The bactericidal effect of human secreted group IID phospholipase A2 results from both hydrolytic and non-hydrolytic activities.
The Human Secreted Group IID Phospholipase A(2) (hsPLA2GIID) may be involved in the human acute immune response. Here we have demonstrated that the hsPLA2GIID presents bactericidal and Ca(2+)-independent liposome membrane-damaging activities and we have compared these effects with the catalytic activity of active-site mutants of the protein. All mutants showed reduced hydrolytic activity against DOPC:DOPG liposome membranes, however bactericidal effects against Escherichia coli and Micrococcus luteus were less affected, with the D49K mutant retaining 30% killing of the Gram-negative bacteria at a concentration of 10μg/mL despite the absence of catalytic activity. The H48Q mutant maintained Ca(2+)-independent membrane-damaging activity whereas the G30S and D49K mutants were approximately 50% of the wild-type protein, demonstrating that phospholipid bilayer permeabilization by the hsPLA2GIID is independent of catalytic activity. We suggest that this Ca(2+)-independent damaging activity may play a role in the bactericidal function of the protein.


The effect of rotator cuff tears on reaction forces at the glenohumeral joint.
The rotator cuff muscles maintain glenohumeral stability by compressing the humeral head into the glenoid. Disruption of the rotator cuff compromises concavity compression and can directly affect the loads on the glenohumeral joint. The purpose of this study was to quantify the effect of rotator cuff tears on the magnitude and direction of glenohumeral joint reaction forces during active shoulder abduction in the scapular plane using nine cadaveric upper extremities. Motion of the full upper extremity was simulated using a dynamic shoulder testing apparatus. Glenohumeral joint reaction forces were measured by a universal force-moment sensor. Five conditions of rotator cuff tears were tested: Intact, Incomplete Supraspinatus Tear, Complete Supraspinatus Tear, Supraspinatus/Infraspinatus Tear, and Global Tear. Reaction forces at the glenohumeral joint were found to steadily increase throughout abduction and peaked at maximum abduction for all conditions tested. There were no significant differences in reaction force magnitude for the intact condition (337 +/- 88 N) or those involving an isolated incomplete tear (296 +/- 83 N) or complete tear (300 +/- 85 N) of the supraspinatus tendon. Extension of tears beyond the supraspinatus tendon into the anterior and posterior aspect of the rotator cuff led to a significant decrease in the magnitude of joint reaction force (126 +/- 31 N). Similarly, such tears resulted in a significant change in the direction of the reaction force at the glenohumeral joint. These results suggest that joint reaction forces are significantly affected by the integrity of the rotator cuff, specifically, by the transverse force couple formed by the anterior and posterior aspects of the cuff. The quantitative data obtained in this study on the effect of rotator cuff tears on magnitude and direction of the reaction force at the glenohumeral joint helps clarify the relationship between joint motion, joint compression and stability.


Semantic memory impairment for biological and man-made objects in individuals with amnestic mild cognitive impairment or late-life depression.
Amnestic mild cognitive impairment (aMCI) and late-life depression (LLD) both increase the risk of developing Alzheimer disease (AD). Very little is known about the similarities and differences between these syndromes. The present study addresses this issue by examining the nature of semantic memory impairment (more precisely, object-based knowledge) in patients at risk of developing AD. Participants were 17 elderly patients with aMCI, 18 patients with aMCI plus depressive symptoms (aMCI/D+), 15 patients with LLD, and 29 healthy controls. All participants were aged 55 years or older and were administered a semantic battery designed to assess semantic knowledge for 16 biological and 16 man-made items. Overall performance of aMCI/D+ participants was significantly worse than the 3 other groups, and performance for questions assessing knowledge for biological items was poorer than for questions relating to man-made items. This study is the first to show that aMCI/D+ is associated with object-based semantic memory impairment. These results support the view that semantic deficits in aMCI are associated with concomitant depressive symptoms. However, depressive symptoms alone do not account exclusively for semantic impairment, since patients with LLD showed no semantic memory deficit.


Effects of sunlight on bacteriophage viability and structure.
Current estimates of viral abundance in natural waters rely on direct counts of virus-like particles (VLPs), using either transmission or epifluorescence microscopy. Direct counts of VLPs, while useful in studies of viral ecology, do not indicate whether the observed VLPs are capable of infection and/or replication. Rapid decay in bacteriophage viability under environmental conditions has been observed. However, it has not been firmly established whether there is a corresponding degradation of the virus particles. To address this question, viable and direct counts were carried out employing two Chesapeake Bay bacteriophages in experimental microcosms incubated for 56 h at two depths in the York River estuary. Viruses incubated in situ in microcosms at the surface yielded decay rates in full sunlight of 0.11 and 0.06 h-1 for CB 38 phi and CB 7 phi, respectively. The number of infective particles in microcosms in the dark and at a depth of 1 m was not significantly different from laboratory controls, with decay rates averaging 0.052 h-1 for CB 38 phi and 0.037 h-1 for CB 7 phi. Direct counts of bacteriophages decreased in teh estuarine microcosms, albeit only at a rate of 0.028 h-1, and were independent of treatment. Destruction of virus particles is concluded to be a process separate from loss of infectivity. It is also concluded that strong sunlight affects the viability of bacteriophages in surface waters, with the result that direct counts of VLPs overestimate the number of bacteriophage capable of both infection and replication. However, in deeper waters, where solar radiation is not a significant factor, direct counts should more accurately estimate numbers of viable bacteriophage.


Impact of prophylactic beta-blocker therapy to prevent stroke after noncardiac surgery.
beta Blockers are widely used to improve the postoperative cardiac outcome in patients with coronary artery disease scheduled for noncardiac surgery. However, recently serious concerns regarding the safety of perioperative beta blockers have emerged. To assess the incidence, risk factors, and beta-blocker use associated with postoperative stroke in the Dutch Echocardiographic Cardiac Risk Evaluation Applying Stress Echocardiography (DECREASE) trials, we evaluated all 3,884 patients of the DECREASE trials for postoperative stroke. All cardiac risk factors and medication use were assessed. The incidence of stroke within 30 days after surgery was recorded. The incidence of postoperative stroke in the DECREASE trials was 0.46% (18 of 3,884). For the beta-blocker users, the incidence was 0.5%. All the strokes had an ischemic origin. A history of stroke was associated with a greater incidence of postoperative stroke (odds ratio [OR] 3.79, 95% confidence interval [CI] 1.2 to 11.6). Statins and anticoagulants were not associated with postoperative stroke (OR 0.85, 95% CI 0.3 to 2.4; and OR 1.27, 95% CI 0.4 to 4.6, respectively). No association with bisoprolol therapy was found (OR 1.16, 95% CI 0.4 to 3.4). In conclusion, with a low-dose bisoprolol regimen started > or =30 days before surgery, no association was observed between beta-blocker use and postoperative stroke.


Automated method for determining the flow of surface functionalized nanoparticles through a hydraulically fractured mineral formation using plasmonic silver nanoparticles.
Quantifying nanoparticle (NP) transport within porous geological media is imperative in the design of tracers and sensors to monitor the environmental impact of hydraulic fracturing that has seen increasing concern over recent years, in particular the potential pollution and contamination of aquifers. The surface chemistry of a NP defining many of its solubility and transport properties means that there is a wide range of functionality that it is desirable to screen for optimum transport. Most prior transport methods are limited in determining if significant adsorption occurs of a NP over a limited column distance, however, translating this to effects over large distances is difficult. Herein we report an automated method that allows for the simulation of adsorption effects of a dilute nanoparticle solution over large distances under a range of solution parameters. Using plasmonic silver NPs and UV-visible spectroscopic detection allows for low concentrations to be used while offering greater consistency in peak absorbance leading to a higher degree of data reliability and statistics. As an example, breakthrough curves were determined for mercaptosuccinic acid (MSA) and cysteamine (CYS) functionalized Ag NPs passing through Ottawa sand (typical proppant material) immobile phase (C) or bypassing the immobile phase (C0). Automation allows for multiple sequences such that the absorption plateau after each breakthrough and the rate of breakthrough can be compared for multiple runs to provide statistical analysis. The mobility of the NPs as a function of pH is readily determined. The stickiness (α) of the NP to the immobile phase calculated from the C/C0 ratio shows that MSA-Ag NPs show good mobility, with a slight decrease around neutral pH, while CYS-Ag NPs shows an almost sinusoidal variation. The automated process described herein allows for rapid screening of NP functionality, as a function of immobile phase (proppant versus reservoir material), hydraulic fracturing fluid additives (guar, surfactant) and conditions (pH, temperature).


Urinary sodium excretion has positive correlation with activation of urinary renin angiotensin system and reactive oxygen species in hypertensive chronic kidney disease.
It is not well described the pathophysiology of renal injuries caused by a high salt intake in humans. The authors analyzed the relationship between the 24-hr urine sodium-to-creatinine ratio (24HUna/cr) and renal injury parameters such as urine angiotensinogen (uAGT/cr), monocyte chemoattractant peptide-1 (uMCP1/cr), and malondialdehyde-to-creatinine ratio (uMDA/cr) by using the data derived from 226 hypertensive chronic kidney disease patients. At baseline, the 24HUna/cr group or levels had a positive correlation with uAGT/cr and uMDA/cr adjusted for related factors (P<0.001 for each analysis). When we estimated uAGT/cr in the 24HUna/cr groups by ANCOVA, the uAGT/cr in patients with ≥200 mEq/g cr was higher than in patients with <100 mEq/g cr (708 [95% CI, 448-967] vs. 334 [95% CI, 184-483] pg/mg cr, P=0.014). Similarly, uMDA/cr was estimated as 0.17 (95% CI, 0.14-0.21) pM/mg cr in patients with <100 mEq/g cr and 0.27 (95% CI, 0.20-0.33) pM/mg cr in patients with ≥200 mEq/g cr (P=0.016). During the 16-week follow-up period, an increase in urinary sodium excretion predicted an increase in urinary angiotensinogen excretion. In conclusion, high salt intake increases renal renin-angiotensin-system (RAS) activation, primarily, and directly or indirectly affects the production of reactive oxygen species through renal RAS activation.


