[3H]GBR 12935 binding in platelets from poor and extensive cytochrome P-4502D6 metabolizers.
Previous studies have indicated that part of the binding of [3H] [1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl) piperazine dihydrochloride] ([3H]GBR 12935) to human platelets is to a piperazine acceptor site, which might be associated with cytochrome P-450IID6 (CYP4502D6, debrisoquine-4-hydroxylase). Due to mutant CYP4502D6 alleles, 5-10% of Caucasians are poor metabolizers of CYP4502D6 substrates such as debrisoquine and dextromethorphan. In the present study, possible differences in binding characteristics of [3H]GBR 12935 in platelets from CYP4502D6 poor and extensive metabolizers were investigated. The most prominent finding was a gender difference, with males having significantly higher Kd values than females. There were no differences in Bmax. After correction for gender, there was a tendency towards higher Kd values in poor metabolizers than in extensive metabolizers, although the difference was not statistically significant. Whether this finding corresponds to reduced CYP4502D6 activity is a matter of further investigation.


Purple urine bag syndrome: case report and literature review.
Purple urine bag syndrome (PUBS) is a medical syndrome in which there is purple discoloration of the urine of catheterized patients as well as discoloration of the collecting bag and the associated tubing. This rare condition, which mostly affects women, is generally associated with catheter-associated urinary tract infection, chronic constipation and alkaline urine. PUBS may be caused by sequential chemical reactions involving tryptophan from food in the gastrointestinal tract. The clinical course of PUBS is generally benign, and intensive treatment is not usually needed. We present 3 cases of this unusual and interesting phenomenon and a literature review.


Polymer monoliths as efficient solid phases for enzymatic polynucleotide degradation followed by fast HPLC analysis.
Two ribonuclease A bioreactors based on lab-made macroporous monolithic columns and intended for polynucleotide degradation were prepared using in situ free-radical polymerization. Different methods of enzyme immobilization were applied. In the first case, the biocatalyst molecule was attached to the solid surface via direct covalent binding, while in the second bioreactor the flexible-chain synthetic polymer was used as an intermediate spacer. The effect of temperature, substrate flow rate, and loaded sample volume on the biocatalytic efficiency of the immobilized enzyme was examined. The kinetic parameters of the enzymatic degradation of synthetic polycytidylic acid were calculated and compared to those found for hydrolysis with soluble ribonuclease A. The monitoring of substrate splitting was carried out by means of fast anion-exchange HPLC on an ultra-short monolithic column (disk) using off- and on-line analytical approaches.


Evaluating the effect of state regulation of federally licensed firearm dealers on firearm homicide.
Effective federal regulation of firearm dealers has proven difficult. Consequently, many states choose to implement their own regulations. We examined the impact of state-required licensing, record keeping of sales, allowable inspections, and mandatory theft reporting on firearm homicide from 1995 to 2010. We found that lower homicide rates were associated with states that required licensing and inspections. We concluded that firearm dealer regulations might be an effective harm reduction strategy for firearm homicide.


An investigation of preschool teachers' recognition of possible child abuse and neglect in Izmir, Turkey.
Child abuse and neglect have a potentially deleterious impact on children's physical, social, and psychological development. Preschool teachers may play a crucial role in the protection, early detection, and the intervention of child abuse and neglect, as they have the opportunity to establish a close contact with the families and to observe day-to-day changes in pupils' behavior. The main purpose of this study is to investigate preschool teachers' experiences and characteristics in relation to their awareness of possible child abuse and neglect signs. A questionnaire survey was designed and administered to 197 preschool teachers who work for the public preschools in the Izmir province of Turkey. In addition to the questionnaire items, a 34-item Likert-type scale measuring the level of familiarity with possible signs of child abuse and neglect was developed. This scale had an internal consistency of 0.94. The results revealed that 10.65% of preschool teachers had training regarding violence against children and 2.03% of them had training in child abuse and neglect. Overall, 35% of all teachers reported that they had prior experience with pupils who were exposed to child abuse and neglect. Moreover, statistical analyses indicated that being a parent and having training in child abuse and neglect, having experience with maltreated children, and having higher job status were significant factors in preschool teachers' ability to recognize the possible signs of child abuse and neglect. Our results support that teacher training in child abuse and neglect can play an important role in preschool teachers' awareness of the possible signs of child abuse and neglect.


Long-lasting inhibition of the cardiovascular responses to glutamate and the baroreceptor reflex elicited by neuropeptide Y injected into the nucleus tractus solitarius of the rat.
Neuropeptide Y (NPY) microinjected unilaterally into the nucleus tractus solitarii (NTS) of anesthetized paralyzed rats elicits a gradual dose-dependent and reversible fall in arterial pressure (AP) and heart rate (HR) lasting 20 min. It also abolished the brief (less than 1 min) dose-dependent and reversible fall of AP and HR elicited by L-glutamate (L-Glu) injected into the nucleus. The blockade of L-Glu by NPY appeared gradually and was prolonged, lasting over 2 h, and recovering by 24 h. It was not replicated by desamido-NPY or galanin. Unlike 2% lidocaine it did not block the hypotension elicited by focal electrical stimulation at the injection site indicating the response was not that of a local anesthetic. Bilateral injection of NPY into the NTS resulted, after an initial fall, in an elevation of AP (+48 +/- 10.6 mmHg). At this time the reflex bradycardia evoked by elevating AP with phenylephrine was markedly reduced. We conclude that in the NTS, NPY antagonizes the actions of L-Glu and may attenuate baroreceptor reflexes. Since the NTS is richly innervated by NPY neurons and contains many NPY binding sites and since primary baroreceptor afferents appear to be glutamatergic the results suggested that NPY may serve in NTS as a long-term regulator of baroreceptor reflex activity.


Bioactive Compounds and Bioactivities of Ginger (Zingiber officinale Roscoe).
Ginger (Zingiber officinale Roscoe) is a common and widely used spice. It is rich in various chemical constituents, including phenolic compounds, terpenes, polysaccharides, lipids, organic acids, and raw fibers. The health benefits of ginger are mainly attributed to its phenolic compounds, such as gingerols and shogaols. Accumulated investigations have demonstrated that ginger possesses multiple biological activities, including antioxidant, anti-inflammatory, antimicrobial, anticancer, neuroprotective, cardiovascular protective, respiratory protective, antiobesity, antidiabetic, antinausea, and antiemetic activities. In this review, we summarize current knowledge about the bioactive compounds and bioactivities of ginger, and the mechanisms of action are also discussed. We hope that this updated review paper will attract more attention to ginger and its further applications, including its potential to be developed into functional foods or nutraceuticals for the prevention and management of chronic diseases.


The 2009 influenza pandemic: promising lessons for antiviral therapy for future outbreaks.
The influenza A virus is the main circulating influenza virus in the human population. It can cause disease also in birds and other mammals and is responsible for annual epidemics and occasional pandemics. The most known and deadly pandemic was the "Spanish flu" (influenza type A/H1N1), which struck the human population between 1918 and 1919, with probably the heaviest toll ever recorded in terms of human lives. The most recent flu pandemic, caused in 2009 by the swine-origin reassortant virus (pH1N1), has raised several critical issues in terms of our preparedness in responding fast to new pandemic influenza strains. Probably, the most instructive lesson that has been learned from the 2009 pandemic, was that the speed of manufacturing and distributing an effective vaccine will not be able to keep up with the pace of a rapidly spreading pandemic virus, failing to grant accessibility to the vaccine for a significant percentage of the susceptible population, before the onset of the pandemic peak. Thus, our first and most effective line of defense against a pandemic influenza virus, particularly in the early phases, are the antiviral drugs. Here we analyze our current understanding of the influenza pandemic viruses, in general, and of the pH1N1 in particular, along with the most recent approaches being pursued to design new anti-influenza drugs.


Recommendations for VTE Prophylaxis in Medically Ill Patients.
Venous thromboembolism (VTE) remains highly prevalent in medically ill patients, and often leads to increased mortality and cost burden during hospitalization and post-discharge. Nearly half of all VTEs occur during or after hospitalization, with pulmonary embolism accounting for 10% of inpatient mortality. Appropriate prophylaxis in high-risk medically ill patients has been shown to reduce risk of VTE and related mortality. Despite current evidence-based guidelines, VTE prophylaxis has been under-used. This owes greatly to ambiguity and concerns related to appropriate patient and prophylactic agent selection, and duration of prophylaxis. Because many acutely ill medical patients have multiple comorbidities, the risk of major bleeding must be considered when choosing to implement pharmacological VTE prophylaxis. Multiple risk assessment models have been developed and validated to help estimate VTE and bleeding risks in this population. While studies have shown that the risk for VTE often extends far beyond hospital discharge, there is no evidence to support extending prophylaxis after hospital discharge. The appropriate selection of VTE prophylaxis requires consideration for cost, availability, patient preference, compliance, and underlying comorbidities. Our paper reviews the current evidence and reasoning for appropriate selection of VTE prophylaxis in acutely medical ill patients, and highlights our own approach and recommendations.


Childhood immune thrombocytopenia: role of rituximab, recombinant thrombopoietin, and other new therapeutics.
Childhood immune thrombocytopenia (ITP) is often considered a benign hematologic disorder. However, 30% of affected children will have a prolonged course and 5%-10% will develop chronic severe refractory disease. Until recently, the only proven therapeutic option for chronic severe ITP was splenectomy, but newer alternatives are now being studied. However, because immunosuppressive agents such as rituximab are not approved for use in ITP and the thrombopoietin receptor agonists are not yet approved in children, the decision to use alternatives to splenectomy needs to be considered carefully. This review describes the factors that should affect decisions to treat ITP at diagnosis and compares the options for the occasional child in whom ITP does not resolve within the first year.


