Spatial and management factors associated with exposure of smallholder dairy cattle in Tanzania to tick-borne pathogens.
A cross-sectional study of serum antibody responses of cattle to tick-borne pathogens (Theileria parva, Theileria mutans,Anaplasma marginale, Babesia bigemina and Babesia bovis) was conducted on smallholder dairy farms in Tanga and Iringa Regions of Tanzania. Seroprevalence was highest for T. parva (48% in Iringa and 23% in Tanga) and B. bigemina (43% in Iringa and 27% in Tanga) and lowest for B. bovis (12% in Iringa and 6% in Tanga). We use spatial and non-spatial models, fitted using classical and Bayesian methods, to explore risk factors associated with seroprevalence. These include both fixed effects (age, grazing history and breeding status) and random effects (farm and local spatial effects). In both regions, seroprevalence for all tick-borne pathogens increased significantly with age. Animals pasture grazed in the 3 months prior to the start of the sampling period were significantly more likely to be seropositive for Theileria spp. and Babesia spp. Pasture grazed animals were more likely to be seropositive than zero-grazed animals for A. marginale, but the relationship was weaker than that observed for the other four pathogens. This study did not detect any significant differences in seroprevalence associated with other management-related variables, including the method or frequency of acaricide application. After adjusting for age, there was weak evidence of localised (<5 km) spatial correlation in exposure to some of the tick borne diseases. However, this was small compared with the 'farm-effect', suggesting that risk factors specific to the farm were more important than those common to the local neighbourhood. Many animals were seropositive for more than one pathogen and the correlation between exposure to the different pathogens remained after adjusting for the identified risk factors. Identifying the determinants of exposure to multiple tick-borne pathogens and characterizing local variation in risk will assist in the development of more effective control strategies for smallholder dairy farms.


Mitochondrial heat shock protein machinery hsp70/hsp40 is indispensable for proper mitochondrial DNA maintenance and replication.
Mitochondrial chaperones have multiple functions that are essential for proper functioning of mitochondria. In the human-pathogenic protist Trypanosoma brucei, we demonstrate a novel function of the highly conserved machinery composed of mitochondrial heat shock proteins 70 and 40 (mtHsp70/mtHsp40) and the ATP exchange factor Mge1. The mitochondrial DNA of T. brucei, also known as kinetoplast DNA (kDNA), is represented by a single catenated network composed of thousands of minicircles and dozens of maxicircles packed into an electron-dense kDNA disk. The chaperones mtHsp70 and mtHsp40 and their cofactor Mge1 are uniformly distributed throughout the single mitochondrial network and are all essential for the parasite. Following RNA interference (RNAi)-mediated depletion of each of these proteins, the kDNA network shrinks and eventually disappears. Ultrastructural analysis of cells depleted for mtHsp70 or mtHsp40 revealed that the otherwise compact kDNA network becomes severely compromised, a consequence of decreased maxicircle and minicircle copy numbers. Moreover, we show that the replication of minicircles is impaired, although the lack of these proteins has a bigger impact on the less abundant maxicircles. We provide additional evidence that these chaperones are indispensable for the maintenance and replication of kDNA, in addition to their already known functions in Fe-S cluster synthesis and protein import. Impairment or loss of mitochondrial DNA is associated with mitochondrial dysfunction and a wide range of neural, muscular, and other diseases. We present the first evidence showing that the entire mtHsp70/mtHsp40 machinery plays an important role in mitochondrial DNA replication and maintenance, a function likely retained from prokaryotes. These abundant, ubiquitous, and multifunctional chaperones share phenotypes with enzymes engaged in the initial stages of replication of the mitochondrial DNA in T. brucei.


Cardiac pacing in children.
Advances in pacemaker technology over the last 25 years have made cardiac pacing in infants and children a safe and practical therapy. Some of the technical challenges encountered with the first permanent pacemaker implantation in children during the early 1960s have been solved with miniaturized generators, lithium batteries, noninvasive programmability, and improved placement techniques. This chapter reports common causes of bradydysrhythmias in children, reviews the indications for permanent pacemaker implantation in children, describes current pacing systems appropriate for children, and identifies nursing implications and potential pacemaker complications unique to children.


The role of platelets and megakaryocytes in bone metastasis.
Blood platelets have been known for more than a century as important partners for successful metastatic dissemination of solid tumors. Cancer cell-induced platelet activation is a key event responsible for prometastatic activity of platelets. Blocking platelet aggregation inhibits the progression of skeletal metastases through mechanisms that are not fully understood. The establishment and progression of bone metastases are strongly influenced by the bone remodeling process. Growth factors and cytokines released upon platelet activation may contribute to both skeletal tumor growth and osteolytic lesions. Megakaryocytes are platelet precursors located in the bone marrow that control bone mass through direct stimulation of osteoblast functions and indirect inhibition of osteoclast activities. Considering growing evidence for their role in the metastatic cascade, platelets and/or megakaryocytes may provide new therapeutic opportunities to help limit bone metastases.


Indeterminacy in brain and behavior.
The central goal of modern science that evolved during the Enlightenment was the empirical reduction of uncertainty by experimental inquiry. Although there have been challenges to this view in the physical sciences, where profoundly indeterminate events have been identified at the quantum level, the presumption that physical phenomena are fundamentally determinate seems to have defined modern behavioral science. Programs like those of the classical behaviorists, for example, were explicitly anchored to a fully deterministic worldview, and this anchoring clearly influenced the experiments that those scientists chose to perform. Recent advances in the psychological, social, and neural sciences, however, have caused a number of scholars to begin to question the assumption that all of behavior can be regarded as fundamentally deterministic in character. Although it is not yet clear whether the generative mechanisms for human and animal behavior will require a philosophically indeterminate approach, it is clear that behavioral scientists of all kinds are beginning to engage the issues of indeterminacy that plagued physics at the beginning of the twentieth century.


Poly(ionic liquid) composites.
Poly(ionic liquid)s (PILs), as an innovative class of polyelectrolytes, are composed of polymeric backbones with IL species in each repeating unit. The combined merits of the polymers and ILs make them promising materials for composites in materials science. Particularly, the integration of PILs with functional substances (PIL composites) opens up a new dimension in utilizing ionic polymers by offering novel properties and improved functions, which impacts multiple subfields of our chemical society. This review summarizes recent developments of PIL composites with a special emphasis on the preparation techniques that are based on the intrinsic properties of the PILs and the synergistic effects between the PILs and substances of interest for diverse applications.


Serogroup B meningococcal vaccines-an unfinished story.
Most invasive meningococcal disease in developed countries is caused by Neisseria meningitidis with a serogroup B capsule. However, despite availability of vaccines for other serogroups since the 1960s, no serogroup B vaccine exists. In this Review we look at the development of serogroup B vaccines over the past 40 years. Outer membrane vesicle vaccines have been successfully used to control geographically isolated epidemics, but most have not been highly immunogenic in young children or provided broad cross-protection from infections with other strains. Vaccines based on subcapsular antigens have recently produced promising results in early clinical trials, and the disease burden might be substantially reduced over the next few years.


Ca2+-dependent translocation of the calcyclin-binding protein in neurons and neuroblastoma NB-2a cells.
The calcyclin-binding protein (CacyBP) binds calcyclin (S100A6) at physiological levels of [Ca(2+)] and is highly expressed in brain neurons. Subcellular localization of CacyBP was examined in neurons and neuroblastoma NB-2a cells at different [Ca(2+)](i). Immunostaining indicates that CacyBP is present in the cytoplasm of unstimulated cultured neurons in which resting [Ca(2+)](i) is known to be approximately 50 nm. When [Ca(2+)](i) was increased to above 300 nm by KCl treatment, the immunostaining was mainly apparent as a ring around the nucleus. Such perinuclear localization of CacyBP was observed in untreated neuroblastoma NB-2a cells in which [Ca(2+)](i) is approximately 120 nm. An additional increase in [Ca(2+)](i) to above 300 nm by thapsigargin treatment did not change CacyBP localization. However, when [Ca(2+)](i) in NB-2a cells dropped to 70 nm, because of BAPTA/AM treatment, perinuclear localization was diminished. Ca(2+)-induced translocation of CacyBP was confirmed by immunogold electron microscopy and by fluorescence of NB-2a cells transfected with an EGFP-CacyBP vector. Recombinant CacyBP can be phosphorylated by protein kinase C in vitro. In untreated neuroblastoma NB-2a cells, CacyBP is phosphorylated on a serine residue(s), but exists in the dephosphorylated form in BAPTA/AM-treated cells. Thus, phosphorylation of CacyBP occurs in the same [Ca(2+)](i) range that leads to its perinuclear translocation.


Non-invasive detection of microvascular changes in a paediatric and adolescent population with type 1 diabetes: a pilot cross-sectional study.
The detection of microvascular damage in type 1 diabetes is difficult and traditional investigations do not detect changes until they are well established. The purpose of this study was to investigate the combined ability of nailfold capillaroscopy, laser Doppler flowmetry, retinal vessel analysis and 24-hr ambulatory blood pressure monitoring to detect early microvascular changes in a paediatric and adolescent population with type 1 diabetes. Patients aged between 8 - 18 years with type I diabetes and no other autoimmune conditions were studied. The participants underwent the above cardiac and vascular investigations in a single three-hour session. Standard parameters including HbA1c were also investigated. Associations between all parameters were described by correlation analysis. Fisher's exact and t-tests determined the association with clinical findings. 26 participants were recruited. The mean HbA1c was 8.1% (SD ± 1.1) with a mean duration of type 1 diabetes of 7.9 years (SD ± 3.4). Three participants had microalbuminuria and one had early signs of retinopathy. Participants with microvascular complications had more avascular areas on nailfold capillaroscopy (p = 0.03). Recent HbA1c was positively associated with the number of nailfold microhaemorrhages (p = 0.03) Decreased baseline perfusion by laser Doppler flowmetry was associated with increased capillary density (p = 0.001) and an increased number of microaneurysms (p = 0.04) on nailfold capillaroscopy. This pilot study has shown that in children and adolescents with established type 1 diabetes, abnormal microvasculature can be detected by these investigations. These markers were also positively associated with evidence of suboptimal diabetes control as assessed by HbA1c. Further research will be necessary to determine the practical role of these investigations in the management and progress of the complications of type 1 diabetes. Clinical Trial number NCT01279928, ClinicalTrials.gov.


