. On the other hand, relevant health care stakeholders have often been underprepared to embrace digital health solutions wholeheartedly and rise to the pandemic challenge. The Global Strategy on Digital Health 2020-2025 clearly states how relevant digital health interventions (DHI) would enhance the access and improve the health outcomes, if used judiciously. Currently, India's G20 Presidency health priorities are focussed on health emergencies prevention, preparedness, and response, by incorporating digital health at its core. Keeping these policy directives in mind, health professional educators should actively incorporate DHI as an integral part of health professional education for the domain of EM. To attain an optimal growth of EM as an academic discipline in India, the curriculum has to be made up to date, incorporating the latest DHIs. The PG curriculum committees of both the National Medical Commission and the National Board of Examination must form a consensus for implementing such curriculum throughout India. The modern-day emergency room (ER) or casualty may soon have AI-enabled robots doing a lot of repetitive and precise activities to save lives. Healthcare professionals working in the ER have to be aware and efficient in handling these robots and other clinical decision support systems well.


The value of LYM-1 cells for examining vacuole formation and loss of cell viability induced by culture supernates of Helicobacter pylori.
Some strains of Helicobacter pylori are known to produce an extracellular cytotoxin that causes vacuolation in cultured mammalian cells. Screening for such strains makes use of HeLa cells which may not be sensitive enough to detect minimal changes. The aim of this study was to develop a more sensitive cell line. Vacuole formation was examined in HeLa cells, as well as four other cell lines established in this laboratory by ammonium chloride induction. Among five cell lines tested, LYM-1 cells were most sensitive for the detection of intracellular vacuolation with this agent. Loss of cell viability of LYM-1 and HeLa cells induced by H. pylori culture supernates was also examined: LYM-1 were more sensitive than HeLa cells. Cell death was not always accompanied by vacuole formation. This suggests that the mechanism whereby cell death occurs must be different from that for vacuole formation. LYM-1 cells may be useful when measuring vacuole formation and cell death of the cultured cells induced by culture supernates of clinical isolates of H. pylori.


Patient Safety Culture and the Ability to Improve: A Proof of Concept Study on Hand Hygiene.
OBJECTIVE To investigate whether the safety culture of a hospital unit is associated with the ability to improve. DESIGN Qualitative investigation of safety culture on hospital units following a before-and-after trial on hand hygiene. SETTING VU University Medical Center, a tertiary-care hospital in the Netherlands. METHODS With support from hospital management, we implemented a hospital-wide program to improve compliance. Over 2 years, compliance was measured through direct observation, twice before, and 4 times after interventions. We analyzed changes in compliance from baseline, and selected units to evaluate safety culture using a positive deviance approach: the hospital unit with the highest hand hygiene compliance and 2 units that showed significant improvement (21% and 16%, respectively) were selected as high performing. Another 2 units showed no improvement and were selected as low performing. A blinded, independent observer conducted interviews with unit management, physicians, and nurses, based on the Hospital Survey on Patient Safety Culture. Safety culture was categorized as pathological (lowest level), reactive, bureaucratic, proactive, or generative (highest level). RESULTS Overall, 3 units showed a proactive or generative safety culture and 2 units had bureaucratic or pathological safety cultures. When comparing compliance and interview results, high-performing units showed high levels of safety culture, while low-performing units showed low levels of safety culture. CONCLUSIONS Safety culture is associated with the ability to improve hand hygiene. Interventions may not be effective when applied in units with low levels of safety culture. Although additional research is needed to corroborate our findings, the safety culture on a unit can benefit from enhancement strategies such as team-building exercises. Strengthening the safety culture before implementing interventions could aid improvement and prevent nonproductive interventions. Infect Control Hosp Epidemiol 2017;38:1277-1283.


Video games are exciting: a European study of video game-induced seizures and epilepsy.
Video game seizures have been reported in photosensitive and non-photosensitive patients with epilepsy. The game Super Mario World, has led to many cases of first seizures. We examined whether this game was indeed more provocative than other programs and whether playing the game added to this effect. We prospectively investigated 352 patients in four European cities, using a standard protocol including testing of a variety of visual stimuli. We correlated historical data on provocative factors in daily life with electroencephalographic laboratory findings. The video game, Super Mario World proved more epileptogenic than standard TV programs and as provocative as programs with flashing lights and patterns. Most striking was the fact that video game-viewing and-playing on the 50 and 100 Hz TV was significantly more provocative than viewing the standard program (P < 0.001, P < 0.05 respectively). Playing the video game Mario World on a 50 Hz TV, appeared to be significantly more provocative than playing this game on the 100 Hz TV (P < 0.001). Of 163 patients with a history of TV-, VG- or CG-seizures, 85% of them showed epileptiform discharges in response to photic stimulation, 44% to patterns, 59% to 50 Hz TV and 29% to 100 Hz TV. Children and adolescents with a history of video game seizures are, in the vast majority, photosensitive and should be investigated with standardised photic stimulation. Games and programs with bright background or flashing images are specifically provocative. Playing a video game on a 100 Hz TV is less provocative [published with videosequences].


