General and species-specific recommendations for minimal requirements for the use of cephalopods in scientific research.
Here we list species-specific recommendations for housing, care and management of cephalopod molluscs employed for research purposes with the aim of contributing to the standardization of minimum requirements for establishments, care and accommodation of these animals in compliance with the principles stated in Directive 2010/63/EU. Maximizing their psychophysical welfare was our priority. General recommendations on water surface area, water depth and tank shape here reported represent the outcome of the combined action of the analysis of the available literature and an expertise-based consensus reached - under the aegis of the COST Action FA1301 - among researchers working with the most commonly used cephalopod species in Europe. Information on water supply and quality, environmental conditions, stocking density, feeding and handling are also provided. Through this work we wish to set the stage for a more fertile ground of evidence-based approaches on cephalopod laboratory maintenance, thus facilitating standardization and replicability of research outcomes across laboratories, at the same time maximizing the welfare of these animals.


Evidence of human infection by a new mammarenavirus endemic to Southeastern Asia.
Southeastern Asia is a recognised hotspot for emerging infectious diseases, many of which have an animal origin. Mammarenavirus infections contribute significantly to the human disease burden in both Africa and the Americas, but little data exists for Asia. To date only two mammarenaviruses, the widely spread lymphocytic choriomeningitis virus and the recently described Wēnzhōu virus have been identified in this region, but the zoonotic impact in Asia remains unknown. Here we report the presence of a novel mammarenavirus and of a genetic variant of the Wēnzhōu virus and provide evidence of mammarenavirus-associated human infection in Asia. The association of these viruses with widely distributed mammals of diverse species, commonly found in human dwellings and in peridomestic habitats, illustrates the potential for widespread zoonotic transmission and adds to the known aetiologies of infectious diseases for this region.


Disentangling genetic variation for resistance and tolerance to infectious diseases in animals.
Hosts can in principle employ two different strategies to defend themselves against parasites: resistance and tolerance. Animals typically exhibit considerable genetic variation for resistance (the ability to limit parasite burden). However, little is known about whether animals can evolve tolerance (the ability to limit the damage caused by a given parasite burden). Using rodent malaria in laboratory mice as a model system and the statistical framework developed by plant-pathogen biologists, we demonstrated genetic variation for tolerance, as measured by the extent to which anemia and weight loss increased with increasing parasite burden. Moreover, resistance and tolerance were negatively genetically correlated. These results mean that animals, like plants, can evolve two conceptually different types of defense, a finding that has important implications for the understanding of the epidemiology and evolution of infectious diseases.


The epidermal growth factor receptor family as a central element for cellular signal transduction and diversification.
Homeostasis of multicellular organisms is critically dependent on the correct interpretation of the plethora of signals which cells are exposed to during their lifespan. Various soluble factors regulate the activation state of cellular receptors which are coupled to a complex signal transduction network that ultimately generates signals defining the required biological response. The epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases represents both key regulators of normal cellular development as well as critical players in a variety of pathophysiological phenomena. The aim of this review is to give a broad overview of signal transduction networks that are controlled by the EGFR superfamily of receptors in health and disease and its application for target-selective therapeutic intervention. Since the EGFR and HER2 were recently identified as critical players in the transduction of signals by a variety of cell surface receptors, such as G-protein-coupled receptors and integrins, our special focus is the mechanisms and significance of the interconnectivity between heterologous signalling systems.


Early complications related to approach in thoracic and lumbar spine surgery: a single center prospective study.
Thoracic and lumbar spine surgical procedures are performed for a variety of pathologies. The literature consists of multiple retrospective reviews examining complication prevalence with the surgical treatment of these disorders. However, there is limited direct examination of perioperative complications through a prospective approach. Of the prospective assessments, the majority focuses on specific surgical procedures or provides a limited assessment of certain spinal implants. Prospective assessments of complication incidence in broad patient populations are limited. This article analyzes a prospectively collected database of patients who underwent a thoracic and/or lumbar spine surgery at a large tertiary care center and the effect of surgical approach (anterior or posterior) on the incidence of early complications. Data collection was performed prospectively on 128 patients on the neurosurgical spine service at Thomas Jefferson University hospital from May to December 2008. Data on preoperative diagnosis, medical comorbidities, body mass index, surgical approach and procedure, length of stay, and complication occurrence was recorded and analyzed. Acute complications or adverse events occurring within the initial 30 days after each operative procedure were included. All medical adverse events were included as complications. A previously circumstantiated binary definition of major and minor complications was used to stratify the data. Overall, 76 of 128 patients (59.4%) in this cohort experienced at least one complication. Anterior thoracic and lumbar procedures had an 83.3% (5/6) incidence of complications. Of those patients having solely a posterior thoracic and lumbar procedures, 37 of 75 (49.3%) experienced at least one complication. Combined anterior and posterior surgical procedure had a complication incidence of 34 of 47 (72.3%). The mean number of complications reached significance for the minor and overall complications groups (P = .0076 and .0172, respectively, Poisson regression). Comparing the incidence of complications reveals the overall complications in the posterior alone group compared with the anterior/posterior combined group was significantly lower (P = .0134). Those undergoing instrumented fusions were statistically more likely to encounter complications (P < .001). There is a considerably higher complication incidence than previously reported for thoracic, thoracolumbar, and lumbar spine operations. A prospective approach and a broad definition of perioperative complications increased the recorded incidence of perioperative adverse events and complications. The case complexity of a tertiary referral center may also have escalated the increased incidence. Complications were more common in patients undergoing anterior and anterior/posterior procedures.


Fetal exposure to bisphenol A as a risk factor for the development of childhood asthma: an animal model study.
The prevalence of asthma in industrialized countries has been increasing dramatically and asthma is now the most common chronic disease of children in the United States. The rapidity of the increase strongly suggests that changes in environmental exposures are the likely cause of this epidemic. Further, the early onset of allergic manifestations suggests that these exposures may act on the prenatal development of the immune system. We have focused on the potential effects of bisphenol A (BPA), a chemical pollutant with one of the largest productions, on the development of childhood asthma. We have reported that perinatal BPA exposure promotes the development of allergic asthma in a mouse model. The current study was designed to identify a critical period of BPA exposure and to begin elucidating the mechanisms for this susceptibility. Female BALB/c mice received 10 micro g/ml BPA in their drinking water from one week before pregnancy until the end of the study. Some of the pups were transferred in the first 48 h of life from their BPA-loaded mother to an unexposed mother, or vice versa. Half of the pups were sensitized with a low dose of the experimental allergen ovalbumin (OVA), the rest received PBS as an unsensitized controls. On day 22, the pups were challenged by inhalations of ovalbumin or PBS followed by quantification of eosinophils in and hyperreactivity of their airways, major indicators of experimental asthma in this classical mouse model. Hepatic expression of two isoforms of UDP-glucuronosyltransferase (Ugt) was quantified by quantitative RT-PCR at various ages. Pups exposed to BPA in utero and through breast milk, or in utero only, displayed an asthma phenotype in response to their "suboptimal" allergic sensitization, whereas, pups only exposed to BPA postnatally from breast milk, did not. The expression of Ugt2b1, an isoform related to BPA clearance in rats, was not detectable in mouse fetuses and newborn pups, but increased by day 5 and approached adult levels by day 25. Prenatal exposures that produce environmentally relevant burdens of BPA, followed by postnatal allergic sensitization and challenges, promote the development of experimental allergic asthma. Delayed expression of BPA-metabolizing enzymes may explain, at least in part, the enhanced fetal susceptibility to this common environmental contaminant.


Homo- and hetero-oligomerization of thyrotropin-releasing hormone (TRH) receptor subtypes. Differential regulation of beta-arrestins 1 and 2.
G-protein-coupled receptors (GPCRs) are regulated by a complex network of mechanisms such as oligomerization and internalization. Using the GPCR subtypes for thyrotropin-releasing hormone (TRHR1 and TRHR2), the aim of this study was to determine if subtype-specific differences exist in the trafficking process. If so, we wished to determine the impact of homo- and hetero-oligomerization on TRHR subtype trafficking as a potential mechanism for the differential cellular responses induced by TRH. Expression of either beta-arrestin 1 or 2 promoted TRHR1 internalization. In contrast, only beta-arrestin 2 could enhance TRHR2 internalization. The preference for beta-arrestin 2 by TRHR2 was supported by the impairment of TRHR2 trafficking in mouse embryonic fibroblasts (MEFs) from either a beta-arrestin 2 knockout or a beta-arrestin 1/2 knockout, while TRHR1 trafficking was only abolished in MEFs lacking both beta-arrestins. The differential beta-arrestin-dependence of TRHR2 was directly measured in live cells using bioluminescence resonance energy transfer (BRET). Both BRET and confocal microscopy were also used to demonstrate that TRHR subtypes form hetero-oligomers. In addition, these hetero-oligomers have altered internalization kinetics compared with the homo-oligomer. The formation of TRHR1/2 heteromeric complexes increased the interaction between TRHR2 and beta-arrestin 1. This may be due to conformational differences between TRHR1/2 hetero-oligomers versus TRHR2 homo-oligomers as a mutant TRHR1 (TRHR1 C335Stop) that does not interact with beta-arrestins, could also enhance TRHR2/beta-arrestin 1 interaction. This study demonstrates that TRHR subtypes are differentially regulated by the beta-arrestins and also provides the first evidence that the interactions of TRHRs with beta-arrestin may be altered by hetero-oligomer formation.