Candidate causal regulatory effects by integration of expression QTLs with complex trait genetic associations.
The recent success of genome-wide association studies (GWAS) is now followed by the challenge to determine how the reported susceptibility variants mediate complex traits and diseases. Expression quantitative trait loci (eQTLs) have been implicated in disease associations through overlaps between eQTLs and GWAS signals. However, the abundance of eQTLs and the strong correlation structure (LD) in the genome make it likely that some of these overlaps are coincidental and not driven by the same functional variants. In the present study, we propose an empirical methodology, which we call Regulatory Trait Concordance (RTC) that accounts for local LD structure and integrates eQTLs and GWAS results in order to reveal the subset of association signals that are due to cis eQTLs. We simulate genomic regions of various LD patterns with both a single or two causal variants and show that our score outperforms SNP correlation metrics, be they statistical (r(2)) or historical (D'). Following the observation of a significant abundance of regulatory signals among currently published GWAS loci, we apply our method with the goal to prioritize relevant genes for each of the respective complex traits. We detect several potential disease-causing regulatory effects, with a strong enrichment for immunity-related conditions, consistent with the nature of the cell line tested (LCLs). Furthermore, we present an extension of the method in trans, where interrogating the whole genome for downstream effects of the disease variant can be informative regarding its unknown primary biological effect. We conclude that integrating cellular phenotype associations with organismal complex traits will facilitate the biological interpretation of the genetic effects on these traits.


Impact of transgenic oilseed rape expressing oryzacystatin-1 (OC-1) and of insecticidal proteins on longevity and digestive enzymes of the solitary bee Osmia bicornis.
The risk that insect-resistant transgenic plants may pose for solitary bees was assessed by determining longevity of adult Osmia bicornis (O. rufa) chronically exposed to transgenic oilseed rape expressing oryzacystatin-1 (OC-1) or to the purified insecticidal proteins recombinant rOC-1, Kunitz soybean trypsin inhibitor (SBTI), Galanthus nivalis agglutinin (GNA), or Bacillus thuringiensis toxin Cry1Ab dissolved in sugar solution (at 0.01 and 0.1%, w:v, Cry1Ab only at 0.01%). Compared to control bees, longevity was significantly reduced by SBTI and GNA at both concentrations and by rOC-1 at 0.1%, but not by Cry1Ab or rOC-1 at 0.01%. Longevity on the OC-1 oilseed rape was not significantly different from the control plants. The effects of SBTI and rOC-1 on longevity were investigated through characterization of the digestive proteinases of O. bicornis and analysis of the response in proteinase profiles to ingestion of these proteinase inhibitors. A relatively complex profile of at least four types of soluble proteolytic enzymes was identified. Serine proteinases were found to be predominant, with metallo and especially cysteine proteinases making a smaller albeit significant contribution. The compensatory response to in vivo enzyme inhibition was similar for SBTI and rOC-1 although less pronounced for rOC-1. It consisted of a non-specific overproduction of native proteinases, both sensitive and insensitive, and the induction of a novel aspartic proteinase.


Image plane detector for Fabry-Perot interferometers: physical model and improvement with anticoincidence detection.
Modern Fabry-Perot interferometers use image plane array detection to obtain the multiplex advantage. The imaging quality of these image plane detectors (IPD's) limits their ultimate resolution and usefulness. The influence of pulse spreading on photon-counting IPD's is investigated, first theoretically and then experimentally on real devices. The model developed in our study is in good agreement with laboratory measurements and suggests a more reliable technique to measure the blurring of the Fabry-Perot fringe caused by the IPD. Based on the model, an anticoincidence detection circuit is designed; the circuit is simple, greatly reduces the blurring, and increases the resolution of the IPD.


Occupational noise exposure and hearing loss: A study of knowledge, attitude and practice among Tanzanian iron and steel workers.
We assessed Knowledge Attitude and Practice (KAP) regarding occupational noise exposure, Noise-induced hearing loss, audiometry and use of hearing protection devices among iron and steel factory workers exposed to high noise level. A modified, validated, structured questionnaire was used to collect information from 253 male workers randomly selected from the four factories. The sum scores for each domain of KAP were computed. Scores above 75% were defined as good knowledge and positive attitude. For practice, scores of >50% were defined as good. Independent samples t-test and Chi-squared test were used to analyze association between KAP and continuous/categorical variables respectively. Majority of workers displayed poor knowledge and poor practice (94%), but 76% displayed a positive attitude. Most of the workers (86%) had never been provided with hearing protection devices. The mean scores for attitude and practice differed significantly between the four factories (one-way ANOVA, p < 0.001). Implementation of hearing conservation program with provision of hearing protection devices are suggested.


Thoracic aortic aneurysm repair with endovascular stent-grafts.
The purpose of the study was to describe the clinical experience is using endoluminal stent-grafts for the treatment of thoracic aortic aneurysms in high-risk patients. Patients with aneurysms of the descending thoracic aorta who were considered high surgical risks underwent evaluation for endoluminal repair. The prosthesis was constructed from Z stents covered with polyester fabric using dimensions based upon preprocedural computed tomography scans and angiography. Through a femoral arteriotomy or left retroperitoneal flank incision, a 22-24 Fr delivery catheter was inserted and advanced through the aorta to the target site under fluoroscopic guidance in the operating suite. The stent-graft prosthesis was deployed at the site of the aneurysm. 44 patients (36 male, 8 female; mean age 36 years) underwent stent-graft repair for thoracic aneurysms (mean diameter 6.3 cm). The deployment was technically successful in all cases, with complete aneurysm thrombosis in 88%. The 30-day perioperative mortality rate was 6.8% and 35-month actuarial survival was 82%. There were no cases of stent migration, surgical conversion or intraprocedural death. Paraplegia occurred in two patients who underwent simultaneous surgical infrarenal aortic aneurysm repair immediately followed by stent-graft placement for a coexisting thoracic aneurysm. The conclusion was that placement of endoluminal stent-grafts for repair of thoracic aortic aneurysms is technically feasible in high-risk patients in whom conventional surgery is contraindicated. Long-term studies are needed to determine protection against aneurysm rupture and patient survival.


Methylphenidate and atomoxetine prescribing trends in children in the Western Cape Province of South Africa, 2005-2013.
There is paucity of data on methylphenidate and atomoxetine prescribing patterns in South African children. To describe the prescribing trends of these agents in children residing in the Western Cape Province. South African private health sector. Longitudinal drug utilisation study on medicine claims data from 2005-2013, focussing on the number of patients and prescriptions per patient. The total number of patients increased by 29.5% from 2005 to 2013. The majority were boys (male:female ratio, 3.5:1), and between the ages of > 6 and ≤12 years in 2005 and >12 and ≤18 years in 2013. More than 75% of patients received methylphenidate or atomoxetine in the City of Cape Town Metropolitan municipality. Prescriptions for methylphenidate and atomoxetine increased by 45.5% overall from 2005 to 2013 (p < 0.001), with that for methylphenidate and atomoxetine increasing by 36.0% and 314.5%, respectively. The average number of annual methylphenidate prescriptions per patient increased from 3.96 ± 2.92 (95% CI, 3.69-4.23) in 2005 to 4.38 ± 2.85 (95% CI, 4.14-4.61) in 2013 (Cohen's d = 0.14) and for atomoxetine from 2.58 ± 1.86 (95% CI, 1.80-3.37) in 2005 to 4.85 ± 3.66 (95% CI, 3.84-5.86) in 2013 (Cohen's d = 0.62). Although the total number of patients and prescribing of methylphenidate and atomoxetine increased significantly from 2005 to 2013, a slight downward trend was observed in the mean number of prescriptions per patient per year from 2008 onwards. These prescribing patterns warrant further research.


Methods for estimating human endogenous retrovirus activities from EST databases.
Human endogenous retroviruses (HERVs) are surviving traces of ancient retrovirus infections and now reside within the human DNA. Recently HERV expression has been detected in both normal tissues and diseased patients. However, the activities (expression levels) of individual HERV sequences are mostly unknown. We introduce a generative mixture model, based on Hidden Markov Models, for estimating the activities of the individual HERV sequences from EST (expressed sequence tag) databases. We use the model to estimate the relative activities of 181 HERVs. We also empirically justify a faster heuristic method for HERV activity estimation and use it to estimate the activities of 2450 HERVs. The majority of the HERV activities were previously unknown. (i) Our methods estimate activity accurately based on experiments on simulated data. (ii) Our estimate on real data shows that 7% of the HERVs are active. The active ones are spread unevenly into HERV groups and relatively uniformly in terms of estimated age. HERVs with the retroviral env gene are more often active than HERVs without env. Few of the active HERVs have open reading frames for retroviral proteins.


Assessment of cancer risks due to environmental exposure to asbestos.
In a rural area widespread pollution of friable and non-friable waste products was present, used to harden dirt tracks, yards, and driveways during 1935-1974. Exposure to environmental asbestos was assessed by a site approach, based on number of polluted sites within postal code areas, and by a household approach, based on number of households in the close vicinity to polluted sites within postal code areas. Based on asbestos soil investigations, 293 sites were identified with asbestos waste material at the surface, of which 77% contained crocidolite fibres as well as chrysotile fibres. The 293 sites-at-risk varied from 5 m(2) to 2722 m(2) and were surrounded by 347 households within 100 m of these sites. Distance to the plant was associated with the number of sites (r=0.36), and with the number of households (r=0.52). However, categorization of postal code areas into low, intermediate or high likelihood of exposure to asbestos showed a modest agreement between the site and household approach. In the site approach a total of 2.3 million person-years at risk were estimated with an average exposure of 1674 fibres/m(3) and an expected 1.8 cases of malignant mesothelioma each year. The household approach resulted in estimates of 1.2 million person-years at risk, and 0.9 cases of malignant mesothelioma per year, respectively. This study illustrates that asbestos waste on the surface of roads and yards in an area with over 130,000 inhabitants may result in long-term exposure to asbestos that will cause several cases of malignant mesothelioma each year. Although distance to plant, number of polluted sites and number of exposed household were associated, the modest agreement among these measures of exposure indicate that the exposure assessment strategy chosen in a particular study may result in considerable misclassification. Without detailed information on individual behaviour within the polluted area, it is difficult to show that a more individually oriented approach will perform better than an ecological approach.


