Efficient Volume Exploration Using the Gaussian Mixture Model.
The multidimensional transfer function is a flexible and effective tool for exploring volume data. However, designing an appropriate transfer function is a trial-and-error process and remains a challenge. In this paper, we propose a novel volume exploration scheme that explores volumetric structures in the feature space by modeling the space using the Gaussian mixture model (GMM). Our new approach has three distinctive advantages. First, an initial feature separation can be automatically achieved through GMM estimation. Second, the calculated Gaussians can be directly mapped to a set of elliptical transfer functions (ETFs), facilitating a fast pre-integrated volume rendering process. Third, an inexperienced user can flexibly manipulate the ETFs with the assistance of a suite of simple widgets, and discover potential features with several interactions. We further extend the GMM-based exploration scheme to time-varying data sets using an incremental GMM estimation algorithm. The algorithm estimates the GMM for one time step by using itself and the GMM generated from its previous steps. Sequentially applying the incremental algorithm to all time steps in a selected time interval yields a preliminary classification for each time step. In addition, the computed ETFs can be freely adjusted. The adjustments are then automatically propagated to other time steps. In this way, coherent user-guided exploration of a given time interval is achieved. Our GPU implementation demonstrates interactive performance and good scalability. The effectiveness of our approach is verified on several data sets.


Longitudinal gradients of KCNQ4 expression in spiral ganglion and cochlear hair cells correlate with progressive hearing loss in DFNA2.
Mutations in the human KCNQ4 gene were recently found by Kubisch et al. [Cell 96 (1999) 437-446] to cause a non-syndromic, autosomal dominant, progressive hearing loss, DFNA2. The mouse Kcnq4 orthologue was previously localized to the outer hair cells (OHCs) of the inner ear, suggesting the pathophysiological effects were due to dysfunctional OHCs. Yet, OHC dysfunction does not provide a plausible explanation for the progressive nature of the frequency specific hearing loss. We have re-examined and extended the expression analyses of KCNQ4 in the murine inner ear using RT-PCR and whole mount in situ hybridization. Our results confirmed that the rat KCNQ4 orthologue is expressed in both inner and outer hair cells. Reciprocal longitudinal gradients were found in inner hair cells (IHCs) and OHCs. The strongest expression of KCNQ4 in IHCc was in the base of the cochlea and in the apex for OHCs. Similar to the IHCs, a basal to apical gradient was present in the spiral sensory neurons. IHCs mediate hearing via their afferent sensory neurons, whereas OHCs function as active cochlear amplifiers. The complete absence of OHCs leads only to severe sensitivity reduction, but not complete hearing loss. Our data suggest that the primary defect leading to initial high frequency loss and subsequent progressive hearing loss for all frequencies may be due to spiral ganglion and/or IHC dysfunction, rather than an OHC aberration.


The association between acculturation patterns and mental health symptoms among Eritrean and Sudanese asylum seekers in Israel.
Past research has documented the role acculturation plays in the process of adjustment to new cultures among migrants. Yet little attention has been paid thus far to the role of acculturation in the context of forced migration. In this study we examined the association between acculturation patterns and mental health symptoms among a convenience sample of Eritrean and Sudanese asylum seekers (n = 118) who accessed health services at the Physicians for Human Rights Open-Clinic in Israel. Participants completed measures on sociodemographic information as well as detention history, mental health symptoms, exposure to traumatic events, and acculturation pattern, in their native language upon accessing services. Consistent with our predictions, findings showed that acculturation predicted depressive symptoms among asylum seekers beyond the effect of history of detention and reports of experiences of traumatic events. Assimilated compared with integrated asylum seekers reported higher depressive symptoms. Findings draw attention to the paradox of assimilation, and the mental health risks it poses among those wishing to integrate into the new culture at the expanse of their original culture. Asylum seekers may be particularly vulnerable to the risks of assimilation in the restrictive policies that characterize many industrial countries in recent years.


Changes in Prefrontal Cortical Activity During Walking and Cognitive Functions Among Patients With Parkinson's Disease.
Background: Walking becomes more and more degraded as Parkinson's Disease (PD) progresses. Previous research examined factors contributing to this deterioration. Among them, changes in brain cortical activity during walking have been less studied in this clinical population. Objectives: This study aimed to: (1) investigate changes in dorsolateral prefrontal cortex (DLPFC) activation during usual walking and dual-task walking conditions in patients with PD; (2) examine the association between cortical activity and behavioral/cognitive outcomes; and (3) explore which factors best predict increased activation of the DLPFC during usual walking. Methods: Eighteen patients with early stage PD and 18 controls performed 4 conditions: (1) standing while subtracting, (2) usual walking, (3) walking while counting forward, and (4) walking while subtracting. Cortical activity in DLPFC, assessed by changes in oxy-hemoglobin (ΔHbO2) and deoxy-hemoglobin (ΔHbR), was measured using functional near infrared spectroscopy (fNIRS). Gait performance was recorded using wearables sensors. Cognition was also assessed using neuropsychological tests, including the Trail Making Test (TMT). Results: DLPFC activity was higher in patients compared to controls during both usual walking and walking while subtracting conditions. Patients had impaired walking performance compared to controls only during walking while subtracting task. Moderate-to-strong correlations between ΔHbO2 and coefficients of variation of all gait parameters were found for usual walking and during walking while counting forward conditions. Part-B of TMT predicted 21% of the variance of ΔHbO2 during usual walking after adjustment for group status. Conclusions: The increased DLPFC activity in patients during usual walking suggests a potential compensation for executive deficits. Understanding changes in DLPFC activity during walking may have implications for rehabilitation of gait in patients with PD.


A genetic linkage map for the ectomycorrhizal fungus Laccaria bicolor and its alignment to the whole-genome sequence assemblies.
A genetic linkage map for the ectomycorrhizal basidiomycete Laccaria bicolor was constructed from 45 sib-homokaryotic haploid mycelial lines derived from the parental S238N strain progeny. For map construction, 294 simple sequence repeats (SSRs), single-nucleotide polymorphisms (SNPs), amplified fragment length polymorphisms (AFLPs) and random amplified polymorphic DNA (RAPD) markers were employed to identify and assay loci that segregated in backcross configuration. Using SNP, RAPD and SSR sequences, the L. bicolor whole-genome sequence (WGS) assemblies were aligned onto the linkage groups. A total of 37.36 Mbp of the assembled sequences was aligned to 13 linkage groups. Most mapped genetic markers used in alignment were colinear with the sequence assemblies, indicating that both the genetic map and sequence assemblies achieved high fidelity. The resulting matrix of recombination rates between all pairs of loci was used to construct an integrated linkage map using JoinMap. The final map consisted of 13 linkage groups spanning 812 centiMorgans (cM) at an average distance of 2.76 cM between markers (range 1.9-17 cM). The WGS and the present linkage map represent an initial step towards the identification and cloning of quantitative trait loci associated with development and functioning of the ectomycorrhizal symbiosis.


Comparing efficacy and tolerability of ibuprofen and paracetamol in fever.
The purpose of this study was to compare antipyretic activity and evaluate tolerability of ibuprofen and paracetamol suspension in the treatment of febrile children. It was designed as a double blind, parallel group, multiple dose study comparing ibuprofen (20 mg/kg/24 hours) with paracetamol (50 mg/kg/24 hours) given at six hourly intervals for a maximum of 12 doses. Children on paediatric wards between the ages of 0.2 and 12 years, with fever as defined by an axillary temperature > or = 37.5 degrees C, were included. The main outcome measures were: change in axillary temperature; palatability of medication; changes in irritability and clinical condition; overall efficacy at the end of treatment; and number and nature of adverse events. The mean temperature change from baseline at four hours was -1.8 degrees C and -1.6 degrees C in ibuprofen and paracetamol groups respectively. In both groups: median palatability score was 'no reaction'; median irritability score at end point was 'not irritable'; median score for change in clinical condition was 'improved'; and median score for overall efficacy was 'good effect'. The proportion of patients experiencing adverse events was similar in both groups, the majority of events having doubtful or no relationship to therapy and being mild in severity. In conclusion, ibuprofen suspension was as effective and well tolerated as paracetamol in treatment of fever in young children.


Bacterial and host factors implicated in nasal carriage of methicillin-resistant Staphylococcus aureus in mice.
Nasal carriage is a major risk factor for Staphylococcus aureus infection, especially for methicillin-resistant strains (MRSA). Using a mouse model of nasal carriage, we have compared several S. aureus strains and demonstrated increased colonization levels by MRSA in cystic fibrosis transmembrane conductance regulator-deficient mice and Toll-like receptor 2 (TLR2)-deficient mice but not TLR4-deficient mice.


Single-cell membrane drug delivery using porous pen nanodeposition.
Delivering molecules onto the plasma membrane of single cells is still a challenging task in profiling cell signaling pathways with single cell resolution. We demonstrated that a large quantity of molecules could be targeted and released onto the membrane of individual cells to trigger signaling responses. This is achieved by a porous pen nanodeposition (PPN) method, in which a multilayer porous structure, serving as a reservoir for a large amount of molecules, is formed on an atomic force microscope (AFM) tip using layer-by-layer assembly and post processing. To demonstrate its capability for single cell membrane drug delivery, PPN was employed to induce a calcium flux triggered by the binding of released antibodies to membrane antigens in an autoimmune skin disease model. This calcium signal propagates from the target cell to its neighbors in a matter of seconds, proving the theory of intercellular communication through cell-cell junctions. Collectively, these results demonstrated the effectiveness of PPN in membrane drug delivery for single cells; to the best of our knowledge, this is the first technique that can perform the targeted transport and delivery in single cell resolution, paving the way for probing complex signaling interactions in multicellular settings.