Mall Walking Program Environments, Features, and Participants: A Scoping Review.
Walking is a preferred and recommended physical activity for middle-aged and older adults, but many barriers exist, including concerns about safety (ie, personal security), falling, and inclement weather. Mall walking programs may overcome these barriers. The purpose of this study was to summarize the evidence on the health-related value of mall walking and mall walking programs. We conducted a scoping review of the literature to determine the features, environments, and benefits of mall walking programs using the RE-AIM framework (reach, effectiveness, adoption, implementation, and maintenance). The inclusion criteria were articles that involved adults aged 45 years or older who walked in indoor or outdoor shopping malls. Exclusion criteria were articles that used malls as laboratory settings or focused on the mechanics of walking. We included published research studies, dissertations, theses, conference abstracts, syntheses, nonresearch articles, theoretical papers, editorials, reports, policy briefs, standards and guidelines, and nonresearch conference abstracts and proposals. Websites and articles written in a language other than English were excluded. We located 254 articles on mall walking; 32 articles met our inclusion criteria. We found that malls provided safe, accessible, and affordable exercise environments for middle-aged and older adults. Programmatic features such as program leaders, blood pressure checks, and warm-up exercises facilitated participation. Individual benefits of mall walking programs included improvements in physical, social, and emotional well-being. Limited transportation to the mall was a barrier to participation. We found the potential for mall walking programs to be implemented in various communities as a health promotion measure. However, the research on mall walking programs is limited and has weak study designs. More rigorous research is needed to define best practices for mall walking programs' reach, effectiveness, adoption, implementation, and maintenance.


Treatment of retroperitoneal sarcoma: current standards and new developments.
Retroperitoneal sarcomas are rare tumors and with complex treatment. In this manuscript we give an overview of current standards in treatment of this disease and discuss new developments. Surgery with complete resection of the primary tumor is still the only curative modality. The role of preoperative radiotherapy is not clear and is currently being investigated in a clinical trial. Neo-adjuvant chemotherapy is not the standard of care but can be considered occasionally when complete resection is uncertain. Local and distant recurrent disease carries a dismal prognosis, although long-term survival can be achieved. Liposarcomas tend to recur locally, whereas distant recurrences are more often seen in leiomyosarcoma and other subtypes. Outcome improves when patients are treated in high volume sarcoma centers. In the metastatic setting, newer systemic agents have recently been approved. Treatment of retroperitoneal sarcomas is complex and all patients should be treated in multidisciplinary sarcoma centers. Increasing international collaboration of expert centers in sharing expertise and performing clinical trials might lead to better treatment and improved survival.


Mapping the potential risk of mycetoma infection in Sudan and South Sudan using ecological niche modeling.
In 2013, the World Health Organization (WHO) recognized mycetoma as one of the neglected tropical conditions due to the efforts of the mycetoma consortium. This same consortium formulated knowledge gaps that require further research. One of these gaps was that very few data are available on the epidemiology and transmission cycle of the causative agents. Previous work suggested a soil-borne or Acacia thorn-prick-mediated origin of mycetoma infections, but no studies have investigated effects of soil type and Acacia geographic distribution on mycetoma case distributions. Here, we map risk of mycetoma infection across Sudan and South Sudan using ecological niche modeling (ENM). For this study, records of mycetoma cases were obtained from the scientific literature and GIDEON; Acacia records were obtained from the Global Biodiversity Information Facility. We developed ENMs based on digital GIS data layers summarizing soil characteristics, land-surface temperature, and greenness indices to provide a rich picture of environmental variation across Sudan and South Sudan. ENMs were calibrated in known endemic districts and transferred countrywide; model results suggested that risk is greatest in an east-west belt across central Sudan. Visualizing ENMs in environmental dimensions, mycetoma occurs under diverse environmental conditions. We compared niches of mycetoma and Acacia trees, and could not reject the null hypothesis of niche similarity. This study revealed contributions of different environmental factors to mycetoma infection risk, identified suitable environments and regions for transmission, signaled a potential mycetoma-Acacia association, and provided steps towards a robust risk map for the disease.


Association between genetic variation in the gene for insulin-like growth factor-I and low birthweight.
Low birthweight is associated with later risk of type 2 diabetes and related disorders. We aimed to show that a polymorphism in the gene for insulin-like growth factor-I, which has proved to raise risk of type 2 diabetes and myocardial infarction, is associated with low birthweight. We recorded birthweight and obtained DNA for 463 adults. Individuals who did not have the wild-type allele of the polymorphism had a 215 g lower birthweight than those homozygous for this allele (95% CI -411 to -10). Our data lend support to the hypothesis that genetic variation affecting fetal growth could account for the association between low birthweight and susceptibility to diabetes and cardiovascular disease in later life.


Testosterone and dihydrotestosterone and incident ischaemic stroke in men in the Cardiovascular Health Study.
Ischaemic stroke is a major cause of morbidity and mortality in elderly men. Our main objective was to examine whether testosterone (T) or dihydrotestosterone (DHT) was associated with incident ischaemic stroke in elderly men. Cohort study. Elderly men in the Cardiovascular Health Study who had no history of stroke, heart disease or prostate cancer as of 1994 and were followed until December 2010. Adjudicated ischaemic stroke. Among 1032 men (mean age 76, range 66-97), followed for a median of 10 years, 114 had an incident ischaemic stroke. Total T and free T were not significantly associated with stroke risk, while DHT had a nonlinear association with incident stroke (P = 0·006) in analyses adjusted for stroke risk factors. The lowest risk of stroke was at DHT levels of 50-75 ng/dl, with greater risk of stroke at DHT levels above 75 ng/dl or below 50 ng/dl. Results were unchanged when SHBG was added to the model. Calculated free DHT had an inverse linear association with incident ischaemic stroke with HR 0·77 (95% CI, 0·61, 0·98) per standard deviation in analyses adjusted for stroke risk factors. Dihydrotestosterone had a nonlinear association with stroke risk in which there was an optimal DHT level associated with the lowest stroke risk. Further studies are needed to confirm these results and to clarify whether there is an optimal androgen range associated with the least risk of adverse outcomes in elderly men.


Proton magnetic resonance spectroscopic imaging of cortical gray and white matter in schizophrenia.
To apply in vivo proton magnetic resonance spectroscopy imaging estimates of N-acetylaspartate (NAA), a neuronal marker, to clarify the relative contribution of neuronal and glial changes to the widespread volume deficit of cortical gray matter seen in patients with schizophrenia with magnetic resonance images. Ten male veterans meeting criteria of the DSM-IV, for schizophrenia and 9 healthy age-matched men for comparison were scanned using spectroscopic, anatomical, and field-map sequences. Instrument and collection variables were standardized to allow an estimation of comparable values for NAA, choline, and creatine for all subjects. Metabolite values from each voxel on 3 upper cortical slices were regressed against the gray tissue proportion of that voxel to derive estimates of gray and white matter NAA, creatine, and choline concentrations. The volume of cortical gray matter was reduced in patients with schizophrenia, but NAA signal intensity from a comparable region was normal. In contrast, the volume of cortical white matter was normal in patients with schizophrenia, but NAA signal intensity from a comparable region was reduced. The lack of reduction in gray matter NAA signal intensity suggests that the cortical gray matter deficit in these patients involved both neuronal and glial compartments rather than a neurodegenerative process in which there is a decrease in the neuronal relative to the glial compartment. Reduced white matter NAA signal intensity without a white matter volume deficit may reflect abnormal axonal connections.


One-pot synthesized functionalized mesoporous silica as a reversed-phase sorbent for solid-phase extraction of endocrine disrupting compounds in milks.
A new procedure for the determination of 12 naturally occurring hormones and some related synthetic chemicals in milk, commonly used as growth promoters in cattle, is reported. The method is based on liquid-liquid extraction followed by solid-phase extraction (SPE) using a new one-pot synthesized ordered mesoporous silica (of the SBA-15 type) functionalized with octadecyl groups (denoted as SBA-15-C18-CO) as reversed-phase sorbent. The analytes were eluted with methanol and then submitted to HPLC with diode array detection. Under optimal conditions, the method quantification limit for the analytes ranged from 0.023 to 1.36μg/mL. The sorbent affored the extraction of estrone, 17β-estradiol, estriol, progesterone, hexestrol, diethylstilbestrol, 4-androstene-3,17-dione, ethinylestradiol, 17α-methyltestosterone, nandrolone, prednisolone and testosterone with mean recoveries ranging from 72% to 105% (except for diethylstilbestrol) with RSD<11%. These results were comparable and, in some cases, even better than those obtained with other extraction methods, therefore SBA-15-C18-CO mesoporous silica possess a high potential as a reversed-phase sorbent for SPE of the 12 mentioned endocrine disrupting compounds in milk samples.


'If your husband calls, you have to go': understanding sexual agency among young married women in urban South India.
Early marriage is common in many developing countries, including India. Women who marry early have little power within their marriage, particularly in the sexual domain. Research is limited on women's ability to control their marital sexual experiences. We identified factors affecting sexual communication among married women aged 16-25, in Bangalore, India, and how factors associated with sexual communication differed from those influencing non-sexual agency. We ran ordered logit regression models for one outcome of sexual agency (sexual communication, n = 735) and two outcomes of non-sexual agency (fertility control, n = 735, and financial decision-making, n = 728). Sexual communication was more restricted (83 women (11.3%) with high sexual communication) than financial decision-making (183 women (25.1%) with high financial decision-making agency) and fertility control (238 women (32.4%) with high fertility control). Feeling prepared before the first sexual experience was significantly associated with sexual communication (odds ratio (OR) = 1.8; 95% confidence interval (CI) = 1.13-2.89). Longer marriage duration (OR 2.13; 95% CI = 1.42-3.20) and having worked pre-marriage (OR 1.38; 95% CI = 1.02-1.86) were also significant. Few other measures of women's resources increased their odds of sexual communication. Education, having children, pre-marital vocational training and marital intimacy were significant for non-sexual outcomes but not sexual communication. Policy-makers seeking to enhance young married women's sexual communication need to consider providing sex education to young women before they marry. More broadly, interventions designed to increase women's agency need to be tailored to the type of agency being examined.