Impact of treatment success on health service use and cost in depression: longitudinal database analysis.
Research has consistently demonstrated a relationship between depression and increased levels of health service use over the short term. However, much less is known about how this relationship is influenced by the success, or otherwise, of depression management strategies, and the economic impact over the longer term. To investigate the economic impact of non-remission on health service use and costs over 12 months from the index episode in patients with depression. A naturalistic, longitudinal study was carried out using data from a large primary care UK general practice research database between 2001 and 2006. The records of 88 935 patients aged ≥18 years, diagnosed with depression and in receipt of at least two antidepressant prescriptions (for amitriptyline, citalopram, escitalopram, fluoxetine, paroxetine, sertraline or venlafaxine) in the first 3 months after the index prescription were included. The main outcome measures were health service use and cost over the 12-month study period, by remission status, where remission is defined as patients not using antidepressants for at least 6 months after antidepressant treatment has ended. Sociodemographic and clinical characteristics were similar for participants classified as in remission and those not in remission. Over 12 months from the index prescription, patients classified as non-remitters spent longer, on average, than patients classified as remitters on concomitant psychotropic medication (204 days vs 93 days, respectively), and had more contact with primary care services (17 vs 13 GP visits), secondary care psychiatrists and other specialists (47% vs 40%). Days in hospital, accident and emergency attendances and psychological therapy contacts did not differ between the groups. Total 12-month costs per participant were significantly lower for remitters (mean £656 vs £937; mean difference £317; p < 0.0001). Total costs fell over time for both groups, but at a faster rate for those in remission, and for those who remitted earlier after the index prescription than for those who remitted later. Successful cessation of antidepressant medication treatment in adults with depression can result in significant cost savings to the health service.


Loss of nonsense mediated decay suppresses mutations in Saccharomyces cerevisiae TRA1.
Tra1 is an essential protein in Saccharomyces cerevisiae. It was first identified in the SAGA and NuA4 complexes, both with functions in multiple aspects of gene regulation and DNA repair, and recently found in the ASTRA complex. Tra1 belongs to the PIKK family of proteins with a C-terminal PI3K domain followed by a FATC domain. Previously we found that mutation of leucine to alanine at position 3733 in the FATC domain of Tra1 (tra1-L3733A) results in transcriptional changes and slow growth under conditions of stress. To further define the regulatory interactions of Tra1 we isolated extragenic suppressors of the tra1-L3733A allele. We screened for suppressors of the ethanol sensitivity caused by tra1-L3733A. Eleven extragenic recessive mutations, belonging to three complementation groups, were identified that partially suppressed a subset of the phenotypes caused by tra1-L3733A. Using whole genome sequencing we identified one of the mutations as an opal mutation at tryptophan 165 of UPF1/NAM7. Partial suppression of the transcriptional defect resulting from tra1-L3733A was observed at GAL10, but not at PHO5. Suppression was due to loss of nonsense mediated decay (NMD) since deletion of any one of the three NMD surveillance components (upf1/nam7, upf2/nmd2, or upf3) mediated the effect. Deletion of upf1 suppressed a second FATC domain mutation, tra1-F3744A, as well as a mutation to the PIK3 domain. In contrast, deletions of SAGA or NuA4 components were not suppressed. We have demonstrated a genetic interaction between TRA1 and genes of the NMD pathway. The suppression is specific for mutations in TRA1. Since NMD and Tra1 generally act reciprocally to control gene expression, and the FATC domain mutations do not directly affect NMD, we suggest that suppression occurs as the result of overlap and/or crosstalk in these two broad regulatory networks.


Cholinergic modulation of neocortical long-term potentiation in the awake, freely moving rat.
The neocortex has proven resistant to LTP induction using standard in vitro and acute, in vivo preparations. Because the neocortex is widely thought to be involved in long-term information storage, this resistance raises questions about the validity of LTP as a memory model. Recently, we have shown that the neocortex of freely moving rats reliably supports LTP, provided that the stimulation is spaced and repeated over days. The following experiments were designed to evaluate the neuromodulatory role played by cholinergic systems in the induction of LTP in this preparation. Chronically implanted rats received either low- or high-intensity LTP-inducing tetani in combination with the administration of either a cholinergic agonist or antagonist injected systemically. Potentiation was evidenced as amplitude changes in both early and late components of the evoked field potential, the former including population spikes. The cholinergic agonist facilitated LTP induction in the late component of both high- and low-intensity groups. The cholinergic antagonist blocked LTP induction in the early component of the high-intensity group. The possibility that there are component-specific modulatory effects of cholinergic agents on the induction of neocortical LTP is discussed.


Challenges in measuring outcomes for caregivers of people with mental health problems.
Patient-reported outcomes (PROs) are increasingly important in health care and mental health research. Furthermore, caregivers become partners in care for patients with mental disorders, and health workers are more attentive to the expectations and needs of caregivers. A number of outcomes for caregivers are measured and used in daily practice in order to promote actions to improve health care systems and progress in research on the impact of mental disorders on their caregivers. This paper proposes an inventory of the different outcomes and different measurement tools used to assess the impact of disorders, raising a number of methodological and conceptual issues that limit the relevance of measurement tools and complicate their use. Finally, we propose some recommendations promoting the development of relevant outcome measures for caregivers and their integration into current systems of care.


CaO2-Cu2O micromotors accelerate infected wound healing through antibacterial functions, hemostasis, improved cell migration, and inflammatory regulation.
During the wound tissue healing process, the relatively weak driving forces of tissue barriers and concentration gradients lead to a slow and inefficient penetration of bioactive substances into the wound area, consequently showing an impact on the effectiveness of deep wound healing. To overcome these challenges, we constructed biocompatible CaO2-Cu2O "micromotors". These micromotors reacted with the fluids at the wound site, releasing oxygen bubbles and propelling particles deep into the wound tissue. In vitro experimental results revealed that these micromotors not only exhibited antibacterial and hemostatic functions but also facilitated the migration of dermal fibroblasts and vascular endothelial cells, while modulating the inflammatory microenvironment. A methicillin-resistant Staphylococcus aureus infected full-thickness-wound model was created in rats, in which CaO2-Cu2O micromotors markedly expedited the wound healing process. Specifically, CaO2-Cu2O provided a sterile microenvironment for wounds and increased the amounts of M1-type macrophages during infection and inflammation. During the proliferation and remodeling stages, the amount of M1 macrophages gradually decreased, while the amount of M2 macrophages increased, and CaO2-Cu2O did not prolong the inflammatory period. Furthermore, the introduction of a regenerated silk fibroin (RSF) film on the wound surface successfully enhanced the therapeutic effects of CaO2-Cu2O against the infected wound. The combined application of oxygen-producing CaO2-Cu2O micromotors and a RSF film demonstrates significant therapeutic potential and emerges as a promising candidate for the treatment of infected wounds.


BITC Sensitizes Pancreatic Adenocarcinomas to TRAIL-induced Apoptosis.
Pancreatic adenocarcinoma is an aggressive cancer with a greater than 95% mortality rate and short survival after diagnosis. Chemotherapeutic resistance hinders successful treatment. This resistance is often associated with mutations in codon 12 of the K-Ras gene (K-Ras 12), which is present in over 90% of all pancreatic adenocarcinomas. Codon 12 mutations maintain Ras in a constitutively active state leading to continuous cellular proliferation. Our study determined if TRAIL resistance in pancreatic adenocarcinomas with K-Ras 12 mutations could be overcome by first sensitizing the cells with Benzyl isothiocyanate (BITC). BITC is a component of cruciferous vegetables and a cell cycle inhibitor. BxPC3, MiaPaCa2 and Panc-1 human pancreatic adenocarcinoma cell lines were examined for TRAIL resistance. Our studies show BITC induced TRAIL sensitization by dual activation of both the extrinsic and intrinsic apoptotic pathways.


Assembly of AUF1 with eIF4G-poly(A) binding protein complex suggests a translation function in AU-rich mRNA decay.
An AU-rich element (ARE) located in the 3'-untranslated region of many short-lived mRNAs functions as an instability determinant for these transcripts. AUF1/hnRNP D, an ARE-binding protein family consisting of four isoforms, promotes rapid decay of ARE-mRNAs. The mechanism by which AUF1 promotes rapid decay of ARE-mRNA is unclear. AUF1 has been shown to form an RNase-resistant complex in cells with the cap-initiation complex and heat shock proteins Hsp70 and Hsc70, as well as other unidentified factors. To understand the function of the AUF1 complex, we have biochemically investigated the association of AUF1 with the components of the translation initiation complex. We used purified recombinant proteins and a synthetic ARE RNA oligonucleotide to determine the hierarchy of protein interactions in vitro and the effect of AUF1 binding to the ARE on the formation of protein complexes. We demonstrate that all four AUF1 protein isoforms bind directly and strongly to initiation factor eIF4G at a C-terminal site regardless of AUF1 interaction with the ARE. AUF1 is shown to directly interact with poly(A) binding protein (PABP), both independently of eIF4G and in a complex with eIF4G. AUF1-PABP interaction is opposed by AUF1 binding to the ARE or Hsp70 heat shock protein. In vivo, AUF1 interaction with PABP does not alter PABP stability. Based on these and other data, we propose a model for the molecular interactions of AUF1 that involves translation-dependent displacement of AUF1-PABP complexes from ARE-mRNAs with possible unmasking of the poly(A) tail.


Sialoadhesin (CD169) expression in CD14+ cells is upregulated early after HIV-1 infection and increases during disease progression.
Sialoadhesin (CD169, siglec-1 or Sn) is an activation marker seen on macrophages in chronic inflammatory diseases and in tumours, and on subsets of tissue macrophages. CD169 is highly expressed by macrophages present in AIDS-related Kaposi's sarcoma lesions. It is also increased on blood monocytes of HIV-1 infected patients with a high viral load despite antiretroviral treatment. We investigated expression of sialoadhesin in untreated HIV-1 and HHV-8 infected patients, by real-time PCR and FACS analysis to establish its expression in relation to infection and disease progression. Patients analysed were either HIV-1 seroconverters (n = 7), in the chronic phase of HIV-1 infection (n = 21), or in the AIDS stage (n = 58). Controls were HHV-8 infected, but otherwise healthy individuals (n = 20), and uninfected men having sex with men (n = 24). Sialoadhesin mRNA was significantly elevated after HIV-1, but not HHV-8 infection, and a further increase was seen in AIDS patients. Samples obtained around HIV-1 seroconversion indicated that sialoadhesin levels go up early in infection. FACS analysis of PBMCs showed that sialoadhesin protein was expressed at high levels by approximately 90% of CD14(+) and CD14(+)CD16(+)cells of HIV-1(+) patients with a concomitant 10-fold increase in sialoadhesin protein/cell compared with uninfected controls. We have shown that sialoadhesin is induced to high levels on CD14(+) cells early after HIV-1 infection in vivo. The phenotype of the cells is maintained during disease progression, suggesting that it could serve as a marker for infection and probably contributes to the severe dysregulation of the immune system seen in AIDS.


