Nickel-Catalyzed Carboxylation of Aryl and Heteroaryl Fluorosulfates Using Carbon Dioxide.
The development of efficient and practical methods to construct carboxylic acids using CO2 as a C1 synthon is of great importance. Nickel-catalyzed carboxylation of aryl fluorosulfates and heteroaryl fluorosulfates with CO2 is described, affording arene carboxylic acids with good to excellent yields under mild conditions. In addition, a one-pot phenol fluorosulfation/carboxylation is developed.


Evaluation of children with reading difficulties.
Reading difficulties are common and are associated with poor long-term academic achievement. Evaluation of a child's developmental, educational, and family histories in conjunction with standardized screening tests (e.g., Ages and Stages Questionnaires, Parents' Evaluation of Developmental Status, Safety Word Inventory and Literacy Screener) can increase recognition of risk factors for reading difficulties. Validated, office-based, standardized screening tests and school-administered standardized achievement tests (e.g., California Achievement Tests, Iowa Tests of Basic Skills, Metropolitan Achievement Tests, Stanford Achievement Test) can be used to assess school-age children with reading difficulties. Reading difficulties in children often are caused by environmental and organic risk factors. However, many children have reading or learning disabilities and will have lifelong difficulties with reading despite adequate intervention. Children with substantial reading difficulties should receive a full educational assessment. There is good evidence that individualized instruction emphasizing increased phonologic awareness can have a favorable long-term effect on academic achievement.


Small bifunctional chelators that do not disaggregate amyloid β fibrils exhibit reduced cellular toxicity.
Multifunctional metal chelators that can modulate the amyloid β (Aβ) peptide aggregation and its interaction with metal ions such as copper and zinc hold considerable promise as therapeutic agents for Alzheimer's disease (AD). However, specific rather than systemic metal chelation by these compounds is needed in order to limit any side effects. Reported herein are two novel small bifunctional chelators, 2-[2-hydroxy-4-(diethylamino)phenyl]benzothiazole (L1) and 2-(2-hydroxy-3-methoxyphenyl)benzothiazole (L2), in which the metal-binding donor atoms are integrated within a molecular framework derived from the amyloid-binding fluorescent dye thioflavin T (ThT). The metal-binding properties of L1 and L2 were probed by pH spectrophotometric titrations to determine their pKa values and the corresponding metal complex stability constants, and the isolated metal complexes were structurally characterized. The amyloid-fibril-binding properties of L1 and L2 were investigated by fluorescence titrations and ThT competition assays. Interestingly, L1 and L2 do not lead to the formation of neurotoxic Aβ42 oligomers in the presence or absence of metal ions, as observed by native gel electrophoresis, Western blotting, and transmission electron microscopy. In addition, L1 and L2 were able to reduce the cell toxicity of preformed Aβ42 oligomers and of the copper-stabilized Aβ42 oligomers. Given their ability to reduce the toxicity of soluble Aβ42 and Cu-Aβ42 species, L1 and L2 are promising lead compounds for the development of chemical agents that can control the neurotoxicity of soluble Aβ42 species in AD.


Breadth of participation in organized and unstructured leisure activities over time and rural adolescents' functioning.
Discretionary time outside of school offers a distinct context that can promote adolescent development; however, potential for growth depends in part on how this time is used. In this study, person-centered analyses were used to examine patterns of breadth of participation in both organized and unstructured leisure activities among rural adolescents (N = 276, 49% female) across grades 7, 8, and 10. Adjusting for self-selection factors, the study associated these patterns with 10th grade outcomes. Three profiles of participation emerged: consistently low breadth, consistently average breadth, and consistently high breadth of involvement in both organized and unstructured leisure activities over time. The most popular activity types across profile groups were hanging out with friends, team sports, and outdoor activities. Adolescents involved in a greater breadth of organized activities reported the greatest breadth of involvement in unstructured leisure and the best functioning. Adolescents with low breadth of involvement in both organized and unstructured leisure activities consistently showed poorer outcomes. Adolescents in the high breadth of involvement profile were engaged in all activity types at higher rates than adolescents in the average and low breadth of involvement profiles. We advocate for continued efforts to increase adolescent participation in a variety of different types of out-of-school activities.


Are Anesthesiology Providers Good Guessers? Heart Rate and Oxygen Saturation Estimation in a Simulation Setting.
Anesthesia providers may need to interpret the output of vital sign monitors based on auditory cues, in the context of multitasking in the operating room. This study aims to evaluate the ability of different anesthesia providers to estimate heart rate and oxygen saturation in a simulation setting. Sixty anesthesia providers (residents, nurse anesthetics, and anesthesiologists) were studied. Four scenarios were arranged in a simulation context. Two baseline scenarios with and without waveform visual aid, and two scenarios with variation of heart rate and/or oxygen saturation were used to assess the accuracy of the estimation made by the participants. When the accurate threshold for the heart rate was set at less than 5 beats per minute, the providers only had a correct estimation at two baseline settings with visual aids (p=0.22 and 0.2237). Anesthesia providers tend to underestimate the heart rate when it increases. Providers failed to accurately estimate oxygen saturation with or without visual aid (p=0.0276 and 0.0105, respectively). Change in recording settings significantly affected the accuracy of heart rate estimation (p < 0.0001), and different experience levels affected the estimation accuracy (p=0.041). The ability of anesthesia providers with different levels of experience to assess baseline and variations of heart rate and oxygen saturation is unsatisfactory, especially when oxygen desaturation and bradycardia coexist, and when the subject has less years of experience.


In situ hybridisation detects pro-apoptotic gene expression of a Bcl-2 family member in white syndrome-affected coral.
White syndrome has been described as one of the most prolific diseases on the Great Barrier Reef. Previously, apoptotic cell death has been described as the mechanism driving the characteristic rapid tissue loss associated with this disease, but the molecular mechanisms controlling apoptotic cell death in coral disease have yet to be investigated. In situ methods were used to study the expression patterns of 2 distinct regulators of apoptosis in Acropora hyacinthus tissues undergoing white syndrome and apoptotic cell death. Apoptotic genes within the Bcl-2 family were not localized in apparently healthy coral tissues. However, a Bcl-2 family member (bax-like) was found to localize to cells and tissues affected by white syndrome and those with morphological evidence for apoptosis. A potential up-regulation of pro-apoptotic or bax-like gene expression in tissues with apoptotic cell death adjacent to disease lesions is consistent with apoptosis being the primary cause of rapid tissue loss in coral affected by white syndrome. Pro-apoptotic (bax-like) expression in desmocytes and the basal tissue layer, the calicodermis, distant from the disease lesion suggests that apoptosis may also underlie the sloughing of healthy tissues associated with the characteristic, rapid spread of tissue loss, evident of this disease. This study also shows that in situ hybridisation is an effective tool for studying gene expression in adult corals, and wider application of these methods should allow a better understanding of many aspects of coral biology and disease pathology.


The Use of a "Go/Go" Cognitive Bias Task and Response to a Novel Object to Assess the Effect of Housing Enrichment in Sheep (Ovis aries).
It is widely acknowledged that environmental enrichment can improve animals' welfare and emotional state. This study used cognitive bias and response to a novel object to assess the effect of enriched housing on emotional state in sheep. Eighteen sheep were trained to discriminate between high-quality and low-quality reward locations using a go/go task. Sheep were allocated to a housing treatment (enriched or standard) for three weeks. Judgment bias tests were conducted using three ambiguous, unrewarded locations across three days, followed by assessing responses to a novel object. Effects of anxiety levels shown in training on responses to ambiguous locations and to the presence of a novel object were assessed. Enriched-housed sheep tended to have shorter latencies to approach ambiguous positions than standard-housed sheep (P = 0.08), particularly to the near and middle locations. Sheep from standard housing tended to have shorter latencies to approach food with the novel object present than sheep from enriched hosing (P = 0.06). This study shows that enrichment can affect emotional state and that go/go tasks can be successful in sheep and should be considered in future studies of emotional state.


Force spectroscopy with dual-trap optical tweezers: molecular stiffness measurements and coupled fluctuations analysis.
Dual-trap optical tweezers are often used in high-resolution measurements in single-molecule biophysics. Such measurements can be hindered by the presence of extraneous noise sources, the most prominent of which is the coupling of fluctuations along different spatial directions, which may affect any optical tweezers setup. In this article, we analyze, both from the theoretical and the experimental points of view, the most common source for these couplings in dual-trap optical-tweezers setups: the misalignment of traps and tether. We give criteria to distinguish different kinds of misalignment, to estimate their quantitative relevance and to include them in the data analysis. The experimental data is obtained in a, to our knowledge, novel dual-trap optical-tweezers setup that directly measures forces. In the case in which misalignment is negligible, we provide a method to measure the stiffness of traps and tether based on variance analysis. This method can be seen as a calibration technique valid beyond the linear trap region. Our analysis is then employed to measure the persistence length of dsDNA tethers of three different lengths spanning two orders of magnitude. The effective persistence length of such tethers is shown to decrease with the contour length, in accordance with previous studies.


Mutations in the cadherin superfamily member gene dachsous cause a tissue polarity phenotype by altering frizzled signaling.
The adult cuticular wing of Drosophila is covered by an array of distally pointing hairs that reveals the planar polarity of the wing. We report here that mutations in dachsous disrupt this regular pattern, and do so by affecting frizzled signaling. dachsous encodes a large membrane protein that contains many cadherin domains and dachsous mutations cause deformed body parts. We found that mutations in dachsous also result in a tissue polarity phenotype that at the cellular level is similar to frizzled, dishevelled and prickle, as many cells form a single hair of abnormal polarity. Although their cellular phenotype is similar to frizzled, dishevelled and prickle, dachsous mutant wings display a unique and distinctive abnormal hair polarity pattern including regions of reversed polarity. The development of this pattern requires the function of frizzled pathway genes suggesting that in a dachsous mutant the frizzled pathway is functioning - but in an abnormal way. Genetic experiments indicated that dachsous was not required for the intracellular transduction of the frizzled signal. However, we found that dachsous clones disrupted the polarity of neighboring wild-type cells suggesting the possibility that dachsous affected the intercellular signaling function of frizzled. Consistent with this hypothesis we found that frizzled clones in a dachsous mutant background displayed enhanced domineering non-autonomy, and that the anatomical direction of this domineering non-autonomy was altered in regions of dachsous wings that have abnormal hair polarity. The direction of this domineering nonautonomy was coincident with the direction of the abnormal hair polarity. We conclude that dachsous causes a tissue polarity phenotype because it alters the direction of frizzled signaling.