The impact of nonautogenous bone graft on orthodontic treatment.
Guided bone regeneration with alloplastic bone graft substitutes has a high impact in surgical dentistry nowadays. In the future, the orthodontist will inevitably be confronted with patients with such a medical history. A review of the literature was performed to investigate present materials used, indications, and concerns and problems in interdisciplinary treatment of patients with nonautogenous bone grafts. An electronic key word search in the literature database PubMed of the US National Library of Medicine and The Cochrane Library (CENTRAL) of the Cochrane Collaboration was performed. Numerous journals were searched manually, and the reference lists of all identified papers related to the topic were reviewed to warrant completeness. All abstracts of the identified publications were screened in duplicate by two autonomous reviewers. Inclusion criteria were the use of a nonautogenous bone graft followed by orthodontic tooth movement or a tooth eruption through the grafting material. The total number of identified publications was 37. It was possible to divide these into two major topics, orthodontic tooth movement in bone graft and tooth eruption through bone graft. Besides experimental setups, indications were periodontal defects, cystic lesions, and alveolar clefts. Here, different types of bone graft were used. The following tooth movement was partially uneventful and sometimes attended by serious adverse effects. The resulting articles presented a low level of evidence and showed diversity in question, methodology, and type of publication. For this reason, it was not possible to perform a quantifying review. Therefore, the articles are discussed in a descriptive manner with regard to the topic in question. Both orthodontic tooth movement and tooth eruption through grafted bone are not only possible but also might provide promising results. However, more research on this interdisciplinary treatment is necessary to avoid risks and achieve more safety in orthodontic treatment of these patients.


Loss of ASRGL1 expression is an independent biomarker for disease-specific survival in endometrioid endometrial carcinoma.
For endometrial carcinoma, prognostic stratification methods do not satisfactorily identify patients with adverse outcome. Currently, histology, tumor grade and stage are used to tailoring surgical treatment and to determine the need for adjuvant treatment. Low-risk patients are not considered to require adjuvant therapy or staging lymphadenectomy. For patients with intermediate or high risk, some guidelines recommend tailoring adjuvant treatment according to additional negative prognostic factors. Our objective was to evaluate the biomarker potential of the ASRGL1 protein in endometrial carcinoma. Using The Human Protein Atlas (www.proteinatlas.org), the l-asparaginase (ASRGL1) protein was identified as an endometrial carcinoma biomarker candidate. ASRGL1 expression was immunohistochemically evaluated with an extensively validated antibody on two independent endometrial carcinoma cohorts (n=229 and n=286) arranged as tissue microarrays. Staining results were correlated with clinical features. Reduced expression of ASRGL1, defined as <75% positively stained tumor cells, was significantly associated with poor prognosis and reduced disease-specific survival in endometrioid endometrial adenocarcinoma (EEA). In multivariate analysis the hazard ratios for disease-specific survival were 3.55 (95% CI=1.10-11.43; p=0.003) and 3.23 (95% CI=1.53-6.81; p=0.002) in the two cohorts, respectively. Of the 48 cases with Grade 3 Stage I tumor all disease-related deaths were associated with low ASRGL1 expression. Loss of ASRGL1 in EEA is a powerful biomarker for poor prognosis and retained ASRGL1 has a positive impact on survival. ASRGL1 immunohistochemistry has potential to become an additional tool for prognostication in cases where tailoring adjuvant treatment according to additional prognostic factors besides grade and stage is recommended.


Spherical particle size determination by analytical inversion of the UV-visible-NIR extinction spectrum.
An analytic inversion method, based on the anomalous diffraction approximation for nonabsorbing spherical particles, was developed to retrieve the size distribution from the optical turbidity or extinction spectrum. This method makes use of a differential Fourier cosine transform approach and provides a simple and fast inversion by means of fast Fourier transform and the Savitzky-Golay filter. The applicability of this algorithm was tested on the extinction data generated by the Mie solution. The effects of noise, modality, band limits, and data set size were analyzed by comparison with simulated data. This method can be used to reconstruct the original monomodal and bimodal distributions from 10% noise-corrupted data. The peak position and ratio of peak heights can be recovered with 10% or less deviation. The experiments with latex spheres showed that the inversion result from this method compares favorably with that from the dynamic light scattering measurement.


Preexisting domain motions underlie protonation-dependent structural transitions of the P-type Ca2+-ATPase.
We have performed microsecond molecular dynamics (MD) simulations to determine the mechanism for protonation-dependent structural transitions of the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA), one of the most prominent members of the large P-type ATPase superfamily that transports ions across biological membranes. The release of two H+ from the transport sites activates SERCA by inducing a structural transition between low (E2) and high (E1) Ca2+-affinity states (E2-to-E1 transition), but the structural mechanism by which transport site deprotonation facilitates this transition is unknown. We performed microsecond all-atom MD simulations to determine the effects of transport site protonation on the structural dynamics of the E2 state in solution. We found that the protonated E2 state has structural characteristics that are similar to those observed in crystal structures of E2. Upon deprotonation, a single Na+ ion rapidly (<10 ns) binds to the transmembrane transport sites and induces a kink in M5, disrupts the M3-M5 interface, and increases the mobility of the M3/A-M3 linker. Principal component analysis showed that counter-rotation of the cytosolic N-A domains about the membrane normal axis, which is the primary motion driving the E2-to-E1 transition, is present in both protonated and deprotonated E2 states; however, protonation-dependent structural changes in the transmembrane domain control the hierarchical organization and amplitude of this motion. We propose that preexisting rigid-body domain motions underlie structural transitions of SERCA, where the functionally important directionality is preserved while transport site protonation controls the dominance and amplitude of motion to shift the equilibrium between the E1 and E2 states. We conclude that ligand-induced modulation of preexisting domain motions is likely a common theme in structural transitions of the P-type ATPase superfamily.


Readmission after colectomy for cancer predicts one-year mortality.
Early hospital readmission is a common and costly problem in the Medicare population. In 2009, the Centers for Medicaid and Medicare Services began mandating hospital reporting of disease-specific readmission rates. We sought to determine the rate and predictors of readmission after colectomy for cancer, as well as the association between readmission and mortality. Medicare beneficiaries who underwent colectomy for stage I to III colon adenocarcinoma from 1992 to 2002 were identified from the Surveillance, Epidemiology, and End Results-Medicare database. Multivariate logistic regression identified predictors of early readmission and 1-year mortality. Odds ratios were adjusted for multiple factors, including measures of comorbidity, socioeconomic status, and disease severity. Of 42,348 patients who were discharged, 4662 (11.0%) were readmitted within 30 days. The most common causes of rehospitalization were ileus/obstruction and infection. Significant predictors of readmission included male gender, comorbidity, emergent admission, prolonged hospital stay, blood transfusion, ostomy, and discharge to nursing home. Readmission was inversely associated with hospital procedure volume, but not surgeon volume. After adjusting for potential confounding variables, the predicted probability of 1-year mortality was 16% for readmitted patients, compared with 7% for those not readmitted. This difference in mortality was significant for all stages of cancer. Early readmission after colectomy for cancer is common and due in part to modifiable factors. There is a remarkable association between readmission and 1-year mortality. Early readmission is therefore an important quality-of-care indicator for colon cancer surgery. These findings may facilitate the development of targeted interventions that will decrease readmissions and improve patient outcomes.


Continuous flow Overhauser dynamic nuclear polarization of water in the fringe field of a clinical magnetic resonance imaging system for authentic image contrast.
We describe and demonstrate a system to generate hyperpolarized water in the 0.35 T fringe field of a clinical 1.5 T whole-body magnetic resonance imaging (MRI) magnet. Once generated, the hyperpolarized water is quickly and continuously transferred from the 0.35 T fringe to the 1.5 T center field of the same magnet for image acquisition using standard MRI equipment. The hyperpolarization is based on Overhauser dynamic nuclear polarization (DNP), which effectively and quickly transfers the higher spin polarization of free radicals to nuclear spins at ambient temperatures. We visualize the dispersion of hyperpolarized water as it flows through water-saturated systems by utilizing an observed -15-fold DNP signal enhancement with respect to the unenhanced (1)H MRI signal of water at 1.5 T. The experimental DNP apparatus presented here is readily portable and can be brought to and used with any conventional unshielded MRI system. A new method of immobilizing radicals to gel beads via polyelectrolyte linker arms is described, which led to superior flow Overhauser DNP performance compared to previously presented gels. We discuss the general applicability of Overhauser DNP of water and aqueous solutions in the fringe field of commercially available magnets with central fields up to 4.7 T.


Effects of a minimal intervention to increase physical activity in women: daily activity records.
Effective interventions to increase physical activity levels are critical in a nation where inactivity is a national public health problem. This pilot study examined whether a minimal intervention (daily records of physical activity) increased activity levels in a community sample of working women. In a longitudinal, pretest-posttest design, 49 working women were randomly assigned at the work site level to the control (n = 25) or intervention group (n = 24). At pretest and posttest, subjects completed self-report questionnaires that measured psychological, social-environmental, physical activity, and demographic variables. Subjects in the intervention group kept daily records of their physical activities during the 12-week study, while those in the control group kept no records. In order to compare activity in the two groups, all subjects wore pedometers daily that recorded number of steps. There was a significant difference between groups in the pedometer values (mean number of daily steps) at the end of the study period (mean difference +/- SE 2147 +/- 636, p = .022) (2000 steps = approximately 1 mile). Multiple regression analysis showed that only the intervention (p = .003) was a significant predictor of the pedometer values. Hierarchical data analysis was used to account for the intra-class correlation of 0.48 within work site. Results from this sample of 49 women indicated that mean activity was greater in the intervention group compared to the control group. Recording daily activity is a cost-effective and acceptable intervention that may increase activity levels in women. However, more research is recommended to study the dual role of activity records as a data collection method as well as a potential intervention to increase physical activity.


