Polarized fluorescence depletion reports orientation distribution and rotational dynamics of muscle cross-bridges.
The method of polarized fluorescence depletion (PFD) has been applied to enhance the resolution of orientational distributions and dynamics obtained from fluorescence polarization (FP) experiments on ordered systems, particularly in muscle fibers. Previous FP data from single fluorescent probes were limited to the 2(nd)- and 4(th)-rank order parameters, <P(2)(cos beta)> and <P(4)(cos beta)>, of the probe angular distribution (beta) relative to the fiber axis and <P(2d)>, a coefficient describing the extent of rapid probe motions. We applied intense 12-micros polarized photoselection pulses to transiently populate the triplet state of rhodamine probes and measured the polarization of the ground-state depletion using a weak interrogation beam. PFD provides dynamic information describing the extent of motions on the time scale between the fluorescence lifetime (e.g., 4 ns) and the duration of the photoselection pulse and it potentially supplies information about the probe angular distribution corresponding to order parameters above rank 4. Gizzard myosin regulatory light chain (RLC) was labeled with the 6-isomer of iodoacetamidotetramethylrhodamine and exchanged into rabbit psoas muscle fibers. In active contraction, dynamic motions of the RLC on the PFD time scale were intermediate between those observed in relaxation and rigor. The results indicate that previously observed disorder of the light chain region in contraction can be ascribed principally to dynamic motions on the microsecond time scale.


Age decreases endothelial progenitor cell recruitment through decreases in hypoxia-inducible factor 1alpha stabilization during ischemia.
Advanced age is known to impair neovascularization. Because endothelial progenitor cells (EPCs) participate in this process, we examined the effects of aging on EPC recruitment and vascular incorporation. Murine neovascularization was examined by use of an ischemic flap model, which demonstrated aged mice (19 to 24 months) had decreased EPC mobilization (percent mobilized 1.4+/-0.2% versus 0.4+/-0.1%, P<0.005) that resulted in impaired gross tissue survival compared with young mice (2 to 6 months). This decrease correlated with diminished tissue perfusion (P<0.005) and decreased CD31+ vascular density (P<0.005). Gender-mismatched bone marrow transplantation demonstrated significantly fewer chimeric vessels in aged mice (P<0.05), which confirmed a deficit in bone marrow-mediated vasculogenesis. Age had no effect on total EPC number in mice or humans. Reciprocal bone marrow transplantations confirmed that impaired neovascularization resulted from defects in the response of aged tissue to hypoxia and not from intrinsic defects in EPC function. We demonstrate that aging decreased hypoxia-inducible factor 1alpha stabilization in ischemic tissues because of increased prolyl hydroxylase-mediated hydroxylation (P<0.05) and proteasomal degradation. This resulted in a diminished hypoxia response, including decreased stromal cell-derived factor 1 (P<0.005) and vascular endothelial growth factor (P<0.0004). This effect can be reversed with the iron chelator deferoxamine, which results in hypoxia-inducible factor 1alpha stabilization and increased tissue survival. Aging impairs EPC trafficking to sites of ischemia through a failure of aged tissues to normally activate the hypoxia-inducible factor 1alpha-mediated hypoxia response.


Follicular development and oocyte maturation in hypogonadotrophic women employing recombinant follicle-stimulating hormone: the role of oestradiol.
Both luteinizing hormone (LH) and follicle-stimulating hormone (FSH) are required for follicle development and oestrogen production. Moreover, under normal conditions a close association between dominant follicle size and serum and intrafollicular oestradiol levels is observed. With the recent availability of human recombinant FSH (recFSH), it was possible for the first time to study effects of FSH alone, in the complete absence of endogenous or exogenous LH, on ovarian function. Recent studies applying recFSH in hypogonadotrophic women have shown convincingly that normal growth of follicles up to the preovulatory stage occurs despite extremely low oestradiol levels, in keeping with previous observations using exogenous gonadotrophins in women incapable of synthesizing oestradiol due to steroid enzyme abnormalities. Insufficient data are presently available in humans to conclude whether or not oocyte quality is compromised under these circumstances. It should, however, be realized that sufficient oestradiol levels are required for fertilization in vivo. Therefore LH, or human chorionic gonadotrophin (HCG), should be added to stimulation protocols in hypogonadotrophic individuals. These observations may also be relevant for monitoring of ovarian response during recFSH therapy, especially when combined with gonadotrophin-releasing hormone agonists for ovarian hyperstimulation for in-vitro fertilization.


Integrated genome-wide analysis of transcription factor occupancy, RNA polymerase II binding and steady-state RNA levels identify differentially regulated functional gene classes.
Routine methods for assaying steady-state mRNA levels such as RNA-seq and micro-arrays are commonly used as readouts to study the role of transcription factors (TFs) in gene expression regulation. However, cellular RNA levels do not solely depend on activity of TFs and subsequent transcription by RNA polymerase II (Pol II), but are also affected by RNA turnover rate. Here, we demonstrate that integrated analysis of genome-wide TF occupancy, Pol II binding and steady-state RNA levels provide important insights in gene regulatory mechanisms. Pol II occupancy, as detected by Pol II ChIP-seq, was found to correlate better with TF occupancy compared to steady-state RNA levels and is thus a more precise readout for the primary transcriptional mechanisms that are triggered by signal transduction. Furthermore, analysis of differential Pol II occupancy and RNA-seq levels identified genes with high Pol II occupancy and relatively low RNA levels and vice versa. These categories are strongly enriched for genes from different functional classes. Our results demonstrate a complementary value in Pol II chip-seq and RNA-seq approaches for better understanding of gene expression regulation.


Telomerase activity and differential expression of telomerase genes and c-myc in chicken cells in vitro.
This study examined telomerase activity and gene expression profiles for three genes in Gallus gallus domesticus: telomerase reverse transcriptase (chTERT), telomerase RNA (chTR), and c-myc. Expression of these genes was studied in chicken embryonic stem (chES) cells, chicken embryo fibroblasts (CEFs), and DT40 cells using quantitative real-time polymerase chain reaction. Our results establish that, relative to transcription levels in telomerase-negative CEFs, chTERT and chTR are up-regulated in telomerase-positive chES cells. Transcription levels of chTERT, chTR, and c-myc are dramatically up-regulated in telomerase-positive DT40 cells, relative to CEFs and chES cells. These results are consistent with a model in which telomerase activity is up-regulated in proliferating embryonic stem cells requiring stable telomeres to endure multiple rounds of cell division; down-regulated in differentiated, lifespan-limited cells; and dramatically up-regulated in immortalized, transformed cells for which uncontrolled proliferation is correlated with c-myc dysregulation and telomerase activity.