Distribution and ecological preferences of the freshwater lineage LimA (genus Limnohabitans) revealed by a new double hybridization approach.
The ecological relevance and factors shaping dynamics of Limnohabitans sp. have been largely studied by fluorescence in situ hybridization with a 16S rRNA probe targeting the R-BT group (lineages LimBCDE), but not lineage LimA. Consequently, ecology and distribution of LimA remained unknown. We developed a double hybridization strategy using a novel 23S rRNA probe specifically targeting LimA and LimE that in combination with the existing R-BT probe can discriminate LimA populations. This technique was applied for more than 1000 samples from 46 freshwater sites including long-term data sets from oligo-mesotrophic Lake Zurich, CH and meso-eutrophic Římov reservoir, CZ. LimA was ubiquitously distributed and highly abundant. Observed ecological preferences of LimA in Lake Zurich were in general similar to already reported for Limnohabitans with highest numbers in surface waters during growing seasons. Three times higher densities of LimA were detected in Římov reservoir, where they were significantly more abundant at the riverine zone especially after flood events that introduced fresh terrestrial DOM (dissolved organic matter). Moreover, statistical analyses of biological and physicochemical parameters obtained from small dynamic water bodies confirmed a correspondence between LimA and allochthonous DOM, in opposite to R-BT that was more related to algal primary production.


What are the respective roles of the public and private sectors in pharmaceutical innovation?
What are the respective roles of the public and private sectors in drug development? This question is at the heart of some policy proposals, such as those that would give the government a share of profits from drugs at least partly developed with federal research dollars. This paper provides empirical data on these issues, using information included in the patents on drugs approved between 1988 and 2005. Overall, we find that direct government funding is more important in the development of "priority-review" drugs-sometimes described as the most innovative new drugs-than it is for "standard-review" drugs. Government funding has played an indirect role-for example, by funding basic underlying research that is built on in the drug discovery process-in almost half of the drugs approved and in almost two-thirds of priority-review drugs. Our analyses should help inform thinking about the returns on public research funding-a topic of long-standing interest to economists, policy makers, and health advocates.


Dual root coverage of severely traumatized maxillary central incisors: a divided soft tissue management approach.
A dual mucogingival treatment is presented in which severely traumatized maxillary central incisors sustained lateral and extrusive luxation injuries in a roadside accident. Injuries resulted in complete loss of the overlying radicular labial cortical plate followed by a gingival soft tissue cleft, which created a Grade II Miller recession defect in relation to the labial aspect of the maxillary central incisors. A multidisciplinary treatment approach was used, including emergency care, endodontic therapy, and sequential dual periodontal soft tissue rehabilitation. Subepithelial connective tissue grafting was carried out using two techniques, which proved successful in achieving complete coverage of the defect sites. This two-stage surgical approach enabled achievement of ample vascularization for soft tissue grafting and enhanced its predictability for optimal outcome. At the 3-year follow-up, clinical and radiographic examination showed a maintainable functional and esthetic status.


Cyclooxygenase-inhibiting nitric oxide donators for osteoarthritis.
Nonsteroidal anti-inflammatory drugs (NSAIDs) remain the most commonly used medications for the treatment of the symptoms of many chronic inflammatory diseases, including osteoarthritis. Unfortunately, the toxicity of NSAIDs substantially limits their long-term use. Some newer NSAIDs, namely selective cyclooxygenase (COX)-2 inhibitors, exhibit greater gastrointestinal safety, and concomitant use of anti-secretory drugs can also reduce NSAID-induced gastropathy. However, NSAIDs also adversely affect the cardiovascular system. A new class of anti-inflammatory drugs, COX-inhibiting nitric oxide donators (CINODs), has been designed to exert similar anti-inflammatory effects as NSAIDs, but with an improved safety profile. CINODs release nitric oxide, providing protective effects in the gastrointestinal tract and attenuating the detrimental effects on blood pressure normally associated with NSAIDs. We provide an outline of the rationale for CINODs and their activity, in addition to an overview of the pre-clinical and clinical profile of the most advanced CINOD, naproxcinod.


Domain general mechanisms account for imagined transformations of whole body perspective.
Three experiments investigated the contribution of domain general processes to imagined transformations of whole body perspective. In a task where participants make left-right judgements about a schematic human figure, reaction times made from the point of view of the figure are longer when the figure does not share the same spatial orientation as the participant and are substantially longer than those made from the participant's own point of view. These phenomena have been attributed to a specialised mechanism for imagined perspective transformations. In the present experiments, the effect of orientation on performance was influenced by factors that affect spatial stimulus-response (S-R) compatibility; it was attenuated with reduced dimensional overlap and reversed by crossed S-R mappings. Performance in a control experiment in which no figure was present suggests that processes moderating S-R mappings may account for the effect of adopting another's point of view. Our findings demonstrate a role for domain general processes in imagined transformations of whole body perspective, imply that egocentric codes for spatial relationships within visually presented bodies are automatically generated, and undermine evidence for a specialised mechanism for imagined perspective transformations.


Multicultural Aspects in Functional Gastrointestinal Disorders (FGIDs).
Cross-cultural factors are important in functional gastrointestinal disorders (FGIDs). In the setting of FGIDs, the aims of this review were to: 1) engender interest in global aspects; 2) gain a clearer understanding of culture, race and ethnicity and their effect on patient care and research; 3) facilitate cross-cultural clinical and research competency; and 4) improve and foster the quality and conduct of cross-cultural, multinational research. Cultural variables are inevitably present in the physician-patient context. Food and diets, which differ among cultural groups, are perceived globally as related to or blamed for symptoms. From an individual perspective, biological aspects, such as genetics, the microbiome, environmental hygiene, cytokines and the nervous system, which are affected by cultural differences, are all relevant. Of equal importance are issues related to gender, symptom reporting and interpretation, and family systems. From the physician's viewpoint, understanding the patient's explanatory model of illness, especially in a cultural context, affects patient care and patient education in a multicultural environment. Differences in the definition and use of Complementary and Alternative Medicine and other issues related to healthcare services for the FGIDs are also a relevant cross-cultural issue. This paper highlights the importance of cross-cultural competence in clinical medicine and research.


Osimertinib Induces the Opposite Effect of Proliferation and Migration in the Drug Resistance of EGFR-T790M Non-small Cell Lung Cancer Cells.
Lung cancer has become one of the leading causes of cancer incidence and mortality worldwide. Non-small cell lung carcinoma (NSCLC) is the most common type among all lung cancer cases. NSCLC patients contained high levels of activating epidermal growth factor receptor (EGFR) mutations, such as exon 19 deletion, L858R and T790M. Osimertinib, a third-generation of EGFR tyrosine kinase inhibitor (EGFR-TKI), has therapeutic efficacy on the EGFR-T790M mutation of NSCLC patients; however, treatment of osimertinib still can induce drug resistance in lung cancer patients. Therefore, investigation of the drug resistance mechanisms of osimertinib will provide novel strategies for lung cancer therapy. The H1975OR osimertinib-resistant cell line was established by prolonged exposure with osimertinib derived from the H1975 cells. The cell proliferation ability was evaluated by the cell viability and cell growth assays. The cell migration ability was determined by the Boyden chamber assays. The differential gene expression profile was analyzed by genome-wide RNA sequencing. The protein expression and location were analyzed by western blot and confocal microscopy. In this study, we established the osimertinib-resistant H1975 (T790M/L858R) cancer cells, named the H1975OR cell line. The cell growth ability was decreased in the H1975OR cells by comparison with the H1975 parental cells. Conversely, the cell migration ability was elevated in the H1975OR cells. We found the differential gene expression profile of cell proliferation and migration pathways between the H1975OR and H1975 parental cells. Interestingly, the protein levels of phospho-EGFR, PD-L1, E-cadherin and β-catenin were decreased, but the survivin and N-cadherin proteins were increased in the H1975OR drug-resistant cells. Osimertinib induces the opposite effect of proliferation and migration in the drug resistance of EGFRT790M lung cancer cells. We suggest that differential gene and protein expressions in the cell proliferation and migration pathways may mediate the drug resistance of osimertinib in lung cancer cells. Understanding the molecular drugresistant mechanisms of proliferation and migration pathways of osimertinib may provide novel targets and strategies for the clinical treatment of EGFR-TKIs in lung cancer patients.


Anterior transposition of the superior oblique tendon in the treatment of oculomotor nerve palsy and its influence on postoperative hypertropia.
To determine whether postoperative hypertropia after anterior transposition of the superior oblique tendon without trochleotomy could be avoided with a simplified surgical approach. Eight patients with oculomotor nerve palsy (one patient was bilaterally affected) were retrospectively identified as having undergone anterior transposition of the superior oblique tendon without trochleotomy or vertical rectus muscle surgery between March 1992 and September 1998. The superior oblique tendon was cut at the medial border of the superior rectus muscle and placed 1-3.5 mm anterior to the medial insertion of the superior rectus muscle in each of these patients. Resection of the superior oblique tendon was not performed. The lateral rectus muscle was weakened, and no vertical rectus muscles were resected. Preoperative deviations with the uninvolved eye fixating in primary position ranged from 20-90 prism diopters (delta) of exotropia (mean: 49.3 delta) and from 0-20 delta of hypotropia (mean: 11.25 delta). Postoperative horizontal deviations in the primary gaze position ranged from 12 delta of exotropia to 20 delta of esotropia. Six cases were aligned within 10 delta of exotropia or esotropia. Postoperative vertical deviations in the primary gaze position ranged from 2 delta of hypertropia to 8 delta of hypotropia. Six cases were aligned within 2 delta of deviation. Significant postoperative restrictive hypertropia, or new postoperative paradoxical ocular movements, did not occur in any patient. Patients who underwent follow-up >4 months maintained stable eye alignment. Transposition of the superior oblique tendon without simultaneous resection or trochleotomy, or additional surgery to the vertical rectus muscle simplifies the surgical technique and eliminates subjective decision making regarding the amount of resection.