Effects of gestational undernutrition, stress and diazepam treatment on spatial discrimination learning and retention in young rats.
Effects of prenatal undernutrition, stress and diazepam treatment on learning acquisition, and subsequent retention of a spatial discrimination task was assessed in the offsprings. Undernutrition of the dams was induced by restricting food intake to half, throughout the period of gestation, whereas footshock stress and diazepam (0.5 mg/kg, ip) treatment was given from day 13 to 20 of gestation, this being the critical period for neural development in this species. The pups born were subjected to spatial discrimination learning, and retention of the learning acquisition after an interval of one week, in a single unit black/white T-maze, at 8-9 weeks of age. The results indicate that prenatal undernutrition induces significant learning and retention deficits in the offspring, whereas the effect of prenatal stress was limited to only deficit in learning acquisition. Prenatally administered diazepam induced significant deficits in learning acquisition and subsequent retention of the discrimination task in pups culled from normally nourished dams. However, offsprings from diazepam administered undernourished dams exhibited less marked cognitive deficits, which may be attributable to the altered emotional reactivity of pups born to undernourished mothers. Prenatally administered diazepam also induced differential effects in stressed and non-stressed dam offsprings, though the effects were statistically insignificant. The results suggest that prenatal insults, in the form of undernourishment, stress and anxiolytic drugs, leave a lasting imprint on cognitive behaviour of the offspring. The final effect on this behaviour may be determined by the co-existence of these prenatal factors, particularly at a time when the foetus is vulnerable because of neural development and differentiation.


Dietary and Health Correlates of Sweetened Beverage Intake: Sources of Variability in the National Health and Nutrition Examination Survey (NHANES).
Recent studies using data from the National Health and Nutrition Examination Survey (NHANES) have used inconsistent approaches to identify and categorize beverages, especially those containing low-calorie sweeteners (LCS), also referred to as low-calorie sweetened beverages (LCSBs). Herein, we investigate the approaches used to identify and categorize LCSBs in recent analyses of NHANES data. We reviewed published studies examining LCS consumption in relation to dietary and health outcomes and extracted the methods used to categorize LCS as reported by the authors of each study. We then examined the extent to which these approaches reliably identified LCSBs using the Internet Archive Wayback Machine to examine beverage ingredients lists across three NHANES cycles (2011-2016). None of the four general strategies used appeared to include all LCSBs while also excluding all beverages that did not contain LCS. In some cases, the type of sweetener in the beverage consumed could not be clearly determined; we found 9, 16, and 18 of such "mixed" beverage identifiers in the periods 2011-2012, 2013-2014, and 2015-2016, respectively. Then, to illustrate how heterogeneity in beverage categorization may impact the outcomes of published analyses, we compared results of a previously published analysis with outcomes when "mixed" beverages were grouped either all as LCSBs or all as sugary beverages. Our results suggest that caution is warranted in design and interpretation of studies using NHANES data to examine dietary and health correlates of sweetened beverage intake.


[Cardiovascular calcification inhibitors].
Vascular calcification is a marker of cardiovascular risk increase. Age and specific disease such as diabetes or chronic kidney disease are important factors for calcification genesis. Vascular calcification process is a complex phenomenon, involving several activators and inhibitors factors. Indeed, recent works related to in vitro and in vivo experimental studies have led to a better understanding of calcification process and identification of molecules able to modulate this system. This revue will summarize some of these molecules with a particular interest of those with therapeutic relevance. We will present: i) calcium sensing receptor and its modulation by cinacalcet; ii) pyrophosphate supplementation; iii) fetuin A and overall propensity serum test for calcification and finally; iv) matrix-Gla-protein and the use of vitamin K to prevent vascular calcification progression.


