Evaluation of conformity of preformed orthodontic archwires and dental arch form.
The alterations in the arch form during treatment are dictated by the dimensions of the archwires. This study aimed to determine the mean arch dimensions of a sample of Pakistani subjects and to evaluate the conformity of preformed archwires with mandibular arch form. The dental records of 1,500 adult subjects were evaluated. The mandibular casts of 42 subjects (males = females = 21) with balanced facial profile, Class I occlusion, ideal overjet and overbite were included. Brackets were bonded on all teeth. Arch dimensions were evaluated at canines, first premolars, second premolars, first and second molars, with digital vernier calipers. The arch widths at the level of aforementioned teeth were evaluated on the digitized archwires, using the mean arch depths of the subjects. In males, the archwires were found to be wider at canines and premolars, and significantly narrower at first (p< 0.001, 95% CI = 2.03 - 5.74) and second molars (p< 0.001, 95% CI = 2.29 - 7.73) as compared to the arch dimensions of the mandibular casts. In females, the archwires were significantly narrower at canines (p< 0.001, 95% CI = 1.4 - 3.97), and first (p= 0.02, 95% CI = 0.402 - 4.41) and second molars (p< 0.001, 95% CI = 1.76 - 6.13). No single commercially available archwires evaluated in the present study conformed to the arch dimensions of our subjects. Utilization of the currently available archwires may result in unwarranted modification of arch form, which may lead to unstable post-treatment teeth position.


Risks of pneumonia in patients with asthma taking inhaled corticosteroids.
Inhaled corticosteroids (ICS) are the mainstay of asthma treatment. Studies in chronic obstructive pulmonary disease reported increased rates of pneumonia with ICS. Concerns exist about an increased pneumonia risk in patients with asthma taking ICS. To evaluate the risks of pneumonia in patients with asthma taking ICS. A retrospective analysis evaluated studies of the ICS budesonide in asthma. The primary data set were all double-blind, placebo-controlled trials lasting at least 3 months, involving budesonide (26 trials, n = 9,067 for budesonide; n = 5,926 for the comparator) sponsored by AstraZeneca. A secondary data set evaluated all double-blind trials lasting at least 3 months but without placebo control (60 trials, n = 33,496 for budesonide, n = 2,773 for fluticasone propionate). Cox proportional hazards regression modeling was used to estimate the relative effect of ICS on pneumonia adverse events (AEs) or serious adverse events (SAEs). In the primary data set, the occurrence of pneumonia AEs was 0.5% (rate 10.0 events/1,000 patient-years [TPY]) for budesonide and 1.2% (19.3 per TPY) for placebo (hazard ratio, 0.52; 95% confidence interval, 0.36-0.76; P < 0.001); the occurrence of pneumonia SAEs was 0.15% (2.9 per TPY) for budesonide and 0.13% (2.1 per TPY) for placebo (hazard ratio, 1.29; 95% confidence interval, 0.53-3.12; P = 0.58). In the secondary data set, the percentage of patients reporting pneumonia AEs was 0.70% (12.7 per TPY), whereas the percentage of patients reporting pneumonia SAEs was 0.17% (3.1 per TPY). There was no increased risk with higher budesonide doses or any difference between budesonide and fluticasone. There is no increased risk of pneumonia in patients with asthma, identified as an AE or SAE, in clinical trials using budesonide.


Promoting walking to school: results of a quasi-experimental trial.
To assess the impact of a combined intervention on children's travel behaviour, stage of behavioural change and motivations for and barriers to actively commuting to school. A quasi-experimental trial involving pre- and post-intervention mapping of routes to school by active and inactive mode of travel and surveys of "stage of behaviour change" and motivations for and barriers to actively commuting to school. The intervention school participated in a school-based active travel project for one school term. Active travel was integrated into the curriculum and participants used interactive travel-planning resources at home. The control school participated in before and after measurements but did not receive the intervention. Two primary schools in Scotland with similar socioeconomic and demographic profiles. Two classes of primary 5 children and their families and teachers. Post intervention, the mean distance travelled to school by walking by intervention children increased significantly from baseline, from 198 to 772 m (389% increase). In the control group mean distance walked increased from 242 to 285 m (17% increase). The difference between the schools was significant (t (38) = -4.679, p<0.001 (95% confidence interval -315 to -795 m)). Post intervention, the mean distance travelled to school by car by intervention children reduced significantly from baseline, from 2018 to 933 m (57.5% reduction). The mean distance travelled to school by car by control children increased from baseline, from 933 to 947 m (1.5% increase). The difference in the change between schools was significant (t (32) = 4.282, p<0.001 (95% confidence interval 445 to 1255 m)). Intervention was effective in achieving an increase in the mean distance travelled by active mode and a reduction in the mean distance travelled by inactive mode on school journey.


Metazoan parasite species in cultured mussel Mytilus galloprovincialis in the Thermaikos Gulf (North Aegean Sea, Greece).
This is the first study on parasites of cultured Mytilus galloprovincialis L. in Greek waters, and is based on samples collected every 2 to 3 mo between September 2000 and November 2001 at 3 stations in the Thermaikos Gulf. Each sample comprised 40 mussels. We found 4 metazoan parasite species: hydroid Eugymnanthea inquilina, gill turbellarian Urastoma cyprinae, trematode Proctoces maculatus and gut copepod Mytilicola intestinalis. Of 840 mussels examined, 406 (48.3%) mussels were harbouring hydroids of E. inquilina, 278 (33.1%) were infested with U. cyprinae, 94 (11.2%) were infested with M. intestinalis, and only 7 (0.8%) were infested with P. maculatus. The prevalence and intensity of these parasites were related to temperature and pollution. Mussels infested with these parasites had significantly lower condition indices than non-infested mussels; larger mussels were more often infested than smaller ones.


Conservative management of Morel-Lavallée lesion: a case study.
To illustrate the conservative clinical management of Morel-Lavallée lesion. A 34 year-old male presented with a forced flexion induced right knee injury that was diagnosed as a Morel-Lavallée lesion by way of Magnetic Resonance Imaging (MRI). The patient was co-managed by a sports medicine physician and a chiropractor with a combination of percutaneous aspiration, dry needling, soft tissue therapy, extracorporeal shockwave therapy (ESWT), and corrective exercise. The patient experienced improved function throughout chiropractic care. Almost complete resolution was achieved at 18 weeks after injury and maintained at 35 weeks after injury. Further research is needed to understand if this improvement in function is better than the natural history of this condition.


[A combined consecutive therapy with fosfomycin and sulbactam/cefoperazone for bacterial infections associated with hematological diseases].
A combination antibacterial therapy with fosfomycin (FOM) and sulbactam/cefoperazone (SBT/CPZ) was applied to 78 patients with severe infections associated with hematological diseases. In this protocol, FOM was followed by SBT/CPZ and each drug was administered for 1 hour intravenously and consecutively. Among 72 evaluable patients, 43 patients had acute leukemia, myeloblastic or lymphoblastic, 22 had malignant lymphoma, 3 had multiple myeloma, and 4 had other hematological diseases as underlying diseases. Bacterial infections diagnosed were sepsis in 21 patients, suspected sepsis in 47, and other infections in 4. The overall efficacy rate of this treatment was 72.2%, and those for individual infections were 66.7% for sepsis, 74.5% for suspected sepsis, and 75.0% for other infectious diseases. Among 22 bacteria separated from patients with sepsis, 78.6% (11/14 strains) were eradicated by this treatment. This protocol was also effective in 57.1% (8/14) of patients whose granulocyte count was less than 100/mm3 during the course of treatment as well as in 83.3% (15/18) of patients with granulocyte count over 500/mm3. There was no difference in effectiveness between those patients to whom G-CSF was administered and those to whom it was not (17/24, 70.8% vs 35/48, 72.9%). As an adverse reaction, a transient increase of GOT and/or GPT was observed in 2 patients (2.8%). The consecutive administration treatment of FOM and SBT/CPZ is thus an effective and safe regimen for the treatment of patients with hematological diseases complicated by severe infections.


Routes of Hendra Virus Excretion in Naturally-Infected Flying-Foxes: Implications for Viral Transmission and Spillover Risk.
Pteropid bats or flying-foxes (Chiroptera: Pteropodidae) are the natural host of Hendra virus (HeV) which sporadically causes fatal disease in horses and humans in eastern Australia. While there is strong evidence that urine is an important infectious medium that likely drives bat to bat transmission and bat to horse transmission, there is uncertainty about the relative importance of alternative routes of excretion such as nasal and oral secretions, and faeces. Identifying the potential routes of HeV excretion in flying-foxes is important to effectively mitigate equine exposure risk at the bat-horse interface, and in determining transmission rates in host-pathogen models. The aim of this study was to identify the major routes of HeV excretion in naturally infected flying-foxes, and secondarily, to identify between-species variation in excretion prevalence. A total of 2840 flying-foxes from three of the four Australian mainland species (Pteropus alecto, P. poliocephalus and P. scapulatus) were captured and sampled at multiple roost locations in the eastern states of Queensland and New South Wales between 2012 and 2014. A range of biological samples (urine and serum, and urogenital, nasal, oral and rectal swabs) were collected from anaesthetized bats, and tested for HeV RNA using a qRT-PCR assay targeting the M gene. Forty-two P. alecto (n = 1410) had HeV RNA detected in at least one sample, and yielded a total of 78 positive samples, at an overall detection rate of 1.76% across all samples tested in this species (78/4436). The rate of detection, and the amount of viral RNA, was highest in urine samples (>serum, packed haemocytes >faecal >nasal >oral), identifying urine as the most plausible source of infection for flying-foxes and for horses. Detection in a urine sample was more efficient than detection in urogenital swabs, identifying the former as the preferred diagnostic sample. The detection of HeV RNA in serum is consistent with haematogenous spread, and with hypothesised latency and recrudesence in flying-foxes. There were no detections in P. poliocephalus (n = 1168 animals; n = 2958 samples) or P. scapulatus (n = 262 animals; n = 985 samples), suggesting (consistent with other recent studies) that these species are epidemiologically less important than P. alecto in HeV infection dynamics. The study is unprecedented in terms of the individual animal approach, the large sample size, and the use of a molecular assay to directly determine infection status. These features provide a high level of confidence in the veracity of our findings, and a sound basis from which to more precisely target equine risk mitigation strategies.