Facile synthesis of ultra-small PbSe nanorods for photovoltaic application.
Nanocrystal array solar cells based on lead chalcogenide quantum dots (QDs) have recently achieved a high power conversion efficiency of over 8%. The device performance is expected to further increase by using 1-dimensional nanorods (NRs), due to their improved carrier transport over zero-dimensional quantum dots. However, previously reported PbSe NRs have not been used in solar cells mainly because of their large diameters, resulting in a small bandgap unsuitable for photovoltaic application. In this work, we have demonstrated a new method for synthesizing monodisperse ultra-small PbSe NRs with the diameter approaching 2 nm (Eg > 1.2 eV), which can be attributed to the use of diphenylphosphine (DPP) and trans-2-octenoic acid (t-2-OA). The introduction of trace DPP can greatly lower the reaction temperature, leading to reduced diameters for the obtained PbSe NRs as well as largely increased yield. The use of short-chain t-2-OA together with oleic acid as capping ligands results in high monomer reactivity, fast nucleus diffusion and high growth rate, which realize the anisotropic growth of ultra-small PbSe NRs at low reaction temperatures. The PbSe NRs show n-type properties and high electron mobility as measured using field-effect transistors. The PbSe NRs with narrow diameters also demonstrate a suitable bandgap for photovoltaic application. They are used for the first time in solar cells and their improved efficiency is demonstrated when used together with QDs.


Congenital Toxoplasmosis in Tunisia: Prenatal and Neonatal Diagnosis and Postnatal Follow-up of 35 Cases.
Congenital toxoplasmosis (CT) results from transplacental passage of Toxoplasma gondii to the fetus during acute maternal infection. Our study aims to report clinical and biological patterns of 35 cases of CT diagnosed at the department of the Parasitology of the Pasteur Institute of Tunis and to access the performance of prenatal and early postnatal diagnosis techniques. Serological screening of maternal infection was performed by Immunoglobulin (Ig) M and IgG detection and IgG avidity determination. Prenatal diagnosis was based on both Toxoplasma DNA detection in the amniotic fluid and monthly ultrasound examinations. polymerase chain reaction analysis on amniotic fluid, performed only in 15 cases, detected Toxoplasma's DNA in five cases (33.3%). Ultrasound examination did not reveal any morphological abnormalities. Thirty newborns had serological criteria of Toxoplasma infection. Congenital toxoplasmosis diagnosis was confirmed in 23 cases (76.6%) by immunoblot. Among the 35 born-infants, five (14.3%) were symptomatic: three had chorioretinitis at the first clinical ocular examination, one had neurological symptoms (seizures) with positive parasite DNA in cerebral spinal fluid, and one had both ophthalmological and neurological damages- chorioretinitis and intracranial calcifications in the computed tomography scan. Thirty-four of 35 infected children were treated with pyrimethamine-sulfadiazine combination. Four (11.7%) of the treated infants showed abnormal hematological values because of the treatment side effect. Serological rebound was observed in seven infants. A screening program and a diagnostic algorithm in pregnant women should be implemented in Tunisia to improve the follow-up of seronegative ones and to prevent CT cases.


Prostaglandins A1 and E1 influence gene expression in an established insect cell line (BCIRL-HzAM1 cells).
Prostaglandins (PGs) and other eicosanoids exert important physiological actions in insects and other invertebrates, including influencing ion transport and mediating cellular immune defense functions. Although these actions are very well documented, we have no information on the mechanisms of PGs actions in insect cells. Here we report on the outcomes of experiments designed to test our hypothesis that PGs modulate gene expression in an insect cell line established from pupal ovarian tissue of the moth Helicoverpa zea (BCIRL-HzAM1 cells). We treated cells with either PGA(1) or PGE(1) for 12 or 24h then analyzed cell lysates by 2-D electrophoresis. Analysis of the gels by densitometry revealed substantial changes in protein expression in some of the protein spots we analyzed. These spots were processed for mass spectrometric analysis by MALDI TOF/TOF, which yielded in silico protein identities for all 34 spots. The apparent changes in three of the proteins were confirmed by semi-quantative PCR, showing that the changes in mRNA expression were reflected in changes in protein expression. The 34 proteins were sorted into six categories, protein actions, lipid metabolism, signal transduction, protection, cell functions and metabolism. The findings support the hypothesis that one mechanism of PG action in insect cells is the modulation of gene expression.


Efficient parameter search for qualitative models of regulatory networks using symbolic model checking.
Investigating the relation between the structure and behavior of complex biological networks often involves posing the question if the hypothesized structure of a regulatory network is consistent with the observed behavior, or if a proposed structure can generate a desired behavior. The above questions can be cast into a parameter search problem for qualitative models of regulatory networks. We develop a method based on symbolic model checking that avoids enumerating all possible parametrizations, and show that this method performs well on real biological problems, using the IRMA synthetic network and benchmark datasets. We test the consistency between IRMA and time-series expression profiles, and search for parameter modifications that would make the external control of the system behavior more robust. GNA and the IRMA model are available at http://ibis.inrialpes.fr/.


Adolescents Confusion in Receiving Health Services: A Qualitative Study.
Providing health services for adolescents requires exploration of hidden factors from the perspective of adolescents, providers, and key individuals. Understanding the process of providing health services from adolescents point of view will help receiving and continuation of services. Although many studies have been conducted in Iran on adolescents health needs, few studies have dealt with provision of these services to adolescents. The present study aimed to explain the adolescents and key informants' perception of healthcare provision. The present qualitative study was conducted according to grounded theory. Data were collected using deep semi-structured individual interviews and group discussion. Participants were selected through purposive sampling followed by theoretical sampling. Participants in present study were 65 adolescents, nine youths (19-24-year-old), and 19 parents and key people involved in providing health services. Adolescents and their parents were selected from different parts of Tehran. Data collection continued until data saturation, and was analysed using Corbin-Strauss (2008) method. Issues relating to adolescents perception of the process of providing services included health concerns, society's inappropriate behaviours, and weakness of the health services system in responding to adolescents needs, which as underlying factors contributed to adolescents confusion in receiving services and their proper coping with puberty. Due to lack of education on how to manage puberty by parents, schools, society, and the health system, participating adolescents from Tehran were confused about receiving information and unable to manage puberty problems. Solving this problem requires continuity of services and interaction of family, school and community.


Human frontal lobes are not relatively large.
One of the most pervasive assumptions about human brain evolution is that it involved relative enlargement of the frontal lobes. We show that this assumption is without foundation. Analysis of five independent data sets using correctly scaled measures and phylogenetic methods reveals that the size of human frontal lobes, and of specific frontal regions, is as expected relative to the size of other brain structures. Recent claims for relative enlargement of human frontal white matter volume, and for relative enlargement shared by all great apes, seem to be mistaken. Furthermore, using a recently developed method for detecting shifts in evolutionary rates, we find that the rate of change in relative frontal cortex volume along the phylogenetic branch leading to humans was unremarkable and that other branches showed significantly faster rates of change. Although absolute and proportional frontal region size increased rapidly in humans, this change was tightly correlated with corresponding size increases in other areas and whole brain size, and with decreases in frontal neuron densities. The search for the neural basis of human cognitive uniqueness should therefore focus less on the frontal lobes in isolation and more on distributed neural networks.


Estimating the variation in need for community-based social care by body mass index in England and associated cost: population-based cross-sectional study.
Adult obesity is linked to a greater need for social care because of its association with the development of long term conditions and because obese adults can have physical and social difficulties which inhibit daily living. Obesity thus has considerable social care cost implications but the magnitude of these costs is currently unknown. This paper outlines an approach to estimating obesity-related social care costs in adults aged over 65 in England. We used univariable and multivariable logistic regression models to investigate the relation between the self-reported need for social care and potential determinants, including body mass index (BMI), using data from Health Survey for England. We combined these modelled estimates of need for social care with the mean hours of help received, conditional on receiving any help, to calculate the expected hours of social care received per adult by BMI. BMI is positively associated with self-reported need for social care. A one unit (ie 1 kg/m2) increase in BMI is on average associated with a 5% increase in the odds of need for help with social care (odds ratio 1.05, 95% CI 1.04 to 1.07) in an unadjusted model. Adjusting for long term illness and sociodemographic characteristics we estimate the annual cost of local authority funded care for those who receive it is £599 at a BMI of 23 but £1086 at a BMI of 40. BMI is positively associated with self-reported need for social care after adjustment for sociodemographic factors and limiting long term illness. The increase in need for care with BMI gives rise to additional costs in social care provision which should be borne in mind when calculating the cost-effectiveness of interventions aimed at reducing obesity.


Adolescent olanzapine sensitization is correlated with hippocampal stem cell proliferation in a maternal immune activation rat model of schizophrenia.
Previous work established that repeated olanzapine (OLZ) administration in normal adolescent rats induces a sensitization effect (i.e. increased behavioral responsiveness to drug re-exposure) in the conditioned avoidance response (CAR) model. However, it is unclear whether the same phenomenon can be detected in animal models of schizophrenia. The present study explored the generalizability of OLZ sensitization from healthy animals to a preclinical neuroinflammatory model of schizophrenia in the CAR. Maternal immune activation (MIA) was induced via polyinosinic:polycytidylic acid (PolyI:C) administration into pregnant dams. Behavioral assessments of offspring first identified decreased maternal separation-induced pup ultrasonic vocalizations and increased amphetamine-induced hyperlocomotion in animals prenatally exposed to PolyI:C. In addition, repeated adolescent OLZ administration confirmed the generalizability of the sensitization phenomenon. Using the CAR test, adolescent MIA animals displayed a similar increase in behavioral responsiveness after repeated OLZ exposure during both the repeated drug test days as well as a subsequent challenge test. Neurobiologically, few studies examining the relationship between hippocampal cell proliferation and survival and either antipsychotic exposure or MIA have incorporated concurrent behavioral changes. Thus, the current study also sought to reveal the correlation between OLZ behavioral sensitization in the CAR and hippocampal cell proliferation and survival. 5'-bromodeoxyuridine immunohistochemistry identified a positive correlation between the magnitude of OLZ sensitization (i.e. change in avoidance suppression induced by OLZ across days) and hippocampal cell proliferation. The implications of the relationship between behavioral and neurobiological results are discussed.


Deterministic separation of cancer cells from blood at 10 mL/min.
Circulating tumor cells (CTCs) and circulating clusters of cancer and stromal cells have been identified in the blood of patients with malignant cancer and can be used as a diagnostic for disease severity, assess the efficacy of different treatment strategies and possibly determine the eventual location of metastatic invasions for possible treatment. There is thus a critical need to isolate, propagate and characterize viable CTCs and clusters of cancer cells with their associated stroma cells. Here, we present a microfluidic device for mL/min flow rate, continuous-flow capture of viable CTCs from blood using deterministic lateral displacement (DLD) arrays. We show here that a DLD array device can isolate CTCs from blood with capture efficiency greater than 85% CTCs at volumetric flow rates of up to 10 mL/min with no effect on cell viability.