Federal funding of nursing research by the National Institutes of Health (NIH): 1993 to 2017.
There has been a dramatic increase in the number of doctoral programs (PhD and DNP) that prepare nurse research scientists and advanced practitioners since establishment of the National Institute of Nursing Research (NINR) at the National Institutes of Health (NIH) in 1985. The purpose of this report is to examine the historical context of federal research funding to schools/colleges of nursing to determine if the NINR/NIH budget is adequate. Data were extracted from the NIH RePORT/ER database from 1993 to 2017. Additional data were obtained from the American Association of Colleges of Nursing. A return on investment analysis for four landmark nursing studies is included. The percent of the NINR budget awarded to schools/colleges of nursing peaked in 2005; since 2011, more funding to schools/colleges of nursing was received from all other NIH institutes combined, compared to NINR. The return on investment for four nursing research studies, ranged from $1:$202 to $1:$1,206, and far exceeds the Standard and Poor's 500 Index (S&P 500) of 10%. Federal funding of nursing research is inadequate and a chokepoint relative to the number of doctoral programs. We suggest the NINR budget would need to increase at least fivefold to over $763 million to adequately fund nursing science. The impact of inadequate funding on the discipline is discussed.


Legume consumption and severe depressed mood, the modifying roles of gender and menopausal status.
Legumes have been recommended as staple foods in the anticipation of disease prevention. However, the scientific evidence of their benefits, particularly on mental well-being, remains preliminary. We longitudinally assessed the association between legume consumption and the risk of severe depressed mood (SDM) among a national cohort. The study included adults aged 25-74 years who were examined in 1971-1975 as a part of the National Health and Nutrition Examination Survey. Legume consumption at baseline was obtained from a 3-month FFQ and categorised as infrequent (<1 time/week), moderate (1-2 times/week) and frequent (> or = 3 times/week). SDM was defined as Center for Epidemiologic Studies Depression Scale score > or = 22 or taking anti-depression medication after an average of 10.6 years of follow-up (from 8.0 to 12.5 years). Among women, the proportion of individuals with SDM was 17.75 %. For premenopausal women (n 1778), a significant linear trend of deleterious effect from legume consumption was observed (P for trend = 0.0148). The relative risks (RR) for infrequent, moderate and frequent consumptions were 1 (reference), 1.24 (95 % CI = 0.91, 1.70) and 1.75 (1.12, 2.75), respectively. However, moderate consumption showed a significant protective effect (RR = 0.52 (0.27, 1.00)) among women undergoing the menopausal transition (n 454). No association was obtained from either postmenopausal women (n 601) or men (n 2036). These findings suggest that gender and menopausal status were effect modifiers of the association between legume consumption and SDM. Detrimental effects of frequent consumption of legumes may exist among premenopausal women; moderate consumption, however, may protect perimenopausal women against SDM.


Soft Surfaces for Fast Characterization and Positioning of Scanning Electrochemical Microscopy Nanoelectrode Tips.
The testing of nanoelectrode tips for scanning electrochemical microscopy (SECM) is a slow and cumbersome task that often results in untimely electrode breakage due to crashing against a substrate. Here, we evaluated approach curves of nano- and microelectrodes to soft surfaces using SECM for a rapid and more convenient characterization and positioning protocol. Soft surfaces consisted of either a submerged argon bubble or a thin polydimethylsiloxane (PDMS) layer. While approach curves to Ar bubbles in the presence of a surfactant were promising for the characterization of microelectrode tips, their performance with nanoelectrodes was deficient. In contrast, approach curves to PDMS films allowed the rapid positioning of nanoelectrodes as small as 30 nm radius at speeds up to 5 μm/s without the risk of breakage. The nanoelectrodes were able to approach the polymer films multiple times without affecting their electrochemical performance. Furthermore, using a half-coated substrate with PDMS, nanoelectrodes could be retracted and positioned very close to the bare, hard substrate for characterization with traditional approach curves. We estimate time savings on tip characterization/positioning on the order of 10- to 100-fold. This simple procedure is easily implemented without the requirement of additional devices supplementing existing commercial SECM instruments.


Inhibitor of differentiation-3 mediates high fat diet-induced visceral fat expansion.
Inhibitor of differentiation-3 (Id3) has been implicated in promoting angiogenesis, a key determinant of high-fat diet (HFD)-induced visceral adiposity. Yet the role of Id3 in HFD-induced angiogenesis and visceral adipose expansion is unknown. Id3(-/-) mice demonstrated a significant attenuation of HFD-induced visceral fat depot expansion compared to wild type littermate controls. Importantly, unlike other Id proteins, loss of Id3 did not affect adipose depot size in young mice fed chow diet or differentiation of adipocytes in vitro or in vivo. Contrast enhanced ultrasound revealed a significant attenuation of visceral fat microvascular blood volume in HFD-fed mice null for Id3 compared to wild type controls. HFD induced Id3 and VEGFA expression in the visceral stromal vascular fraction and Id3(-/-) mice had significantly lower levels of VEGFA protein in visceral adipose tissue compared to wild type. Furthermore, HFD-induced VEGFA expression in visceral adipose tissue was completely abolished by loss of Id3. Consistent with this effect, Id3 abolished E12-mediated repression of VEGFA promoter activity. Results identify Id3 as an important regulator of HFD-induced visceral adipose VEGFA expression, microvascular blood volume, and depot expansion. Inhibition of Id3 may have potential as a therapeutic strategy to limit visceral adiposity.