Roles of ion transport in control of cell motility.
Cell motility is an essential feature of life. It is essential for reproduction, propagation, embryonic development, and healing processes such as wound closure and a successful immune defense. If out of control, cell motility can become life-threatening as, for example, in metastasis or autoimmune diseases. Regardless of whether ciliary/flagellar or amoeboid movement, controlled motility always requires a concerted action of ion channels and transporters, cytoskeletal elements, and signaling cascades. Ion transport across the plasma membrane contributes to cell motility by affecting the membrane potential and voltage-sensitive ion channels, by inducing local volume changes with the help of aquaporins and by modulating cytosolic Ca(2+) and H(+) concentrations. Voltage-sensitive ion channels serve as voltage detectors in electric fields thus enabling galvanotaxis; local swelling facilitates the outgrowth of protrusions at the leading edge while local shrinkage accompanies the retraction of the cell rear; the cytosolic Ca(2+) concentration exerts its main effect on cytoskeletal dynamics via motor proteins such as myosin or dynein; and both, the intracellular and the extracellular H(+) concentration modulate cell migration and adhesion by tuning the activity of enzymes and signaling molecules in the cytosol as well as the activation state of adhesion molecules at the cell surface. In addition to the actual process of ion transport, both, channels and transporters contribute to cell migration by being part of focal adhesion complexes and/or physically interacting with components of the cytoskeleton. The present article provides an overview of how the numerous ion-transport mechanisms contribute to the various modes of cell motility.


The oilseed rape developmental expression resource: a resource for the investigation of gene expression dynamics during the floral transition in oilseed rape.
Transcriptome time series can be used to track the expression of genes during development, allowing the timing, intensity, and dynamics of genetic programmes to be determined. Furthermore, time series analysis can reveal causal relationships between genes, leading to an understanding of how the regulatory networks are rewired during development. Due to its impact on yield, a developmental transition of agricultural interest in crops is the switch from vegetative to floral growth. We previously reported the collection of genome-wide gene expression data during the floral transition in the allopolyploid crop Brassica napus (oilseed rape, OSR). To provide the OSR research community with easy access to this dataset, we have developed the Oilseed Rape Developmental Expression Resource (ORDER; http://order.jic.ac.uk ). ORDER enables users to search for genes of interest and plot expression patterns during the floral transition in both a winter and a spring variety of OSR. We illustrate the utility of ORDER using two case studies: the first investigating the interaction between transcription factors, the second comparing genes that mediate the vernalisation response between OSR and radish (Raphanus sativus L.). All the data is downloadable and the generic website platform underlying ORDER, called AionPlot, is made freely and openly available to facilitate the dissemination of other time series datasets. ORDER provides the OSR research community with access to a dataset focused on a period of OSR development important for yield. AionPlot, the platform on which ORDER is built, will allow researchers from all fields to share similar time series datasets.


Prescription Drug Use and Polypharmacy Among Medicaid-Enrolled Adults with Autism: A Retrospective Cross-Sectional Analysis.
A lack of gold standard treatment for autism spectrum disorders (ASD), no clear ASD management guidelines, and lack of evidence-based pharmacological interventions other than aripiprazole and risperidone elevate the risk of off-label prescribing and adverse effects among individuals with ASD, more so among adults. The aim of this study was to identify and compare the types of prescription drug use, rates of polypharmacy, and characteristics associated with polypharmacy among adults with and without ASD in a retrospective cross-sectional analysis of a three-state Medicaid Analytic eXtract database (2000-2008). Adults aged 22-64 years with ASD (ICD9-CM code: 299.xx) were propensity score-matched to 'no ASD' controls by age, sex, and race. General polypharmacy (≥6 unique classes of prescription drugs in a year) and psychotropic polypharmacy (≥3 unique prescription drug classes of psychotropic medications within a 90-day period) were the main study outcomes. Chi-square tests for rates, t tests for mean number of claims, and multivariate logistic regressions for likelihood of prescription drug use and polypharmacy were run. Annually, almost 75% of adults with ASD had >20 prescription drug claims compared with 33% of adults without ASD. Around 85% of adults with ASD used at least one psychotropic drug class compared with 42% of adults without ASD. Highly common psychotropics were antipsychotics (66%ASD vs 20%noASD), anticonvulsants (59%ASD vs 20%noASD), and anxiolytics/hypnotics/sedatives (21%ASD vs 11%noASD). Other than psychotropics, many adults with ASD used medical prescription drugs such as antimicrobials (47%), dermatologic agents (48%), respiratory agents (38%), gastrointestinal agents (31%), alternative medications (25%), antiparkinsonian agents (22.6%), antihyperlipidemics/statins (7.3%), and immunologics (2.0%). Rates of general (48%ASD vs 32%noASD) and psychotropic polypharmacy (19%ASD vs 6%noASD) were significantly higher in the ASD group. Prescription drug use and polypharmacy rates among adults with ASD are substantially higher than those in an age-, sex-, and race-matched cohort of adults without ASD. Adults with ASD frequently use therapeutic treatments other than psychotropics. Healthcare providers, who usually report low confidence in treating patients with ASD, should play an active role in constant monitoring of prescription drug use patterns and patient response to interventions. Prescribers and caregivers are encouraged to make decisions after weighing the benefits and risks associated with a pharmacological treatment. Further investigations into the common use of any alternative treatments that can affect a patient's response to core treatments should also be conducted.


Pituitary apoplexy after leuprolide injection for ovum donation.
Patients with occult pituitary adenomas infrequently present with pituitary apoplexy. Precipitation of pituitary apoplexy by gonadotropin releasing hormone or gonadotropin releasing hormone agonists has been described. The pathophysiologic mechanism by which these agents induce apoplexy remains unclear. We describe a case of pituitary apoplexy in a young woman receiving leuprolide in preparation for ovum donation. Severe hyponatremia, cerebral vasospasm and infarction, and diabetes insipidus complicated this patient's prolonged hospital course. To our knowledge, pituitary apoplexy after gonadotropin releasing hormone agonist use for ovum donation has not been previously described. The use of leuprolide or other gonadotropin releasing hormone agonists for pituitary down-regulation in conjunction with ovarian stimulation can have serious consequences in women harboring unrecognized pituitary adenomas. Thorough endocrine screening should be performed prior to initiating therapy.


Distinct differences in platelet production and function between neonates and adults: implications for platelet transfusion practice.
Thrombocytopenia is a common problem among sick neonates admitted to the neonatal intensive care unit. Among neonates, preterm infants are the subgroup at highest risk for thrombocytopenia and hemorrhage, which is frequently intracranial. Although there is no evidence of a relationship between platelet (PLT) count and occurrence of major hemorrhage, preterm infants are commonly transfused prophylactically when PLT counts fall below an arbitrary limit, and this threshold is usually higher than for older infants or adults. This liberal practice has been influenced by the observation that, in vitro, neonatal PLTs are hyporeactive in response to multiple agonists. However, full-term infants exhibit normal to increased primary hemostasis due to factors in neonatal blood that enhance the PLT-vessel wall interaction. Additionally, cardiorespiratory problems are considered the main etiologic factors in the development of neonatal intraventricular hemorrhage. In this review, we will discuss the developmental differences that exist in regard to PLT production and function, as well as in primary hemostasis in preterm and term neonates, and the implications of these developmental differences to transfusion medicine. PLT transfusions are not exempt of risk, and a better understanding of the PLT function and the hemostatic profile of premature infants and their changes over time and in response to illness is the starting point to design randomized controlled trials to define optimal use of PLT transfusions in premature neonates. Without these future trials, the marked disparities in PLT transfusion practice in neonates between hospitals and countries will remain over time.


Structure of a uranyl peroxo complex in aqueous solution from first-principles molecular dynamics simulations.
Static ab initio and density-functional computations, as well as Car-Parrinello molecular dynamics simulations in aqueous solution are reported for [UO2(OH)(κ(2)-O2)(H2O)2](-) and [UO2(OH)2(κ(1)-O2H)(H2O)](-). Whereas the κ(1)-hydroperoxo isomer is found to be more stable than the κ(2)-peroxo form in the gas phase, the order of stability is reversed in explicit bulk solution. Based on free energies from thermodynamic integration (BLYP functional), the peroxo form is favoured by ca. 32 kJ mol(-1) in water. This stabilisation is discussed in terms of the hydration shells about the individual ligands and dipole moments of the complexes in water, and highlights the importance of explicit solute-solvent interactions and bulk solvation for the speciation of uranyl(vi) compounds.


Operative mortality and postoperative morbidity of highly selective vagotomy.
In a world-wide survey of the results of 5539 highly selective vagotomies (HSVs) performed electively for duodenal ulcer the operative mortality was found to be 0-3%. This was lower than that found in collected series after either vagotomy with drainage (0-8%) or gastric resection with or without vagotomy (over 1%). Necrosis of the lesser curvature occurred in 10 patients (0-2%) after HSV and caused death in 5(0-1%). Such necrosis is probably ischaemic in origin. Hence reperitonealisation of the raw area on the lesser curvature and prompt laparotomy if the patient develops signs of peritonitis might lower the mortality still further. Three deaths were due to pulmonary embolism, one to mesenteric vascular occlusion, and four to myocardial infarction; such deaths might be reduced by the prophylactic use of low-dose heparin. Persisting gastric stasis requiring drainage occurred in only 0-1% of the patients in the early postoperative period and in 0-6% of the patients later. Hence drainage procedures, which produce side effects such as early dumping, bilious vomiting, and diiarrhoea, could be abandoned if the mean incidence of recurrent ulceration after HSV remains close to its present level. HSV is probably the safest operation for duodenal ulcer because the alimentary tract is not opened and there is no anastomosis, suture line, or stoma.