Beta-cell dysfunction induced by chronic hyperglycemia. Current ideas on mechanism of impaired glucose-induced insulin secretion.
Non-insulin-dependent diabetes mellitus is characterized by abnormal beta-cell function. The characteristic secretory defect is a selective loss of glucose-induced insulin secretion. Substantial data have been generated in animal models to support the concept that chronic hyperglycemia causes the loss of glucorecognition (the so-called glucose toxicity hypothesis). This review summarizes the data supporting the concept of hyperglycemia-induced beta-cell dysfunction and then focuses on the ideas for the mechanism of the glucose unresponsiveness. The lack of access to islet tissue in humans means that these studies have all been conducted in animal models. Another major stumbling block continues to be the lack of in vitro systems that faithfully reproduce the secretory abnormalities that occur in vivo. Despite these limitations, many hypotheses are being investigated that span most of the major intracellular steps for glucose-induced insulin secretion, including abnormalities in glucose transport, storage, metabolism/oxidation, and the second messengers. No single hypothesis stands out as being able to explain all of the characteristics of the secretory abnormalities. In the last few years major advances have occurred in our knowledge about the events that normally cause glucose-induced insulin secretion. Similarly, biochemical and molecular tools have become available to probe the different steps. As better in vitro models of the selective glucose unresponsiveness become available, rapid progress can be expected in unraveling the biochemical basis for the loss of glucose responsiveness in diabetic rat models. The long-term hope is that this information will lead to innovative new strategies for the therapy of non-insulin-dependent diabetes mellitus.


CARI III inhibits tumor growth in a melanoma-bearing mouse model through induction of G0/G1 cell cycle arrest.
Mushroom-derived natural products have been used to prevent or treat cancer for millennia. In this study, we evaluated the anticancer effects of CARI (Cell Activation Research Institute) III, which consists of a blend of mushroom mycelia from Phellinus linteus grown on germinated brown rice, Inonotus obliquus grown on germinated brown rice, Antrodia camphorata grown on germinated brown rice and Ganoderma lucidum. Here, we showed that CARI III exerted anti-cancer activity, which is comparable to Dox against melanoma in vivo. B16F10 cells were intraperitoneally injected into C57BL6 mice to develop solid intra-abdominal tumors. Three hundred milligrams of the CARI III/kg/day p.o. regimen reduced tumor weight, comparable to the doxorubicin (Dox)-treated group. An increase in life span (ILS% = 50.88%) was observed in the CARI III-administered group, compared to the tumor control group. CARI III demonstrates anti-proliferative activity against B16F10 melanoma cells through inducing G0/G1 cell cycle arrest. CARI III inhibits the expression of cyclin D1, CDK4 and CDK2 and induces p21. Therefore, CARI III could be a potential chemopreventive supplement to melanoma patients.


Intratumoral hemorrhage and fibrosis in vestibular schwannoma: a possible mechanism for hearing loss.
Vestibular schwannomas (VSs) are benign lesions with an unpredictable natural history. Perhaps the greatest barrier to predicting which patients need treatment is our poor understanding of how these tumors cause hearing loss in the first place. In this case-control study, the authors investigated the relationship between preoperative hearing loss and histological changes such as intratumoral microhemorrhage and extensive fibrosis. From a prospectively collected database, the authors selected all patients with VS who had undergone microsurgical resection as their initial treatment for histopathologically confirmed VS. Histological specimens obtained in 274 of these patients were systematically reviewed by a blinded neuropathologist who graded the extent of microhemorrhage and fibrosis in these tumors. The effect of these variables on preoperative hearing loss was studied using binary logistic regression. On univariate analysis, patients with extensive intratumoral microhemorrhage or fibrosis (p < 0.0001), patients with larger tumors (p < 0.05), and patients 65 years of age or older (p < 0.05) were significantly more likely to have unserviceable hearing at the time of surgery. On multivariate analysis, only patients with extensive intratumoral microhemorrhage or fibrosis had an increased risk of having unserviceable hearing at the time of surgery (OR 3.72, 95% CI 1.3-10; p = 0.01). Older age and tumor size greater than 3 cm were not statistically significant risk factors for hearing loss, controlling for the effect of microhemorrhage and fibrosis. In this study, the authors have demonstrated a correlation between the extent of nonneoplastic histological changes, such as microhemorrhage and fibrosis, and hearing loss. This alternate hypothesis has the potential to explain many of the exceptions to previously described mechanisms of hearing loss in patients with VS. The advent of high-resolution MR imaging technology to identify microhemorrhages may provide a method to screen for patients with VS at risk for hearing loss.


Second-harmonic generation from magnetic metamaterials.
We observe second-harmonic generation from metamaterials composed of split-ring resonators excited at 1.5-micrometer wavelength. Much larger signals are detected when magnetic-dipole resonances are excited, as compared with purely electric-dipole resonances. The experiments are consistent with calculations based on the magnetic component of the Lorentz force exerted on metal electrons-an intrinsic second-harmonic generation mechanism that plays no role in natural materials. This unusual mechanism becomes relevant in our work as a result of the enhancement and the orientation of the local magnetic fields associated with the magnetic-dipole resonances of the split-ring resonators.


Endocrine and radiological studies in patients with molecularly confirmed CHARGE syndrome.
CHARGE syndrome is a complex of congenital malformations, and CHD7 has been reported as a major gene involved in the etiology. We performed endocrine and radiological studies to determine whether endocrinological disorders such as hypogonadotropic hypogonadism, GH deficiency, or hypothyroidism are involved and also whether olfactory bulb hypoplasia and semicircular canal aplasia are major signs in patients with molecularly confirmed CHARGE syndrome. Clinical features, endocrinological assessments, and radiological abnormalities in eight children (five boys and three girls) whose molecular analyses were available were evaluated among 15 children clinically diagnosed with CHARGE syndrome at our institute. We identified heterozygous CHD7 mutations in all patients screened for mutations. Four boys had micropenis and/or cryptorchidism. One was diagnosed with GH deficiency, and the other was diagnosed with hypothyroidism. Computed tomography findings revealed aplasia of the semicircular canals. Magnetic resonance imaging studies of the olfactory bulb region revealed abnormal olfactory sulci and bulb development in all children. We suggest that hypogonadism, GH deficiency, and hypothyroidism could be possible endocrinological defects in patients with CHD7 mutations and that olfactory bulb hypoplasia as well as semicircular canal aplasia should be considered as a major sign for CHARGE syndrome and recommend a computed tomography scan of the temporal bone and magnetic resonance imaging study of the olfactory bulb region.


Local TNF causes NFATc1-dependent cholesterol-mediated podocyte injury.
High levels of circulating TNF and its receptors, TNFR1 and TNFR2, predict the progression of diabetic kidney disease (DKD), but their contribution to organ damage in DKD remains largely unknown. Here, we investigated the function of local and systemic TNF in podocyte injury. We cultured human podocytes with sera collected from DKD patients, who displayed elevated TNF levels, and focal segmental glomerulosclerosis (FSGS) patients, whose TNF levels resembled those of healthy patients. Exogenous TNF administration or local TNF expression was equally sufficient to cause free cholesterol-dependent apoptosis in podocytes by acting through a dual mechanism that required a reduction in ATP-binding cassette transporter A1-mediated (ABCA1-mediated) cholesterol efflux and reduced cholesterol esterification by sterol-O-acyltransferase 1 (SOAT1). TNF-induced albuminuria was aggravated in mice with podocyte-specific ABCA1 deficiency and was partially prevented by cholesterol depletion with cyclodextrin. TNF-stimulated free cholesterol-dependent apoptosis in podocytes was mediated by nuclear factor of activated T cells 1 (NFATc1). ABCA1 overexpression or cholesterol depletion was sufficient to reduce albuminuria in mice with podocyte-specific NFATc1 activation. Our data implicate an NFATc1/ABCA1-dependent mechanism in which local TNF is sufficient to cause free cholesterol-dependent podocyte injury irrespective of TNF, TNFR1, or TNFR2 serum levels.


Evidence of gene-gene interaction and age-at-diagnosis effects in type 1 diabetes.
The common genetic loci that independently influence the risk of type 1 diabetes have largely been determined. Their interactions with age-at-diagnosis of type 1 diabetes, sex, or the major susceptibility locus, HLA class II, remain mostly unexplored. A large collection of more than 14,866 type 1 diabetes samples (6,750 British diabetic individuals and 8,116 affected family samples of European descent) were genotyped at 38 confirmed type 1 diabetes-associated non-HLA regions and used to test for interaction of association with age-at-diagnosis, sex, and HLA class II genotypes using regression models. The alleles that confer susceptibility to type 1 diabetes at interleukin-2 (IL-2), IL2/4q27 (rs2069763) and renalase, FAD-dependent amine oxidase (RNLS)/10q23.31 (rs10509540), were associated with a lower age-at-diagnosis (P = 4.6 × 10⁻⁶ and 2.5 × 10⁻⁵, respectively). For both loci, individuals carrying the susceptible homozygous genotype were, on average, 7.2 months younger at diagnosis than those carrying the protective homozygous genotypes. In addition to protein tyrosine phosphatase nonreceptor type 22 (PTPN22), evidence of statistical interaction between HLA class II genotypes and rs3087243 at cytotoxic T-lymphocyte antigen 4 (CTLA4)/2q33.2 was obtained (P = 7.90 × 10⁻⁵). No evidence of differential risk by sex was obtained at any loci (P ≥ 0.01). Statistical interaction effects can be detected in type 1 diabetes although they provide a relatively small contribution to our understanding of the familial clustering of the disease.


Role of molecular testing in the multidisciplinary diagnostic approach of ichthyosis.
The term ichthyosis describes a generalized disorder of cornification characterized by scaling and/or hyperkeratosis of different skin regions. Mutations in a broad group of genes related to keratinocyte differentiation and epidermal barrier function have been demonstrated to play a causative role in disease development. Ichthyosis may be classified in syndromic or non-syndromic forms based on the occurrence or absence of extracutaneous signs. In this setting, the diagnosis of ichthyosis is an integrated multistep process requiring a multidisciplinary approach in order to formulate the appropriate diagnostic hypothesis and to address the genetic testing. Due to the complex features of the different ichthyoses and the high number of genes involved we have investigated a group of 64 patients, affected by syndromic and non-syndromic diseases, using Next Generation Sequencing as a new tool for the molecular diagnosis. Using this innovative molecular approach we were able to find pathogenic mutations in 53 out of 64 patients resulting in 82.8 % total detection rate. An interesting result from the analysis of the data is the high rate of novel sequence variations found compared to known mutations and the relevant rate of homozygous mutations. The possibility to analyze a large number of genes associated with various diseases allows to study cases with phenotypes not well-determined, giving the opportunity to make new genotype-phenotype correlation. In some cases there were discrepancies between clinical features and histology or electron microscopy and only molecular analysis allowed to definitively resolve the diagnostic dilemma. The genetic diagnosis of ichthyosis leads to a more accurate and effective genetic counseling, allowing correct evaluation of the risk of recurrence, particularly in families with consanguineous background.


