Hospital Prices Grew Substantially Faster Than Physician Prices For Hospital-Based Care In 2007-14.
Evidence suggests that growth in providers' prices drives growth in health care spending on the privately insured. However, existing work has not systematically differentiated between the growth rate of hospital prices and that of physician prices. We analyzed growth in both types of prices for inpatient and hospital-based outpatient services using actual negotiated prices paid by insurers. We found that in the period 2007-14 hospital prices grew substantially faster than physician prices. For inpatient care, hospital prices grew 42 percent, while physician prices grew 18 percent. Similarly, for hospital-based outpatient care, hospital prices grew 25 percent, while physician prices grew 6 percent. A majority of the growth in payments for inpatient and hospital-based outpatient care was driven by growth in hospital prices, not physician prices. Our work suggests that efforts to reduce health care spending should be primarily focused on addressing growth in hospital rather than physician prices. Policy makers should consider a range of options to address hospital price growth, including antitrust enforcement, administered pricing, the use of reference pricing, and incentivizing referring physicians to make more cost-efficient referrals.


Distinct patterns of infiltrating CD8+ T cells in HPV+ and CD68 macrophages in HPV- oropharyngeal squamous cell carcinomas are associated with better clinical outcome but PD-L1 expression is not prognostic.
Immunotherapies are beginning to revolutionise treatment paradigms in oncology with monoclonal antibodies (mAb) targeting T-cell co-inhibitory (e.g. PD-1/PD-L1) and co-stimulatory pathways (e.g. CTLA-4/CD28) demonstrating clinical utility. Some clinical studies demonstrate that responsiveness to PD-1/PD-L1 mAb therapy is greater in patients with expression of PD-L1 in the tumour microenvironment. However, robust responses have also been observed in patients with low or absent expression of PD-L1. Using multiplex immuno-fluorescent labelling we sought to determine how infiltration of tumours by CD8+ T-cells, their expression of PD-1, and the expression of PD-L1 on both tumours and CD68 cells (macrophages) correlated with HPV status and outcome in a cohort of 124 oropharyngeal squamous cell carcinomas (OPSCC).


Feasibility of Dental Implant Replacement in Failed Sites: A Systematic Review.
To assess the clinical outcomes of replaced implants after removal of failed ones. In addition, associated risk factors that might affect the final outcome of these procedures were also explored. An electronic literature search was conducted by two reviewers in several databases for articles written in English up to November 2014. Human clinical trials with a minimum of 10 subjects enrolled that reported clinical outcomes with a mean follow-up period of at least 12 months after implant replacement were included. Implant survival and nonmodifiable/modifiable factors at second and third implant placement attempts were studied. Hence, the PICO question that was aimed to be addressed was: Do patients undergoing implant replacement (second and third attempts) in previous failed sites have comparable clinical outcomes by means of implant survival/failure rate to implants placed at the first attempt? Five retrospective clinical cohort studies and two case series satisfied the selection criteria and thus were included in this review. In total, 396 patients were studied due to implant replacement in previous failed sites. The survival rate for implant replacement at the second attempt was 88.84% (390/439; range, 71% to 94.6%) with a mean follow-up of 41.59 ± 16.77 months. Thirty-one implants were replaced for a third attempt with a mean survival rate of 74.19% (23/31) at the follow-up of 29.66 ± 14.71 months. Major risk indicators were generally divided into patient-related factors (health status, smoking habits, and oral hygiene maintenance), implant characteristics (dimensions, coating, and loading), and site characteristics (bone quality and density, vertical and horizontal dimensions, soft tissue around the implant). Implant replacement is a reasonably feasible option for scenarios of early and late implant failure. However, modifiable risk factors must be controlled before proceeding for implant replacement.


Mycobacterial growth and bacterial contamination in the mycobacteria growth indicator tube and BACTEC 460 culture systems.
The BACTEC 460 system currently provides the most rapid detection of mycobacterial growth, but the system is radiometric and requires needles to inoculate specimens through the bottle's septum. The Mycobacteria Growth Indicator Tube (MGIT) system has a liquid medium, like the BACTEC system, and does not require needles when inoculating specimens. We compared mycobacterial growth from 510 specimens in the two systems. Average time to acid-fast bacillus (AFB) detection and identification to the species level was less with the BACTEC system, but this result was statistically significant only for AFB detection in specimens containing Mycobacterium avium-M. intracellulare complex. The contamination rate with MGIT was 29%; the BACTEC rate was 5%. To investigate MGIT contamination, we initiated a second study with changes in specimen processing. The MGIT contamination rate was reduced to 12%; the BACTEC rate was not significantly affected (5.5%). The most likely explanation for the contamination in MGIT is the richness of its medium compared to the BACTEC medium. Cost analysis for the two systems in a laboratory that processes 4,500 specimens a year is presented. The data suggest that the BACTEC 460 and the MGIT systems are approximately equivalent in cost and ability to support the growth of AFB. The MGIT system appears safer and easier to use and was preferred by laboratory personnel, but it cannot currently be used for blood specimens or antituberculosis susceptibility testing.


Clinical feasibility of Axillary Reverse Mapping and its influence on breast cancer related lymphedema: a systematic review.
Breast cancer is the most common malignancy in women worldwide. Fortunately, the overall survival is good. Therefore it is important to focus on the morbidities related to breast cancer treatment. One of the most dreaded morbidities is lymphedema. In 2007 the Axillary Reverse Mapping (ARM) was introduced to limit the invasiveness in the axilla during breast cancer surgery. It is hypothesized that ARM is able to limit the incidence of breast cancer related lymphedema (BCRL) considerably. This systematic review aims to answer the following research questions: (1) which approaches for ARM are described? (2) Is ARM surgical feasible and oncological safe? (3) Does ARM decrease the incidence of lymphedema after sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND)? In total 27 papers were retrieved using four electronic databases (PubMed, Web of Science, Medline and Cochrane clinical trials; assessed until May 13, 2015. The level of evidence of these studies was low (mostly level 3). Therefore the conclusions are that the ARM procedure is feasible although ARM-node rates have a broad range. Additionally, from a theoretical point there is a clear benefit from ARM in terms of lymphedema prevention. From a practical point there is little scientific data to support this due to the lack of studies; and especially because of the different methods and definitions for lymphedema used in the different studies.


Building a taxonomy of integrated palliative care initiatives: results from a focus group.
Empirical evidence suggests that integrated palliative care (IPC) increases the quality of care for palliative patients and supports professional caregivers. Existing IPC initiatives in Europe vary in their design and are hardly comparable. InSuP-C, a European Union research project, aimed to build a taxonomy of IPC initiatives applicable across diseases, healthcare sectors and systems. The taxonomy of IPC initiatives was developed in cooperation with an international and multidisciplinary focus group of 18 experts. Subsequently, a consensus meeting of 10 experts revised a preliminary taxonomy and adopted the final classification system. Consisting of eight categories, with two to four items each, the taxonomy covers the process and structure of IPC initiatives. If two items in at least one category apply to an initiative, a minimum level of integration is assumed to have been reached. Categories range from the type of initiative (items: pathway, model or guideline) to patients' key contact (items: non-pc specialist, pc specialist, general practitioner). Experts recommended the inclusion of two new categories: level of care (items: primary, secondary or tertiary) indicating at which stage palliative care is integrated and primary focus of intervention describing IPC givers' different roles (items: treating function, advising/consulting or training) in the care process. Empirical studies are required to investigate how the taxonomy is used in practice and whether it covers the reality of patients in need of palliative care. The InSuP-C project will test this taxonomy empirically in selected initiatives using IPC.


Absence of biallelic TCRgamma deletion predicts early treatment failure in pediatric T-cell acute lymphoblastic leukemia.
To identify children with T-cell acute lymphoblastic leukemia (T-ALL) at high risk of induction chemotherapy failure by using DNA copy number analysis of leukemic cells collected at diagnosis. Array comparative genomic hybridization (CGH) was performed on genomic DNA extracted from diagnostic lymphoblasts from 47 children with T-ALL treated on Children's Oncology Group Study P9404 or Dana-Farber Cancer Institute Protocol 00-01. These samples represented nine patients who did not achieve an initial complete remission, 13 who relapsed, and 25 who became long-term, event-free survivors. The findings were confirmed in an independent cohort of patients by quantitative DNA polymerase chain reaction (DNA-PCR), an assay that is well suited for clinical application. Analysis of the CGH findings in patients in whom induction chemotherapy failed compared with those in whom induction chemotherapy was successful identified the absence of biallelic TCRgamma locus deletion (ABD), a characteristic of early thymocyte precursors before V(D)J recombination, as the most robust predictor of induction failure (P < .001). This feature was also associated with markedly inferior event-free (P = .002) and overall survival (P < .001) rates: 25% versus 58% and 25% versus 72%, respectively. Using a rapid and inexpensive quantitative DNA-PCR assay, we validated ABD as a predictor of a poor response to induction chemotherapy in an independent series of patients. Lymphoblasts from children with T-ALL should be evaluated at diagnosis for deletion within the TCRgamma locus. Patients lacking biallelic deletion, which confers a high probability of induction failure with contemporary therapy, should be assigned to alternative therapy in the context of a prospective clinical trial.


DL-myo-inositol 1,4,5-trisphosphorothioate mobilizes intracellular calcium in Swiss 3T3 cells and Xenopus oocytes.
The initial water-soluble product of receptor-stimulate polyphosphoinositide hydrolysis, D-myo-inositol 1,4,5-trisphosphate is now accepted as the second messenger that stimulates release of Ca2+ from intracellular pools. We report here the first examples of Ca2+ release by a novel phosphatase-resistant inositol trisphosphate analogue, DL-myo-inositol 1,4,5-trisphosphorothioate, in Swiss 3T3 cells and Xenopus oocytes. L-myo-inositol 1,4,5-trisphosphate was inactive in the latter system.