Combined Interactions of Plant Homeodomain and Chromodomain Regulate NuA4 Activity at DNA Double-Strand Breaks.
DNA double-strand breaks (DSBs) represent one of the most threatening lesions to the integrity of genomes. In yeast Saccharomyces cerevisiae, NuA4, a histone acetylation complex, is recruited to DSBs, wherein it acetylates histones H2A and H4, presumably relaxing the chromatin and allowing access to repair proteins. Two subunits of NuA4, Yng2 and Eaf3, can interact in vitro with methylated H3K4 and H3K36 via their plant homeodomain (PHD) and chromodomain. However, the roles of the two domains and how they interact in a combinatorial fashion are still poorly characterized. In this study, we generated mutations in the PHD and chromodomain that disrupt their interaction with methylated H3K4 and H3K36. We demonstrate that the combined mutations in both the PHD and chromodomain impair the NuA4 recruitment, reduce H4K12 acetylation at the DSB site, and confer sensitivity to bleomycin that induces DSBs. In addition, the double mutant cells are defective in DSB repair as judged by Southern blot and exhibit prolonged activation of phospho-S129 of H2A. Cells harboring the H3K4R, H3K4R, K36R, or set1Δ set2Δ mutant that disrupts H3K4 and H3K36 methylation also show very similar phenotypes to the PHD and chromodomain double mutant. Our results suggest that multivalent interactions between the PHD, chromodomain, and methylated H3K4 and H3K36 act in a combinatorial manner to recruit NuA4 and regulate the NuA4 activity at the DSB site.


Depiction of intracranial vessels with MRA: utility of magnetization transfer saturation and gadolinium.
The purpose of this work was to quantitate the individual and combined effects of magnetization transfer (MT) saturation and gadolinium (Gd) on the visualization of intracranial vessels with MR angiography (MRA). Thirty-five subjects underwent two three-dimensional time-of-flight MRA sequences without and with MT and/or Gd. There were 14 MR angiograms without Gd or MT, 18 with MT only, 17 with Gd only, and 21 with both Gd and MT. On a projection image, a region of interest was drawn to delineate the arteries in the middle cerebral artery territory. The total area of blood vessels in the region of interest was calculated for each MR angiogram. Mean vessel areas for the four types of MRA were compared with analysis of variance. MRA with either MT or Gd alone showed significantly more vessel area than MRA without either (p < 0.05). MRA with MT alone and MRA with Gd alone were not different from each other (p = 0.29). The improvement in vessel area measured by using MT and Gd together was significantly more than expected from the cumulative improvement of adding each alone (p < 0.05). Combining MT and Gd synergistically improved the visualization of intracranial vessels on MRA.


Modeling Commercial Freshwater Turtle Production on US Farms for Pet and Meat Markets.
Freshwater turtles are being exploited for meat, eggs, traditional medicine, and pet trade. As a response, turtle farming became a booming aquaculture industry in the past two decades, specifically in the southeastern states of the United States of America (US) and across Southeast Asia. However, US turtle farms are currently producing turtles only for the pet trade while commercial trappers remain focused on catching the largest individuals from the wild. In our analyses we have created a biological and economic model that describes farming operations on a representative turtle farm in Louisiana. We first modeled current production of hatchling and yearling red-eared slider turtles (Trachemys scripta elegans) (i.e., traditional farming) for foreign and domestic pet markets, respectively. We tested the possibility of harvesting adult turtles from the breeding stock for sale to meat markets to enable alternative markets for the farmers, while decreasing the continued pressures on wild populations (i.e., non-traditional farming). Our economic model required current profit requirements of ~$13/turtle or ~$20.31/kg of meat from non-traditional farming in order to acquire the same profit as traditional farming, a value which currently exceeds market values of red-eared sliders. However, increasing competition with Asian turtle farms and decreasing hatchling prices may force the shift in the US toward producing turtles for meat markets. In addition, our model can be modified and applied to more desirable species on the meat market once more knowledge is acquired about species life histories and space requirements under farmed conditions.


Retrospective case series analysis of penicillin allergy testing in a UK specialist regional allergy clinic.
Penicillin allergy is the most common drug allergy. Skin testing for the major (PPL) and minor determinants (MDMs) of penicillin offers increased sensitivity and specificity over in vitro testing alone. Following a worldwide absence of reagents, a new kit was licensed in the UK in 2008 (Diater, Spain) and this report evaluates its use in a UK specialist allergy clinic. Prospective data on 50 consecutive patients tested with the new reagents were collected. The departmental protocol is adapted from the 2003 EAACI position paper. 14% (7/50) and 12% (6/50) of patients were diagnosed with immediate and non-immediate reactions respectively. The negative predictive value of the PPL and MDM reagents at the neat concentration for an immediate reaction was 93% (true negatives 37, false negatives 3). Two patients experienced systemic reactions to DPT in the absence of demonstrable specific IgE. None of the patients were diagnosed using skin prick testing alone or at lower concentrations of IDT. Five patients were diagnosed at the IDT stage and two at the DPT stage in the absence of demonstrable specific IgE. Six patients were diagnosed with non-immediate reactions, two on IDT alone and four following IDT and DPT. The new PPL and MDM determinants offer enhanced sensitivity when evaluating β-lactam hypersensitivity; however, there are limitations to the current testing regimens. The UK would benefit from local guidelines, which incorporate the new reagents and acknowledge the high amoxicillin prescription rate and the relatively lower specialist-to-patient ratio in this country.


Choroid plexus cyst and chordoid glioma. Report of two cases.
Several types of mass lesions may occur in the third and lateral ventricles. Typically they arise from the lining of the ventricular cavity or from contiguous structures, by extension into the ventricle. The authors describe two patients, each of whom presented with a different rare lesion of the ventricular system. The first was a 53-year-old woman with a history of hypertension who sustained a blunt traumatic injury to the occipital region and subsequently developed a progressively worsening right-sided headache. Radiological examinations over the next 2 years revealed an enlarged right lateral ventricle and, ultimately, a choroid plexus cyst in its anterior and middle third, near the foramen of Monro, which is a rare location for these lesions. The cyst was removed en bloc, and follow-up examinations showed a significant improvement in her headache and a minimal differences in size between right and left ventricles. The authors also describe a 57-year-old man with hypertension, diabetes mellitus, and an old mycardial infarct, who presented to an outside institution with a progressively worsening headache, generalized malaise, and loss of olfactory sensation. Diagnostic imaging revealed a 1.5-cm oval lesion centered in the lamina terminalis region, an open craniotomy was performed, and evaluation of a biopsy sample demonstrated the mass to be a chordoid glioma of the third ventricle, a recently described glioma subtype. Two days after surgery, he suffered a left parietal stroke and an anterior mycardial infarction. After convalescing, he presented to The University of Texas M. D. Anderson Cancer Center for radiotherapy and follow up; 7 months later he was readmitted complaining of headache, short-term memory loss, and worsening confusion and disorientation. Neuroimaging revealed progression of the tumor (now 2 cm in diameter), which was removed by gross-total resection. His headache resolved immediately, and 2 months later his only complaint was of episodes of confusion. Three weeks later he died of a massive myocardial infarction. These two patients represent the sixth case of an adult with a choroid plexus cyst in the anterior lateral ventricle and the 19th case of an adult with a chordoid glioma of the third ventricle, respectively.


High cancer mortality for US-born Latinos: evidence from California and Texas.
Latinos born in the US, 36 million, comprise 65% of all US Latinos. Yet their cancer experience is nearly always analyzed together with their foreign-born counterparts, 19 million, who constitute a steady influx of truly lower-risk populations from abroad. To highlight specific cancer vulnerabilities for US-born Latinos, we compare their cancer mortality to the majority non-Latino white (NLW) population, foreign-born Latinos, and non-Latino blacks. We analyzed 465,751 cancer deaths from 2008 to 2012 occurring among residents of California and Texas, the two most populous states, accounting for 47% of US Latinos. This cross-sectional analysis, based on granular data obtained from death certificates on cause of death, age, race, ethnicity and birthplace, makes use of normal standardization techniques and negative binomial regression models. While Latinos overall have lower all-cancers-combined mortality rates than NLWs, these numbers were largely driven by low rates among the foreign born while mortality rates for US-born Latinos approach those of NLWs. Among Texas males, rates were 210 per 100,000 for NLWs and 166 for Latinos combined, but 201 per 100,000 for US-born Latinos and 125 for foreign-born Latinos. Compared to NLWs, US-born Latino males in California had mortality rate ratios of 2.83 (95% CI: 2.52-3.18) for liver cancer, 1.44 (95% CI: 1.30-1.61) for kidney cancer, and 1.25 (95% CI: 1.17-1.34) for colorectal cancer (CRC). Texas results showed a similar site-specific pattern. Specific cancer patterns for US-born Latinos, who have relatively high cancer mortality, similar overall to NLWs, are masked by aggregation of all Latinos, US-born and foreign-born. While NLWs had high mortality for lung cancer, US-born Latinos had high mortality for liver, kidney and male colorectal cancers. HCV testing and reinforcement of the need for CRC screening should be a priority in this specific and understudied population. The unprecedented proximity of overall rates between NLWs and US-born Latino populations runs counter to the prevailing narrative of Latinos having significantly lower cancer risk and mortality. Birthplace data are critical in detecting meaningful differences among Latinos; these findings merit not only clinical but also public health attention.


Identifying Factors Underlying the Decision for Sickle Cell Carrier Screening Among African Americans Within Middle Reproductive Age.
Guidelines recommend that African Americans know their sickle cell trait status to inform reproductive health decisions. Few studies have applied a behavioral theory to identify factors associated with sickle cell trait screening to inform intervention targets to increase this behavior. We applied a Sickle Cell Trait Screening Framework to identify factors associated with African Americans' intention to ask for sickle cell trait screening. Participants (N = 300), ages 18 to 35, completed a cross-sectional survey. A three-step sequential ordinary least squares regression analysis identified factors influencing intention. Results indicated socio-demographic factors (age, education), knowledge and fear beliefs (screening knowledge, perceived threat), and reasoned action approach (RAA) constructs were associated with intention. RAA constructs influenced intention over knowledge and fear beliefs with an increase in R2 of .468. Perceived behavioral control was more predictive of intention (β = .576, p < .001). Attitude and perceived norm also had significant weights (β = .325 and β = .192, both p < .001, respectively). Findings from this study can inform strategies (e.g., eliminating costs associated with screening, reducing fear of painful tests) to increase sickle cell trait screening among African Americans. Ultimately, more sickle cell carriers will become aware of their trait status and be able to make informed reproductive health decisions.