Understanding the relative importance of preserving functional abilities in Alzheimer's disease in the United States and Germany.
To estimate the relative importance that Alzheimer's disease (AD) caregivers in the United States and Germany place on preserving patients' ability to perform activities of daily living. US and German residents providing care for a person with AD completed an online preference survey. Each respondent completed five best-worst scaling questions. Each question related to five of 10 activities from the Disability Assessment for Dementia scale. Preference weights, indicating the relative importance of preserving the ability to perform these 10 activities for 36 months, were estimated using maximum-difference scaling. A separate model was estimated for each country. Four hundred and three US and 400 German caregivers completed the survey. In both countries, preserving a patients' ability to use the toilet without accidents was the most important activity and handling money was the least important activity. There were few differences between US and German caregivers in the relative importance across activities. Caregivers generally placed greater importance on preserving basic activities of daily living than on preserving instrumental activities of daily living. Understanding differences in the relative importance of functional items in the DAD may contribute to a better understanding of the benefits of different AD treatment and support measures.


Toward an interactive article: integrating journals and biological databases.
Journal articles and databases are two major modes of communication in the biological sciences, and thus integrating these critical resources is of urgent importance to increase the pace of discovery. Projects focused on bridging the gap between journals and databases have been on the rise over the last five years and have resulted in the development of automated tools that can recognize entities within a document and link those entities to a relevant database. Unfortunately, automated tools cannot resolve ambiguities that arise from one term being used to signify entities that are quite distinct from one another. Instead, resolving these ambiguities requires some manual oversight. Finding the right balance between the speed and portability of automation and the accuracy and flexibility of manual effort is a crucial goal to making text markup a successful venture. We have established a journal article mark-up pipeline that links GENETICS journal articles and the model organism database (MOD) WormBase. This pipeline uses a lexicon built with entities from the database as a first step. The entity markup pipeline results in links from over nine classes of objects including genes, proteins, alleles, phenotypes and anatomical terms. New entities and ambiguities are discovered and resolved by a database curator through a manual quality control (QC) step, along with help from authors via a web form that is provided to them by the journal. New entities discovered through this pipeline are immediately sent to an appropriate curator at the database. Ambiguous entities that do not automatically resolve to one link are resolved by hand ensuring an accurate link. This pipeline has been extended to other databases, namely Saccharomyces Genome Database (SGD) and FlyBase, and has been implemented in marking up a paper with links to multiple databases. Our semi-automated pipeline hyperlinks articles published in GENETICS to model organism databases such as WormBase. Our pipeline results in interactive articles that are data rich with high accuracy. The use of a manual quality control step sets this pipeline apart from other hyperlinking tools and results in benefits to authors, journals, readers and databases.


Nanoporous polymers for hydrogen storage.
The design of hydrogen storage materials is one of the principal challenges that must be met before the development of a hydrogen economy. While hydrogen has a large specific energy, its volumetric energy density is so low as to require development of materials that can store and release it when needed. While much of the research on hydrogen storage focuses on metal hydrides, these materials are currently limited by slow kinetics and energy inefficiency. Nanostructured materials with high surface areas are actively being developed as another option. These materials avoid some of the kinetic and thermodynamic drawbacks of metal hydrides and other reactive methods of storing hydrogen. In this work, progress towards hydrogen storage with nanoporous materials in general and porous organic polymers in particular is critically reviewed. Mechanisms of formation for crosslinked polymers, hypercrosslinked polymers, polymers of intrinsic microporosity, and covalent organic frameworks are discussed. Strategies for controlling hydrogen storage capacity and adsorption enthalpy via manipulation of surface area, pore size, and pore volume are discussed in detail.


Inflammatory bowel disease and prostate cancer risk: A systematic review.
Objective: To evaluate the risk of prostate cancer (PCa) in patients with inflammatory bowel disease (IBD), focussing on ulcerative colitis (UC) and Crohn's disease (CD) separately. Methods: A systemic search was carried out using PubMed and Web of Science databases following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We retrieved a total of 349 articles. All the articles were in the English language and investigated the incidence of PCa in patients with IBD. Results: Nine studies met our inclusion criteria, with a total of 205 037 men. Two studies reported an increase in the risk of PCa in men with IBD in general. Five other studies reported an increased risk of PCa in men with UC or with CD specifically. On the other hand, two studies reported a decreased risk of PCa in patients with UC and patients with IBD treated with aminosalicylates. Conclusions: While men with UC appear to have higher risk of developing PCa, data on patients with CD are inconclusive. Therefore, patients with UC may benefit from earlier PCa screening. Our findings confirm a complex interplay between IBD and PCa, including factors such as genetic predisposition, systemic inflammation and treatment effects. The modulatory effect of treatment strategies for IBD on the development and progression of PCa might be of clinical significance. Abbreviations: CD: Crohn's disease; CRP: C- reactive protein; FOLH1: folate hydrolase 1; GIT: gastrointestinal tract; IBD: inflammatory bowel disease; IL-6: interleukin 6; NOS: Newcastle-Ottawa Scale; PCa: prostate cancer; PRISMA: Preferred Reporting Items for Systematic Reviews and Meta-Analyses; PSMA: prostate-specific membrane antigen; UC: ulcerative colitis.


Glucagon receptor antagonist and GIP agonist combination for diet-induced obese mice.
Ablation of glucagon receptor signaling represents a potential treatment option for type 2 diabetes (T2DM). Additionally, activation of glucose-dependent insulinotropic polypeptide (GIP) receptor signaling also holds therapeutic promise for T2DM. Therefore, this study examined both independent and combined metabolic actions of desHis(1)Pro(4)Glu(9)(Lys(12)PAL)-glucagon (glucagon receptor antagonist) and d-Ala(2)GIP (GIP receptor agonist) in diet-induced obese mice. Glucagon receptor binding has been linked to alpha-helical structure and desHis(1)Pro(4)Glu(9)(Lys(12)PAL)-glucagon displayed enhanced alpha-helical content compared with native glucagon. In clonal pancreatic BRIN-BD11 beta-cells, desHis(1)Pro(4)Glu(9)(Lys(12)PAL)-glucagon was devoid of any insulinotropic or cAMP-generating actions, and did not impede d-Ala(2)GIP-mediated (P<0.01 to P<0.001) effects on insulin and cAMP production. Twice-daily injection of desHis(1)Pro(4)Glu(9)(Lys(12)PAL)-glucagon or d-Ala(2)GIP alone, and in combination, in high-fat-fed mice failed to affect body weight or energy intake. Circulating blood glucose levels were significantly (P<0.05 to P<0.01) decreased by all treatments regimens, with plasma and pancreatic insulin elevated (P<0.05 to P<0.001) in all mice receiving d-Ala(2)GIP. Interestingly, plasma glucagon concentrations were decreased (P<0.05) by sustained glucagon inhibition (day 28), but increased (P<0.05) by d-Ala(2)GIP therapy, with a combined treatment resulting in glucagon concentration similar to saline controls. All treatments improved (P<0.01) intraperitoneal and oral glucose tolerance, and peripheral insulin sensitivity. d-Ala(2)GIP-treated mice showed increased glucose-induced insulin secretion in response to intraperitoneal and oral glucose. Metabolic rate and ambulatory locomotor activity were increased (P<0.05 to P<0.001) in all desHis(1)Pro(4)Glu(9)(Lys(12)PAL)-glucagon-treated mice. These studies highlight the potential of glucagon receptor inhibition alone, and in combination with GIP receptor activation, for T2DM treatment.


Vitamin C in Stem Cell Biology: Impact on Extracellular Matrix Homeostasis and Epigenetics.
Transcription factors and signaling molecules are well-known regulators of stem cell identity and behavior; however, increasing evidence indicates that environmental cues contribute to this complex network of stimuli, acting as crucial determinants of stem cell fate. l-Ascorbic acid (vitamin C (VitC)) has gained growing interest for its multiple functions and mechanisms of action, contributing to the homeostasis of normal tissues and organs as well as to tissue regeneration. Here, we review the main functions of VitC and its effects on stem cells, focusing on its activity as cofactor of Fe+2/αKG dioxygenases, which regulate the epigenetic signatures, the redox status, and the extracellular matrix (ECM) composition, depending on the enzymes' subcellular localization. Acting as cofactor of collagen prolyl hydroxylases in the endoplasmic reticulum, VitC regulates ECM/collagen homeostasis and plays a key role in the differentiation of mesenchymal stem cells towards osteoblasts, chondrocytes, and tendons. In the nucleus, VitC enhances the activity of DNA and histone demethylases, improving somatic cell reprogramming and pushing embryonic stem cell towards the naive pluripotent state. The broad spectrum of actions of VitC highlights its relevance for stem cell biology in both physiology and disease.


Serum Dickkopf-1 as a Biomarker for the Diagnosis of Hepatocellular Carcinoma.
Dickkopf-1 (DKK-1) is a Wnt/β-catenin signaling pathway inhibitor. We investigated whether DKK-1 is related to progression in hepatocellular carcinoma (HCC) cells and HCC patients. In vitro reverse-transcription polymerase chain reaction (RT-PCR), wound healing assays, invasion assays, and ELISAs of patient serum samples were employed. The diagnostic accuracy of the serum DKK-1 ELISA was assessed using receiver operating characteristic (ROC) curves and area under ROC (AUC) analyses. RT-PCR showed high DKK-1 expression in Hep3B and low in 293 cells. Similarly, the secreted DKK-1 concentration in the culture media was high in Hep3B and low in 293 cells. Wound healing and invasion assays using 293, Huh7, and Hep3B cells showed that DKK-1 overexpression promoted cell migration and invasion, whereas DKK-1 knock-down inhibited them. When serum DKK-1 levels were assessed in 370 participants (217 with HCC and 153 without), it was significantly higher in HCC patients than in control groups (median 1.48 ng/mL vs. 0.90 ng/mL, p<0.001). The optimum DKK-1 cutoff level was 1.01 ng/mL (AUC=0.829; sensitivity 90.7%; specificity 62.0%). Although DKK-1 had a higher AUC than alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) (AUC=0.829 vs. 0.794 and 0.815, respectively), they were statistically similar (all p>0.05). When three biomarkers were combined (DKK-1 plus AFP plus DCP), they showed significantly higher AUC (AUC=0.952) than single marker, DKK-1 plus AFP, or DKK-1 plus DCP (all p<0.001). DKK-1 might be a key regulator in HCC progression and a potential therapeutic target in HCC. Serum DKK-1 could complement the diagnostic accuracy of AFP and DCP.