Legionella contamination of dental-unit waters.
Water samples collected from 28 dental facilities in six U.S. states were examined for the presence of Legionella pneumophila and other Legionella spp. by the PCR-gene probe, fluorescent-antibody microscopic, and viable-plate-count detection methods. The PCR and fluorescent-antibody detection methods, which detect both viable and viable nonculturable Legionella spp., gave higher counts and rates of detection than the plate count method. By the PCR-gene probe detection method, Legionella spp. were detected in 68% of the dental-unit water samples and L. pneumophila was detected in 8%. Concentrations of Legionella spp. in dental-unit water reached 1,000 organisms per ml or more in 36% of the samples, and 19% of the samples were in the category of 10,000/ml or above. L. pneumophila, when present in dental-unit water, never reached concentrations of 1,000/ml or more. Microscopic examination with fluorescent-antibody staining indicated that the contamination was in the dental-unit water lines rather than in the handpieces. Legionella spp. were present in 61% of potable water samples collected for comparative analysis from domestic and institutional faucets and drinking fountains; this percentage was not significantly different from the rate of detection of Legionella spp. in dental-unit water. However, in only 4% of the potable water samples did Legionella spp. reach concentrations of 1,000 organisms per ml, and none was in the 10,000 organisms-per-ml category, and so health-threatening levels of Legionella spp. in potable water were significantly lower than in dental-unit water. L. pneumophila was found in 2% of the potable water samples, but only at the lowest detectable level.(ABSTRACT TRUNCATED AT 250 WORDS)


The cytokineplast: purified, stable, and functional motile machinery from human blood polymorphonuclear leukocytes.
We examined the formation of motile, chemotactically active, anucleate fragments from human blood polymorphonuclear leukocytes (PMN, granulocytes), induced by the brief application of heat. These granule-poor fragments are former protopods (leading fronts, lamellipodia) that become uncoupled from the main body of the cell and leave it, at first with a connecting filament that breaks and seals itself. The usual random orientation of such filaments can be controlled by preorientation of cells in a gradient of the chemotactic peptide, N-formylmethionylleucylphenylalanine (F-Met-Leu-Phe) (2x10(-9) M- 1x10(-8)). Cytochalsin B, 2.5-5 mug/ml, prevents fragment formation; colchicine, 10(-5) M, does not. In scanning electron micrographs, fragments are ruffled and the cell body rounded up and rather smooth. In transmission electron micrographs, fragments contain microfilaments but lack centrioles and microtubules. Like intact cells, both bound and free fragments can respond chemotactically to an erythrocyte destroyed by laser microirradiation (necrotaxis); the free, anucleate fragments may do so repeatedly, even after having been held overnight at ambient temperatures. We propse the name cytokineplast for the result of this self-purification of motile apparatus. The exodus of the motile machinery from the granulocyte requires anchoring of the bulk of the cell to glass and uncoupling, which may involve heat-induced dysfunction of the centrosome. In ultrastructural studies of the centrosomal region after heat, centriolar structure remains intact, but pericentriolar osmiophilic material appears condensed, and microtubules are sparse. These changes are found in all three blood cell types examined: PMN, eosinophil, and monocyte. Of these, the first two make fragments under our conditions; the more sluggish monocyte does not. Uncoupling is further linked to centrosomal dysfunction by the observation that colchicines-treated granulocytes (10(-5)M, to destroy the centrosome's efferent arm) make fragments after less heat than controls. If motive force and orientation are specified mainly from the organelle-excluding leading front, then endoplasmic streaming in PMN is a catch-up phenomenon, and microtubules do not provide the vector of locomotion but rather stabilize and orient the "baggage" (nucleus, granuloplasm)-i.e., they prevent fishtailing. Moreover, constraints emanating from the centrosome may now be extended to include, maintenance of the motile machinery as an integral part of the cell.


Physical activity at 36 years: patterns and childhood predictors in a longitudinal study.
The aim was to describe the sex and socioeconomic differences in patterns of physical activity at work and in leisure time of men and women aged 36 years, and to investigate factors in childhood and adolescence which predict high rates of participation in sports and recreational activities in later life. Data collected in childhood, adolescence, and at 36 years on members of a national prospective birth cohort study were used. The population sample was resident in England, Scotland, and Wales. A stratified sample of about 3500 men and women was studied regularly from birth until 43 years. More men than women reported high rates of sports and recreational activities, gardening, and do-it-yourself. In contrast women reported higher rates of bicycling and walking. Higher levels of education were associated with frequent participation in sports. Individuals often engaged in one type of activity without necessarily engaging in other types. Those who were most active in sport had been above average at sports in school, more outgoing socially in adolescence, had fewer health problems in childhood, were better educated, and had more mothers with a secondary education than those who were less active. Studies that examine the relationship between physical activity and chronic disease should consider a broad range of pursuits rather than extrapolating from only one area of physical activity, and in their explanations should take account of the possible role of childhood characteristics. The findings suggest the importance of developing skills and habits in childhood as well as of encouraging healthier exercise habits in adults who may have had few opportunities or low motivation previously.


Relationships Between Bioactive Compound Content and the Antiplatelet and Antioxidant Activities of Six Allium Vegetable Species.
Allium sp. vegetables are widely consumed for their characteristic flavour. Additionally, their consumption may provide protection against cardiovascular disease due to their antiplatelet and antioxidant activities. Although antiplatelet and antioxidant activities in Allium sp. are generally recognised, comparative studies of antiplatelet and antioxidant potency among the main Allium vegetable species are lacking. Also, the relationship between organosulfur and phenolic compounds and these biological activities has not been well established. In this study, the in vitro antiplatelet and antioxidant activities of the most widely consumed Allium species are characterised and compared. The species total organosulfur and phenolic content, and the HPLC profiles of 11 phenolic compounds were characterised and used to investigate the relationship between these compounds and antiplatelet and antioxidant activities. Furthermore, antiplatelet activities in chives and shallot have been characterised for the first time. Our results revealed that the strongest antiplatelet agents were garlic and shallot, whereas chives had the highest antioxidant activity. Leek and bunching onion had the weakest both biological activities. Significantly positive correlations were found between the in vitro antiplatelet activity and total organosulfur (R=0.74) and phenolic (TP) content (R=0.73), as well as between the antioxidant activity and TP (R=0.91) and total organosulfur content (R=0.67). Six individual phenolic compounds were associated with the antioxidant activity, with catechin, epigallocatechin and epicatechin gallate having the strongest correlation values (R>0.80). Overall, our results suggest that both organosulfur and phenolic compounds contribute similarly to Allium antiplatelet activity, whereas phenolics, as a whole, are largely responsible for antioxidant activity, with broad variation observed among the contributions of individual phenolic compounds.