Chronic voluntary oral methamphetamine induces deficits in spatial learning and hippocampal protein kinase Mzeta with enhanced astrogliosis and cyclooxygenase-2 levels.
Methamphetamine (MA) is an addictive drug with neurotoxic effects on the brain producing cognitive impairment and increasing the risk for neurodegenerative disease. Research has focused largely on examining the neurochemical and behavioral deficits induced by injecting relatively high doses of MA [30 mg/kg of body weight (bw)] identifying the upper limits of MA-induced neurotoxicity. Accordingly, we have developed an appetitive mouse model of voluntary oral MA administration (VOMA) based on the consumption of a palatable sweetened oatmeal mash containing a known amount of MA. This VOMA model is useful for determining the lower limits necessary to produce neurotoxicity in the short-term and long-term as it progresses over time. We show that mice consumed on average 1.743 mg/kg bw/hour during 3 hours, and an average of 5.23 mg/kg bw/day over 28 consecutive days on a VOMA schedule. Since this consumption rate is much lower than the neurotoxic doses typically injected, we assessed the effects of long-term chronic VOMA on both spatial memory performance and on the levels of neurotoxicity in the hippocampus. Following 28 days of VOMA, mice exhibited a significant deficit in short-term spatial working memory and spatial reference learning on the radial 8-arm maze (RAM) compared to controls. This was accompanied by a significant decrease in memory markers protein kinase Mzeta (PKMζ), calcium impermeable AMPA receptor subunit GluA2, and the post-synaptic density 95 (PSD-95) protein in the hippocampus. Compared to controls, the VOMA paradigm also induced decreases in hippocampal levels of dopamine transporter (DAT) and tyrosine hydroxylase (TH), as well as increases in dopamine 1 receptor (D1R), glial fibrillary acidic protein (GFAP) and cyclooxygenase-2 (COX-2), with a decrease in prostaglandins E2 (PGE2) and D2 (PGD2). These results demonstrate that chronic VOMA reaching 146 mg/kg bw/28d induces significant hippocampal neurotoxicity. Future studies will evaluate the progression of this neurotoxic state.


Impact of an insecticide changes with amount of leaf litter input: implications for amphibian populations.
Changes in percentage of forest cover can influence nutrient levels in aquatic systems and change abiotic conditions that may influence species. The authors examined how increasing amounts of leaf litter influenced toxicity of the insecticide carbaryl using larval green frogs (Rana clamitans) in outdoor mesocosm ponds. Insecticides can have direct negative effects on individual physiology and behavior and indirect effects on the food web, which can result in trophic cascades. They predicted that direct effects of the insecticide would dominate when nutrients were low (resulting in negative impacts on amphibian development and survival), whereas indirect effects could offset direct effects when nutrients were more abundant through a trophic cascade that leads to more food for tadpoles (resulting in positive effects on amphibian development and survival). The authors found support for this hypothesis: first, total green frog survival was greatest with increased leaf litter input in the presence of carbaryl. Additionally, most green frogs that reached metamorphosis were from ponds with high leaf litter input and carbaryl. Second, the impact of carbaryl on developmental stage and tadpole mass varied depending on the amount of leaf litter present. With high amounts of leaf litter, carbaryl had a positive impact on development and growth; in contrast, with low amounts of leaf litter, carbaryl had negative or no effects on tadpole development and mass. The present study suggests that differences in nutrient levels between ponds exposed to pesticides could play a role in amphibian population dynamics.


Combination antiretroviral therapy and recent declines in AIDS incidence and mortality.
The reasons for recent declines in AIDS incidence and mortality may include advances in treatment, but these may be confounded by earlier declines in the incidence of human immunodeficiency virus (HIV) infection. To determine whether the declines in AIDS and mortality may, in part, stem from wider use of combination antiretroviral therapy, 622 HIV-positive men with well-characterized dates of seroconversion were followed. In this group, combination therapy came into widespread use in only 1996. In a Cox proportional hazards model, the 1996 calendar period was significantly associated with slower progression to AIDS (relative hazard [RH]=0. 19, 95% confidence interval [CI], 0.05-0.69, P=.01) and death (RH=0. 45, 95% CI, 0.21-0.95, P=.04). Declines in incidence of HIV infection, changes in HIV virulence, and end-point underreporting cannot fully explain the decline in AIDS and death in 1996. The introduction of combination antiretroviral therapy as the standard of care may already have had measurable effects.


Glucagon-like peptide-1 receptor knockout mice are protected from high-fat diet-induced insulin resistance.
Glucagon-like peptide-1 augments nutrient-stimulated insulin secretion. Chow-fed mice lacking the glucagon-like peptide-1 receptor (Glp1r) exhibit enhanced insulin-stimulated muscle glucose uptake but impaired suppression of endogenous glucose appearance (endoRa). This proposes a novel role for the Glp1r to regulate the balance of glucose disposal in muscle and liver by modulating insulin action. Whether this is maintained in an insulin-resistant state is unknown. The present studies tested the hypothesis that disruption of Glp1r expression overcomes high-fat (HF) diet-induced muscle insulin resistance and exacerbates HF diet-induced hepatic insulin resistance. Mice with a functional disruption of the Glp1r (Glp1r-/-) were compared with wild-type littermates (Glp1r+/+) after 12 wk on a regular chow diet or a HF diet. Arterial and venous catheters were implanted for sampling and infusions. Hyperinsulinemic-euglycemic clamps were performed on weight-matched male mice. [3-(3)H]glucose was used to determine glucose turnover, and 2[14C]deoxyglucose was used to measure the glucose metabolic index, an indicator of glucose uptake. Glp1r-/- mice exhibited increased glucose disappearance and muscle glucose metabolic index on either diet. This was associated with enhanced activation of muscle Akt and AMP-activated protein kinase and reduced muscle triglycerides in HF-fed Glp1r-/- mice. Chow-fed Glp1r-/- mice exhibited impaired suppression of endoRa and hepatic insulin signaling. In contrast, HF-fed Glp1r-/- mice exhibited improved suppression of endoRa and hepatic Akt activation. This was associated with decreased hepatic triglycerides and impaired activation of sterol regulatory element-binding protein-1. These results show that mice lacking the Glp1r are protected from HF diet-induced muscle and hepatic insulin resistance independent of effects on total fat mass.


DuPont/HFM Forum on carpet in health care facilities. Roundtable discussion.
DuPont and Health Facilities Management magazine invited 20 national experts to Dalton, GA--the carpet-manufacturing capital of the world--on May 13 to take part in DuPont's first-ever Forum on Carpet in Health Care Facilities. During the two-hour roundtable discussion, moderated by DuPont's C. Jack Murph and HFM's Michael Hemmes, end-users, interior designers and mill representatives talked about the aesthetic, economic and performance aspects of using carpet in health care settings. Here's an edited version of what they said.


An exploratory spatial analysis of pneumonia and influenza hospitalizations in Ontario by age and gender.
Pneumonia and influenza represent a significant public health burden in Canada and abroad. Knowledge of how this burden varies geographically provides clues to understanding the determinants of these illnesses, and insight into the effective management of health-care resources. We conducted a retrospective, population-based, ecological-level study to assess age- and gender-specific spatial patterns of pneumonia and influenza hospitalizations in the province of Ontario, Canada from 1992 to 2001. Results revealed marked variability in hospitalization rates by age, as well as clear and statistically significant patterns of high rates in northern rural counties and low rates in southern urban counties. A moderate yet significant level of positive spatial autocorrelation (Moran's I=0.21, P<0.05) was found in the global data, with significant, age-specific clusters of high values or 'hot spots' identified in several northern counties. Findings illustrate the need for geographically focused prevention strategies, and resource and service allocation policies informed by regional and population-specific demands.


Defect Dynamics in 2-D MoS2 Probed by Using Machine Learning, Atomistic Simulations, and High-Resolution Microscopy.
Structural defects govern various physical, chemical, and optoelectronic properties of two-dimensional transition-metal dichalcogenides (TMDs). A fundamental understanding of the spatial distribution and dynamics of defects in these low-dimensional systems is critical for advances in nanotechnology. However, such understanding has remained elusive primarily due to the inaccessibility of (a) necessary time scales via standard atomistic simulations and (b) required spatiotemporal resolution in experiments. Here, we take advantage of supervised machine learning, in situ high-resolution transmission electron microscopy (HRTEM) and molecular dynamics (MD) simulations to overcome these limitations. We combine genetic algorithms (GA) with MD to investigate the extended structure of point defects, their dynamical evolution, and their role in inducing the phase transition between the semiconducting (2H) and metallic (1T) phase in monolayer MoS2. GA-based structural optimization is used to identify the long-range structure of randomly distributed point defects (sulfur vacancies) for various defect densities. Regardless of the density, we find that organization of sulfur vacancies into extended lines is the most energetically favorable. HRTEM validates these findings and suggests a phase transformation from the 2H-to-1T phase that is localized near these extended defects when exposed to high electron beam doses. MD simulations elucidate the molecular mechanism driving the onset of the 2H to 1T transformation and indicate that finite amounts of 1T phase can be retained by increasing the defect concentration and temperature. This work significantly advances the current understanding of defect structure/evolution and structural transitions in 2D TMDs, which is crucial for designing nanoscale devices with desired functionality.


Octenidine Hydrochloride in Hydatid Disease.
Hydatid disease is still endemic in many devoloping countries and continues to be an important cause of morbidity. The objective of this study was to determine the in vitro scolicidal effects of octenidine hydrochloride in different concentrations using different exposure times. After hydatid cyst liquid was left to precipitate for 1 h to obtain cystic sand, various concentrations of octenidine (undiluted, 1% and 0.1% diluted) were added to concentrated hydatid cyst sediments for 5, 10, 15, 20, 25, 30, 45, and 60 min, and scolicidal effects of octenidine were compared with 20% saline and control group for the same times. It was found that undiluted octenidine had a strong scolicidal effect at 15 min compared to saline at 20%. One percent octenidine had a scolicidal effect at 30 min. However, 0.1% octenidine did not have enough scolicidal effect in 1 h. It was concluded that undiluted and 1% diluted octenidine might be used for scolicidal purpose in the treatment of hydatid disease.


Neutrophil attractant protein-1-immunoglobulin G immune complexes and free anti-NAP-1 antibody in normal human serum.
After obtaining data indicating the presence of a neutrophil attractant protein-1 (NAP-1)-IgG complex in normal human serum, we developed sandwich ELISAs that could quantify NAP-1 and NAP-1-IgG in mixtures of the two moieties. The ELISA for free NAP-1 used a monoclonal capture antibody that did not bind NAP-1-IgG. The ELISA for NAP-1-IgG was based on omission of the anti-NAP-1 detection antibody (required for the free NAP-1 ELISA) and on interaction of phosphatase-conjugated anti-human IgG with the human NAP-1-IgG complex. Gel filtration of immunoaffinity-purified NAP-1-IgG showed that the bulk of the complex comprised a single IgG. Binding between NAP-1 and antibody is strong, since 8 M urea at neutral or alkaline pH did not release NAP-1. However, at pH 2.0 in 9 M urea approximately 15% of the total NAP-1 could be dissociated from the complex. NAP-1-IgG was detected in 18 of 26 sera from normal humans. The mean serum concentration was 58 ng of IgG-bound NAP-1/ml, with an SEM of 16 and a range from undetectable to 247 ng/ml. NAP-1-IgG concentrations in paired sera drawn at a 1-mo interval were remarkably constant. Using an ELISA for free NAP-1 with a detection limit of 200 pg/ml, we found no free NAP-1 in the 26 sera. Free anti-NAP-1-IgG autoantibody was found in 9 of 26 sera by direct ELISA. IgG anti-NAP-1 of all nine sera was polyclonal, comprising both kappa and lambda isotypes; predominant subclasses were IgG2 and IgG3. NAP-1-IgG did not compete with 125I-NAP-1 for binding to neutrophils, which suggests that IgG anti-NAP-1 is a molecular trap that prevents binding of NAP-1 to neutrophils after it diffuses from production sites into the circulation.