Passive mechanical forces control cell-shape change during Drosophila ventral furrow formation.
During Drosophila gastrulation, the ventral mesodermal cells constrict their apices, undergo a series of coordinated cell-shape changes to form a ventral furrow (VF) and are subsequently internalized. Although it has been well documented that apical constriction is necessary for VF formation, the mechanism by which apical constriction transmits forces throughout the bulk tissue of the cell remains poorly understood. In this work, we develop a computational vertex model to investigate the role of the passive mechanical properties of the cellular blastoderm during gastrulation. We introduce to our knowledge novel data that confirm that the volume of apically constricting cells is conserved throughout the entire course of invagination. We show that maintenance of this constant volume is sufficient to generate invagination as a passive response to apical constriction when it is combined with region-specific elasticities in the membranes surrounding individual cells. We find that the specific sequence of cell-shape changes during VF formation is critically controlled by the stiffness of the lateral and basal membrane surfaces. In particular, our model demonstrates that a transition in basal rigidity is sufficient to drive VF formation along the same sequence of cell-shape change that we observed in the actual embryo, with no active force generation required other than apical constriction.


Chameleon-like elastomers with molecularly encoded strain-adaptive stiffening and coloration.
Active camouflage is widely recognized as a soft-tissue feature, and yet the ability to integrate adaptive coloration and tissuelike mechanical properties into synthetic materials remains elusive. We provide a solution to this problem by uniting these functions in moldable elastomers through the self-assembly of linear-bottlebrush-linear triblock copolymers. Microphase separation of the architecturally distinct blocks results in physically cross-linked networks that display vibrant color, extreme softness, and intense strain stiffening on par with that of skin tissue. Each of these functional properties is regulated by the structure of one macromolecule, without the need for chemical cross-linking or additives. These materials remain stable under conditions characteristic of internal bodily environments and under ambient conditions, neither swelling in bodily fluids nor drying when exposed to air.


Level of agreement between physical activity levels measured by ActiHeart and the International Physical Activity Questionnaire in persons with intellectual disability.
The purpose of this study was to determine the level of agreement between objective physical activity (PA) (ActiHeart®) and subjective proxy-respondent International Physical Activity Questionnaire-short version (IPAQ-S) data in adults with intellectual disabilities (IDs). Fifty-eight participants wore ActiHeart® monitors for seven consecutive days. Caregivers of each participant completed the IPAQ-S on behalf of the participant. Total PA, time spent in light, moderate, and vigorous activity as well as time spent being sedentary were assessed by the IPAQ-S and the ActiHeart®. Results were compared by means of correlation analyses. The level of agreement was presented with Bland-Altman plots. Objective PA (ActiHeart®) was higher (225.57 ± 91.96 min/week) than IPAQ-S PA reported by care-givers (177.06 ± 309.17 min/week). Weak significant correlations were observed between the ActiHeart® and IPAQ-S instruments for sedentary behavior (r = 0.31; p = 0.04); no significant correlations for light (r= -0.04; p = 0.8), moderate (r= -0.07; p = 0.63), or vigorous PA (r= -0.2; p = 0.18) were found. Limited agreement between objectively determine PA (ActiHeart®) and IPAQ-S was found. IPAQ-S is inaccurate when determining PA in persons with ID as it significantly underestimates the true levels of PA in this cohort. Implications for Rehabilitation Persons with intellectual disability (ID) report insufficient physical activity for health benefits. Physical activity is often determined by means of subjective proxy reporting. Objective physical activity measurements by means of combined heart rate and accelerometer are necessary to determine accurate levels of physical activity in persons with ID. Exercise interventions should be based on objective physical activity measurements.


Methods for Genome-Wide Methylome Profiling of Campylobacter jejuni.
Methylation has a profound role in the regulation of numerous biological processes in bacteria including virulence. The study of methylation in bacteria has greatly advanced thanks to next-generation sequencing technologies. These technologies have expedited the process of uncovering unique features of many bacterial methylomes such as characterizing previously uncharacterized methyltransferases, cataloging genome-wide DNA methylations in bacteria, identifying the frequency of methylation at particular genomic loci, and revealing regulatory roles of methylation in the biology of various bacterial species. For instance, methylation has been cited as a potential source for the pathogenicity differences observed in C. jejuni strains with syntenic genomes as seen in recent publications. Here, we describe the methodology for the use of Pacific Biosciences' single molecule real-time (SMRT) sequencing for detecting methylation patterns in C. jejuni and bioinformatics tools to profile its methylome.


Side population cells isolated from mesenchymal neoplasms have tumor initiating potential.
Although many cancers are maintained by tumor-initiating cells, this has not been shown for mesenchymal tumors, in part due to the lack of unique surface markers that identify mesenchymal progenitors. An alternative technique to isolate stem-like cells is to isolate side population (SP) cells based on efflux of Hoechst 33342 dye. We examined 29 mesenchymal tumors ranging from benign to high-grade sarcomas and identified SP cells in all but six samples. There was a positive correlation between the percentage of SP cells and the grade of the tumor. SP cells preferentially formed tumors when grafted into immunodeficient mice, and only cells from tumors that developed from the SP cells had the ability to initiate tumor formation upon serial transplantation. Although SP cells are able to efflux rhodamine dye in addition to Hoechst 33342, we found that the ability to efflux rhodamine dye did not identify a population of cells enriched for tumor-initiating capacity. Here, we identify a subpopulation of cells within a broad range of benign and malignant mesenchymal tumors with tumor-initiating capacity. In addition, our data suggest that the proportion of SP cells could be used as a prognostic factor and that therapeutically targeting this subpopulation of cells could be used to improve patient outcome.


3D-printed IFN-γ-loading calcium silicate-β-tricalcium phosphate scaffold sequentially activates M1 and M2 polarization of macrophages to promote vascularization of tissue engineering bone.
To promote vascularization of tissue-engineered bone, IFN-γ polarizing macrophages to M1 was loaded on 5% calcium silicate/β-tricalcium phosphate (CaSiO3-β-TCP) scaffolds. IFN-γ and Si released from the scaffold were designed to polarize M1 and M2 macrophages, respectively. β-TCP, CaSiO3-β-TCP, and IFN-γ@CaSiO3-β-TCP were fabricated and biocompatibilities were evaluated. Polarizations of macrophages were detected by flow cytometry. Human umbilical vein endothelial cells with GFP were cultured and induced on Matrigel with conditioned culture medium extracted from culture of macrophages loaded on scaffolds for evaluating angiogenesis. Four weeks after the scaffolds were subcutaneously implanted into C57B1/6, vascularization was evaluated by visual observation, hematoxylin and eosin staining, as well as immunohistochemistry of CD31. The results showed that IFN-γ@CaSiO3-β-TCP scaffolds released IFN-γ in the early stage (1-3 days) to stimulate macrophages to M1 polarization, followed by release of Si inducing macrophages to M2 polarization while scaffolds degraded. The activation of M1/M2 allows macrophages to secrete more cytokines, including VEGF, CXCL12 and PDGF-BB. The IFN-γ@CaSiO3-β-TCP scaffolds formed more blood vessels in vitro and in vivo compared to the control groups. The study indicated that the design of tissue-engineered scaffolds with immunomodulatory function utilized host macrophages to increase vascularization of tissue-engineered bone, providing a new strategy for accelerating vascularization and osteogenesis of tissue-engineered scaffolds and showing the potential for treatment of major bone defects. A 3-D printed immunomodulatory scaffold was designed for repair of massive bone defects. Through the release of interferon γ and silicon ions, the new immunomodulatory scaffold promoted the M1 and M2 polarization of macrophages, boosting angiogenesis. This scaffold provided a new strategy for accelerating vascularization and osteogenesis of tissue-engineered scaffolds and showing the potential for treatment of major bone defects.


Induction of hapten-specific tolerance by interleukin 10 in vivo.
Interleukin 10 (IL-10) is released during the induction phase of contact sensitivity and was shown in prior functional studies to convert epidermal Langerhans cells (LC) from potent inducers of primary immune responses to specifically tolerizing cells in vitro. To investigate whether IL-10 also subserves the function of a tolerizing agent in vivo ears of BALB/c or C3H mice were injected intradermally with 1-2 micrograms of recombinant mouse (rm)IL-10 8 h before epicutaneous application of 3% trinitrochlorobenzene (TNCB; a contact allergen). As a control, mice were injected with phosphate-buffered saline or IL-10 plus neutralizing amounts of anti-IL-10 mAb. 5 d later, mice were challenged with 1% TNCB on contralateral ears and ear swelling response was measured 24 h later. Whereas control-treated mice showed a normal ear swelling response to epicutaneous challenge (delta mm-2 = 25 +/- 5), ear swelling response of IL-10-treated animals was significantly inhibited (delta mm-2 = 3 +/- 2). Coinjection of IL-10-specific mAb together with rmIL-10 completely abrogated this effect. To differentiate between a state of nonresponsiveness and induction of tolerance by IL-10, mice initially treated with IL-10 and TNCB were resensitized with 3% TNCB in the absence of any treatment after 14 d of rest (group 1). Again mice were challenged 5 d later and ear swelling responses were tested. Whereas control mice treated with allergen alone (group 2) showed a good swelling response (delta mm-2 = 28 +/- 6), IL-10-treated mice (group 1) showed a minimal response towards application of allergen (delta mm-2 = 4 +/- 2). To show that anergy induction by IL-10 was antigen-specific, mice initially treated with IL-10 plus TNCB were exposed to 0.5% dinitrofluorobenzene (DNFB) 14 d later (group 1). After challenge with 0.1% DNFB, IL-10-treated mice showed an ear swelling response (delta mm-2 = 13 +/- 3; group 1) similar to that of control mice only sensitized with DNFB (delta mm-2 = 14 +/- 3; group 3). In an attempt to show the induction of antigen-specific tolerance in these mice in vitro, regional lymph nodes of mice initially treated with TNCB plus IL-10 (group 1) and control-treated mice (groups 2 and 3) were prepared and cultured in the presence of TNBS, dinitrobenzene sulfonate (DNBS), or medium to measure antigen-specific proliferation.(ABSTRACT TRUNCATED AT 400 WORDS)