Bilateral Synchronous Paratesticular Leiomyoma - A Rare Entity.
Paratesticular region is an obscure anatomical region that includes spermatic cord and its contents, tunica of the testes, epididymis and appendices of epididymis and testis. Paratesticular tumours may closely resemble and be clinically indistinguishable from testicular tumours, resulting in a diagnostic dilemma. Preoperative distinction between the benign and malignant paratesticular tumour is extremely difficult, resulting in difficulty in diagnosis and management. We hereby present a case of a 56-year-old male who reported with a painless bilateral scrotal mass and underwent surgical excision of the same. The final biopsy report was paratesticular leiomyoma. The main purpose of this manuscript is to emphasize on the fact that it is imperative for the treating urologists to be aware of such potentially benign rare entity. A high index of clinical suspicion is mandatory, as lack of knowledge about this condition would result in an unnecessary radical orchidectomy.


Human fecal flora: variation in bacterial composition within individuals and a possible effect of emotional stress.
Data are presented on the distribution of 101 bacterial species and subspecies among 1,442 isolates from 25 fecal specimens from three men on: (i) their normal diet and normal living conditions, (ii) normal living conditions but eating the controlled metabolic diet designed for use in the Skylab simulation and missions, and (iii) the Skylab diet in simulated Skylab (isolation) conditions. These bacteria represent the most numerous kinds in the fecal flora. Analyses of the kinds of bacteria from each astronaut during the 5-month period showed more variation in the composition of the flora among the individual astronauts than among the eight or nine samples from each person. This observation indicates that the variations in fecal flora reported previously, but based on the study of only one specimen from each person, more certainly reflect real differences (and not daily variation) in the types of bacteria maintained by individual people. The proportions of the predominant fecal species in the astronauts were similar to those reported earlier from a Japanese-Hawaiian population and were generally insensitive to changes from the normal North American diet to the Skylab diet; only two of the most common species were affected by changes in diet. However, one of the predominant species (Bacteroides fragilis subsp. thetaiotaomicron) appeared to be affected during confinement of the men in the Skylab test chamber. Evidence is presented suggesting that an anger stress situation may have been responsible for the increase of this species simultaneously in all of the subjects studied. Phenotypic characteristics of some of the less common isolates are given. The statistical analyses used in interpretation of the results are discussed.


Measurement of oestrone sulphatase activity in white blood cells to monitor in vivo inhibition of steroid sulphatase activity by oestrone-3-O-sulphamate.
Formation of oestrone via the sulphatase pathway is considered to be a major source of the oestrogen present in breast tumours. Several inhibitors of steroid sulphatase have now been developed for use in the treatment of postmenopausal women with breast cancer. In order to be able to monitor the extent and duration of the inhibition of oestrone sulphatase (E1-STS) readily, we have developed a method to measure the activity of this enzyme in white blood cells (WBCs). Hydrolysis of oestrone sulphate by E1-STS in WBCs was linear with respect to time and the volume of WBCs used. To examine whether the extent of inhibition of E1-STS activity in WBCs, by the inhibitor oestrone-3-O-sulphamate (EMATE), reflected inhibition in other body tissues, activity in WBCs was compared with that in liver and spleen tissue samples from rats. Two hours after an oral dose of EMATE the extent of inhibition of E1-STS detected in WBCs was the same as in the liver. The duration of the inhibition of E1-STS by EMATE, examined over a 1-28 day period in rats, was similar whether monitored in WBCs, liver or spleen. Measurements of E1-STS activity in WBCs were also used to examine the effectiveness of EMATE (0.5 mg/kg) in two male volunteers. E1-STS activity was rapidly inhibited and had only recovered by 27% after 1 month. A marked decrease in the ratio of plasma dehydroepiandrosterone:dehydroepiandrosterone-sulphate (DHA:DHA-S) concentrations was also detected, confirming that EMATE also inhibits DHA-STS activity. The ability to monitor the extent and duration of steroid sulphatase inhibition in WBCs will facilitate the evaluation of this new form of endocrine therapy in women with breast cancer.


Epigenetic down-regulation of death-associated protein kinase in lung cancers.
Death-associated protein kinase (DAPK) is a pro-apoptotic serine/threonine kinase involved in apoptosis. Aberrant methylation of DAPK was reported in lung cancers by methylation-specific PCR. However, we were unable to relate methylation with gene silencing with the same methodology. Our goals were to develop a methodology that related methylation with gene silencing and use it to study the state of the gene in lung cancers. Using a semiquantitative real-time reverse transcription-PCR, DAPK expression was lower in lung cancers than in corresponding nonmalignant bronchial epithelial cells in five of six primary short-term cultures. In continuous cell lines, mRNA expression was down-regulated, as well as compared with nonmalignant bronchial epithelial cells, and its protein was not detected by Western blotting in 17 of 23 (74%) cell lines. We investigated methylation status of 5' flanking region of DAPK by combined bisulfite restriction analysis and bisulfited DNA sequencing. Aberrant methylation was detected in 21 of 48 (44%) cell lines, 2 of 6 primary cultured tumors, and 14 of 38 (37%) primary lung cancers, although varying degrees of methylation were noticed. Furthermore, bisufite sequence data suggested that aberrant methylation might occur selectively at some CpG dinucleotides in cell lines which had absent expression. Treatment with 5-aza-2'-deoxycytidine restored DAPK expression in heavily methylated cell lines tested, and histone deacetylase inhibitor trichostatin A alone restored DAPK expression in some methylated cell lines as well. Our major findings are: (a) DAPK expression is frequently down-regulated in lung cancers; (b) aberrant methylation of DAPK is frequent in lung cancers, although considerable heterogeneity of methylation is present, and some specific CpG dinucleotides are often methylated in expression negative lung cancers; and (c) besides methylation and histone deacetylation, there may be other mechanisms for down-regulation of DAPK expression.


Is Single-Stage Prosthetic Reconstruction Cost Effective? A Cost-Utility Analysis for the Use of Direct-to-Implant Breast Reconstruction Relative to Expander-Implant Reconstruction in Postmastectomy Patients.
Prosthetic breast reconstruction is most commonly performed using the two-stage (expander-implant) technique. However, with the advent of skin-sparing mastectomy and the use of acellular dermal matrices, one-stage prosthetic reconstruction has become more feasible. Prior studies have suggested that one-stage reconstruction has economic advantages relative to two-stage reconstruction despite a higher revision rate. This is the first cost-utility analysis to compare the cost and quality of life of both procedures to guide patient care. A comprehensive literature review was conducted using the MEDLINE, EMBASE, and Cochrane databases to include studies directly comparing matched patient cohorts undergoing single-stage or staged prosthetic reconstruction. Six studies were selected examining 791 direct-to-implant reconstructions and 1142 expander-implant reconstructions. Costs were derived adopting both patient and third-party payer perspectives. Utilities were derived by surveying an expert panel. Probabilities of clinically relevant complications were combined with cost and utility estimates to fit into a decision tree analysis. The overall complication rate was 35 percent for single-stage reconstruction and 34 percent for expander-implant reconstruction. The authors' baseline analysis using Medicare reimbursement revealed a cost decrease of $525.25 and a clinical benefit of 0.89 quality-adjusted life-year when performing single-stage reconstructions, yielding a negative incremental cost-utility ratio. When using national billing, the incremental cost-utility further decreased, indicating that direct-to-implant breast reconstruction was the dominant strategy. Sensitivity analysis confirmed the robustness of the authors' conclusions. Direct-to-implant breast reconstruction is the dominant strategy when used appropriately. Surgeons are encouraged to consider single-stage reconstruction when feasible in properly selected patients.


Control of phospholipid synthesis by phosphorylation of the yeast lipin Pah1p/Smp2p Mg2+-dependent phosphatidate phosphatase.
Phosphorylation of the conserved lipin Pah1p/Smp2p in Saccharomyces cerevisiae was previously shown to control transcription of phospholipid biosynthetic genes and nuclear structure by regulating the amount of membrane present at the nuclear envelope (Santos-Rosa, H., Leung, J., Grimsey, N., Peak-Chew, S., and Siniossoglou, S. (2005) EMBO J. 24, 1931-1941). A recent report identified Pah1p as a Mg2+-dependent phosphatidate (PA) phosphatase that regulates de novo lipid synthesis (Han G.-S., Wu, W. I., and Carman, G. M. (2006) J. Biol. Chem. 281, 9210-9218). In this work we use a combination of mass spectrometry and systematic mutagenesis to identify seven Ser/Thr-Pro motifs within Pah1p that are phosphorylated in vivo. We show that phosphorylation on these sites is required for the efficient transcriptional derepression of key enzymes involved in phospholipid biosynthesis. The phosphorylation-deficient Pah1p exhibits higher PA phosphatase-specific activity than the wild-type Pah1p, indicating that phosphorylation of Pah1p controls PA production. Opi1p is a transcriptional repressor of phospholipid biosynthetic genes, responding to PA levels. Genetic analysis suggests that Pah1p regulates transcription of these genes through both Opi1p-dependent and -independent mechanisms. We also provide evidence that derepression of phospholipid biosynthetic genes is not sufficient to induce the nuclear membrane expansion shown in the pah1delta cells.