Marchiafava-Bignami disease: literature review and case report.
We postulated that disruption of callosal pathways as occurs in Marchiafava-Bignami disease (MBD) is associated with marked impairment in brain functioning as measured by cognitive assessment and neuroimaging. MBD is considered to be a rare and severe complication of chronic alcoholism. It is characterized by necrosis and subsequent atrophy of the corpus callosum, which is the major brain structure connecting corresponding areas of both hemispheres. We review the existing literature on MBD with respect to conceptualization, theories of pathogenesis, forms of the disease, and neuroimaging and neuropsychological findings. We then present the case of a middle-aged man with MBD who underwent extensive clinical, neuropsychological, and neuroimaging studies. Neuropsychological evaluation revealed a pattern of severe global dementia. Magnetic resonance imaging showed moderate atrophy of anterior callosal regions and severe atrophy of posterior callosal regions in the setting of cortical and subcortical atrophy. Resting metabolism positron emission tomography revealed decreased glucose metabolism most pronounced in subcortical and mesial frontal regions. The differential diagnosis, function of the corpus callosum, and potential limitations of our case study are discussed. On account of the history, clinical presentation, and results of magnetic resonance imaging of the brain, we diagnosed our patient with chronic MBD.


Decline of forest interior conditions in the conterminous United States.
Forest fragmentation threatens the sustainability of forest interior environments, thereby endangering subordinate ecological attributes and functions. We analyzed the spatial patterns of forest loss and gain for the conterminous United States from 2001 to 2006 to determine whether forest interior environments were maintained at five spatial scales. A 1.1% net loss of total forest area translated to net losses of 3.2% to 10.5% of forest interior area over spatial scales of 4.41 ha to 5,310 ha. At the 65.6-ha scale, the reduction of forest interior area was 50,000 km²--almost double the net loss of total forest area. The pervasive discrepancy between total forest loss and forest interior loss indicates a widespread shift of the extant forest to more fragmented conditions, even in regions exhibiting small net changes in extant forest area. In the conterminous United States, trends in total forest area underestimate threats to forest from forest fragmentation.


Breed-specific ancestry studies and genome-wide association analysis highlight an association between the MYH9 gene and heat tolerance in Alaskan sprint racing sled dogs.
Alaskan sled dogs are a genetically distinct population shaped by generations of selective interbreeding with purebred dogs to create a group of high-performance athletes. As a result of selective breeding strategies, sled dogs present a unique opportunity to employ admixture-mapping techniques to investigate how breed composition and trait selection impact genomic structure. We used admixture mapping to investigate genetic ancestry across the genomes of two classes of sled dogs, sprint and long-distance racers, and combined that with genome-wide association studies (GWAS) to identify regions that correlate with performance-enhancing traits. The sled dog genome is enhanced by differential contributions from four non-admixed breeds (Alaskan Malamute, Siberian Husky, German Shorthaired Pointer, and Borzoi). A principal components analysis (PCA) of 115,000 genome-wide SNPs clearly resolved the sprint and distance populations as distinct genetic groups, with longer blocks of linkage disequilibrium (LD) observed in the distance versus sprint dogs (7.5-10 and 2.5-3.75 kb, respectively). Furthermore, we identified eight regions with the genomic signal from either a selective sweep or an association analysis, corroborated by an excess of ancestry when comparing sprint and distance dogs. A comparison of elite and poor-performing sled dogs identified a single region significantly associated with heat tolerance. Within the region we identified seven SNPs within the myosin heavy chain 9 gene (MYH9) that were significantly associated with heat tolerance in sprint dogs, two of which correspond to conserved promoter and enhancer regions in the human ortholog.


An IAP retrotransposon in the mouse ADAMTS13 gene creates ADAMTS13 variant proteins that are less effective in cleaving von Willebrand factor multimers.
Severe deficiency of ADAMTS13, a von Willebrand factor (VWF)-cleaving metalloprotease, causes thrombotic thrombocytopenic purpura. When analyzed with VWF multimers, but not with an abbreviated VWF peptide (VWF73) as the substrate, the plasma ADAMTS13 activity levels of mouse strains segregated into a high and a low group that differed by approximately 10 fold. Low ADAMTS13 activity was detected in mice containing 2 alleles of intracisternal A-type particle (IAP) retrotransposon sequence in the ADAMTS13 gene. Molecular cloning of mouse ADAMTS13 identified 2 truncated variants (IAP-a and IAP-b) in the low-activity mice. Both of the IAP variants lacked the 2 carboxyl terminus thrombospondin type 1 repeat (TSR) and CUB domains of full-length ADAMTS13. The IAP-b variant also had splicing abnormalities affecting the spacer domain sequence and had miniscule enzymatic activity. Compared with full-length ADAMTS13, the IAP-a variant was approximately one ninth as active in cleaving VWF multimers but was only slightly less active in cleaving VWF73 peptide. Recombinant human ADAMTS13 was also less effective in cleaving VWF multimers than VWF73 when the C-terminal TSR sequence was deleted. In summary, the carboxyl terminus TSR sequence is important for cleaving VWF multimers. Assay results should be interpreted with caution when peptide substrates are used for analysis of variant ADAMTS13 proteins.


Bringing dicynodonts back to life: paleobiology and anatomy of a new emydopoid genus from the Upper Permian of Mozambique.
Dicynodontia represent the most diverse tetrapod group during the Late Permian. They survived the Permo-Triassic extinction and are central to understanding Permo-Triassic terrestrial ecosystems. Although extensively studied, several aspects of dicynodont paleobiology such as, neuroanatomy, inner ear morphology and internal cranial anatomy remain obscure. Here we describe a new dicynodont (Therapsida, Anomodontia) from northern Mozambique: Niassodon mfumukasi gen. et sp. nov. The holotype ML1620 was collected from the Late Permian K5 formation, Metangula Graben, Niassa Province northern Mozambique, an almost completely unexplored basin and country for vertebrate paleontology. Synchrotron radiation based micro-computed tomography (SRµCT), combined with a phylogenetic analysis, demonstrates a set of characters shared with Emydopoidea. All individual bones were digitally segmented allowing a 3D visualization of each element. In addition, we reconstructed the osseous labyrinth, endocast, cranial nerves and vasculature. The brain is narrow and the cerebellum is broader than the forebrain, resembling the conservative, "reptilian-grade" morphology of other non-mammalian therapsids, but the enlarged paraflocculi occupy the same relative volume as in birds. The orientation of the horizontal semicircular canals indicates a slightly more dorsally tilted head posture than previously assumed in other dicynodonts. In addition, synchrotron data shows a secondary center of ossification in the femur. Thus ML1620 represents, to our knowledge, the oldest fossil evidence of a secondary center of ossification, pushing back the evolutionary origins of this feature. The fact that the specimen represents a new species indicates that the Late Permian tetrapod fauna of east Africa is still incompletely known.


Dietary intakes consistent with the DASH dietary pattern reduce blood pressure increase with age and risk for stroke in a Chinese population.
The Dietary Approaches to Stop Hypertension (DASH) diet has been proven to effectively lower blood pressure(BP), and associate with a lower cardiovascular disease and stroke risk in mainly non-Asians. Further, it is unclear if adhering to the DASH target nutrients has similar BP impact as adhering to the recommended DASH food groups. Associations between adherence to DASH foods or nutrients and BP or stroke risk were assessed in 1420 and 2061 Taiwanese adults from 1989 to 2002, respectively. The DASH food score (p=0.053), dairy(p=0.030) and calcium (p=0.020) intake were significantly and inversely associated with follow up systolic BP change in univariate analyses. Both dairy (p=0.020) and calcium (p=0.001) also showed a consistent inverse association with systolic BP change in multivariate analysis. None of the factors examined was associated with diastolic BP change. Both DASH nutrient score and magnesium intakes were significantly associated with the hazard ratio (HR) for total stroke in an inverse relationship. The HR of total stroke comparing the highest to the lowest tertile was 0.63 (95% CI: 0.41-0.98, p=0.037) for the DASH nutrient score, and 0.62 (95% CI: 0.40-0.97,p=0.030) for magnesium intake. Similar findings were observed for DASH nutrient score (p=0.011) and magnesium intake (p=0.043) with the HR for ischemic stroke. The HR for total and ischemic stroke for calcium intake also showed a borderline trend (p=0.071 and 0.051, respectively). In conclusion, adhering to the DASH diet is beneficial for long term BP control and reduction of stroke risk in this Chinese population.


Infrared Nanospectroscopy Reveals DNA Structural Modifications upon Immobilization onto Clay Nanotubes.
The growing demand for innovative means in biomedical, therapeutic and diagnostic sciences has led to the development of nanomedicine. In this context, naturally occurring tubular nanostructures composed of rolled sheets of alumino-silicates, known as halloysite nanotubes, have found wide application. Halloysite nanotubes indeed have surface properties that favor the selective loading of biomolecules. Here, we present the first, to our knowledge, structural study of DNA-decorated halloysite nanotubes, carried out with nanometric spatially-resolved infrared spectroscopy. Single nanotube absorption measurements indicate a partial covering of halloysite by DNA molecules, which show significant structural modifications taking place upon loading. The present study highlights the constraints for the use of nanostructured clays as DNA carriers and demonstrates the power of super-resolved infrared spectroscopy as an effective and versatile tool for the evaluation of immobilization processes in the context of drug delivery and gene transfer.


Prevalence of Batrachochytrium dendrobatidis infection is extremely low in direct-developing Australian microhylids.
The emerging infectious disease chytridiomycosis has been implicated in declines and disappearances of amphibian populations around the world. However, susceptibility to infection and the extent of pathological effects of infection vary among hosts, and species with life histories that include parental care of direct-developing terrestrial eggs may tend to be less susceptible. We examined samples from a total of 595 individuals of 9 species of direct-developing Australian frogs in the family Microhylidae for the presence of infection by Batrachochytrium dendrobatidis (Bd). Between 1995 and 2004, 336 samples were collected; 102 of these were analysed histologically and 234 were tissues stored in alcohol, which were examined using diagnostic quantitative PCR (qPCR). Swab samples were collected from 259 frogs from 2005 to 2008 and were examined using qPCR. None of the 595 samples showed evidence of infection by Bd. If these data are regarded as a single sample representative of Australian microhylids, the upper 95% binomial confidence limit for the prevalence of infection in frogs of this family is 0.0062 (<1%). Even if only the data from the more powerful diagnostic qPCR tests are used, the upper 95% confidence limit for prevalence is 0.0075 (<1%). Our data suggest that Australian microhylids have a very low prevalence of infection by Bd in nature, and thus are either not susceptible, or are only slightly susceptible, to chytridiomycosis. This could be due solely to, or in combination with, low rates of transmission and to factors that promote resistance to infection, including ecological or behavioural characteristics, innate immune functions such as antimicrobial skin peptides, or antimicrobial symbionts in skin flora.