Skill shortages in health: innovative solutions using vocational education and training.
This article reports findings of a project funded by the Australian National Council for Vocational Education Research. The project explores solutions to current and projected skills shortages within the health and community services sector, from a vocational education and training perspective. Its purpose is to locate, analyse and disseminate information about innovative models of health training and service delivery that have been developed in response to skill shortages. The article begins with a brief overview of Australian statistics and literature on the structure of the national health workforce and perceived skill shortages. The impact of location (state and rurality), demographics of the workforce, and other relevant factors, on health skill shortages is examined. Drawing on a synthesis of the Australian and international literature on innovative and effective models for addressing health skill shortages and nominations by key stakeholders within the health sector, over 70 models were identified. The models represent a mixture of innovative service delivery models and training solutions from Australia, as well as international examples that could be transposed to the Australian context. They include the skill ecosystem approach facilitated by the Australian National Training Authority Skill Ecosystem Project. Models were selected to represent diversity in terms of the nature of skill shortage addressed, barriers overcome in development of the model, healthcare specialisations, and different customer groups. Key barriers to the development of innovative solutions to skills shortages identified were: policy that is not sufficiently flexible to accommodate changing workplace needs; unwillingness to risk take in order to develop new models; delays in gaining endorsement/accreditation; current vocational education and training (VET) monitoring and reporting systems; issues related to working in partnership, including different cultures, ways of operating, priorities and timelines; workplace culture that is resistant to change; and organisational boundaries. For training-only models, additional barriers were: technology; low educational levels of trainees; lack of health professionals to provide training and/or supervision; and cost of training. Key enhancers for the development of models were identified as: commitment by all partners and co-location of partners; or effective communication channels. Key enhancers for model effectiveness were: first considering work tasks, competencies and job (re)design; high profile of the model within the community; community-based models; cultural fit; and evidence of direct link between skills development and employment, for example VET trained aged care workers upskilling for other health jobs. For training only models, additional enhancers were flexibility of partners in accommodating needs of trainees; low training costs; experienced clinical supervisors; and the provision of professional development to trainers. There needs to be a balance between short-term solutions to current skill shortages (training only), and medium to longer term solutions (job redesign, holistic approaches) that also address projected skills shortages. Models that focus on addressing skills shortages in aged care can provide a broad pathway to careers in health. Characteristics of models likely to be effective in addressing skill shortages are: responsibility for addressing skills shortage is shared between the health sector, education and training organisations and government, with employers taking a proactive role; the training component is complemented by a focus on retention of workers; models are either targeted at existing employees or identify a target group(s) who may not otherwise have considered a career in health.


Insulin redirects differentiation from cardiogenic mesoderm and endoderm to neuroectoderm in differentiating human embryonic stem cells.
Human embryonic stem cells (hESC) can proliferate indefinitely while retaining the capacity to form derivatives of all three germ layers. We have reported previously that hESC differentiate into cardiomyocytes when cocultured with a visceral endoderm-like cell line (END-2). Insulin/insulin-like growth factors and their intracellular downstream target protein kinase Akt are known to protect many cell types from apoptosis and to promote proliferation, including hESC-derived cardiomyocytes. Here, we show that in the absence of insulin, a threefold increase in the number of beating areas was observed in hESC/END-2 coculture. In agreement, the addition of insulin strongly inhibited cardiac differentiation, as evidenced by a significant reduction in beating areas, as well as in alpha-actinin and beta-myosin heavy chain (beta-MHC)-expressing cells. Real-time reverse transcription-polymerase chain reaction and Western blot analysis showed that insulin inhibited cardiomyogenesis in the early phase of coculture by suppressing the expression of endoderm (Foxa2, GATA-6), mesoderm (brachyury T), and cardiac mesoderm (Nkx2.5, GATA-4). In contrast to previous reports, insulin was not sufficient to maintain hESC in an undifferentiated state, since expression of the pluripotency markers Oct3/4 and nanog declined independently of the presence of insulin during coculture. Instead, insulin promoted the expression of neuroectodermal markers. Since insulin triggered sustained phosphorylation of Akt in hESC, we analyzed the effect of an Akt inhibitor during coculture. Indeed, the inhibition of Akt or insulin-like growth factor-1 receptor reversed the insulin-dependent effects. We conclude that in hESC/END-2 cocultures, insulin does not prevent differentiation but favors the neuroectodermal lineage at the expense of mesendodermal lineages.


The influence of conjugation variables on the design and immunogenicity of a glycoconjugate vaccine against Salmonella Typhi.
In recent years there have been major efforts to develop glycoconjugate vaccines based on the Vi polysaccharide that will protect against Salmonella enterica Typhi infections, particularly typhoid fever, which remains a major public health concern in low-income countries. The design of glycoconjugate vaccines influences the immune responses they elicit. Here we systematically test the response in mice to Vi glycoconjugates that differ in Vi chain length (full-length and fragmented), carrier protein, conjugation chemistry, saccharide to protein ratio and size. We show that the length of Vi chains, but not the ultimate size of the conjugate, has an impact on the anti-Vi IgG immune response induced. Full-length Vi conjugates, independent of the carrier protein, induce peak IgG responses rapidly after just one immunization, and secondary immunization does not enhance the magnitude of these responses. Fragmented Vi linked to CRM197 and diphtheria toxoid, but not to tetanus toxoid, gives lower anti-Vi antibody responses after the first immunization than full-length Vi conjugates, but antibody titres are similar to those induced by full-length Vi conjugates following a second dose. The chemistry to conjugate Vi to the carrier protein, the linker used, and the saccharide to protein ratio do not significantly alter the response. We conclude that Vi length and carrier protein are the variables that influence the anti-Vi IgG response to immunization the most, while other parameters are of lesser importance.


Cancer in migrant and seasonal hired farm workers.
Studies of cancer among farm workers are difficult to conduct and interpret given the unique nature of this occupational group. The transitory nature of the work, high levels of poverty, and lack of legal documentation make epidemiologic studies difficult to accomplish. Nevertheless, this workforce in the United States, which numbers as much as 3 million persons, is a high risk population due to exposures to numerous toxic substances, including excessive sunlight, heat, dangerous machinery, fumes, fertilizers, dust, and pesticides. We summarize characteristics of farm workers (i.e., demographics, health care) from the National Agricultural Workers Survey (NAWS) and the California Agricultural Workers Survey (CAWS) and present findings from a series of studies conducted among farm workers in California. The epidemiology literature was reviewed and methods for a unique farm worker union-based epidemiologic study are presented. Farm workers in California and the rest of the United States, many of whom are seasonal and migrant workers are at elevated risk for numerous forms of cancer compared to the general population and specific pesticides may be associated with this altered risk. Elevated risks have been found for lymphomas and prostate, brain, leukemia, cervix, and stomach cancers.


Drugs of abuse and Parkinson's disease.
The term "drug of abuse" is highly contextual. What constitutes a drug of abuse for one population of patients does not for another. It is therefore important to examine the needs of the patient population to properly assess the status of drugs of abuse. The focus of this article is on the bidirectional relationship between patients and drug abuse. In this paper we will introduce the dopaminergic systems of the brain in Parkinson's and the influence of antiparkinsonian drugs upon them before discussing this synergy of condition and medication as fertile ground for drug abuse. We will then examine the relationship between drugs of abuse and Parkinson's, both beneficial and deleterious. In summary we will draw the different strands together and speculate on the future merit of current drugs of abuse as treatments for Parkinson's disease.


A comparative study of macrobenthic community from harbours along the central west coast of India.
Harbours are heavily stressed coastal habitats characterised by high concentration of contaminant and low diversity of benthic community. The west coast of India harbours most of the major harbours compared to the east coast. Very few studies have compared the macrobenthic community between different Indian harbours. The present study was therefore conducted in three important harbour (Ratnagiri, Goa, Karwar) along the central west coast of India. The paper discusses the health status of the three harbours diagnosed using various biotic indices. Sediment samples were collected using van Veen grab (0.11 m(2)) on board CRV Sagar Sukti. A total of 55 macrobenthic taxa were identified and were numerically dominated by polychaete. Biomass was high (0.14-145.7 g m(-2)) and was made largely by echiurans (>80%). Overall, polychaete dominated the macrobenthic diversity. Opportunistic P.pinnata, Notomastus sp. and Mediomastus sp., dominated the macrobenthic community responding to the increased in the harbour. Biotic indices (Polychaete:Amphipod ratio, ABC curve and geometric class abundance) and the dominance of opportunistic species indicate that, the three harbours are under stress from anthropogenic activities.


Visualized absorption of anti-atherosclerotic dipeptide, Trp-His, in Sprague-Dawley rats by LC-MS and MALDI-MS imaging analyses.
The basic dipeptide, Trp-His, was found to show an in vivo anti-atherosclerotic effect when orally administered to apo E-deficient mice. In addition, this dipeptide causes vasorelaxation in contracted rat aorta via suppression of intracellular Ca(2+) signaling cascades. In this study, we attempted to determine whether Trp-His can be absorbed after single oral administration in Sprague-Dawley (SD) rats. Trp-His and His-Trp (10 or 50 mg/kg) was orally administered to 8-week-old male SD rats. Both peptides in plasma were assayed by LC-MS/MS in combination with 2,4,6-trinitrobenzene sulfonate derivatization technique. In vitro transport experiments using Caco-2 cell monolayers were performed to evaluate the apparent permeability (Papp ). A phytic acid-aided MALDI-MS imaging (MSI) was conducted to visualize the distribution of dipeptides in the rat intestinal membrane. Trp-His was absorbed intact into SD rat blood, showing a maximal level at 1 h after administration at 10 mg/kg dose (Cmax , 28.7 ± 8.9 pmol/mL-plasma; area under the curve, 71.3 ± 18.7 pmol·h/mL-plasma). In contrast, His-Trp was surprisingly not detected, although the Papp was compatible to that of Trp-His. MSI analysis provided crucial evidence that Trp-His was visualized in the overall intestinal membrane. The Trp-His peptide was not visualized in the presence of Gly-Sar, which is a model peptide that is transported via the intestinal proton-coupled peptide transporter 1 (PepT1) transporter. The His-Trp molecular ion was not observed at the intestinal membrane. The MSI analysis illustrated that there is no absorption of His-Trp due to its unexpected hydrolysis by brush border proteases. To the best of our knowledge, this is the first study demonstrating that the vasoactive Trp-His is preferably transported across the rat intestinal membrane by PepT1 and is absorbed intact into the circulation. However, no absorption of His-Trp, a reverse sequence of absorbable Trp-His, is observed owing to hydrolysis by intestinal proteases. This suggests that the bioavailability of peptides may be determined in part by their protease resistance in the intestinal membrane.