The silent treatment: parents' narratives of sexuality education with young people.
This paper is based on research undertaken in Ireland that sought to understand how parents communicate with their children about sexuality. Forty-three parents were interviewed and data were analysed using analytical induction. Data indicated that while parents tended to pride themselves on the culture of openness to sexuality that prevailed in their home, they often described situations where very little dialogue on the subject actually transpired. However, unlike previous research on the topic that identified parent-related factors (such as ignorance or embarrassment) as the main impediments to parent-young person communication about sex, participants in our study identified the central obstacle to be a reticence on the part of the young person to engage in such dialogue. Participants described various blocking techniques apparently used by the young people, including claims to have full prior knowledge on the issue, physically absenting themselves from the situation, becoming irritated or annoyed, or ridiculing parents' educational efforts. In our analysis, we consider our findings in light of the shifting power of children historically and the new cultural aspiration of maintaining harmonious and democratic relations with one's offspring.


The La protein from human liver cells interacts specifically with the U-rich region in the hepatitis C virus 3' untranslated region.
To identify cellular factors in human liver that interact with hepatitis C virus (HCV) 5' and 3' untranslated regions (UTRs) and therefore possibly are involved in the regulation of HCV transcription or translation. We prepared cytoplasmic extracts from human liver biopsy samples and fractionated cellular factors on ion exchange columns. The various fractions from ion exchange columns were subjected to ultraviolet (UV) cross-linking with radiolabeled HCV 5' and 3' UTR RNA probes. Two major 45- and 46-kd proteins that interacted specifically with the HCV 5' and 3' UTR were identified. Using Western blot and immunoprecipitation, these proteins were identified as the La antigen. Our results demonstrate that the La protein from human liver biopsy cells interacts specifically with the U-rich region in the positive strand of the HCV 3' UTR.


A Prospective Study of Altered Inflammatory Response and Its Clinical Outcome following Laparoscopic and Open Cholecystectomy.
Inflammatory response following surgical trauma has long been a matter of study. Results, however, have been varied. We sought to assess changes in the levels of proinflammatory and anti-inflammatory cytokines in patients undergoing laparoscopic and open cholecystectomy and their impact on the clinical outcome of patients concerning the postoperative pain score. The study involved 90 cholecystectomies (55 laparoscopic and 35 open) for chronic cholecystitis. Blood samples were collected 2 hours preoperatively and at 4 and 24 hours post surgery. Sera were evaluated for the levels of interleukin-1β, interleukin-10, and tumor necrosis factor-alpha. The independent sample t-test was used to compare the means of a variable between the 2 groups. Statistical analysis was done using SPSS, version 17. The rise in the levels of interleukin-1β, interleukin-10, and tumor necrosis factor-alpha was significantly more in the open cholecystectomy group at 4 hours (P<0.00). At the 24th postoperative hour, the levels of all 3 cytokines were also higher in the open cholecystectomy group (P<0.001 for interleukin-1β, P=0.185 for interleukin-10, and P<0.001 for tumor necrosis factor-alpha). At the 4th postoperative hour, the patients in the laparoscopic cholecystectomy group had a significantly lower pain score (P<0.001) than the open group. Both laparoscopic and open cholecystectomy procedures altered the inflammatory milieu of our patients in the postoperative period. Inflammation caused by the laparoscopic procedure was significantly less. More research is needed to target specific inflammatory and anti-inflammatory cytokines to reduce surgical stress and improve patient outcomes.


Detection of QTLs for Stagonospora glume blotch resistance in Swiss winter wheat.
Stagonospora nodorum is the causal agent of the Stagonospora glume blotch disease in hexaploid wheat. The Swiss winter bread wheat cv. 'Arina' has a highly effective, durable and quantitative glume blotch resistance. We studied 240 single seed descent (SSD)-derived lines of an 'Arina x Forno' F(5:7) population to identify and map quantitative trait loci (QTLs) for glume blotch resistance under natural infestation. Using composite interval mapping (CIM) and LOD>4.5, we detected two chromosomal regions on chromosome arms 3BS and 4BL which were specifically associated with glume blotch resistance. These identified QTLs were designated QSng.sfr-3BS and QSng.sfr-4BL, respectively. QSng.sfr-3BS peaked at the locus Xgwm389 in the telomeric region of the short arm of chromosome 3B and explained 31.2% of the observed phenotypic variance for the resistance within the population. The responsible QSng.sfr-3BS allele originated from the resistant parent 'Arina'. The QTL QSng.sfr-4BL (19.1%) mapped to chromosome arm 4BL ('Forno' allele) very close to two known genes, TaMlo and a catalase ( Cat). Both QTL alleles combined could enhance the resistance level by about 50%. Additionally, they showed significant epistatic effects (4.4%). We found PCR-based microsatellite markers closely linked to QSng.sfr-3BS (gwm389) and QSng.sfr-4BL (gwm251) which make marker-assisted selection (MAS) for Stagonospora glume blotch resistance feasible. We also found one resistance QTL, QSng.sfr-5BL, on the long arm of chromosome 5B which overlapped with QTLs for plant height as well as heading time.


Mycotic aneurysm in a turtle hunter: brief review and a case report.
Salmonella-associated mycotic aneurysm is a rare, but dreaded, complication of salmonellosis. Immunocompromised and elderly populations are more susceptible to develop this extra-intestinal complication. Salmonella is spread via fecal-oral and vehicle-borne routes. Reptiles, especially small pet turtles, have been linked with an increased risk of Salmonella infection. Diagnosis of mycotic aneurysm is a challenge due to atypical presentations. Recently, widespread use of CT scan imaging to evaluate for unexplained abdominal pain and sepsis has led to early identification of mycotic aneurysms. Antibiotic therapy and surgical intervention are the cornerstones of management. Open surgery has been the gold standard of treatment but is associated with increased morbidity and mortality. A relatively new alternative to open surgery is endovascular aneurysm repair (EVAR). It is comparatively less invasive and is associated with reduced early morbidity and mortality in the setting of mycotic aneurysm. However, there is a risk of late infection. Here, we present a patient with Salmonella mycotic aneurysm initially treated conservatively with antibiotic therapy who later underwent successful interval EVAR with no complications to date. Also included is a brief review of Salmonella-associated mycotic aneurysms.


Study of heart-reactive antibody in antisera and hybridoma culture fluids against group A streptococci.
Heart-reactive antibody is known to be produced in animals immunized with group A streptococci. However, the detection and quantitation of these antibodies and their respective antigens have been limited to immunofluorescence or immunoprecipitation tests. In this study a more sensitive technique was used to detect heart-reactive antibody, i.e., the enzyme-linked immunosorbent assay (ELISA). Countercurrent immunoelectrophoresis and agar gel immunodiffusion were also used to confirm the reactivity of rabbit antisera to group A streptococci and human heart tissue. Human heart tissue antigen was prepared by Triton X-100 extraction, and checkerboard titrations of heart tissue antigen versus streptococcal antisera revealed optimal concentrations of each for the ELISA. When immune rabbit serum was reacted with heart tissue antigen, a 5- to 10-fold increase was observed over the reactions of preimmune sera controls. Streptococcal antiserum was found to have a six- to eightfold greater reactivity with heart tissue antigen than with similar concentrations of kidney and skeletal muscle antigens. Heart tissue antigen was partially purified by DEAE-cellulose chromatography, and quantities of greater than 10 to 100 ng (dry weight) were shown to react with streptococcal antisera in the ELISA. Heart, kidney, and skeletal muscle antigens were subjected to electrophoresis in polyacrylamide gel slabs and blotted onto a nitrocellulose filter. These filter blots reacted with streptococcal antisera and confirmed the tissue antigen specificity observed with the ELISA. In addition, the ELISA was found to be an effective method for the detection of heart-reactive antibodies produced by murine hybridomas that were producing antibody to group A streptococci.


Phenolic and carotenoid profiles of papaya fruit (Carica papaya L.) and their contents under low temperature storage.
Tropical fruits are rich in phenolic and carotenoid compounds, and these are associated with cultivar, pre- and postharvest handling factors. The aim of this work was to identify major phenolics and carotenoids in 'Maradol' papaya fruit and to investigate their response to storage temperature. Ferulic acid, caffeic acid and rutin were identified in 'Maradol' papaya fruit exocarp as the most abundant phenolic compounds, and lycopene, β-cryptoxanthin and β-carotene were identified in mesocarp as the major carotenoids. Ranges of contents of ferulic acid (1.33-1.62 g kg(-1) dry weight), caffeic acid (0.46-0.68 g kg(-1) dw) and rutin (0.10-0.16 g kg(-1) dw) were found in papaya fruit, which tend to decrease during ripening at 25 °C. Lycopene (0.0015 to 0.012 g kg(-1) fresh weight) and β-cryptoxanthin (0.0031 to 0.0080 g kg(-1) fw) were found in fruits stored at 25 °C, which tend to increase during ripening. No significant differences in β-carotene or rutin contents were observed in relation to storage temperature. Phenolics and carotenoids of 'Maradol' papaya were influenced by postharvest storage temperature with exception of β-carotene and rutin. Ripe papaya stored at 25 °C had more carotenoids than those stored at 1 °C. Low (chilling) temperature (1 °C) negatively affected the content of major carotenoids, except β-carotene, but preserved or increased ferulic and caffeic acids levels, as compared to high (safe) temperature (25 °C).


Comparison of radical prostatectomy and iodine 125 interstitial radiotherapy for the treatment of clinically localized prostate cancer: a 7-year biochemical (PSA) progression analysis.
To evaluate the relative efficacy of brachytherapy to radical prostatectomy, we compared biochemical progression rates from a published series of men who underwent iodine 125 (125I) interstitial radiotherapy for localized prostate cancer to a similar group of men who underwent anatomic radical prostatectomy using appropriate end points. Seventy-six men who underwent anatomic radical prostatectomy between 1988 and 1990 were carefully matched for Gleason score and clinical stage to a recently reported contemporary series of patients treated at another institution with 125I brachytherapy without adjuvant treatment. The definition of biochemical progression was a serum PSA level greater than 0.2 ng/mL after anatomic radical prostatectomy and greater than 0.5 ng/mL for brachytherapy-treated patients. The 7-year actuarial PSA progression-free survival following anatomic radical prostatectomy was 97.8% (95% confidence interval [CI], 85.6% to 99.7%) for this group of men selected to match the brachytherapy group, compared to 79% (95% CI not published) for men treated with 125I interstitial radiotherapy. Using comparative end points for biochemical-free progression, failure rates may be higher following 125I interstitial radiotherapy compared to anatomic radical prostatectomy. These data provide a better comparison of biochemical progression than previously published studies and emphasize the need for caution in interpreting the relative efficacy of brachytherapy in controlling localized prostate cancer.