Methylglyoxal displays colorectal cancer-promoting properties in the murine models of azoxymethane and CT26 isografts.
Methylglyoxal (MG), a highly reactive carbonyl species (RCS) with pro-oxidant and proinflammatory properties, may be a colon tumor-promoting factor in food and biological systems. In the present study, we found that consumption of MG significantly deteriorated azoxymethane (AOM)-induced colonic preneoplastic lesions in ICR mice, in which biomarkers of oxidative stress and inflammation within the body and feces induced by MG-fueled carbonyl stress may have played important roles. Interestingly, exposure to MG also led to increases in the serum low-density lipoprotein (LDL)/high-density lipoprotein (HDL) ratio and fecal bile acid levels in mice, which may be critical factors involved in MG-induced colonic lesions. Additionally, MG treatment (50mg/kg body weight (BW); intraperitoneally) promoted tumor growth of CT26 isografts in mice partly by carbonyl stress-evoked protumorigenic responses, including low-grade inflammation and oxidative stress. Furthermore, primary tumor cells isolated from mice with MG-induced CT26 isografts had greater proliferative and migratory activities as well as stem-like properties compared to those isolated from the vehicle controls. Excitingly, enhanced expression or activation of proteins that modulate cell survival, proliferation, or migration/invasion was also observed in those cells. In conclusion, it is conceivable that MG-induced carbonyl stress may be the pivotal promoter involved in colon cancer progression.


Quantitative ultrasound measurements of the tibia and calcaneus in comparison with DXA measurements at various skeletal sites.
The performance of quantitative ultrasound (QUS) measurements of the tibia and calcaneus was studied in 109 elderly people (age range 65-87 years). Broadband ultrasound attenuation (BUA) and speed of sound (SOS) were measured at the calcaneus and SOS was assessed at the tibia. Short-term precision of tibial QUS was studied in 16 volunteers. The coefficient of variation (CV) was 0.4% and the standardized CV (sCV) was 4.4%. We compared the calcaneal and tibial QUS measurements with bone mineral density (BMD) measurements of the lumbar spine, femoral neck, trochanter and total body assessed by dual-energy X-ray absorptiometry (DXA). Calcaneal QUS correlated better with BMD at various skeletal sites than tibial QUS. Calcaneal BUA showed higher correlations with BMD values of the lumbar spine, femoral neck, trochanter and total body than calcaneal and tibial SOS (r = 0.48-0.64, r = 0.30-0.47, r = 0.35-0.47, respectively; p < 0.001). Body weight modified the relationships between calcaneal and tibial QUS and BMD measurements of the hip. Higher body weight was associated with higher BMD values at the femoral neck and trochanter for the same calcaneal and tibial QUS values. After adjustments for body weight correlations of tibial and calcaneal QUS with BMD improved and were very similar. This suggests that correction for body weight is important and could add to the predictive value of QUS measurements.


Immunomodulatory drugs disrupt the cereblon-CD147-MCT1 axis to exert antitumor activity and teratogenicity.
Immunomodulatory drugs (IMiDs), such as thalidomide and its derivatives lenalidomide and pomalidomide, are key treatment modalities for hematologic malignancies, particularly multiple myeloma (MM) and del(5q) myelodysplastic syndrome (MDS). Cereblon (CRBN), a substrate receptor of the CRL4 ubiquitin ligase complex, is the primary target by which IMiDs mediate anticancer and teratogenic effects. Here we identify a ubiquitin-independent physiological chaperone-like function of CRBN that promotes maturation of the basigin (BSG; also known as CD147) and solute carrier family 16 member 1 (SLC16A1; also known as MCT1) proteins. This process allows for the formation and activation of the CD147-MCT1 transmembrane complex, which promotes various biological functions, including angiogenesis, proliferation, invasion and lactate export. We found that IMiDs outcompete CRBN for binding to CD147 and MCT1, leading to destabilization of the CD147-MCT1 complex. Accordingly, IMiD-sensitive MM cells lose CD147 and MCT1 expression after being exposed to IMiDs, whereas IMiD-resistant cells retain their expression. Furthermore, del(5q) MDS cells have elevated CD147 expression, which is attenuated after IMiD treatment. Finally, we show that BSG (CD147) knockdown phenocopies the teratogenic effects of thalidomide exposure in zebrafish. These findings provide a common mechanistic framework to explain both the teratogenic and pleiotropic antitumor effects of IMiDs.


Influence of common genetic variation on blood lipid levels, cardiovascular risk, and coronary events in two British prospective cohort studies.
The aim of this study was to quantify the collective effect of common lipid-associated single nucleotide polymorphisms (SNPs) on blood lipid levels, cardiovascular risk, use of lipid-lowering medication, and risk of coronary heart disease (CHD) events. Analysis was performed in two prospective cohorts: Whitehall II (WHII; N = 5059) and the British Women's Heart and Health Study (BWHHS; N = 3414). For each participant, scores were calculated based on the cumulative effect of multiple genetic variants influencing total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TG). Compared with the bottom quintile, individuals in the top quintile of the LDL-C genetic score distribution had higher LDL-C {mean difference of 0.85 [95% confidence interval, (CI) = 0.76-0.94] and 0.63 [95% CI = 0.50-0.76] mmol/l in WHII and BWHHS, respectively}. They also tended to have greater odds of having 'high-risk' status (Framingham 10-year cardiovascular disease risk >20%) [WHII: odds ratio (OR) = 1.36 (0.93-1.98), BWHHS: OR = 1.49 (1.14-1.94)]; receiving lipid-lowering treatment [WHII: OR = 2.38 (1.57-3.59), BWHHS: OR = 2.24 (1.52-3.29)]; and CHD events [WHII: OR = 1.43 (1.02-2.00), BWHHS: OR = 1.31 (0.99-1.72)]. Similar associations were observed for the TC score in both studies. The TG score was associated with high-risk status and medication use in both studies. Neither HDL nor TG scores were associated with the risk of coronary events. The genetic scores did not improve discrimination over the Framingham risk score. At the population level, common SNPs associated with LDL-C and TC contribute to blood lipid variation, cardiovascular risk, use of lipid-lowering medications and coronary events. However, their effects are too small to discriminate future lipid-lowering medication requirements or coronary events.


Inhibition of dengue virus replication by diisopropyl chrysin-7-yl phosphate.
Dengue fever is a tropical disease and caused by dengue virus (DENV), which is transmitted by mosquitoes and infects about 400 million people annually. With the development of international trade and travel, China is facing a growing threat. Over 40 thousands of people were infected during the 2014 DENV outbreak in Guangdong. Neither licensed vaccine nor therapeutic drug has been available. In this report, we isolated two clinical DENV strains. The full-length genome was sequenced and characterized. We also applied a flavonoid, CPI, into an anti-DENV assay. Replication of viral RNA and expression of viral protein was all strongly inhibited. These results indicated that CPI may serve as potential protective agents in the treatment of patients with chronic DENV infection.


Eating disorder predisposition is associated with ESRRA and HDAC4 mutations.
Anorexia nervosa and bulimia nervosa are common and severe eating disorders (EDs) of unknown etiology. Although genetic factors have been implicated in the psychopathology of EDs, a clear biological pathway has not been delineated. DNA from two large families affected by EDs was collected, and mutations segregating with illness were identified by whole-genome sequencing following linkage mapping or by whole-exome sequencing. In the first family, analysis of twenty members across three generations identified a rare missense mutation in the estrogen-related receptor α (ESRRA) gene that segregated with illness. In the second family, analysis of eight members across four generations identified a missense mutation in the histone deacetylase 4 (HDAC4) gene that segregated with illness. ESRRA and HDAC4 were determined to interact both in vitro in HeLa cells and in vivo in mouse cortex. Transcriptional analysis revealed that HDAC4 potently represses the expression of known ESRRA-induced target genes. Biochemical analysis of candidate mutations revealed that the identified ESRRA mutation decreased its transcriptional activity, while the HDAC4 mutation increased transcriptional repression of ESRRA. Our findings suggest that mutations that result in decreased ESRRA activity increase the risk of developing EDs.


Partial heat acclimation in cricketers using a 4-day high intensity cycling protocol.
Cricketers are often required to play in hot/humid environments with little time for heat adaptation. We examined the effect of a short 4-d hot/humid acclimation program on classical physiological indicators of heat acclimation. Male club cricketers were randomly assigned into heat acclimation (ACC, n = 6) or control (CON, n = 6) groups, and 30 min treadmill trials (10 km/h, approx. 30 ± 1.0°C, approx. 65 ± 6% RH) were conducted at baseline and postacclimation. The ACC group completed four high intensity (30-45 min) acclimation sessions on consecutive days at approx. 30°C and approx. 60% RH using a cycle ergometer. The CON group completed matched cycle training in moderate conditions (approx. 20°C, approx. 60% RH). Physiological measures during each treadmill trial included heart rate; core and skin temperatures; sweat Na+, K+ and Cl- electrolyte concentrations; and sweat rate. After the 4-d intervention, the ACC group had a moderate decrease of -11 (3 to -24 beats/min; mean and 90% CI) in the 30 min heart rate, and moderate to large reductions in electrolyte concentrations: Na+ -18% (-4 to -31%), K+ -15% (0 to -27%), Cl- -22% (-9 to -33%). Both ACC and CON groups had only trivial changes in core and skin temperatures and sweat rate. After the intervention, both groups perceived they were more comfortable exercising in the heat. The 4-d heat intervention had no detrimental effect on performance. Four 30-45 min high intensity cycle sessions in hot/humid conditions elicited partial heat acclimation. For full heat acclimation a more intensive and extensive (and modality-specific) acclimation intervention is needed for cricket players.