Pilot evaluation of a walking school bus program in a low-income, urban community.
To evaluate the impact of a walking school bus (WSB) program on student transport in a low-income, urban neighborhood. The design was a controlled, quasi-experimental trial with consecutive cross-sectional assessments. The setting was three urban, socioeconomically disadvantaged, public elementary schools (1 intervention vs. 2 controls) in Seattle, Washington, USA. Participants were ethnically diverse students in kindergarten-5th grade (aged 5-11 years). The intervention was a WSB program consisting of a part-time WSB coordinator and parent volunteers. Students' method of transportation to school was assessed by a classroom survey at baseline and one-year follow-up. The Pearson Chi-squared test compared students transported to school at the intervention versus control schools at each time point. Due to multiple testing, we calculated adjusted p-values using the Ryan-Holm stepdown Bonferroni procedure. McNemar's test was used to examine the change from baseline to 12-month follow-up for walking versus all other forms of school transport at the intervention or control schools. At baseline, the proportions of students (n = 653) walking to the intervention (20% +/- 2%) or control schools (15% +/- 2%) did not differ (p = 0.39). At 12-month follow up, higher proportions of students (n = 643, p = 0.001)) walked to the intervention (25% +/- 2%) versus the control schools (7% +/- 1%). No significant changes were noted in the proportion of students riding in a car or taking the school bus at baseline or 12-month follow up (all p > 0.05). Comparing baseline to 12-month follow up, the numbers of students who walked to the intervention school increased while the numbers of students who used the other forms of transport did not change (p < 0.0001). In contrast, the numbers of students who walked to the control schools decreased while the numbers of students who used the other forms of transport did not change (p < 0.0001). A WSB program is a promising intervention among urban, low-income elementary school students that may promote favorable changes toward active transport to school. ClinicalTrials.gov NCT00402701.


The antimicrobial activity of prototype modified honeys that generate reactive oxygen species (ROS) hydrogen peroxide.
Antimicrobial resistance continues to be a global issue in healthcare organisations. Honey has long been shown to possess wound healing and antimicrobial properties that are dependent on a number of physical and chemical properties of the honey. We tested the antimicrobial activity of a medicinal honey, Surgihoney® (SH) and two prototype modified honeys made by Apis mellifera (honeybee) against Staphylococcus aureus (NCIMB 9518). We also examined the modified honey prototypes for the ability to generate reactive oxygen species (ROS) by changing the level of production of hydrogen peroxide from the samples. Surgihoney® (SH) was compared with two modified honeys, Prototype 1 (PT1) and Prototype 2 (PT2) using a bioassay method against a standard strain of Staphylococcus aureus. Further work studied the rate of generation of ROS hydrogen peroxide from these preparations. Surgihoney® antimicrobial activity was shown to be largely due to ROS hydrogen peroxide production. By modification of Surgihoney®, two more potent honey prototypes were shown to generate between a two- and three-fold greater antibacterial activity and up to ten times greater ROS peroxide activity. Surgihoney® is a clinically available wound antiseptic dressing that shows good antimicrobial activity. Two further honey prototypes have been shown to have antimicrobial activity that is possible to be enhanced due to demonstrated increases in ROS peroxide activity.


Colorectal cancer liver metastatic growth depends on PAD4-driven citrullination of the extracellular matrix.
Citrullination of proteins, a post-translational conversion of arginine residues to citrulline, is recognized in rheumatoid arthritis, but largely undocumented in cancer. Here we show that citrullination of the extracellular matrix by cancer cell derived peptidylarginine deiminase 4 (PAD4) is essential for the growth of liver metastases from colorectal cancer (CRC). Using proteomics, we demonstrate that liver metastases exhibit higher levels of citrullination and PAD4 than unaffected liver, primary CRC or adjacent colonic mucosa. Functional significance for citrullination in metastatic growth is evident in murine models where inhibition of citrullination substantially reduces liver metastatic burden. Additionally, citrullination of a key matrix component collagen type I promotes greater adhesion and decreased migration of CRC cells along with increased expression of characteristic epithelial markers, suggesting a role for citrullination in promoting mesenchymal-to-epithelial transition and liver metastasis. Overall, our study reveals the potential for PAD4-dependant citrullination to drive the progression of CRC liver metastasis.


Prion seeding activities of mouse scrapie strains with divergent PrPSc protease sensitivities and amyloid plaque content using RT-QuIC and eQuIC.
Different transmissible spongiform encephalopathy (TSE)-associated forms of prion protein (e.g. PrP(Sc)) can vary markedly in ultrastructure and biochemical characteristics, but each is propagated in the host. PrP(Sc) propagation involves conversion from its normal isoform, PrP(C), by a seeded or templated polymerization mechanism. Such a mechanism is also the basis of the RT-QuIC and eQuIC prion assays which use recombinant PrP (rPrP(Sen)) as a substrate. These ultrasensitive detection assays have been developed for TSE prions of several host species and sample tissues, but not for murine models which are central to TSE pathogenesis research. Here we have adapted RT-QuIC and eQuIC to various murine prions and evaluated how seeding activity depends on glycophosphatidylinositol (GPI) anchoring and the abundance of amyloid plaques and protease-resistant PrP(Sc) (PrP(Res)). Scrapie brain dilutions up to 10(-8) and 10(-13) were detected by RT-QuIC and eQuIC, respectively. Comparisons of scrapie-affected wild-type mice and transgenic mice expressing GPI anchorless PrP showed that, although similar concentrations of seeding activity accumulated in brain, the heavily amyloid-laden anchorless mouse tissue seeded more rapid reactions. Next we compared seeding activities in the brains of mice with similar infectivity titers, but widely divergent PrP(Res) levels. For this purpose we compared the 263K and 139A scrapie strains in transgenic mice expressing P101L PrP(C). Although the brains of 263K-affected mice had little immunoblot-detectable PrP(Res), RT-QuIC indicated that seeding activity was comparable to that associated with a high-PrP(Res) strain, 139A. Thus, in this comparison, RT-QuIC seeding activity correlated more closely with infectivity than with PrP(Res) levels. We also found that eQuIC, which incorporates a PrP(Sc) immunoprecipitation step, detected seeding activity in plasma from wild-type and anchorless PrP transgenic mice inoculated with 22L, 79A and/or RML scrapie strains. Overall, we conclude that these new mouse-adapted prion seeding assays detect diverse types of PrP(Sc).


A new device to study isoload eccentric exercise.
This study was designed to develop a new device allowing mechanical analysis of eccentric exercise against a constant load, with a view in mind to compare isoload (IL) and isokinetic (IK) eccentric exercises. A plate-loaded resistance training device was integrated to an IK dynamometer, to perform the acquisition of mechanical parameters (i.e., external torque, angular velocity). To determine the muscular torque produced by the subject, load torque was experimentally measured (TLexp) at 11 different loads from 30° to 90° angle (0° = lever arm in horizontal position). TLexp was modeled to take friction effect and torque variations into account. Validity of modeled load torque (TLmod) was tested by determining the root mean square (RMS) error, bias, and 2SD between the descending part of TLexp (from 30° to 90°) and TLmod. Validity of TLexp was tested by a linear regression and a Passing-Bablok regression. A pilot analysis on 10 subjects was performed to determine the contribution of the torque because of the moment of inertia to the amount of external work (W). Results showed the validity of TLmod (bias = 0%; RMS error = 0.51%) and TLexp SEM = 4.1 N·m; Intraclass correlation coefficient (ICC) = 1.00; slope = 0.99; y-intercept = -0.13). External work calculation showed a satisfactory reproducibility (SEM = 38.3 J; ICC = 0.98) and moment of inertia contribution to W showed a low value (3.2 ± 2.0%). Results allow us to validate the new device developed in this study. Such a device could be used in future work to study IL eccentric exercise and to compare the effect of IL and IK eccentric exercises in standardized conditions.


Veterinary pharmaceuticals and antibiotic resistance of Escherichia coli isolates in poultry litter from commercial farms and controlled feeding trials.
Veterinary pharmaceuticals are commonly used in poultry farming to prevent and treat microbial infections as well as to increase feed efficiency, but their use has created public and environmental health concerns. Poultry litter contains antimicrobial residues and resistant bacteria; when applied as fertilizer, the level and effects of these pharmaceuticals and antimicrobial-resistant bacteria in the environment are of concern. The purpose of this study was to investigate poultry litter for veterinary pharmaceuticals and resistance patterns of Escherichia coli. Litter samples were collected from controlled feeding trials and from commercial farms. Feed additives bacitracin, chlortetracycline, monensin, narasin, nicarbazin, penicillin, salinomycin, and virginiamycin, which were present in the feed on commercial farms and added to the feed in the controlled trials, were extracted in methanol and analyzed by liquid chromatography-mass spectrometry techniques. Sixty-nine E. coli were isolated and identified by API 20E. The susceptibility of the isolates to antibiotics was determined using Avian plates and the Sensititer automated system. This study confirmed the presence of antimicrobial residues in broiler litter from controlled environments as well as commercial farms, ranging from 0.07 to 66 mg/L depending on the compound. Concentrations of individual residues were higher in litter from controlled feeding trials than those from commercial farms. All E. coli isolates from commercial farms were multiresistant to at least 7 antibiotics. Resistance to beta-lactam antibiotics (amoxicillin, ceftiofur), tetracyclines, and sulfonamides was the most prevalent. This study concluded that broiler litter is a source of antimicrobial residues and represents a reservoir of multiple antibiotic-resistant E. coli.