Lack of GNAQ and GNA11 Germ-Line Mutations in Familial Melanoma Pedigrees with Uveal Melanoma or Blue Nevi.
Approximately 10% of melanoma cases are familial, but only 25-40% of familial melanoma cases can be attributed to germ-line mutations in the CDKN2A - the most significant high-risk melanoma susceptibility locus identified to date. The pathogenic mutation(s) in most of the remaining familial melanoma pedigrees have not yet been identified. The most common mutations in nevi and sporadic melanoma are found in BRAF and NRAS, both of which result in constitutive activation of the MAPK pathway. However, these mutations are not found in uveal melanomas or the intradermal melanocytic proliferations known as blue nevi. Rather, multiple studies report a strong association between these lesions and somatic mutations in Guanine nucleotide-binding protein G(q) subunit alpha (GNAQ), Guanine nucleotide-binding protein G(q) subunit alpha-11 (GNA11), and BRCA1-associated protein-1 (BAP1). Recently, germ-line mutations in BAP1, the gene encoding a tumor suppressing deubiquitinating enzyme, have been associated with predisposition to a variety of cancers including uveal melanoma, but no studies have examined the association of germ-line mutations in GNAQ and GNA11 with uveal melanoma and blue nevi. We have now done so by sequencing exon 5 of both of these genes in 13 unique familial melanoma pedigrees, members of which have had either uveal or cutaneous melanoma and/or blue nevi. Germ-line DNA from a total of 22 individuals was used for sequencing; however no deleterious mutations were detected. Nevertheless, such candidate gene studies and the discovery of novel germ-line mutations associated with an increased MM susceptibility can lead to a better understanding of the pathways involved in melanocyte transformation, formulation of risk assessment, and the development of specific drug therapies.


Population genetics and phylogenetic analysis of Colorado Borrelia burgdorferi.
Borrelia burgdorferi is transmitted in an enzootic cycle in Colorado between the tick Ixodes spinipalpis and the woodrat Neotoma mexicana. The genetic relationship of Colorado isolates to other B. burgdorferi isolates is unknown nor have relationships among various Colorado isolates been determined. Portions of the flagellin (fla), 66-kD protein, and outer surface protein A (ospA) genes were amplified from 71 Colorado isolates, screened for genetic variability using single strand conformation polymorphism analysis, and unique alleles were sequenced. Colorado isolates were most similar to tick isolates from California and New York isolate 25015. Genetic distances among Colorado ospA sequences were the same or higher than distances among other isolates whereas distances among fla sequences tended to be the same or lower. The index of association (I(A)) was calculated among all loci as a measure of clonality. The I(A) among Colorado isolates was similar to I(A) previously estimated among other United States isolates.


Radiotelemetry and wildlife: Highlighting a gap in the knowledge on radiofrequency radiation effects.
Radio transmitters and associated devices may induce negative effects that can bias the results of ongoing research. The main documented effects of radio transmitters on animals include reduced survival, decreased productivity, changes in behaviour and movement patterns and a biased sex ratio. The only factors that have claimed responsibility for these possible damages are the weight of the radio transmitter and associated devices, and the attachment type. The electromagnetic radiation produced by radio transmitters has not been considered so far in research. There have been no studies evaluating the effects of non-ionising electromagnetic radiation (radiofrequency signals) necessary for tracking, although the problems found were significantly associated with the length of time that animals had been carrying their radio transmitters. Similar problems as those in radiotracked animals have been found in numerous studies with animals exposed to radiofrequency radiation for a sufficient amount of time. Laboratory scientists investigating the orientation of animals know they have to shield the place where experiments are performed to prevent interference from man-made radiation, as anthropogenic signals may distort the results. It is paradoxical that, at the same time, field scientists investigating the movements and other aspects of animal biology are providing animals with radio transmitters that emit the same type of radiation, since this may affect the results concerning their orientation and movement. This paper identifies gaps in the knowledge that should be investigated in-depth. The possibility that the radiofrequency radiation from radiotracking devices is responsible for the findings should be considered. Considering this factor may allow researchers to best understand the long-term effects found.


Outpatient management of intra-corporeal left ventricular assist device system in children: a multi-center experience.
Little is known about the outcomes of children supported on intracorporeal left ventricular assist device (HVAD), and the feasibility of outpatient management. All centers with pediatric patients discharged from the hospital on the device were identified using company database. A total of 14 centers were contacted, with 9 centers, contributing data retrospectively. From 2011 to 2013, 12 pediatric patients (7 females), mean aged 11.9 ± 2.3 years (range 8-15), mean weight 43 ± 19 kg (range 18-81), mean body surface area 1.3 ± 0.3 m(2) (range 0.76-1.96) were identified. Diagnosis included: dilated cardiomyopathy (CMP) (n = 5), noncompaction CMP (n = 4), toxic CMP (n = 2) and viral CMP (n = 1). Indications for support were permanent support (n = 1), bridge to recovery (n = 1) and bridge to transplantation (n = 10). Prior to HVAD implantation, all patients received intravenous inotropes and two patients were on temporary mechanical support. Overall mortality was 0%. Mean duration of inpatient and outpatient support were 56 (range: 19-95 days) and 290 days (range: 42-790), respectively. Mean readmission rate was 0.02 per patient month (2.1 per patient). No adverse events involving emergency department occurred. Eight children resumed local schooling. Home discharge of children supported on HVAD is feasible and safe. School integration can be achieved. There is wide center variability to discharge practice for children.


Polymorphisms in the circadian expressed genes PER3 and ARNTL2 are associated with diurnal preference and GNβ3 with sleep measures.
Sleep and circadian rhythms are intrinsically linked, with several sleep traits, including sleep timing and duration, influenced by both sleep homeostasis and the circadian phase. Genetic variation in several circadian genes has been associated with diurnal preference (preference in timing of sleep), although there has been limited research on whether they are associated with other sleep measurements. We investigated whether these genetic variations were associated with diurnal preference (Morningness-Eveningness Questionnaire) and various sleep measures, including: the global Pittsburgh Sleep Quality index score; sleep duration; and sleep latency and sleep quality. We genotyped 10 polymorphisms in genes with circadian expression in participants from the G1219 sample (n = 966), a British longitudinal population sample of young adults. We conducted linear regressions using dominant, additive and recessive models of inheritance to test for associations between these polymorphisms and the sleep measures. We found a significant association between diurnal preference and a polymorphism in period homologue 3 (PER3) (P < 0.005, recessive model) and a novel nominally significant association between diurnal preference and a polymorphism in aryl hydrocarbon receptor nuclear translocator-like 2 (ARNTL2) (P < 0.05, additive model). We found that a polymorphism in guanine nucleotide binding protein beta 3 (GNβ3) was associated significantly with global sleep quality (P < 0.005, recessive model), and that a rare polymorphism in period homologue 2 (PER2) was associated significantly with both sleep duration and quality (P < 0.0005, recessive model). These findings suggest that genes with circadian expression may play a role in regulating both the circadian clock and sleep homeostasis, and highlight the importance of further studies aimed at dissecting the specific roles that circadian genes play in these two interrelated but unique behaviours.


Non-invasive prognostic protein biomarker signatures associated with colorectal cancer.
The current management of colorectal cancer (CRC) would greatly benefit from non-invasive prognostic biomarkers indicative of clinicopathological tumor characteristics. Here, we employed targeted proteomic profiling of 80 glycoprotein biomarker candidates across plasma samples of a well-annotated patient cohort with comprehensive CRC characteristics. Clinical data included 8-year overall survival, tumor staging, histological grading, regional localization, and molecular tumor characteristics. The acquired quantitative proteomic dataset was subjected to the development of biomarker signatures predicting prognostic clinical endpoints. Protein candidates were selected into the signatures based on significance testing and a stepwise protein selection, each within 10-fold cross-validation. A six-protein biomarker signature of patient outcome could predict survival beyond clinical stage and was able to stratify patients into groups of better and worse prognosis. We further evaluated the performance of the signature on the mRNA level and assessed its prognostic value in the context of previously published transcriptional signatures. Additional signatures predicting regional tumor localization and disease dissemination were also identified. The integration of rich clinical data, quantitative proteomic technologies, and tailored computational modeling facilitated the characterization of these signatures in patient circulation. These findings highlight the value of a simultaneous assessment of important prognostic disease characteristics within a single measurement.


Rat liver sinusoidal endothelial cells (LSECs) express functional low density lipoprotein receptor-related protein-1 (LRP-1).
The low density lipoprotein receptor-related protein-1 (LRP-1) is a large, multifunctional endocytic receptor from the LDL receptor family, highly expressed in liver parenchymal cells (PCs), neurons, activated astrocytes, and fibroblasts. The aim of the study was to investigate if liver sinusoidal endothelial cells (LSECs), highly specialized scavenger cells, express LRP-1. To address this question, experiments were performed in vivo and in vitro to determine if receptor associated protein (RAP) and trypsin-activated α(2)-macroglobulin (α(2)M∗) were endocytosed in LSECs. Both ligands were cleared from the circulation mainly by the liver. Hepatocellular distribution of intravenously administered ligands, assessed after magnetic bead cell separation using LSEC- and KC-specific antibodies, showed that PCs contained 93% and 82% of liver-associated (125)I-RAP and (125)I-α(2)M∗, whereas 5% and 11% were associated with LSECs. Uptake of RAP and α(2)M∗ in the different liver cell population in vitro was specific and followed by degradation. The uptake of (125)I-RAP was not inhibited by ligands to known endocytosis receptors in LSECs, while uptake of (125)I-α(2)M∗ was significantly inhibited by RAP, suggesting the involvement of LRP-1. Immunofluorescence using LRP-1 antibody showed positive staining in LSECs. Ligand blot analyses using total cell proteins and (125)I-RAP followed by mass spectrometry further confirmed and identified LRP-1 in LSECs. LSECs express functional LRP-1. An important implication of our findings is that LSECs contribute to the rapid removal of blood borne ligands for LRP-1 and may thus play a role in lipid homeostasis.