Application of continual injection liquid-phase microextraction method coupled with liquid chromatography to the analysis of organophosphorus pesticides.
A liquid-phase microextraction coupled with LC method has been developed for the determination of organophosphorus pesticides (methidation, quinalphos and profenofos) in drinking water samples. In this method, a small amount (3 microL) of isooctane as the acceptor phase was introduced continually to fill-up the channel of a 1.5 cm polypropylene hollow fiber using a microsyringe while the hollow fiber was immersed in an aqueous donor solution. A portion of the acceptor phase (ca. 0.4 microL) was first introduced into the hollow fiber and additional amounts (ca. 0.2 microL) of the acceptor phase were introduced to replenish at intervals of 3 min until set end of extraction (40 min). After extraction, the acceptor phase was withdrawn and transferred into a 2 mL vial for a drying step prior to injection into a LC system. Parameters that affect the extraction efficiency were studied including the organic solvent, length of fiber, volume of acceptor and donor phase, stirring rate, extraction time, and effect of salting out. The proposed method provided good enrichment factors of up to 189.50, with RSD ranging from 0.10 to 0.29%, analyte recoveries of over 79.80% and good linearity ranging from 10.0 to 1.25 mg/L. The LOD ranged from 2.86 to 82.66 microg/L. This method was applied successfully to the determination of organophosphorus pesticides in selected drinking water samples.


Genome-wide association study identifies eight loci associated with blood pressure.
Elevated blood pressure is a common, heritable cause of cardiovascular disease worldwide. To date, identification of common genetic variants influencing blood pressure has proven challenging. We tested 2.5 million genotyped and imputed SNPs for association with systolic and diastolic blood pressure in 34,433 subjects of European ancestry from the Global BPgen consortium and followed up findings with direct genotyping (N ≤ 71,225 European ancestry, N ≤ 12,889 Indian Asian ancestry) and in silico comparison (CHARGE consortium, N = 29,136). We identified association between systolic or diastolic blood pressure and common variants in eight regions near the CYP17A1 (P = 7 × 10(-24)), CYP1A2 (P = 1 × 10(-23)), FGF5 (P = 1 × 10(-21)), SH2B3 (P = 3 × 10(-18)), MTHFR (P = 2 × 10(-13)), c10orf107 (P = 1 × 10(-9)), ZNF652 (P = 5 × 10(-9)) and PLCD3 (P = 1 × 10(-8)) genes. All variants associated with continuous blood pressure were associated with dichotomous hypertension. These associations between common variants and blood pressure and hypertension offer mechanistic insights into the regulation of blood pressure and may point to novel targets for interventions to prevent cardiovascular disease.


Ondansetron for acute gastroenteritis in children.
In my pediatric practice I see many children with acute gastroenteritis. Their parents ask for antiemetic medications. Ondansetron has been well tolerated when used to control nausea and vomiting in patients receiving chemotherapy. Is there a role for it in managing acute gastroenteritis in children? Use of antiemetics is not indicated for treatment of acute gastroenteritis. Some evidence suggests ondansetron is clinically more effective and better tolerated and has a better side effect profile than other antiemetics, but does not suggest that it reduces hospital admission rates. Use of ondansetron, as with other antiemetics, continues to be at treating physicians' discretion, and potential adverse events should be considered before administration.


Proton NMR spectra of urine as indicators of renal damage. Mercury-induced nephrotoxicity in rats.
Rats were injected intraperitoneally with HgCl2 at doses of 2.5, 5, 7.5, and 10 mumol of Hg/kg. Urine was collected over a 24-hr period. At this time, plasma samples were taken and kidney damage was assessed by histological examination. Urinary gamma-glutamyltransferase levels were significantly elevated at Hg2+ doses of 7.5 and 10 mumol/kg, consistent with the detection of acute tubular necrosis by light microscopy. Resonances for a large number of low molecular weight metabolites were assigned in high resolution 1H NMR spectra of rat urine. Spectra from small volumes of urine (about 0.5 ml) were obtained in less than 5 min with no pretreatment. Significant Hg2+ dose-related decreases in the excretion of creatinine and citrate and increases of glucose, glycine, alanine, alpha-ketoglutarate, succinate, and acetate were detected. Elevated levels of lactate and creatinine in plasma of rats receiving the two highest doses were found by 1H NMR. There was a good correspondence between the histopathology, enzyme excretion, and 1H NMR urinary metabolite fingerprints in the assessment of Hg2+-induced renal damage. 1H NMR provided a sensitive measure of mercury-induced nephrotoxic lesions, and information on the molecular basis of mercury cytotoxicity was derived from the abnormal patterns of metabolite excretion. These suggested that primary metabolic effects of mercury were upon mitochondrial metabolism, in particular inhibition of certain citric acid cycle enzymes leading to decreased utilization of alpha-ketoglutarate and succinate by the renal tubular cells. The decrease in urinary citrate associated with Hg2+ dosing was attributed to intracellular, tubular acidosis with concomitant enhanced citrate reabsorption. The acidosis was assumed to arise from a combination of the inhibition of tubular carbonic anhydrase and a mild metabolic lactic acidosis due to increased activity of anaerobic pathways in the kidney. The possible extension of the 1H NMR techniques to the investigation of the nephrotoxic potential of other compounds and drugs is discussed.


Community perspectives on the benefits and risks of technologically enhanced communicable disease surveillance systems: a report on four community juries.
Outbreaks of infectious disease cause serious and costly health and social problems. Two new technologies - pathogen whole genome sequencing (WGS) and Big Data analytics - promise to improve our capacity to detect and control outbreaks earlier, saving lives and resources. However, routinely using these technologies to capture more detailed and specific personal information could be perceived as intrusive and a threat to privacy. Four community juries were convened in two demographically different Sydney municipalities and two regional cities in New South Wales, Australia (western Sydney, Wollongong, Tamworth, eastern Sydney) to elicit the views of well-informed community members on the acceptability and legitimacy of: making pathogen WGS and linked administrative data available for public health researchusing this information in concert with data linkage and machine learning to enhance communicable disease surveillance systems Fifty participants of diverse backgrounds, mixed genders and ages were recruited by random-digit-dialling and topic-blinded social-media advertising. Each jury was presented with balanced factual evidence supporting different expert perspectives on the potential benefits and costs of technologically enhanced public health research and communicable disease surveillance and given the opportunity to question experts. Almost all jurors supported data linkage and WGS on routinely collected patient isolates for the purposes of public health research, provided standard de-identification practices were applied. However, allowing this information to be operationalised as a syndromic surveillance system was highly contentious with three juries voting in favour, and one against by narrow margins. For those in favour, support depended on several conditions related to system oversight and security being met. Those against were concerned about loss of privacy and did not trust Australian governments to run secure and effective systems. Participants across all four events strongly supported the introduction of data linkage and pathogenomics to public health research under current research governance structures. Combining pathogen WGS with event-based data surveillance systems, however, is likely to be controversial because of a lack of public trust, even when the potential public health benefits are clear. Any suggestion of private sector involvement or commercialisation of WGS or surveillance data was unanimously rejected.


Neural responses to incongruency in a blocked-trial Stroop fMRI task in major depressive disorder.
Patients with major depressive disorder (MDD) perform poorly on the Stroop task, which is a measure of the executive control of attention, with impaired interference resolution. The neural correlates of this deficit are not well described. To examine how this deficit relates to pathophysiological abnormalities in MDD, we conducted an fMRI Stroop study comparing MDD subjects to controls. Forty-two unmedicated patients with current MDD and 17 control subjects underwent fMRI scanning with a color-word Stroop task. Subjects assessed font color during alternating color identification (e.g., 'XXXX' in blue) and incongruent color/word blocks (e.g., the word 'red' in blue). We examined neural activation that was greater in incongruent than color identification blocks (Z>2.3 and corrected p<0.05), controlling for trial-by-trial reaction time. Compared to controls, MDD subjects exhibited lower activation during incongruent blocks across multiple brain regions, including middle frontal gyrus, paracingulate and posterior cingulate, precuneus, occipital regions, and brain stem. No brain regions were identified in which MDD subjects were more active than controls during incongruent blocks. Not all MDD subjects were antidepressant-naïve. Brain regions related to executive function, visual processing, and semantic processing are less active during processing of incongruent stimuli in MDD subjects as compared to controls. Deficits of attention in MDD may be the product of a failure to maintain activity across a distributed network in a sustained manner, as is required over the sequential trials in this block design. Further studies may clarify whether the abnormalities represent a trait or state deficit.


Emergency hospital admissions associated with a non-randomised housing intervention meeting national housing quality standards: a longitudinal data linkage study.
We investigated tenant healthcare utilisation associated with upgrading 8558 council houses to a national quality standard. Homes received multiple internal and external improvements and were analysed using repeated measures of healthcare utilisation. The primary outcome was emergency hospital admissions for cardiorespiratory conditions and injuries for residents aged 60 years and over. Secondary outcomes included each of the separate conditions, for tenants aged 60 and over, and for all ages. Council home address and intervention records for eight housing cointerventions were anonymously linked to demographic data, hospital admissions and deaths for individuals in a dynamic cohort. Counts of health events were analysed using multilevel regression models to investigate associations between receipt of each housing improvement, adjusting for potential confounding factors and regional trends. Residents aged 60 years and over living in homes when improvements were made were associated with up to 39% fewer admissions compared with those living in homes that were not upgraded (incidence rate ratio=0.61, 95% CI 0.53 to 0.72). Reduced admissions were associated with electrical systems, windows and doors, wall insulation, and garden paths. There were small non-significant reductions for the primary outcome associated with upgrading heating, adequate loft insulation, new kitchens and new bathrooms. Results suggest that hospital admissions can be avoided through improving whole home quality standards. This is the first large-scale longitudinal evaluation of a whole home intervention that has evaluated multiple improvement elements using individual-level objective routine health data.