Insulin-responsive tissues contain the core complex protein SNAP-25 (synaptosomal-associated protein 25) A and B isoforms in addition to syntaxin 4 and synaptobrevins 1 and 2.
SNAP-25 (synaptosomal-associated protein 25), syntaxin and synaptobrevin are the three SNARE [soluble NSF attachment protein receptor (where NSF = N-ethylmaleimide-sensitive fusion protein)] proteins that form the core complex involved in synaptic vesicle docking and subsequent fusion with the target membrane. The present study is aimed at understanding the mechanisms of fusion of vesicles carrying glucose transporter proteins with the plasma membrane in human insulin-responsive tissues. It describes the isolation and characterization of cDNA molecules encoding SNAP-25 A and B isoforms, syntaxin 4 and synaptobrevins (also known as vehicle-associated membrane proteins) from two major human insulin-responsive tissues, skeletal muscle and fat. The DNA and deduced amino acid sequences of SNAP-25 revealed perfect identity with the previously reported human neural SNAP-25 A and B isoforms. Our results indicate the presence of both isoforms both in insulin-responsive tissues and in in vitro cultured 3T3-L1 cells, but suggest a differential pattern of gene expression: isoform A is the major species in adipose tissue, and isoform B is the major species in skeletal muscle. The presence of SNAP-25 protein in 3T3-L1 cells was demonstrated by immunofluorescence microscopy using an anti-SNAP-25 monoclonal antibody. Immunoprecipitation experiments using the same monoclonal antibody also revealed the presence of SNAP-25 protein in plasma membrane fractions from rat epididymal fat pads. The syntaxin 4-encoding region from skeletal muscle contains five nucleotide differences from the previously reported placental cDNA sequence, two of which result in amino acid changes: Asp-174 to Glu and Val-269 to Ala. The synaptobrevin 1 cDNA from skeletal muscle contains two nucleotide differences when compared with the corresponding clone from neural tissues, one of which is silent and the other resulting in the amino acid change Thr-102 to Ala. The cDNA sequence of the protein from fat is identical with that of human synaptobrevin 1 from neural tissues. Furthermore, we have confirmed the presence of syntaxin 4 in fat and of synaptobrevin 2 in skeletal muscle by PCR amplification and Southern hybridization analysis. Using the yeast two-hybrid system, an interaction was observed between the full-length cytoplasmic domains of syntaxin 4 and synaptobrevin 2, a vesicle membrane SNARE previously shown by others to be associated with vesicles carrying the GLUT4 glucose transporter protein, but no interaction was seen with synaptobrevin 1. Flow cytometry of low-density microsomes isolated from fat cells was used to demonstrate the binding of syntaxin 4 to a subset of vesicles carrying GLUT4 protein; whereas SNAP-25 on its own bound poorly to these vesicles, the syntaxin 4-SNAP-25 complex gave a strong interaction.


Sponges as sentinels: patterns of spatial and intra-individual variation in trace metal concentration.
If sponges are to be effective biomonitors we require a better understanding of the spatial scales over which metals vary in these organisms. We determined how concentration of Cd, Zn, Cu, Pb, Hg and Se varied over four spatial scales for two common estuarine sponge species in the Sydney region. We examined variability with a fully nested sampling design; between coastal lakes, within coastal lakes, between sponges and within sponges. Calculation of variance components confirmed that 'within-sponge' variation in Cd, Zn, Cu, Pb and Se concentrations were low (1-14%) relative to the two largest spatial scales (49-98%) examined. In contrast, Hg concentrations exhibited marked variability 'between-sponges' and were below detection at one location. There was little evidence that sponge size was a good predictor of metal concentration. Taken together, these outcomes confirm that fragments of these sponges could be successfully transplanted and therefore show promise as biomonitors of metal contamination.


Mapping the interaction between the hemophore HasA and its outer membrane receptor HasR using CRINEPT-TROSY NMR spectroscopy.
The first step of heme acquisition by Gram-negative pathogenic bacteria through the so-called heme acquisition system, Has, requires delivery of the heme from the extracellular hemophore protein HasA to a specific outer membrane receptor, HasR. CRINEPT-TROSY NMR experiments in DPC micelles were here used to obtain information on the intermediate HasA-HasR complex in solution. A stable protein-protein adduct is detected both in the presence and in the absence of heme. Structural information on the complexed form of HasA is obtained from chemical shift mapping and statistical analysis of the spectral fingerprint of the protein NMR spectra obtained under different conditions. This approach shows the following: (i) only three different conformations are possible for HasA in solution: one for the isolated apoprotein, one for the isolated holoprotein, and one for the complexed protein, that is independent of the presence of the heme; (ii) the structure of the hemophore in the complex resembles the open conformation of the apoprotein; (iii) the surface contact area between HasA and HasR is independent of the presence of the heme, involving loop L1, loop L2, and the beta2-beta6 strands; (iv) upon complex formation the heme group is transferred from holoHasA to HasR.


Clinical geography: nursing practice and the (re)making of institutional space.
To present a geographical study that highlights the wide ranging spatial features of nursing agency. In turn, illustrate the further potential for geographical research to describe, support, challenge and guide clinical practice - particularly with regard to those 'everyday' activities and actions undertaken on a frequent basis. To provide a focus, and to anchor the study in existing clinical knowledge and debates, the role of nursing in the (re)making of institutional experiences and life is specifically explored. In-depth semi-structured interviews were conducted with 15 nurses working in Buckinghamshire and West London, UK. The following specialties were represented: acute care including emergency (n = 3), midwifery (n = 3), children's nursing (n = 2), elderly care (n = 1), rehabilitation (n = 2), mental health (n = 3) and palliative care (n = 1). Nurses claimed to actively manipulate, normalize and recreate clinical spaces as part of their everyday therapeutic practice. Specifically, the range of agency employed by them falls under the following six categories: adjusting social composition; introducing 'normal' activities; providing private spaces; seeking private spaces; explaining clinical spaces; spaces for personal escape and wellbeing. It is recommended that nurse leaders - including researchers, managers and clinical educators - explore geography as a source of social scientific evidence that sheds light on the complex nature of everyday professional practice. In this regard, some important disciplinary and structural issues are noted.


Interaction between artemether-lumefantrine and nevirapine-based antiretroviral therapy in HIV-1-infected patients.
Artemether-lumefantrine and nevirapine-based antiretroviral therapy (ART) are the most commonly recommended first-line treatments for malaria and HIV, respectively, in Africa. Artemether, lumefantrine, and nevirapine are metabolized by the cytochrome P450 3A4 enzyme system, which nevirapine induces, creating potential for important drug interactions. In a parallel-design pharmacokinetic study, concentration-time profiles were obtained in two groups of HIV-infected patients: ART-naïve patients and those stable on nevirapine-based therapy. Both groups received the recommended artemether-lumefantrine dose. Patients were admitted for intense pharmacokinetic sampling (0 to 72 h) with outpatient sampling until 21 days. Concentrations of lumefantrine, artemether, dihydroartemisinin, and nevirapine were determined by validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. The primary outcome was observed day 7 lumefantrine concentrations, as these are associated with therapeutic response in malaria. We enrolled 36 patients (32 females). Median (range) day 7 lumefantrine concentrations were 622 ng/ml (185 to 2,040 ng/ml) and 336 ng/ml (29 to 934 ng/ml) in the nevirapine and ART-naïve groups, respectively (P = 0.0002). The median artemether area under the plasma concentration-time curve from 0 to 8 h [AUC((0-8 h))] (P < 0.0001) and dihydroartemisinin AUC((60-68 h)) (P = 0.01) were lower in the nevirapine group. Combined artemether and dihydroartemisinin exposure decreased over time only in the nevirapine group (geometric mean ratio [GMR], 0.76 [95% confidence interval {CI}, 0.65 to 0.90]; P < 0.0001) and increased with the weight-adjusted artemether dose (GMR, 2.12 [95% CI, 1.31 to 3.45]; P = 0.002). Adverse events were similar between groups, with no difference in electrocardiographic Fridericia corrected QT and P-R intervals at the expected time of maximum lumefantrine concentration (T(max)). Nevirapine-based ART decreased artemether and dihydroartemisinin AUCs but unexpectedly increased lumefantrine exposure. The mechanism of the lumefantrine interaction remains to be elucidated. Studies investigating the interaction of nevirapine and artemether-lumefantrine in HIV-infected patients with malaria are urgently needed.


Posttranslational, site-directed photochemical fluorine editing of protein sidechains to probe residue oxidation state via 19F-nuclear magnetic resonance.
The fluorination of amino acid residues represents a near-isosteric alteration with the potential to report on biological pathways, yet the site-directed editing of carbon-hydrogen (C-H) bonds in complex biomolecules to carbon-fluorine (C-F) bonds is challenging, resulting in its limited exploitation. Here, we describe a protocol for the posttranslational and site-directed alteration of native γCH2 to γCF2 in protein sidechains. This alteration allows the installation of difluorinated sidechain analogs of proteinogenic amino acids, in both native and modified states. This chemical editing is robust, mild, fast and highly efficient, exploiting photochemical- and radical-mediated C-C bonds grafted onto easy-to-access cysteine-derived dehydroalanine-containing proteins as starting materials. The heteroaryl-sulfonyl reagent required for generating the key carbon-centered C• radicals that install the sidechain can be synthesized in two to six steps from commercially available precursors. This workflow allows the nonexpert to create fluorinated proteins within 24 h, starting from a corresponding purified cysteine-containing protein precursor, without the need for bespoke biological systems. As an example, we readily introduce three γCF2-containing methionines in all three progressive oxidation states (sulfide, sulfoxide and sulfone) as D-/L- forms into histone eH3.1 at site 4 (a relevant lysine to methionine oncomutation site), and each can be detected by 19F-nuclear magnetic resonance of the γCF2 group, as well as the two diastereomers of the sulfoxide, even when found in a complex protein mixture of all three. The site-directed editing of C-H→C-F enables the use of γCF2 as a highly sensitive, 'zero-size-zero-background' label in protein sidechains, which may be used to probe biological phenomena, protein structures and/or protein-ligand interactions by 19F-based detection methods.