SKL1 Is Essential for Chloroplast Development in Arabidopsis.
The Arabidopsis shikimate kinase-like 1 (skl1-8) mutant is characterized by a pigment-defective phenotype. Although the related phenotypical defect mainly has been attributed to the blocking of chloroplast development, the molecular functions of SKL1 remain largely unknown. In this study, we combined multiple approaches to investigate the potential functions of SKL1. Results showed that the skl1-8 mutant exhibited an albino phenotype and had dramatically reduced chlorophyll content as a consequence of a single nuclear recessive gene mutation. Chemical complementation analysis indicated that SKL1 does not function as SK enzyme in the shikimate pathway. In addition, by chlorophyll fluorescence parameters and immunoblot analysis, the levels of photosynthetic proteins are substantially reduced. Moreover, by transcriptome analysis, specific groups of nuclear genes involved in photosynthesis, such as light-harvesting complex, pigment metabolism, carbon metabolism, and chloroplast gene expression, were down-regulated, whereas several defense and oxidative stress responsive genes were up-regulated in the skl1-8 mutant compared with the wide type. Furthermore, we found the expression of genes related to auxin transport and response was repressed in the skl1-8 mutant, probable suggesting that SKL1 is involved in auxin-related pathways during chloroplast development. Together, these results provide a useful reference for characterization of SKL1 function during chloroplast biogenesis and development.


Mouse CD94 participates in Qa-1-mediated self recognition by NK cells and delivers inhibitory signals independent of Ly-49.
Inhibitory receptors expressed on NK cells recognize MHC class I molecules and transduce negative signals to prevent the lysis of healthy autologous cells. The lectin-like CD94/NKG2 heterodimer has been studied extensively as a human inhibitory receptor. In contrast, in mice, another lectin-like receptor, Ly-49, was the only known inhibitory receptor until the recent discovery of CD94/NKG2 homologues in mice. Here we describe the expression and function of mouse CD94 analyzed by a newly established mAb. CD94 was detected on essentially all NK and NK T cells as well as small fractions of T cells in all mouse strains tested. Two distinct populations were identified among NK and NK T cells, CD94(bright) and CD94(dull) cells, independent of Ly-49 expression. The anti-CD94 mAb completely abrogated the inhibition of target killing mediated by NK recognition of Qa-1/Qdm peptide on target cells. Importantly, CD94(bright) but not CD94(dull) cells were found to be functional in the Qa-1/Qdm-mediated inhibition. In the presence of the mAb, activated NK cells showed substantial cytotoxicity against autologous target cells as well as enhanced cytotoxicity against allogeneic and "missing self" target cells. These results suggest that mouse CD94 participates in the protection of self cells from NK cytotoxicity through the Qa-1 recognition, independent of inhibitory receptors for classical MHC class I such as Ly-49.


Suppression of Type I Interferon Signaling by Flavivirus NS5.
Type I interferon (IFN-I) is the first line of mammalian host defense against viral infection. To counteract this, the flaviviruses, like other viruses, have encoded a variety of antagonists, and use a multi-layered molecular defense strategy to establish their infections. Among the most potent antagonists is non-structural protein 5 (NS5), which has been shown for all disease-causing flaviviruses to target different steps and players of the type I IFN signaling pathway. Here, we summarize the type I IFN antagonist mechanisms used by flaviviruses with a focus on the role of NS5 in regulating one key regulator of type I IFN, signal transducer and activator of transcription 2 (STAT2).


[Morphology and function of the interspinal ligaments and the supraspinal ligament of the lumbar portion of the spine].
The collagenous bundles of fibres in the lumbar interspinous spaces belong to different anatomical structures: the true interspinous ligaments, connecting two neighboured spinous processes in a postero-cranial direction, and fibres, lying in the dorso-caudal part, belonging to the thoracolumbar fascia (aponeurosis of the sacrospinal muscle). In the lumbo-sacral segment there are additional fibres arising from the dorsal part of the 5th lumbar spinous process and descending in a postero-caudal direction, interlacing with the thoracolumbar fascia. Superficial to the thoracolumbar fascia the fibres of the supraspinous ligament pass several spinous processes and form a longitudinal cord. Caudally the supraspinous ligament ends normally at the 4th lumbar spinous process. Below the caudal end of the supraspinous ligament the fibres of the thoracolumbar fascia cross to the opposite side and form a scissor-latticed structure. The interspinous ligaments limit the ventral-flexion of the lumbar spine; they can be extended in extreme retro-flexion, too. Downwards of L4 the fibre-texture of the thoracolumbar fascia permits a greater extent of ventral-flexion. In these segments the erector spinae muscle has to ensure the stability of the spine motion segments instead of a longitudinal ligamentous connection.


Myxofibrosarcoma primary cultures: molecular and pharmacological profile.
Myxofibrosarcoma (MFS), formerly considered as a myxoid variant of malignant fibrous histiocytoma, is the most common sarcoma of the extremities in adults and is characterized by a high frequency of local recurrence. The clinical behavior of MFS is unpredictable and the efficacy of chemotherapy is still not well documented. Furthermore, given the relatively recent recognition of MFS as a distinct pathologic entity its cellular and molecular biology has still not been extensively studied in patient-derived preclinical models. We examined the molecular biology and treatment outcomes of high-grade, patient-derived MFS primary cultures. A total of three patient-derived MFS primary cultures were analyzed. We evaluated the role of CD109 expression and also looked for a correlation between transforming growth factor-beta (TGF-β) expression and sensitivity of the primary cultures to different drugs. CD109 was a promising marker for the identification of more aggressive high-grade MFS and a potential therapeutic target. The results also highlighted the potential role of TGF-β in chemoresistance. Pharmacological analysis confirmed the sensitivity of the cultures to chemotherapy. The most active treatments were epirubicin alone and epirubicin in combination with ifosfamide, the latter representing the current standard of care for soft tissue sarcomas (STSs), including MFS. Our results provide a starting point for further research aimed at improving the management of MFS patients undergoing chemotherapy.


Carboxylesterases expressed in human colon tumor tissue and their role in CPT-11 hydrolysis.
The purpose is to develop new analytical methods to study the expression profile of CPT-11 carboxylesterases and topoisomerase I in colon tumor samples and understand the impact of their expression on CPT-11 metabolism in chemotherapy. We investigated 24 colon tumors for expression of carboxylesterases CES1A1, CES2, CES3, hBr-3, and topoisomerase I genes by real-time PCR and correlated the gene expression with activity assays. The relative abundance of the carboxylesterase isoenzymes and topoisomerase I genes was determined by real-time PCR. Activity assays performed on colon tumor extracts included CPT-11 hydrolase, 4-methylumbelliferyl acetate hydrolase, and topoisomerase I activity assays. Additionally, nondenaturing activity gel electrophoresis with activity staining showed the distribution of carboxylesterases. We detect the expression of CES1A1, CES2, and CES3 carboxylesterase genes in human colon tumors. We were unable to detect the hBr-3 (also called hCE-3) in human liver, colon, or brain. We find large interindividual variation, >/=150-fold, for both CES1A1 and CES3 genes, 23-fold for CES2, and 66-fold for topoisomerase I. Only CES2 gene expression correlated with the carboxylesterase activity assays (P < 0.01) with CPT-11 and 4-methylumbelliferyl acetate as substrates. Nondenaturing activity gel electrophoresis showed that CES2 was the most predominant activity. Topoisomerase I gene expression significantly correlated with topoisomerase I activity (P < 0.01) in the colon tumors, but interindividual variation was very high. We conclude that CES2 is the most abundant carboxylesterase in colon tumors that is responsible for CPT-11 hydrolysis. This pilot study reinforces the hypothesis that there is a large interindividual variation in expression of carboxylesterases that may contribute to variation in therapeutic outcome and/or toxicity of CPT-11 therapy for colon cancer.


Patients' perception of privacy and confidentiality in the emergency department of a busy obstetric unit.
Privacy and confidentiality are central components of patient care and are of particular importance in obstetrics and gynaecology, where clinical situations of a sensitive nature regularly occur. The layout of the emergency department (ED) in maternity units is often not conducive to maintaining privacy. Our study aimed to discover if changing the environment could improve patients' experiences in the ED. We surveyed patients and asked specific questions about their perception of privacy in the ED. We then repeated the survey following renovations to the ED which involved replacing curtained patient areas with walled cubicles. There were 75 pre-renovation surveys and 82 post-renovation surveys completed. Before the renovations took place, only 21% (n = 16) found their privacy to be adequate during their visit to the ED. However this rose to 89% (n = 73) post-renovation. Our study showed that patients' perception of privacy and confidentiality significantly improved following refurbishment of the ED.


[Effect of argon laser photocoagulation on diabetic retinopathy].
To observe the clinical effect of argon laser photocoagulation on diabetic retinopathy (DR) in different stages. A total of 263 eyes in 170 patients with DR in different stages were photocoagulated using argon laser machine 2000. Thirty eyes in non-proliferative stage received focal photocoagulation, 156 pre-proliferative eyes received subtotal panretina photocoagulation, and 77 proliferative eyes were treated with panretinal photocoagulation. Additional focal or grid treatment was given for the eyes with localized or diffused macular edema. The visual acuity, eye fundus examination and retinal fluorescein angiography were carried out 3 months after the treatment and the results compared with the preoperative findings. Improvement of visual acuity by at least one row on the visual chart was achieved in 92 eyes (35%), 144 eyes (55%) exhibited no changes, and 27 eyes (10%) deteriorated. Microaneurysm, retinal edema, exudates and fluorescence leakage were reduced in 89% of the eyes, with the rest eyes requiring additional treatment in the non-proliferative and pre-proliferative stages. For the retina in proliferative stage, neovascularization on the disc resolved completely in 67% eyes, whereas partially in 33% eyes which needed additional photocoagulation. The eyes with partial and complete resolution of macular edema accounted for 91% of the total eyes. Postoperative pre-retinal hemorrhage and vitreous hemorrhage occurred at the rates of 8% and 1%, respectively. Argon laser therapy is effective for the eyes with DR in different stages, but produces better efficacy in non- and pre-proliferative stages than in proliferative stage. Follow-up and additional photocoagulation are necessary for the eyes in proliferative stage to ensure better outcome.