Cooperative behavior in the workplace: Empirical evidence from the agent-deed-consequences model of moral judgment.
Moral judgment is of critical importance in the work context because of its implicit or explicit omnipresence in a wide range of work-place practices. The moral aspects of actual behaviors, intentions, and consequences represent areas of deep preoccupation, as exemplified in current corporate social responsibility programs, yet there remain ongoing debates on the best understanding of how such aspects of morality (behaviors, intentions, and consequences) interact. The ADC Model of moral judgment integrates the theoretical insights of three major moral theories (virtue ethics, deontology, and consequentialism) into a single model, which explains how moral judgment occurs in parallel evaluation processes of three different components: the character of a person (Agent-component); their actions (Deed-component); and the consequences brought about in the situation (Consequences-component). The model offers the possibility of overcoming difficulties encountered by single or dual-component theories. We designed a 2 × 2 × 2-between-subjects design vignette experiment with a Germany-wide sample of employed respondents (N = 1,349) to test this model. Results showed that the Deed-component affects willingness to cooperate in the work context, which is mediated via moral judgments. These effects also varied depending on the levels of the Agent- and Consequences-component. Thereby, the results exemplify the usefulness of the ADC Model in the work context by showing how the distinct components of morality affect moral judgment.


Metformin Inhibits Epithelial-to-Mesenchymal Transition of Keloid Fibroblasts via the HIF-1α/PKM2 Signaling Pathway.
Background: Epithelial-to-mesenchymal transition (EMT) is a process whereby epithelial cells lose cell-cell contacts and acquire expression of mesenchymal components and manifest a migratory phenotype. Recent studies indicated that EMT is involved in the development of keloids. Therefore, this study aims to investigate the mechanisms of the effects of metformin in hypoxia-induced EMT in keloid fibroblasts (KFs). Methods: KFs were cultured in a hypoxia incubator to induce EMT and were treated with or without metformin. Cell viability was evaluated by a cell counting kit 8 (CCK-8), and cell migration was measured by the transwell assay. The expression levels of HIF-1α, E-cadherin, vimentin, phosphorylated p70s6k (p-p70s6k) and pyruvate kinase M2 (PKM2) were evaluated by western blotting. Results: Hypoxia promoted EMT in KFs. Metformin significantly inhibited the expression of HIF-1α and partially abolished hypoxia-induced EMT. PKM2 is involved in hypoxia-induced EMT of KFs and metformin decreased the expression of p-p70s6k and PKM2. Conclusions: Metformin abolishes hypoxia-induced EMT in KFs by inhibiting the HIF-1α/PKM2 signaling pathway. Our study provides a novel mechanistic insight into potential use of metformin for treatment of keloids.


Fuzzy Evaluation Method for Haptic Perceptual Similarity.
Tactile rendering in virtual interactive scenes plays an important role in improving the quality of user experience. The subjective rating is currently the mainstream measurement to assess haptic rendering realism, which ignores various subjective and objective uncertainties in the evaluation process and also neglects the mutual influence among tactile renderings. In this paper, we extend the existing subjective evaluation and systematically propose a fuzzy evaluation method of haptic rendering realism. Hierarchical fuzzy scoring based on confidence interval is introduced to reduce the difficulty of expressing tactile feeling with deterministic rating. After the fuzzy statistics based on the membership function, we further use close-degree and transitive closure to calculate the fuzzy equivalence matrix between different tactile renderings. Fuzzy clustering is carried out to complete the comprehensive evaluation in the case of multiple indicators. Five tactile objects are used to simulate various situations of tactile rendering. The experimental results of haptic perceptual similarity evaluation show the existence of fuzziness in the subjective evaluation and verify the feasibility of the proposed method applied to multi-indicator evaluation. We also conclude that the proposed method outperforms the existing methods in terms of time cost and labor cost.


Effect of artificial dermal substitute, cultured keratinocytes and split thickness skin graft on wound contraction.
In this study, the effect of different wound treatments on contraction was evaluated in an established porcine model. In two separately conducted experiments full thickness wounds treated with artificial dermal substitute, split thickness skin graft (STSG), meshed STSG applied in combination with cultured keratinocytes or meshed STSG alone were compared with untreated wounds. The surface area of all wounds was quantified at regular time intervals. After 20 days wounds from some groups were subjected to histological analysis to establish the degree of epithelialization. Wounds treated with STSG contracted more than with artificial dermal substitute until day 21. From day 21 to day 35 wounds treated with STSG showed the least contraction. Wounds sprayed with cultured keratinocytes demonstrated a slower rate of contraction than those with meshed STSG alone after 20 days. The untreated control wounds showed a greater rate of contraction and had almost closed by day 20. This study demonstrates that there is a significant difference in contraction between wounds treated with artificial dermal substitute and control wounds and between wounds treated with STSG with cultured keratinocytes and meshed STSG alone. STSG with cultured keratinocytes, unmeshed STSG, and artificial dermal substitute all reduced wound contraction significantly.


Alfven waves in the solar corona.
Alfvén waves, transverse incompressible magnetic oscillations, have been proposed as a possible mechanism to heat the Sun's corona to millions of degrees by transporting convective energy from the photosphere into the diffuse corona. We report the detection of Alfvén waves in intensity, line-of-sight velocity, and linear polarization images of the solar corona taken using the FeXIII 1074.7-nanometer coronal emission line with the Coronal Multi-Channel Polarimeter (CoMP) instrument at the National Solar Observatory, New Mexico. Ubiquitous upward propagating waves were seen, with phase speeds of 1 to 4 megameters per second and trajectories consistent with the direction of the magnetic field inferred from the linear polarization measurements. An estimate of the energy carried by the waves that we spatially resolved indicates that they are too weak to heat the solar corona; however, unresolved Alfvén waves may carry sufficient energy.


Urinary excretion of fibrinogen-related materials, complement, and immunoglobulins in proliferative glomerulonephritis and after renal transplantation.
Using a radial immunodiffusion technique we measured the urinary concentration of material related to complement (C3), IgM, and IgG along with fibrin-fibrinogen degradation products and heterophile (sheep) haemagglutinins in 15 patients with proliferative glomerulonephritis and in 10 patients after renal transplantation. There was a significant correlation between all variables measured, and serial studies showed that with the exception of IgG-related materials potentially useful information could be obtained on the detection of rejection and the response to treatment in both conditions. The significance of these observations is discussed.


Delta9-tetrahydrocannabinol (Delta9-THC) prevents cerebral infarction via hypothalamic-independent hypothermia.
Delta(9)-tetrahydrocannabinol (Delta(9)-THC), a primary psychoactive constituent of cannabis, has been reported to act as a neuroprotectant via the cannabinoid CB(1) receptor. In this study, Delta(9)-THC significantly decreased the infarct volume in a 4 h mouse middle cerebral artery occlusion mouse model. The neuroprotective effect of Delta(9)-THC was completely abolished by SR141716, cannabinoid CB(1) receptor antagonist, and by warming the animals to 31 degrees C. Delta(9)-THC significantly decreased the rectal temperature, and the hypothermic effect was also inhibited by SR141716 and by warming to 31 degrees C. At 24 h after cerebral ischemia, Delta(9)-THC significantly increased the expression level of CB(1) receptor in both the striatum and cortex, but not in the hypothalamus. Warming to 31 degrees C during 4 h cerebral ischemia did not increase the expression of CB(1) receptor at the striatum and cortex in MCA-occluded mice. These results show that the neuroprotective effect of Delta(9)-THC is mediated by a temperature-dependent mechanism via the CB(1) receptor. In addition, warming to 31 degrees C might attenuate both the neuroprotective and hypothermic effects of Delta(9)-THC through inhibiting the increase in CB(1) receptor in both the striatum and cortex but not in the hypothalamus, which may suggest a new thermoregulation mechanism of Delta(9)-THC.


A New Method to Individualize Monitoring of Muscle Recovery in Athletes.
Assessment of muscle recovery is essential for the daily fine-tuning of training load in competitive sports, but individual differences may limit the diagnostic accuracy of group-based reference ranges. This article reports an attempt to develop individualized reference ranges using a Bayesian approach comparable to that developed for the Athlete Biological Passport. Urea and creatine kinase (CK) were selected as indicators of muscle recovery. For each parameter, prior distributions and repeated-measures SDs were characterized based on data of 883 squad athletes (1758 data points, 1-8 per athlete, years 2013-2015). Equations for the individualization procedure were adapted from previous material to allow for discrimination of 2 physiological states (recovered vs nonrecovered). Evaluation of classificatory performance was carried out using data from 5 consecutive weekly microcycles in 14 elite junior swimmers and triathletes. Blood samples were collected every Monday (recovered) and Friday according to the repetitive weekly training schedule over 5 wk. On the group level, changes in muscle recovery could be confirmed by significant differences in urea and CK and validated questionnaires. Group-based reference ranges were derived from that same data set to avoid overestimating the potential benefit of individualization. For CK, error rates were significantly lower with individualized classification (P vs group-based: test-pass error rate P = .008; test-fail error rate P < .001). For urea, numerical improvements in error rates failed to reach significance. Individualized reference ranges seem to be a promising tool to improve accuracy of monitoring muscle recovery. Investigating application to a larger panel of indicators is warranted.


In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein.
Previously, we identified that both fingers 1 and 2 in the three Cys2His2 zinc-finger domains (TZD) of testis zinc-finger protein specifically bind to its cognate DNA; however, finger 3 is non-sequence-specific. To gain insights into the interaction mechanism, here we further investigated the DNA-binding characteristics of TZD bound to non-specific DNAs and its finger segments bound to cognate DNA. TZD in non-specific DNA binding showed smaller chemical shift perturbations, as expected. However, the direction of shift perturbation, change of DNA imino-proton NMR signal, and dynamics on the 15N backbone atom significantly differed between specific and non-specific binding. Using these unique characteristics, we confirmed that the three single-finger segments (TZD1, TZD2 and TZD3) and the two-finger segment (TZD23) non-specifically bind to the cognate DNA. In comparison, the other two-finger segment (TZD12) binding to the cognate DNA features simultaneous non-specific and semi-specific binding, both slowly exchanged in terms of NMR timescale. The process of TZD binding to the cognate DNA is likely stepwise: initially TZD non-specifically binds to DNA, then fingers 1 and 2 insert cooperatively into the major groove of DNA by semi-specific binding, and finally finger 3 non-specifically binds to DNA, which promotes the specific binding on fingers 1 and 2 and stabilizes the formation of a specific TZD-DNA complex.