Characterization of cholinesterases in the damselfish Sergeant major (Abudefduf saxatilis).
Cholinesterase (ChE) activity has been used for many years as a biomarker of exposure to organophosphate and carbamate pesticides. Recent studies have demonstrated that there could be biological factors that determine ChE type and levels; thus, juvenile Sergeant major (Abudefduf saxatilis) ChE enzymes were biochemically characterized. ChE enzymes found in the head and trunk were evaluated for their substrate preference and sensitivity to selective inhibitors. The use of the head and trunk was chosen as a strategy to reduce dissection time and to ensure sample uniformity between stations. The results indicated that there are two types of ChE enzymes in the head: acetylcholinesterase (AChE) and atypical butyrylcholinesterase (BChE) that exhibits intermediate characteristics of human AChE and BChE activities. Atypical BChE is predominantly found in the trunk. The results also indicated that the ChE activity found in A. saxatilis may be used as a biomarker in studies monitoring the Mexican Caribbean.


Two-view multibody structure-and-motion with outliers through model selection.
Multibody structure-and-motion (MSaM) is the problem to establish the multiple-view geometry of several views of a 3D scene taken at different times, where the scene consists of multiple rigid objects moving relative to each other. We examine the case of two views. The setting is the following: Given are a set of corresponding image points in two images, which originate from an unknown number of moving scene objects, each giving rise to a motion model. Furthermore, the measurement noise is unknown, and there are a number of gross errors, which are outliers to all models. The task is to find an optimal set of motion models for the measurements. It is solved through Monte-Carlo sampling, careful statistical analysis of the sampled set of motion models, and simultaneous selection of multiple motion models to best explain the measurements. The framework is not restricted to any particular model selection mechanism because it is developed from a Bayesian viewpoint: Different model selection criteria are seen as different priors for the set of moving objects, which allow one to bias the selection procedure for different purposes.


Thoughts about eating, weight and shape in anorexia nervosa and bulimia nervosa.
Concurrent verbalisation and a self-report questionnaire were used to investigate self-statements in patients with anorexia nervosa, patients with bulimia nervosa, two groups of dieters and non-dieting controls. Thoughts were collected while subjects performed three behavioural tasks, looking at themselves in a full-length mirror, weighing themselves and eating a chocolate covered mint. Both groups of patients had more negative thoughts related to eating, weight and shape than those in the three control groups. In addition, patients with anorexia nervosa showed a greater concern with eating while patients with bulimia nervosa showed a greater concern with weight and appearance. Differences were found between the patients and non-dieting controls using both methods but the self-report questionnaire was less sensitive than concurrent verbalisation to differences between the patients and dieters. Implications of the findings for cognitive-behavioural treatments of the two disorders are discussed.


Specific detection and real-time PCR quantification of potentially mycophagous bacteria belonging to the genus Collimonas in different soil ecosystems.
The bacterial genus Collimonas has the remarkable characteristic that it grows at the expense of living fungal hyphae under laboratory conditions. Here, we report the first field inventory of the occurrence and abundance of Collimonas in soils (n = 45) with naturally different fungal densities, which was performed in order to test the null hypothesis that there is a relationship between the presence of Collimonas and fungal biomass. Estimates of fungal densities were based on ergosterol measurements. Each soil was also characterized in terms of its physical and chemical properties and vegetation and management types. Culturable Collimonas was identified in plate-spread soil samples by its ability to clear colloidal chitin, in combination with a Collimonas-specific restriction fragment length polymorphism analysis of 16S rRNA PCR amplified from individual colonies. Using this approach, we found culturable collimonads only in (semi)natural grasslands. A real-time PCR assay for the specific quantification of Collimonas 16S rRNA in total soil DNA was developed. Collimonas was detectable in 80% of the soil samples, with densities up to 10(5) cells g(-1) (dry weight) soil. The numbers of Collimonas cells per gram of soil were consistently lowest in fungus-poor arable soils and, surprisingly, also in fungus-rich organic layers of forest soils. When all soils were included, no significant correlation was observed between the number of Collimonas cells and ergosterol-based soil fungal biomass. Based on this result, we rejected our null hypothesis, and possible explanations for this were addressed.


Using the protirelin test to distinguish mania from schizophrenia.
To explore the possible utility of the protirelin test in differentiating manic and schizophrenic patients, we gave a test dose of protirelin to 30 consecutive euthyroid inpatients who met Research Diagnostic Criteria for mania, 30 who met criteria for schizophrenia, undifferentiated subtype, and 20 normal volunteer controls. The mean maximal thyroid-stimulating hormone (TSH) response (delta TSH) to protirelin in the manic patients was lower than in the schizophrenic patients and in the controls. This mean difference was not attributable to differences in age, sex, baseline thyroid functioning, cortisol levels, or medication, but there was a considerable overlap of values in the patient groups. However, with a delta TSH less than or equal to 7.0 I microunits/mL to identify manic patients in the overall group, the sensitivity of the protirelin test was 60% and the specificity was 84%.


A neuraminidase from Streptococcus pneumoniae has the features of a surface protein.
A gene from Streptococcus pneumoniae (nanA), with features entirely consistent with a neuraminidase gene, has been sequenced. High levels of neuraminidase activity were obtained after cloning of this gene, without flanking sequences, into a high-expression vector. RNA hybridization studies have shown that the gene is transcribed by a virulent pneumococcus strain. The predicted molecular weight of the protein and certain amino acid sequences are typical of other neuraminidases. NanA contains the four copies of the sequence SXDXGXTW that is present in all the bacterial neuraminidases previously described. Kyte and Doolittle analysis showed that NanA is a hydrophilic protein with hydrophobic domains at the N terminus and the C terminus. A putative signal peptide was found in the N terminus of this protein, indicating that the protein is exported from the pneumococcus. The C terminus has the features of the anchor motif found in other surface proteins from gram-positive bacteria. Electron microscopy studies showed the presence of neuraminidase associated with the cell surface of the pneumococcus.


Bioprinting a Multifunctional Bioink to Engineer Clickable 3D Cellular Niches with Tunable Matrix Microenvironmental Cues.
The properties of the surrounding cell environment are major determinants of cell response in 3D. However, the ability to unravel how these cues dictate the biological function in bioprinted constructs is limited by the lack of extracellular matrix (ECM)-mimetic bioinks with fully controllable properties. In this study, a multifunctional bioink that uniquely combines the independent control over the biochemical and biophysical cues that regulate cell fate with the bioorthogonal nature of thiol-norbornene photoclick chemistry is designed for the extrusion bioprinting of bioinspired 3D cellular niches with tunable properties. The bioink rheology is controlled by ionic gelation, being dependent on both the type and content of divalent ions (calcium and barium), while the mechanical and biochemical properties of hydrogels are tailored via a post printing thiol-ene reaction. Bioprinted cell-adhesive and protease-degradable hydrogels modulate cell proliferation and ECM deposition in a matrix-stiffness dependent manner over 14 days of culture regardless of cell spreading, demonstrating the ability to probe the effect of matrix cues on cell response. This bioink can be used as a versatile platform where building blocks can be rationally combined for the bioprinting of functional cell- and tissue-specific constructs with controlled cellular behavior.


Complications of the 'treatment' of tonsillar infection by traditional healers in Nigeria.
Traditional healers in Nigeria attempt to treat tonsillar infections either by performing uvulectomy with a sickle knife or by digital rupture of the tonsil. These procedures result in various complications, including haemorrhage, septicaemia, cellulitis of the neck, peritonsillar and parapharyngeal abscess, upper airway obstruction, and pharyngo-laryngocele with pneumothorax. These complications are illustrated with case reports. The danger of encouraging traditional healers to practise unsupervised and unscientific medicine is stressed.


Changes in adipose tissue lipolysis gene expression and insulin sensitivity after weight loss.
Insulin resistance is a major pathophysiological link between obesity and its metabolic complications. Weight loss (WL) is an effective tool to prevent obesity-related diseases; however, the mechanisms of an improvement in insulin sensitivity (IS) after weight-reducing interventions are not completely understood. The aim of the present study was to analyze the relationships between IS and adipose tissue (AT) expression of the genes involved in the regulation of lipolysis in obese subjects after WL. Fifty-two obese subjects underwent weight-reducing dietary intervention program. The control group comprised 20 normal-weight subjects, examined at baseline only. Hyperinsulinemic-euglycemic clamp and s.c. AT biopsy with subsequent gene expression analysis were performed before and after the program. AT expression of genes encoding lipases (PNPLA2, LIPE and MGLL) and lipid-droplet proteins enhancing (ABHD5) and inhibiting lipolysis (PLIN1 and CIDEA) were decreased in obese individuals in comparison with normal-weight individuals. The group of 38 obese participants completed dietary intervention program and clamp studies, which resulted in a significant WL and an improvement in mean IS. However, in nine subjects from this group IS did not improve in response to WL. AT expression of PNPLA2, LIPE and PLIN1 increased only in the group without IS improvement. Excessive lipolysis may prevent an improvement in IS during WL. The change in AT PNPLA2 and LIPE expression was a negative predictor of the change in IS after WL.


Systematic review of the impact of breast-conserving surgery on cancer outcomes of multiple ipsilateral breast cancers.
The clinical effectiveness of treating ipsilateral multifocal (MF) and multicentric (MC) breast cancers using breast-conserving surgery (BCS) compared with the standard of mastectomy is uncertain. Inconsistencies relate to definitions, incidence, staging and intertumoral heterogeneity. The primary aim of this systematic review was to compare clinical outcomes after BCS versus mastectomy for MF and MC cancers, collectively defined as multiple ipsilateral breast cancers (MIBC). Comprehensive electronic searches were undertaken to identify complete papers published in English between May 1988 and July 2015, primarily comparing clinical outcomes of BCS and mastectomy for MIBC. All study designs were included, and studies were appraised critically using the Newcastle-Ottawa Scale. The characteristics and results of identified studies were summarized. Twenty-four retrospective studies were included in the review: 17 comparative studies and seven case series. They included 3537 women with MIBC undergoing BCS; breast cancers were defined as MF in 2677 women, MC in 292, and reported as MIBC in 568. Six studies evaluated MIBC treated by BCS or mastectomy, with locoregional recurrence (LRR) rates of 2-23 per cent after BCS at median follow-up of 59·5 (i.q.r. 56-81) months. BCS and mastectomy showed apparently equivalent rates of LRR (risk ratio 0·94, 95 per cent c.i. 0·65 to 1·36). Thirteen studies compared BCS in women with MIBC versus those with unifocal cancers, reporting LRR rates of 2-40 per cent after BCS at a median follow-up of 64 (i.q.r. 57-73) months. One high-quality study reported 10-year actuarial LRR rates of 5·5 per cent for BCS in 300 women versus 6·5 per cent for mastectomy among 887 women. The available studies were mainly of moderate quality, historical and underpowered, with limited follow-up and biased case selection favouring BCS rather than mastectomy for low-risk patients. The evidence was inconclusive, weakening support for the St Gallen consensus and supporting a future randomized trial.