Cognitive enhancement drug use among resident physicians: Prevalence and motivations for use - results from a survey.
Background: Non-medical use of prescription drugs for the enhancement of cognitive functioning has gained popularity in recent years, especially among young educated adults. To our knowledge, no previous study investigated this phenomenon among resident physicians.Objective: To analyze cognitive enhancement drugs use motivations and patterns among resident physicians.Methods: A survey and statistical analysis regarding the use of drugs traditionally prescribed for the treatment of Attention Deficit Hyperactivity Disorder: stimulants, amphetamines and modafinil.Participants: 1,453 residents who took their written residency exam in the summer of 2017. The response rate was 32.3%.Results: 28.1% of responders reported past use, with 73.67% of them reporting use without a related medical diagnosis. Almost half of the users (47.1%) acquired the drug with a prescription, but without a diagnosis of a related medical disorder. The first use was predominantly during residency (54.3%), with 45% reporting it as related to the residency exam.Factors found to positively impact non-medical use include: declaring undiagnosed Attention Deficit Hyperactivity Disorder, fear of failing the exam, a belief that more than 30% of other examinees take cognitive enhancements drugs, and a learning disability diagnosis. Self-reports of being a competitive person and being a parent, were negatively correlated with non-medical use.Conclusions: The use of drugs that are taken traditionally for the treatment of Attention Deficit Hyperactivity Disorder is common among resident physicians, both with and without related medical indication. Interestingly, factors associated with the fear of being "left behind" increase non-medical use and not the desire to succeed.


Ottowia beijingensis sp. nov., isolated from coking wastewater activated sludge, and emended description of the genus Ottowia.
A taxonomic study was carried out on strain GCS-AN-3(T), which was isolated from a phenol-degrading consortium enriched from coking wastewater activated sludge of Beijing Shougang Company Limited during the screening of phenol-degrading bacteria. Cells of strain GCS-AN-3(T) were Gram-stain-negative, short rods, and oxidase-/catalase-positive. Growth was observed at salinities from 0 to 2.5 % and at temperatures from 10 to 37 °C. 16S rRNA gene sequence analysis showed that strain GCS-AN-3(T) was most closely related to Ottowia pentelensis DSM 21699(T) (96.2 %). The principal fatty acids were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), C16 : 0, summed feature 8 (C18 : 1ω7c/C18 : 1ω6c) and cyclo C17 : 0. The major respiratory quinone was Q-8. The polar lipids comprised phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine and an unknown phospholipid. The G+C content of the genomic DNA was 67.6 mol%. Thiosulfate could be utilized as co-substrate for aerobic growth and was oxidized to sulfate. On the basis of phenotypic, chemotaxonomic and molecular data, strain GCS-AN-3(T) is considered to represent a novel species of the genus Ottowia, for which the name Ottowia beijingensis sp. nov. is proposed (type strain GCS-AN-3(T) = LMG 27179(T) = CGMCC 1.12324(T) = MCCC 1A01410(T)). An emended description of the genus Ottowia is also proposed.