Trimethoprim-sulfamethoxazole for methicillin-resistant Staphylococcus aureus: a forgotten alternative?
Methicillin-resistant Staphylococcus aureus (MRSA) is a growing infectious concern, mainly in the context of its rapid adaptation to novel antibiotic options for its treatment and the growing morbidity, mortality, and healthcare costs associated with its emergence. the authors sought to investigate whether an older antibiotic, such as trimethoprim-sulfamethoxazole (SXT), may have a role in treating MRSA-related infections, according to the available literature on the subject. The authors reviewed literature data on: resistance of MRSA to SXT worldwide in recent years, efficacy of SXT for MRSA decolonization or prophylaxis from MRSA infections, and clinical therapeutic efficacy of SXT in treating mild or severe community-acquired or hospital-acquired MRSA infections. Resistance varies worldwide, in general being low in the industrialized world and higher in developing countries. SXT is one of the numerous understudied options for MRSA decolonization and is growingly recognized as potentially effective in preventing MRSA infections in certain settings. Limited data on its therapeutic efficacy are encouraging, at least for mild, community-acquired infections. SXT may represent a cost-effective alternative weapon against MRSA. Its utility against this increasingly threatening pathogen need clarification through further clinical trials.


Epidemiology and Management of Fusarium Wilt of China Asters.
The epidemiology and strategies for management of Fusarium wilt of China aster (Callistephus chinensis) were studied in Connecticut and Florida, USA, by examining seed contamination, on-farm disease incidence, sanitation, host resistance, and various soil treatments. Five out of 25 commercial seed packages from three separate distribution companies assayed in Connecticut had seeds contaminated with the pathogen Fusarium oxysporum f. sp. callistephi. Farm surveys of two cut-flower farms in Connecticut had disease incidences of 32 and 58%, while in Florida, the incidence of the disease ranged from 0.002 to 71.2% in two cut-flower operations. All pathogenic isolates from seed and symptomatic plants in Connecticut were vegetatively compatible, suggesting a common origin. Pathogenic isolates from Florida and nonpathogenic isolates fell into different vegetative compatibility groups and may have had another origin. Sodium hypochlorite solutions (1%) eliminated the fungus from seeds and Styrofoam when applied as a soak or spray, respectively. Soil fumigation with methyl bromide + chloropicrin, 1,3-dichloropropene + chloropicrin, or metam sodium maintained Fusarium wilt at low levels at a Florida cut-flower production facility. Evaluations of disease resistance of 44 cultivars in the greenhouse identified eight cultivars with moderate resistance. Four cultivars were identified with moderate resistance in field trials and thus could serve as a source of resistant germplasm for future breeding programs. These findings should encourage growers to use sanitation protocols to prevent entry of the pathogen into their fields and to choose commercially available cultivars that have moderate resistance.


Abdominal wall metastases following laparoscopy.
Only 18 cases of recurrence at the sites of cannula insertion after laparoscopy have been reported in the literature, ten of them in the past year. The period between laparoscopic surgery and presentation of wound metastasis varies widely, from 7 days to 10 months; the lesions are typically hard, craggy and painful. The most likely mechanism is direct implantation of viable exfoliated tumour cells but three aspects specific to laparoscopy may also be important. First, there may be increased exfoliation of tumour cells following manipulation by laparoscopic instruments of an unsuspected malignancy. Second, there may be repeated close contact between tumour-laden instruments and the port. Third, the passage of resected tissue through a small incision may coat the wound with potentially malignant cells.


The FDA alert on serotonin syndrome with combined use of SSRIs or SNRIs and Triptans: an analysis of the 29 case reports.
On July 19, 2006, the US Food and Drug Administration (FDA) issued an alert, "Potentially Life-Threatening Serotonin Syndrome With Combined Use of SSRIs or SNRIs and Triptan Medications." However, the cases that were the basis for the alert were not made available. The FDA recommends that patients treated concomitantly with a triptan and a selective serotonin reuptake inhibitor (SSRI)/selective norepinephrine reuptake inhibitor (SNRI) be informed of the possibility of serotonin syndrome. Following a Freedom of Information Act request, the FDA provided the 29 cases that they evaluated as the basis for the alert. I summarize the cases, rate the quality of the cases on the basis of the information provided, and then determine whether the cases fulfill the Sternbach and Hunter criteria for serotonin syndrome. Seven cases met the Sternbach criteria but did not meet the Hunter criteria. No cases met both criteria or just the Hunter criteria. Triptans when administered with SSRIs or SNRIs might rarely precipitate serotonin syndrome. The data do not support prohibiting the use of triptans with SSRIs or SNRIs. With increased physician awareness of serotonin syndrome, it is possible that additional cases may be reported.


Reducing preanalytical laboratory sample errors through educational and technological interventions.
A correct preanalytical phase procedure is critical to get an adequate sample and consequently to achieve the most reliable laboratory results, promoting patient safety. Continuous laboratory staff changes create the need to establish improvement strategies to reduce the error risk. The objective was to show how the numbers of preanalytical errors related to unsuitable samples in a hospital setting decrease following two improvement strategies related to new technology and educational actions and how their effects were measured by monitoring indicators. Samples were drawn by the laboratory and other hospital departments' nurses without previous patient appointment, therefore, prior tube preparation was not available before the phlebotomy. Corrective measures for these activities were established: educational program for nurses and a system of custom labels, which correlate each laboratory test in the Laboratory Information System (LIS) with the corresponding tube. Three phases were defined based on the implementation dates of the improvement actions to be assessed. The set of indicators designed to monitor the improvement related to clotted, hemolyzed, insufficient, and uncollected samples. Data were collected and indicators calculated from the LIS using a data warehouse application. Patient satisfaction with respect to phlebotomy was also measured annually using a scoring survey. There was a reduction in all types of preanalytical sample errors related to the improvement strategies adopted. The indicators demonstrated that the unavailable, insufficient, and clotted samples decreased between two- and three-fold, whereas hemolysis errors benefited more from these improvement strategies. Patient satisfaction with the laboratory and phlebotomy procedures improved over the past several years as based on the annual satisfaction surveys. The educational program for nursing personnel is relevant and important as can be seen in the decrease of sample errors and the resulting quality improvement. The custom label system minimizes the potential oversight of forgetting to draw a tube, which happens frequently when operating without appointments, by printing the labels according to requested tests. Detection, identification, and monitoring of the error and implementing strategies to improve preanalytical quality reduces error numbers and thereby improves patient safety and health system outcomes.


Mps1Mph1 Kinase Phosphorylates Mad3 to Inhibit Cdc20Slp1-APC/C and Maintain Spindle Checkpoint Arrests.
The spindle checkpoint is a mitotic surveillance system which ensures equal segregation of sister chromatids. It delays anaphase onset by inhibiting the action of the E3 ubiquitin ligase known as the anaphase promoting complex or cyclosome (APC/C). Mad3/BubR1 is a key component of the mitotic checkpoint complex (MCC) which binds and inhibits the APC/C early in mitosis. Mps1(Mph1) kinase is critical for checkpoint signalling and MCC-APC/C inhibition, yet few substrates have been identified. Here we identify Mad3 as a substrate of fission yeast Mps1(Mph1) kinase. We map and mutate phosphorylation sites in Mad3, producing mutants that are targeted to kinetochores and assembled into MCC, yet display reduced APC/C binding and are unable to maintain checkpoint arrests. We show biochemically that Mad3 phospho-mimics are potent APC/C inhibitors in vitro, demonstrating that Mad3p modification can directly influence Cdc20(Slp1)-APC/C activity. This genetic dissection of APC/C inhibition demonstrates that Mps1(Mph1) kinase-dependent modifications of Mad3 and Mad2 act in a concerted manner to maintain spindle checkpoint arrests.


[Effects of halotherapy on free radical oxidation in patients with chronic bronchitis].
Registration of luminol-dependent chemoluminescence of blood cells and iron-induced chemoluminescence of the serum was used to study generation of active oxygen forms and lipid peroxidation in patients with chronic bronchitis (CB). 49 patients with lingering CB showed inhibition of blood cell function and enhancement of lipid peroxidation. The addition of halotherapy to combined treatment of these patients promoted correction of the disorders and improvement of CB course.


Structural Analysis of the Effect of Asn107Ser Mutation on Alg13 Activity and Alg13-Alg14 Complex Formation and Expanding the Phenotypic Variability of ALG13-CDG.
Congenital Disorders of Glycosylation (CDG) are multisystemic metabolic disorders showing highly heterogeneous clinical presentation, molecular etiology, and laboratory results. Here, we present different transferrin isoform patterns (obtained by isoelectric focusing) from three female patients harboring the ALG13 c.320A>G mutation. Contrary to other known variants of type I CDGs, where transferrin isoelectric focusing revealed notably increased asialo- and disialotransferrin fractions, a normal glycosylation pattern was observed in the probands. To verify this data and give novel insight into this variant, we modeled the human Alg13 protein and analyzed the dynamics of the apo structure and the complex with the UDP-GlcNAc substrate. We also modeled the Alg13-Alg14 heterodimer and ran multiple simulations of the complex in the presence of the substrate. Finally, we proposed a plausible complex formation mechanism.


Risk factors for Alzheimer's disease: the EURODEM collaborative re-analysis of case-control studies.
Case-control studies of risk factors for Alzheimer's disease have been hampered by low statistical power. Here, we present a collaborative analysis of 11 case-control studies of Alzheimer's disease. The original data of these studies were pooled and re-analyzed. The risk factors considered in this analysis were family history of dementia and related disorders, maternal age at index birth, head trauma, medical and psychiatric history and environmental exposures. This paper gives a brief overview of the findings of the re-analysis.