Anti-cyclic citrullinated peptide antibody isotypes in rheumatoid arthritis: association with disease duration, rheumatoid factor production and the presence of shared epitope.
Anti-cyclic citrullinated peptide (anti-CCP) antibodies of IgG isotype are specific diagnostic markers of rheumatoid arthritis (RA). Recent evidence also points to their direct involvement in the pathophysiology. Little information is available, however, regarding the isotype distribution of anti-CCP antibodies and the characteristics of IgA and IgM anti-CCP. IgG, IgA and IgM anti-CCP2 and rheumatoid factor (RF) levels were measured in the sera of 119 RA patients and 118 controls, including patients with other rheumatic diseases and healthy subjects. We analyzed the diagnostic performance of IgA and IgM anti-CCP2 antibodies and their relationship with IgG anti-CCP2, RFs, disease duration and the presence of HLA-DRB1 shared epitope (SE) alleles. Patients with RA had significantly higher serum IgA and IgM anti-CCP2 antibody levels than healthy subjects and patients with other rheumatic diseases (p<0.0001). IgG, IgA and IgM anti-CCP2 antibodies were present in 74.8%, 52.9% and 44.5% of RA patients, and their diagnostic specificity was 95.8%, 95.8% and 91.6%, respectively. The presence of anti-CCP2 antibodies was significantly associated with SE alleles (p=0.03). The frequency of IgM anti-CCP2 positivity was lower in longstanding disease compared to early RA (p=0.03). IgA and IgM anti-CCP2 antibodies are present in RA patients, and they are similarly specific for RA as IgG anti-CCP2. The higher frequency of IgM anti-CCP2 antibodies in early RA suggests that they are mostly generated during the first phase of immune response; nonetheless, their production seems to be sustained in some patients. Further analysis of IgM and IgA anti-CCP2 antibodies may provide insights into the pathogenesis of RA.


Evaluating the Photoprotective Effects of Ochre on Human Skin by In Vivo SPF Assessment: Implications for Human Evolution, Adaptation and Dispersal.
Archaeological indicators of cognitively modern behaviour become increasingly prevalent during the African Middle Stone Age (MSA). Although the exploitation of ochre is viewed as a key feature of the emergence of modern human behaviour, the uses to which ochre and ochre-based mixtures were put remain ambiguous. Here we present the results of an experimental study exploring the efficacy of ochre as a topical photoprotective compound. This is achieved through the in vivo calculation of the sun protection factor (SPF) values of ochre samples obtained from Ovahimba women (Kunene Region, Northern Namibia) and the Palaeozoic Bokkeveld Group deposits of the Cape Supergroup (Western Cape Province, South Africa). We employ visible spectroscopy, energy-dispersive X-ray fluorescence (ED-XRF), X-ray diffraction (XRD) and granulometric analyses to characterise ochre samples. The capacity of ochre to inhibit the susceptibility of humans to the harmful effects of exposure to ultraviolet radiation (UVR) is confirmed and the mechanisms implicated in the efficacy of ochre as a sunscreen identified. It is posited that the habitual application of ochre may have represented a crucial innovation for MSA humans by limiting the adverse effects of ultraviolet exposure. This may have facilitated the colonisation of geographic regions largely unfavourable to the constitutive skin colour of newly arriving populations.


Evaluation of alginate hydrogel cytotoxicity on three-dimensional culture of type A spermatogonial stem cells.
The culture of spermatogonial cells for future transplantation, based on the specific biology of these cells is important and necessary. Recently, the use of scaffolds especially alginate for culturing stem cells has been the focus of many researchers. The aim of this study was to evaluate the cytotoxicity of alginate hydrogels to cultures of type A spermatogonial stem cells. Spermatogonial stem cells of 6day-old immature mice were isolated by surgery; thereafter, the cells were purified by MACS using antibodies against thy-1 and C-kit and cultured on a layer of laminin. After purification, spermatogonial stem cells were encapsulated in alginate hydrogels. After one month of encapsulation and culture in DMEM culture medium containing 10ng/ml GDNF, cells were removed from hydrogel and were examined for viability, cell morphology and structure, cytotoxicity and expression of apoptosis genes Fas, P53, Bax, Bcl2, Caspase3 by staining with trypan blue, scanning electron microscopy, LDH test, and Real time PCR, respectively. The encapsulation did not change the morphology and viability of spermatogonial stem cells. Investigations showed that spermatogonial stem cells preserve by the high viability (74.08%) and cytotoxicity of alginate hydrogel was estimated to be 5%. Expression of Fas gene increased in main group compared with the control group, and expression of Bax and P53 was reduced in main group compared with the control group. Expression of Bcl2 and Caspase3 genes did not show any significant difference between the main group and the control group. Considering the lack of cytotoxicity and antioxidant properties of alginate hydrogel scaffold and high viability of cells, this three-dimensional scaffold is applicable for culturing and encapsulation of spermatogonial stem cells.


Importance of resin ducts in reducing ponderosa pine mortality from bark beetle attack.
The relative importance of growth and defense to tree mortality during drought and bark beetle attacks is poorly understood. We addressed this issue by comparing growth and defense characteristics between 25 pairs of ponderosa pine (Pinus ponderosa) trees that survived and trees that died from drought-associated bark beetle attacks in forests of northern Arizona, USA. The three major findings of our research were: (1) xylem resin ducts in live trees were >10% larger (diameter), >25% denser (no. of resin ducts mm(-2)), and composed >50% more area per unit ring growth than dead trees; (2) measures of defense, such as resin duct production (no. of resin ducts year(-1)) and the proportion of xylem ring area to resin ducts, not growth, were the best model parameters of ponderosa pine mortality; and (3) most correlations between annual variation in growth and resin duct characteristics were positive suggesting that conditions conducive to growth also increase resin duct production. Our results suggest that trees that survive drought and subsequent bark beetle attacks invest more carbon in resin defense than trees that die, and that carbon allocation to resin ducts is a more important determinant of tree mortality than allocation to radial growth.


Non-allergic rhinitis: Position paper of the European Academy of Allergy and Clinical Immunology.
This EAACI position paper aims at providing a state-of-the-art overview on nonallergic rhinitis (NAR). A significant number of patients suffering from persistent rhinitis are defined as nonallergic noninfectious rhinitis (NANIR) patients, often denominated in short as having NAR. NAR is defined as a symptomatic inflammation of the nasal mucosa with the presence of a minimum of two nasal symptoms such as nasal obstruction, rhinorrhea, sneezing, and/or itchy nose, without clinical evidence of endonasal infection and without systemic signs of sensitization to inhalant allergens. Symptoms of NAR may have a wide range of severity and be either continuously present and/or induced by exposure to unspecific triggers, also called nasal hyperresponsiveness (NHR). NHR represents a clinical feature of both AR and NAR patients. NAR involves different subgroups: drug-induced rhinitis, (nonallergic) occupational rhinitis, hormonal rhinitis (including pregnancy rhinitis), gustatory rhinitis, senile rhinitis, and idiopathic rhinitis (IR). NAR should be distinguished from those rhinitis patients with an allergic reaction confined to the nasal mucosa, also called "entopy" or local allergic rhinitis (LAR). We here provide an overview of the current consensus on phenotypes of NAR, recommendations for diagnosis, a treatment algorithm, and defining the unmet needs in this neglected area of research.


Cyclopiazonic acid causes endothelium-dependent relaxation in rat aorta.
The effects of cyclopiazonic acid (CPA), a selective inhibitor of Ca(2+)-pump ATPase for endoplasmic reticulum (ER), on the contractility of rat aorta with and without intact endothelium were studied to investigate the possible involvement of endothelial ER Ca(2+)-pump in the release of endothelium-derived relaxing factor (EDRF), which is known to cause vascular relaxation or inhibition of phenylephrine (PE)-precontracted aorta. When added to the organ bath cumulatively, CPA concentration-dependently caused gradual development of contraction, which was much less in aortic rings with intact endothelium than in endothelium-denuded aortic rings. But CPA at low concentrations (1-3 mumol.L-1) induced vascular relaxation when added to PE (3 mumol.L-1)-precontracted aortic rings with intact endothelium, but not in denuded aortic rings. This relaxant effect of CPA is very similar to the effect of acetylcholine (ACh), which is well recognized to be mediated by the release of EDRF from the endothelium. NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, completely prevented the vascular relaxation induced by CPA or ACh and the inhibitory effect of L-NAME was partially reversed by L-arginine (L-Arg). Treatment of the aortic rings with nifedipine (Nif) 0.3 mumol.L-1 did not affect the relaxant effect of ACh or CPA on PE-induced contraction indicating that the Ca(2+)-entry to the endothelial cells as a result of receptor activation by ACh or ER Ca(2+)-pump inhibition by CPA was via channels other than L-type Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)


Physicians' ability to provide initial primary care to an HIV-infected patient.
Primary care physicians are providing care for an increasing number of persons infected with the human immunodeficiency virus (HIV). This study assesses the ability of primary care physicians to provide initial care for a patient with recently diagnosed HIV infection. A standardized patient (SP) was trained to portray an asymptomatic person with HIV infection seeking a primary care physician. Physicians took a medical history and counseled the SP; their performances were assessed by the SP and through a brief written examination. In addition, physicians distributed questionnaires to HIV-infected patients in their practices to assess actual performance. The study participants consisted of 121 primary care physicians. A minority of physicians recommended standard primary care screening tests and vaccinations, including viral hepatitis screening (35%), syphilis serologic testing (32%), and pneumococcal vaccination (23%). While most physicians (87%) indicated they would obtain CD4 cell counts, only 50% indicated they would start appropriate Pneumocystis carinii pneumonia prophylaxis. Although this patient presented documentation of a positive tuberculin skin test and no prior therapy for tuberculosis, only 53% of the physicians recommended prophylactic isoniazid. While 75% of the physicians asked this SP about condom use, number of sexual partners, or contact with previous sexual partners. Physicians with the most HIV experience were more likely to recommended P carinii pneumonia prophylaxis, coinfection screening, pneumococcal vaccination, and isoniazid prophylactic therapy. However, physicians' HIV experience was not associated with assessing this SP's risk of infecting others or with counseling regarding condom use. Questionnaires distributed to HIV-infected patients of these physicians generally confirmed these findings. Basic HIV preventive and primary care may not be adequately performed by many primary care physicians. Physicians' HIV experience was associated with better performance of HIV primary care tasks, but not with screening and counseling concerning the spread of HIV infection.