Cutting edge: T cell-mediated pathology in murine Lyme borreliosis.
Even in the absence of an appropriate model or direct evidence, T cells have been hypothesized to exacerbate the manifestations of Lyme disease. To define definitely the role of T cells in Lyme disease, the course of disease in immunocompetent and B cell-deficient mice was compared. By 8 wk postinoculation, immunocompetent mice resolved both carditis and arthritis, whereas foci of myocarditis and severe destructive arthritis characterized disease of B cell-deficient mice. Cell transfer experiments using infected B6-Rag1 knock out mice demonstrated that: 1) innate immunity mediated the initial sequelae of infection, 2) transferring both naive T cells and B cells induced resolution of carditis and arthritis, 3) infected mice reconstituted with T cells developed myocarditis and severe destructive arthritis, and 4) CD4+ T cells were responsible for the observed immune-mediated pathology. These data demonstrate directly the deleterious effect of T cells in Lyme disease.


Syrinx location and size according to etiology: identification of Chiari-associated syrinx.
OBJECT Syrinx size and location within the spinal cord may differ based on etiology or associated conditions of the brain and spine. These differences have not been clearly defined. METHODS All patients with a syrinx were identified from 14,118 patients undergoing brain or cervical spine imaging at a single institution over an 11-year interval. Syrinx width, length, and location in the spinal cord were recorded. Patients were grouped according to associated brain and spine conditions including Chiari malformation Type I (CM-I), secondary CM (2°CM), Chiari malformation Type 0 (CM-0), tethered cord, other closed dysraphism, and spinal tumors. Syringes not associated with any known brain or spinal cord condition were considered idiopathic. Syrinx characteristics were compared between groups. RESULTS A total of 271 patients with a syrinx were identified. The most common associated condition was CM-I (occurring in 117 patients [43.2%]), followed by spinal dysraphism (20 [7.4%]), tumor (15 [5.5%]), and tethered cord (13 [4.8%]). Eighty-three patients (30.6%) did not have any associated condition of the brain or spinal cord and their syringes were considered idiopathic. Syringes in patients with CM-I were wide (7.8 ± 3.9 mm) compared with idiopathic syringes (3.9 ± 1.0, p < 0.0001) and those associated with tethered cord (4.2 ± 0.9, p < 0.01). When considering CM-I-associated and idiopathic syringes, the authors found that CM-I-associated syringes were more likely to have their cranial extent in the cervical spine (88%), compared with idiopathic syringes (43%; p < 0.0001). The combination of syrinx width greater than 5 mm and cranial extent in the cervical spine had 99% specificity (95% CI 0.92-0.99) for CM-I-associated syrinx. CONCLUSIONS Syrinx morphology differs according to syrinx etiology. The combination of width greater than 5 mm and cranial extent in the cervical spine is highly specific for CM-I-associated syringes. This may have relevance when determining the clinical significance of syringes in patients with low cerebellar tonsil position.


Characterization of a complex chemosensory signal transduction system which controls twitching motility in Pseudomonas aeruginosa.
Virulence of the opportunistic pathogen Pseudomonas aeruginosa involves the coordinate expression of a wide range of virulence factors including type IV pili which are required for colonization of host tissues and are associated with a form of surface translocation termed twitching motility. Twitching motility in P. aeruginosa is controlled by a complex signal transduction pathway which shares many modules in common with chemosensory systems controlling flagella rotation in bacteria and which is composed, in part, of the previously described proteins PilG, PilH, PilI, PilJ and PilK. Here we describe another three components of this pathway: ChpA, ChpB and ChpC, as well as two downstream genes, ChpD and ChpE, which may also be involved. The central component of the pathway, ChpA, possesses nine potential sites of phosphorylation: six histidine-containing phosphotransfer (HPt) domains, two novel serine- and threonine-containing phosphotransfer (SPt, TPt) domains and a CheY-like receiver domain at its C-terminus, and as such represents one of the most complex signalling proteins yet described in nature. We show that the Chp chemosensory system controls twitching motility and type IV pili biogenesis through control of pili assembly and/or retraction as well as expression of the pilin subunit gene pilA. The Chp system is also required for full virulence in a mouse model of acute pneumonia.


The involvement of serotonin metabolism in cigarette smoke-induced oxidative stress in rat lung in vivo.
Recently, we have reported the dysregulation of circulating serotonin (5-hydroxytryptamine, 5-HT) homeostasis in patients with chronic obstructive pulmonary disease (COPD). An increase in metabolism of 5-HT has been reported to induce oxidative stress via monoamine oxidase (MAO)-dependent pathway. The present study aimed at investigating the effect of cigarette smoke exposure on the systemic circulation and local airway 5-HT levels as well as MAO-mediated oxidative pathway using a cigarette smoke-exposed rat model. Male Sprague-Dawley rats (150-200 g) were exposed to either sham air or 4% (v/v, smoke/air) cigarette smoke for 1 hour daily for 56 consecutive days. Sera, bronchoalveolar larvage (BAL) and lung tissues were collected 24 hours after the last exposure. We found a significant reduction in the reduced glutathione (rGSH) and an elevation in advanced oxidation protein products (AOPP), a protein oxidation marker, in the lung of cigarette smoke-exposed group (p < 0.05). A significant increase in 5-HT was found in serum (p < 0.05), but not in the BAL or lung, after cigarette smoke exposure. MAO-A activity was significantly elevated in the lung of cigarette smoke-exposed group (p < 0.05). Furthermore, increased superoxide anion levels were found in lung homogenates of cigarette smoke-exposed rats after incubation with 5-HT (p < 0.05), which was positively associated with the increase in MAO-A activity (r = 0.639, p < 0.05). Our findings supported the presence of GSH disruption and protein oxidation in the lung after cigarette smoke exposure. The metabolism of 5-HT by MAO-A in the lung enhanced cigarette smoke-induced superoxides, which might contribute to the pathogenesis of COPD.


In vitro test of external Qigong.
Practitioners of the alternative medical practice 'external Qigong' generally claim the ability to emit or direct "healing energy" to treat patients. We investigated the ability of experienced Qigong practitioners to enhance the healthy growth of cultured human cells in a series of studies, each following a rigorously designed protocol with randomization, blinding and controls for variability. Qigong practitioners directed healing intentionality toward normal brain cell cultures in a basic science laboratory. Qigong treatments were delivered for 20 minutes from a minimum distance of 10 centimeters. Cell proliferation was measured by a standard colony-forming efficiency (CFE) assay and a CFE ratio (CFE for treated samples/CFE for sham samples) was the dependent measure for each experiment. During a pilot study (8 experiments), a trend of increased cell proliferation in Qigong-treated samples (CFE Qigong/sham ratios > 1.0) was observed (P = 0.162). In a formal study (28 experiments), a similar trend was observed, with Qigong-treated samples showing on average more colony formation than sham samples (P = 0.036). In a replication study (60 experiments), no significant difference between Qigong-treated samples and sham samples was observed (P = 0.465). We observed an apparent increase in the proliferation of cultured cells following external Qigong treatment by practitioners under strictly controlled conditions, but we did not observe this effect in a replication study. These results suggest the need for more controlled and thorough investigation of external Qigong before scientific validation is claimed.


High-throughput droplet-based analysis of influenza A virus genetic reassortment by single-virus RNA sequencing.
The segmented RNA genome of influenza A viruses (IAVs) enables viral evolution through genetic reassortment after multiple IAVs coinfect the same cell, leading to viruses harboring combinations of eight genomic segments from distinct parental viruses. Existing data indicate that reassortant genotypes are not equiprobable; however, the low throughput of available virology techniques does not allow quantitative analysis. Here, we have developed a high-throughput single-cell droplet microfluidic system allowing encapsulation of IAV-infected cells, each cell being infected by a single progeny virion resulting from a coinfection process. Customized barcoded primers for targeted viral RNA sequencing enabled the analysis of 18,422 viral genotypes resulting from coinfection with two circulating human H1N1pdm09 and H3N2 IAVs. Results were highly reproducible, confirmed that genetic reassortment is far from random, and allowed accurate quantification of reassortants including rare events. In total, 159 out of the 254 possible reassortant genotypes were observed but with widely varied prevalence (from 0.038 to 8.45%). In cells where eight segments were detected, all 112 possible pairwise combinations of segments were observed. The inclusion of data from single cells where less than eight segments were detected allowed analysis of pairwise cosegregation between segments with very high confidence. Direct coupling analysis accurately predicted the fraction of pairwise segments and full genotypes. Overall, our results indicate that a large proportion of reassortant genotypes can emerge upon coinfection and be detected over a wide range of frequencies, highlighting the power of our tool for systematic and exhaustive monitoring of the reassortment potential of IAVs.


Effects of fixational saccades on response timing in macaque lateral geniculate nucleus.
Even during active fixation, small eye movements persist that might be expected to interfere with vision. Numerous brain mechanisms probably contribute to discounting this jitter. Changes in the timing of responses in the visual thalamus associated with fixational saccades are considered in this study. Activity of single neurons in alert monkey lateral geniculate nucleus (LGN) was recorded during fixation while pseudorandom visual noise stimuli were presented. The position of the stimulus on the display monitor was adjusted based on eye position measurements to control for changes in retinal locations due to eye movements. A method for extracting nonstationary first-order response mechanisms was applied, so that changes around the times of saccades could be observed. Saccade-related changes were seen in both amplitude and timing of geniculate responses. Amplitudes were greatly reduced around saccades. Timing was retarded slightly during a window of about 200 ms around saccades. That is, responses became more sustained. These effects were found in both parvocellular and magnocellular neurons. Timing changes in LGN might play a role in maintaining cortical responses to visual stimuli in the presence of eye movements, compensating for the spatial shifts caused by saccades via these shifts in timing.


Long form of latent TGF-β binding protein 1 (Ltbp1L) regulates cardiac valve development.
Transforming Growth Factor β (TGF-β) is crucial for valve development and homeostasis. The long form of Latent TGF-β binding protein 1 (LTBP1L) covalently binds all TGF-β isoforms and regulates their bioavailability. Ltbp1L expression analysis during valvulogenesis revealed two patterns of Ltbp1L production: an early one (E9.5-11.5) associated with endothelial-to-mesenchymal transformation (EMT); and a late one (E12.5 to birth) contemporaneous with valve remodeling. Similarly, histological analysis of Ltbp1L(-/-) developing valves identified two different pathologies: generation of hypoplastic endocardial cushions in early valvulogenesis, followed by development of hyperplastic valves in late valvulogenesis. Ltbp1L promotes valve EMT, as Ltbp1L absence yields hypoplastic endocardial cushions in vivo and attenuated EMT in vitro. Ltbp1L(-/-) valve hyperplasia in late valvuogenesis represents a consequence of prolonged EMT. We demonstrate that Ltbp1L is a major regulator of Tgf-β activity during valvulogenesis since its absence results in a perturbed Tgf-β pathway that causes all Ltbp1L(-/-) valvular defects.