The role of glucosamine sulfate and chondroitin sulfates in the treatment of degenerative joint disease.
Successful treatment of osteoarthritis must effectively control pain, and should slow down or reverse progression of the disease. Biochemical and pharmacological data combined with animal and human studies demonstrate glucosamine sulfate is capable of satisfying these criteria. Glucosamine sulfate's primary biological role in halting or reversing joint degeneration appears to be directly due to its ability to act as an essential substrate for, and to stimulate the biosynthesis of, the glycosaminoglycans and the hyaluronic acid backbone needed for the formation of proteoglycans found in the structural matrix of joints. Chondroitin sulfates, whether they are absorbed intact or broken into their constituent components, similarly provide additional substrates for the formation of a healthy joint matrix. Evidence also supports the oral administration of chondroitin sulfates for joint disease, both as an agent to slowly reduce symptoms and to reduce the need for non-steroidal anti-inflammatory drugs. The combined use of glucosamine sulfate and chondroitin sulfates in the treatment of degenerative joint disease has become an extremely popular supplementation protocol in arthritic conditions of the joints. Although glucosamine sulfate and chondroitin sulfates are often administered together, there is no information available to demonstrate the combination produces better results than glucosamine sulfate alone.


Distribution of brain sodium accumulation correlates with disability in multiple sclerosis: a cross-sectional 23Na MR imaging study.
To quantify brain sodium accumulations and characterize for the first time the spatial location of sodium abnormalities at different stages of relapsing-remitting (RR) multiple sclerosis (MS) by using sodium 23 ((23)Na) magnetic resonance (MR) imaging. This study was approved by the local committee on ethics, and written informed consent was obtained from all participants. Three-dimensional (23)Na MR imaging data were obtained with a 3.0-T unit in two groups of patients with RR MS-14 with early RR MS (disease duration <5 years) and 12 with advanced RR MS (disease duration >5 years)-and 15 control subjects. Quantitative assessment of total sodium concentration (TSC) levels within compartments (MS lesions, white matter [WM], and gray matter [GM]) as well as statistical mapping analyses of TSC abnormalities were performed. TSC was increased inside demyelinating lesions in both groups of patients, whereas increased TSC was observed in normal-appearing WM and GM only in those with advanced RR MS. In patients, increased TSC inside GM was correlated with disability (as determined with the Expanded Disability Status Scale [EDSS] score; P = .046, corrected) and lesion load at T2-weighted imaging (P = .003, corrected) but not with disease duration (P = .089, corrected). Statistical mapping analysis showed confined TSC increases inside the brainstem, cerebellum, and temporal poles in early RR MS and widespread TSC increases that affected the entire brain in advanced RR MS. EDSS score correlated with TSC increases inside motor networks. TSC accumulation dramatically increases in the advanced stage of RR MS, especially in the normal-appearing brain tissues, concomitant with disability. Brain sodium MR imaging may help monitor the occurrence of tissue injury and disability.


Novel circulating microRNA signature as a potential non-invasive multi-marker test in ER-positive early-stage breast cancer: a case control study.
There are currently no highly sensitive and specific minimally invasive biomarkers for detection of early-stage breast cancer. MicroRNAs (miRNAs) are present in the circulation and may be unique biomarkers for early diagnosis of human cancers. The aim of this study was to investigate the differential expression of miRNAs in the serum of breast cancer patients and healthy controls. Global miRNA analysis was performed on serum from 48 patients with ER-positive early-stage breast cancer obtained at diagnosis (24 lymph node-positive and 24 lymph node-negative) and 24 age-matched healthy controls using LNA-based quantitative real-time PCR (qRT-PCR). A signature of miRNAs was subsequently validated in an independent set of 111 serum samples from 60 patients with early-stage breast cancer and 51 healthy controls and further tested for reproducibility in 3 independent data sets from the GEO Database. A multivariable signature consisting of 9 miRNAs (miR-15a, miR-18a, miR-107, miR-133a, miR-139-5p, miR-143, miR-145, miR-365, miR-425) was identified that provided considerable discrimination between breast cancer patients and healthy controls. Further, the ability of the 9 miRNA signature to stratify samples from breast cancer patients and healthy controls was confirmed in the validation set (p = 0.012) with a corresponding AUC = 0.665 in the ROC-curve analysis. No association between miRNA expression and tumor grade, tumor size, menopausal- or lymph node status was observed. The signature was also successfully validated in a previously published independent data set of circulating miRNAs in early-stage breast cancer (p = 0.024). We present herein a 9 miRNA signature capable of discriminating between ER-positive breast cancer and healthy controls. Using a specific algorithm based on the 9 miRNA signature, the risk for future individuals can be predicted. Since microRNAs are highly stable in blood components, this signature might be useful in the development of a blood-based multi-marker test to improve early detection of breast cancer. Such a test could potentially be used as a screening tool to identify individuals who would benefit from further diagnostic assessment.


The Quest for the Ideal Base: Rational Design of a Nickel Precatalyst Enables Mild, Homogeneous C-N Cross-Coupling.
Palladium-catalyzed amination reactions using soluble organic bases have provided a solution to the many issues associated with heterogeneous reaction conditions. Still, homogeneous C-N cross-coupling approaches cannot yet employ bases as weak and economical as trialkylamines. Furthermore, organic base-mediated methods have not been developed for Ni(0/II) catalysis, despite some advantages of such systems over those employing Pd-based catalysts. We designed a new air-stable and easily prepared Ni(II) precatalyst bearing an electron-deficient bidentate phosphine ligand that enables the cross-coupling of aryl triflates with aryl amines using triethylamine (TEA) as base. The method is tolerant of sterically congested coupling partners, as well as those bearing base- and nucleophile-sensitive functional groups. With the aid of density functional theory (DFT) calculations, we determined that the electron-deficient auxiliary ligands decrease both the pKa of the Ni-bound amine and the barrier to reductive elimination from the resultant Ni(II)-amido complex. Moreover, we determined that the preclusion of Lewis acid-base complexation between the Ni catalyst and the base, due to steric factors, is important for avoiding catalyst inhibition.


Patient and allograft outcomes after kidney transplant for the Indigenous patients in the United States.
The objective is to assess cardiovascular (CV), malignancy, infectious, graft outcomes and tacrolimus levels for the Indigenous patients compared to Whites after kidney transplant (KTx). 165 Indigenous and 165 White patients matched for the KTx year at Mayo Clinic Arizona from 2007-2015 were studied over a median follow-up of 3 years. Propensity score was calculated to account for baseline differences. Compared to Whites, Indigenous patients had the following characteristics: younger age, more obesity, diabetes, hypertension, and required dialysis prior to KTx (p<0.01). Indigenous patients had longer hospital stay for KTx, shorter follow-up and lived further from the transplant center (p<0.05). 210 (63.6%) received deceased donor KTx and more Whites received a living donor KTx compared to Indigenous patients (55.2% vs 17.6%, p<0.0001). Post-KTx, there was no difference in the CV event rates. The cumulative incidence of infectious complications was higher among the Indigenous patients (HR 1.81, p = 0.0005, 48.5% vs 38.2%, p = 0.013), with urinary causes as the most common. Malignancy rates were increased among Whites (13.3% vs 3.0%, p = 0.001) with skin cancer being the most common. There was a significant increase in the dose normalized tacrolimus level for the Indigenous patients compared to Whites at 1 months, 3 months, and 1 year post-KTx. After adjustment for the propensity score, there was no statistical difference in infectious or graft outcomes between the two groups but the mean number of emergency room visits and hospitalizations after KTx was significantly higher for Whites compared to Indigenous patients. Compared to Whites, Indigenous patients have similar CV events, graft outcomes and infectious complications after accounting for baseline differences.