Somatic copy number gains of α-synuclein (SNCA) in Parkinson's disease and multiple system atrophy brains.
The α-synuclein protein, encoded by SNCA, has a key role in the pathogenesis of Parkinson's disease and other synucleinopathies. Although usually sporadic, Parkinson's disease can result from inherited copy number variants in SNCA and other genes. We have hypothesized a role of somatic SNCA mutations, leading to mosaicism, in sporadic synucleinopathies. The evidence for mosaicism in healthy and diseased brain is increasing rapidly, with somatic copy number gains of APP reported in Alzheimer's brain. Here we demonstrate somatic SNCA copy number gains in synucleinopathies (Parkinson's disease and multiple system atrophy), focusing on substantia nigra. We selected sporadic cases with relatively young onset or short disease duration, and first excluded high level copy number variant mosaicism by DNA analysis using digital PCR for SNCA, and/or customized array comparative genomic hybridization. To detect low level SNCA copy number variant mosaicism, we used fluorescent in situ hybridization with oligonucleotide custom-designed probes for SNCA, validated on brain and fibroblasts with known copy number variants. We determined SNCA copy number in nigral dopaminergic neurons and other cells in frozen nigra sections from 40 cases with Parkinson's disease and five with multiple system atrophy, and 25 controls, in a blinded fashion. Parkinson's disease cases were significantly more likely than controls to have any SNCA gains in dopaminergic neurons (P = 0.0036), and overall (P = 0.0052). The average proportion of dopaminergic neurons with gains in each nigra was significantly higher in Parkinson's disease than controls (0.78% versus 0.45%; P = 0.017). There was a negative correlation between the proportion of dopaminergic neurons with gains and onset age in Parkinson's disease (P = 0.013), but not with disease duration, or age of death in cases or controls. Cases with tremor at onset were less likely to have gains (P = 0.035). All multiple system atrophy cases had gains, and the highest levels in dopaminergic neurons were in two of these cases (2.76%, 2.48%). We performed selective validation with different probes after dye swapping. All three control probes used showed minimal or no gains (≤0.1% in dopaminergic neurons). We also found occasional SNCA gains in frontal neurons of cases with Parkinson's disease, and the putamen of one multiple system atrophy case. We present evidence of somatic SNCA gains in brain, more commonly in nigral dopaminergic neurons of Parkinson's disease than controls, negatively correlated with onset age, and possibly commonest in some multiple system atrophy cases. Somatic SNCA gains may be a risk factor for sporadic synucleinopathies, or a result of the disease process.10.1093/brain/awy157_video1awy157media15813519976001.


Risk factors for extremely long length-of-stay among pediatric emergency patients.
Over time, we observed more visits in our pediatric emergency department with length-of-stay (LOS) of more than 10 hours, whereas our mean LOS was approximately 3 hours. We sought to characterize factors associated with this extremely long LOS. Eighty-one visits with LOS more than 10 hours were identified from January 1, 2001, to June 30, 2003. In this retrospective study, we compared these cases with 405 randomly selected age-matched controls with LOS less than 10 hours (5 controls per case). The groups were similar for sex, visit month, arrival mode, and level of training of the supervising physician. Cases more frequently arrived during night shifts (30% vs 13%) and had laboratory tests (93% vs 32%), radiological studies (83% vs 34%), procedures (28% vs 15%), sedations (24% vs 4%), subspecialty consultations (84% vs 20%), chief complaints of abdominal pain (42% vs 6%) and diagnoses of appendicitis (10% vs 1%), and had a greater hospitalization rate (67 vs 19%). Although more cases involved white patients (57% vs 31%), race was not associated with LOS more than 10 hours in adjusted analysis. In multivariable analysis, longer waiting time (odds ratio [OR], 1.013; 95% confidence interval [CI], 1.007-1.019), night shift arrival (OR, 5.0; 95% CI, 1.9-12.8), higher triage acuity (lowest acuity: OR, 0.003; 95% CI, 0.0-0.286), radiology study other than radiographs (OR, 18.0; 95% CI, 7.5-43.1), and subspecialty consultation (OR, 7.6; 95% CI, 3.2-18.3) were associated with LOS more than 10 hours. In our pediatric emergency department, risk factors for LOS more than 10 hours included longer waiting time, night shift arrivals, high triage acuity, radiology studies, and subspecialty consultations. These factors may also be important considerations for quality improvement initiatives at other institutions.


Incidence of the involvement of the cardiovascular system in HIV infection.
The epidemiology of cardiac complications related to HIV, including cardiomyopathy, increased left ventricular mass, myocarditis, pericardial effusion, endocarditis, and malignancy, are discussed. An increased number of HIV-infected individuals may present with cardiac complications in the next decade as chronic viral infection, co-infections, drug therapy, and immunosuppression affect the heart. Understanding the nature and course of cardiac illness related to HIV infection may allow appropriate monitoring, early intervention and therapy, and will provide a baseline to evaluate the effects of new therapeutic regimens such as highly active antiretroviral therapy on the cardiovascular system.