Glycine regulation of the N-methyl-D-aspartate receptor-gated ion channel in hippocampal membranes.
The N-methyl-D-aspartate receptor-gated ion channel (NMDA channel) is regulated by glycine. To examine the interaction of glycine and NMDA receptor ligands on NMDA channel function, we used a biochemical marker of channel opening, [3H]N-(1-[thienyl]cyclohexyl)piperidine (TCP). We quantified [3H]glycine,L-[3H]glutamate, and TCP binding in an identical membrane preparation. This allowed direct comparison of NMDA and glycine receptor occupancy and channel activation. Glycine increased the association and dissociation rates of NMDA-dependent TCP binding to hippocampal membranes, without altering the Kd or Bmax for TCP binding. Structurally similar amino acids mimicked the action of glycine, with D-isomers being more potent than L-isomers. The potency of glycine in regulating TCP binding matched that for displacing [3H]glycine. Glycine stimulation of TCP binding required the presence of NMDA agonists and was inhibited by the NMDA antagonist D-2-amino-5-phosphonovaleric acid. Glycine stimulation of NMDA-dependent TCP binding was not associated with an increase in agonist binding to the NMDA receptor. Likewise, NMDA stimulation of glycine-dependent TCP binding was not associated with an increase in the binding of glycine to the glycine receptor. These findings permit the following conclusions: 1) glycine stimulates TCP binding solely by increasing the access of TCP to its site in the NMDA channel; 2) TCP binding can be used to quantify glycine regulation of the NMDA channel; 3) a stereospecific glycine receptor, as part of the NMDA receptor-channel complex, regulates NMDA-evoked channel opening by a mechanism not involving increased agonist binding to the NMDA receptor. Thus, it appears that the mechanism of glycine and NMDA receptor regulation of the NMDA channel is analogous to that of a two-key lock; both receptors, by independent and mutually required mechanisms, alter channel conformation to allow ion passage.


Reliability and validity of a new double poling ergometer for cross-country skiers.
Thirty-eight competitive cross-country skiers were divided into three groups to assess the reliability and validity of a new double poling ergometer. Group A (n = 22) performed two maximal 60-s tests, Group B (n = 8) repeated peak oxygen uptake tests on the double poling ergometer, and Group C (n = 8) performed a maximal 6-min test on the double poling ergometer and a double poling time-trial on snow. The correlation between the power calculated at the flywheel and the power applied at the base of the poles was r = 0.99 (P < 0.05). The power at the poles was 50-70% higher than that at the flywheel. There was a high test-retest reliability in the two 60-s power output tests (coefficient of variation = 3.0%) and no significant difference in peak oxygen uptake in the two 6-min all-out tests (coefficient of variation = 2.4%). There was a strong correlation between the absolute (W) and relative power (W x kg(-1)) output in the 6-min double poling ergometer test and the double poling performance on snow (r = 0.86 and 0.89 respectively; both P < 0.05). In conclusion, our results show that the double poling ergometer has both high reliability and validity. However, the power calculated at the flywheel underestimated the total power produced and needs to be corrected for in ergonomic estimations.


3D shape evolution of microparticles and 3D enabled applications using non-uniform UV flow lithography (NUFL).
The generation of microparticles with non-spherical morphologies has generated extensive interest because of their enhanced physical properties that can increase their performance in a wide variety of clinical and industrial applications. A flow lithographic technique based on stop flow lithography (SFL) recently showed the ability to fabricate particles with 3D shapes via manipulation of the UV intensity profile in a simple 2D microfluidic channel. Here, we further explore this flow lithographic method, called non-uniform flow lithography (NUFL), to investigate the 3D-shape tuning ability for the generation of 3D magnetic microparticles and their potential applications. We characterize the morphological microparticle shape change through variation of polymerization objective, UV intensity, and solution opacity. We also couple the particles' intrinsic anisotropic magnetic properties with an external magnetic field to create chains of bullet- and bell-shaped particles and a valve-like micromachine. In addition, in contrast to other complex and multi-step methodologies, NUFL shows a simple route for the facile creation of 3D microstructure platforms such as microneedles with fully modifiable tip morphology. This method presents intriguing possibilities for growing research within 3D microstructure assembly, micromachine systems and minimally invasive medical interventions.


A school-based harm minimization smoking intervention trial: outcome results.
To determine the impact of a school-based harm minimization smoking intervention compared to traditional abstinence-based approaches. A school-based cluster randomized trial was conducted in Perth, Western Australia in 30 government high schools from 1999 to 2000. Over 4000 students were recruited to participate and schools were assigned randomly to either the harm minimization intervention or a standard abstinence-based programme. The harm minimization intervention comprised eight 1-hour lessons over 2 years, quitting support from school nurses and enactment of policies to support programme components. Comparison schools implemented standard abstinence-based programmes and policies. Cigarette smoking was categorized at two levels: regular smoking, defined as smoking on 4 or more days in the previous week; and 30-day smoking as any smoking within the previous month. At immediate post-test (20 months post-baseline), after accounting for baseline differences, school-level clustering effects, socio-economic status, gender and family smoking, intervention students were less likely to smoke regularly [OR = 0.51, 95% confidence interval (CI) = 0.36, 0.71] or to have smoked within the previous 30 days (OR = 0.69, 95% CI = 0.53, 0.91). The school-based adolescent harm minimization intervention appears to have been more effective than the abstinence-based social influences programme at reducing regular smoking.


Programmable Extreme Chirality in the Visible by Helix-Shaped Metamaterial Platform.
The capability to fully control the chiro-optical properties of metamaterials in the visible range enables a number of applications from integrated photonics to life science. To achieve this goal, a simultaneous control over complex spatial and localized structuring as well as material composition at the nanoscale is required. Here, we demonstrate how circular dichroic bands and optical rotation can be effectively and independently tailored throughout the visible regime as a function of the fundamental meta-atoms properties and of their three dimensional architecture in a the helix-shaped metamaterials. The record chiro-optical effects obtained in the visible range are accompanied by an additional control over optical efficiency, even in the plasmonic context. These achievements pave the way toward fully integrated chiral photonic devices.


A stretch-activated anion channel is up-regulated by the malaria parasite Plasmodium falciparum.
A recent study on malaria-infected human red blood cells (RBCs) has shown induced ion channel activity in the host cell membrane, but the questions of whether they are host- or parasite-derived and their molecular nature have not been resolved. Here we report a comparison of a malaria-induced anion channel with an endogenous anion channel in Plasmodium falciparum-infected human RBCs. Ion channel activity was measured using the whole-cell, cell-attached and excised inside-out configurations of the patch-clamp method. Parasitised RBCs were cultured in vitro, using co-cultured uninfected RBCs as controls. Unstimulated uninfected RBCs possessed negligible numbers of active anion channels. However, anion channels could be activated in the presence of protein kinase A (PKA) and ATP in the pipette solution or by membrane deformation. These channels displayed linear conductance (~15 pS), were blocked by known anion channel inhibitors and showed the permeability sequence I(-) > Br(-) > Cl(-). In addition, in less than 5 % of excised patches, an outwardly rectifying anion channel (~80 pS, outward conductance) was spontaneously active. The host membrane of malaria-infected RBCs possessed spontaneously active anion channel activity, with identical conductances, pharmacology and selectivity to the linear conductance channel measured in stimulated uninfected RBCs. Furthermore, the channels measured in malaria-infected RBCs were shown to have a low open-state probability (P(o)) at positive potentials, which explains the inward rectification of membrane conductance observed when using the whole-cell configuration. The data are consistent with the presence of two endogenous anion channels in human RBCs, of which one (the linear conductance channel) is up-regulated by the malaria parasite P. falciparum.


Polypharmacy correlates with increased risk for hip fracture in the elderly: a population-based study.
Few studies have addressed the association between polypharmacy and hip fracture using population data. We conducted a population-based case-control study to investigate whether polypharmacy increases the risk for hip fracture in the elderly. We used insurance claims data from the Taiwan Bureau of National Health Insurance, a universal insurance program with a coverage rate of more than 98% of the population in Taiwan. We identified 2328 elderly patients with newly diagnosed hip fracture during the period 2005-2007. We randomly selected 9312 individuals without hip fracture to serve as the control group. Patient characteristics, drugs prescribed by physicians, and all types of hip fracture were ascertained. The odds ratio (OR) of hip fracture in association with the number of medications used per day in previous years was assessed.We found that patients were older than controls, predominantly female, and more likely to use 5 or more drugs (22.2% vs. 9.3%, p < 0.0001). The OR of hip fracture increased with the number of medications used per day and with age. Multivariate logistic regression analysis revealed that the overall OR for patients using 10 or more drugs was 8.42 (95% confidence interval [CI], 4.73-15.0) compared with patients who used 0-1 drug per day. However, age-specific analysis revealed that the risk for hip fracture was 23 times greater for patients aged > or = 85 years who used 10 or more drugs than for those aged 65-74 years who used 0-1 drug after controlling for covariates (OR, 23.0; 95% CI, 3.77-140).We conclude that the risk of hip fracture in older people increases with the number of medications used, especially in women. Age interacts with the daily medications for the risk of hip fracture.


Does training or deprivation modulate amygdala activation?
Amygdala involvement in visual emotional processing has been unequivocally established, but the amygdala's participation in auditory emotional processing is less clear. In a previous functional magnetic resonance imaging study (Klinge et al., 2010) we investigated the amygdala's role in auditory emotional processing in blind and sighted humans. We observed stronger amygdala responses to auditory emotional stimuli in the blind who were also better at discriminating emotional stimuli. Importantly, inter-individual differences in this skill correlated with amygdala activation. While these data suggested that the amygdala serves the dominant sensory modality for emotional perception, we could not rule out possible influences of use-dependent training effects. To disambiguate between plastic changes due to deprivation or training we now studied professional actors who have undergone extensive perceptual and expressive auditory emotional trainings but no sensory deprivation. Actors showed emotion discrimination skills comparable to those of blind individuals. However, in contrast to blind volunteers they lacked increased amygdala activations. Surprisingly, actors selectively rated angry stimuli as less intense than control participants, paralleled by a down-regulation of amygdala responses via the anterior cingulate cortex. Taken together, the data from our two studies suggest that enhanced amygdala responses in the blind are mainly due to deprivation-induced plasticity, as highly trained actors who possess the same excellent emotion discrimination skills as the blind did not show these responses. It is also conceivable that the actors' training requires a more professional and controlled dealing with the emotional stimuli, resulting in a down-regulation of affective experience and accompanying amygdala responses.