[Treatment of hip fractures in a department of general surgery and a department of orthopedic surgery. A comparison].
All patients treated for fractures of the hip in 1989 in a department of general surgery and a department of orthopedics were included in this study. Data regarding demographics, length of hospitalization, rehabilitation, postoperative complications, types of fractures and modes of operation were retrospectively registered and compared. Mean length of hospitalization was significantly longer and the incidence of postoperative complications was significantly higher in the department of general surgery when compared to the specialized department of orthopedics especially with regard to wound infections and the formation of bedsores. We tentatively propose reasons for these differences.


Soluble CD163-Associated Dietary Patterns and the Risk of Metabolic Syndrome.
Elevated soluble cluster of differentiation 163 (sCD163) concentrations, a marker of macrophage activation, are associated with obesity. Weight reduction decreases circulating CD163 levels, and changes in sCD163 levels are associated with improved metabolic dysfunction. Currently, the relationship between sCD163 and diet remains unclear. This study investigated dietary patterns associated with sCD163 concentrations and its predictive effect on metabolic syndrome (MetS). Data on anthropometrics, blood biochemistry, and a food frequency questionnaire were collected from 166 Taiwanese adults. sCD163 levels independently predicted MetS (odds ratio (OR): 5.35; 95% confidence interval (CI): 2.13~13.44, p < 0.001), non-alcoholic fatty liver disease (OR: 2.19; 95% CI: 1.03~4.64, p < 0.001), and central obesity (OR: 3.90; 95% CI: 1.78~8.55, p < 0.001), after adjusting for age and sex. An adjusted linear regression analysis revealed strong correlations between levels of sCD163 and aspartate transaminase (AST) (β = 0.250 (0.023~0.477), p < 0.05) and red blood cell aggregation (β = 0.332 (0.035~0.628), p < 0.05). sCD163-associated dietary pattern scores (high frequencies of consuming noodles and desserts, and eating at home, and a low intake frequency of steamed/boiled/raw food, white/light-green-colored vegetables, orange/red/purple-colored vegetables, dairy products, seafood, dark-green leafy vegetables, and soy products) were positively correlated with MetS, liver injury biomarkers, and sCD163 levels (all p for trend < 0.05). Individuals with the highest dietary pattern scores (tertile 3) had a 2.37-fold [OR: 2.37; 95% CI: 1.04~5.37, p < 0.05] higher risk of MetS compared to those with the lowest scores (tertile 1). Overall, the study findings suggest the importance of a healthy dietary pattern in preventing elevated sCD163 levels and diet-related chronic disease such as MetS.


Adhesion of Escherichia coli to human uroepithelial cells in vitro.
Optimal conditions for in vitro adherence of Escherichia coli to uroepithelial cells, previously shown to more efficient for strains causing acute symptomatic than that for strains causing "asymptomatic" urinary tract infections, were investigated. Uroepithelial cells from fresh morning urine of healthy individuals and E. coli bacteria from patients with various forms of urinary tract infeciton were used. Adhesion was found to vary, between individuals and epithelial cell types, with epithelial cell viability, bacterial cultivation medium and growth phase, number of bacteria added to the epithelial cells, and incubation time and temperature. Adhesion was also influenced by variations in pH and osmolarity. Optimal test conditions were obtained with post-log-phase bacterial cultures grown on nutrient broth when 10(8) bacteria were added to 10(5) epithelial cells and incubated for 60 min. Considerable variation was found between experiments done on different days, whereas the variation between duplicates was small. The method described may provide a useful tool in the study of the host-parasite relationship in urinary tract infections.


The Role of Gas Exchange Variables in Cardiopulmonary Exercise Testing for Risk Stratification and Management of Heart Failure with Reduced Ejection Fraction.
Heart failure with reduced ejection fraction (HFrEF) is common in the developed world and results in significant morbidity and mortality. Accurate risk assessment methods and prognostic variables are therefore needed to guide clinical decision making for medical therapy and surgical interventions with the ultimate goal of decreasing risk and improving health outcomes. The purpose of this review is to examine the role of cardiopulmonary exercise testing (CPET) and its most commonly used ventilatory gas exchange variables for the purpose of risk stratification and management of HFrEF. We evaluated five widely studied gas exchange variables from CPET in HFrEF patients based on nine previously used systematic criteria for biomarkers. This paper provides clinicians with a comprehensive and critical overview, class recommendations and evidence levels. Although some CPET variables met more criteria than others, evidence supporting the clinical assessment of variables beyond peak V̇O2 is well-established. A multi-variable approach also including the V̇E-V̇CO2 slope and EOV is therefore recommended.


Development and validation of methods for the trace determination of PCBs in biological matrices.
A method was developed to determine trace concentrations of a range of individual PCB congeners in biological samples (grass, silage, cattle faeces and milk-fat) which were taken from rural or 'background' areas of the UK, in order to prepare a mass balance of PCBs in grazing, lactating cows. A simple milk-fat extraction method was compared to Soxhlet extraction and to whole milk extraction. Results indicated that simply boiling milk-fat in hexane with sodium sulphate present gave a comparable extraction of PCBs to other methods. A clean-up method was devised using acid treated silica gel followed by basic alumina chromatography. Grinding frozen grass or silage with sodium sulphate followed by Soxhlet extraction was found to be the most effective method for these matrices, whilst avoiding the potential contamination/loss of PCBs which can be incurred by air, oven or freeze drying. Soxhlet extraction of cattle faeces, after grinding with sodium sulphate, was found to be effective. A rigorous clean-up was devised which involved passing the extracts through silica gel and acid treated silica gel, followed by size exclusion chromatography (gel permeation chromatography, SEC). 13C12 labelled PCBs were used as recovery standards, quantification was performed using GC-MS. A quality control regime and method validation results are presented. The milk analysis method gave within batch mean recoveries of 69-96%, and within batch standard deviations between 1 and 10%. The vegetation analysis method gave within batch mean recoveries of 91-116%, and within batch standard deviations between 1 and 11%. The batch to batch mean recovery for milk analysis was 90%, with an RSD of 2% for high spikes and 5% for low spikes; for vegetation analysis the batch to batch average recovery was 106%, with an RSD of 14% for high spikes and 11% for low spikes. sigma PCB concentrations (53 congeners) of 3900 +/- 790 pg g-1 milk-fat, 1300 +/- 420 pg g-1 dry matter (DM) cattle faeces, 630 +/- 140 pg g-1 DM silage and 1350 +/- 580 pg g-1 DM grass were found during the study.