Mutual regulation of vasopressin- and oxytocin-induced glucagon secretion in V1b vasopressin receptor knockout mice.
[Arg8]-vasopressin (AVP) and oxytocin (OT) are neurohypophysial hormones which exert various actions, including the control of blood glucose, in some peripheral tissues. To investigate the type of receptors involved in AVP- and OT-induced glucagon secretion, we investigated the effect of these peptides on glucagon secretion in islets of wild-type (V1bR+/+) and vasopressin V1b receptor knockout (V1bR-/-) mice. AVP-induced glucagon secretion was significantly inhibited by the selective V1b receptor antagonist, SSR149415 (30%), and OT-induced glucagon secretion by the specific OT receptor antagonist, d(CH2)5[Tyr(Me)2, Thr4, Tyr-NH(2)(9)]OVT (CL-14-26) (45%), in islets of V1bR+/+mice. AVP- and OT-induced glucagon secretions were not by the antagonist of each, but co-incubation with both 10(-6) M SSR149415 and 10(-6) M CL-14-26 further inhibited AVP- and OT-induced glucagon secretions in islets of V1bR+/+ mice (57 and 69% of the stimulation values respectively). In addition, both AVP and OT stimulated glucagon secretion with the same efficacy in V1bR-/- mice as in V1bR+/+ mice. AVP- and OT-induced glucagon secretion in V1bR-/- mice was significantly inhibited by CL-14-26. These results demonstrate that V1b receptors can mediate OT-induced glucagon secretion and OT receptors can mediate AVP-induced glucagon secretion in islets from V1bR+/+mice in the presence of a heterologous antagonist, while AVP and OT can stimulate glucagon secretion through the OT receptors in V1bR-/-mice, suggesting that the other receptor can compensate when one receptor is absent.


Ocular perfusion pressure and the incidence of glaucoma: real effect or artifact? The Rotterdam Study.
To determine the association between the ocular perfusion pressure (OPP; essentially the difference between the blood pressure and the intraocular pressure [IOP]) and incident open-angle glaucoma (OAG). A subset of 3882 participants of the population-based Rotterdam Study for whom data from ophthalmic examinations at baseline and follow-up and blood pressure measurements at baseline were available, and who did not have OAG at baseline, were included. Associations between the mean, systolic and diastolic OPP, and incident OAG were assessed using Cox regression models adjusted for age and sex, with and without adjustment for IOP. During a mean follow-up of 9.8 years, 103 participants (2.7%) developed OAG. The association between the mean OPP and incident OAG was not significant (hazard ratio 0.995 per mm Hg increase in mean OPP; 95% confidence interval 0.971-1.019) when adjusted for IOP, but became significant if not adjusted for IOP (0.968; 0.945-0.992). The systolic and diastolic OPP showed a pattern similar to that of the mean OPP, though less significant. The OPP appears to be associated with incident OAG but this association seems to be due to the fact that the IOP, a strong risk factor for OAG, is part of the OPP, rather than that OPP is an independent OAG risk factor itself.


FTY720 treatment in the convalescence period improves functional recovery and reduces reactive astrogliosis in photothrombotic stroke.
The Sphingosine-1-phosphate (S1P) signaling pathway is known to influence pathophysiological processes within the brain and the synthetic S1P analog FTY720 has been shown to provide neuroprotection in experimental models of acute stroke. However, the effects of a manipulation of S1P signaling at later time points after experimental stroke have not yet been investigated. We examined whether a relatively late initiation of a FTY720 treatment has a positive effect on long-term neurological outcome with a focus on reactive astrogliosis, synapses and neurotrophic factors. We induced photothrombotic stroke (PT) in adult C57BL/6J mice and allowed them to recover for three days. Starting on post-stroke day 3, mice were treated with FTY720 (1 mg/kg b.i.d.) for 5 days. Behavioral outcome was observed until day 31 after photothrombosis and periinfarct cortical tissue was analyzed using tandem mass-spectrometry, TaqMan®analysis and immunofluorescence. FTY720 treatment results in a significantly better functional outcome persisting up to day 31 after PT. This is accompanied by a significant decrease in reactive astrogliosis and larger post-synaptic densities as well as changes in the expression of vascular endothelial growth factor α (VEGF α). Within the periinfarct cortex, S1P is significantly increased compared to healthy brain tissue. Besides its known neuroprotective effects in the acute phase of experimental stroke, the initiation of FTY720 treatment in the convalescence period has a positive impact on long-term functional outcome, probably mediated through reduced astrogliosis, a modulation in synaptic morphology and an increased expression of neurotrophic factors.


Associations of symptoms and quality of life with outcomes in patients with atrial fibrillation.
We aimed to investigate changes in atrial fibrillation (AF)-related symptoms and quality of life (QoL) over time, and their impact on prognosis. We prospectively followed 3836 patients with known AF for a mean of 3.7 years. Information on AF-related symptoms and QoL was obtained yearly. The primary end point was a composite of stroke or systemic embolism. Main secondary end points included stroke subtypes, all-cause mortality, cardiovascular death, hospitalisation for congestive heart failure (CHF), myocardial infarction and major bleeding. We assessed associations using multivariable, time-updated Cox proportional hazards models. Mean age was 72 years, 72% were male. Patients with AF-related symptoms (66%) were younger (70 vs 74 years, p<0.0001), more often had paroxysmal AF (56% vs 37%, p<0.0001) and had lower QoL (71 vs 72 points, p=0.009). The incidence of the primary end point was 1.05 and 1.02 per 100 person-years in patients with and without symptoms, respectively. The multivariable adjusted HR (aHR) (95% CIs) for the primary end point was 1.11 (0.77 to 1.59; p=0.56) for AF-related symptoms. AF-related symptoms were not associated with any of the secondary end points. QoL was not significantly related to the primary end point (aHR per 5-point increase 0.98 (0.94 to 1.03; p=0.37)), but was significantly related to CHF hospitalisations (0.92 (0.90 to 0.94; p<0.0001)), cardiovascular death (0.90 (0.86 to 0.95; p<0.0001)) and all-cause mortality (0.88 (0.86 to 0.90; p<0.0001)). AF-related symptoms were not associated with adverse outcomes and should therefore not be the basis for prognostic treatment decisions. QoL was strongly associated with CHF, cardiovascular death and all-cause mortality.


Fusion between large phagocytic vesicles: targeting of yeast and other particulates to phagolysosomes that shelter the bacterium Coxiella burnetii or the protozoan Leishmania amazonensis in Chinese hamster ovary cells.
This report examines the fusion of phagocytic vesicles with the large phagolysosome-like vacuoles induced in Chinese hamster ovary cells by the bacterium Coxiella burnetti or the Protozoan flagellate Leishmania amazonensis. Infection by these organisms is compatible with cell survival and multiplication. Fusion was inferred from the transfer of microscopically identifiable particles from donor to target vesicles. Donor vesicles contained heat-killed yeast, zymosan, beta-glucan or latex beads taken up by the host cells. Yeast and zymosan were also coated with Concanavalin A to increase their uptake by the cells (Goldman, R., Exp. Cell Res. 104, 325-334, 1977). Particle localization, routinely ascertained by phase-contrast microscopy, was confirmed by confocal laser fluorescence and by transmission electron microscopy. Coxiella vacuoles admitted all the particles tested and transfer took place whether the particles were given to the cells prior to or after infection. Transfer of uncoated or Concanavalin-A-coated yeast or zymosan was dependent on the number of particles ingested and on the incubation period (between 2 and 24 hours). Furthermore, the transfer step was quite efficient, since over 85% of the particles ingested entered Coxiella vacuoles at all particle to cell ratios examined. The fraction of uncoated or Concanavalin-A-coated yeast or zymosan transferred to Leishmania vacuoles was consistently lower and diminished at higher particle loads. In addition, only rarely did latex beads enter these vacuoles. The models proposed may be useful for the delineation of biochemical and molecular mechanisms involved in the fusion of large phagocytic vesicles and the modulation of the latter by cellular and pathogen-derived signals.


Sexual-orientation disparities in substance use in emerging adults: a function of stress and attachment paradigms.
More lesbian, gay, and bisexual (LGB) youths than heterosexuals report substance use. We examined a theoretical model to understand these disparities in lifetime and past-year substance use by means of stress and attachment paradigms, using the longitudinal Growing Up Today Study (GUTS) and Nurses' Health Study II (NHSII). GUTS participants are the children of participants in NHSII; thus, child and maternal data are available. In addition, GUTS contains siblings, allowing for comparisons of LGB and heterosexual siblings. Of 5,647 GUTS youths (M = 20.6 years old in 2005), 1.6% were lesbian/gay (LG), 1.6% bisexual (BI), 9.9% mostly heterosexual (MH), and 86.9% completely heterosexual (CH). After adjusting for sibling clustering in GUTS and covariates, significantly more sexual minorities (LGs, BIs, and MHs) than CHs reported lifetime and past-year smoking, nonmarijuana illicit drug use, and prescription drug misuse. More sexual minorities also reported marijuana use in the past year. The relations between sexual orientation and substance use were moderated by the stress markers: As mother's discomfort with homosexuality increased, more BIs and MHs than CHs used substances. As childhood gender nonconforming behaviors increased, more LGs than CHs used substances. The relations between sexual orientation and substance use were mediated by attachment and maternal affection (percent of effect mediated ranged from 5.6% to 16.8%% for lifetime substance use and 4.9% to 24.5% for past-year use). In addition, sibling comparisons found that sexual minorities reported more substance use, more childhood gender nonconforming behaviors, and less secure attachment than CH siblings; mothers reported less affection for their sexual minority than CH offspring. The findings indicate the importance of stress and attachment paradigms for understanding sexual-orientation disparities in substance use.


In-depth analysis of pH-dependent mechanisms of electromechanical reshaping of rabbit nasal septal cartilage.
Electromechanical reshaping (EMR) involves reshaping cartilage by mechanical deformation and delivering electric current to the area around the bend axis, causing local stress relaxation and permanent shape change. The mechanism of EMR is currently unclear, although preliminary studies suggest that voltage and application time are directly related to the concentration and diffusion of acid-base products within the treated tissue with little heat generation. This study aims to characterize local tissue pH changes following EMR and to demonstrate that local tissue pH changes are correlated with tissue damage and shape change. Ex vivo animal study involving EMR of rabbit nasal septal cartilage and biochemical estimation of tissue pH changes. The magnitude and diffusion of acid-base chemical products in control (0V, 2 minutes), shape change (4V, 4 minutes; 6V, 1, 2, 4 minutes; 8V, 1, 2 minutes), and tissue damage (8V, 4, 5 minutes; 10V, 4, 5 minutes) parameters following EMR are approximated by analyzing local pH changes after pH indicator application. There is a direct relationship between total charge transfer and extent of acid-base product diffusion (P <0.05). A "pH transition zone" is seen surrounding the bend apex above 8V, 2 minutes. Colorimetric analysis suggests that small local pH changes (10(-8) hydrogen ions) are at least partly implicated in clinically efficacious EMR. These results provide additional insight into the translational applications of EMR, particularly the relationship among pH changes, shape change, and tissue injury, and are integral in optimizing this promising technology for clinical use.