Heart rate and blood pressure response to short-term head-down bed rest: a nonlinear approach.
Although it is well-known that prolonged exposure to microgravity environment such as in space travel results in derangements of orthostasis, recent evidence suggests that even short-term exposure may have similar effects and parallels such common examples as prolonged bed rest. Whereas spectral analysis of heart rate and systolic blood pressure have been unable to detect changes, we hypothesized that nonlinear indexes may be better able to uncover such perturbations. Eighteen healthy subjects were exposed to 4-hour head-down tilt, and of these, 4 exhibited fainting. Two nonlinear indexes, mutual information and recurrence quantification were used to analyze the data. Only recurrence quantification was able to detect a "decoupling" of heart rate and systolic blood pressure at rest using discriminant analysis (p < 0.05). These results suggest that orthostatic intolerance may be due to a decoupling of heart rate from systolic blood pressure reflexive activity occurring at rest.


Cardiac looping may be driven by compressive loads resulting from unequal growth of the heart and pericardial cavity. Observations on a physical simulation model.
The transformation of the straight embryonic heart tube into a helically wound loop is named cardiac looping. Such looping is regarded as an essential process in cardiac morphogenesis since it brings the building blocks of the developing heart into an approximation of their definitive topographical relationships. During the past two decades, a large number of genes have been identified which play important roles in cardiac looping. However, how genetic information is physically translated into the dynamic form changes of the looping heart is still poorly understood. The oldest hypothesis of cardiac looping mechanics attributes the form changes of the heart loop (ventral bending → simple helical coiling → complex helical coiling) to compressive loads resulting from growth differences between the heart and the pericardial cavity. In the present study, we have tested the physical plausibility of this hypothesis, which we call the growth-induced buckling hypothesis, for the first time. Using a physical simulation model, we show that growth-induced buckling of a straight elastic rod within the confined space of a hemispherical cavity can generate the same sequence of form changes as observed in the looping embryonic heart. Our simulation experiments have furthermore shown that, under bilaterally symmetric conditions, growth-induced buckling generates left- and right-handed helices (D-/L-loops) in a 1:1 ratio, while even subtle left- or rightward displacements of the caudal end of the elastic rod at the pre-buckling state are sufficient to direct the buckling process toward the generation of only D- or L-loops, respectively. Our data are discussed with respect to observations made in biological "models." We conclude that compressive loads resulting from unequal growth of the heart and pericardial cavity play important roles in cardiac looping. Asymmetric positioning of the venous heart pole may direct these forces toward a biased generation of D- or L-loops.


A novel breast ultrasound system for providing coronal images: system development and feasibility study.
Breast ultrasound images along coronal plane contain important diagnosis information. However, conventional clinical 2D ultrasound cannot provide such images. In order to solve this problem, we developed a novel ultrasound system aimed at providing breast coronal images. In this system, a spatial sensor was fixed on an ultrasound probe to obtain the image spatial data. A narrow-band rendering method was used to form coronal images based on B-mode images and their corresponding spatial data. Software was developed for data acquisition, processing, rendering and visualization. In phantom experiments, 20 inclusions with different size (5-20 mm) were measured using this new system. The results obtained by the new method well correlated with those measured by a micrometer (y=1.0147x, R(2)=0.9927). The phantom tests also showed that this system had excellent intra- and inter-operator repeatability (ICC>0.995). Three subjects with breast lesions were scanned in vivo using this new system and a commercially available three-dimensional (3D) probe. The average scanning times for the two systems were 64 s and 74 s, respectively. The results revealed that this new method required shorter scanning time. The tumor sizes measured on the coronal plane provided by the new method were smaller by 5.6-11.9% in comparison with the results of the 3D probe. The phantom tests and preliminary subject tests indicated the feasibility of this system for clinical applications by providing additional information for clinical breast ultrasound diagnosis.


Long-term safety of a non-chlorofluorocarbon-containing triamcinolone acetonide inhalation aerosol in patients with asthma. Azmacort HFA Study Group.
In response to environmental concerns regarding chlorofluorocarbon (CFC), two new triamcinolone acetonide (TAA) inhalation aerosol (Azmacort Inhalation Aerosol) formulations have been developed using a more environmentally favorable propellant, HFA-134a (1,1,1,2-tetrafluoroethane). This multicenter, open-label study evaluated the safety of switching asthma patients from TAA-CFC to one of two TAA-HFA formulations. After a 2- or 4-week baseline period during which patients received only CFC-containing TAA Inhaler, 552 patients were randomized to receive TAA-HFA 75 or 225 microg for 6 or 12 months. A total of 493 patients completed treatment. Seven patients discontinued because of adverse events and two because of ineffective asthma control. The incidence of adverse events was similar in the two treatment groups, and most events were mild to moderate in severity and were not considered related to study medication. No clinically relevant suppression of the hypophyseal-pituitary-adrenal (HPA) axis was observed. Pulmonary function tests were not adversely affected by use of either study medication, and improvements were noted in forced expiratory volume in 1 sec (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25%-75%) throughout the course of treatment. This study confirms that TAA-HFA provides effective, long-term asthma control and can safely be substituted for the currently marketed CFC-containing TAA product.


Monte Carlo modeling of light propagation in highly scattering tissues--II: Comparison with measurements in phantoms.
Measurements of the fluence-depth distributions and of the diffuse reflectance of 633 nm light have been made in liquid media with optical properties similar to soft tissues. The results are compared with predictions of Monte Carlo computer calculations in order to test the adequacy of Monte Carlo modeling of light transport in tissue. Except at extremely high albedo, the experimental data and the Monte Carlo results agree well for the depth dependence of the fluence as a function of incident light beam diameter and optical absorption and scattering, and for the dependence of the diffuse reflectance on the albedo. The absolute experimental values for the fluence must be renormalized by a factor which varies with the albedo in order to match the model values, and the possible sources of this discrepancy are discussed.


DEND mutation in Kir6.2 (KCNJ11) reveals a flexible N-terminal region critical for ATP-sensing of the KATP channel.
ATP-sensitive K(+)-channels link metabolism and excitability in neurons, myocytes, and pancreatic islets. Mutations in the pore-forming subunit (Kir6.2; KCNJ11) cause neonatal diabetes, developmental delay, and epilepsy by decreasing sensitivity to ATP inhibition and suppressing electrical activity. Mutations of residue G53 underlie both mild (G53R,S) and severe (G53D) forms of the disease. All examined substitutions (G53D,R,S,A,C,F) reduced ATP-sensitivity, indicating an intolerance of any amino acid other than glycine. Surprisingly, each mutation reduces ATP affinity, rather than intrinsic gating, although structural modeling places G53 at a significant distance from the ATP-binding pocket. We propose that glycine is required in this location for flexibility of the distal N-terminus, and for an induced fit of ATP at the binding site. Consistent with this hypothesis, glycine substitution of the adjacent residue (Q52G) partially rescues ATP affinity of reconstituted Q52G/G53D channels. The results reveal an important feature of the noncanonical ATP-sensing mechanism of K(ATP) channels.


Dispensable role of PTEN in mouse spermatogenesis.
PTEN (phosphatase and tensin homologue deleted on chromosome ten) plays critical roles in multiple cellular processes, including cell proliferation, survival, migration and transformation. A role of PTEN in mammalian spermatogenesis, however, has not been explored. To address this question, we generated a mouse model with PTEN conditional knockout in postnatal male germ cells. We found that spermatogenesis was normal in PTEN-deleted male germ cells. PTEN conditional mutant males produced sperm and sired offspring as competently as wild-type littermates. Moreover, our biochemical analysis also indicated that the Akt (acutely transforming retrovirus AKT8 in rodent T cell lymphoma) signalling pathway was not affected in mutant testis. Taken together, these findings demonstrate that PTEN is dispensable in mouse spermatogenesis.


Nearly monodisperse silica microparticles form in silicone (pre)elastomer mixtures.
The formation of silica from a tetraalkoxysilane in a sol-gel process usually requires a highly polar, typically aqueous, medium that aids in the hydrolysis of the silane and leads to electrostatic stabilization of the growing silica particles. Formation of such silica particles in a hydrophobic medium is much more challenging. We report the formation of silica microspheres within silicone oils (hydroxy-terminated poly(dimethylsiloxane), HO-PDMS) during elastomer cure using atmospheric humidity in a one-pot and one-step synthesis. Using tetraethyl orthosilicate (TEOS) as both cross-linker and silica precursor, and aminopropyl-terminated dimethylsiloxane oligomer (AT-PDMS) as a catalytic surfactant, silica particles of low polydispersity formed near or at the air interface of the elastomer: the presence of a hydrophilic polymer, poly(ethylene glycol) (PEG), had an indirect effect on the particle formation, as it assisted with water transmission into the system, which resulted in particle formation over a wider range of parameters and facilitated silicone elastomer cure further away from the air interface. Depending on the relative humidity during cure, the sizes of particles presenting at the air interface varied from ~6-7 μm under ambient conditions (20-30%RH) to ~7-9 μm at high relative humidity (90% RH). The origin of the controlled particle synthesis is ascribed to the relative solubility of the catalyst and the efficiency of water permeation through the silicone matrix. AT-PDMS preferentially migrates to the air interface, as shown by ninhydrin staining, where it both catalyzes alkoxysilane hydrolysis and condensation, and stabilizes the growing silica particles prior to aggregation. Since reactions in the presence of this catalyst are slow, TEOS can migrate from within the pre-elastomer body to the interface faster than water can penetrate the silicone, such that the main locus of hydrolysis/condensation leading both to silica formation and elastomer cross-linking is at the air interface.


Acute exacerbation of interstitial lung diseases secondary to systemic rheumatic diseases: a prospective study and review of the literature.
Acute exacerbation (AE) is a possible manifestation of interstitial lung diseases (ILD) associated to very high mortality. It's defined as clinically significant respiratory deterioration with evidence of new widespread alveolar abnormalities on computed tomography scan. AE is better described in idiopathic pulmonary fibrosis (IPF) but also reported in ILD secondary to connective tissue diseases (CTD) and vasculitis. The main features and the real clinical impact of this severe complication in these patients are not well defined. Aim of our study was to prospectively investigate the incidence, clinical features and outcome of AE in a population of patients with ILD related to CTD and vasculitis. We consecutively enrolled all patients, with ILD secondary to rheumatic systemic diseases, referring to our multidisciplinary outpatient clinic for rare lung diseases. All patients were followed for at least 12 months (range, 12-36 months). At baseline, all patients underwent to a core set of laboratory investigations and periodically followed; data about demographic, disease onset, clinical, serological and therapeutic features were also recorded. AE occurred in 9/78 patients, with an incidence of 5.77/100 patients/year, and 5/9 patients died because of AE. The baseline value of DLCO was significantly associated to the risk of AE at Cox regression. In patients with ILD related to rheumatic systemic diseases AE can occur with an incidence similar to IPF. Rheumatologists should carefully consider this life-threatening complication as a possible natural course of all patients with ILD secondary to systemic rheumatic disease.