In Situ Derivatization and Quantification of Seven Carbonyls in Cigarette Mainstream Smoke.
Carbonyls, especially aldehydes, are a group of harmful volatile organic compounds that are found in tobacco smoke. Seven carbonyls are listed on the FDA's harmful and potential harmful constituents list for tobacco or tobacco smoke. Carbonyls have reactive functional groups and thus are challenging to quantitatively measure in cigarette smoke. The traditional method of measuring carbonyls in smoke involves solvent-filled impinger trapping and derivatization. This procedure is labor-intensive and generates significant volumes of hazardous waste. We have developed a new method to efficiently derivatize and trap carbonyls from mainstream smoke in situ on Cambridge filter pads. The derivatized carbonyls are extracted from the pads and subsequently quantified by ultra-high-pressure liquid chromatography coupled with tandem mass spectrometry. The new method has been validated and applied to research and commercial cigarettes. Carbonyl yields from research cigarettes are comparable to those from other published literature data. With a convenient smoke collection apparatus, a 4 min sample analysis time, and a low- or submicrogram detection limit, this new method not only simplifies and speeds the detection of an important class of chemical constituents in mainstream smoke but also reduces reactive losses and provides a more accurate assessment of carbonyl levels in smoke. Excellent accuracy (average 98%) and precision (14% average relative standard deviation in research cigarettes) ensure this new method's sufficient fidelity to characterize conventional combusted tobacco products, with potential application toward new or emerging products.


Sex is not everything: the role of gender in early performance of a fundamental laparoscopic skill.
Existing literature on the acquisition of surgical skills suggests that women generally perform worse than men. This literature is limited by looking at an arbitrary number of trials and not adjusting for potential confounders. The objective of this study was to evaluate the impact of gender on the learning curve for a fundamental laparoscopic task. Thirty-two medical students performed the FLS peg transfer task and their scores were plotted to generate a learning curve. Nonlinear regression was used to estimate learning plateau and learning rate. Variables that may affect performance were assessed using a questionnaire. Innate visual-spatial abilities were evaluated using tests for spatial orientation, spatial scanning, and perceptual abilities. Score on first peg transfer attempt, learning plateau, and learning rate were compared for men and women using Student's t test. Innate abilities were correlated to simulator performance using Pearson's coefficient. Multivariate linear regression was used to investigate the effect of gender on early laparoscopic performance after adjusting for factors found significant on univariate analysis. Statistical significance was defined as P < 0.05. Nineteen men and 13 women participated in the study; 30 were right-handed, 12 reported high interest in surgery, and 26 had video game experience. There were no differences between men and women in initial peg transfer score, learning plateau, or learning rate. Initial peg transfer score and learning rate were higher in subjects who reported having a high interest in surgery (P = 0.02, P = 0.03). Initial score also correlated with perceptual ability score (P = 0.03). In multivariate analysis, only surgical interest remained a significant predictor of score on first peg transfer (P = 0.03) and learning rate (P = 0.02), while gender had no significant relationship to early performance. Gender did not affect the learning curve for a fundamental laparoscopic task, while interest in surgery and perceptual abilities did influence early performance.


Synthesis and Characterization of Double Solid Solution (Zr,Ti)2(Al,Sn)C MAX Phase Ceramics.
Quasi phase-pure (>98 wt %) MAX phase solid solution ceramics with the (Zr,Ti)2(Al0.5,Sn0.5)C stoichiometry and variable Zr/Ti ratios were synthesized by both reactive hot pressing and pressureless sintering of ZrH2, TiH2, Al, Sn, and C powder mixtures. The influence of the different processing parameters, such as applied pressure and sintering atmosphere, on phase purity and microstructure of the produced ceramics was investigated. The addition of Sn to the (Zr,Ti)2AlC system was the key to achieve phase purity. Its effect on the crystal structure of a 211-type MAX phase was assessed by calculating the distortions of the octahedral M6C and trigonal M6A prisms due to steric effects. The M6A prismatic distortion values were found to be smaller in Sn-containing double solid solutions than in the (Zr,Ti)2AlC MAX phases. The coefficients of thermal expansion along the ⟨ a⟩ and ⟨ c⟩ directions were measured by means of Rietveld refinement of high-temperature synchrotron X-ray diffraction data of (Zr1- x,Ti x)2(Al0.5,Sn0.5)C MAX phase solid solutions with x = 0, 0.3, 0.7, and 1. The thermal expansion coefficient data of the Ti2(Al0.5,Sn0.5)C solid solution were compared with those of the Ti2AlC and Ti2SnC ternary compounds. The thermal expansion anisotropy increased in the (Zr,Ti)2(Al0.5,Sn0.5)C double solid solution MAX phases as compared to the Zr2(Al0.5,Sn0.5)C and Ti2(Al0.5,Sn0.5)C end-members.


Characterization and turnover of CD73/IP(3)R3-positive microvillar cells in the adult mouse olfactory epithelium.
The main olfactory epithelium consists of 4 major cell types: sensory neurons, supporting cells, microvillar cells, and basal progenitor cells. Several populations of microvillar olfactory cells have been described, whose properties are not yet fully understood. In this study, we aimed to clarify the classification of microvillar cells by introducing a specific marker, CD73. Furthermore, we investigated the turnover of CD73-microvillar cells during adult life. Using direct and indirect immunofluorescence in adult main olfactory epithelium, we first demonstrate that ecto-5'-nucleotidase (CD73) is a reliable marker for microvillar cells reported previously to express phospholipase C β2 (PLC β2) along with type 3 IP(3) receptors (IP(3)R3) and transient receptor potential channels 6 (TRPC6), as well as for cells labeled by transgenic expression of tauGFP driven by the IP(3)R3 promoter. The ubiquitous CD73 immunoreactivity in the microvilli of these 2 cell populations indicates that they correspond to the same cell type (CD73-microvillar cell), endowed with a signal transduction cascade mobilizing Ca(++) from intracellular stores. These microvillar cells respond to odors, possess a basal process, and do not degenerate after bulbectomy, suggesting that they contribute to cellular homeostasis in the olfactory epithelium. Next, we examined whether CD73-microvillar cells undergo turnover in the adult olfactory epithelium. By combining CD73 immunofluorescence and BrdU pulse labeling, we show delayed BrdU incorporation in a small fraction of CD73-positive microvillar cells, which persists for several weeks after BrdU administration. These findings indicate that CD73-microvillar cells likely differentiate from proliferating progenitor cells and have a slow turnover despite their apical position in the olfactory epithelium. These combined properties are unique among olfactory cells, in line with the possibility that they might regulate cellular homeostasis driven by extracellular ATP and adenosine.


The PKU mouse project: its history, potential and implications.
To produce genetic-based animal models for the human disease phenylketonuria (PKU), we treated mice with the powerful germline mutagen ethylnitrosourea and screened the progeny of these animals for the symptom hyperphenylalaninemia (HPH). Six independent mutant strains have been produced to date that exhibit heritable HPH. The first mutation isolated was found to cause a reduced level of GTP-cyclohydrolase I activity and, as such, yields a model for tetrahydrobiopterin-dependent HPH. The next two mutations have yet to be fully characterized but cause syndromes that appear distinct from any PKU or HPH syndromes yet reported for humans and they are allelic. Next we isolated a mutation that caused a marked reduction in hepatic phenylalanine hydroxylase activity levels. The enzyme deficiency was not sufficient to cause a PKU syndrome but instead produced a mild HPH syndrome. This strain played an instrumental role, however, in the identification of two additional mutant strains that appear to model human PKU very accurately in the laboratory mouse. These latter strains have levels of HPH very similar to human PKU patients, exhibit a phenylalanine-dependent hypopigmentation, and have reproductive difficulties that resemble human maternal PKU.


High water mobility on the ice-binding surface of a hyperactive antifreeze protein.
Antifreeze proteins (AFPs) prevent uncontrolled ice formation in organisms exposed to subzero temperatures by binding irreversibly to specific planes of nascent ice crystals. To understand the thermodynamic driving forces and kinetic mechanism of AFP activity, it is necessary to characterize the hydration behavior of these proteins in solution. With this aim, we have studied the hyperactive insect AFP from Tenebrio molitor (TmAFP) with the (17)O magnetic relaxation dispersion (MRD) method, which selectively monitors the rotational motion and exchange kinetics of water molecules on picosecond-microsecond time scales. The global hydration behavior of TmAFP is found to be similar to non-antifreeze proteins, with no evidence of ice-like or long-ranged modifications of the solvent. However, two sets of structural water molecules, located within the core and on the ice-binding face in the crystal structure of TmAFP, may have functional significance. We find that 2 of the 5 internal water molecules exchange with a residence time of 8 +/- 1 micros at 300 K and a large activation energy of approximately 50 kJ mol(-1), reflecting intermittent large-scale conformational fluctuations in this exceptionally dense and rigid protein. Six water molecules arrayed with ice-like spacing in the central trough on the ice-binding face exchange with bulk water on a sub-nanosecond time scale. The combination of high order and fast exchange may allow these water molecules to contribute entropically to the ice-binding affinity without limiting the absorption rate.