Social class, risk factors, and stroke incidence in men and women: a prospective study in the European prospective investigation into cancer in Norfolk cohort.
The purpose of this study was to investigate the association between occupational social class and stroke incidence and the extent to which classical, lifestyle, and psychosocial risk factors may explain such relationships. A prospective population study was conducted of 22488 men and women aged 39 to 79 years living in the general community in Norfolk, UK, recruited in 1993 to 1997 and followed up for stroke incidence to 2007. An inverse relationship was observed between social class and stroke incidence with an age- and sex-adjusted hazard ratio for social Class V compared with I of 2.62 (95% CI, 1.63 to 4.22; P=0.001). Adjusting for classical (systolic blood pressure, total blood cholesterol, smoking, history of diabetes, and body mass index), lifestyle (plasma vitamin C levels, alcohol intake, and physical activity), and psychosocial (5-item version of the Mental Health Inventory) risk factors had little effect, and a socioeconomic differential was still apparent: hazard ratio for social Class V compared with I of 2.55 (95% CI, 1.34 to 4.85, P=0.004 for comparison of V to I). Stroke incidence increased with lower social class in both men and women. Adjustment for a comprehensive range of classical, lifestyle, and psychosocial risk factors did not explain the socioeconomic differential in stroke incidence. If we are to reduce inequalities in health, further understanding of the mechanisms underlying the association is needed.


Mitochondrial responses of normal and injured human skin fibroblasts following low level laser irradiation--an in vitro study.
Laser irradiation has proved to be very efficient in speeding and improving the quality of healing in pathological conditions of diverse etiologies. However, the mechanisms by which the beneficial effects are attained are not clear. Mitochondria are the primary phototargets during irradiation. The study aimed to establish if laser irradiation had an effect on hypoxic and acidotic cells. The study also aimed to use existing information regarding the possible mechanism of action (established in wounded cells) and apply these principles to acidic and hypoxic irradiated cells to determine whether laser has a stimulatory or inhibitory effect. Cell cultures were modified to simulate conditions of hypoxia (hypoxic gas mixture 95% N2 and 5% O2) and acidosis (pH 6.7) whereas the central scratch model was used to simulate a wound. Cells were irradiated with a helium-neon (632.8 nm, 3 mW cm(-2)) laser using 5 or 16 J cm(-2) on days 1 and 4. Mitochondrial responses were measured 1 or 24 h after laser irradiation by assessing changes in mitochondrial membrane potential (MMP), cyclic AMP, intracellular Ca2+ and adenosine triphosphate (ATP) cell viability. Hypoxia and acidosis significantly reduced MMP when compared with normal nonirradiated control cells. Wounded, hypoxic and acidotic cells irradiated with 5 J cm(-2) showed an increase in mitochondrial responses when compared with nonirradiated cells while 16 J cm(-2) showed a significant decrease. The study confirmed that laser irradiation with 5 J cm(-2) stimulated an increase in intracellular Ca2+ which resulted in an increase in MMP, ATP and cAMP, which ultimately results in photobiomodulation to restore homeostasis of injured cells.


Characteristics of the built environment in relation to objectively measured physical activity among Mexican adults, 2011.
The built environment correlates of physical activity are documented in high-income countries but have yet to be studied among Mexican adults. Our objectives were to assess the associations between characteristics of the built environment and physical activity among adults in Cuernavaca, Mexico, and to examine potential moderation by perceived park and neighborhood safety. We conducted a population-based study of adults in Cuernavaca, Mexico, in 2011 (N = 677). Participants wore Actigraph GT3X accelerometers for 7 days. We used geographic information systems (GIS) to generate 500-m- and 1-km-buffer-based measures of net residential density, proportion of commercial land use, land-use mix, connectivity, walkability, and number of parks and transit routes. We also obtained data on distance to the nearest park with GIS. Perceived neighborhood and park safety were self-reported. We created quartile-based categories for all built environment characteristics and ran linear regression models to estimate the association between each characteristic and total weekly moderate-to-vigorous physical activity (MVPA) and MVPA within 10-minute bouts. Walkability was inversely related to total weekly minutes of MVPA (1-km buffer, -46.9 [standard error, 20.0]; P = .03) and weekly minutes of MVPA within bouts (500-m buffer, -31.5 [12.9]; P = .02). The number of transit routes in the 500-m buffer was inversely related to total weekly minutes of MVPA (-23.8 [10.6]; P = .04). Perception of park safety moderated the association between physical activity and having a park intersect the 500-m buffer. Our findings contrast with those from high-income countries, suggesting that environmental programs and policies to increase physical activity in Mexican cities cannot be adapted from high-income countries without considering the local context.


Total Laparoscopic Reversal ALPPS.
Associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) allows R0 resection even for patients with extremely small future liver remnants. The ALPPS procedure was initially described for two-stage right trisectionectomy. Reversal ALPPS is a denomination in which the future liver remnant is the right posterior section of the liver. A 42-year-old woman with colorectal metastases in all segments except segment 1 underwent chemotherapy with objective response and was referred for surgical treatment. The computed tomography (CT) scan showed a predominance of metastases in the left liver and in the right anterior section. The right posterior section had three metastases. The plan was to perform a laparoscopic reversal ALPPS (left portal vein ligation combined with in situ splitting in a two-stage left trisectionectomy). Three metastases in the right posterior section were resected, followed by liver partition and left portal vein ligature. The CT scan showed a 70 % increase in the future liver remnant. The second stage constituted left trisectionectomy. At laparoscopy after division of adhesions, the left Glissonian pedicle was divided with an endostapler. A stapler also was used to transect the left and middle hepatic veins, and the specimen was removed through a suprapubic incision. The operative times were respectively 5 and 3 h, and the patient was discharged on days 4 and 5, respectively. No blood transfusion or intensive care unit stay was necessary. At this writing, the patient shows no evidence of the disease 18 months after the procedure. Reversal laparoscopic ALPPS is feasible and safe. Laparoscopy is useful for decreasing blood loss and optimizing visualization during liver transection.


EQ-5D in Central and Eastern Europe: 2000-2015.
Cost per quality-adjusted life year data are required for reimbursement decisions in many Central and Eastern European (CEE) countries. EQ-5D is by far the most commonly used instrument to generate utility values in CEE. This study aims to systematically review the literature on EQ-5D from eight CEE countries. An electronic database search was performed up to 1 July 2015 to identify original EQ-5D studies from the countries of interest. We analysed the use of EQ-5D with respect to clinical areas, methodological rigor, population norms and value sets. We identified 143 studies providing 152 country-specific results with a total sample size of 81,619: Austria (n = 11), Bulgaria (n = 6), Czech Republic (n = 18), Hungary (n = 47), Poland (n = 51), Romania (n = 2), Slovakia (n = 3) and Slovenia (n = 14). Cardiovascular (21 %), neurologic (17 %), musculoskeletal (15 %) and endocrine, nutritional and metabolic diseases (13 %) were the most frequently studied clinical areas. Overall, 112 (78 %) of the studies reported EQ VAS results and 86 (60 %) EQ-5D index scores, of which 27 (31 %) did not specify the applied tariff. Hungary, Poland and Slovenia have population norms. Poland and Slovenia also have a national value set. Increasing use of EQ-5D is observed throughout CEE. The spread of health technology assessment activities in countries seems to be reflected in the number of EQ-5D studies. However, improvement in informed use and methodological quality of reporting is needed. In jurisdictions where no national value set is available, in order to ensure comparability we recommend to apply the most frequently used UK tariff. Regional collaboration between CEE countries should be strengthened.


Vascular endothelial growth factor antagonism restores epithelial barrier dysfunction via affecting zonula occludens proteins.
Epithelial barrier dysfunction is important in the pathogenesis of asthma and allergic responses, and is therefore a therapeutic target. The aim of the present study was to investigate the effects of dexamethasone, a classic therapeutic agent, an anti-tumor necrosis factor agent (etanercept), which is used to treat difficult cases of asthma, and an anti-vascular endothelial growth factor (VEGF) agent (bevacizumab), which is an angiogenesis inhibitor, on zonula occludens (ZO) proteins in an experimental asthma model. The experimental model of asthma was developed using intraperitoneal (IP) and inhaled administration of ovalbumin in 38 BALB/c mice, which were divided into four groups. The control group (n=6) did not receive any treatment, while the four remaining groups (n=8 per group) received an IP injection of saline, etanercept, bevacizumab or dexamethasone, respectively. Occludin, claudin and junctional adhesion molecule (JAM) were immunohistochemically stained in the left middle lobe samples using an indirect avidin-peroxidase method, after which the staining was semiquantified with H-scores. Statistically significant differences were observed in the occludin, claudin and JAM H-scores among the four groups (P<0.001). In the untreated asthma, etanercept, bevacizumab and dexamethasone groups, the median H-scores for occludin were 93, 177, 280 and 198, respectively, while the H-scores for claudin were 82, 193.5, 274 and 202.5, respectively, and the median H-scores for JAM were 130, 210, 288 and 210, respectively. Pairwise comparisons revealed that all three ZO protein H-scores were significantly lower in the saline group when compared with each treatment group. However, the H-scores of the ZO proteins were not significantly different between the etanercept and dexamethasone groups. Furthermore, the bevacizumab group exhibited higher H-scores for all the proteins compared with the dexamethasone group. Therefore, antagonism of VEGF with bevacizumab restores the epithelial barrier to a greater extent when compared with dexamethasone treatment. This result may be promising for the development of novel therapeutic agents.


Effects of vagotomy on drinking in the rat.
1. Left or right cervical vagotomy caused elevated intakes of water after hypertonic saline, isoprenaline and polyethylene glycol compared with normal rats. In cervically vagotomized rats sodium nitrite caused a dose-dependent water intake whereas in normal rats it produced a smaller, more variable response. The same procedure also reduced the extent to which drinking following overnight water deprivation was inhibited by plasma volume expansion. Drinking after diazoxide or in response to ligation of the abdominal inferior vena cava was not affected by cervical vagotomy. 2. The antidiuresis that followed isoprenaline treatment was greater in cervically vagotomized than normal rats; that which occurred after caval ligation was unaffected. 3. Subdiaphragmatic vagotomy did not alter water intake in response to hypertonic saline, isoprenaline or polyethylene glycol. 4. It is concluded that nerves within the supradiaphragmatic distribution of the vagi modify water intake in response to changes in the degree of vascular filling.