WE-G-217BCD-11: A New Phantom to Study Combined Effects of In-Plane (x,y) and Z Axis Resolution for 3-D Imaging.
The aim of this work is to develop phantoms that can be used to sample the radial and 3D properties of a CT image, including in-plane (x,y) and z-axis information. The Phantom is amenable to mathematical analysis of the x, y, and z axis resolution properties separately and combined. A periodic pattern of a pair of opposed (30°) angled ramps is configured to produce a waveform profile across the CT image. A perfect CT image (with no loss of resolution) of the test object would produce a consistent geometric pattern of the intersection of a line with the pair of angled ramps. However, due to the finite resolution (x, y and z), the CT waveform profile will not yield the perfect profile; rather it will be influenced by slice thickness, and in-plane resolution (PSF, MTF), as well as noise limitations, and other sources of non-uniformity such as beam hardening etc. Various characteristics of the waveform profile including, amplitude, frequency, and slope (rate of climb) of the peaks, can be studied using mathematical analysis such as the Fourier transform. It will be shown how these performance characteristics are encoded in the wave pattern. The waveform profiles are visually examined and mathematically analyzed, to demonstrate the effect of Slice Thickness (z axis) and changes of In-Plane (x,y) Resolution and non-uniformity across the image field; moreover, the harmonic analysis of the waveform is used to predict, either the in-plane resolution (MTF), or the z-axis MTF when one of the two is already known. The Wave pattern phantom offers a way to consider 3-D imaging characteristics of a CT scanner by scanning a single repetitive test object that encodes both in-plane resolution and z-axis resolution and also offers a way to study non-uniformity effects throughout the CT plane (volume). DJG is a consultant to The Phantom Laboratory and Image OWL, Salem, NY. Funding of other authors is supplied by Image OWL Salem, NY.


RK2, a glial-specific homeodomain protein required for embryonic nerve cord condensation and viability in Drosophila.
We report the identification of RK2, a glial-specific homeodomain protein. RK2 is localized to the nucleus of virtually all embryonic and imaginal glial cells, with the exception of midline glia. Embryos mutant for the gene encoding RK2 are embryonic lethal but normal for early gliogenesis (birth, initial divisions and migration of glia) and axonogenesis (neuronal pathfinding and fasciculation). However, later in development, there are significantly fewer longitudinal glia that are spatially disorganized; in addition, there is a slight disorganization of axon fascicles and a defective nerve cord condensation. This suggests that RK2 is not required for early glial determination, but rather for aspects of glial differentiation or function that are required for embryonic viability.


Impact of SARS on avian influenza preparedness in healthcare workers.
SARS was an unprecedented outbreak which brought about 1,755 infections and 302 deaths in Hong Kong. The similarity of SARS and avian influenza prompted us to examine the relationship between SARS experience and preparedness on a potential avian influenza outbreak. A self-administered questionnaire was delivered to nurses in Hong Kong to assess their attitude towards avian influenza; risk perception, and their relationships with previous level of exposure to SARS patients. Nine hundred and ninety-nine respondents were included in data analysis. About half of them perceived there would be an avian influenza outbreak in Hong Kong. The majority accepted a personal risk of infection in the course of their work (72.7%), and prepared to take care of patients infected with avian influenza (84.0%). Respondents were classified into two groups: high exposure (44.1%) and low exposure (55.9%) as defined by having worked in SARS ward or hospitals. High exposure nurses were less likely to avoid patients, less inclined to change their job if they were required to take care of infected patients, and had therefore a more positive attitude towards an impending avian influenza epidemic. About half of the nurses had frequent involuntary recalls of incidents relating to SARS, the frequency of which was positively correlated with knowing a person suffering from long-term complications of SARS. Healthcare workers who had been actively involved in SARS work were more "positive" in responding to the impending avian influenza epidemic. Whether the level of preparedness can be sustained would need to be further explored.


Treatment of Fordyce spots with CO2 laser.
Fordyce spots are heterotopic sebaceous glands that can be located at the lips' vermilion or the oral mucosa. Although this is considered a rather common disorder, a treatment for this condition that sometimes affects patients from only a cosmetic viewpoint has not yet been described. To evaluate CO2 superpulsed laser treatment in two subjects with Fordyce spots. Two patients with papules and yellowish plaques at the upper lip corresponding to Fordyce spots were treated with coherent Ambulase CO2 superpulsed laser (Coherent Medical, Palo Alto, CA); after informed consent was obtained, two to three passes were performed in one session using 2 and 4 W and a spot size of 2 mm. Complete re-epithelization was observed 2 weeks later with no residual Fordyce papules in the treated area and no side effects. Our findings suggest that CO2 superpulsed laser can be considered a safe and effective treatment for patients with Fordyce spots, offering excellent cosmetic results.


Prolonged Response to Anti-PD-1 Antibody Therapy in Chemotherapy-Refractory Cholangiocarcinoma With High Tumor Mutational Burden.
Management of advanced intrahepatic cholangiocarcinoma (iCCA) is challenging and overall survival is poor. Progress in the development of new therapeutic options for metastatic cholangiocarcinoma (CCA) has been slow; hence, to date, there are no approved second-line agents in this setting. Although the development of immune checkpoint inhibitors has significantly improved overall survival in a variety of malignancies, there has not been a clinically important impact in CCA. This report presents a 66-year-old patient with chemotherapy-refractory iCCA who experienced a prolonged response to immunotherapy. Tumor genome profiling revealed a high tumor mutation burden of 17 mutations per megabase in the absence of microsatellite instability. He was started on immunotherapy with nivolumab and has experienced an ongoing response for 16 months without clinical symptoms and only minimal radiologic disease.


Night calls: an Irish dimension.
Night calls made by 17 general practitioners in Cork in the Republic of Ireland in 1980 were analysed and compared with similar studies in the UK. There were 323 night calls: 11 per cent were classified as irresponsible, 59 per cent of which involved patients in social classes IV and V who make up 26.5 per cent of the population of Cork. Young children and the elderly had more calls than expected, and significant sex differences emerged in all age groups apart from the 45-65 age group. The incidence of night calls done comes towards the lower range of studies in the UK.


HIV, aging and continuity care: strengthening health systems to support services for noncommunicable diseases in low-income countries.
Although health systems in most low-income countries largely provide episodic care for acute symptomatic conditions, many HIV programs have developed effective, locally owned and contextually appropriate policies, systems and tools to support chronic care services for persons living with HIV (PLWH). The continuity of care provided by such programs may be especially critical for older PLWH, who are at risk for more rapid progression of disease and are more likely to have complications of HIV and its treatment than their younger counterparts. Older PLWH are also more likely to have other chronic noncommunicable diseases (NCDs), including hypertension, diabetes, cancers and chronic lung disease. As the number of older PLWH rises, enhanced chronic care systems will be required to optimize their health and wellbeing. These systems, lessons and resources can also be leveraged to support the burgeoning numbers of HIV-negative individuals with chronic NCD in need of ongoing care.


Abnormal auditory mismatch response in tinnitus sufferers with high-frequency hearing loss is associated with subjective distress level.
Tinnitus is an auditory sensation frequently following hearing loss. After cochlear injury, deafferented neurons become sensitive to neighbouring intact edge-frequencies, guiding an enhanced central representation of these frequencies. As psychoacoustical data 123 indicate enhanced frequency discrimination ability for edge-frequencies that may be related to a reorganization within the auditory cortex, the aim of the present study was twofold: 1) to search for abnormal auditory mismatch responses in tinnitus sufferers and 2) relate these to subjective indicators of tinnitus. Using EEG-mismatch negativity, we demonstrate abnormalities (N = 15) in tinnitus sufferers that are specific to frequencies located at the audiometrically normal lesion-edge as compared to normal hearing controls (N = 15). Groups also differed with respect to the cortical locations of mismatch responsiveness. Sources in the 90-135 ms latency window were generated in more anterior brain regions in the tinnitus group. Both measures of abnormality correlated with emotional-cognitive distress related to tinnitus (r approximately .76). While these two physiological variables were uncorrelated in the control group, they were correlated in the tinnitus group (r =.72). Concerning relationships with parameters of hearing loss (depth and slope), slope turned out to be an important variable. Generally, the steeper the hearing loss is the less distress related to tinnitus was reported. The associations between slope and the relevant neurophysiological variables are in agreement with this finding. The present study is the first to show near-to-complete separation of tinnitus sufferers from a normal hearing control group based on neurophysiological variables. The finding of lesion-edge specific effects and associations with slope of hearing loss corroborates the assumption that hearing loss is the basis for tinnitus development. It is likely that some central reorganization follow a damage to hearing receptors, even though the paradoxical results indicate that they most likely are somewhat different than originally assumed (see Background). One partial explanation might lie in the involvement of top-down (presumably frontal-lobe) controlled processes. A better comprehension of the exact mechanisms leading to the present results could have a broad impact on the understanding and perhaps treatment of tinnitus.


Maternal immune markers in serum during gestation and in breast milk and the risk of asthma-like symptoms at ages 6 and 12 months: a longitudinal study.
The role of breast milk on the risk of childhood asthma is in dispute. The aim of this prospective study is to determine the relationship of immune markers in maternal serum during gestation and breast milk to asthma-like symptoms (AS) in infancy. Pregnant women were recruited in Columbia and Charleston, South Carolina. Blood (median: three weeks before delivery) and breast milk (three weeks after delivery) samples were collected. Concentrations of interferon (IFN)-γ, IFN gamma-induced protein 10 (IP-10 or CXCL10), CCL11, interleukin (IL) 1β, IL-4, IL-5, IL-6, CXCL8, IL-10, IL-12(p70), IL-13, transforming growth factor (TGF)-β1, and immunoglobulin (Ig) A in both maternal serum and milk whey were determined via immunoassays. Asthma-like symptoms (AS) of the infant were ascertained at 6 and 12 months, respectively. Generalized estimating equations assessed relative risks (RRs) of immune markers for repeated measurements of AS, considering intra-individual correlations and adjusting for confounders. To provide comparable risk estimates, quartiles of the immune markers were used, except for IL-5 in whey and IgA in serum, which were dichotomized. Of 178 women, 161 provided blood and 115 breast milk samples. IL-12(p70), IL-4, IL-10, IL-1β, and CCL11 in serum and in whey were not further considered for the statistical analyses since the proportion of non-detectable values was high. Most immune markers in serum and milk whey were moderately or highly correlated; however, IgA was negatively correlated. Infants in the highest quartile of IL-13 in both serum and whey were at a higher risk of AS (RR = 3.02 and 4.18; respectively) compared to infants in the first quartile. High levels of IL-5 in serum and whey was also identified as a risk. In addition, increased secretory IgA and TGF-β1 in breast milk reduced the risks of AS. Maternal serum and whey levels of IL-5 and IL-13 are risk markers for AS; whey IgA and TGF-β1 seem to be protective. Only focusing on breast milk portend that milk cytokines IL-5 and IL-13 have adverse effects. However, similar immune exposures during late gestation and via milk suggest that both may enhance AS among infants.