Effects of sports participation on psychiatric symptoms and brain activations during sports observation in schizophrenia.
Weight gain has been identified as being responsible for increased morbidity and mortality rates of schizophrenia patients. For the management of weight gain, exercise is one of the most acknowledged interventions. At the same time, exercise and sports have been recognized for their positive impact on psychiatric symptoms of schizophrenia. However, the neurobiological basis for this remains poorly understood. We aimed to examine the effect of sports participation on weight gain, psychiatric symptoms and brain activation during sports observation in schizophrenia patients. Thirteen schizophrenia patients who participated in a 3-month program, including sports participation and 10 control schizophrenia patients were studied. In both groups, body mass index (BMI), Positive and Negative Syndrome Scale (PANSS), and brain activation during observation of sports-related actions measured by functional magnetic resonance imaging were accessed before and after a 3-month interval. BMI and general psychopathology scale of PANSS were significantly reduced in the program group but not in the control group after a 3-month interval. Compared with baseline, activation of the body-selective extrastriate body area (EBA) in the posterior temporal-occipital cortex during observation of sports-related actions was increased in the program group. In this group, increase in EBA activation was associated with improvement in the general psychopathology scale of PANSS. Sports participation had a positive effect not only on weight gain but also on psychiatric symptoms in schizophrenia. EBA might mediate these beneficial effects of sports participation. Our findings merit further investigation of neurobiological mechanisms underlying the therapeutic effect of sports for schizophrenia.


Real-time face detection and lip feature extraction using field-programmable gate arrays.
This paper proposes a new technique for face detection and lip feature extraction. A real-time field-programmable gate array (FPGA) implementation of the two proposed techniques is also presented. Face detection is based on a naive Bayes classifier that classifies an edge-extracted representation of an image. Using edge representation significantly reduces the model's size to only 5184 B, which is 2417 times smaller than a comparable statistical modeling technique, while achieving an 86.6% correct detection rate under various lighting conditions. Lip feature extraction uses the contrast around the lip contour to extract the height and width of the mouth, metrics that are useful for speech filtering. The proposed FPGA system occupies only 15050 logic cells, or about six times less than a current comparable FPGA face detection system.


Specific chromatin landscapes and transcription factors couple breast cancer subtype with metastatic relapse to lung or brain.
Few somatic mutations have been linked to breast cancer metastasis, whereas transcriptomic differences among primary tumors correlate with incidence of metastasis, especially to the lungs and brain. However, the epigenomic alterations and transcription factors (TFs) which underlie these alterations remain unclear. To identify these, we performed RNA-seq, Chromatin Immunoprecipitation and sequencing (ChIP-seq) and Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) of the MDA-MB-231 cell line and its brain (BrM2) and lung (LM2) metastatic sub-populations. We incorporated ATAC-seq data from TCGA to assess metastatic open chromatin signatures, and gene expression data from human metastatic datasets to nominate transcription factor biomarkers. Our integrated epigenomic analyses found that lung and brain metastatic cells exhibit both shared and distinctive signatures of active chromatin. Notably, metastatic sub-populations exhibit increased activation of both promoters and enhancers. We also integrated these data with chromosome conformation capture coupled with ChIP-seq (HiChIP) derived enhancer-promoter interactions to predict enhancer-controlled pathway alterations. We found that enhancer changes are associated with endothelial cell migration in LM2, and negative regulation of epithelial cell proliferation in BrM2. Promoter changes are associated with vasculature development in LM2 and homophilic cell adhesion in BrM2. Using ATAC-seq, we identified a metastasis open-chromatin signature that is elevated in basal-like and HER2-enriched breast cancer subtypes and associates with worse prognosis in human samples. We further uncovered TFs associated with the open chromatin landscapes of metastatic cells and whose expression correlates with risk for metastasis. While some of these TFs are associated with primary breast tumor subtypes, others more specifically correlate with lung or brain metastasis. We identify distinctive epigenomic properties of breast cancer cells that metastasize to the lung and brain. We also demonstrate that signatures of active chromatin sites are partially linked to human breast cancer subtypes with poor prognosis, and that specific TFs can independently distinguish lung and brain relapse.


An In Vivo CRISPR Screen Identifies Stepwise Genetic Dependencies of Metastatic Progression.
Blood-borne metastasis of breast cancer involves a series of tightly regulated sequential steps, including the growth of a primary tumor lesion, intravasation of circulating tumor cells (CTC), and adaptation in various distant metastatic sites. The genes orchestrating each of these steps are poorly understood in physiologically relevant contexts, owing to the rarity of experimental models that faithfully recapitulate the biology, growth kinetics, and tropism of human breast cancer. Here, we conducted an in vivo loss-of-function CRISPR screen in newly derived CTC xenografts, unique in their ability to spontaneously mirror the human disease, and identified specific genetic dependencies for each step of the metastatic process. Validation experiments revealed sensitivities to inhibitors that are already available, such as PLK1 inhibitors, to prevent CTC intravasation. Together, these findings present a new tool to reclassify driver genes involved in the spread of human cancer, providing insights into the biology of metastasis and paving the way to test targeted treatment approaches. A loss-of-function CRISPR screen in human CTC-derived xenografts identifies genes critical for individual steps of the metastatic cascade, suggesting novel drivers and treatment opportunities for metastatic breast cancers.


Sinonasal cancer and occupational exposure in a population-based registry.
We examined occupational exposures among subjects with sinonasal cancer (SNC) recorded in a population-based registry in the Lombardy Region, the most populated and industrialized Italian region. The registry collects complete clinical information and exposure to carcinogens regarding all SNC cases occurring in the population of the region. In the period 2008-2011, we recorded 210 SNC cases (137 men, 73 women). The most frequent occupational exposures were to wood (44 cases, 21.0%) and leather dust (29 cases, 13.8%), especially among men: 39 cases (28.5%) to wood and 23 cases (16.8%) to leather dust. Exposure to other agents was infrequent (<2%). Among 62 subjects with adenocarcinoma, 50% had been exposed to wood dust and 30.7% to leather dust. The proportions were around 10% in subjects with squamous cell carcinoma and about 20% for tumors with another histology. The age-standardized rates (×100,000 person-years) were 0.7 in men and 0.3 in women. Complete collection of cases and their occupational history through a specialized cancer registry is fundamental to accurately monitor SNC occurrence in a population and to uncover exposure to carcinogens in different industrial sectors, even those not considered as posing a high risk of SNC, and also in extraoccupational settings.


Fabrication of multifunctional ferric oxide nanoparticles for tumor-targeted magnetic resonance imaging and precise photothermal therapy with magnetic field enhancement.
In this study, a biocompatible nanotheranostic platform (termed as Fe2O3@PDA-affibody) has been constructed on the basis of coating a near-infrared light (NIR)-absorbing polydopamine (PDA) shell on oleic acid-capped superparamagnetic ferric oxide nanoparticles (Fe2O3 NPs) using the water-in-oil microemulsion method and then conjugated with affibody ZIGF1R:4551, a peptide with high affinity to tumor and a polyethylene glycol (PEG) stabilizer. The Fe2O3@PDA-affibody integrates T2-weighted magnetic resonance imaging (MRI), tumor-targeting, and magnetic field (MF)-enhanced photothermal therapy (PTT) functionalities into an all-in-one system. The Fe2O3@PDA-affibody shows high negative contrast in the MRI of an SW620 tumor bearing mouse with a decrease of 68% MRI signal, indicating that the Fe2O3@PDA-affibody can recognize tumor with high efficacy and specificity. Furthermore, a high accumulation ratio (>13.5% ID g-1) and enhanced inhibition of tumor growth are achieved under near-infrared (NIR) (808 nm) laser irradiation with the aid of an external MF focused on the targeted tumor, resulting in complete eradication of mouse-borne SW620 tumors without regrowth.


Determination of phthalates in bottled water by automated on-line solid phase extraction coupled to liquid chromatography with uv detection.
An on-line solid phase extraction coupled to liquid chromatography with UV detection (SPE/LC-UV) method was automated by the multisyringe flow-injection analysis (MSFIA) system for the determination of three phthalic acid esters (PAEs). The PAEs determined in drinking water stored in polyethylene terephthalate (PET) bottles of ten commercial brands were dimethyl phthalate (DMP), diethyl phthalate (DEP) and dibutyl phthalate (DBP). C18-bonded silica membrane was used for isolation and enrichment of the PAEs in water samples. The calibration range of the SPE/LC-UV method was 2.5-100μgL-1 for DMP and DEP and 10-100μgL-1 for DBP with correlation coefficients (r) ranging from 0.9970 to 0.9975. Limits of detection (LODs) were between 0.7 and 2.4μgL-1. Inter-day reproducibility performed at two concentration levels (10 and 100μgL-1) expressed as relative standard deviation (%RSD) were found in the range of 0.9-4.0%. The solvent volume was reduced to 18mL with a total analysis time of 48min per sample. The major species detected in bottled water samples was DBP reaching concentrations between 20.5 and 82.8μgL-1. The recovery percentages for the three analytes in drinking water were 80-115%. The migration test showed a great variation in the sum of migrated PAEs level (10.2-50.6μgL-1) among the PET bottle brands analyzed indicating that the presence of these contaminants in the plastic containers may depend on raw materials and the conditions used during their production process.