Comparison of early clinical outcomes following transcatheter aortic valve implantation versus surgical aortic valve replacement versus optimal medical therapy in patients older than 80 years with symptomatic severe aortic stenosis.
Transcatheter aortic valve implantation (TAVI) has become an attractive therapeutic strategy for severe aortic stenosis (AS) in elderly patients due to its minimally-invasive nature. Therefore, early results of its clinical outcomes in elderly Korean patients were evaluated. We compared early clinical outcomes of TAVI, surgical aortic valve replacement (SAVR), and optimal medical therapy (OMT) in patients aged≥80 years with symptomatic severe AS. Treatment groups were allocated as follows: TAVI (n=10), SAVR (n=14), and OMT (n=42). Baseline clinical characteristics including predicted operative mortality were similar among the three groups. However, patients with New York Heart Association functional class III or IV symptoms and smaller aortic valve area were treated with TAVI or SAVR rather than OMT. In-hospital combined safety endpoints (all-cause mortality, major stroke, peri-procedural myocardial infarction, life-threatening bleeding, major vascular complication, and acute kidney injury) after TAVI or SAVR were significantly lower in the TAVI group than in the SAVR group (10.0% vs. 71.4%, respectively, p=0.005), along with an acceptable rate of symptom improvement and device success. During the follow-up period, the TAVI group showed the lowest rate of 3-month major adverse cardiovascular and cerebrovascular events, a composite of all-cause mortality, myocardial infarction, major stroke, and re-hospitalization (TAVI 0.0% vs. SAVR 50.0% vs. OMT 42.9%, p=0.017). Treatment with TAVI was associated with lower event rates compared to SAVR or OMT. Therefore, TAVI may be considered as the first therapeutic strategy in selected patients aged≥80 years with symptomatic severe AS.


Association of body mass index with peripheral arterial disease in older adults: the Cardiovascular Health Study.
The authors hypothesized that the absence of cross-sectional associations of body mass index (BMI; weight (kg)/height (m)(2)) with peripheral arterial disease (PAD) in prior studies may reflect lower weight among persons who smoke or have poor health status. They conducted an observational study among 5,419 noninstitutionalized residents of 4 US communities aged ≥ 65 years at baseline (1989-1990 or 1992-1993). Ankle brachial index was measured, and participants reported their history of PAD procedures. Participants were followed longitudinally for adjudicated incident PAD events. At baseline, mean BMI was 26.6 (standard deviation, 4.6), and 776 participants (14%) had prevalent PAD. During 13.2 (median) years of follow-up through June 30, 2007, 276 incident PAD events occurred. In cross-sectional analysis, each 5-unit increase in BMI was inversely associated with PAD (prevalence ratio (PR) = 0.92, 95% confidence interval (CI): 0.85, 1.00). However, among persons in good health who had never smoked, the direction of association was opposite (PR = 1.20, 95% CI: 0.94, 1.52). Similar results were observed between BMI calculated using weight at age 50 years and PAD prevalence (PR = 1.30, 95% CI: 1.11, 1.51) and between BMI at baseline and incident PAD events occurring during follow-up (hazard ratio = 1.32, 95% CI: 1.00, 1.76) among never smokers in good health. Greater BMI is associated with PAD in older persons who remain healthy and have never smoked. Normal weight maintenance may decrease PAD incidence and associated comorbidity in older age.


Tissue factor and cancer stem cells: is there a linkage?
A common feature in the progression of multiple human malignancies is the protracted deregulation of the coagulation system, often referred to as cancer coagulopathy. Indeed, cancer cells and their vascular stroma often exhibit procoagulant properties, of which deregulation of tissue factor (TF) expression is a notable, although not the sole example. These changes can be traced to oncogenic influences affecting epidermal growth factor receptor (EGFR), EGFRvIII, K-ras, p53, PTEN, and probably many other proto-oncogenes and tumor suppressors in tumor parenchyma. Cancer stem cells (CSCs)/tumor initiating cells (TICs) are thought to represent the primary target and the main cellular effector through which oncogenic mutations exert their tumor-inducing effects. In so doing, CSCs/TICs depend on interactions with the tumor vasculature, which forms supportive niches for their clonal growth. We postulate that TF contributes to these interactions (directly or indirectly) through procoagulant and signaling effects, the latter executed in concert with juxtaposed protease activated receptors (mainly PAR-1 and PAR-2). TF/PAR system acts as a "blood sensing" mechanism, whereby cancer cells, including CSCs/TICs, may respond to plasma proteases (Factors VIIa, Xa, and IIa) and their related microenvironmental changes (fibrin deposition, activation of platelets). A growing body of still largely circumstantial evidence suggests that these events may contribute to the CSC/TIC niche, which could influence tumor initiation, metastasis, recurrence, and therapeutic intractability. Indeed, certain types of cancer cells harboring markers of CSCs (CD133) exhibit elevated TF expression and depend on this receptor to efficiently initiate tumor growth. We propose that both tumor cell-associated and host-related TF could influence the properties of CSCs, and that agents targeting the TF/PAR system may represent a hitherto unappreciated therapeutic opportunity to control cancer progression by influencing the CSC/TIC compartment.


Diffusion tensor imaging in patients with essential tremor.
The traditional paradigm has regarded essential tremor (ET) as a benign disorder. However, recent clinical, neuroimaging, and neuropathologic studies suggest that ET may be a progressive neurologic disorder. Based on clinicopathologic findings that cerebellum and its outflow are the key structures in ET and degeneration of gray matter in cerebellum is followed by consequent wallerian degeneration of white matter (WM) fibers, the aim of the present study was to investigate changes in anisotropy in patients with ET. Fractional anisotropy (FA) images were generated from DTI data acquired at 1.5T in 10 patients with ET compared with 8 control subjects by using statistical parametric mapping to make voxel-by-voxel comparisons. Compared with the control subjects, the patients with ET exhibited significantly reduced FA (P(uncorrected) < .005) in the anterolateral portion of the right pons and decreased FA in the bilateral cerebellum, left retrorubral area of the midbrain, and bilateral deep WM, including the orbitofrontal, lateral frontal, parietal, and temporal WM. This study demonstrates that structural changes in the WM are extensive in patients with ET, supporting the findings of previous functional neuroimaging and pathologic studies.