Effects of a knee ligament injury prevention exercise program on impact forces in women.
Previous research suggests high impact forces generated during landings contribute to noncontact anterior cruciate ligament (ACL) injuries. In women, neuromuscular differences appear to modify the ability to dissipate landing forces when compared to men. This study examined peak vertical impact forces (F(p)) and rate of force development (RFD) following a 9-week, low-intensity (simple jump-landing-jump tasks) and volume (number of foot contacts per workout) plyometric-based knee ligament injury prevention (KLIP) program. Female subjects were randomly assigned into control (n = 14) and treatment (n = 14) groups. Treatment subjects attended KLIP sessions twice a week for 9 weeks, and control subjects received no intervention. Ground reaction forces (F(p) and RFD) generated during a step-land protocol were assessed at study onset and termination. Significant reductions in F(p) (p = 0.0004) and RFD (p = 0.0205) were observed in the treatment group. Our results indicate that 9 weeks of KLIP training altered landing strategies in women to lower F(p) and RFD. These changes are considered conducive to a reduced risk of knee injury while landing.


Is REST required for ESC pluripotency?
The DNA-binding protein REST (also called NRSF) is a transcriptional repressor that targets many neuronal genes and is abundant in human and mouse pluripotent embryonic stem cells (ESCs). In a recent Letter to Nature, Singh et al. suggested that REST controls the self-renewal and pluripotency of ESCs, because they found that ESCs in which a single REST allele was disrupted (Fig. 1a, beta-geo-stop insertion) had reduced alkaline phosphatase activity and expressed lower levels of several pluripotency-associated genes. Here we show that partial or complete loss of functional REST protein does not abrogate ESC potential as reflected by marker gene expression. These data are consistent with earlier reports, and argue that REST is not required for maintaining ESC pluripotency.


Disparity of Cytochrome Utilization in Anodic and Cathodic Extracellular Electron Transfer Pathways of Geobacter sulfurreducens Biofilms.
Extracellular electron transfer (EET) in microorganisms is prevalent in nature and has been utilized in functional bioelectrochemical systems. EET of Geobacter sulfurreducens has been extensively studied and has been revealed to be facilitated through c-type cytochromes, which mediate charge between the electrode and G. sulfurreducens in anodic mode. However, the EET pathway of cathodic conversion of fumarate to succinate is still under debate. Here, we apply a variety of analytical methods, including electrochemistry, UV-vis absorption and resonance Raman spectroscopy, quartz crystal microbalance with dissipation, and electron microscopy, to understand the involvement of cytochromes and other possible electron-mediating species in the switching between anodic and cathodic reaction modes. By switching the applied bias for a G. sulfurreducens biofilm coupled to investigating the quantity and function of cytochromes, as well as the emergence of Fe-containing particles on the cell membrane, we provide evidence of a diminished role of cytochromes in cathodic EET. This work sheds light on the mechanisms of G. sulfurreducens biofilm growth and suggests the possible existence of a nonheme, iron-involving EET process in cathodic mode.


T cell receptor-mediated signaling events in CD4+CD8+ thymocytes undergoing thymic selection: requirement of calcineurin activation for thymic positive selection but not negative selection.
The goal of this study was to identify the differences of intracellular signals between the processes of thymic positive and negative selection. The activation of calcineurin, a calcium- and calmodulin-dependent phosphatase, is known to be an essential event in T cell activation via the T cell receptor (TCR). The effect of FK506, an inhibitor of calcineurin activation, on positive and negative selection in CD4+CD8+ double positive (DP) thymocytes was examined in normal mice and in a TCR transgenic mouse model. In vivo FK506 treatment blocked the generation of mature TCRhighCD4+CD8- and TCRhighCD4-CD8+ thymocytes, and the induction of CD69 expression on DP thymocytes. In addition, the shutdown of recombination activating gene 1 (RAG-1) transcription and the downregulation of CD4 and CD8 expression were inhibited by FK506 treatment suggesting that the activation of calcineurin is required for the first step (or the very early intracellular signaling events) of TCR-mediated positive selection of DP thymocytes. In contrast, FK506-sensitive calcineurin activation did not appear to be required for negative selection based on the observations that negative selection of TCR alpha beta T cells in the H-2b male thymus (a negative selecting environment) was not inhibited by in vivo treatment with FK506 and that there was no rescue of the endogenous superantigen-mediated clonal deletion of V beta 6 and V beta 11 thymocytes in FK506-treated CBA/J mice. DNA fragmentation induced by TCR activation of DP thymocytes in vitro was not affected by FK506. In addition, different effects of FK506 from Cyclosporin A on the T cell development in the thymus were demonstrated. The results of this study suggest that different signaling pathways work in positive and negative selection and that there is a differential dependence on calcineurin activation in the selection processes.


Influenza vaccination among healthcare workers in Italy.
Influenza vaccination is a fundamental tool for the prevention of influenza in healthcare settings and its administration to healthcare workers (HCWs) is recommended in more than 40 countries including United States of America and many countries of the European Union. Despite these recommendations, the compliance of HCWs to influenza vaccination is largely inadequate in Italy. Since 2005/06 season, a comprehensive multifaceted intervention project aimed at increasing the seasonal influenza vaccination coverage rates among HCWs was performed at the IRCCS AOU San Martino IST teaching hospital in Genoa, Italy, the regional tertiary adult acute-care reference center with a 1300 bed capacity. Despite almost a decade of efforts, the vaccination coverage rates registered at our hospital steadily remain unsatisfactory and very distant by the minimum objective of 75% defined by the Italian Ministry of Health. During the last influenza season (2013/14), vaccination coverage rates by occupation type resulted 30% among physicians, 11% among nurses and 9% among other clinical personnel. Further efforts are necessary to prevent the transmission of influenza to patient and novel strategies need to be identified and implemented in order to increase the compliance of HCWs, particularly nurses, with the seasonal influenza vaccination.


Virtual education in neurosurgery during the COVID-19 pandemic.
Neurosurgical education in the US has changed significantly as a consequence of the novel coronavirus (COVID-19) pandemic. Institutional social distancing requirements have resulted in many neurosurgical programs utilizing video conferencing for educational activities. However, it is unclear how or if these practices should continue after the pandemic. The objective of this study was to characterize virtual education in neurosurgery and understand how it should be utilized after COVID-19. A 24-question, 3-part online survey was administered anonymously to all 117 US neurosurgical residency programs from May 15, 2020, to June 15, 2020. Questions pertained to the current use of virtual conferencing, preferences over traditional conferences, and future inclinations. The Likert scale (1 = strongly disagree, 3 = neutral, 5 = strongly agree) was used. Comparisons were calculated using the Mann-Whitney U-test. Statistical significance was set at 0.05. One-hundred eight responses were recorded. Overall, 38 respondents (35.2%) were attendings and 70 (64.8%) were trainees. Forty-one respondents (38.0%) indicated attending 5-6 conferences per week and 70 (64.8%) attend national virtual conferences. When considering different conference types, there was no overall preference (scores < 3) for virtual conferences over traditional conferences. In regard to future use, respondents strongly agreed that they would continue the practice at some capacity after the pandemic (median score 5). Overall, respondents agreed that virtual conferences would partially replace traditional conferences (median score 4), whereas they strongly disagreed with the complete replacement of traditional conferences (median score 1). The most common choices for the partial replacement of tradition conferences were case conferences (59/108, 55%) and board preparation (64/108, 59%). Lastly, there was a significant difference in scores for continued use of virtual conferencing in those who attend nationally sponsored conferences (median score 5, n = 70) and those who do not (median score 4, n = 38; U = 1762.50, z = 2.97, r = 0.29, p = 0.003). Virtual conferences will likely remain an integral part of neurosurgical education after the COVID-19 pandemic has abated. Across the country, residents and faculty report a preference for continued use of virtual conferencing, especially virtual case conferences and board preparation. Some traditional conferences may even be replaced with virtual conferences, in particular those that are more didactic. Furthermore, nationally sponsored virtual conferences have a positive effect on the preferences for continued use of virtual conferences.


Development of a dual-path system for band-to-band registration of an acousto-optic tunable filter-based imaging spectrometer.
Accurate band-to-band registration is crucial when an acousto-optic tunable filter (AOTF)-based imaging spectrometer is used to acquire spectral data on an unstable platform. However, it is difficult to registrate hyperspectral images accurately using traditional image registration algorithms, which will seriously affect the accurate acquisition of spectra and reduce the classification accuracy. Here we demonstrate an optical configuration that acquires the diffracted and undiffracted beam of the AOTF simultaneously. These two beams have identical geometrical optical characteristic and suffer from the same image motion caused by the platform jitter. Compared with the diffracted beam, however, the intensity of the undiffracted beam is changeless when tuning the AOTF. Therefore, the use of undiffracted beams allows accurate measurement of image motion in different bands. The experiment shows that this technique greatly improves the registration accuracy of spectral images with poor correlation and low SNR.


Ischaemic preconditioning reduces troponin T release in patients undergoing coronary artery bypass surgery.
To investigate whether ischaemic preconditioning could reduce myocardial injury, as manifest by troponin T release, in patients undergoing elective coronary artery bypass surgery. Randomised controlled trial. Cardiothoracic unit of a tertiary care centre. Patients with three vessel coronary artery disease and stable angina admitted for first time elective coronary artery bypass surgery were invited to take part in the study; 33 patients were randomised into control or preconditioning groups. Patients in the preconditioning group were exposed to two additional three minute periods of myocardial ischaemia at the beginning of the revascularisation operation, before the ischaemic period used for the first coronary artery bypass graft distal anastomosis. Serum troponin T concentration at 72 hours after cardiopulmonary bypass. The troponin T assays were performed by blinded observers at a different hospital. All patients had undetectable serum troponin T (< 0.1 microgram/l) before cardiopulmonary bypass, and troponin T was raised postoperatively in all patients. At 72 hours, serum troponin T was lower (P = 0.05) in the preconditioned group (median 0.3 microgram/l) than in the control group (median 1.4 micrograms/l). The direct application of a preconditioning stimulus in clinical practice has been shown, for the first time, to protect patients against irreversible myocyte injury.