Pediatric fecal microbiota harbor diverse and novel antibiotic resistance genes.
Emerging antibiotic resistance threatens human health. Gut microbes are an epidemiologically important reservoir of resistance genes (resistome), yet prior studies indicate that the true diversity of gut-associated resistomes has been underestimated. To deeply characterize the pediatric gut-associated resistome, we created metagenomic recombinant libraries in an Escherichia coli host using fecal DNA from 22 healthy infants and children (most without recent antibiotic exposure), and performed functional selections for resistance to 18 antibiotics from eight drug classes. Resistance-conferring DNA fragments were sequenced (Illumina HiSeq 2000), and reads assembled and annotated with the PARFuMS computational pipeline. Resistance to 14 of the 18 antibiotics was found in stools of infants and children. Recovered genes included chloramphenicol acetyltransferases, drug-resistant dihydrofolate reductases, rRNA methyltransferases, transcriptional regulators, multidrug efflux pumps, and every major class of beta-lactamase, aminoglycoside-modifying enzyme, and tetracycline resistance protein. Many resistance-conferring sequences were mobilizable; some had low identity to any known organism, emphasizing cryptic organisms as potentially important resistance reservoirs. We functionally confirmed three novel resistance genes, including a 16S rRNA methylase conferring aminoglycoside resistance, and two tetracycline-resistance proteins nearly identical to a bifidobacterial MFS transporter (B. longum s. longum JDM301). We provide the first report to our knowledge of resistance to folate-synthesis inhibitors conferred by a predicted Nudix hydrolase (part of the folate synthesis pathway). This functional metagenomic survey of gut-associated resistomes, the largest of its kind to date, demonstrates that fecal resistomes of healthy children are far more diverse than previously suspected, that clinically relevant resistance genes are present even without recent selective antibiotic pressure in the human host, and that cryptic gut microbes are an important resistance reservoir. The observed transferability of gut-associated resistance genes to a gram-negative (E. coli) host also suggests that the potential for gut-associated resistomes to threaten human health by mediating antibiotic resistance in pathogens warrants further investigation.


Skin impulse excitation of spinal sensory neurons in developing Xenopus laevis (Daudin) tadpoles.
Responses to gentle touch in young Xenopus tadpoles are mediated by spinal cord sensory Rohon-Beard neurons. Tadpoles also respond to noxious stimuli that elicit 'skin impulses', which propagate between epithelial cells over the whole body surface, somehow entering the CNS to generate a response. After hatching (~48 h post-fertilization), skin impulse signals enter the CNS only via cranial nerves, but previous evidence suggested the possibility of direct entry to the spinal cord before this (~24 h). We have used behavioural and electrophysiological methods to explore the developmental pattern of skin impulse entry into the spinal cord and the involvement of Rohon-Beard neurons. Lesioning confirmed that skin impulse signals can directly enter the spinal cord in young embryos, but access decreases over ~12 h and disappears soon after hatching. Electrical recordings from central Rohon-Beard axons in young embryos showed firing in response to skin impulses. However, unit recordings from Rohon-Beard somata showed that individuals that responded to touch within a characteristic, localised receptive field did not fire to skin impulses, whereas others from similar locations responded reliably. Developmental loss of skin impulse access to the spinal cord mirrored the known spread of sensitivity to gentle touch as the peripheral mechanosensory endings of Rohon-Beard neurons mature. Together, these results suggest that Rohon-Beard neurons respond to skin impulses only while immature, providing a transitory route for skin impulses to excite the CNS. In this way, Rohon-Beard neurons would mediate responses first to noxious and then to localised, gentle touch stimuli as the neurons developed.


Synergy is documented in vitro with low-dose recombinant tumor necrosis factor, cisplatin, and doxorubicin in ovarian cancer cells.
Ovarian carcinomas have been shown to be sensitive or insensitive to the in vitro exposure of several cytotoxic drugs and cytokines. Because of the potential for cytokines to enhance the efficacy of chemotherapeutic agents and to improve their therapeutic index, the optimal dose and schedule of the combination of these agents have been studied. We examined the cytotoxic effect of a combined modality using a variety of concentrations of recombinant tumor necrosis factor (rTNF) (a cytotoxic cytokine) with Adriamycin (ADR) and cisplatin (CDDP) on human ovarian carcinoma cell lines. Cytotoxicity was determined in a 24-hr 51Cr-release assay and confirmed in a 5-day viability culture assay. Five cell lines were used: PA-1, 222, OVCAR-3, SKOV-3, and OVCAR-8. Doses of rTNF that were minimally cytotoxic resulted in significant cytotoxicity and synergy when used with optimal or suboptimal concentrations of ADR or CDDP. This synergy was observed in four cell lines. Interestingly, the rTNF- and drug resistant SKOV-3 cell line was sensitive to the synergistic effect of Adriamycin and rTNF. The synergistic effect that was obtained was specific to rTNF, while the combined use of ADR and CDDP or recombinant interleukin-2 and cytotoxic drugs had no synergistic effect on tumor cell lysis. Further, the addition of anti-TNF antibody abrogated the synergistic effect seen with rTNF and the cytotoxic drugs. These studies demonstrate clearly that significant synergistic antitumor cytotoxic activity against human ovarian carcinoma cell lines can be achieved with combinations of low doses of rTNF and ADR or CDDP, suggesting their possible adaptation in vivo for cancer therapy. Further, the studies suggest that rTNF and the cytotoxic drugs tested may share a common lytic pathway. Since rTNF used alone has been relatively inactive in clinical trials, its potential activity may be apparent only when combined with conventional cytotoxic chemotherapeutic agents and when administered in relatively low concentration.


[Narcolepsy with cataplexy].
Narcolepsy is a rare, disabling sleep disorder, with a prevalence of 20 to 30 per 100,000. Its onset, from childhood to the fifties, peaks in the second decade. The main features are excessive daytime sleepiness and cataplexy or sudden loss of muscle tone triggered by emotional situations. Other less consistent symptoms include hypnagogic hallucinations, sleep paralysis, sleep maintenance insomnia, REM sleep behavior disorders, attention deficit and weight gain at disease onset. Narcolepsy with cataplexy remains a clinical diagnosis but nighttime and daytime polysomnography (multiple sleep latency tests) are useful to document a mean sleep latency below 8 min and at least two sleep-onset REM periods. HLA typing shows an association with HLA DQB1*0602 in more than 92% of cases but was not included in the new diagnostic criteria. In contrast, a low hypocretin levels (values below 110 pg/ml) in the cerebrospinal fluid (CSF) was highly specific for narcolepsy with cataplexy. The deficiency of the hypocretin system is well-established in animal models of narcolepsy (murine and canine narcolepsy) but also in human narcoleptics with a 90% reduction of CSF hypocretin levels in relation with an early loss of hypocretin neurons. The cause of human narcolepsy remains unknown, however an autoimmune process is most probable. The treatment of narcolepsy includes stimulants against sleepiness (modafinil, methylphenidate), anticataplectic drugs (antidepressants) and sodium oxybate. The current therapeutic target is oriented towards hypocretine agonists, histamine (an arousal system) H3 antagonists and immunosuppressants.


Ventricular tachycardia during arthroscopic shoulder surgery: a report of two cases.
We routinely have performed arthroscopic shoulder surgery under general anesthesia in the beach chair position using epinephrine (0.33 mg/L) saline irrigation. At a 2-week interval, two patients, a 19-year-old man scheduled to undergo an arthroscopic Bankart repair for left traumatic anterior instability and a 49-year-old woman scheduled for an arthroscopic rotator cuff repair for a left rotator cuff tear, were resuscitated by chest compression and defibrillation due to a sudden developed cardiogenic shock following ventricular tachycardia at the time of arthroscopic shoulder surgery. They were transferred to the intensive care unit because their emergent echocardiogram showed significantly decreased cardiac functions. They were fully recovered and then discharged. Epinephrine was considered to be the cause of ventricular tachycardia because the two patients showed no anaphylactic reaction to drugs or symptoms of air embolism related to the beach chair position. In addition, according to our observation of epinephrine flow patterns, it was more likely that highly concentrated epinephrine was rapidly infused into the body. This complication is very rare. However, thorough understanding of the side effects and their development of epinephrine during arthroscopic shoulder surgery should neither be overemphasized nor disregarded.


Weight and prognosis for influenza A(H1N1)pdm09 infection during the pandemic period between 2009 and 2011: a systematic review of observational studies with meta-analysis.
In 2009, a novel influenza A(H1N1)pdm09 virus was detected and caused influenza pandemic. It is important to identify the risk factors for severe outcomes. However, inconsistent results regarding the effect of obesity were reported in previous studies. We conducted a systematic review to assess the association between obesity and poor prognosis for laboratory-confirmed A(H1N1)pdm09 influenza. We searched three English databases and three Chinese databases for relevant studies from April 2009 to October 2015: PubMed, the Cochrane library, Embase, CNKI, CBM, and Wanfang. Two investigators independently identified eligible articles, assessed quality using NOS, and extracted data. We performed meta-analyses and meta-regressions to estimate the association between weight and poor prognosis for influenza A(H1N1)pdm09 infection, when data were available. We identified 22 articles enrolling 25,189 laboratory confirmed patients. The pooled estimates indicated obesity significantly increased the risk of fatal and critical complications of influenza A(H1N1)pdm09 infection (for fatal, OR = 1.81, 95% CI: 1.23-2.65; for critical complications, OR = 1.67, 95% CI: 1.13-2.47). However, we found significant interaction between early antiviral treatment and obesity (β = -0.28). After adjustment for early antiviral treatment, relationship between obesity and poor outcomes disappeared (OR = 1.14, 95% CI: 0.94-1.39). The results of the meta-analyses showed obesity significantly increased the risk of death, critical complications, and severe complications for influenza A(H1N1)pdm09 infection, especially among high-quality studies and in Asia region. Importantly, the result from our meta-regression indicated that the conclusion should be interpreted with caution, because early antiviral treatment might be a key confounding factor.