LP99: Discovery and Synthesis of the First Selective BRD7/9 Bromodomain Inhibitor.
The bromodomain-containing proteins BRD9 and BRD7 are part of the human SWI/SNF chromatin-remodeling complexes BAF and PBAF. To date, no selective inhibitor for BRD7/9 has been reported despite its potential value as a biological tool or as a lead for future therapeutics. The quinolone-fused lactam LP99 is now reported as the first potent and selective inhibitor of the BRD7 and BRD9 bromodomains. Development of LP99 from a fragment hit was expedited through balancing structure-based inhibitor design and biophysical characterization against tractable chemical synthesis: Complexity-building nitro-Mannich/lactamization cascade processes allowed for early structure-activity relationship studies whereas an enantioselective organocatalytic nitro-Mannich reaction enabled the synthesis of the lead scaffold in enantioenriched form and on scale. This epigenetic probe was shown to inhibit the association of BRD7 and BRD9 to acetylated histones in vitro and in cells. Moreover, LP99 was used to demonstrate that BRD7/9 plays a role in regulating pro-inflammatory cytokine secretion.


[A network RER rooted on in vitro readout assays of Plasmodium falciparum sensitivity to chloroquine in the Indian Ocean Region].
Chloroquine has been used as a first line drug to treat uncomplicated malaria cases during the last five decades in Madagascar and in the Comoros Union. The four plasmodial species known to infect humans occur on Madagascar Island. Chloroquine-resistant malaria cases, sometimes only suspected from presumptive malaria cases, have been reported in both countries. Thus, to redefine a strategy and a policy to cure malaria, there is a need to get relevant and updated data. In December 1999, the Madagascan Ministry of Health and the Institut Pasteur de Madagascar formed a network named RER for malaria resistance surveillance. In 2000 and 2001, 18 study sites (17 throughout Madagascar and 1 in Comoros) joined this network. Health-care workers were trained mainly for malaria diagnosis through the use of blood smear examination and for malaria case management. To alleviate the lack of competent medical teams within the health centres, and for technical and logistic reasons, as part of the network activities, it was decided to start with in vitro tests to assess the sensitivity of P.falciparum isolates to chloroquine by means of the isotopic method. Parasitized blood samples were collected from consenting patients. P.falciparum isolates were more predominant (989/1,036). Out of the 564 tests done, 432 (76.6%) could be assessed. Results demonstrated that 94.3% (381/404) of the Madagascan P.falciparum isolates were susceptible to chloroquine. In contrast, chloroquine-resistant isolates were prevalent in Comoros (8/28). The network set-up is presented. The usefulness of the in vivo approach and of the in vitro investigations (chemosusceptibility test and screening of mutations accounting for resistance to chloroquine) to monitor the emergence and the dissemination of chloroquine-resistant parasites is discussed.


Amantadine and rimantadine for preventing and treating influenza A in adults.
Amantadine hydrochloride and rimantadine hydrochloride have antiviral properties, but they are not widely used due to a lack of knowledge of their potential value and concerns about possible adverse effects. The objective of this review was to assess the effects and safety of amantadine and rimantadine in healthy adults. We searched the Cochrane Controlled Trials Register, MEDLINE, EMBASE and reference lists of articles. We also contacted manufacturers, researchers and authors. Randomised and quasi-randomised studies comparing amantadine and/or rimantadine with placebo, control antivirals or no intervention, or comparing doses or schedules of amantadine and/or rimantadine in healthy adults. For prevention trials the numbers of participants with clinically defined influenza, with serologically confirmed clinical influenza A and adverse effects were analysed. Analysis for treatment trials was of the mean duration of fever and adverse effects. Amantadine prevented 23% of clinical influenza cases (95% confidence interval 11% to 34%), and 63% of serologically confirmed clinical influenza A cases (95% confidence interval 42% to 76%). Amantadine reduced duration of fever by one day (95% confidence interval 0.7 to 1.3). Rimantadine demonstrated comparable effectiveness, but there were fewer trials and the results for prevention were not statistically significant. Both amantadine and rimantadine induced significant gastrointestinal adverse effects. Adverse effects of the central nervous system and study withdrawals were significantly more common with amantadine than rimantadine. Amantadine and rimantadine have comparable effectiveness in the prevention and treatment of influenza A in healthy adults, although rimantadine induces fewer adverse effects than amantadine. [This abstract has been prepared centrally.]


Neighborhood adversity, child health, and the role for community development.
Despite medical advances, childhood health and well-being have not been broadly achieved due to rising chronic diseases and conditions related to child poverty. Family and neighborhood living conditions can have lasting consequences for health, with community adversity affecting health outcomes in significant part through stress response and increased allostatic load. Exposure to this "toxic stress" influences gene expression and brain development with direct and indirect negative consequences for health. Ensuring healthy child development requires improving conditions in distressed, high-poverty neighborhoods by reducing children's exposure to neighborhood stressors and supporting good family and caregiver functioning. The community development industry invests more than $200 billion annually in low-income neighborhoods, with the goal of improving living conditions for residents. The most impactful investments have transformed neighborhoods by integrating across sectors to address both the built environment and the social and service environment. By addressing many facets of the social determinants of health at once, these efforts suggest substantial results for children, but health outcomes generally have not been considered or evaluated. Increased partnership between the health sector and community development can bring health outcomes explicitly into focus for community development investments, help optimize intervention strategies for health, and provide natural experiments to build the evidence base for holistic interventions for disadvantaged children. The problems and potential solutions are beyond the scope of practicing pediatricians, but the community development sector stands ready to engage in shared efforts to improve the health and development of our most at-risk children.


Consistent cooperation in a cichlid fish is caused by maternal and developmental effects rather than heritable genetic variation.
Studies on the evolution of cooperative behaviour are typically confined to understanding its adaptive value. It is equally essential, however, to understand its potential to evolve, requiring knowledge about the phenotypic consistency and genetic basis of cooperative behaviour. While previous observational studies reported considerably high heritabilities of helping behaviour in cooperatively breeding vertebrates, experimental studies disentangling the relevant genetic and non-genetic components of cooperative behaviour are lacking. In a half-sibling breeding experiment, we investigated the repeatability and heritability of three major helping behaviours performed by subordinates of the cooperatively breeding fish Neolamprologus pulcher To experimentally manipulate the amount of help needed in a territory, we raised the fish in two environments differing in egg predation risk. All three helping behaviours were significantly repeatable, but had very low heritabilities. The high within-individual consistencies were predominantly due to maternal and permanent environment effects. The perceived egg predation risk had no effect on helping, but social interactions significantly influenced helping propensities. Our results reveal that developmentally plastic adjustments of provided help to social context shape cooperative phenotypes, whereas heritable genetic variation plays a minor role.