In vivo testing of a novel adjustable glaucoma drainage device.
We report on the in vivo testing of a novel noninvasively adjustable glaucoma drainage device (AGDD), which features an adjustable outflow resistance, and assess the safety and efficiency of this implant. Under general anesthesia, the AGDD was implanted on seven white New Zealand rabbits for a duration of 4 months under a scleral flap in a way analogous to the Ex-PRESS device and set in an operationally closed position. The IOP was measured on a regular basis on the operated and control eyes using a rebound tonometer. Once a month the AGDD was adjusted noninvasively from its fully closed to its fully open position and the resulting pressure drop was measured. The contralateral eye was not operated and served as control. After euthanization, the eyes were collected for histology evaluation. The mean preoperative IOP was 11.1 ± 2.4 mm Hg. The IOP was significantly lower for the operated eye (6.8 ± 2 mm Hg) compared to the nonoperated eye (13.1 ± 1.6 mm Hg) during the first 8 days after surgery. When opening the AGDD from its fully closed to fully open position, the IOP dropped significantly from 11.2 ± 2.9 to 4.8 ± 0.9 mm Hg (P < 0.05). Implanting the AGDD is a safe and uncomplicated surgical procedure. The fluidic resistance was noninvasively adjustable during the postoperative period with the AGDD between its fully closed and fully open positions.


Comparison of the peptidase activity in the oncosphere excretory/secretory products of Taenia solium and Taenia saginata.
We compared the peptidase activities of the excretory/secretory (E/S) antigens of oncospheres of Taenia solium and related, but nonpathogenic, Taenia saginata. Taenia solium and T. saginata oncospheres were cultured, and the spent media of 24-, 48-, 72-, and 96-hr fractions were analyzed. Activities for serine peptidases (chymotrypsin-, trypsin-, and elastase-like), cysteine peptidases (cathepsin B-, cathepsin L-, and calpaine-like), and aminopeptidase (B-like peptidases) were tested fluorometrically with peptides coupled to 7-amino-4-methylcoumarin. In both species, the E/S antigens showed cysteine, serine, and aminopeptidase activities. Although no particular peptidase had high activity in T. solium, and was absent in T. saginata, or vice versa, different patterns of activity were found. A chymotrypsin-like peptidase showed the highest activity in both parasites, and it had 10 times higher activity in T. solium than in T. saginata. Trypsin-like and cathepsin B-like activities were significantly higher in T. solium. Minimal levels of cathepsin B were present in both species, and higher levels of elastase-like and cathepsin L-like activity were observed in T. saginata. Taenia solium and T. saginata have different levels and temporal activities of proteolytic enzymes that could play a modulator role in the host specificity for larval invasion through penetration of the intestinal mucosa.


Climate change reduces resilience to fire in subalpine rainforests.
Climate change is affecting the distribution of species and the functioning of ecosystems. For species that are slow growing and poorly dispersed, climate change can force a lag between the distributions of species and the geographic distributions of their climatic envelopes, exposing species to the risk of extinction. Climate also governs the resilience of species and ecosystems to disturbance, such as wildfire. Here we use species distribution modelling and palaeoecology to assess and test the impact of vegetation-climate disequilibrium on the resilience of an endangered fire-sensitive rainforest community to fires. First, we modelled the probability of occurrence of Athrotaxis spp. and Nothofagus gunnii rainforest in Tasmania (hereon "montane rainforest") as a function of climate. We then analysed three pollen and charcoal records spanning the last 7,500 cal year BP from within both high (n = 1) and low (n = 2) probability of occurrence areas. Our study indicates that climatic change between 3,000 and 4,000 cal year bp induced a disequilibrium between montane rainforests and climate that drove a loss of resilience of these communities. Current and future climate change are likely to shift the geographic distribution of the climatic envelopes of this plant community further, suggesting that current high-resilience locations will face a reduction in resilience. Coupled with the forecast of increasing fire activity in southern temperate regions, this heralds a significant threat to this and other slow growing, poorly dispersed and fire sensitive forest systems that are common in the southern mid to high latitudes.


Lipopeptides: from self-assembly to bioactivity.
This Feature Article discusses several classes of lipopeptide with important biomedical applications as antimicrobial and antifungal agents, in immune therapies and in personal care applications among others. Two main classes of lipopeptide are considered: (i) bacterially-expressed lipopeptides with a cyclic peptide headgroup and (ii) linear lipopeptides (with one or more lipid chains) based on bio-derived and bio-inspired amino acid sequences with current clinical applications. The applications are briefly summarized, and the biophysical characterization of the molecules is reviewed, with a particular focus on self-assembly. For several of these types of biomolecule, the formation of micelles above a critical micelle concentration has been observed while others form bilayer structures, depending on conditions of pH and temperature. As yet, there are few studies on the possible relationship between self-assembly into structures such as micelles and bioactivity of this class of molecule although this is likely to attract further attention.


Sequential interplay of nicotinic and GABAergic signaling guides neuronal development.
GABA (gamma-aminobutyric acid), the major inhibitory transmitter in the brain, goes through a transitory phase of excitation during development. The excitatory phase promotes neuronal growth and integration into circuits. We show here that spontaneous nicotinic cholinergic activity is responsible for terminating GABAergic excitation and initiating inhibition. It does so by changing chloride transporter levels, shifting the driving force on GABA-induced currents. The timing of the transition is critical, because the two phases of GABAergic signaling provide contrasting developmental instructions. Synergistic with nicotinic excitation, GABAergic inhibition constrains neuronal morphology and innervation. The results reveal a multitiered activity-dependent strategy controlling neuronal development.


Sleep-wake classification via quantifying heart rate variability by convolutional neural network.
Fluctuations in heart rate are intimately related to changes in the physiological state of the organism. We exploit this relationship by classifying a human participant's wake/sleep status using his instantaneous heart rate (IHR) series. We use a convolutional neural network (CNN) to build features from the IHR series extracted from a whole-night electrocardiogram (ECG) and predict every 30 s whether the participant is awake or asleep. Our training database consists of 56 normal participants, and we consider three different databases for validation; one is private, and two are public with different races and apnea severities. On our private database of 27 participants, our accuracy, sensitivity, specificity, and [Formula: see text] values for predicting the wake stage are [Formula: see text], 52.4%, 89.4%, and 0.83, respectively. Validation performance is similar on our two public databases. When we use the photoplethysmography instead of the ECG to obtain the IHR series, the performance is also comparable. A robustness check is carried out to confirm the obtained performance statistics. This result advocates for an effective and scalable method for recognizing changes in physiological state using non-invasive heart rate monitoring. The CNN model adaptively quantifies IHR fluctuation as well as its location in time and is suitable for differentiating between the wake and sleep stages.


Homer1/mGluR5 activity moderates vulnerability to chronic social stress.
Stress-induced psychiatric disorders, such as depression, have recently been linked to changes in glutamate transmission in the central nervous system. Glutamate signaling is mediated by a range of receptors, including metabotropic glutamate receptors (mGluRs). In particular, mGluR subtype 5 (mGluR5) is highly implicated in stress-induced psychopathology. The major scaffold protein Homer1 critically interacts with mGluR5 and has also been linked to several psychopathologies. Yet, the specific role of Homer1 in this context remains poorly understood. We used chronic social defeat stress as an established animal model of depression and investigated changes in transcription of Homer1a and Homer1b/c isoforms and functional coupling of Homer1 to mGluR5. Next, we investigated the consequences of Homer1 deletion, overexpression of Homer1a, and chronic administration of the mGluR5 inverse agonist CTEP (2-chloro-4-((2,5-dimethyl-1-(4-(trifluoromethoxy)phenyl)-1H-imidazol-4-yl)ethynyl)pyridine) on the effects of chronic stress. In mice exposed to chronic stress, Homer1b/c, but not Homer1a, mRNA was upregulated and, accordingly, Homer1/mGluR5 coupling was disrupted. We found a marked hyperactivity behavior as well as a dysregulated hypothalamic-pituitary-adrenal axis activity in chronically stressed Homer1 knockout (KO) mice. Chronic administration of the selective and orally bioavailable mGluR5 inverse agonist, CTEP, was able to recover behavioral alterations induced by chronic stress, whereas overexpression of Homer1a in the hippocampus led to an increased vulnerability to chronic stress, reflected in an increased physiological response to stress as well as enhanced depression-like behavior. Overall, our results implicate the glutamatergic system in the emergence of stress-induced psychiatric disorders, and support the Homer1/mGluR5 complex as a target for the development of novel antidepressant agents.


Epitope-relatedness and phenotyping of hepatic cytochromes P-450 with monoclonal antibodies.
The epitope-specific cytochrome P-450 content of animal livers was analysed by radioimmunoassay using a panel of seven monoclonal antibodies (MAbs) made to a 3-methylcholanthrene-induced rat liver cytochrome P-450. Competitive radioimmunoassays utilizing a reference radiolabelled MAb and a series of unlabelled MAbs indicated that there are at least three distinct classes of MAbs to different epitopes on cytochrome P-450. In addition, a direct radioimmunoassay employing a radiolabelled second antibody detected MAb-specific cytochromes P-450 in livers from different animals. This radioimmunoassay detected large elevations in the levels of these cytochromes P-450 in the livers of 3-methylcholanthrene-treated rats and C57BL/6 mice compared with untreated rats, 3-methylcholanthrene-treated DBA/2 mice or guinea pigs. The two complementary radioimmunoassay methods are sensitive, efficient, and easily applicable for screening large number of tissue samples for MAb-defined cytochrome P-450 phenotype.