Is ultrasonography-guided modified coaxial core biopsy of the breast a better technique?
To compare the diagnostic rate, patient comfort, and complications of ultrasonography-guided breast biopsy using a modified coaxial technique with ultrasonography-guided fine needle aspiration and traditional core biopsy. A secondary objective was to describe the use of the coaxial technique for the biopsy of breast lesions and our initial experience. Retrospective study. A regional hospital in Hong Kong. Patients, who were referred for ultrasonography-guided fine needle aspiration or biopsy from 23 November 2007 to 19 March 2008, were divided into three groups. For breast lesions of 8 mm or smaller, fine needle aspirations were performed. For breast lesions larger than 8 mm, the patients were randomly divided into groups receiving traditional core biopsies and coaxial biopsies. The pathological reports were reviewed. Diagnostic rate, patient comfort assessed in terms of pain, and any procedural complications. A total of 45 ultrasonography-guided fine needle aspirations or biopsies of breast lesions were performed. All core biopsies using the traditional core technique (n=15) and coaxial technique (n=16) were diagnostic. While for fine needle aspirations, three (21%) of 14 were not diagnostic and repeat biopsies were undertaken for the corresponding patients. Except for one breast lesion biopsied with the coaxial technique that revealed invasive ductal carcinoma, all others yielded benign lesions. The average pain score for coaxial biopsies was 2.2, while for traditional core biopsies and fine needle aspirations, average scores were 3.7 and 3.8, respectively (P=0.022). No procedure-related complication was documented with either of the three techniques. Modified coaxial core biopsy of the breast has an optimal diagnostic rate and hence avoids the need for repeat biopsies. It is associated with better patient comfort and no increase in the risk of complications.


Enhancing the Enrichment of Pharmacophore-Based Target Prediction for the Polypharmacological Profiles of Drugs.
PharmMapper is a web server for drug target identification by reversed pharmacophore matching the query compound against an annotated pharmacophore model database, which provides a computational polypharmacology prediction approach for drug repurposing and side effect risk evaluation. But due to the inherent nondiscriminative feature of the simple fit scores used for prediction results ranking, the signal/noise ratio of the prediction results is high, posing a challenge for predictive reliability. In this paper, we improved the predictive accuracy of PharmMapper by generating a ligand-target pairwise fit score matrix from profiling all the annotated pharmacophore models against corresponding ligands in the original complex structures that were used to extract these pharmacophore models. The matrix reflects the noise baseline of fit score distribution of the background database, thus enabling estimation of the probability of finding a given target randomly with the calculated ligand-pharmacophore fit score. Two retrospective tests were performed which confirmed that the probability-based ranking score outperformed the simple fit score in terms of identification of both known drug targets and adverse drug reaction related off-targets.


Tissue distribution of opioid receptor gene expression in the rat.
The endogenous opioid peptides have multiple physiological actions at both a central and peripheral level which are mediated by 3 main classes of opioid receptors, mu, delta and kappa. The rat mu, delta and kappa opioid receptors have recently been cloned and their distribution of expression in the central nervous system has been mapped. In these studies we have used the reverse transcriptase polymerase chain reaction and Southern blotting to determine the distribution of expression of the mu, delta and kappa opioid receptors in the peripheral tissues of the rat. All 3 opioid receptors were found to be widely expressed in several peripheral tissues including the small intestine, large intestine, adrenal, kidney, lung, spleen, testis, ovary and uterus. In the stomach, delta and kappa but not mu opioid receptor transcripts were detected. Predominantly delta transcripts were detected in the heart, with no mu and only a weak signal for the kappa receptor. In the liver mu and delta but not kappa transcripts were present. Opioid receptor expression was not detected in endothelium or synovium. There is therefore a broad, but tissue specific distribution of opioid receptor expression in the periphery of the rat, suggesting that the endogenous opioid peptides play an endocrine, paracrine, or autocrine role in the regulation of physiology at a peripheral as well as central level.


Intake and chewing behavior of steers consuming switchgrass preserved as hay or silage.
Effect of preservation method on intake and chewing behavior was examined using a first, late vegetative harvest (mid-June) of Kanlow switchgrass (Panicum virgatum L.). For silage (S), forage was harvested with a commercial field chopper (1.5 to 4 cm average chop length) and ensiled directly in silos 1.2 m in diameter and 3.6 m in height. For hay (H), forage was harvested with a flail-chopper (7 to 15 cm average chop length) and cured as hay in a drier at 77 degrees C. Diets of H and S were fed to six Hereford steers (338+/-5 kg) in a single crossover experiment. Chewing behavior was monitored for 4 d with a computerized system. At feeding, H was higher in DM and contained greater concentrations (DM basis) of NDF, CP, and hemicellulose, but lower concentrations of ADF and cellulose, and had lower in vitro DM disappearance values. Steers fed S had higher intakes of DM (P < .02) and NDF (P < .04) and consumed less water from the water supply (P < .01) than animals fed H. However, total amount of water (from water supply and feed) consumed per kilogram of DMI did not differ between diets. Crude protein intake was similar between diets. Preservation method had no effect on eating time, number of boli ruminated, bolus duration, and number of rumination chews per bolus. Steers fed S made fewer eating chews (P < .10) and ruminated for a longer time (P < .05) while making a greater number of rumination chews (P < .04) than steers fed H. Rumination intercycle time was slightly shorter in steers fed H (P < .05) than in steers fed S. When expressed per kilogram of NDF intake, steers fed S spent less time eating (P < .03) and made fewer eating chews (P < .02) than steers fed H; however, rumination time, number of rumination chews, and number of boli ruminated were not affected by preservation method. Steers fed S ingested feed at a greater rate (P < .03), excreted smaller fecal particles (P < .03), had meals of shorter duration (P < .06), spent less time eating during main meals (meals following feed distribution: P < .05), had more rumination periods (P < .01), and a shorter morning (P < .06) latency time (interval between end of main meal and onset of rumination) than steers fed H. These results indicate that preservation method with its concomitant differences in chop length affected forage chemical composition and voluntary intake, and that differences in chewing behavior occurred mostly during eating.


The FEATURE framework for protein function annotation: modeling new functions, improving performance, and extending to novel applications.
Structural genomics efforts contribute new protein structures that often lack significant sequence and fold similarity to known proteins. Traditional sequence and structure-based methods may not be sufficient to annotate the molecular functions of these structures. Techniques that combine structural and functional modeling can be valuable for functional annotation. FEATURE is a flexible framework for modeling and recognition of functional sites in macromolecular structures. Here, we present an overview of the main components of the FEATURE framework, and describe the recent developments in its use. These include automating training sets selection to increase functional coverage, coupling FEATURE to structural diversity generating methods such as molecular dynamics simulations and loop modeling methods to improve performance, and using FEATURE in large-scale modeling and structure determination efforts.


Retooling Computational Techniques for EEG-Based Neurocognitive Modeling of Children's Data, Validity and Prospects for Learning and Education.
This paper describes continuing research on the building of neurocognitive models of the internal mental and brain processes of children using a novel adapted combination of existing computational approaches and tools, and using electro-encephalographic (EEG) data to validate the models. The guiding working model which was pragmatically selected for investigation was the established and widely used Adaptive Control of Thought-Rational (ACT-R) modeling architecture from cognitive science. The anatomo-functional circuitry covered by ACT-R is validated by MRI-based neuroscience research. The present experimental data was obtained from a cognitive neuropsychology study involving preschool children (aged 4-6), which measured their visual selective attention and word comprehension behaviors. The collection and analysis of Event-Related Potentials (ERPs) from the EEG data allowed for the identification of sources of electrical activity known as dipoles within the cortex, using a combination of computational tools (Independent Component Analysis, FASTICA; EEG-Lab DIPFIT). The results were then used to build neurocognitive models based on Python ACT-R such that the patterns and the timings of the measured EEG could be reproduced as simplified symbolic representations of spikes, built through simplified electric-field simulations. The models simulated ultimately accounted for more than three-quarters of variations spatially and temporally in all electrical potential measurements (fit of model to dipole data expressed as R 2 ranged between 0.75 and 0.98; P < 0.0001). Implications for practical uses of the present work are discussed for learning and educational applications in non-clinical and special needs children's populations, and for the possible use of non-experts (teachers and parents).


Correlates of pedometer use: results from a community-based physical activity intervention trial (10,000 Steps Rockhampton).
Pedometers have become common place in physical activity promotion, yet little information exists on who is using them. The multi-strategy, community-based 10,000 Steps Rockhampton physical activity intervention trial provided an opportunity to examine correlates of pedometer use at the population level. Pedometer use was promoted across all intervention strategies including: local media, pedometer loan schemes through general practice, other health professionals and libraries, direct mail posted to dog owners, walking trail signage, and workplace competitions. Data on pedometer use were collected during the 2-year follow-up telephone interviews from random population samples in Rockhampton, Australia, and a matched comparison community (Mackay). Logistic regression analyses were used to determine the independent influence of interpersonal characteristics and program exposure variables on pedometer use. Data from 2478 participants indicated that 18.1% of Rockhampton and 5.6% of Mackay participants used a pedometer in the previous 18-months. Rockhampton pedometer users (n = 222) were more likely to be female (OR = 1.59, 95% CI: 1.11, 2.23), aged 45 or older (OR = 1.69, 95% CI: 1.16, 2.46) and to have higher levels of education (university degree OR = 4.23, 95% CI: 1.86, 9.6). Respondents with a BMI > 30 were more likely to report using a pedometer (OR = 1.68, 95% CI: 1.11, 2.54) than those in the healthy weight range. Compared with those in full-time paid work, respondents in 'home duties' were significantly less likely to report pedometer use (OR = 0.18, 95% CI: 0.06, 0.53). Exposure to individual program components, in particular seeing 10,000 Steps street signage and walking trails or visiting the website, was also significantly associated with greater pedometer use. Pedometer use varies between population subgroups, and alternate strategies need to be investigated to engage men, people with lower levels of education and those in full-time 'home duties', when using pedometers in community-based physical activity promotion initiatives.