Eliminating partial-transformation products and mitigating residual toxicity of amoxicillin through intimately coupled photocatalysis and biodegradation.
Intimately coupled photocatalysis and biodegradation (ICPB) is a promising technology for treating wastewater containing antibiotics. While past work has documented the benefits of ICPB for removing and mineralizing antibiotics, its impacts on mitigating biotoxicity from products has not been studied. We fabricated an ICPB carrier by coating Ag-doped TiO2 on the outer skeleton of sponge carriers and allowing biofilm to grow in the internal macro-pores. We used amoxicillin (C16H19N3O5S) as the model antibiotic. The amoxicillin-removal rate contents with ICPB was greater by 40% vs. photocatalysis and 65% vs. biodegradation, based on the first-order kinetic simulation. While mineralization of amoxicillin was minimal for photocatalysis or biodegradation alone, it was ∼35% with ICPB. Photocatalysis alone led to accumulation of C14H21N3O2S; biodegradation alone resulted in accumulation of C14H21N3O3, C16H18N2O4S, and C15H21N3O3; but they were negligible after ICPB. As a result, ICPB reduced toxicity impacts measured by Staphylococcus aureas growth, Daphnia magna mobility, and teratogenicity to Zebrafish embryos. In contrast, photocatalysis alone increased each of the toxicity effects. In sum, ICPB gave greater removal and mineralization of amoxicillin, and it mitigated biotoxicity from treatment products.


Prevalence of cerebral microhemorrhages in amateur boxers as detected by 3T MR imaging.
The evidence for traumatic brain injury in amateur boxers is controversial. Hypothetically, sudden acceleration of the head due to the impact of the blow during the boxing fight might result in diffuse axonal injury or contusion. We wanted to determine whether cerebral microhemorrhages occur more often in amateur boxers than in nonboxers. In 42 male, classical amateur boxers and in 37 healthy, nonboxing male volunteers we performed cranial MR imaging at 3T. The study protocol included a transverse dual spin-echo MR imaging sequence, a 3D sagittal magnetization-prepared rapid acquisition of gradient echo sequence, a coronal T2*-weighted sequence, and an axial time-of-flight MR angiography sequence. MR imaging data were made anonymous before 2 neuroradiologists independently evaluated the images. In addition, the following risk factors were assessed: total numbers of fights and knockouts, weight division, and duration of boxing. We compared the group proportions of microhemorrhages with Fisher test of exact probability. There was a statistically higher prevalence of cerebral microhemorrhages in the group of boxers (3 of 42; 7.1%) than in nonboxing persons (0 of 37; 0%). This difference was not statistically significant, however (P = .2479; Fisher exact test). Although we detected more microhemorrhages in amateur boxers than in nonboxing persons, this difference did not prove to be significant.


c-Kit is essential for alveolar maintenance and protection from emphysema-like disease in mice.
Previously, we demonstrated a candidate region for susceptibility to airspace enlargement on mouse chromosome 5. However, the specific candidate genes within this region accounting for emphysema-like changes remain unrecognized. c-Kit is a receptor tyrosine kinase within this candidate gene region that has previously been recognized to contribute to the survival, proliferation, and differentiation of hematopoietic stem cells. Increases in the percentage of cells expressing c-Kit have previously been associated with protection against injury-induced emphysema. Determine whether genetic variants of c-Kit are associated with spontaneous airspace enlargement. Perform single-nucleotide polymorphism association studies in the mouse strains at the extremes of airspace enlargement phenotype for variants in c-Kit tyrosine kinase. Characterize mice bearing functional variants of c-Kit compared with wild-type controls for the development of spontaneous airspace enlargement. Epithelial cell proliferation was measured in culture. Upstream regulatory single-nucleotide polymorphisms in the divergent mouse strains were associated with the lung compliance difference observed between the extreme strains. c-Kit mutant mice (Kit(W-sh)/(W-sh)), when compared with genetic controls, developed altered lung histology, increased total lung capacity, increased residual volume, and increased lung compliance that persist into adulthood. c-Kit inhibition with imatinib attenuated in vitro proliferation of cells expressing epithelial cell adhesion molecule. Our findings indicate that c-Kit sustains and/or maintains normal alveolar architecture in the lungs of mice. In vitro data suggest that c-Kit can regulate epithelial cell clonal expansion. The precise mechanisms that c-Kit contributes to the development of airspace enlargement and increased lung compliance remain unclear and warrants further investigation.