Genome mapping with anchored clones: theoretical aspects.
As part of our effort to construct a physical map of the genome of Arabidopsis thaliana we have made a mathematical analysis of our experimental approach of anchoring yeast artificial chromosome clones with genetically mapped RFLPs and RAPDs. The details of this analysis are presented and their implications for mapping the Arabidopsis genome are discussed.


In vitro and in vivo experimental models for drug screening and development for Chagas disease.
Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.


Role of membrane lipids in the mechanism of bacterial species selective toxicity by two alpha/beta-antimicrobial peptides.
We have previously shown that two synthetic antimicrobial peptides with alternating alpha- and beta-amino acid residues, designated simply as alpha/beta-peptide I and alpha/beta-peptide II, had toxicity toward bacteria and affected the morphology of bacterial membranes in a manner that correlated with their effects on liposomes with lipid composition similar to those of the bacteria. In the present study we account for the weak effects of alpha/beta-peptide I on liposomes or bacteria whose membranes are enriched in phosphatidylethanolamine (PE) and why such membranes are particularly susceptible to damage by alpha/beta-peptide II. The alpha/beta-peptide II has marked effects on unilamellar vesicles enriched in PE causing vesicle aggregation and loss of their internal aqueous contents. The molecular basis of these effects is the ability of alpha/beta-peptide II to induce phase segregation of anionic and zwitterionic lipids as shown by fluorescence and differential scanning calorimetry. This phase separation could result in the formation of defects through which polar materials could pass across the membrane as well as form a PE-rich membrane domain that would not be a stable bilayer. alpha/beta-Peptide II is more effective in this regard because, unlike alpha/beta-peptide I, it has a string of two or three adjacent cationic residues that can interact with anionic lipids. Although alpha/beta-peptide I can destroy membrane barriers by converting lamellar to non-lamellar structures, it does so only weakly with unilamellar vesicles or with bacteria because it is not as efficient in the aggregation of these membranes leading to the bilayer-bilayer contacts required for this phase conversion. This study provides further understanding of why alpha/beta-peptide II is more toxic to micro-organisms with a high PE content in their membrane as well as for the lack of toxicity of alpha/beta-peptide I with these cells, emphasizing the potential importance of the lipid composition of the cell surface in determining selective toxicity of anti-microbial agents.


Interventions for treating headlice.
Infection with head lice is a widespread condition in developed and developing countries. Infection occurs most commonly in children, but also affects adults. If left untreated the condition can become intensely irritating and skin infections may occur if the bites are scratched. The aim of this review was to assess the effects of interventions for head lice. Trials register of The Cochrane Infectious Diseases Group; Medline; Embase; Science Citation Index; Biosis and Toxline; reference lists of relevant articles; pharmaceutical companies producing pediculicides (published and unpublished trials); UK and US Regulatory Authorities. Randomised trials (published and unpublished) or trials using alternate allocation were sought which compared pediculicides with the same and different formulations of other pediculicides, and pediculicides with physical methods. Of the 71 identified studies, only four met the inclusion criteria. Two reviewers independently assessed trial quality. One reviewer extracted the data. We found no evidence that any one pediculicide has greater effect than another. The two studies comparing malathion and permethrin with their respective vehicles showed a higher cure rate for the active ingredient than the vehicle. Another study comparing synergised pyrethrins with permethrin showed their effects to be equivalent. A comparative trial of malathion lotion vs combing, showed combing to be ineffective for the cureative treatment of head lice infection. Adverse effects were reported in a number of trials and were all minor, although reporting quality varied between trials. Permethrin, synergised pyrethrin and malathion were effective in the treatment of head lice. However, the emergence of drug resistance since these trials were conducted means there is no direct contemporary evidence of the comparative effectiveness of these products. The 'best' choice will now depend on local resistance patterns. Physical treatment methods(BugBusting) were shown ot be ineffective to treat head lice. No evidence exists regarding other chemical control methods such as the use of herbal treatments, when used in the curative treatment of head lice. Future trials should take into account the methodological recommendations that arise from this review.


Four super water-stable lanthanide-organic frameworks with active uncoordinated carboxylic and pyridyl groups for selective luminescence sensing of Fe(3.).
Four super water-stable isostructural lanthanide-organic frameworks, [Ln(HL)(H2O)2]n·2H2O (1-Ln) (Ln = Eu, Tb, Nd and Sm), have been successfully synthesized under hydrothermal conditions with 3,5-di(2,4-dicarboxylphenyl)pyridine (H4L) and Ln(NO3)3·6H2O. 1-Ln shows a novel 2D layered structure with uncoordinated carboxylic and pyridyl groups pointing to the interior of interlayer channels. The luminescent properties of 1-Eu in the solid state and one steeped in aqueous solution have been measured, which show excellent luminescence and good luminescent stability in water. Next 1-Eu was chosen as a probe for sensing different metal ions. Consequently, 1-Eu exhibits a highly selective response to Fe(3+) through the luminescence quenching effect in aqueous solutions. The probable mechanisms of the quenching effects have been investigated in detail.


A decline in prosocial language helps explain public disapproval of the US Congress.
Talking about helping others makes a person seem warm and leads to social approval. This work examines the real world consequences of this basic, social-cognitive phenomenon by examining whether record-low levels of public approval of the US Congress may, in part, be a product of declining use of prosocial language during Congressional debates. A text analysis of all 124 million words spoken in the House of Representatives between 1996 and 2014 found that declining levels of prosocial language strongly predicted public disapproval of Congress 6 mo later. Warm, prosocial language still predicted public approval when removing the effects of societal and global factors (e.g., the September 11 attacks) and Congressional efficacy (e.g., passing bills), suggesting that prosocial language has an independent, direct effect on social approval.


Toxoplasma gondii infection and risk of Parkinson and Alzheimer diseases: A systematic review and meta-analysis on observational studies.
In the past decade, Toxoplasma gondii infection has been recognized as a potential risk for many psychiatric and neurological disorders. We performed this systematic review and meta-analysis to evaluate the association between Toxoplasma infection and Parkinson's and Alzheimer's diseases. PubMed, Web of science, Scopus and Embase databases were searched up to September 30, 2018 for studies that reported risk of Parkinson's and Alzheimer's diseases associated with Toxoplasma infection. We used a random effects meta-analysis model to generate the pooled odds ratio (OR) with 95% confidence intervals (CIs). Eleven studies, including seven studies for Parkinson's disease (428 patients and 540 controls) and four studies for Alzheimer's disease (301 patients and 313 controls), were included in the meta-analysis. We found that there was no statistically significant association between Toxoplasma infection, as determined by IgG serology, IgM serology, and PCR with increased risk of Parkinson's disease (OR, 1.14; 95% CI, 0.78-1.68), (OR, 1.61; 95% CI, 0.33-7.76) and (OR, 1.87; 95% CI, 0.43-8.05), respectively. The OR for association of Toxoplasma infection, based on IgG serology with Alzheimer's patients, compared to control group, was (OR, 1.38; 95% CI, 0.99-1.92), demonstrating a marginally significant association between Toxoplasma infection and Alzheimer's disease. Our findings do not support a general hypotheses regarding an associative relationship between Toxoplasma infection and Parkinson's disease, but do support a marginally significant association between Toxoplasma infection and Alzheimer's disease; this association should be investigated further through longitudinal and experimental studies.


A model for the clustered distribution of SNPs in the human genome.
Motivated by a non-random but clustered distribution of SNPs, we introduce a phenomenological model to account for the clustering properties of SNPs in the human genome. The phenomenological model is based on a preferential mutation to the closer proximity of existing SNPs. With the Hapmap SNP data, we empirically demonstrate that the preferential model is better for illustrating the clustered distribution of SNPs than the random model. Moreover, the model is applicable not only to autosomes but also to the X chromosome, although the X chromosome has different characteristics from autosomes. The analysis of the estimated parameters in the model can explain the pronounced population structure and the low genetic diversity of the X chromosome. In addition, correlation between the parameters reveals the population-wise difference of the mutation probability. These results support the mutational non-independence hypothesis against random mutation.


A multiple-SNP approach for genome-wide association study of milk production traits in Chinese Holstein cattle.
The multiple-SNP analysis has been studied by many researchers, in which the effects of multiple SNPs are simultaneously estimated and tested in a multiple linear regression. The multiple-SNP association analysis usually has higher power and lower false-positive rate for detecting causative SNP(s) than single marker analysis (SMA). Several methods have been proposed to simultaneously estimate and test multiple SNP effects. In this research, a fast method called MEML (Mixed model based Expectation-Maximization Lasso algorithm) was developed for simultaneously estimate of multiple SNP effects. An improved Lasso prior was assigned to SNP effects which were estimated by searching the maximum joint posterior mode. The residual polygenic effect was included in the model to absorb many tiny SNP effects, which is treated as missing data in our EM algorithm. A series of simulation experiments were conducted to validate the proposed method, and the results showed that compared with SMMA, the new method can dramatically decrease the false-positive rate. The new method was also applied to the 50k SNP-panel dataset for genome-wide association study of milk production traits in Chinese Holstein cattle. Totally, 39 significant SNPs and their nearby 25 genes were found. The number of significant SNPs is remarkably fewer than that by SMMA which found 105 significant SNPs. Among 39 significant SNPs, 8 were also found by SMMA and several well-known QTLs or genes were confirmed again; furthermore, we also got some positional candidate gene with potential function of effecting milk production traits. These novel findings in our research should be valuable for further investigation.