Genetics of emergent suicidality during antidepressive treatment--data from a naturalistic study on a large sample of inpatients with a major depressive episode.
Factors contributing to treatment-emergent suicidal ideation (TESI) using antidepressants have been in the focus of recent research strategies. We investigated previously established clinical predictors of TESI and combined these with several polymorphisms of candidate genes in patients with major depressive disorder. Common polymorphisms involved in the tryptophan hydroxylase 1 (TPH1) and 2 (TPH2), serotonin transporter, monoamine oxidase A (MAOA) and brain-derived neurotrophic factor (BDNF) were investigated in a naturalistic inpatient study of the German research network on depression. We compared patients showing TESI with non-TESI suicidal patients and with non-suicidal patients using univariate tests to detect relevant factors, which were further tested in logistic regression and CART (Classification and Regression Trees) analyses. Of the 269 patients, TESI occurred in 22 patients (17 female), 117 patients were defined as non-TESI suicidal patients, and 130 patients were classified as non-suicidal. When comparing cases with both control groups we found the TPH2 rs1386494 (C/T) polymorphism to be moderately associated with TESI (Univariate tests: TESI vs. non-suicidality: p=0.005; adjusted: p=0.09; TESI vs. non-TESI suicidal patients: p=0.0024; adjusted: p=0.086). This polymorphism remained the only significant genetic factor in addition to clinical predictors in logistic regression and CART analyses. CART analyses suggested interactions with several clinical predictors. Haplotype analyses further supported a contribution of this polymorphism in TESI. The TPH2 rs1386494 (C/T) polymorphism might contribute to the genetic background of TESI. This polymorphism has been previously associated with committed suicide and major depressive disorder. The small number of cases warrants replication in larger patient samples. Lack of a placebo control group hampers definite conclusions on an association with antidepressive treatment.


A systematic review of the evidence on clitoral reconstruction after female genital mutilation/cutting.
Clitoral reconstruction is a new surgical technique for women who have undergone female genital mutilation/cutting (FGM/C). To review evidence on the safety and efficacy of clitoral reconstruction. PubMed and Cochrane databases were searched for articles published in any language from database inception until May 2014. Search terms related to FGM/C and clitoral reconstruction were used in various combinations. Studies of any design that reported on safety or clinical outcomes (e.g. appearance, pain, sexual response, or patient satisfaction) associated with clitoral reconstruction after FGM/C were included. Evidence was summarized and systematically assessed via a standard data abstraction form. Four of 269 identified articles were included. They were fair to poor in quality. Summary measures could not be computed owing to heterogeneity. The studies reported on immediate surgical complications, clitoral appearance, dyspareunia or chronic pain, and clitoral function postoperatively via non-standardized scales. Women who request clitoral reconstruction should be informed about the scarcity of evidence available. Additional research is needed on the safety and efficacy of the procedure to identify both long-term outcomes and which women might benefit.


Recently emerged swine influenza A virus (H2N3) causes severe pneumonia in Cynomolgus macaques.
The triple reassortant H2N3 virus isolated from diseased pigs in the United States in 2006 is pathogenic for certain mammals without prior adaptation and transmits among swine and ferrets. Adaptation, in the H2 hemagglutinin derived from an avian virus, includes the ability to bind to the mammalian receptor, a significant prerequisite for infection of mammals, in particular humans, which poses a big concern for public health. Here we investigated the pathogenic potential of swine H2N3 in Cynomolgus macaques, a surrogate model for human influenza infection. In contrast to human H2N2 virus, which served as a control and largely caused mild pneumonia similar to seasonal influenza A viruses, the swine H2N3 virus was more pathogenic causing severe pneumonia in nonhuman primates. Both viruses replicated in the entire respiratory tract, but only swine H2N3 could be isolated from lung tissue on day 6 post infection. All animals cleared the infection whereas swine H2N3 infected macaques still presented with pathologic changes indicative of chronic pneumonia at day 14 post infection. Swine H2N3 virus was also detected to significantly higher titers in nasal and oral swabs indicating the potential for animal-to-animal transmission. Plasma levels of IL-6, IL-8, MCP-1 and IFNγ were significantly increased in swine H2N3 compared to human H2N2 infected animals supporting the previously published notion of increased IL-6 levels being a potential marker for severe influenza infections. In conclusion, the swine H2N3 virus represents a threat to humans with the potential for causing a larger outbreak in a non-immune or partially immune population. Furthermore, surveillance efforts in farmed pig populations need to become an integral part of any epidemic and pandemic influenza preparedness.


Raftlike mixtures of sphingomyelin and cholesterol investigated by solid-state 2H NMR spectroscopy.
Sphingomyelin is a lipid that is abundant in the nervous systems of mammals, where it is associated with putative microdomains in cellular membranes and undergoes alterations due to aging or neurodegeneration. We investigated the effect of varying the concentration of cholesterol in binary and ternary mixtures with N-palmitoylsphingomyelin (PSM) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) using deuterium nuclear magnetic resonance ((2)H NMR) spectroscopy in both macroscopically aligned and unoriented multilamellar dispersions. In our experiments, we used PSM and POPC perdeuterated on the N-acyl and sn-1 acyl chains, respectively. By measuring solid-state (2)H NMR spectra of the two lipids separately in mixtures with the same compositions as a function of cholesterol mole fraction and temperature, we obtained clear evidence for the coexistence of two liquid-crystalline domains in distinct regions of the phase diagram. According to our analysis of the first moments M1 and the observed (2)H NMR spectra, one of the domains appears to be a liquid-ordered phase. We applied a mean-torque potential model as an additional tool to calculate the average hydrocarbon thickness, the area per lipid, and structural parameters such as chain extension and thermal expansion coefficient in order to further define the two coexisting phases. Our data imply that phase separation takes place in raftlike ternary PSM/POPC/cholesterol mixtures over a broad temperature range but vanishes at cholesterol concentrations equal to or greater than a mole fraction of 0.33. Cholesterol interacts preferentially with sphingomyelin only at smaller mole fractions, above which a homogeneous liquid-ordered phase is present. The reasons for these phase separation phenomena seem to be differences in the effects of cholesterol on the configurational order of the palmitoyl chains in PSM-d31 and POPC-d31 and a difference in the affinity of cholesterol for sphingomyelin observed at low temperatures. Hydrophobic matching explains the occurrence of raftlike domains in cellular membranes at intermediate cholesterol concentrations but not saturating amounts of cholesterol.


Serum IgG4 as a biomarker reflecting pathophysiology and post-operative recurrence in chronic rhinosinusitis.
Type 2 chronic rhinosinusitis (CRS), especially eosinophilic CRS (ECRS), is an intractable upper airway inflammatory disease. Establishment of serum biomarkers reflecting the pathophysiology of CRS is desirable in a clinical setting. As IgG4 production is regulated by type 2 cytokines, we sought to determine whether serum IgG4 levels can be used as a biomarker for CRS. Association between the serum IgG4 levels and clinicopathological factors was analyzed in 336 CRS patients. Receiver operating characteristics (ROC) analysis was performed to determine the cut-off value of serum IgG4 levels that can be used to predict the post-operative recurrence. Serum IgG4 levels were significantly higher in patients with moderate to severe ECRS versus those with non to mild ECRS. The levels were also significantly higher in asthmatic patients and patients exhibiting recurrence after surgery compared to controls. ROC analysis determined that the best cut-off value for the serum IgG4 level to predict the post-operative recurrence was 95 mg/dL. The corresponding sensitivity and specificity were 39.7% and 80.5%, respectively. When we combined the two cut-off values for the serum IgG4 and periostin, patients with high serum levels of either IgG4 or periostin exhibited a high post-operative recurrence (OR: 3.95) as compared to patients having low serum levels of both IgG4 and periostin. The present results demonstrate that the serum IgG4 level is associated with disease severity and post-operative course in CRS. In particular, the combination of serum IgG4 and periostin could be a novel biomarker that predicts post-operative recurrence.


Eye-Tracking and Corpus-Based Analyses of Syntax-Semantics Interactions in Complement Coercion.
Previous work has shown that the difficulty associated with processing complex semantic expressions is reduced when the critical constituents appear in separate clauses as opposed to when they appear together in the same clause. We investigated this effect further, focusing in particular on complement coercion, in which an event-selecting verb (e.g., began) combines with a complement that represents an entity (e.g., began the memo). Experiment 1 compared reading times for coercion versus control expressions when the critical verb and complement appeared together in a subject-extracted relative clause (SRC) (e.g., The secretary that began/wrote the memo) compared to when they appeared together in a simple sentence. Readers spent more time processing coercion expressions than control expressions, replicating the typical coercion cost. In addition, readers spent less time processing the verb and complement in SRCs than in simple sentences; however, the magnitude of the coercion cost did not depend on sentence structure. In contrast, Experiment 2 showed that the coercion cost was reduced when the complement appeared as the head of an object-extracted relative clause (ORC) (e.g., The memo that the secretary began/wrote) compared to when the constituents appeared together in an SRC. Consistent with the eye-tracking results of Experiment 2, a corpus analysis showed that expressions requiring complement coercion are more frequent when the constituents are separated by the clause boundary of an ORC compared to when they are embedded together within an SRC. The results provide important information about the types of structural configurations that contribute to reduced difficulty with complex semantic expressions, as well as how these processing patterns are reflected in naturally occurring language.


Contrasting effects of phorbol esters on serotonin- and vasopressin-evoked contractions in rat aorta and small mesenteric artery.
Phorbol esters, which activate protein kinase C, modulate vasoconstrictor-induced tension in vascular smooth muscle. We examined the effects of phorbol esters (phorbol 12,13-dibutyrate [PDBu] and 12-O-tetradecanoylphorbol 13-acetate [TPA]) on receptor agonist (serotonin [5-HT] and arginine vasopressin [AVP])-, high K(+)-, and caffeine-induced contractions in rings of rat aorta and a small (second-order) branch of the superior mesenteric artery (SMA). PDBu and TPA significantly augmented agonist-evoked contractions in aorta but diminished those in SMA. For example, 30 nM PDBu increased 5-HT- and AVP-evoked contractions 2.0-2.5-fold in aorta (p less than 0.01) but decreased 5-HT- and AVP-induced contractions by 40-60% in SMA (p less than 0.01). In contrast, PDBu and TPA amplified high K(+)- and 10 mM caffeine-induced contractions in both aorta and SMA. Augmentation of agonist-induced contractions by PDBu was greater in endothelium-denuded aorta than in intact aorta. Two protein kinase C antagonists, H-7 and staurosporine, inhibited 5-HT-evoked contractions in the absence as well as in the presence of PDBu in both types of arteries. The augmentation of contractile responses to caffeine and K+ by phorbol esters in both types of arteries suggests that the phorbols increase the sensitivity of the contractile apparatus to Ca2+, probably by activating protein kinase C. However, the inhibitory effects of phorbols on 5-HT- and AVP-evoked responses in SMA suggest that under these conditions the dominant effect of the phorbols is a marked reduction in the availability of Ca2+ in the SMA but not in the aorta.