Shotgun nanoLC-MS/MS proteogenomics to document MALDI-TOF biomarkers for screening new members of the Ruegeria genus.
The identification of bacteria by means of matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry directly using whole cells has become a standard method in clinical diagnosis due to its rapidity and simplicity. Nevertheless, the analysis of environmental samples with this approach still represents a challenge due to the enormous microbial diversity existing on earth and the lack of a comprehensive database. Most of the environmentally relevant species comprise only one unique strain, while pathogens such as Escherichia coli, with 667 described strains, are well documented. In such case, identification of the proteins responsible for the peak signals within MALDI-TOF spectra can give crucial information for species discrimination. To give higher confidence in MALDI-TOF biomarker description we exploited information from proteins identified by shotgun nanoLC-MS/MS, consisting of the identification and quantification of low-molecular-weight proteins after SDS-PAGE, in-gel trypsin proteolysis and analysis of tryptic peptides. We also proposed the standardization of the inclusion of internal calibrants in the bacterial sample to improve the accuracy of the MALDI-TOF measurements. In this way, nine candidate biomarkers were tentatively proposed for Ruegeria lacuscaerulensis ITI-1157. The conserved biomarkers were theoretically deduced for all other Ruegeria strains whose genomes have been sequenced and their corresponding m/z MALDI-TOF signals were estimated. Among these, DNA-binding protein, HU, and ribosomal proteins, L29, L30, L32 and S17, were shown experimentally to be also the most prominent and conserved signals in the other strain tested, Ruegeria pomeroyi DSS-3. Thus, we suggested that these five biomarkers, which give rise to 10 m/z peak signals derived from the mono- and doubly protonated proteins, are the best candidates for identifying bacteria belonging to the Ruegeria genus, and quickly assessed their phylogenetic proximity to described species. As an application of these biomarkers, we quickly screened 30 seawater bacterial isolates by MALDI-TOF and found one belonging to the Ruegeria genus, as further confirmed by 16S RNA sequencing. Due to its simplicity and effectiveness, this technique could be of immense value in monitoring bacteria in the environment in the near future.


Mechanical strain induces growth of vascular smooth muscle cells via autocrine action of PDGF.
The effect of cyclic mechanical strain on growth of neonatal rat vascular smooth muscle (VSM) cells were examined. Cells were grown on silicone elastomer plates subjected to cyclic strain (60 cycle/min) by application of a vacuum under the plates. A 48 h exposure to mechanical strain increased the basal rate of thymidine incorporation by threefold and increased cell number by 40% compared with cells grown on stationary rubber plates. Strain also increased the rate of thymidine incorporation in response to alpha-thrombin (from 15- to 33-fold), but not to PDGF. As determined by thymidine autoradiography, strain alone induced a fourfold increase in labeled nuclei at the periphery of dishes, where strain is maximal, and a 2-3-fold increase at the center of dishes. Strain appeared to induce the production of an autocrine growth factor(s), since conditioned medium from cells subjected to strain induced a fourfold increase in DNA synthesis in control cells. Western blots of medium conditioned on the cells subjected to strain indicate that the cells secrete both AA and BB forms of PDGF in response to strain. Northern blots of total cell RNA from cells exposed to strain for 24 h show increased steady-state level of mRNA for PDGF-A. Lastly, polyclonal antibodies to the AA form of PDGF reduced by 75% the mitogenic effect of strain and polyclonal antibodies to AB-PDGF reduced mitogenicity by 50%. Antibodies to bFGF did not significantly reduce the strain-induced thymidine incorporation. Thus, the mechanism of strain-induced growth appears to involve the intermediary action of secreted PDGF.


Confocal microscopy-based goniometry of barnacle cyprid permanent adhesive.
Biological adhesives are materials of particular interest in the fields of bio-inspired technology and antifouling research. The adhesive of adult barnacles has received much attention over the years; however, the permanent adhesive of the cyprid - the colonisation stage of barnacles - is a material about which very little is presently known. We applied confocal laser-scanning microscopy to the measurement of contact angles between the permanent adhesive of barnacle cyprid larvae and self-assembled monolayers of OH- and CH3-terminated thiols. Measurement of contact angles between actual bioadhesives and surfaces has never previously been achieved and the data may provide insight into the physicochemical properties and mechanism of action of these functional materials. The adhesive is a dual-phase system post-secretion, with the behaviour of the components governed separately by the surface chemistry. The findings imply that the cyprid permanent adhesion process is more complex than previously thought, necessitating broad re-evaluation of the system. Improved understanding will have significant implications for the production of barnacle-resistant coatings as well as development of bio-inspired glues for niche applications.


Neuromedin B receptor disruption impairs adipogenesis in mice and 3T3-L1 cells.
Neuromedin B, a bombesin-like peptide, and its receptor, are expressed in white adipose tissue with undefined roles. Female mice with disruption of neuromedin B receptor (NB-R) exhibited partial resistance to diet-induced obesity leading to our hypothesis that NB-R is involved in adipogenesis. Here, we showed that adipose stem/stromal cells (ASC) from perigonadal fat of female NB-R-knockout mice, exposed to a differentiation protocol in vitro, accumulated less lipid (45%) than wild type, suggesting reduced capacity to differentiate under adipogenic input. To further explore mechanisms, preadipocytes 3T3-L1 cells were incubated in the presence of NB-R antagonist (PD168368) during the first 3 days in culture. Cells were analyzed in the end of the treatment (Day 3) and later when fully differentiated (Day 21). NB-R antagonist induced lower number of cells at day 3 and 21 (33-39%), reduced cell proliferation at day 3 (-53%) and reduced lipid accumulation at day 21 (-86%). The mRNA expressions of several adipocyte differentiation markers were importantly reduced at both days: Cebpb and Pparg and Fabp4, Plin-1 and Adipoq, and additionally Lep mRNA at day 21. The antagonist had no effect when incubated with mature 3T3-L1 adipocytes. Therefore, genetically disruption of NB-R in mice ASC or pharmacological antagonism of NB-R in 3T3-L1 cells impairs adipogenesis. The mechanisms suggested by results in 3T3-L1 cells involve reduction of cell proliferation and of early gene expressions, leading to decreased number of mature adipocytes. We speculate that NB-R antagonism may be useful to limit the increase in adiposity due to pre-adipocyte differentiation.


Improved classification of Alzheimer's disease data via removal of nuisance variability.
Diagnosis of Alzheimer's disease is based on the results of neuropsychological tests and available supporting biomarkers such as the results of imaging studies. The results of the tests and the values of biomarkers are dependent on the nuisance features, such as age and gender. In order to improve diagnostic power, the effects of the nuisance features have to be removed from the data. In this paper, four types of interactions between classification features and nuisance features were identified. Three methods were tested to remove these interactions from the classification data. In stratified analysis, a homogeneous subgroup was generated from a training set. Data correction method utilized linear regression model to remove the effects of nuisance features from data. The third method was a combination of these two methods. The methods were tested using all the baseline data from the Alzheimer's Disease Neuroimaging Initiative database in two classification studies: classifying control subjects from Alzheimer's disease patients and discriminating stable and progressive mild cognitive impairment subjects. The results show that both stratified analysis and data correction are able to statistically significantly improve the classification accuracy of several neuropsychological tests and imaging biomarkers. The improvements were especially large for the classification of stable and progressive mild cognitive impairment subjects, where the best improvements observed were 6% units. The data correction method gave better results for imaging biomarkers, whereas stratified analysis worked well with the neuropsychological tests. In conclusion, the study shows that the excess variability caused by nuisance features should be removed from the data to improve the classification accuracy, and therefore, the reliability of diagnosis making.


Risk indicators for dental caries using the International Caries Detection and Assessment System (ICDAS).
While national surveys have found that African-Americans have a higher prevalence and severity of dental caries than white-Americans, there are only a few descriptive studies of the prevalence and severity of dental caries in low-income urban African-Americans. This study assessed the prevalence, severity and determinants of dental caries, using the International Caries Detection and Assessment System (ICDAS). A representative sample of low-income families (a caregiver and a child aged 0-5 years) was selected from low-income census tracts in the city of Detroit, Michigan. Of the 12,655 randomly selected housing units, 10,695 were occupied and 9781 were successfully contacted (91.5%). There were 1386 families with eligible children in the contacted households; and of those, 1021 were interviewed and examined at a permanent examination center organized for this study. This represents an overall response rate of 73.7%. At the center, trained staff interviewed the main caregivers of the selected children, and trained and calibrated dentists examined the caregiver and her/his child. Data used in this study included information gathered from the social, behavioral and parenting questionnaires, the Block Food Frequency Questionnaire (total sugar intake), and data collected from community and census databases. Over 90% of the adults (ages 14-70 years, average 29.3) had at least one noncavitated carious lesion and 82.2% had at least one primary cavitated lesion. Negative binomial regression models found that the age of caregivers and the number of churches in neighborhoods were negatively associated with the number of noncavitated tooth surfaces. Cavitated tooth surfaces were positively associated with age, oral hygiene status, being worried about teeth, a recent visit to a dentist, and the number of grocery stores in the neighborhoods. However, the number of cavitated tooth surfaces was negatively associated with preventive dental visits, positive rating of oral health status and the number of dentists in a community. Dental caries, especially at the noncavitated stage, is highly prevalent in low-income African-American adults in Detroit. A significant increase in the mean number of missing teeth was observed after the age of 34 years. This study found that different individual, social, and community risk indicators were associated with noncavitated versus cavitated tooth surfaces.


Alternative preprocessing of RNA-Sequencing data in The Cancer Genome Atlas leads to improved analysis results.
The Cancer Genome Atlas (TCGA) RNA-Sequencing data are used widely for research. TCGA provides 'Level 3' data, which have been processed using a pipeline specific to that resource. However, we have found using experimentally derived data that this pipeline produces gene-expression values that vary considerably across biological replicates. In addition, some RNA-Sequencing analysis tools require integer-based read counts, which are not provided with the Level 3 data. As an alternative, we have reprocessed the data for 9264 tumor and 741 normal samples across 24 cancer types using the Rsubread package. We have also collated corresponding clinical data for these samples. We provide these data as a community resource. We compared TCGA samples processed using either pipeline and found that the Rsubread pipeline produced fewer zero-expression genes and more consistent expression levels across replicate samples than the TCGA pipeline. Additionally, we used a genomic-signature approach to estimate HER2 (ERBB2) activation status for 662 breast-tumor samples and found that the Rsubread data resulted in stronger predictions of HER2 pathway activity. Finally, we used data from both pipelines to classify 575 lung cancer samples based on histological type. This analysis identified various non-coding RNA that may influence lung-cancer histology. The RNA-Sequencing and clinical data can be downloaded from Gene Expression Omnibus (accession number GSE62944). Scripts and code that were used to process and analyze the data are available from https://github.com/srp33/TCGA_RNASeq_Clinical. firstname.lastname@example.com or firstname.lastname@example.com Supplementary material is available at Bioinformatics online.