22q11 Gene dosage establishes an adaptive range for sonic hedgehog and retinoic acid signaling during early development.
We asked whether key morphogenetic signaling pathways interact with 22q11 gene dosage to modulate the severity of cranial or cardiac anomalies in DiGeorge/22q1 deletion syndrome (22q11DS). Sonic hedgehog (Shh) and retinoic acid (RA) signaling is altered in the brain and heart-clinically significant 22q11DS phenotypic sites-in LgDel mouse embryos, an established 22q11DS model. LgDel embryos treated with cyclopamine, an Shh inhibitor, or carrying mutations in Gli3(Xtj), an Shh-signaling effector, have morphogenetic anomalies that are either not seen, or seen at significantly lower frequencies in control or single-mutant embryos. Similarly, RA exposure or genetic loss of RA function via heterozygous mutation of the RA synthetic enzyme Raldh2 induces novel cranial anomalies and enhances cardiovascular phenotypes in LgDel but not other genotypes. These changes are not seen in heterozygous Tbx1 mutant embryos-a 22q11 gene thought to explain much of 22q11DS pathogenesis-in which Shh or RA signaling has been similarly modified. Our results suggest that full dosage of 22q11 genes beyond Tbx1 establish an adaptive range for morphogenetic signaling via Shh and RA. When this adaptive range is constricted by diminished dosage of 22q11 genes, embryos are sensitized to otherwise benign changes in Shh and RA signaling. Such sensitization, in the face of environmental or genetic factors that modify Shh or RA signaling, may explain variability in 22q11DS morphogenetic phenotypes.


In situ analysis of alpha-adrenoceptors on arteriolar and venular smooth muscle in rat skeletal muscle microcirculation.
The purpose of this study was to determine whether both alpha 1- and alpha 2-adrenergic receptors exist on vascular smooth muscle of microvessels and whether adrenergic constriction of anatomically distinct microvascular segments is differentially subserved by either receptor subtype. The cremaster skeletal muscle of anesthetized rats was acutely denervated and suspended in a Krebs bath containing cocaine, normetanephrine, and propranolol to block uptake1, uptake2, and beta-receptors, respectively. Intravital microscopy was used to study large distributing arterioles (mean diameter, 100 microns), small precapillary arterioles (25 microns), and capacitance venules (140 microns). Concentration-response (diameter change) curves were obtained for bath-added agonists norepinephrine (mixed alpha 1/alpha 2), phenylephrine (alpha 1), and B-HT 933 (alpha 2) in the absence or presence of antagonists prazosin (alpha 1) and yohimbine (alpha 2). Apparent pD2(-log ED50) values for large arterioles and venules were, respectively, as follows: norepinephrine (7.41 and 7.15), phenylephrine (5.95 and 5.41), and B-HT 933 (5.05 and 5.06). Low concentrations of prazosin (10(-8) M) and yohimbine (10(-7) M) produced receptor subtype-selective antagonism and parallel, dextral displacement of norepinephrine curves for large arterioles and venules. The large arteriole pKB (-log KB) was 7.83 +/- 0.65 for prazosin and 7.36 +/- 0.46 for yohimbine. Higher concentrations of prazosin (10(-7) and 3 X 10(-7) M) and yohimbine (10(-6) M) produced further dextral but nonparallel displacement of norepinephrine curves. In contrast, receptor subtype-selective concentrations of only yohimbine inhibited adrenergic constriction of small, precapillary arterioles; but prazosin had no effect at receptor subtype-selective concentrations. These data suggest that adrenergic regulation of large arterioles and venules in skeletal muscle uses both alpha 1- and alpha 2-adrenoceptors. Precapillary arterioles, however, may be subserved predominantly by alpha 2-receptors.


Cross-Enterprise Communication and Data Exchange in Radiology in Austria: Technology and Use Cases.
The research project REPO (Radiology Ehealth PlatfOrm) was started in 2017 with the goal to "enable cross-enterprise collaboration in radiology using the Austrian eHealth infrastructure". The objective of this paper was to provide an overview of the radiology IT environment - actors, use cases and technology. We conducted semi-structured expert interviews with radiologists and hospital operators and we statistically analyzed the client database of our research project partner. Interviews led to a list of use cases where cross-enterprise collaboration in radiology takes place and the data analysis provided insights on the systems, networks and standards in place. The Austrian IT infrastructure in radiology is a heterogeneous naturally grown environment. Future developments should be based on internationally accorded standards and on integration profiles provided by Integrating the Healthcare Enterprise (IHE).


Barcoding reveals complex clonal dynamics of de novo transformed human mammary cells.
Most human breast cancers have diversified genomically and biologically by the time they become clinically evident. Early events involved in their genesis and the cellular context in which these events occur have thus been difficult to characterize. Here we present the first formal evidence of the shared and independent ability of basal cells and luminal progenitors, isolated from normal human mammary tissue and transduced with a single oncogene (KRAS(G12D)), to produce serially transplantable, polyclonal, invasive ductal carcinomas within 8 weeks of being introduced either subrenally or subcutaneously into immunodeficient mice. DNA barcoding of the initial cells revealed a dramatic change in the numbers and sizes of clones generated from them within 2 weeks, and the first appearance of many 'new' clones in tumours passaged into secondary recipients. Both primary and secondary tumours were phenotypically heterogeneous and primary tumours were categorized transcriptionally as 'normal-like'. This system challenges previous concepts that carcinogenesis in normal human epithelia is necessarily a slow process requiring the acquisition of multiple driver mutations. It also presents the first description of initial events that accompany the genesis and evolution of malignant human mammary cell populations, thereby contributing new understanding of the rapidity with which heterogeneity in their properties can develop.


Management of acute aortic syndromes.
Acute aortic syndrome (AAS) is a modern term to describe interrelated emergency aortic conditions with similar clinical characteristics and challenges. These conditions include aortic dissection, intramural haematoma (IMH), and penetrating atherosclerotic ulcer (PAU and aortic rupture); trauma to the aorta with intimal laceration may also be considered. The common denominator of AAS is disruption of the media layer of the aorta with bleeding within IMH, along the aortic media resulting in separation of the layers of the aorta (dissection), or transmurally through the wall in the case of ruptured PAU or trauma. Population-based studies suggest that the incidence of acute dissection ranges from 2 to 3.5 cases per 100 000 person-years; hypertension and a variety of genetic disorders with altered connective tissues are the most prevalent risk conditions. Patients with AAS often present in a similar fashion, regardless of the underlying condition of dissection, IMH, PAU, or contained aortic rupture. Pain is the most commonly presenting symptom of acute aortic dissection and should prompt immediate attention including diagnostic imaging modalities (such as multislice computed tomography, transoesophageal ultrasound, or magnetic resonance imaging). Prognosis is clearly related to undelayed diagnosis and appropriate surgical repair in the case of proximal involvement of the aorta; affection of distal segments of the aorta may call for individualized therapeutic approaches favouring endovascular in the presence of malperfusion or imminent rupture, or medical management.


Estimating the impact of mandatory fortification of bread with iodine on pregnant and post-partum women.
Iodine deficiency has re-emerged in Australia. Pregnant and breastfeeding women need higher iodine intakes (estimated average requirements: 160 μg/day and 190 μg/day) than non-pregnant women (100 μg/day) because iodine is critical for early infant development. The impact of iodine fortification of bread on women's iodine intake is evaluated by reproductive status using 2003 Australian Longitudinal Study on Women's Health (ALSWH) food frequency data and projected onto 1995 National Nutrition Survey (NNS) daily food consumption data for women of child-bearing age. Recent iodine analyses of Australian foods were combined with reported intakes of key foods to estimate iodine intake before and after fortification for 665 pregnant, 432 zero to 6 months postpartum, 467 seven to 12 months postpartum and 7324 non-pregnant women. Differences in mean iodine intake between these groups were projected onto NNS estimates of total iodine intake for women of child-bearing age. Pregnant and postpartum women reported eating more bread than did non-pregnant women. Mean iodine intakes (μg/day before; and after fortification) from key foods were higher in pregnant (78; 124), 0-6 months postpartum (75; 123) and 7-12 months postpartum (71; 117) than in non-pregnant women (65; 103). Projecting ALSWH results onto the NNS yields total mean iodine intakes of 167, 167, 160 and 146 for the same groups. Current iodine intakes are well below dietary recommendations. The impact of iodine fortification of bread would be greater for pregnant and postpartum women than has been previously estimated using general population intakes, but additional strategies to increase intakes by these groups are still needed.


Oxidative stress antagonizes fluoroquinolone drug sensitivity via the SoxR-SUF Fe-S cluster homeostatic axis.
The level of antibiotic resistance exhibited by bacteria can vary as a function of environmental conditions. Here, we report that phenazine-methosulfate (PMS), a redox-cycling compound (RCC) enhances resistance to fluoroquinolone (FQ) norfloxacin. Genetic analysis showed that E. coli adapts to PMS stress by making Fe-S clusters with the SUF machinery instead of the ISC one. Based upon phenotypic analysis of soxR, acrA, and micF mutants, we showed that PMS antagonizes fluoroquinolone toxicity by SoxR-mediated up-regulation of the AcrAB drug efflux pump. Subsequently, we showed that despite the fact that SoxR could receive its cluster from either ISC or SUF, only SUF is able to sustain efficient SoxR maturation under exposure to prolonged PMS period or high PMS concentrations. This study furthers the idea that Fe-S cluster homeostasis acts as a sensor of environmental conditions, and because its broad influence on cell metabolism, modifies the antibiotic resistance profile of E. coli.


Outcome of hemiepiphyseal stapling for late-onset tibia vara.
The results of hemiepiphysiodesis for the treatment of late-onset tibia vara have been reported to be favorable, but the technique requires careful timing and an accurate estimation of skeletal age. Hemiepiphyseal stapling does not require a careful estimation of skeletal age, and it has been reported to yield good results with low morbidity. However, we are not aware of any study evaluating the intermediate-term radiographic results or complications of this procedure. Twenty-six patients with thirty-three extremities with late-onset tibia vara were treated with proximal tibial hemiepiphyseal stapling. Fourteen extremities had substantial concomitant distal femoral varus and also had hemiepiphyseal stapling of the distal part of the femur. Eighteen patients (twenty-three involved extremities) had juvenileonset tibia vara and eight patients (ten involved extremities) had adolescent-onset tibia vara. The mean age at the time of stapling was 11.8 years. The mean duration of follow-up was 3.8 years. We reviewed standardized standing radiographs to determine the mechanical axis deviation, the medial proximal tibial angle, the lateral distal femoral angle, and the zone of the knee through which the mechanical axis passed. The mean mechanical axis deviation improved from 58 mm (range, 27 to 157 mm) preoperatively to 22 mm (range, -33 to 117 mm) at the time of the last follow-up, and the mean medial proximal tibial angle improved from 77 degrees (range, 50 degrees to 85 degrees ) to 85 degrees (range, 48 degrees to 95 degrees ). In the fourteen lower extremities in which distal femoral hemiepiphyseal stapling was performed, the mean lateral distal femoral angle improved from 96 degrees (range, 92 degrees to 100 degrees ) to 86 degrees (range, 79 degrees to 97 degrees ). At the time of the final follow-up, seven extremities were considered to be in moderate varus; four, in mild varus; twenty, in normal alignment; and two, in valgus. No differences in radiographic outcome were noted between the juvenile and adolescent forms of tibia vara. Only one of the four extremities with severe preoperative varus was corrected to normal alignment; the remaining three were left with moderate varus. Hemiepiphyseal stapling of the lateral aspect of the proximal tibial physis and, as needed, the lateral aspect of the distal femoral physis is safe and effective in children with late-onset tibia vara if the physes are sufficiently open and the varus deformity is mild to moderate. Hemiepiphyseal stapling is particularly effective in patients who are ten years of age or younger. Therapeutic Level IV.