Fluoride substitution of vitamin D analogs at C-26 and C-27: enhancement of activity of 25-hydroxyvitamin D but not of 1,25-dihydroxyvitamin D on bone and intestine in vitro.
A series of analogs of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] with multiple fluorine substitutions in positions 26 and 27 have been tested for their activities 1) in competing with 1,25-(OH)2D3 for binding to partially purified chick intestinal cytosol, 2) in stimulating resorption of fetal rat limb bones in vitro and 3) in competing with 25-hydroxyvitamin D3 for binding sites in rat plasma. The relative potencies of 26,26,26,27,27,27-hexafluoro-25-hydroxyvitamin D3, 26,26,26-trifluoro-25-hydroxyvitamin D3, 27-nor-26,26,26-trifluoro-25-hydroxyvitamin D3 and 25-hydroxyvitamin D3 in competing for intestinal cytosolic binding were 17:11:1:1. The relative potencies of the same series of compounds on stimulating resorption of fetal rat bones was 25:21:9:1. The relative ability of these four compounds to compete for plasma binding sites was 0.3:0.3:0.4:1.0. The results indicate that multiple fluorine substitution in the vicinity of the 25-hydroxyl group can markedly enhance the direct effects of 25-hydroxyvitamin D3 on its target tissues. This is postulated to result from the electronegativity of the fluorines which increases the acidity of the 25-hydroxyl group and enhances its affinity for the receptor. In contrast to the effects seen with the 25-hydroxyvitamin D3 analogs, multiple fluorine substitution in positions 26 and 27 did not enhance the activity of 1,25-(OH)2D3 on either cytosolic binding or bone resorption. Presumably, this is because biological activity, expressed in terms of affinity for the receptor, is already optimal in the 1,25-(OH)2D3 structure. The relative activities of the 26,27-hexafluoro derivative and 1,25-(OH)2D3 in competition for the plasma binding sites was 0.4:1.0.(ABSTRACT TRUNCATED AT 250 WORDS)


The Arabidopsis condensin CAP-D subunits arrange interphase chromatin.
Condensins are best known for their role in shaping chromosomes. Other functions such as organizing interphase chromatin and transcriptional control have been reported in yeasts and animals, but little is known about their function in plants. To elucidate the specific composition of condensin complexes and the expression of CAP-D2 (condensin I) and CAP-D3 (condensin II), we performed biochemical analyses in Arabidopsis. The role of CAP-D3 in interphase chromatin organization and function was evaluated using cytogenetic and transcriptome analysis in cap-d3 T-DNA insertion mutants. CAP-D2 and CAP-D3 are highly expressed in mitotically active tissues. In silico and pull-down experiments indicate that both CAP-D proteins interact with the other condensin I and II subunits. In cap-d3 mutants, an association of heterochromatic sequences occurs, but the nuclear size and the general histone and DNA methylation patterns remain unchanged. Also, CAP-D3 influences the expression of genes affecting the response to water, chemicals, and stress. The expression and composition of the condensin complexes in Arabidopsis are similar to those in other higher eukaryotes. We propose a model for the CAP-D3 function during interphase in which CAP-D3 localizes in euchromatin loops to stiffen them and consequently separates centromeric regions and 45S rDNA repeats.


What happens to medical patients with psychiatric disorder?
Medical, psychiatric and social outcome were examined in medical in-patients previously identified as suffering from psychiatric disorder. One third of patients with an affective (emotional) disorder on admission were still psychiatrically ill four months after discharge. Persistent disorder was associated with continuing physical illness. During the year following admission those with affective disorder on admission continued to make greater demands on medical, social and psychiatric services than matched controls and had double the mortality rate (not significant). Patients with organic mental states on admission had a high mortality and morbidity, and made considerable continuing use of general hospital social and psychiatric services. Improved recognition of psychiatric disorder during hospital admission could result in better overall care of medical patients' psychiatric and social difficulties and more effective use of medical resources.


Drugs, the brain, and behavior: a graduate student-run comprehensive course in neuroscience.
Drugs, the Brain, and Behavior is an interdisciplinary two-semester upper level course at Georgetown University designed to expose undergraduate and graduate students to broad areas of the neurosciences, to promote the development of scientific literacy in these students, and to provide pedagogical experience for Ph.D. students in the Interdisciplinary Program in Neuroscience (IPN) at all stages of training. Drugs, the Brain, and Behavior fulfills these goals through a unique model of student-teaching. This lecture-based, team-taught course is completely run and taught by Ph.D. students in the IPN. It is designed to gradually increase the teaching duties of new instructors, providing a structured setting for them to develop their pedagogical skills. We encourage scientific literacy in our students through the incorporation of primary literature and experimental results throughout the course. The strategies we have employed have increased student confidence on a variety of measures of scientific literacy. While running a team-taught course, we have also developed several strategies for coordinating team-taught courses within semesters and across years, which could easily be adapted to other courses.


Competition between pentoses and glucose during uptake and catabolism in recombinant Saccharomyces cerevisiae.
In mixed sugar fermentations with recombinant Saccharomyces cerevisiae strains able to ferment D-xylose and L-arabinose the pentose sugars are normally only utilized after depletion of D-glucose. This has been attributed to competitive inhibition of pentose uptake by D-glucose as pentose sugars are taken up into yeast cells by individual members of the yeast hexose transporter family. We wanted to investigate whether D-glucose inhibits pentose utilization only by blocking its uptake or also by interfering with its further metabolism. To distinguish between inhibitory effects of D-glucose on pentose uptake and pentose catabolism, maltose was used as an alternative carbon source in maltose-pentose co-consumption experiments. Maltose is taken up by a specific maltose transport system and hydrolyzed only intracellularly into two D-glucose molecules. Pentose consumption decreased by about 20 - 30% during the simultaneous utilization of maltose indicating that hexose catabolism can impede pentose utilization. To test whether intracellular D-glucose might impair pentose utilization, hexo-/glucokinase deletion mutants were constructed. Those mutants are known to accumulate intracellular D-glucose when incubated with maltose. However, pentose utilization was not effected in the presence of maltose. Addition of increasing concentrations of D-glucose to the hexo-/glucokinase mutants finally completely blocked D-xylose as well as L-arabinose consumption, indicating a pronounced inhibitory effect of D-glucose on pentose uptake. Nevertheless, constitutive overexpression of pentose-transporting hexose transporters like Hxt7 and Gal2 could improve pentose consumption in the presence of D-glucose. Our results confirm that D-glucose impairs the simultaneous utilization of pentoses mainly due to inhibition of pentose uptake. Whereas intracellular D-glucose does not seem to have an inhibitory effect on pentose utilization, further catabolism of D-glucose can also impede pentose utilization. Nevertheless, the results suggest that co-fermentation of pentoses in the presence of D-glucose can significantly be improved by the overexpression of pentose transporters, especially if they are not inhibited by D-glucose.