NPC1 Deficiency in Mice is Associated with Fetal Growth Restriction, Neonatal Lethality and Abnormal Lung Pathology.
The rare lysosomal storage disorder Niemann-Pick disease type C1 (NPC1) arises from mutation of NPC1, which encodes a lysosomal transmembrane protein essential for normal transport and trafficking of cholesterol and sphingolipids. NPC1 is highly heterogeneous in both clinical phenotypes and age of onset. Previous studies have reported sub-Mendelian survival rates for mice homozygous for various Npc1 mutant alleles but have not studied the potential mechanisms underlying this phenotype. We performed the first developmental analysis of a Npc1 mouse model, Npc1em1Pav, and discovered significant fetal growth restriction in homozygous mutants beginning at E16.5. Npc1em1Pav/em1Pav mice also exhibited cyanosis, increased respiratory effort, and over 50% lethality at birth. Analysis of neonatal lung tissues revealed lipid accumulation, notable abnormalities in surfactant, and enlarged alveolar macrophages, suggesting that lung abnormalities may be associated with neonatal lethality in Npc1em1Pav/em1Pav mice. The phenotypic severity of the Npc1em1Pav model facilitated this first analysis of perinatal lethality and lung pathology in an NPC1 model organism, and this model may serve as a useful resource for developing treatments for respiratory complications seen in NPC1 patients.


Alteration of sensory neurons and spinal response to an experimental osteoarthritis pain model.
To verify the biologic links between progressive cellular and structural alterations within knee joint components and development of symptomatic chronic pain that are characteristic of osteoarthritis (OA), and to investigate the molecular basis of alterations in nociceptive pathways caused by OA-induced pain. An animal model of knee joint OA pain was generated by intraarticular injection of mono-iodoacetate (MIA) in Sprague-Dawley rats, and symptomatic pain behavior tests were performed. Relationships between development of OA with accompanying pain responses and gradual alterations in cellular and structural knee joint components (i.e., cartilage, synovium, meniscus, subchondral bone) were examined by histologic and immunohistologic analysis, microscopic examination, and microfocal computed tomography. Progressive changes in the dynamic interrelationships between peripheral knee joint tissue and central components of nociceptive pathways caused by OA-induced pain were examined by investigating cytokine production and expression in sensory neurons of the dorsal root ganglion and spinal cord. We observed that structural changes in components of the peripheral knee joint correlate with alterations in the central compartments (dorsal root ganglia and the spinal cord) and symptomatic pain assessed by behavioral hyperalgesia. Our comparative gene expression studies revealed that the pain pathways in MIA-induced knee OA may overlap, at least in part, with neuropathic pain mechanisms. Similar results were also observed upon destabilization of the knee joint in the anterior cruciate ligament transection and destabilization of the medial meniscus models of OA. Our results indicate that MIA-induced joint degeneration in rats generates an animal model that is suitable for mechanistic and pharmacologic studies on nociceptive pain pathways caused by OA, and provide key in vivo evidence that OA pain is caused by central sensitization through communication between peripheral OA nociceptors and the central sensory system. Furthermore, our data suggest a mechanistic overlap between OA-induced pain and neuropathic pain.


Go/no-go decision in anaesthesia: wide variation in risk tolerance amongst anaesthetists.
The variability in risk tolerance in medicine is not well understood. Parallels are often drawn between aviation and anaesthesia. The aviation industry is perceived as culturally risk averse, and part of preflight checks involves a decision on whether the flight can operate. This is sometimes termed a go/no-go decision. This questionnaire study was undertaken to explore the equivalent go/no-go decision in anaesthesia. We presented anaesthetists with a range of situations in which additional risk might be expected and asked them to decide whether they would proceed with the case. An electronic questionnaire was distributed to anaesthetic colleagues of all grades in one National Health Service Trust. Eleven scenarios, all drawn from critical incident data, were presented. Participants were invited to consider whether they would proceed, how they would modify their anaesthetic technique, and to predict whether a colleague with similar experience would make the same decision. Textual responses were analysed qualitatively. The scenario response rate was 28%. Consultants were significantly more likely to proceed than trainees. In no scenario was there absolute agreement over whether to proceed, even in scenarios where national guidelines would suggest a case should be cancelled. Thematic analysis suggested a wide variability in what anaesthetists consider acceptable or professional behaviour. It is clear that safety decisions cannot be made in isolation and that clinicians must consider operational requirements, such as throughput, when making a go/no-go decision. The level of variability in decision-making was surprising, particularly for scenarios that appeared to go against guidelines.


Anchored p90 ribosomal S6 kinase 3 is required for cardiac myocyte hypertrophy.
Cardiac myocyte hypertrophy is the main compensatory response to chronic stress on the heart. p90 ribosomal S6 kinase (RSK) family members are effectors for extracellular signal-regulated kinases that induce myocyte growth. Although increased RSK activity has been observed in stressed myocytes, the functions of individual RSK family members have remained poorly defined, despite being potential therapeutic targets for cardiac disease. To demonstrate that type 3 RSK (RSK3) is required for cardiac myocyte hypertrophy. RSK3 contains a unique N-terminal domain that is not conserved in other RSK family members. We show that this domain mediates the regulated binding of RSK3 to the muscle A-kinase anchoring protein scaffold, defining a novel kinase anchoring event. Disruption of both RSK3 expression using RNA interference and RSK3 anchoring using a competing muscle A-kinase anchoring protein peptide inhibited the hypertrophy of cultured myocytes. In vivo, RSK3 gene deletion in the mouse attenuated the concentric myocyte hypertrophy induced by pressure overload and catecholamine infusion. Taken together, these data demonstrate that anchored RSK3 transduces signals that modulate pathologic myocyte growth. Targeting of signaling complexes that contain select kinase isoforms should provide an approach for the specific inhibition of cardiac myocyte hypertrophy and for the development of novel strategies for the prevention and treatment of heart failure.


Peritonsillar Abscess Model for Ultrasound Diagnosis Using Inexpensive Materials.
This low-cost peritonsillar abscess model is intended for the education of emergency medicine and otolaryngology residents and advanced care practitioners of all training levels. With incidence rates as high as 124 per 100,000 in the 14-21 age range, peritonsillar abscesses (PTA) are one of the more common head and neck soft tissue infections encountered in the emergency department.1 Peritonsillar abscesses can present to the emergency department in critically ill patients with the dangers of airway compromise and further local spreading. Emergency medicine (EM) residents need practice to properly identify and to minimize procedural complications such as perforation of nearby vessels, aspiration pneumonitis, and airway compromise. A major tool used in the emergency department that can help prevent complications is the use of ultrasound, which the Accreditation Council for Graduate Medical Education (ACGME) requires residents to become proficient at.2 Historically, computed tomography (CT) scanning to diagnose along with blind drainage has been the method of choice. With a sensitivity of 95.2%, intraoral ultrasound can minimize both radiation and procedure related complications.3 The current simulators available come at significant capital expenditure and do not provide high-fidelity ultrasound experience. Here we design and implement a low-cost trainer for residents to use ultrasound to diagnose and drain a PTA. By the end of this instructional session learners should be able to: 1) identify and discuss the indications, contraindications, and complications associated with peritonsillar abscesses, 2) properly identify and measure a PTA through ultrasound, and 3) competently perform ultrasound-guided peritonsillar abscess drainage on a simulator and remove fluid. This PTA model utilizes task trainers designed from Styrofoam wig heads. An airway was modeled using readily available wood shop tools and balloons filled with a fluid mixture containing coconut lotion, water, and fragrance beads, which were inserted into the airway. This unique mixture within the balloons creates a realistic echogenicity of an abscess with loculations. With emergency medicine clinical faculty guidance and the use of ultrasound, learners are able to identify a peritonsillar abscess and subsequently demonstrate drainage of fluid with a needle and syringe. This PTA model was tested with a group of 36 emergency medicine residents. Optional, anonymous post surveys were completed by 17 residents. A 5-point Likert Scale was used to assess utility of this model. The majority of users agreed the model provides a realistic image of the disease for diagnosis by ultrasound with a score of 3.6 and felt more comfortable identifying and draining peritonsillar abscesses with scores of 3.7 and 3.6 respectively. Learners' surveys revealed the session was useful and improved their knowledge with both scoring 3.8. No critical feedback was given by learners or instructors. The efficacy of the content was assessed by evaluators observing proper ultrasound, procedure set up, and drainage of PTA. This inexpensive model to expose residents to proper PTA drainage was effective considering learners' high response to post-procedure survey scales. The results of our pilot implementation showed this model has utility in teaching ultrasound guided identification and drainage of PTA's. With minimal build and optimized instruction time, we can improve residents' comfort in performing this procedure and allow for important simulation experience in a safe, controlled environment. Simulation, emergency medicine, peritonsillar abscess, otolaryngology.


A no-go theorem for one-layer feedforward networks.
It is often hypothesized that a crucial role for recurrent connections in the brain is to constrain the set of possible response patterns, thereby shaping the neural code. This implies the existence of neural codes that cannot arise solely from feedforward processing. We set out to find such codes in the context of one-layer feedforward networks and identified a large class of combinatorial codes that indeed cannot be shaped by the feedforward architecture alone. However, these codes are difficult to distinguish from codes that share the same sets of maximal activity patterns in the presence of subtractive noise. When we coarsened the notion of combinatorial neural code to keep track of only maximal patterns, we found the surprising result that all such codes can in fact be realized by one-layer feedforward networks. This suggests that recurrent or many-layer feedforward architectures are not necessary for shaping the (coarse) combinatorial features of neural codes. In particular, it is not possible to infer a computational role for recurrent connections from the combinatorics of neural response patterns alone. Our proofs use mathematical tools from classical combinatorial topology, such as the nerve lemma and the existence of an inverse nerve. An unexpected corollary of our main result is that any prescribed (finite) homotopy type can be realized by a subset of the form [Formula: see text], where Ρ is a polyhedron.


Maternal pertussis is hazardous for a newborn: a case report.
Pertussis, or whooping cough, a highly contagious disease caused by Bordetella pertussis, is making a comeback globally and nationally in spite of reasonable vaccination coverage. Worldwide, there have been increasing reports of Bordetella pertussis infection among adolescents and adults, but the peak incidence and highest mortality occur among infants. We report a 19-day-old female infant presenting with progressive respiratory failure. The mother was the only familial contact who complained of mild cough. However, occasional apneic episodes with cyanosis and peripheral lymphocytosis prompted us to examine the presence of Bordetella pertussis, which remains a significant cause of morbidity and mortality in unimmunized infants. Understanding the source of pertussis transmission to infants may provide new approaches to prevent pertussis in the most vulnerable infants. Various potential strategies have been reviewed or recommended in countries with the aim of better protecting infants against pertussis. Public health measures to prevent the disease could be strengthened and booster vaccinations against pertussis considered.