Lower-half facial migraine: a report of 11 cases.
Vascular pain of the face constitutes a variant of pain of the head, and includes migraine, cluster headache, paroxysmal hemicrania, and a facial variant of the so-called lower-half migraine. Lower-half facial migraine is a condition difficult to classify; according to the international classifications it could not be found as an individual entity. The objective of the present study is to determine the difficulties we encountered in diagnosis, the ineffective treatments provided, and the pharmacologic treatment effect. A study is made of 11 cases of lower-half facial migraine, corresponding to 10 women and 1 man (mean age, 35 years), commenting on the clinical characteristics of the disorder and its treatment options. The location of the pain often mimics dental pain, and can lead to a mistaken diagnosis and to the application of inappropriate therapeutic measures. Forty-five percent of the patients had a history of endodontic treatment before the development of pain in the initially affected quadrant. Once the pain had developed, extractions were carried out in 36% of cases in an unsuccessful attempt to secure symptom relief. Our pharmacologic treatment consisted of ergotamine in 9 cases and the remaining 2 patients received indomethacin. Nine patients (82%) improved as a result of treatment, with an important reduction in the frequency of the pain episodes and intensity of pain. One patient failed to respond to ergotamine, while another patient failed to improve with indomethacin. Both were prescribed only minor analgesics. The treatment of migraine occurring in the face is no different than that provided for pain occurring in the head.


Pain assessment and management in different wards of a tertiary care hospital.
To assess the pain management by medical team, emergency room (ER) team and Acute Pain team in a tertiary care hospital. The cross-sectional study was done in Medical Ward, Surgical Ward and Emergency Room of Aga Khan University, Karachi, in March-April 2010. The assigned research medical officer visited the three locations every day and selected patients by way of convenient sampling. The study comprised 75 patients; 25 each in three groups. Information was collected on patient's demographics, general characteristics, type of drugs and modalities used. Specific queries about pain were sorted out like adequacy of pain assessment done by primary physician, pain intensity, any intervention done and pain relief post-intervention. SPSS version 17, analysis of variance and Chi square test were used for statistical purpose. The mean current pain score on the visual analogue score (VAS) was lowest in the Surgical Ward which was being managed by the Acute Pain Management Service (APMS) team followed by the Medical Ward and then Emergency Rooms. The difference was found to be statistically significant. The mean of worst pain score was also the lowest in the Surgical Ward. There was significant difference between wards in terms of the use of pain medications. Proper documentation for pain was done for all patients in the Surgical Ward, followed by the Emergency Room and then the Medical Ward. Better pain assessment, re-assessment, documentation and patient satisfaction were observed in the Surgical Ward compared to the other two locations of the study.


MiR-375 is Essential for Human Spinal Motor Neuron Development and May Be Involved in Motor Neuron Degeneration.
The transcription factor REST is a key suppressor of neuronal genes in non-neuronal tissues. REST has been shown to suppress proneuronal microRNAs in neural progenitors indicating that REST-mediated neurogenic suppression may act in part via microRNAs. We used neural differentiation of Rest-null mouse ESC to identify dozens of microRNAs regulated by REST during neural development. One of the identified microRNAs, miR-375, was upregulated during human spinal motor neuron development. We found that miR-375 facilitates spinal motor neurogenesis by targeting the cyclin kinase CCND2 and the transcription factor PAX6. Additionally, miR-375 inhibits the tumor suppressor p53 and protects neurons from apoptosis in response to DNA damage. Interestingly, motor neurons derived from a spinal muscular atrophy patient displayed depressed miR-375 expression and elevated p53 protein levels. Importantly, SMA motor neurons were significantly more susceptible to DNA damage induced apoptosis suggesting that miR-375 may play a protective role in motor neurons.


Minimal Access Posterior Approach for Extrapleural Thoracic Sympathectomy: A Cadaveric Study and Cases.
Operatively, video-assisted thoracoscopic sympathectomy (VATS) involves pleural entry and poses risk in small children and patients with pulmonary disease. A conventional posterior sympathectomy is more invasive than VATS. We investigated a cadaveric feasibility study of a minimal access posterior approach for endoscopic extrapleural sympathectomy and discuss this minimal approach in children with cardiac sympathectomy. A posterior endoscopic extrapleural approach for thoracic sympathectomy was performed using lightly embalmed cadavers; surgical corridor depth, width, and associated pleural violation were recorded. Two pediatric cases undergoing secondary prevention for breakthrough cardiac dysrhythmias using this approach are discussed: case 1, a 9-year-old girl with refractory long QT syndrome; and case 2, a 13-year-old boy with hypertrophic cardiomyopathy. The cadaveric study supported 100% identification of a craniocaudal-oriented sympathetic chain using an 18-mm tubular retractor, and a 10% pleural violation rate. There were no clinically significant pneumothoracies in either proof of concept cases. Minimal access posterior extrapleural sympathectomy is feasible to expose the sympathetic chain in the thoracic region with good visualization using either endoscopic or microscopic magnification. Single-position bilateral thoracic sympathectomy can be performed in pediatric patients with life-threatening ventricular arrhythmias. Based on the cadaveric study and the 2 preliminary cases, we believe that a posterior minimal access approach allows safe and effective access to the thoracic sympathetic chain for causes requiring sympathectomy using single positioning, with minimal risk of pneumothorax or Horner syndrome.


Search for long-lived charged massive particles with the D0 detector.
We search for long-lived charged massive particles using 1.1 fb;{-1} of data collected by the D0 detector at the Fermilab Tevatron pp[over ] Collider. Time-of-flight information is used to search for pair produced long-lived tau sleptons, gauginolike charginos, and Higgsino-like charginos. We find no evidence of a signal and set 95% C.L. cross section upper limits for staus, which vary from 0.31 to 0.04 pb for stau masses between 60 and 300 GeV. We also set lower mass limits of 206 GeV (171 GeV) for pair produced charged gauginos (Higgsinos).


Cerebral glucose and lactate consumption during cerebral activation by physical activity in humans.
At rest, the brain takes up oxygen and carbohydrate at an ~6:1 ratio. Exercise increases systemic lactate availability reducing this to as little as 1.7:1 despite a ~20% increase in cerebral metabolic rate for oxygen (CMRo₂), thus indicating a disproportionate increase of carbohydrate metabolism. Underlining mechanisms and metabolic fate for the augmented lactate uptake are unknown. This meta-analysis examines whether adrenergic activation explains the increased lactate uptake, cerebral lactate release following cerebral activation compensates for the extra carbohydrate uptake during exercise, and cerebral lactate uptake spares glucose as fuel. Ten studies (n=96) measuring arteriovenous differences for lactate, glucose, and oxygen and cerebral blood flow were included. Cerebral lactate uptake increased during brain activation by whole-body exercise compared to the resting state. Unlike glucose, lactate uptake is proportional to its arterial concentration but is unaffected by sympathetic activity. Following exercise, significant cerebral lactate released as arterial lactate levels decreased, which may balance the surplus lactate uptake in the brain during physical activity in the long term. Finally, cerebral glucose uptake was reduced by ~25% in relation to CMRo₂ when cerebral lactate uptake increased, suggesting, in part, preferential lactate consumption during activation. This meta-analysis favors the notion that cerebral lactate uptake is mainly passively governed by its availability, but when lactate is available, lactate supplements glucose and supports an increase in cerebral energy metabolism in an activity-dependent manner.


A battery-less and wireless wearable sensor system for identifying bed and chair exits in a pilot trial in hospitalized older people.
Falls in hospitals are common, therefore strategies to minimize the impact of these events in older patients and needs to be examined. In this pilot study, we investigate a movement monitoring sensor system for identifying bed and chair exits using a wireless wearable sensor worn by hospitalized older patients. We developed a movement monitoring sensor system that recognizes bed and chair exits. The system consists of a machine learning based activity classifier and a bed and chair exit recognition process based on an activity score function. Twenty-six patients, aged 71 to 93 years old, hospitalized in the Geriatric Evaluation and Management Unit participated in the supervised trials. They wore over their attire a battery-less, lightweight and wireless sensor and performed scripted activities such as getting off the bed and chair. We investigated the system performance in recognizing bed and chair exits in hospital rooms where RFID antennas and readers were in place. The system's acceptability was measured using two surveys with 0-10 likert scales. The first survey measured the change in user perception of the system before and after a trial; the second survey, conducted only at the end of each trial, measured user acceptance of the system based on a multifactor sensor acceptance model. The performance of the system indicated an overall recall of 81.4%, precision of 66.8% and F-score of 72.4% for joint bed and chair exit recognition. Patients demonstrated improved perception of the system after use with overall score change from 7.8 to 9.0 and high acceptance of the system with score ≥ 6.7 for all acceptance factors. The present pilot study suggests the use of wireless wearable sensors is feasible for detecting bed and chair exits in a hospital environment.


Colorectal cancers show distinct mutation spectra in members of the canonical WNT signaling pathway according to their anatomical location and type of genetic instability.
It is unclear whether the mutation spectra in WNT genes vary among distinct types of colorectal tumors. We have analyzed mutations in specific WNT genes in a cohort of 52 colorectal tumors and performed a meta-analysis of previous studies. Notably, significant differences were found among the mutation spectra. We have previously shown that in familial adenomatous polyposis, APC somatic mutations are selected to provide the "just-right" level of WNT signaling for tumor formation. Here, we found that APC mutations encompassing at least two beta-catenin down-regulating motifs (20 a.a. repeats) are significantly more frequent in microsatellite unstable (MSI-H) than in microsatellite stable (MSS) tumors where truncations retaining less than two repeats are more frequent (P = 0.0009). Moreover, in cases where both APC hits are detected, selection for mutations retaining a cumulative number of two 20 a.a. repeats became apparent in MSI-H tumors (P = 0.001). This type of mutations were also more frequent in proximal versus distal colonic tumors, regardless of MSI status (P = 0.0008). Among MSI-H tumors, CTNNB1 mutations were significantly more frequent in HNPCC than in sporadic lesions (28% versus 6%, P < 10-6) and were preferentially detected in the proximal colon, independently of MSI status (P = 0.017). In conclusion, the observed spectra of WNT gene mutations in colorectal tumors are likely the result from selection of specific levels of beta-catenin signaling, optimal for tumor formation in the context of specific anatomical locations and forms of genetic instability. We suggest that this may underlie the preferential location of MMR deficient tumors in the proximal colon.