Red cell transfusion management for patients undergoing cardiac surgery for congenital heart disease.
Congenital heart disease is the most commonly diagnosed neonatal congenital condition. Without surgery, only 30% to 40% of patients affected will survive to 10 years old. Mortality has fallen since the 1990s with 2006 to 2007 figures showing surgical survival at one year of 95%. Patients with congenital heart disease are potentially exposed to red cell transfusion at many points in the surgical pathway. There are a number of risks associated with red cell transfusion that may be translated into increased patient morbidity and mortality. To evaluate the effects of red cell transfusion on mortality and morbidity on patients with congenital heart disease at the time of cardiac surgery. We searched 11 bibliographic databases and three ongoing trials databases including the Cochrane Central Register of Controlled Trials (CENTRAL) (Issue 5, 2013), MEDLINE (Ovid, 1950 to 11 June 2013), EMBASE (Ovid, 1980 to 11 June 2013), ClinicalTrials.gov, World Health Organization (WHO) ICTRP and the ISRCTN Register (to June 2013). We also searched references of all identified trials, relevant review articles and abstracts from between 2006 and 2010 of the most relevant conferences. We did not limit the searches by language of publication. We included randomised controlled trials (RCTs) comparing red cell transfusion interventions in patients undergoing cardiac surgery for congenital heart disease. We included participants of any age (neonates, paediatrics and adults) and with any type of congenital heart disease (cyanotic or acyanotic). We excluded patients with congenital heart disease undergoing non-cardiac surgery. No co-morbidities were excluded. Two review authors independently assessed trial quality and extracted data. We contacted study authors for additional information. We identified 11 trials (862 participants). All trials were in neonatal or paediatric populations. The trials covered only three areas of interest: restrictive versus liberal transfusion triggers (two trials), leukoreduction versus non-leukoreduction (two trials) and standard versus non-standard cardiopulmonary bypass (CPB) prime (seven trials). Owing to the clinical diversity in the participant groups (cyanotic (three trials), acyanotic (four trials) or mixed (four trials)) and the intervention groups, it was not appropriate to pool data in a meta-analysis. No study reported data for all the outcomes of interest to this review. Risk of bias was mixed across the included trials, with only attrition bias being low across all trials. Blinding of study personnel and participants was not always possible, depending on the intervention being used.Five trials (628 participants) reported the primary outcome: 30-day mortality. In three trials (a trial evaluating restrictive and liberal transfusion (125 participants), a trial of cell salvage during CPB (309 participants) and a trial of washed red blood cells during CPB (128 participants)), there was no clear difference in mortality at 30 days between the intervention arms. In two trials comparing standard and non-standard CPB prime, there were no deaths in either randomised group. Long-term mortality was similar between randomised groups in one trial each comparing restrictive and liberal transfusion or standard and non-standard CPB prime.Four trials explored a range of adverse effects following red cell transfusion. Kidney failure was the only adverse event that was significantly different: patients receiving cell salvaged red blood cells during CPB were less likely to have renal failure than patients not exposed to cell salvage (risk ratio (RR) 0.26, 95% confidence interval (CI) 0.09 to 0.79, 1 study, 309 participants). There was insufficient evidence to determine whether there was a difference between transfusion strategies for any other severe adverse events.The duration of mechanical ventilation was measured in seven trials (768 participants). Overall, there was no consistent difference in the duration of mechanical ventilation between the intervention and control arms.The duration of intensive care unit (ICU) stay was measured in six trials (459 participants). There was no clear difference in the duration of ICU stay between the intervention arms in the transfusion trigger and leukoreduction trials. In the standard versus non-standard CPB prime trials, one trial examining the impact of washing transfused bypass prime red blood cells showed no clear difference in duration of ICU stay between the intervention arms, while the trial assessing ultrafiltration of the priming blood showed a shorter duration of ICU stay in the ultrafiltration group. There are only a small number of small and heterogeneous trials so there is insufficient evidence to assess the impact of red cell transfusion on patients with congenital heart disease undergoing cardiac surgery accurately. It is possible that the presence or absence of cyanosis impacts on trial outcomes, which would necessitate different clinical management of two groups. Further adequately powered, specific, high-quality trials are warranted to assess this fully.


Interference with the germination and growth of Ulva zoospores by quorum-sensing molecules from Ulva-associated epiphytic bacteria.
Ulva zoospores preferentially settle on N-acylhomoserine lactone (AHL) producing marine bacterial biofilms. To investigate whether AHL signal molecules also affect the success and rate of zoospore germination in addition to zoospore attraction, the epiphytic bacteria associated with mature Ulva linza were characterized and bacterial isolates representative of this community tested for the ability to produce AHLs. Two of these AHL-producing isolates, Sulfitobacter spp. 376 and Shewanella spp. 79, were transformed with plasmids expressing the Bacillus spp. AHL lactonase gene aiiA to generate AHL-deficient variants. The germination and growth of U. linza zoospores was studied in the presence of these AHL-deficient strains and their AHL-producing counterparts. This revealed that the AHLs produced by Sulfitobacter spp. and Shewanella spp. or the bacterial products they regulate have a negative impact on both zoospore germination and the early growth of the Ulva germling. Further experiments with Escherichia coli biofilms expressing recombinant AHL synthases and synthetic AHLs provide data to demonstrate that zoospores germinated and grown in the absence of AHLs were significantly longer than those germinated in the presence of AHLs. These results reveal an additional role for AHLs per se in the interactive relationships between marine bacteria and Ulva zoospores.


Parent-of-origin-specific allelic associations among 106 genomic loci for age at menarche.
Age at menarche is a marker of timing of puberty in females. It varies widely between individuals, is a heritable trait and is associated with risks for obesity, type 2 diabetes, cardiovascular disease, breast cancer and all-cause mortality. Studies of rare human disorders of puberty and animal models point to a complex hypothalamic-pituitary-hormonal regulation, but the mechanisms that determine pubertal timing and underlie its links to disease risk remain unclear. Here, using genome-wide and custom-genotyping arrays in up to 182,416 women of European descent from 57 studies, we found robust evidence (P < 5 × 10(-8)) for 123 signals at 106 genomic loci associated with age at menarche. Many loci were associated with other pubertal traits in both sexes, and there was substantial overlap with genes implicated in body mass index and various diseases, including rare disorders of puberty. Menarche signals were enriched in imprinted regions, with three loci (DLK1-WDR25, MKRN3-MAGEL2 and KCNK9) demonstrating parent-of-origin-specific associations concordant with known parental expression patterns. Pathway analyses implicated nuclear hormone receptors, particularly retinoic acid and γ-aminobutyric acid-B2 receptor signalling, among novel mechanisms that regulate pubertal timing in humans. Our findings suggest a genetic architecture involving at least hundreds of common variants in the coordinated timing of the pubertal transition.


Hypertrehalosaemic neuropeptides decrease levels of the glycolytic signal fructose 2,6-bisphosphate in cockroach fat body
In cockroach fat body, trehalogenesis and glycolysis compete for glucose phosphates as common substrates. During trehalogenesis, glycolysis is inhibited, although the mechanism responsible for this is not known. Incubation of the isolated fat body from the Argentine cockroach Blaptica dubia with an extract of the corpora cardiaca containing as little as 0.005 gland equivalents ml-1 of incubation medium increased the release of trehalose (anthrone-positive material) from the tissue by more than 100 %. The content of the glycolytic activator fructose 2,6-bisphosphate was decreased by up to 50 %. A decapeptide was isolated from the corpora cardiaca of B. dubia and shown to be identical to the naturally occurring Blaberus discoidalis hypertrehalosaemic peptide (Bld HrTH), which was also found in the corpora cardiaca. Synthetic Bld HrTH at 2 nmol l-1 and above increased trehalose production and decreased the content of fructose 2,6-bisphosphate to the same extent as did corpus cardiacum extract. The octapeptides Periplaneta americana cardioacceleratory hormones I and II (Pea CAH-I and Pea CAH-II) also had a significant effect on both parameters. Fructose 2,6-bisphosphate is a potent activator of phosphofructokinase from cockroach fat body if the enzyme is assayed at near-physiological concentrations of substrates and effectors. It is suggested that, because of the decrease in fructose 2,6-bisphosphate levels in the fat body, the activity of the key glycolytic enzyme phosphofructokinase is diminished. This can explain the inhibition of glycolytic flux by hypertrehalosaemic peptides which alters the balance of glucose metabolism in favour of trehalose formation.


Stochastic resetting by a random amplitude.
Stochastic resetting, a diffusive process whose amplitude is reset to the origin at random times, is a vividly studied strategy to optimize encounter dynamics, e.g., in chemical reactions. Here we generalize the resetting step by introducing a random resetting amplitude such that the diffusing particle may be only partially reset towards the trajectory origin or even overshoot the origin in a resetting step. We introduce different scenarios for the random-amplitude stochastic resetting process and discuss the resulting dynamics. Direct applications are geophysical layering (stratigraphy) and population dynamics or financial markets, as well as generic search processes.


Imperfect genes, Fisherian mutation and the evolution of sex.
In this paper we present a mathematical model of mutation and selection that allows for the coexistence of multiple alleles at a locus with very small selective differences between alleles. The model also allows for the determination of fitness by multiple loci. Models of this sort are biologically plausible. However, some previous attempts to construct similar models have assumed that all mutations produce a decrease in fitness, and this has led to a tendency for the average fitness of population members to decline when population numbers are finite. In our model we incorporate some of the ideas of R. A. FISHER, so that both deleterious and beneficial mutations are possible. As a result, average fitness tends to approach a stationary distribution. We have used computer simulation methods to apply the Fisherian mutation model to the problem of the evolution of sex and recombination. The results suggest that sex and recombination can provide very large benefits in terms of average fitness. The results also suggest that obligately sexual species will win ecological competitions with species that produce a substantial fraction of their offspring asexually, so long as the number of sites under selection within the genomes of the competing species is not too small and the population sizes are not too large. Our model focuses on fertility selection in an hermaphroditic plant. However, the results are likely to generalize to a wide variety of other situations as well.