Optimization and Application of a Biotinylation Method for Quantification of Plasma Membrane Expression of Transporters in Cells.
Quantitative proteomics, using LC-MS/MS, is increasingly used to quantify drug transporters present in tissues and cells. Most of these investigations quantify total transporter expression in the cells by utilizing a total membrane fraction, not only the plasma membrane. Here, we report development and optimization of a biotinylation method to quantify protein expression of transporters in the plasma membrane of cells. The Pierce cell surface isolation protocol was optimized for plasma membrane isolation. Incubation of OATP1B1-expressing CHO cells with 0.78 mg/mL of membrane impermeable biotinylation reagent (sulfo-NHS-SS-biotin) at 37°C for 1 h resulted in optimum isolation of the plasma membrane. Subsequently, the expression of transporters in the plasma membrane as a percent of the total was determined by quantitative proteomics using LC-MS/MS. Mean (±SD) plasma membrane expression of OATP1B1 in plated OATP1B1-expressing CHO, MDCKII, and HEK293 cells was found to be 79.7% (±4.7%), 67.7% (±12.2%), and 65.3% (±6.8%) of total cell OATP1B1 expression. Mean (±SD) plasma membrane expression of OATP1B3 in plated OATP1B3-expressing HEK293 cells, OATP2B1 in plated OATP2B1-expressing MDCKII cells, and sodium/taurocholate co-transporting polypeptide (NTCP) in plated NTCP-expressing CHO cells was 63.2% (±1.6%), 37.1% (±15.7%), and 71.7% (±1.2%), respectively. This method of quantifying transporter protein expression in the plasma membrane will be useful in the future to predict transporter-mediated drug disposition.


Megalin contributes to kidney accumulation and nephrotoxicity of colistin.
Interest has recently been shown again in colistin because of the increased prevalence of infections caused by multidrug-resistant Gram-negative bacteria. Although the potential for nephrotoxicity is a major dose-limiting factor in colistin use, little is known about the mechanisms that underlie colistin-induced nephrotoxicity. In this study, we focused on an endocytosis receptor, megalin, that is expressed in renal proximal tubules, with the aim of clarifying the role of megalin in the kidney accumulation and nephrotoxicity of colistin. We examined the binding of colistin to megalin by using a vesicle assay. The kidney accumulation, urinary excretion, and concentrations in plasma of colistin in megalin-shedding rats were also evaluated. Furthermore, we examined the effect of megalin ligands and a microtubule-depolymerizing agent on colistin-induced nephrotoxicity. We found that cytochrome c, a typical megalin ligand, inhibited the binding of colistin to megalin competitively. In megalin-shedding rats, renal proximal tubule colistin accumulation was decreased (13.5 ± 1.6 and 21.3 ± 2.6 μg in megalin-shedding and control rats, respectively). Coadministration of colistin and cytochrome c or albumin fragments resulted in a significant decrease in urinary N-acetyl-β-d-glucosaminidase (NAG) excretion, a marker of renal tubular damage (717.1 ± 183.9 mU/day for colistin alone, 500.8 ± 102.4 mU/day for cytochrome c with colistin, and 406.7 ± 156.7 mU/day for albumin fragments with colistin). Moreover, coadministration of colistin and colchicine, a microtubule-depolymerizing agent, resulted in a significant decrease in urinary NAG excretion. In conclusion, our results indicate that colistin acts as a megalin ligand and that megalin plays a key role in the accumulation in the kidney and nephrotoxicity of colistin. Megalin ligands may be new targets for the prevention of colistin-induced nephrotoxicity.


Therapeutic applications of non-myeloablative hematopoietic stem cell transplantation in malignant disease.
Allogeneic hematopoietic stem cell transplantation (HSCT) currently provides the only chance of curative therapy for many patients with hematological malignancies. Owing to the excess morbidity and mortality observed in less robust patients, this approach has traditionally been limited to younger patients without significant medical co-morbidities. Given the age profiles of patients with candidate hematological malignancies, a majority of patients are therefore ineligible for myeloablative therapy. Over the past few years the ability to achieve hematopoietic stem cell engraftment using immunosuppressive but non-myeloablative conditioning regimens has made it possible to consider a much broader group of patients for allogeneic HSCT. The discovery that the hematopoietic graft itself can contribute to the eradication of malignant cells through a graft-versus-malignancy (GVM) effect, independent of the conditioning regimen, has led to investigation of reduced intensity conditioning regimens for a variety of malignant diseases. Over the past few years there has been a dramatic increase in the number of patients undergoing reduced intensity conditioning for both malignant and nonmalignant conditions. This review will focus only on the use of such therapy for malignant disease.