An ion for an iron: streptococcal metal homeostasis under oxidative stress.
The ability of opportunistic pathogens such as Group A Streptococcus (GAS) to transition between mucosal colonisation and invasive disease requires complex systems for adapting to markedly different host environments. The battle to acquire essential trace metals such as manganese and iron from the host is central to pathogenesis. Using a molecular genetic approach, Turner et al. [Biochem. J. (2019) 476, 595-611] show that it is not just individual metal concentrations that are important, but the ratio of iron to manganese within cells. Increasing this ratio by knocking out pmtA, encoding the Fe(II) exporter PmtA, or by disrupting mtsA, encoding an MtsABC Mn(II)-import system component, led to reductions in superoxide dismutase (SodA) activity and increased sensitivity to oxidative stress. The authors show that SodA is at least 4-fold more active with Mn bound than with Fe and speculate that high intracellular Fe:Mn ratios reduce superoxide dismutase activity through the mismetalation of SodA. Challenging wild-type GAS with 1 mM H2O2 led to a decrease in Fe:Mn ratio and a 3-fold increase in SodA activity, indicating that modulation of the balance between intracellular Fe and Mn may play an important role in adaptation to oxidative stress. This work unravels some of the key mechanisms for maintaining appropriate Mn and Fe concentrations within bacterial cells and underscores the need for future studies that take an holistic view to metal ion homeostasis in bacteria. Strategies aimed at interfering with the balance of intracellular metal ions represent a promising approach for the control of invasive microbial infections.


Preferences for very low and very high voice pitch in humans.
Manipulations of voice pitch have been shown to alter attractiveness ratings, but whether preferences extend to very low or very high voice pitch is unknown. Here, we manipulated voice pitch in averaged men's and women's voices by 2 Hz intervals to create a range of male and female voices speaking monopthong vowel sounds and spanning a range of frequencies from normal to very low and very high pitch. With these voices, we used the method of constant stimuli to measure preferences for voice. Nineteen university students (ages: 20-25) participated in three experiments. On average, men preferred high-pitched women's voices to low-pitched women's voices across all frequencies tested. On average, women preferred men's voices lowered in pitch, but did not prefer very low men's voices. The results of this study may reflect selection pressures for men's and women's voices, and shed light on a perceptual link between voice pitch and vocal attractiveness.


Effects of wind energy development on nesting ecology of greater prairie-chickens in fragmented grasslands.
Wind energy is targeted to meet 20% of U.S. energy needs by 2030, but new sites for development of renewable energy may overlap with important habitats of declining populations of grassland birds. Greater Prairie-Chickens (Tympanuchus cupido) are an obligate grassland bird species predicted to respond negatively to energy development. We used a modified before-after control-impact design to test for impacts of a wind energy development on the reproductive ecology of prairie-chickens in a 5-year study. We located 59 and 185 nests before and after development, respectively, of a 201 MW wind energy facility in Greater Prairie-Chicken nesting habitat and assessed nest site selection and nest survival relative to proximity to wind energy infrastructure and habitat conditions. Proximity to turbines did not negatively affect nest site selection (β = 0.03, 95% CI = -1.2-1.3) or nest survival (β = -0.3, 95% CI = -0.6-0.1). Instead, nest site selection and survival were strongly related to vegetative cover and other local conditions determined by management for cattle production. Integration of our project results with previous reports of behavioral avoidance of oil and gas facilities by other species of prairie grouse suggests new avenues for research to mitigate impacts of energy development.


Polymorphic expression of UDP-glucuronosyltransferase UGTlA gene in human colorectal cancer.
Polymorphism of genes encoding drug-metabolizing enzymes is known to play an important role in increased susceptibility of colorectal cancer. UGT1A gene locus has been suggested to define tissue-specific glucuronidation activity. Reduced capacity of glucuronidation is correlated with the development of colorectal cancer. Therefore, we sought to explore polymorphism of UGTlA gene in human colorectal cancer. Cancerous and healthy tissues were obtained from selectedpatients. Blood samples were collected and UGTlA mRNA transcriptions were analyzed. Genomic DNA was prepared and UGTlA8 exon-1 sequences were amplified, visualized and purified. The extracted DNA was subcloned and sequenced. Two-tailed Fisher's exact test, Odds ratios (ORs), confidence interval (CIs) and Logistics Regression Analysis were used for statistical analysis. UGTlA mRNA expression was reduced in cancerous tissues compared with healthy tissues from the same patient . The UGTlA mRNA expression of healthy tissue in study patients was lower than control . The mRNA expression of cancerous tissue was down-regulated in UGTlAl, 1A3, 1A4, lA6, 1A9 and up-regulated in UGTlA8 and UGTlAl0 UGT1A5 and UGT1A7 were not expressed in colonic tissue of either group. The allele frequency of WT UGTlA8*1 was higher (p = 0.000), frequency of UGTlA8*3 was lowered in control group (p = 0.000). The expression of homozygous UGTlA8*1 was higher in control group (p = 0.000). Higher frequency of both heterozygous UGTlA8*1/*3 and UGTlA8*2/*3 were found in study group (p = 0.000; p = 0.000). The occurrence of colorectal cancer was mainly related to the presence of polymorphic UGTlA8*3 alleles (p = 0.000). Regulation of human UGT1A genes is tissue-specific. Individual variation in polymorphic expressions of UGTlA gene locus was noted in all types of colonic tissue tested, whereas hepatic tissue expression was uniform. The high incidence of UGTlA8 polymorphism exists in colorectal cancer patients. UGTlA8*1 allele is a protective factor and UGTlA8*3 allele is a risk factor.


The Biomechanical Consequences of Trapeziectomy and Partial Trapezoidectomy in the Treatment of Thumb Carpometacarpal and Scaphotrapeziotrapezoid Arthritis.
To determine, using a biomechanical cadaveric model, whether, in the treatment of thumb carpometacarpal and scaphotrapeziotrapezoid arthritis, partial trapezoid resection following trapeziectomy causes carpal, specifically lunocapitate and scapholunate, instability. Eight fresh-frozen mid-forearm cadaver specimens with type I lunates and devoid of basilar thumb arthritis were used in the study. Specimens were mounted onto a wrist simulator applying cyclical wrist flexion/extension and radial/ulnar deviation motions. Carpal kinematics, specifically lunocapitate and scapholunate joint relationships, were measured at 4 different conditions: (1) a native intact state, (2) after trapeziectomy, (3) after 2-mm partial trapezoid resection, and (4) after 4-mm partial trapezoid resection. During both flexion/extension and radial/ulnar deviation of the wrist, the lunocapitate and scapholunate joint relationship did not show any notable change following any of trapeziectomy, 2-mm, or 4-mm trapezoid resection compared with the intact state. Changes to the lunocapitate and scapholunate angles were clinically insignificant-a maximum of 6° and 4° change, respectively. This biomechanical cadaveric study shows that performing a trapeziectomy followed by up to 4 mm of proximal trapezoid resection has a negligible effect upon carpal, specifically lunocapitate and scapholunate, stability. Further research is needed to elucidate the long-term clinical consequences of limited trapezoid resection in vivo. There may be no clinically relevant effects of resection of up to 4 mm of trapezoid in the surgical management of combined basilar thumb and scaphotrapeziotrapezoid arthritis.


A Prebiotic Formula Improves the Gastrointestinal Bacterial Flora in Toddlers.
We aimed to investigate the effect of enriched 3-prebiotic formula (including inulin, fructooligosaccharides, and galactooligosaccharides) on toddler gut health by measuring fecal microbiota. Our results revealed that the consumption of 3-prebiotic formula three times per day giving total intake of 1.8 g prebiotic ingredients significantly showed the increased number of probiotic Bifidobacterium spp. colonies and the reduced populations of both C. perfringens and total anaerobic bacteria on the fecal bacterial flora in toddlers at 18~36 months. In addition, total organic acids in the fecal samples significantly increased which improves the utilization of bifidus under acidic conditions after consumption of the 3-prebiotic formula. Therefore, using the formula enriched with prebiotic may maintain gut health in toddlers.


The anonymity paradox in patient engagement: reputation, risk and web-based public feedback.
The UK National Health Service (NHS) has long espoused patient and public engagement. Recent years have seen increasing use of internet-based methods of collecting feedback about patient experience and public and staff views about NHS services and priorities. Often hailed as a means of facilitating participative democratic patient engagement, these processes raise a number of complex issues. A key aspect of it is the opportunity for comment to be made anonymously. Our research reveals an anonymity paradox whereby patients clearly demonstrate a perception that anonymity is a prerequisite for effective use of these feedback processes, whereas professionals demonstrate a perception that patient anonymity is a barrier to effective use. The risks of anonymity are constructed very differently by patients and professionals. Patient concerns around anonymity were not motivated by a general concern about a loss of privacy, but more that a positive identification might compromise future care. For professionals, concerns were voiced more around risks of reputational damage for specific practitioners or practices (in that anyone could say anything) and also that this anonymous feedback was available publicly and that it might go against the medical opinion of the professional. These concerns pointed to important differences in perceptions of patient and professional vulnerability. In the qualitative analysis that follows the key finding was that while anonymity makes service users feel less vulnerable, it can have the opposite effect on managers and clinical staff. This raises important implications for the use and utility of internet-based methods of collecting patient feedback.


Effect of food insecurity on chronic kidney disease in lower-income Americans.
The relation of food insecurity (inability to acquire nutritionally adequate and safe foods) and chronic kidney disease (CKD) is unknown. We examined whether food insecurity is associated with prevalent CKD among lower-income individuals in both the general US adult population and an urban population. We conducted cross-sectional analyses of lower-income participants of the National Health and Nutrition Examination Survey (NHANES) 2003-2008 (n = 9,126) and the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study (n = 1,239). Food insecurity was defined based on questionnaires and CKD was defined by reduced estimated glomerular filtration rate or albuminuria; adjustment was performed with multivariable logistic regression. In NHANES, the age-adjusted prevalence of CKD was 20.3, 17.6, and 15.7% for the high, marginal, and no food insecurity groups, respectively. Analyses adjusting for sociodemographics and smoking status revealed high food insecurity to be associated with greater odds of CKD only among participants with either diabetes (OR = 1.67, 95% CI: 1.14-2.45 comparing high to no food insecurity groups) or hypertension (OR = 1.37, 95% CI: 1.03-1.82). In HANDLS, the age-adjusted CKD prevalence was 5.9 and 4.6% for those with and without food insecurity, respectively (p = 0.33). Food insecurity was associated with a trend towards greater odds of CKD (OR = 1.46, 95% CI: 0.98-2.18) with no evidence of effect modification across diabetes, hypertension, or obesity subgroups. Food insecurity may contribute to disparities in kidney disease, especially among persons with diabetes or hypertension, and is worthy of further study.