Crystal structures of the Toxoplasma gondii hypoxanthine-guanine phosphoribosyltransferase-GMP and -IMP complexes: comparison of purine binding interactions with the XMP complex.
The crystal structures of the guanosine 5'-monophosphate (GMP) and inosine 5'-monophosphate (IMP) complexes of Toxoplasma gondii hypoxanthine-guanine phosphoribosyltransferase (HGPRT) have been determined at 1.65 and 1.90 A resolution. These complexes, which crystallize in space groups P2(1) (a = 65.45 A, b = 90.84 A, c = 80. 26 A, and beta = 92.53 degrees ) and P2(1)2(1)2(1) (a = 84.54 A, b = 102.44 A, and c = 108.83 A), each comprise a tetramer in the crystallographic asymmetric unit. All active sites in the tetramers are fully occupied by the nucleotide. Comparison of these structures with that of the xanthosine 5'-monophosphate (XMP)-pyrophosphate-Mg(2+) ternary complex reported in the following article [Héroux, A., et al. (1999) Biochemistry 38, 14495-14506] shows how T. gondii HGPRT is able to recognize guanine, hypoxanthine, and xanthine as substrates, and suggests why the human enzyme cannot use xanthine efficiently. Comparison with the apoenzyme reveals the structural changes that occur upon binding of purines and ribose 5'-phosphate to HGPRT. Two structural features important to the HGPRT mechanism, a previously unrecognized active site loop (loop III', residues 180-184) and an active site peptide bond (Leu78-Lys79) that adopts both the cis and the trans configurations, are presented.


Comparison of five in vitro digestion models to in vivo experimental results: lead bioaccessibility in the human gastrointestinal tract.
This paper presents a multi-laboratory comparison study of in vitro models assessing bioaccessibility of soil-bound lead in the human gastrointestinal tract under simulated fasted and fed conditions. Oral bioavailability data from a previous human in vivo study on the same soil served as a reference point. In general, the bioaccessible lead fraction was significantly (P<0.05) different between the in vitro methods and ranged for the fasted models from 2% to 33% and for the fed models from 7% to 29%. The in vivo bioavailability data from literature were 26.2+/-8.1% for fasted conditions, compared to 2.5+/-1.7% for fed conditions. Under fed conditions, all models returned higher bioaccessibility values than the in vivo bioavailability; whereas three models returned a lower bioaccessibility than bioavailability under fasted conditions. These differences are often due to the method's digestion parameters that need further optimization. An important outcome of this study was the determination that the method for separating the bioaccessible lead from the non-bioaccessible fraction (centrifugation, filtration, ultrafiltration) is crucial for the interpretation of the results. Bioaccessibility values from models that use more stringent separation methods better approximate in vivo bioavailability results, yet at the expense of the level of conservancy. We conclude from this study that more optimization of in vitro digestion models is needed for use in risk assessment. Moreover, attention should be paid to the laboratory separation method since it largely influences what fraction of the contaminant is considered bioaccessible.


Perceived distributed effort in team ball sports.
In this study, we explored the multifaceted concept of perceived mental and physical effort in team sport contexts where athletes must invest individual and shared efforts to reach a common goal. Semi-structured interviews were conducted with a convenience sample of 15 Catalan professional coaches (3 women and 12 men, 3 each from the following sports: volleyball, basketball, handball, soccer, and water polo) to gain their views of three perceived effort-related dimensions: physical, psychological, and tactical. From a theoretical thematic analysis, it was found that the perception of effort is closely related to how effort is distributed within the team. Moreover, coaches viewed physical effort in relation to the frequency and intensity of the players' involvement in the game. They identified psychological effort in situations where players pay attention to proper cues, and manage emotions under difficult circumstances. Tactical effort addressed the decision-making process of players and how they fulfilled their roles while taking into account the actions of their teammates and opponents. Based on these findings, a model of perceived distributed effort was developed, which delineates the elements that compose each of the aforementioned dimensions. Implications of perceived distributed effort in team coordination and shared mental models are discussed.


Interference of retroviral envelope with vaccine-induced CD8+ T cell responses is relieved by co-administration of cytokine-encoding vectors.
Retroviral envelope (Env) proteins are known to exhibit immunosuppressive properties, which become apparent not only in retroviral infections, but also in gene-based immunizations using retroviral immunogens, where envelope interferes with the induction of CD8+ T cell responses towards another, simultaneously or subsequently delivered, immunogen. In the Friend retrovirus mouse model, immunization with a plasmid encoding the Friend murine leukemia virus (F-MuLV) Leader-Gag protein resulted in induction of a strong GagL85-93-specific CD8+ T cell response, while the response was completely abrogated by co-immunization with an F-MuLV Env-encoding plasmid. In order to overcome this interference of retroviral envelope, we employed plasmids encoding the cytokines interleukin (IL) 1β, IL2, IL12, IL15, IL21, IL28A or granulocyte-macrophage colony-stimulating factor (GM-CSF) as genetic adjuvants. Co-application of plasmids encoding IL2, IL12, IL21, IL28A and especially GM-CSF rescued the induction of GagL85-93-specific CD8+ T cells in mice vaccinated with FV Leader-Gag and Env. Mice that were immunized with plasmids encoding Leader-Gag and Env and the cytokines IL1β, IL12, IL15, IL28A or GM-CSF, but not Leader-Gag and Env without any cytokine, showed significantly reduced viral loads upon a high-dose Friend virus challenge infection. Our data demonstrate the potency of cytokine-encoding vectors as adjuvants and immune modulators in composite vaccines for anti-retroviral immunization.


Open Randomized Clinical Trial on JWSJZ Decoction for the Treatment of ALS Patients.
Objective. To investigate the efficacy and safety of the traditional Chinese medicine Jiawei Sijunzi (JWSJZ) decoction for the treatment of patients with amyotrophic lateral sclerosis (ALS). Methods. Forty-eight patients with ALS were divided into a JWSJZ group (n = 24) and a control group (n = 24) using a randomized number method. Together with the basic treatment for ALS, JWSJZ decoction was added to the treatment regimen of patients in the JWSJZ group or Riluzole was administered to the control group for 6 months. Neurologists evaluated the treated and control patients using the ALS functional rating scale (ALSFRS) before, 3 and 6 months after starting the additional treatments. Results. The ALSFRS scores in both groups were lower 3 and 6 months after treatment than before. There was a significant difference at 6 months after treatment between the subgroups of patients with ALS whose limbs were the initial site of attack. No serious adverse effects were observed in the JWSJZ group. Conclusion. JWSJZ decoction may be a safe treatment for ALS, and may have delayed the development of ALS, especially in the subgroup of patients in whom the limbs were attacked first when compared with Riluzole treatment.


Evaluation of a spectrophotometric method for canine serum lipase determination.
The British antilewisite butyrate-dithionitrobenzoate (BALB-DTNB) spectrophotometric serum lipase assay was evaluated for precision, accuracy, and diagnostic usefulness in analyzing canine sera. Sera samples from clinically healthy dogs, dogs with experimentally induced pancreatitis, and dogs with spontaneous pancreatitis were analyzed. A titrimetric method of serum lipase determination was used for comparison. Although the BALB-DTNB method was not found to be precise or accurate for determining the lipase activity of canine serum samples, it seemed to be at least as diagnostically useful as the titrimetric procedure. The small sample size requirement and the speed of analysis of the BALB-DTNB procedure are advantages of this method over the titrimetric method, and thus, its use in place of the titrimetric method is justified. A laboratory reference range of 3 to 37 IU/L was determined for canine serum.


Use of a Bayesian model including QTL markers increases prediction reliability when test animals are distant from the reference population.
Relatedness between reference and test animals has an important effect on the reliability of genomic prediction for test animals. Because genomic prediction has been widely applied in practical cattle breeding and bulls have been selected according to genomic breeding value without progeny testing, the sires or grandsires of candidates might not have phenotypic information and might not be in the reference population when the candidates are selected. The objective of this study was to investigate the decreasing trend of the reliability of genomic prediction given distant reference populations, using genomic best linear unbiased prediction (GBLUP) and Bayesian variable selection models with or without including the quantitative trait locus (QTL) markers detected from sequencing data. The data used in this study consisted of 22,242 bulls genotyped using the 54K SNP array from EuroGenomics. Among them, 1,444 Danish bulls born from 2006 to 2010 were selected as test animals. Different reference populations with varying relationships to test animals were created according to pedigree-based relationships. The reference individuals having a relationship with one or more test animals higher than 0.4 (scenario ρ < 0.4), 0.2 (ρ < 0.2), or 0.1 (ρ < 0.1, where ρ = relationship coefficient) were removed from reference sets; these represented the distance between reference and test animals being 2 generations, 3 generations, and 4 generations, respectively. Imputed whole-genome sequencing data of bulls from Denmark were used to conduct a genome-wide association study (GWAS). A small number of significant variants (QTL markers) from the GWAS were added to the array data. To compare the effects of different models, the basic GBLUP model, a Bayesian selection variable model, a GBLUP model with 2 components of genetic effects, and a Bayesian model with pooled array data and QTL markers were used for estimating genomic estimated breeding values (GEBV) of test animals. The reliability of genomic prediction decreased when the test animals were more generations away from the reference population. The reliability of genomic prediction was 0.461 for 1 generation away and 0.396 for 3 generations away, with the same number of individuals in the reference set, using a GBLUP model with chip markers only. The results showed that using the Bayesian method and QTL markers improved the reliability of genomic prediction in all scenarios of relationship between test and reference animals, in a range of 1.3% and 65.1% (4 generations away with only 841 individuals in the reference set). However, most gains were for predictions of milk yield and fat yield. There was little improvement for predictions of protein yield and mastitis, and no improvement for prediction of fertility, except for scenario ρ < 0.1, in which there was a large improvement for predictions of all traits. On the other hand, models including more than 10% polygenic effect decreased prediction reliability when the relationship between test and reference animals was distant.


Natural variation of piRNA expression affects immunity to transposable elements.
In the Drosophila germline, transposable elements (TEs) are silenced by PIWI-interacting RNA (piRNA) that originate from distinct genomic regions termed piRNA clusters and are processed by PIWI-subfamily Argonaute proteins. Here, we explore the variation in the ability to restrain an alien TE in different Drosophila strains. The I-element is a retrotransposon involved in the phenomenon of I-R hybrid dysgenesis in Drosophila melanogaster. Genomes of R strains do not contain active I-elements, but harbour remnants of ancestral I-related elements. The permissivity to I-element activity of R females, called reactivity, varies considerably in natural R populations, indicating the existence of a strong natural polymorphism in defense systems targeting transposons. To reveal the nature of such polymorphisms, we compared ovarian small RNAs between R strains with low and high reactivity and show that reactivity negatively correlates with the ancestral I-element-specific piRNA content. Analysis of piRNA clusters containing remnants of I-elements shows increased expression of the piRNA precursors and enrichment by the Heterochromatin Protein 1 homolog, Rhino, in weak R strains, which is in accordance with stronger piRNA expression by these regions. To explore the nature of the differences in piRNA production, we focused on two R strains, weak and strong, and showed that the efficiency of maternal inheritance of piRNAs as well as the I-element copy number are very similar in both strains. At the same time, germline and somatic uni-strand piRNA clusters generate more piRNAs in strains with low reactivity, suggesting the relationship between the efficiency of primary piRNA production and variable response to TE invasions. The strength of adaptive genome defense is likely driven by naturally occurring polymorphisms in the rapidly evolving piRNA pathway proteins. We hypothesize that hyper-efficient piRNA production is contributing to elimination of a telomeric retrotransposon HeT-A, which we have observed in one particular transposon-resistant R strain.