Insights into eukaryotic DNA priming from the structure and functional interactions of the 4Fe-4S cluster domain of human DNA primase.
DNA replication requires priming of DNA templates by enzymes known as primases. Although DNA primase structures are available from archaea and bacteria, the mechanism of DNA priming in higher eukaryotes remains poorly understood in large part due to the absence of the structure of the unique, highly conserved C-terminal regulatory domain of the large subunit (p58C). Here, we present the structure of this domain determined to 1.7-A resolution by X-ray crystallography. The p58C structure reveals a novel arrangement of an evolutionarily conserved 4Fe-4S cluster buried deeply within the protein core and is not similar to any known protein structure. Analysis of the binding of DNA to p58C by fluorescence anisotropy measurements revealed a strong preference for ss/dsDNA junction substrates. This approach was combined with site-directed mutagenesis to confirm that the binding of DNA occurs to a distinctively basic surface on p58C. A specific interaction of p58C with the C-terminal domain of the intermediate subunit of replication protein A (RPA32C) was identified and characterized by isothermal titration calorimetry and NMR. Restraints from NMR experiments were used to drive computational docking of the two domains and generate a model of the p58C-RPA32C complex. Together, our results explain functional defects in human DNA primase mutants and provide insights into primosome loading on RPA-coated ssDNA and regulation of primase activity.


Determining Food Insecurity: An Application of the Rasch Model with Household Survey Data in Uganda.
The inexplicable nature of food insecurity in parts of Uganda and worldwide necessitated an investigation into the nature, extent, and differentials of household food security. The main objective of this study was to examine the food security dynamics and model household food insecurity. The Rasch modelling approach was employed on a dataset from a sample of 1175 (Tororo = 577; Busia = 598) randomly selected households in the year 2010. All households provided responses to the food security questions and none was omitted from the analysis. At 5 percent level of significance the analysis indicated that Tororo district average food security assessment (0.137 ± 0.181) was lower than that for Busia district (0.768 ± 0.177). All the mean square fit statistics were in the range of 0.5 to 1.5, and none of them showed any signs of distortion, degradation, or less productivity for measurement. This confirmed that items used in this study were very productive for measurement of food security in the study area. The study recommends further analysis where item responses are ordered polytomous rather than the dichotomous item response functions used. Furthermore, consideration should be given to fit models that allow for different latent distributions for households with children and those without children and possibly other subgroups of respondents.


The iron/heme regulated genes of Haemophilus influenzae: comparative transcriptional profiling as a tool to define the species core modulon.
Haemophilus influenzae requires heme for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. Although an understanding of the heme acquisition mechanisms of H. influenzae is emerging, significant gaps in our knowledge remain. Unresolved issues include the identities of all genes exhibiting altered transcription in response to iron and heme availability, the fraction of such genes functioning in iron/heme acquisition, and the heterogeneity of this gene set among clinical isolates. Previously we utilized H. influenzae strain Rd KW20 to demonstrate the utility of transcriptional profiling in defining the genes exhibiting altered transcription in response to environmental iron and heme levels. The current study expands upon those observations by determining the iron/heme modulons of two clinical isolates, the type b isolate 10810 and the nontypeable isolate R2866. These data are used to begin to define the core iron/heme modulon of the species. Microarray studies were performed to compare gene expression on transition from iron/heme-restricted to iron/heme-replete conditions for each isolate. Of 1820 ORFs on the array corresponding to R2866 genes, 363 were significantly differentially expressed: 233 were maximally transcribed under iron/heme-replete conditions and 130 under iron/heme-restricted conditions. Of the 1883 ORFs representing genes of strain 10810, 353 were significantly differentially transcribed: 150 were preferentially transcribed under iron/heme-replete conditions and 203 under iron/heme-restricted conditions. Comparison of the data sets indicated that 163 genes exhibited similar regulation in both isolates and that 74 of these exhibited similar patterns of regulation in Rd KW20. These comprise the putative core iron/heme modulon. This study provides evidence for a conserved core of H. influenzae genes the transcription of which is altered by the availability of iron and/or heme in the growth environment. Elucidation of this modulon provides a means to identify genes with unrecognized roles in iron/heme acquisition or homeostasis, unanticipated responsiveness to environmental levels of the micronutrients or potential roles in virulence. Defining these core genes is also of potential importance in identifying targets for therapeutic and vaccine designs since products of these genes are likely to be preferentially expressed during growth in iron/heme restricted sites of the human body.


A multicentre randomised double masked clinical trial of a new formulation of topical cysteamine for the treatment of corneal cystine crystals in cystinosis.
To evaluate the safety and efficacy of a new topical cysteamine formulation, stable at room temperature, for the treatment of corneal cystine crystals in cystinosis. 20 study subjects were enrolled in the safety study and 16 in the efficacy study. Both studies were randomised and double blind. The primary outcome for the safety study was the occurrence of predefined serious adverse reactions over 6 months and for the efficacy study the reduction of corneal cystine crystal score (CCCS) by 1.00 or more units on photographs graded by a reading centre using a standardised protocol. No study subject developed any serious adverse reactions. In the efficacy study, 47% of eyes receiving the standard formulation experienced a reduction in the CCCS of >/=1.00 after 1 year, while 7% of eyes on the new formulation experienced such a decrease (p=0.04). Although no serious adverse reactions were observed with either formulation, the new formulation was not as effective as the standard formulation.


Biotechnological production of taxol and related taxoids: current state and prospects.
Taxol is one of the most effective anti-cancer drugs ever developed. The natural source of taxol is the inner bark of several Taxus species, but it accumulates at a very low concentration and with a prohibitively high cost of extraction. Another problem is that the use of inner bark for taxol production implies the destruction of yew trees. For all these reasons, the growing demand for taxol greatly exceeds the supply that can be sustained by isolation from its natural source and alternative sources of the drug are being sought. Although taxol has been prepared by total synthesis, the process is not commercially viable. Taxol can also be semisynthetically produced via the conversion of baccatin III or 10-deacethylbaccatinIII found in Taxus needles but the cost and difficulty of the extraction process of the semisynthetic precursors are also very high. The most promising approach for the sustainable production of taxol and related taxoids is provided by plant cell cultures at an industrial level. Taxol is currently being clinically used against different tumour processes but due to the difficulty of its extraction and formulation, as well as the growing demand for the compound, new taxol analogues with improved properties are being studied. In this revision we discuss current research in the design of new taxol-related compounds, the chemical structure/anti-cancer activity relationship and new formulations of the drug. We also consider the optimizing strategies to improve taxol and related taxoid production in cell cultures, as well as the current knowledge of taxol metabolism, all of which are illustrated with examples, some of them from our own research.


Mitochondrial coupling factor 6 as a potent endogenous vasoconstrictor.
We demonstrated recently that coupling factor 6, an essential component of the energy-transducing stalk of mitochondrial ATP synthase, suppresses the synthesis of prostacyclin in vascular endothelial cells. Here, we tested the hypothesis that coupling factor 6 is present on the cell surface and is involved in the regulation of systemic circulation. This peptide is present on the surface of CRL-2222 vascular endothelial cells and is released by these cells into the medium. In vivo, the peptide circulates in the vascular system of the rat, and its gene expression and plasma concentration are higher in spontaneously hypertensive rats (SHRs) than in normotensive controls. Elevation of blood pressure with norepinephrine did not affect the plasma concentration of coupling factor 6. Intravenous injection of recombinant peptide increased blood pressure, apparently by suppressing prostacyclin synthesis, whereas a specific Ab to coupling factor 6 decreased systemic blood pressure concomitantly with an increase in plasma prostacyclin. Interestingly, the antibody's hypotensive effect could be abolished by treating with the cyclooxygenase inhibitor indomethacin. These findings indicate that mitochondrial coupling factor 6 functions as a potent endogenous vasoconstrictor in the fashion of a circulating hormone and may suggest a new mechanism for hypertension.


The microbiota, a necessary element of immunity.
The intestinal microbiota is essential for digestion, the production of physiological metabolites, and defense. More than 1013 bacteria are present in the intestine, inspiring awe as well as fear of potential infections. By definition, the immune system protects us from infection, and is given the task to recognize dangerous pathogens from useful mutualists. Nevertheless, the definition of pathogens and mutualists is often contextual, and the immune system reacts to all types of microbes. In fact, immune reactivity to microbiota is necessary for the development of the immune system. If the host-microbe cross-talk is perturbed before birth or weaning, the immune system develops "pathological imprinting" and increased susceptibility to inflammatory pathology later in life. Reactivity to microbiota is also important in adulthood to regulate immune responses and maintain homeostasis.


Increased risk for urological cancer associated with anxiety disorder: a retrospective cohort study.
Anxiety disorders (ADs) are common with a high rate of medical comorbidities. Although the association between ADs and the overall cancer risk remains controversial, patients with ADs were found to be more likely to develop specific cancer types. Herein, we estimated the risk of developing urological cancers among patients with ADs in a 5-year follow-up period using a population-based database. Two study cohorts were identified from the Taiwan Longitudinal Health Insurance Database 2005: patients with ADs, and comparison subjects selected by one-to-one matching for sex, age, and the year of recruitment. Follow-up was undertaken to determine whether sampled patients and comparison subjects had developed urological cancers in the subsequent 5 years. We found that urological cancers occurred among 0.54% of patients with ADs and 0.13% of comparison subjects. After adjusting for sociodemographic characteristics, medical comorbidities, and alcohol and tobacco use disorder, the stratified Cox proportional hazard regression suggested that patients with ADs were more likely to develop urological cancers relative to comparison subjects (adjusted hazard ratio, 3.67; 95% confidence interval, 2.85 ~ 4.72). The adjusted HR for males with ADs was 3.82 (95% CI: 2.79 ~ 5.23) in comparison to males without ADs. In addition, the adjusted HR for females with ADs was 3.47 (95% CI: 2.26 ~ 5.31) than those females without ADs. We concluded that during the 5-year follow-up period, there was a significantly increased risk of urological cancers among patients with ADs.


DEEP CLUSTERING AND CONVENTIONAL NETWORKS FOR MUSIC SEPARATION: STRONGER TOGETHER.
Deep clustering is the first method to handle general audio separation scenarios with multiple sources of the same type and an arbitrary number of sources, performing impressively in speaker-independent speech separation tasks. However, little is known about its effectiveness in other challenging situations such as music source separation. Contrary to conventional networks that directly estimate the source signals, deep clustering generates an embedding for each time-frequency bin, and separates sources by clustering the bins in the embedding space. We show that deep clustering outperforms conventional networks on a singing voice separation task, in both matched and mismatched conditions, even though conventional networks have the advantage of end-to-end training for best signal approximation, presumably because its more flexible objective engenders better regularization. Since the strengths of deep clustering and conventional network architectures appear complementary, we explore combining them in a single hybrid network trained via an approach akin to multi-task learning. Remarkably, the combination significantly outperforms either of its components.