Variations in use of PET among Medicare beneficiaries with non-small cell lung cancer, 1998-2007.
To explore demographic and regional factors associated with the use of positron emission tomography (PET) in patients with non-small cell lung cancer (NSCLC) and to determine whether their associations with PET use has changed over time. The Office of Human Research Ethics at the University of North Carolina and the institutional review board of the Duke University Health System approved (with waiver of informed consent) this retrospective analysis of Surveillance Epidemiology and End Results Medicare data for Medicare beneficiaries given a diagnosis of NSCLC between 1998 and 2007. The primary outcome was change in the number of PET examinations 2 months before to 4 months after diagnosis, examined according to year and sociodemographic subgroup. PET use was compared between demographic and geographic subgroups and between early (1998-2000) and late (2005-2007) cohorts by using χ(2) tests. Factors associated with use of PET during the study period were further examined by using logit and linear probability multivariable regression analyses. The final cohort included 46 544 patients with 46 935 cases of NSCLC. By 2005, more than half of patients underwent one or more PET examinations, regardless of demographic subgroup. In multivariable logistic regression analysis, patients who underwent PET were more likely to be married, nonblack, and younger than 80 years and to live in census tracts with higher education levels or in the Northeast (P < .001 for all). Living within 40 miles of a PET facility was initially associated with undergoing PET (P < .001), but this association disappeared by 2007. Imaging rates increased more rapidly in patients who were nonblack (P ≤ .01), patients who were younger than 81 years (P < .001), and patients who lived in the Northeast and South (P < .001). PET imaging among Medicare beneficiaries with NSCLC was initially concentrated among nonblack patients younger than 81 years. Despite widespread adoption among all subgroups, differences within demographic subgroups remained.


Centralised versus decentralised treatment and disposal options for Küçükçekmece Basin of Istanbul city.
Küçükçekmece Lake is a sea lagoon located on the European side of Istanbul city with a drainage area of 4 million inhabitants (year 2040). The original plan to treat all of the wastewater collected from the drainage basin in a single plant located at the Northern end of the lake is compared with the decentralised option involving 4 treatment plants. The initial investment cost of the decentralised option is estimated to be higher than the original plan by an amount of 43 million US $. However, the decentralised option has been chosen finally, considering the low cost and risk of wastewater collection, the possibility of the staging of the services, and the reduced environmental impacts.


Understanding the Role of ApoE Fragments in Alzheimer's Disease.
Alzheimer's disease (AD) is one of the most devastating neurodegenerative diseases. It has been known for decades that the APOE ɛ4 allele is the most significant genetic risk factor for late-onset AD and yet its precise role in the disease remains unclear. The APOE gene encodes apolipoprotein E (apoE), a 35 kDa glycoprotein highly expressed in the brain. There are three different isoforms: apoE3 is the most common allele in the population, whilst apoE2 decreases, and apoE4 increases AD risk. ApoE has numerous functions that affect neuronal and non-neuronal cells, thus how it contributes to disease onset and progression is hotly debated. The apoE4 isoform has been linked to the accumulation of both of the major pathological hallmarks of AD, amyloid plaques containing amyloid β peptides, and neurofibrillary tangles containing hyperphosphorylated tau protein, as well as other hallmarks of the disease, including inflammation and oxidative stress. Numerous studies have shown that apoE undergoes fragmentation in the human brain, and that the fragmentation pattern varies between isoforms. It was previously shown that apoE4 has neurotoxic functions, however recent data has also identified a neuroprotective role for the apoE N-terminal 25 kDa fragment, which is more prevalent in apoE3 individuals. The ability of the apoE 25 kDa fragment to promote neurite outgrowth was recently demonstrated and this suggests there is a potential loss of neuroprotection in apoE4 individuals in addition to the previously described gain of toxic function for specific apoE4 fragments. Here we review the enzymes proposed to be responsible for apoE fragmentation, the specific functions of different apoE fragments and their possible links with AD.


Use of quantitative shotgun proteomics to identify fibronectin 1 as a potential plasma biomarker for clear cell carcinoma of the kidney.
Early detection would be one of the most effective means to improve the outcome of renal cell carcinoma (RCC). We searched for a new plasma marker for RCC using a label-free quantitative shotgun proteomics method. Plasma proteins were digested by trypsin, and the resulting peptides were analyzed by 2-Dimensional Image Converted Analysis of Liquid chromatography mass spectrometry (2DICAL). An identified biomarker candidate was subjected to validation using the Amplified Luminescent Proximity Homogeneous Assay (AlphaLISA). Among a total of 23,407 independent MS peaks, we found that the mean intensity of 59 peaks significantly differed between 20 clear cell RCC patients and 20 healthy controls. MS/MS spectra from 16 of the 59 peaks matched the amino acid sequences of the fibronectin 1 (FN1) gene product. The increased plasma level of FN1 in RCC patients was validated in a cohort of in 77 patients and 130 healthy controls (p < 0.0001). The FN1 is considered to be a promising biomarker candidate for clear cell RCC. Furthermore, AlphaLISA is an alternate to the conventional enzyme-linked immunosorbent assay and should prove useful for the rapid validation of biomarker candidates.


Mitochondrial abnormalities in the putamen in Parkinson's disease dyskinesia.
Prolonged treatment of Parkinson's disease (PD) with levodopa leads to disabling side effects collectively referred to as 'dyskinesias'. We hypothesized that bioenergetic function in the putamen might play a crucial role in the development of dyskinesias. To test this hypothesis, we used post mortem samples of the human putamen and applied real time-PCR approaches and gene expression microarrays. We found that mitochondrial DNA (mtDNA) levels are decreased in patients who have developed dyskinesias, and mtDNA damage is concomitantly increased. These pathologies were not observed in PD subjects without signs of dyskinesias. The group of nuclear mRNA transcripts coding for the proteins of the mitochondrial electron transfer chain was decreased in patients with dyskinesias to a larger extent than in patients who had not developed dyskinesias. To examine whether dopamine fluctuations affect mtDNA levels in dopaminoceptive neurons, rat striatal neurons in culture were repeatedly exposed to levodopa, dopamine or their metabolites. MtDNA levels were reduced after treatment with dopamine, but not after treatment with dopamine metabolites. Levodopa led to an increase in mtDNA levels. We conclude that mitochondrial susceptibility in the putamen plays a role in the development of dyskinesias.


The Influence of Temperature and Leaf Wetness Duration on Infection of Perennial Ryegrass by Rhizoctonia solani.
Controlled environment experiments were conducted to determine the influence of temperature and leaf wetness duration on infection of perennial ryegrass by Rhizoctonia solani. Infection of grass plants raised in pots and exposed to mycelium of R. solani was evaluated at various combinations of temperature and leaf wetness duration. Temperatures included 15, 18, 21, 24, and 27°C. Leaf wetness periods were 9, 12, 15, 18, and 24 h. Disease was most severe (more than 50 leaves with brown patch lesions per pot) when plants were in contact with the inoculum source for 24 h of leaf wetness at 24°C. The least amount of disease (0 leaves with lesions per pot) occurred at 15°C and a 9-h wet period. The data were subjected to analysis of variance with orthogonal polynomial contrasts. Significant effects were included in a regression model that described the response of infection to temperature and wetness duration. The polynomial model included linear and quadratic terms for temperature and wetness duration. The adjusted coefficient of determination for the fitted model was 0.93, indicating an excellent fit to the data. The model is intended for use in an improved brown patch warning system for perennial ryegrass in the midwestern United States.


An atlas of over 90,000 conserved noncoding sequences provides insight into crucifer regulatory regions.
Despite the central importance of noncoding DNA to gene regulation and evolution, understanding of the extent of selection on plant noncoding DNA remains limited compared to that of other organisms. Here we report sequencing of genomes from three Brassicaceae species (Leavenworthia alabamica, Sisymbrium irio and Aethionema arabicum) and their joint analysis with six previously sequenced crucifer genomes. Conservation across orthologous bases suggests that at least 17% of the Arabidopsis thaliana genome is under selection, with nearly one-quarter of the sequence under selection lying outside of coding regions. Much of this sequence can be localized to approximately 90,000 conserved noncoding sequences (CNSs) that show evidence of transcriptional and post-transcriptional regulation. Population genomics analyses of two crucifer species, A. thaliana and Capsella grandiflora, confirm that most of the identified CNSs are evolving under medium to strong purifying selection. Overall, these CNSs highlight both similarities and several key differences between the regulatory DNA of plants and other species.


Long non-coding RNA and Polycomb: an intricate partnership in cancer biology.
High-throughput analyses have revealed that the vast majority of the transcriptome does not code for proteins. These non-translated transcripts, when larger than 200 nucleotides, are termed long non-coding RNAs (lncRNAs), and play fundamental roles in diverse cellular processes. LncRNAs are subject to dynamic chemical modification, adding another layer of complexity to our understanding of the potential roles that lncRNAs play in health and disease. Many lncRNAs regulate transcriptional programs by influencing the epigenetic state through direct interactions with chromatin-modifying proteins. Among these proteins, Polycomb repressive complexes 1 and 2 (PRC1 and PRC2) have been shown to be recruited by lncRNAs to silence target genes. Aberrant expression, deficiency or mutation of both lncRNA and Polycomb have been associated with numerous human diseases, including cancer. In this review, we have highlighted recent findings regarding the concerted mechanism of action of Polycomb group proteins (PcG), acting together with some classically defined lncRNAs including X-inactive specific transcript (XIST), antisense non-coding RNA in the INK4 locus (ANRIL), metastasis associated lung adenocarcinoma transcript 1 (MALAT1), and HOX transcript antisense RNA (HOTAIR).