Palliative care for non-cancer conditions in primary care: a time trend analysis in the UK (2009-2014).
While guidelines recommend palliative care in non-cancer conditions, this has not been widely implemented. We examined whether the recording of a palliative care approach and the numbers of hospital deaths for deceased patients with heart failure, dementia, chronic obstructive pulmonary disease (COPD) and cancer have changed since the UK End-of-Life Care Strategy was introduced. We conducted sequential cross-sectional studies of decedents within the UK's Clinical Practice Research Datalink and Hospital Episode Statistics. All adults with a primary care record of COPD (n=5426), dementia (n=7339), heart failure (n=6409) or cancer (n=18 668) who died during three 1 year periods (April 2009 to March 2014) were included. Evidence of a palliative care approach was identified from primary care records, and death in hospital from secondary care data. From 2009 to 2014, proportions with a primary care record of palliative care increased for COPD from 13.6% to 21.2%; dementia from 20.9% to 40.7%; and heart failure from 12.6% to 21.2%; but remained substantially lower than for cancer (57.6% to 61.9%). Median days before death of recording improved for COPD (145 to 224) and dementia (44 to 209); but not for heart failure (168.5 to 153) and cancer (123 to 114). Trends in hospital deaths were not consistently downward, although the proportions of patients dying in hospital were lower in the last period compared with the first. Recording of a palliative care approach for non-cancer conditions has increased since the introduction of the UK End-of-Life Care Strategy, but remains inadequate.


Motor Vehicle Crash Risk Among Adolescents and Young Adults With Attention-Deficit/Hyperactivity Disorder.
Attention-deficit/hyperactivity disorder (ADHD) often persists into adolescence, when motor vehicle crash risk peaks. We know little about when adolescents with ADHD get licensed and, once they do, the extent to which they have increased crash risk compared with adolescents without ADHD. To examine the association between ADHD and both driver licensing and crash involvement and whether it varies by sex, licensing age, and/or being prescribed ADHD medication at licensure. This retrospective cohort study was conducted at 6 primary care practices of the Children's Hospital of Philadelphia, a large pediatric health care network in southeastern Pennsylvania and southern New Jersey. Using electronic health records, we defined a cohort of 2479 adolescents and young adults with ADHD and 15 865 without ADHD who were (1) born from 1987 to 1997; (2) residents of New Jersey and patients at 1 of 6 New Jersey primary care practices at age 12 years or older; and (3) age-eligible to obtain a driver's license from 2004 through 2014. Electronic health records data were then linked with New Jersey's statewide driver licensing and crash databases for 2004 through 2014. Acquisition of a driver's license and first involvement as a driver in a police-reported crash. Survival analysis was used to estimate adjusted hazard ratios for licensing and crash outcomes through age 25 years. The median age of individuals at the end of the study was 22.2 years (interquartile range, 19.7-24.8). Compared with individuals without ADHD, the licensing probability of individuals with ADHD 6 months after eligibility was 35% lower (for males: adjusted hazard ratio, 0.65; 95% CI, 0.61-0.70; females: adjusted hazard ratio, 0.64; 95% CI, 0.58-0.70). Among individuals with a driver's license, 764 of 1785 with ADHD (42.8%) and 4715 of 13 221 without ADHD (35.7%) crashed during the study period. The adjusted risk for first crash among licensed drivers with ADHD was 1.36 times higher than for those without ADHD (95% CI, 1.25-1.48) and did not vary by sex, licensing age, or over time. Only 129 individuals with ADHD (12.1%) were prescribed medication in the 30 days before licensure. Adolescents with ADHD get licensed less often and at an older age. Once licensed, this cohort has a greater risk of crashing. Additional research is needed to understand the specific mechanisms by which ADHD influences crash risk.


Circulating catecholamines in exercise and hyperventilation induced asthma.
Plasma noradrenaline, adrenaline, and cyclic 3'5' AMP (cAMP) were measured in seven asthmatic patients with known exercise-induced bronchospasm and six matched non-atopic control subjects during a standard treadmill exercise test and then during matched isocapnic hyperventilation. Normal subjects showed a 5.5 fold rise in noradrenaline and a 3.2 fold rise in adrenaline during exercise compared with a 2.1 fold rise in noradrenaline and no significant rise in adrenaline in asthmatics who all developed bronchoconstriction after exercise (mean fall in peak flow rate 28.4 +/- 5.8%). Plasma cAMP rose 1.4 fold in controls but showed no significant rise in asthmatics. This reduced sympatho-adrenal response to exercise in asthmatics is difficult to explain. The failure of circulating catecholamines to rise and stimulate beta adrenoceptors on the mast cell may facilitate the release of bronchoconstrictor mediators. Matched hyperventilation produced bronchospasm in asthmatics (mean fall in peak flow rate 29.0 +/- 4.4%) but no change in catecholamines in either group suggesting that circulating catecholamines have no direct role in exercise-induced bronchospasm but may play a permissive role via the mast cell.


Reversing the pipeline? Implementing public health evidence-based guidance in english local government.
In the UK, responsibility for many public health functions was transferred in 2013 from the National Health Service (NHS) to local government; a very different political context and one without the NHS history of policy and practice being informed by evidence-based guidelines. A problem this move presented was whether evidence-based guidelines would be seen as relevant, useful and implementable within local government. This study investigates three aspects of implementing national evidence-based recommendations for public health within a local government context: influences on implementation, how useful guidelines are perceived to be and whether the linear evidence-guidelines-practice model is considered relevant. Thirty-one councillors, public health directors and deputy directors and officers and other local government employees were interviewed about their experiences implementing evidence-based guidelines. Interviews were informed and analysed using a theoretical model of behaviour (COM-B; Capability, Opportunity, Motivation-Behaviour). Contextual issues such as budget, capacity and political influence were important influences on implementation. Guidelines were perceived to be of limited use, with concerns expressed about recommendations being presented in the abstract, lacking specificity and not addressing the complexity of situations or local variations. Local evidence was seen as the best starting point, rather than evidence-based guidance produced by the traditional linear 'evidence-guidelines-practice' model. Local evidence was used to not only provide context for recommendations but also replace recommendations when they conflicted with local evidence. Local government users do not necessarily consider national guidelines to be fit for purpose at local level, with the consequence that local evidence tends to trump evidence-based guidelines. There is thus a tension between the traditional model of guideline development and the needs of public health decision-makers and practitioners working in local government. This tension needs to be addressed to facilitate implementation. One way this might be achieved, and participants supported this approach, would be to reverse or re-engineer the traditional pipeline of guideline development by starting with local need and examples of effective local practice rather than starting with evidence of effectiveness synthesised from the international scientific literature. Alternatively, and perhaps in addition, training about the relevance of research evidence should become a routine for local government staff and councillors.


Clinical characteristics and long-term visual outcome of severe phenotypes of chronic central serous chorioretinopathy.
The aim of this study was to investigate disease onset and disease progression in patients with severe chronic central serous chorioretinopathy (cCSC). The medical records of 143 cCSC patients (199 eyes) were reviewed. All cases had visual complaints for >6 months and showed signs of a severe disease phenotype on optical coherence tomography (OCT) and fluorescein angiography (FA). Clinical presentation at onset was evaluated, together with disease progression on multimodal imaging and final treatment outcome. Twenty-eight cases (14%) had a documented history of an acute episode of CSC, whereas 145 cases (73%) showed pre-existing features of chronicity already at first presentation. The first clinical presentation could not be evaluated in 13% of cases. Best-corrected visual acuity (BCVA) was 70 ± 18 Early Treatment of Diabetic Retinopathy Study (ETDRS) letters at onset and 70 ± 22 ETDRS letters at final visit (p = 0.770). Among all studied cases, 173 eyes (87%) were treated, which resulted in complete resolution of subretinal fluid (SRF) in 76% of eyes at final visit. In eyes with fluorescein angiographic follow-up, the area of diffuse atrophic retinal pigment epithelium (RPE) abnormalities (diffuse atrophic RPE alterations [DARA]) had increased significantly in 43 eyes (68%) at final visit. CSC encompasses a clinical spectrum that includes a range of severe phenotypes, in which retinal abnormalities tend to be progressive. Nevertheless, the long-term visual acuity may remain fairly stable with treatment. Few patients with severe chronic CSC have a history of acute CSC, which could indicate that there may be pathogenetic differences between these 2 CSC variants.


Differential effects of dihydropyridine calcium channel blockers in kainic acid-induced experimental seizures in rats.
The anticonvulsant effects of the dihydropyridine calcium channel blockers nifedipine, nicardipine and nimodipine were studied on experimental seizures induced by intra-hippocampal injection of kainic acid (KA) in chloralhydrate anesthetized Wistar rats. The rats were anesthetized and placed in a stereotaxic apparatus. After midline incision four screw electrodes were placed over the left and right frontal and parietal cortex and KA was injected into left dorsal hippocampus via 5-microliter Hamilton microsyringe. The changes in electroencephalograph (EEG) activity and EEG power spectra were recorded in basal conditions and 5, 10, 15, 30 and 60 min following KA injection. KA-induced excitatory changes in the surface EEG activity were associated with the marked increase in EEG power spectra in the frequency range from 14.5-22 Hz. Pretreatment with nifedipine, nicardipine and nimodipine revealed that they exerted certain differences in their anticonvulsant properties. Nimodipine significantly delayed the onset of seizures and prevented the KA-induced changes in EEG and in EEG power spectra in all recorded channels and in a dose dependent manner. Nifedipine exerted significant anticonvulsant effect only in channel four, while nicardipine was ineffective. Our results suggest that anticonvulsant action of some dihydropyridine calcium channel blockers, especially nimodipine may be in part independent of its antagonism on L-type voltage-gated calcium (Ca(2+)) channels.