A comparison of mail-service and retail community pharmacy claims in 5 prescription benefit plans.
Little evidence has been presented to date that would either support or refute a widely held belief that mail-service pharmacy utilization routinely produces savings in drug benefit costs for prescription benefit plan sponsors. To present a comparative analysis of mail-service and community pharmacy service drug benefit costs for 5 employer-sponsored prescription drug benefit plans. A cross-sectional comparison of 17,725 matched transaction pairs of community and mail-service prescriptions from a data set comprised 484,987 prescription claims from a convenience sample of 5 employer-sponsored prescription benefit plans. Differences between community pharmacy and mail-service prescription transactions were examined at the per-unit level of analysis for drug ingredient costs, dispensing fees, co-payments, dollar amounts paid by plan sponsor, and total dollar amounts. Overall, the total cost of prescriptions was lower through mail-service pharmacies for all 5 plans studied. Two of 5 plans had co-payment incentives to use mail-service, yet plan sponsors paid more for mail-service drugs; respectively, 4.5% and 8.3% more overall, 25.0% and 21.4% more for generic medications; and 3.0% and 7.0% more for brand name medications. Mail-service co-payments were 48.9% and 51.7% lower. Mail-service utilization rates were 15.2% and 31.5% of the total number of prescriptions dispensed in the period studied. Three of 5 plans had no co-payment incentive to use mail-service and paid less for mail-service drugs; respectively, 18.7%, 6.6%, and 15.7% less overall; 17.4%, 15.6%, and 7.9% less for generic medications; and 18.8%, 5.2%, and 16.6% less for brand name medications. Mail-service co-payments were 10.5% more, 5.2% less, and 1.8% more than community pharmacy co-payments, respectively. Mail-service utilization rates were 0.8%, 1.2%, and 4.4%. Co-payment incentives to use mail-service pharmacies instead of community pharmacies were associated with higher mail-service utilization rates and with higher costs to plan sponsors. Absence of a co-payment incentive to use mail-service pharmacies was associated with lower mail-service utilization rates and with lower costs to plan sponsors.


People are not passive acceptors of threats to health: endogeneity and its consequences.
The effect of behaviour on health is a major area of contemporary epidemiological enquiry. Most epidemiological studies of the effect of behaviour on health assume that the levels of the behaviour-related variables are determined by factors other than those under study. However, in many instances, obvious examples are breastfeeding and smoking, not only do behaviours affect health but, conversely, individuals take into account their (observable and non-observable) health conditions when making behavioural decisions. In models which allow for the joint determination of health and behaviour, both health and behavioural variables are 'endogenous', that is, determined by forces acting within the model. Through some simple didactic examples it is shown that estimates of the effect of behaviour on health are biased if endogeneity is ignored. Review of the small empirical literature on this subject shows perverse results, such as a negative relationship between the use of prenatal care and infant mortality, when endogeneity is ignored. Standard procedures for taking account of the effects of endogeneity are described briefly.


p16(INK4a) expression and breast cancer risk in women with atypical hyperplasia.
p16, a nuclear protein encoded by the p16(INK4a) gene, is a regulator of cell-cycle regulation. Previous studies have shown that expression of p16 in tissue biopsies of patients with ductal carcinoma in situ (DCIS) is associated with increased risk of breast cancer, particularly when considered in combination with other markers such as Ki-67 and COX-2. Here, we evaluated how expression of p16 in breast tissue biopsies of women with atypical hyperplasia (AH), a putative precursor lesion to DCIS, is associated with subsequent development of cancer. p16 expression was assessed by immunohistochemistry in archival sections from 233 women with AH diagnosed at the Mayo Clinic. p16 expression in the atypical lesions was scored by percentage of positive cells and intensity of staining. We also studied coexpression of p16, with Ki-67 and COX-2, biomarkers of progression in AH. Risk factor and follow-up data were obtained via study questionnaire and medical records. Forty-seven patients (20%) developed breast cancer with a median follow-up of 14.5 years. Staining of p16 was increased in older patients relative to younger patients (P = 0.0025). Although risk of developing breast cancer was not associated with increased p16 expression, joint overexpression of Ki-67 and COX-2 was found to convey stronger risk of breast cancer in the first 10 years after diagnosis as compared with one negative marker (P < 0.01). However, the addition of p16 levels did not strengthen this association. p16 overexpression, either alone or in combination with COX-2 and Ki-67, does not significantly stratify breast cancer risk in women with AH.


On the feasibility of concurrent human TMS-EEG-fMRI measurements.
Simultaneously combining the complementary assets of EEG, functional MRI (fMRI), and transcranial magnetic stimulation (TMS) within one experimental session provides synergetic results, offering insights into brain function that go beyond the scope of each method when used in isolation. The steady increase of concurrent EEG-fMRI, TMS-EEG, and TMS-fMRI studies further underlines the added value of such multimodal imaging approaches. Whereas concurrent EEG-fMRI enables monitoring of brain-wide network dynamics with high temporal and spatial resolution, the combination with TMS provides insights in causal interactions within these networks. Thus the simultaneous use of all three methods would allow studying fast, spatially accurate, and distributed causal interactions in the perturbed system and its functional relevance for intact behavior. Concurrent EEG-fMRI, TMS-EEG, and TMS-fMRI experiments are already technically challenging, and the three-way combination of TMS-EEG-fMRI might yield additional difficulties in terms of hardware strain or signal quality. The present study explored the feasibility of concurrent TMS-EEG-fMRI studies by performing safety and quality assurance tests based on phantom and human data combining existing commercially available hardware. Results revealed that combined TMS-EEG-fMRI measurements were technically feasible, safe in terms of induced temperature changes, allowed functional MRI acquisition with comparable image quality as during concurrent EEG-fMRI or TMS-fMRI, and provided artifact-free EEG before and from 300 ms after TMS pulse application. Based on these empirical findings, we discuss the conceptual benefits of this novel complementary approach to investigate the working human brain and list a number of precautions and caveats to be heeded when setting up such multimodal imaging facilities with current hardware.


Beliefs about sleep in disorders characterized by sleep and mood disturbance.
Maladaptive sleep beliefs play an important role in primary insomnia, but their role in other disorders with concomitant sleep disruption has rarely been explored. Thus, this study investigated the link between insomnia and sleep beliefs in five groups (N=422): primary insomnia (PI), good sleepers (GS), fibromyalgia (FM), major depressive disorder (MDD), and Community Sleep Clinic patients with comorbid insomnia and mood disturbance (CSC). Groups were compared on the Dysfunctional Beliefs and Attitudes about Sleep (DBAS-16) scale. Unlike the GS group, the MDD, FM, and CSC groups had elevated DBAS-16 scores that were similar to, or more pathological than, those of primary insomnia sufferers. Only some of the differences were removed after controlling for depression. Like primary insomnia patients, other sleep-disturbed patient groups have problematic sleep beliefs. Depression was not sufficient to account for all elevations in beliefs. The presence of maladaptive sleep beliefs in these patients suggests that belief-targeted treatment might be helpful in alleviating sleep complaints.


Polymerase chain reaction, nuclease digestion, and mass spectrometry based assay for the trinucleotide repeat status of the fragile X mental retardation 1 gene.
CGG repeat expansions in the 5' noncoding region of the fragile X mental retardation 1 gene (FMR1) give rise to both neurodevelopmental and neurodegenerative human diseases depending on the length of the expansion. Expansions beyond 200 repeats (full mutation) generally result in gene silencing and fragile X syndrome (FXS), the leading heritable form of cognitive impairment and autism. Smaller expansions (55-200 CGG repeats; "premutation") give rise to the neurodegenerative disorder fragile X-associated tremor/ataxia syndrome (FXTAS) through an entirely distinct, toxic mRNA gain-of-function mechanism. A rapid means for both high-risk and newborn screening for allele size would provide a greater opportunity for early intervention and family counseling as well as furnish critical data on repeat size distribution and expanded allele frequencies. In the current work, we propose a novel mass spectrometry (MS) based method for the rapid identification of expanded CGG repeats to complement a recently described polymerase chain reaction (PCR) method for large population screening. In this combined approach, the optimized PCR method is used to amplify the relevant region of FMR1, followed by extensive nonspecific nuclease digestion. The resulting oligonucleotides are analyzed by MS in a manner that provides the relative proportion of triplet repeat oligonucleotides in seconds per sample. This assay enables swift and reproducible detection of expanded CGG alleles using a single blood spot and in principle is suitable for large scale studies and newborn screening. Moreover, this analytical scheme establishes a unique new intersection of MS with molecular biology, with potential for significant interdisciplinary impact.


Restored Self: A Phenomenological Study of Pain Relief by Cannabis.
To explore the subjective experience of pain relief by cannabis. Qualitative data were collected through in-depth semistructured interviews. Interview transcripts were analyzed using interpretative phenomenological analysis (IPA). Nineteen patients, aged 28 to 79, who were treated with medical cannabis under the supervision of a pain clinic in Israel. Three key themes that emerged from the analysis were explored: 1) the Sigh of Relief, describing the corporal sensation of using cannabis, including a sense of relaxation and reduction in pain; 2) the Return to Normality, describing the comprehensive effect of using cannabis, including an increased ability to sleep, focus, and function; and 3) the Side Effects of using cannabis. We propose the term Restored Self to conceptualize the effect of medical cannabis. Restored Self is the experience of regaining one's sense of self, sense of normality, and sense of control over one's life.