A Xenopus oocyte beta subunit: evidence for a role in the assembly/expression of voltage-gated calcium channels that is separate from its role as a regulatory subunit.
Two closely related beta subunit mRNAs (xo28 and xo32) were identified in Xenopus oocytes by molecular cloning. One or both appear to be expressed as active proteins, because: (i) injection of Xenopus beta antisense oligonucleotides, but not of sense or unrelated oligonucleotides, significantly reduced endogenous oocyte voltage-gated Ca2+ channel (VGCC) currents and obliterated VGCC currents that arise after injection of mammalian alpha1 cRNAs (alpha(1C) and alpha(1E)); (ii) coinjection of a Xenopus beta antisense oligonucleotide and excess rat beta cRNA rescued expression of alpha1 Ca2+ channel currents; and (iii) coinjection of mammalian alpha1 cRNA with cRNA encoding either of the two Xenopus beta subunits facilitated both activation and inactivation of Ca2+ channel currents by voltage, as happens with most mammalian beta subunits. The Xenopus beta subunit cDNAs (beta3xo cDNAs) predict proteins of 484 aa that differ in only 22 aa and resemble most closely the sequence of the mammalian type 3 beta subunit. We propose that "alpha1 alone" channels are in fact tightly associated alpha1beta3xo channels, and that effects of exogenous beta subunits are due to formation of higher-order [alpha1beta]beta(n) complexes with an unknown contribution of beta3xo. It is thus possible that functional mammalian VGCCs, rather than having subunit composition alpha1beta, are [alpha1beta]beta(n) complexes that associate with alpha2delta and, as appropriate, other tissue-specific accessory proteins. In support of this hypothesis, we discovered that the last 277-aa of alpha(1E) have a beta subunit binding domain. This beta binding domain is distinct from the previously known interaction domain located between repeats I and II of calcium channel alpha1 subunits.


Late Onset of Pancreatic Metastases from Renal Cell Carcinoma. A Case Report.
Metastasis of renal cell carcinoma (RCC) to the pancreas is a rare entity accounting only 0.25-3% of all pancreatic tumors. We present a rare case of isolated three focal pancreatic metastases from RCC, occurring 15 years after the left nephrectomy. The majority of the pancreatic metastases are asymptomatic, as it was in case of our patient excluding the weight loss for the last three months. We demonstrate the importance of the medical history, radiological examinations, histological and immunohistochemical analysis in making a definitive diagnosis.


Encoding of Vicarious Reward Prediction in Anterior Cingulate Cortex and Relationship with Trait Empathy.
Empathy--the capacity to understand and resonate with the experiences of others--can depend on the ability to predict when others are likely to receive rewards. However, although a plethora of research has examined the neural basis of predictions about the likelihood of receiving rewards ourselves, very little is known about the mechanisms that underpin variability in vicarious reward prediction. Human neuroimaging and nonhuman primate studies suggest that a subregion of the anterior cingulate cortex in the gyrus (ACCg) is engaged when others receive rewards. Does the ACCg show specialization for processing predictions about others' rewards and not one's own and does this specialization vary with empathic abilities? We examined hemodynamic responses in the human brain time-locked to cues that were predictive of a high or low probability of a reward either for the subject themselves or another person. We found that the ACCg robustly signaled the likelihood of a reward being delivered to another. In addition, ACCg response significantly covaried with trait emotion contagion, a necessary foundation for empathizing with other individuals. In individuals high in emotion contagion, the ACCg was specialized for processing others' rewards exclusively, but for those low in emotion contagion, this region also responded to information about the subject's own rewards. Our results are the first to show that the ACCg signals probabilistic predictions about rewards for other people and that the substantial individual variability in the degree to which the ACCg is specialized for processing others' rewards is related to trait empathy. Successfully cooperating, competing, or empathizing with others can depend on our ability to predict when others are going to get something rewarding. Although many studies have examined how the brain processes rewards we will get ourselves, very little is known about vicarious reward processing. Here, we show that a subregion of the anterior cingulate cortex in the gyrus (ACCg) shows a degree of specialization for processing others' versus one's own rewards. However, the degree to which the ACCg is specialized varies with people's ability to empathize with others. This new insight into how vicarious rewards are processed in the brain and vary with empathy may be key for understanding disorders of social behavior, including psychopathy and autism.


Tunable plasmonic crystals for edge magnetoplasmons of a two-dimensional electron system.
Plasmonic crystal effects analogous to photonic crystal phenomena such as zone folding and gap opening were observed for edge magnetoplasmons in a two-dimensional electron system with a periodically corrugated boundary at microwave frequencies. Magnetic field dependent photovoltage data provide unequivocal evidence for Bragg reflection. Band gaps up to fifth order were observed. These gaps were investigated as a function of the electron density, the magnetic field, and the periodicity to demonstrate the tunability of the dispersive properties of these plasmonic crystals.


Adrenal gland tumorigenesis after gonadectomy in mice is a complex genetic trait driven by epistatic loci.
Postgonadectomy adrenocortical tumorigenesis is a strain-specific phenomenon in inbred mice, assumed to be caused by elevated LH secretion and subsequent ectopic LH receptor (LHR) overexpression in adrenal gland. However, the molecular mechanisms of this cascade of events remain unknown. In this study, we took advantage of the mouse strain dependency of the phenotype to unravel its genetic basis. Our results present the first genome-wide screening related to this pathology in two independent F2 and backcross populations generated between the neoplastic DBA/2J and the nonsusceptible C57BL/6J strains. Surprisingly, the postgonadectomy elevation of serum LH was followed by similar up-regulation of adrenal LHR expression in both parental strains and their crosses, irrespective of their tumor status, indicating that it is not the immediate cause of the tumorigenesis. Linkage analysis revealed one major significant locus for the tumorigenesis on chromosome 8, modulated by epistasis with another quantitative trait locus on chromosome 18. Weight gain, a secondary phenotype after gonadectomy, showed a significant but separate quantitative trait locus on chromosome 7. Altogether, postgonadectomy adrenocortical tumorigenesis in DBA/2J mice is a dominant trait that is not a direct consequence of adrenal LHR expression but is driven by a complex genetic architecture. Analysis of candidate genes in the tumorigenesis linkage region showed that Sfrp1 (secreted frizzled-related protein 1), a tumor suppressor gene, is differentially expressed in the neoplastic areas. These findings may have relevance to the human pathogenesis of macronodular adrenal hyperplasia and adrenocortical tumors in postmenopausal women and why some of them develop obesity.


The robustness and restoration of a network of ecological networks.
Understanding species' interactions and the robustness of interaction networks to species loss is essential to understand the effects of species' declines and extinctions. In most studies, different types of networks (such as food webs, parasitoid webs, seed dispersal networks, and pollination networks) have been studied separately. We sampled such multiple networks simultaneously in an agroecosystem. We show that the networks varied in their robustness; networks including pollinators appeared to be particularly fragile. We show that, overall, networks did not strongly covary in their robustness, which suggests that ecological restoration (for example, through agri-environment schemes) benefitting one functional group will not inevitably benefit others. Some individual plant species were disproportionately well linked to many other species. This type of information can be used in restoration management, because it identifies the plant taxa that can potentially lead to disproportionate gains in biodiversity.


Artificial intelligence in healthcare: past, present and future.
Artificial intelligence (AI) aims to mimic human cognitive functions. It is bringing a paradigm shift to healthcare, powered by increasing availability of healthcare data and rapid progress of analytics techniques. We survey the current status of AI applications in healthcare and discuss its future. AI can be applied to various types of healthcare data (structured and unstructured). Popular AI techniques include machine learning methods for structured data, such as the classical support vector machine and neural network, and the modern deep learning, as well as natural language processing for unstructured data. Major disease areas that use AI tools include cancer, neurology and cardiology. We then review in more details the AI applications in stroke, in the three major areas of early detection and diagnosis, treatment, as well as outcome prediction and prognosis evaluation. We conclude with discussion about pioneer AI systems, such as IBM Watson, and hurdles for real-life deployment of AI.


The human LFA-3 gene is located at the same chromosome band as the gene for its receptor CD2.
Lymphocyte function-associated antigen 3 (LFA-3) is a widely distributed cell surface glycoprotein that has been assigned a role in cell-cell adhesion on the basis of its capacity to bind to the T-lymphocyte CD2 antigen. The amino acid sequences of the extracellular domains of these two antigens, predicted from their cDNA sequences, show significant similarities, and both are members of the immunoglobulin supergene family. In this communication, a probe prepared from LFA-3 cDNA has been used in Southern blot analyses of somatic cell hybrids and in in situ hybridization to assign the LFA-3 gene to the human chromosome band 1p13. This is the same location previously assigned to CD2. Thus the LFA-3 and CD2 genes have probably arisen by duplication of a common evolutionary precursor. These genes therefore represent a further instance in which related members of the immunoglobulin superfamily are located in adjacent regions of the genome.


Assessment of Enzymatic Browning and Evaluation of Antibrowning Methods on Dates.
Dates' color is known to play a crucial role in determining the value and quality of the fruit. The color changes from the natural accepted golden color to unfavorable dark brown color during storage. In this study, the effect of different color preservation methods (modified atmosphere packaging, cold storage (4°C), sulfur dioxide gas (SO2), and blanching) and its relation to darkening due to action of the browning enzymes and melanin production were investigated. Polyphenol oxidase was shown to be active in all treatments except the samples treated with SO2 gas and steam blanching for ten minutes. Likewise, peroxidase activity showed a similar trend in all samples, but a decrease in activity was observed in sulfated samples and total inactivation in steam blanching for ten minutes. Moreover, sulfated samples have shown improvement in color compared to all other treatments, whereas the steamed samples showed the highest color deterioration. Concurrently, melanin content increased in all samples over the period of storage except in the sulfated samples. FTIR analyses of dates' melanin have revealed similar structural feature to the reference melanin; however, some differences were noticed in the regions 2850-2950 cm-1 and 1690-1705 cm-1 which indicated major structural difference between the two melanin samples. More work is suggested to reveal structural and functional properties of dates' melanin.