[The activity of peroxidation of lipids in biologic samples of patients with recurrent diseases of upper respiratory tracts].
The parameters of peroxidation of lipids in blood and saliva were analyzed on sampling of 60 patients with recurrent diseases of upper respiratory tracts during period of remission. It is established that the content of primary and secondary products of peroxidation of neutral lipids in blood plasma is lower as compared with healthy persons. The content of end products of peroxidation of neutral lipids and products of oxidation of phospholipids in blood plasma is higher as compared with healthy persons. The characteristics of alterations of intermediates of peroxidation of lipids in various biologic samples depend on the localization of inflammatory process.


Effect of temperature shifts on extracellular proteinase-specific mRNA pools in Pseudomonas fluorescens B52.
The influence of a shift in temperature from 20 to 32 degrees C on extracellular proteinase synthesis by Pseudomonas fluorescens B52 was examined. When cells actively synthesizing proteinase at 20 degrees C were shifted to 32 degrees C, enzyme synthesis ceased immediately. After 30 min at 32 degrees C, cells recovered at 20 degrees C after a lag of 30 min. Rifampin and chloramphenicol prevented recovery of synthesis at 20 degrees C. Rifampin-insensitive proteinase synthesis (an indirect measure of proteinase-specific mRNA pools) decreased after the exposure of cells to 32 degrees C for 30 min but was recovered during incubation at 20 degrees C. Controls not exposed to a temperature shift experienced no loss of rifampin-independent synthesis. Cells experienced a 50% reduction in mRNA pools after 15 min at 32 degrees C. The data support the working hypothesis that the loss of mRNA pools after treatment at 32 degrees C is responsible for the lag before the recovery of extracellular proteinase synthesis.


[The analysis of dosage inaccuracy and its minimization modes].
The article deals with the analysis of factors impacting dosage inaccuracy and input of this inaccuracy into total inaccuracy of analysis. The total dosage inaccuracy includes both the value of inaccuracy mentioned in the technical passport of dosage device and inaccuracy values occurring at all stages of dosage during implementation of technique. The characteristics of dosage devices and their merits and shortcomings are taken into account. The article substantiates the choice of types of dosage devices to be implemented in the laboratory research. The modes of errors elimination and inaccuracy minimization were analyzed against the perfectly calibrated and totally corresponded to passport data device of air transfer dosage.


The association between adolescent football participation and early adulthood depression.
Concerned about potentially increased risk of neurodegenerative disease, several health professionals and policy makers have proposed limiting or banning youth participation in American-style tackle football. Given the large affected population (over 1 million boys play high school football annually), careful estimation of the long-term health effects of playing football is necessary for developing effective public health policy. Unfortunately, existing attempts to estimate these effects tend not to generalize to current participants because they either studied a much older cohort or, more seriously, failed to account for potential confounding. We leverage data from a nationally representative cohort of American men who were in grades 7-12 in the 1994-95 school year to estimate the effect of playing football in adolescent on depression in early adulthood. We control for several potential confounders related to subjects' health, behavior, educational experience, family background, and family health history through matching and regression adjustment. We found no evidence of even a small harmful effect of football participation on scores on a version of the Center for Epidemiological Studies Depression scale (CES-D) nor did we find evidence of adverse associations with several secondary outcomes including anxiety disorder diagnosis or alcohol dependence in early adulthood. For men who were in grades 7-12 in the 1994-95 school year, participating or intending to participate in school football does not appear to be a major risk factor for early adulthood depression.


SLC26 Anion Transporters.
Solute carrier family 26 (SLC26) is a family of functionally diverse anion transporters found in all kingdoms of life. Anions transported by SLC26 proteins include chloride, bicarbonate, and sulfate, but also small organic dicarboxylates such as fumarate and oxalate. The human genome encodes ten functional homologs, several of which are causally associated with severe human diseases, highlighting their physiological importance. Here, we review novel insights into the structure and function of SLC26 proteins and summarize the physiological relevance of human members.


Stochastic multitype epidemics in a community of households: estimation and form of optimal vaccination schemes.
This paper treats a stochastic model for an SIR (susceptible-->infective-->removed) multitype household epidemic. The community is assumed to be closed, individuals are of different types and each individual belongs to a household. Previously obtained probabilistic and inferential results for the model are used to derive the optimal vaccination scheme. By this is meant the scheme that vaccinates the fewest among all vaccination schemes that reduce the threshold parameter below 1. This is done for the situation where all model parameters are known and also for the case where parameters are estimated from an outbreak in the community prior to vaccination. It is shown that the algorithm which chooses vaccines sequentially, at each step selecting the individual which reduces the threshold parameter the most, is not in general an optimal scheme. As a consequence, explicit characterisation of the optimal scheme is only possible in certain special cases. Two different types of vaccine responses, leaky and all-or-nothing, are considered and compared for the problems mentioned above. The methods are illustrated with some numerical examples.


RGD-Modified Titanium as an Improved Osteoinductive Biomaterial for Use in Dental and Orthopedic Implants.
This study describes the synthesis, surface analysis, and biological evaluation of bioactive titanium surfaces. The aim was to achieve an improved effect on osteoinduction in dental and orthopedic implants. For this purpose, a chemistry was developed, which allows to bind the bioactive cyclopeptide cRGDfK covalently to biomedically used titanium via polyethylene glycol linkers of different lengths. The chemical process is practicable, robust, and metal-free. The resulting chemically modified titanium plates show improved osteoinductive properties. The modification with cRGDfK targets the integrin αvβ3, which is highly expressed in osteoblasts and is essential for many basic functions in the development of bone tissue. The successful immobilization of cRGDfK on titanium surfaces has been demonstrated by contact angle measurements and X-ray photoelectron spectroscopy. We show in in vitro studies that the presence of the cRGDfK peptide on titanium surfaces has a positive effect on bone formation.


Concussion in University Level Sport: Knowledge and Awareness of Athletes and Coaches.
Using a cross-sectional survey concussion knowledge was evaluated among forty university-level athletes (n = 20, rugby union players; n = 20, Gaelic football players) and eight experienced team coaches (n = 2, rugby union; n = 2, Gaelic football; n = 1, soccer; n = 1, hockey; n = 1, netball; n = 1, basketball). Levels of knowledge of concussion were high across all participants. Coaches had higher knowledge scores for almost all areas; however, there was evidence of important gaps even in this group. Knowledge was not sufficient in identifying concussion, and when it is safe to return to play following a concussion. Impaired knowledge of how to recognise a concussion, and misunderstanding the need for rest and rehabilitation before return to play presents a hazard to health from second impact and more catastrophic brain injury. We discuss reasons for these guideline misconceptions, and suggest that attitude issues on the significance of concussion may underlie a willingness to want to play with a concussion. This suggests the current education on sport-related concussion needs to be expanded for the appropriate management of university-level contact sports.


Human Endometrial Fibroblasts Derived from Mesenchymal Progenitors Inherit Progesterone Resistance and Acquire an Inflammatory Phenotype in the Endometrial Niche in Endometriosis.
Human endometrium undergoes cyclic regeneration involving stem/progenitor cells, but the role of resident endometrial mesenchymal stem cells (eMSC) as progenitors of endometrial stromal fibroblasts (eSF) has not been definitively demonstrated. In endometriosis, eSF display progesterone (P4) resistance with impaired decidualization in vivo and in vitro. To investigate eMSC as precursors of eSF and whether endometriosis P4 resistance is inherited from eMSC, we analyzed transcriptomes of eutopic endometrium eMSC and eSF isolated by fluorescence-activated cell sorting (FACS) from endometriosis (eMSCendo, eSFendo) and controls (eMSCcontrol, eSFcontrol) and their derived primary cultures. Differentially expressed lineage-associated genes (LG) of FACS-isolated eMSC and eSF were largely conserved in endometriosis. In culture, eSFcontrol maintained in vitro expression of a subset of eSF LG and decidualized in vitro with P4 The eMSCcontrol cultures differentiated in vitro to eSF lineage, down-regulating eMSC LG and up-regulating eSF LG, showing minimal transcriptome differences versus eSFcontrol cultures and decidualizing in vitro. Cultured eSFendo displayed less in vitro LG stability and did not decidualize in vitro. In vitro, eMSCendo differentiated to eSF lineage but showed more differentially expressed genes versus eSFendo cultures, and did not decidualize in vitro, demonstrating P4 resistance inherited from eMSCendo Compared to controls, cultures from tissue-derived eSFendo uniquely had a pro-inflammatory phenotype not present in eMSCendo differentiated to eSF in vitro, suggesting divergent niche effects for in vivo versus in vitro lineage differentiation. These findings substantiate eMSC as progenitors of eSF and reveal eSF in endometriosis as having P4 resistance inherited from eMSC and a pro-inflammatory phenotype acquired within the endometrial niche.


Integrating chemical and genetic silencing strategies to identify host kinase-phosphatase inhibitor networks that control bacterial infection.
Every year three million people die as a result of bacterial infections, and this number may further increase due to resistance to current antibiotics. These antibiotics target almost all essential bacterial processes, leaving only a few new targets for manipulation. The host proteome has many more potential targets for manipulation in order to control bacterial infection, as exemplified by the observation that inhibiting the host kinase Akt supports the elimination of different intracellular bacteria including Salmonella and M. tuberculosis. If host kinases are involved in the control of bacterial infections, phosphatases could be as well. Here we present an integrated small interference RNA and small molecule screen to identify host phosphatase-inhibitor combinations that control bacterial infection. We define host phosphatases inhibiting intracellular growth of Salmonella and identify corresponding inhibitors for the dual specificity phosphatases DUSP11 and 27. Pathway analysis places many kinases and phosphatases controlling bacterial infection in an integrated pathway centered around Akt. This network controls host cell metabolism, survival, and growth and bacterial survival and reflect a natural host cell response to bacterial infection. Inhibiting two enzyme classes with opposite activities-kinases and phosphatases-may be a new strategy to overcome infections by antibiotic-resistant bacteria.