Neurophysiological Correlates and Differential Drug Response in Subjects With a Family History of an Alcohol Use Disorder.
A family history of an alcohol use disorder (AUD) has been shown to increase one's risk of developing an AUD. Additionally, a positive family history of AUD (family history positive (FHP)) has neurobiological and neuropsychopharmacological consequences, and this review summarizes differential drug response as well as neuroanatomical and neurocognitive correlates. FHP status is related to altered responses to a number of drugs, including substances with abuse liability like alcohol, opioids, amphetamines, and ketamine. FHP individuals demonstrate fewer aversive effects and more rewarding response to both alcohol and subanesthetic dose ketamine. Ketamine is a rapid-acting antidepressant, and several studies have reported that ketamine is more effective for FHP treatment-resistant depressed individuals. In short, the reviewed neurophysiological differences may contribute to ketamine's enhanced antidepressant efficacy in FHP patients. Volumetric differences in the amygdala, nucleus accumbens, neocortex, and cerebellum are commonly reported. Furthermore, FHP has also been associated with altered neurocognitive performance, e.g., increased impulsivity. The imaging and psychological literature supports a neurodevelopmental lag hypothesis in FHP youth. The review will further discuss these findings in depth.


Body Mass Index Categories and Attained Height in Portuguese Adults.
To analyze the associations between height and BMI categories in a Portuguese representative sample. This is a cross-sectional study with a representative sample of 32,644 Portuguese adults (52.4% females). Sociodemographic and lifestyle characteristics were obtained along with self-reported height and weight. We performed generalized linear models to assess the differences in attained height across BMI categories; analyses were adjusted for age, gender, education, family income per month, proxy reporting information, dietary patterns, and smoking. BMI categories included underweight and normal weight (46.4%), overweight (37.6%), obese class I and II (15.2%), and obese class III (0.8%). Adults with normal weight had a significantly higher height (females +7 cm and males +5 cm) when compared to obese class III. As BMI categories increased, height decreased. In females and males, after adjusting for confounders, estimates of attained height decreased when compared to the unadjusted model (β = -0.049, 95% CI = -0.050; -0.049 and β = -0.030, 95% CI = -0.031; -0.029, respectively), although they remained still significant. Our results suggest a significant difference in attained height between BMI categories. Future intervention programs aiming at preventing overweight and obesity should monitor sociodemographic, health and environmental conditions that affect attained height potential.


A note on a discussion group study of public preferences regarding priorities in the allocation of donor kidneys.
To explore whether and how people wish to give differential priority based on certain characteristics of the potential recipient of a donor kidney. A random sample of people resident in York was invited to attend two focus group meetings each, a fortnight apart. The City of York. Twenty-three randomly chosen people meeting in four groups of five or six. Those factors that people think should be taken into account when allocating donor kidneys, in addition to the expected benefits from transplantation. People are willing and able to distinguish between potential recipients of a kidney transplantation according to a range of characteristics beyond the expected benefits from treatment. There is a clear consensus across the four groups that one of the most important considerations is what will happen to the patient without treatment, and so priority is given to those with a poor prognosis. There is also a strong view that priority should be given to younger patients and to those with dependants. The time spent waiting for a transplant is also important, but less so. A sample of the general public, after discussion and debate, wish to take account of a number of patient characteristics when allocating donor kidneys. There is some degree of consensus about what these factors should be and this suggests that it might be possible to develop a set of guidelines for the allocation of donor kidneys.


Impairment of protein homeostasis and decline of proteasome activity in microglial cells from adult Wistar rats.
Common symptoms of different neurodegenerative diseases start to develop in the second half of the human life. Several of these diseases, including Alzheimer's and Parkinson's disease, are accompanied by severe disturbances of protein metabolism and homeostasis in the brain. Because microglial cells are, to some extent, responsible for the maintenance of this homeostasis, age-related functional changes of the microglia are important. We established, therefore, the preparation of cultures of primary microglial cells isolated from adult animals in comparison to the widely used standard model, primary microglial cells isolated from newborn animals. In addition, we investigated changes in the activation and in the protein homeostasis within these cells. The protein turnover seems to be significantly impaired in microglial cells isolated from adult animals and this seems to be accompanied by a decline in proteasomal function, but not in the protease content. We were also able to demonstrate higher cell surface molecule expression and a higher basal NO release of microglia isolated from adult animals in comparison to the microglia isolated from newborn rats; however, the PMA stimulated oxidative burst was abolished completely in cells from adult animals. Microglia from adult animals were also not able to upregulate their protein metabolism after activation. From these investigations it was concluded that microglial cells from adult animals have significantly different metabolic properties in comparison to the widely used microglial cells from newborn animals.


Like will to like: abundances of closely related species can predict susceptibility to intestinal colonization by pathogenic and commensal bacteria.
The intestinal ecosystem is formed by a complex, yet highly characteristic microbial community. The parameters defining whether this community permits invasion of a new bacterial species are unclear. In particular, inhibition of enteropathogen infection by the gut microbiota ( = colonization resistance) is poorly understood. To analyze the mechanisms of microbiota-mediated protection from Salmonella enterica induced enterocolitis, we used a mouse infection model and large scale high-throughput pyrosequencing. In contrast to conventional mice (CON), mice with a gut microbiota of low complexity (LCM) were highly susceptible to S. enterica induced colonization and enterocolitis. Colonization resistance was partially restored in LCM-animals by co-housing with conventional mice for 21 days (LCM(con21)). 16S rRNA sequence analysis comparing LCM, LCM(con21) and CON gut microbiota revealed that gut microbiota complexity increased upon conventionalization and correlated with increased resistance to S. enterica infection. Comparative microbiota analysis of mice with varying degrees of colonization resistance allowed us to identify intestinal ecosystem characteristics associated with susceptibility to S. enterica infection. Moreover, this system enabled us to gain further insights into the general principles of gut ecosystem invasion by non-pathogenic, commensal bacteria. Mice harboring high commensal E. coli densities were more susceptible to S. enterica induced gut inflammation. Similarly, mice with high titers of Lactobacilli were more efficiently colonized by a commensal Lactobacillus reuteri(RR) strain after oral inoculation. Upon examination of 16S rRNA sequence data from 9 CON mice we found that closely related phylotypes generally display significantly correlated abundances (co-occurrence), more so than distantly related phylotypes. Thus, in essence, the presence of closely related species can increase the chance of invasion of newly incoming species into the gut ecosystem. We provide evidence that this principle might be of general validity for invasion of bacteria in preformed gut ecosystems. This might be of relevance for human enteropathogen infections as well as therapeutic use of probiotic commensal bacteria.


Fatal acute pancreatitis.
Review of all deaths from acute pancreatitis recorded at Glasgow Royal Infirmary between 1974 and 1984 identified 126 patients, 53 (42%) of whom had pancreatitis first diagnosed at necropsy. Aetiologies of the fatal attacks of pancreatitis included gall stones (30%), alcohol (15%), other identified aetiological factors (17%), and was unknown (38%). Overall mortality fell from 14.9% in the early half of the study to 10.8% in the latter half although in the 73 patients in whom the diagnosis of acute pancreatitis was made during life, the mortality rate was unchanged throughout. Within the group of 73 patients diagnosed during life deaths from gall stone pancreatitis have fallen by almost 50% suggesting that improved treatment of this subgroup may have occurred. The findings of this study lend support to the concept of early, complete clearance of calculi from the biliary tree, either by an endoscopic or surgical approach.


The Tayside infant morbidity and mortality study: effect on health of using gas for cooking.
The relation between respiratory illness and the use of gas for cooking was examined from data on 1565 infants born to mothers who were primigravidas living in Dundee in 1980. Episodes of, and admissions to hospital for, respiratory illness were recorded during the first year of life. Both admissions and episodes were more common in infants from families using gas for cooking or heating than in infants from families using any other type of cooking or heating, but the differences were not significant. Results from this and other studies show that there is probably a small relation between respiratory illness and the use of gas appliances without a flue. To show convincingly whether such a relation exists might require a survey of 18 000-23 000 subjects. Respiratory illness was, however, strongly and positively related to parental smoking, a finding that is often made even in small studies.


Transient and persistent hypercalcaemia in patients treated by maintenance haemodialysis.
In a group of 32 patients with terminal renal failure the initial hypocalcaemia was corrected after two months' adequate maintenance haemodialysis. In seven patients hypercalcaemia occurred with a peak incidence after about six months' treatment. In six of these patients hypercalcaemia was transient and the plasma calcium became normal with haemodialysis alone. In one patient the hypercalcaemia was persistent and the plasma calcium reverted to normal only after subtotal parathyroidectomy. This patient had no radiological bone disease, a normal alkaline phosphatase, and no metastatic calcification of the soft tissues.It is concluded that in some patients with terminal renal failure treated with maintenance haemodialysis autonomy of the parathyroids becomes evident in the absence of bone disease or a raised plasma alkaline phosphatase, and that subsequently with continued dialysis there is a spontaneous involution towards normal parathyroid function.


Pore-level mechanics of foam generation and coalescence in the presence of oil.
The stability of foam in porous media is extremely important for realizing the advantages of foamed gas on gas mobility reduction. Foam texture (i.e., bubbles per volume of gas) achieved is dictated by foam generation and coalescence processes occurring at the pore-level. For foam injection to be widely applied during gas injection projects, we need to understand these pore-scale events that lead to foam stability/instability so that they are modeled accurately. Foam flow has been studied for decades, but most efforts focused on studying foam generation and coalescence in the absence of oil. Here, the extensive existing literature is reviewed and analyzed to identify open questions. Then, we use etched-silicon micromodels to observe foam generation and coalescence processes at the pore-level. Special emphasis is placed on foam coalescence in the presence of oil. For the first time, lamella pinch-off as described by Myers and Radke [40] is observed in porous media and documented. Additionally, a new mechanism coined "hindered generation" is found. Hindered generation refers to the role oil plays in preventing the successful formation of a lamella following snap-off near a pore throat.


Climate change and respiratory disease: European Respiratory Society position statement.
Climate change will affect individuals with pre-existing respiratory disease, but the extent of the effect remains unclear. The present position statement was developed on behalf of the European Respiratory Society in order to identify areas of concern arising from climate change for individuals with respiratory disease, healthcare workers in the respiratory sector and policy makers. The statement was developed following a 2-day workshop held in Leuven (Belgium) in March 2008. Key areas of concern for the respiratory community arising from climate change are discussed and recommendations made to address gaps in knowledge. The most important recommendation was the development of more accurate predictive models for predicting the impact of climate change on respiratory health. Respiratory healthcare workers also have an advocatory role in persuading governments and the European Union to maintain awareness and appropriate actions with respect to climate change, and these areas are also discussed in the position statement.