Left ventricular efficiency after ligation of patent ductus arteriosus for premature infants.
The purpose of this study was to evaluate the hemodynamic changes in left ventricular function before and after patent ductus arteriosus ligation in premature infants with regard to the energetic efficiency of left ventricular pumping. Thirty-five premature infants who underwent patent ductus arteriosus ligation were enrolled in this study. Left ventricular efficiency was evaluated at 4 points: within 24 hours before patent ductus arteriosus ligation, within 24 hours after patent ductus arteriosus ligation, between postoperative days 2 and 4, and on postoperative day 7. The indices of contractility (end-systolic elastance) and afterload (effective arterial elastance) were approximated on the basis of the systemic blood pressure and systolic or diastolic left ventricular volume. The ratio of stroke work and pressure-volume area, representing the ventricular efficiency, was estimated using the following theoretic formula: the ratio of stroke work and pressure-volume area = 1/(1 + 0.5 ventriculoarterial coupling). Left ventricular efficiency was transiently deteriorated within 24 hours after patent ductus arteriosus ligation because of the marked increase of the afterload and the slight increase of contraction, and then recovered to preoperation levels by 2 to 4 days after patent ductus arteriosus ligation. Analysis of indices representing the afterload, contractility, and energetic efficiency of the left ventricle may provide practical information for the management of premature infants during the postoperative period after patent ductus arteriosus ligation.


A Java speech implementation of the Mini Mental Status Exam.
The Folstein Mini Mental Status Exam (MMSE) is a simple, widely used, verbally administered test to assess cognitive function. The Java Speech Application Programming Interface (JSAPI) is a new, cross-platform interface for both speech recognition and speech synthesis in the Java environment. To evaluate the suitability of the JSAPI for interactive, patient interview applications, a JSAPI implementation of the MMSE was developed. The MMSE contains questions that vary in structure in order to assess different cognitive functions. This question variability provided an excellent test-bed to evaluate the strengths and weaknesses of JSAPI. The application is based on Java platform 2 and a JSAPI interface to the IBM ViaVoice recognition engine. Design and implementations issues are discussed. Preliminary usability studies demonstrate that an automated MMSE maybe a useful screening tool for cognitive disorders and changes.


Impulsivity in internet addiction: a comparison with pathological gambling.
Internet addiction has been considered to be associated with poor impulse control. The aim of this study is to compare the trait impulsivity of those suffering from Internet addiction with that of individuals suffering from pathological gambling. Twenty-seven patients diagnosed with Internet addiction (age: 24.78±4.37 years), 27 patients diagnosed with pathological gambling (age: 25.67±3.97 years), and 27 healthy controls (age: 25.33±2.79 years) were enrolled in this study. All patients were men seeking treatment. Trait impulsivity and the severity of the Internet addiction and pathological gambling were measured by the Barratt Impulsiveness Scale-11, the Young's Internet Addiction Test, and the South Oaks Gambling Screen, respectively. The Beck Depression Inventory and the Beck Anxiety Inventory were also administered to all subjects. Our results show that those suffering from Internet addiction showed increased levels of trait impulsivity which were comparable to those of patients diagnosed with pathological gambling. Additionally, the severity of Internet addiction was positively correlated with the level of trait impulsivity in patients with Internet addiction. These results state that Internet addiction can be conceptualized as an impulse control disorder and that trait impulsivity is a marker for vulnerability to Internet addiction.


Small cell carcinoma of the ovary presenting in a urine cytology specimen: a case report.
Small cell carcinoma (SMCC) is rarely diagnosed in urine specimens. Cytologically, this tumor is similar to pulmonary SMCC. However, clinicopathologic correlation may be required to differentiate between primary urinary bladder SMCC and metastatic SMCC from a remote primary or secondary bladder involvement by direct extension of the tumor from nearby organs (prostate, uterus, or ovary). A unique case of a rare pulmonary-type ovarian SMCC, the tumor cells of which were detected in a voided urine specimen, is described herein. A 79-year-old female presented to the urologic clinic with a history of metastatic SMCC of unknown primary with hematuria. The voided urine specimen examination revealed tumor cells cytomorphologically consistent with small cell neuroendocrine carcinoma. Following cytologic diagnosis, cystoscopic examination and bladder biopsy were performed. The histopathology revealed a widely invasive tumor with a morphology typical of SMCC. The overlying urothelium was unremarkable. By immunohistochemistry, tumor cells were found positive for neuroendocrine markers, EMA and WT-1. The morphologic and immunohistochemical features of the tumor were most consistent with urinary bladder involvement by pulmonary-type primary ovarian SMCC. It is justified to think that SMCC cell detection in urine specimens does not necessarily imply their origin from primary bladder malignancy. Performing additional studies may be prudent in order to exclude secondary involvement of the bladder in this tumor as the correct diagnosis has significant clinical implications.