Reductive dissolution of supergrowth carbon nanotubes for tougher nanocomposites by reactive coagulation spinning.
Long single-walled carbon nanotubes, with lengths >10 μm, can be spontaneously dissolved by stirring in a sodium naphthalide N,N-dimethylacetamide solution, yielding solutions of individualised nanotubide ions at concentrations up to 0.74 mg mL-1. This process was directly compared to ultrasonication and found to be less damaging while maintaining greater intrinsic length, with increased individualisation, yield, and concentration. Nanotubide solutions were spun into fibres using a new reactive coagulation process, which covalently grafts a poly(vinyl chloride) matrix to the nanotubes directly at the point of fibre formation. The grafting process insulated the nanotubes electrically, significantly enhancing the dielectric constant to 340% of the bulk polymer. For comparison, samples were prepared using both Supergrowth nanotubes and conventional shorter commercial single-walled carbon nanotubes. The resulting nanocomposites showed similar, high loadings (ca. 20 wt%), but the fibres formed with Supergrowth nanotubes showed significantly greater failure strain (up to ∼25%), and hence more than double the toughness (30.8 MJ m-3), compared to composites containing typical ∼1 μm SWCNTs.


The impact of cataract surgery on health related quality of life in Kenya, the Philippines, and Bangladesh.
To assess the impact of cataract surgery on vision related quality of life (VRQoL) and generic health related quality of life (HRQoL) in Kenya, Bangladesh and the Philippines. A multi-center intervention study was conducted. At baseline 651 cases aged ≥50 years with visually impairing cataract (corrected visual acuity (VA) <6/24) and 561 age- gender-matched controls with normal vision (VA>6/18) were interviewed about VRQoL (using the World Health Organization/ Prevention of Blindness and Deafness 20-item Visual Functioning Questionnaire [WHO/PBD VF20]) and generic HRQoL (EuroQol). Cases were offered free/subsidized cataract surgery. Approximately 1 year later participants were re-interviewed. Response rate at follow up was 84% for operated cases and 80% for controls. At baseline, cases had significantly poorer VRQoL scores, were more likely to report problems with the EuroQol 5D five descriptive (EQ-SD) domains (mobility, daily activities, self-care, pain, depression/anxiety) and had significantly poorer self-rated health compared to controls. At follow up VRQoL scores of operated cases improved significantly to approximately equal those of controls. Effect sizes were large (> 0.8) regardless of pre-operative VA. Poor outcome from surgery (VA < 6/60) was associated with smaller VRQoL gains. Among operated cases frequency of reported problems with all the EQ-5D reduced significantly compared to baseline in Kenya and the Philippines, and in mobility, daily activities and self-care in Bangladesh. Self-rated health scores increased significantly in each country. HRQoL of controls remained stable from baseline to follow up. This study among adults undergoing cataract surgery in 3 different low-income settings found evidence of improved VRQoL and generic HRQoL to approximately equal that of controls with normal vision.


Survival of foodborne pathogenic bacteria (Bacillus cereus, Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes) and Bacillus cereus spores in fermented alcoholic beverages (beer and refined rice wine).
Only limited information is available on the microbiological safety of fermented alcoholic beverages because it is still a common belief that such beverages do not provide a favorable environment for bacterial growth and survival. Thus, in this study, we examined the survival of major foodborne pathogens and spores in fermented alcoholic beverages. Foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus) and B. cereus spores (initial population, 3 to 4 log CFU/ml) were inoculated separately into three types of beer and refined rice wine, which were then stored at 5 and 22°C. Bacterial counts were assayed periodically for up to 28 days. Vegetative B. cereus counts decreased rapidly, whereas B. cereus spore counts remained constant (P > 0.05) for a long period of time in all beverages. Vegetative B. cereus cells formed spores in beer at 5 and 22°C, and the spores survived for long periods. Among vegetative cells, E. coli O157:H7 had the highest survival (only 1.49 to 1.56 log reduction during 28 days in beer at 5°C). Beer and refined rice wine supported microbial survival from several days to several weeks. Our results appear to contradict the common belief that pathogens cannot survive in alcoholic beverages. Long-term survival of pathogens (especially B. cereus and E. coli O157:H7) in beer and refined rice wine should be taken into consideration by the manufacturers of these beverages. This study provides basic information that should help further research into microbial survival in alcoholic beverages and increase the microbiological safety regulation of fermented alcoholic beverages.


The development of regret and relief about the outcomes of risky decisions.
Although a number of studies have examined the developmental emergence of counterfactual emotions of regret and relief, none of these has used tasks that resemble those used with adolescents and adults, which typically involve risky decision making. We examined the development of the counterfactual emotions of regret and relief in two experiments using a task in which children chose between one of two gambles that varied in risk. In regret trials they always received the best prize from that gamble but were then shown that they would have obtained a better prize had they chosen the alternative gamble, whereas in relief trials the other prize was worse. We compared two methods of measuring regret and relief based on children's reported emotion on discovering the outcome of the alternative gamble: one in which children judged whether they now felt the same, happier, or sadder on seeing the other prize and one in which children made emotion ratings on a 7-point scale after the other prize was revealed. On both of these methods, we found that 6- and 7-year-olds' and 8- and 9-year-olds' emotions varied appropriately depending on whether the alternative outcome was better or worse than the prize they had actually obtained, although the former method was more sensitive. Our findings indicate that by at least 6 or 7years children experience the same sorts of counterfactual emotions as adults in risky decision-making tasks, and they also suggest that such emotions are best measured by asking children to make comparative emotion judgments.


Promising multifunctional anti-Alzheimer's dimer bis(7)-Cognitin acting as an activator of protein kinase C regulates activities of alpha-secretase and BACE-1 concurrently.
We have recently demonstrated that bis(7)-Cognitin, a promising multifunctional anti-Alzheimer's dimer, can remarkably reduce the generation of amyloid beta peptide (Abeta) by inhibiting beta-secretase (BACE-1) and activating alpha-secretase activity. In this study, the mechanism(s) underlying bis(7)-Cognitin's regulation of the activity of these two proteases was further investigated. In N2a cells stably expressing human amyloid precursor protein with the Swedish mutation (APPswe), the reduction in Abeta production induced by 1microM bis(7)-Cognitin was not altered by the co-pretreatment of muscarinic and nicotinic cholinergic receptor antagonists, indicating that the regulation of APP processing by this dimer is independent of cholinergic transmission. Furthermore, bis(7)-Cognitin (0.1-3microM) significantly increased protein kinase C (PKC) activity in cells and in vitro in a concentration-dependent manner. Administration of a PKC activator, phorbol 12-myristate 13-acetate (PMA), concentration-dependently increased the alpha-secretase cleavage products, and reduced the BACE-1 cleavage products. In addition, the inhibition of PKC prevented PMA- or bis(7)-Cognitin-induced alterations in alpha-secretase and BACE-1 activities, eliminating reductions in Abeta production seen with PMA or the dimer. These results strongly suggest that bis(7)-Cognitin may reduce the biosynthesis of Abeta by inhibiting BACE-1 and activating alpha-secretase concurrently through the direct activation of PKC. Combined with previous findings of direct inhibition of AChE and BACE-1 by this dimer, this work indicates that strategy may have potential to provide new insights into designing novel drugs that target multiple steps of aberrant APP processing to treat Alzheimer's disease.


Characterization of the microRNA pool and the factors affecting its regulatory potential.
The regulation of gene expression by microRNAs (miRNAs) is complex due to a number of variables involved. The potential for one miRNA to target many genes, the presence of multiple miRNA response elements (MREs) in one mRNA molecule and the interplay between RNAs that share common MREs each add a layer of complexity to the process; making it difficult to determine how regulation of gene expression by miRNAs works within the context of the system as a whole. In this study, we used luciferase report vectors inserted with different 3'UTR fragments as probes to detect the repressive effect of the miRNA pool on gene expression and uncovered some essential characteristics of gene regulation mediated by the miRNA pool, such as the nonlinear correlative relationship between the regulatory potential of a miRNA pool and the number of potential MREs, the buffering effect and the saturating effect of the miRNA pool, and the restrictive effect caused by the density of MREs. Through expressing gradient concentration of 3'UTR fragments, we indirectly detected the regulatory potential of the competing endogenous RNA (ceRNA) pool and analysed its effect on the regulatory potential of the miRNA pool. Our results provide some new insights into miRNA pool mediated gene regulation.


Greatly reduced lymphoproliferation in lpr mice lacking major histocompatibility complex class I.
Mice homozygous for the lpr gene have a defect in fas (CD95), a cell surface receptor that belongs to the tumor necrosis factor receptor family and that mediates apoptosis. This genetic abnormality results in lymphoproliferation characterized by the accumulation of CD4-CD8- (double negative [DN]) T cells, autoantibody production, and background strain-dependent, end-organ disease. Our previous results suggested that major histocompatibility complex (MHC) class I may be involved in the development of DN cells. To test this hypothesis, we derived C57BL/6-lpr/lpr (B6/lpr) mice that were deficient for the beta 2-microglobulin gene (beta 2m lpr) and had no detectable class I expression. At 6 mo of age, compared with B6/lpr littermates with normal class I genes, these mice showed greatly reduced lymphadenopathy, mostly due to a dramatic decrease in the number of DN cells. Significant changes in the percentage of other T cell subsets were noted, but only gamma/delta+ T cells showed a marked increase in both percentage and absolute numbers. Analysis of T cell receptor V beta expression of the remaining DN T cells in beta 2m -lpr mice showed a shift to a CD4-like repertoire from a CD8-like repertoire in control B6/lpr mice, indicating that a small MHC class II selected DN population was unmasked in lpr mice lacking class I. We also found that the production of immunoglobulin G (IgG) autoantibodies (antichromatin and anti-single stranded DNA), total IgG and IgG2a, but not total IgM or IgM rheumatoid factor, was significantly reduced in the beta 2m -lpr mice. This work suggests that >90% of DN T cells in lpr mice are derived from the CD8 lineage and are selected on class I. However, a T cell subset selected on class II and T cells expressing gamma/delta are also affected by the lpr defect and become minor components of the aberrant DN population.