Encoding of motion targets by waves in turtle visual cortex.
Visual stimuli evoke wave activity in the visual cortex of freshwater turtles. Earlier work from our laboratory showed that information about the positions of stationary visual stimuli is encoded in the spatiotemporal dynamics of the waves and that the waves can be decoded using Bayesian detection theory. This paper extends these results in three ways. First, it shows that flashes of light separated in space and time and stimuli moving with three speeds can be discriminated statistically using the waves generated in a large-scale model of the cortex. Second, it compares the coding capabilities of spike rate and spike time codes. Spike rate codes were obtained by low-pass filtering the activities of individual neurons in the model with filters of different band widths. For the moving targets used in the study, detectability using spike rate codes is immune to the choice of a specific bandwidth, indicating that a coarse filter is able to adequately discriminate targets. Spike timing codes are binary sequences indicating the precise timing of spike activity of individual neurons across the cortex. Spike time codes generally perform better than do spike rate codes. Third, the encoding process is examined in terms of the underlying cellular mechanisms that result in the initiation, propagation and cessation of the wave. The period of peak detectability corresponds to the period in which waves are propagating across the cortex.


Integrative medicine: a tale of two clinics.
Integrative medicine (blending the best of complementary and alternative medicine (CAM) with conventional medicine) is becoming increasingly popular. The objectives of this paper are to compare and contrast the development of two teams that set out to establish integrative medical clinics, highlighting key issues found to be common to both settings, and to identify factors that appear to be necessary for integration to occur. At St Michael's Hospital (an inner-city teaching hospital in Toronto, Canada), a total of 42 interviews were conducted between February 2004 and August 2006 wi18 key participants (4 administrators, 2 chiropractors, 2 physiotherapists and 10 family physicians). At the CARE (Complementary and Alternative Research and Education) Program at Stollery Children's Hospital, Edmonton, Canada, 44 interviews were conducted with 24 people on four occasions: June 2004, March 2005, November 2006, and June 2007. Basic content analysis was used to identify the key themes from the transcribed interviews. Despite the contextual differences between the two programs, a striking number of similar themes emerged from the data. The five most important shared themes were: 1) the necessity of "champions" and institutional facilitators to conceive of, advocate for, and bring the programs to fruition; 2) the credibility of these champions and facilitators (and the credibility of the program being established) was key to the acceptance and growth of the program in each setting; 3) the ability to find the "right" practitioners and staff to establish the integrative team was crucial to each program's ultimate success; 4) the importance of trust (both the trustworthiness of the developing program as well as the trust that developed between the practitioners in the integrative team); and 5) the challenge of finding physical space to house the programs. The programs were ultimately successful because of the credibility of the champions, institutional facilitators and the staff members. Selection of excellent clinicians who were able to work well as a team facilitated the establishment of trust both within the team itself as well as between the team and the host institution.


Klotho, a gene related to a syndrome resembling human premature aging, functions in a negative regulatory circuit of vitamin D endocrine system.
The klotho gene encodes a novel type I membrane protein of beta-glycosidase family and is expressed principally in distal tubule cells of the kidney and choroid plexus in the brain. These mutants displayed abnormal calcium and phosphorus homeostasis together with increased serum 1,25-(OH)2D. In kl-/- mice at the age of 3 wk, elevated levels of serum calcium (10.9 +/- 0.31 mg/dl vs. 10.0 +/- 0.048 mg/dl in wild-type mice), phosphorus (14.7 +/- 1.1 mg/dl vs. 9.7 +/- 1.5 mg/dl in wild type) and most notably, 1,25-(OH)2D (403 +/- 99.7 mg/dl vs. 88.0 +/- 34.0 mg/dl in wild type) were observed. Reduction of serum 1,25-(OH)2D concentrations by dietary restriction resulted in alleviation of most of the phenotypes, suggesting that they are downstream events resulting from elevated 1,25-(OH)2D. We searched for the signals that lead to up-regulation of vitamin D activating enzymes. We examined the response of 1alpha-hydroxylase gene expression to calcium regulating hormones, such as PTH, calcitonin, and 1,25-(OH)2D3. These pathways were intact in klotho null mutant mice, suggesting the existence of alternate regulatory circuits. We also found that the administration of 1,25-(OH)2D3 induced the expression of klotho in the kidney. These observations suggest that klotho may participate in a negative regulatory circuit of the vitamin D endocrine system, through the regulation of 1alpha-hydroxylase gene expression.


Spinal cord trauma and the molecular point of no return.
A mechanical trauma to the spinal cord can be followed by the development of irreversible and progressive neurodegeneration, as opposed to a temporary or partially reversible neurological damage. An increasing body of experimental and clinical evidence from humans and animal models indicates that spinal cord injury may set in motion the development of disabling and at times fatal neuromuscular disorders, whose occurrence is not normally associated with any major environmental event. This outcome appears to be dependent on the co-occurrence of a particular form of mechanical stress and of a genetically-determined vulnerability. This increased vulnerability to spinal cord injury may depend on a change of the nature and of the timing of activation of a number of neuroprotective and neurodestructive molecular signals in the injured cord. Among the main determinants, we could mention an altered homeostasis of lipids and neurofilaments, an earlier inflammatory response and the failure of the damaged tissue to rein in oxidative damage and apoptotic cell death. These changes could force injured tissue beyond a point of no return and precipitate an irreversible neurodegenerative process. A better knowledge of the molecular signals activated in a state of increased vulnerability to trauma can inform future treatment strategies and the prediction of the neurological outcome after spinal cord injury.


Serum CA19.9 levels are commonly elevated in primary ovarian mucinous tumours but cannot be used to predict the histological subtype.
CA19.9 is a monosialoganglioside secreted by mucinous tumours of the gastrointestinal tract, including the pancreas and biliary tree. Limited studies have shown that this tumour marker may also be elevated in primary ovarian mucinous neoplasms, but no study has assessed whether serum CA19.9 levels can be used to predict if a primary ovarian mucinous tumour is benign, borderline or malignant. The aim of this study was to correlate the serum CA19.9 level with the histological features in a large series of primary ovarian mucinous neoplasms. 144 cases of primary ovarian mucinous neoplasm (79 benign, 45 borderline and 20 malignant) were identified in which a preoperative serum CA19.9 level had been performed. The association between the serum levels and the histological subtype and a variety of other parameters was investigated. In a subset of cases, immunohistochemical staining for CA19.9 was performed on tumour blocks. Serum CA19.9 levels were elevated in 27%, 38% and 40% of mucinous cystadenomas, borderline mucinous tumours and mucinous carcinomas, respectively. Markedly elevated levels of serum CA19.9 were observed in each group, with the highest serum CA19.9 measurements being noted in borderline mucinous tumours. There was no relationship between the serum CA19.9 level and whether the tumours were benign, borderline or malignant (Kruskal-Wallis test p value=0.32). A weak but statistically significant correlation was found between tumour maximum dimension and CA19.9 level (Spearman's rank correlation coefficient=0.17, p=0.04). In those cases in which CA19.9 immunohistochemistry was performed, all tumours showed positive staining for CA19.9, with 60% of these cases being associated with an elevated serum CA19.9 level. Preoperative CA19.9 levels cannot be used to predict whether a suspected ovarian mucinous tumour is benign, borderline or malignant. Markedly elevated serum levels (>1000 U/ml) may be found in benign mucinous neoplasms as well as in borderline and malignant tumours.


A Drosophila model of multiple endocrine neoplasia type 2.
Dominant mutations in the Ret receptor tyrosine kinase lead to the familial cancer syndrome multiple endocrine neoplasia type 2 (MEN2). Mammalian tissue culture studies suggest that RetMEN2 mutations significantly alter Ret-signaling properties, but the precise mechanisms by which RetMEN2 promotes tumorigenesis remain poorly understood. To determine the signal transduction pathways required for RetMEN2 activity, we analyzed analogous mutations in the Drosophila Ret ortholog dRet. Overexpressed dRetMEN2 isoforms targeted to the developing retina led to aberrant cell proliferation, inappropriate cell fate specification, and excessive Ras pathway activation. Genetic analysis indicated that dRetMEN2 acts through the Ras-ERK, Src, and Jun kinase pathways. A genetic screen for mutations that dominantly suppress or enhance dRetMEN2 phenotypes identified new genes that are required for the phenotypic outcomes of dRetMEN2 activity. Finally, we identified human orthologs for many of these genes and examined their status in human tumors. Two of these loci showed loss of heterozygosity (LOH) within both sporadic and MEN2-associated pheochromocytomas, suggesting that they may contribute to Ret-dependent oncogenesis.


Periorbital lymphatic malformation: clinical course and management in 42 patients.
Lymphatic malformation in the orbital cavity and surrounding region often causes disfigurement and visual problems. To better clarify the evolution and treatment of this condition, the authors studied a retrospective cohort of 42 consecutive patients seen between 1971 and 2003 and analyzed anatomic features, complications, and management. The ratio of female to male patients was 1:1. Most periorbital lymphatic malformations were noted at birth (59 percent), presenting as either unilateral swelling (60 percent) or a periorbital mass (24 percent). Sixty-two percent of lesions were on the left side. The ipsilateral cheek, temple, and forehead also were involved in 57 percent of patients. Twenty-two percent of lesions were intraconal, 30 percent were extraconal, and 48 percent were in both spaces. Forty-five percent of children had an associated cerebral developmental venous anomaly. Periorbital lymphatic malformation caused major morbidity; 52 percent of patients had intralesional bleeding and 26 percent of patients had a history of infection. Other common complications included intermittent swelling (76 percent), blepharoptosis (52 percent), proptosis (45 percent), pain (21 percent), amblyopia (33 percent), chemosis (19 percent), astigmatism (17 percent), and strabismus (7 percent). Ultimately, 40 percent of children had diminished vision and 7 percent became blind in the affected eye. Management of periorbital lymphatic malformation involved an interdisciplinary team that included an interventional radiologist, a craniofacial surgeon, and an ophthalmologist. The two therapeutic strategies were sclerotherapy (40 percent) and resection (57 percent); most patients required several interventions. A coronal approach was used for subtotal excision of fronto-temporal-orbital lymphatic malformation in 13 patients, whereas a tarsal incision was used for lesions isolated to the eyelid (n = 14). Ocular proptosis was temporarily managed by tarsorrhaphy (n = 9), but expansion of the bony orbit was needed to correct persistent proptosis (n = 8). Orbital exenteration was necessary in two patients.