Role of the protein tyrosine phosphatase Shp2 in homeostasis of the intestinal epithelium.
Protein tyrosine phosphorylation is thought to be important for regulation of the proliferation, differentiation, and rapid turnover of intestinal epithelial cells (IECs). The role of protein tyrosine phosphatases in such homeostatic regulation of IECs has remained largely unknown, however. Src homology 2-containing protein tyrosine phosphatase (Shp2) is a ubiquitously expressed cytoplasmic protein tyrosine phosphatase that functions as a positive regulator of the Ras-mitogen-activated protein kinase (MAPK) signaling pathway operative downstream of the receptors for various growth factors and cytokines, and it is thereby thought to contribute to the regulation of cell proliferation and differentiation. We now show that mice lacking Shp2 specifically in IECs (Shp2 CKO mice) develop severe colitis and die as early as 3 to 4 weeks after birth. The number of goblet cells in both the small intestine and colon of Shp2 CKO mice was markedly reduced compared with that for control mice. Furthermore, Shp2 CKO mice showed marked impairment of both IEC migration along the crypt-villus axis in the small intestine and the development of intestinal organoids from isolated crypts. The colitis as well as the reduction in the number of goblet cells apparent in Shp2 CKO mice were normalized by expression of an activated form of K-Ras in IECs. Our results thus suggest that Shp2 regulates IEC homeostasis through activation of Ras and thereby protects against the development of colitis.


Innovative and diverse strategies toward primary health care reform: lessons learned from the Canadian experience.
In the last decade, Canadian provincial and territorial health systems have taken diverse approaches to strengthening primary care delivery. Although the Canadian and US systems differ in significant ways, important commonalities include the organization of care delivery, core principles guiding primary care reform, and some degree of provincial/state autonomy. This suggests that Canadian experiences, which employed a variety of tools, strategies, and policies, may be informative for US efforts to improve primary care. The range of primary care reform initiatives implemented across Canada target organizational infrastructure, provider payment, health care workforce, and quality and safety. Primary care teams and networks in which multiple physicians work in concert with other providers have become widespread in some provinces; they vary on a number of dimensions, including physician payment, incorporation of other providers, and formal enrolment of patients. Family medicine is attracting more recent medical school graduates, a trend likely affected by new physician payment models, increases in the number of primary care providers, and efforts to better integrate nonphysician providers into clinical practice. Efforts to integrate electronic medical records into practice and pursue quality improvement strategies are gaining ground in some provinces. Canadian primary care reform initiatives rely on voluntary participation, incremental change, and diverse models, encouraging engagement and collaboration from a range of stakeholders including patients, providers, and policymakers. Cross-country collaboration in evaluating and translating Canada's primary care reform efforts are likely to yield important lessons for the US experience.


Low-dose infliximab (3 mg/kg) significantly reduces spinal inflammation on magnetic resonance imaging in patients with ankylosing spondylitis: a randomized placebo-controlled study.
To evaluate the influence of low-dose infliximab (IFX) on spinal inflammation scored by magnetic resonance imaging (MRI). The dose recommended for rheumatoid arthritis (3 mg/kg) is also clinically effective for ankylosing spondylitis (AS), although effects on spinal inflammation as defined by MRI have yet to be described in a placebo-controlled trial. In a 12-week double-blind period, patients were randomized 1:1 to receive either IFX 3 mg/kg at 0, 2, and 6 weeks, or placebo. Spinal inflammation in discovertebral units (DVU) was measured by the Spondyloarthritis Research Consortium of Canada (SPARCC) MRI Index at baseline and 12 weeks by 3 readers blinded to timepoint and treatment allocation. We also compared reliability and discrimination of the SPARCC MRI index based on evaluation of the entire spine (23 DVU score) compared to assessment of only the 6 most severely affected DVU (6 DVU score). At Week 12, patients treated with IFX experienced mean reductions of 55.1% and 57.2% in the 6 DVU and 23 DVU SPARCC scores, respectively, compared with a mean increase of 5.8% and decrease of 3.4% in 6 DVU and 23 DVU scores, respectively, for patients taking placebo (p < 0.001). A large treatment effect (Guyatt's effect size >or= 1.7) and high reliability was evident and comparable between 6 DVU and 23 DVU scoring methods. Treatment with low-dose IFX leads to a large treatment effect on spinal inflammation as measured by MRI. Scoring for inflammation of only the most severely affected regions of the spine by MRI is comparable to assessment of the entire spine.


What kinds of things are psychiatric disorders?
This essay explores four answers to the question 'What kinds of things are psychiatric disorders?' Essentialist kinds are classes whose members share an essence from which their defining features arise. Although elegant and appropriate for some physical (e.g. atomic elements) and medical (e.g. Mendelian disorders) phenomena, this model is inappropriate for psychiatric disorders, which are multi-factorial and 'fuzzy'. Socially constructed kinds are classes whose members are defined by the cultural context in which they arise. This model excludes the importance of shared physiological mechanisms by which the same disorder could be identified across different cultures. Advocates of practical kinds put off metaphysical questions about 'reality' and focus on defining classes that are useful. Practical kinds models for psychiatric disorders, implicit in the DSM nosologies, do not require that diagnoses be grounded in shared causal processes. If psychiatry seeks to tie disorders to etiology and underlying mechanisms, a model first proposed for biological species, mechanistic property cluster (MPC) kinds, can provide a useful framework. MPC kinds are defined not in terms of essences but in terms of complex, mutually reinforcing networks of causal mechanisms. We argue that psychiatric disorders are objectively grounded features of the causal structure of the mind/brain. MPC kinds are fuzzy sets defined by mechanisms at multiple levels that act and interact to produce the key features of the kind. Like species, psychiatric disorders are populations with central paradigmatic and more marginal members. The MPC view is the best current answer to 'What kinds of things are psychiatric disorders?'


Ambulatory function in spinal muscular atrophy: Age-related patterns of progression.
Individuals with spinal muscular atrophy (SMA) type 3 are able to walk but they have weakness, gait impairments and fatigue. Our primary study objective was to examine longitudinal changes in the six-minute walk test (6MWT) and to evaluate whether age and SMA type 3 subtype are associated with decline in ambulatory function. Data from three prospective natural history studies were used. Seventy-three participants who performed the 6MWT more than once, at least 6 months apart, were included; follow-up ranged from 0.5-9 years. Only data from patients who completed the 6MWT were included. The mean age of the participants was 13.5 years (range 2.6-49.1), with 52 having disease onset before age 3 years (type 3A). At baseline, type 3A participants walked a shorter distance on average (257.1 m) than type 3B participants (390.2 m) (difference = 133.1 m, 95% confidence interval [CI] 71.8-194.3, p < 0.001). Distance walked was weakly associated with age (r = 0.25, p = 0.04). Linear mixed effects models were used to estimate the mean annual rate of change. The overall mean rate of change was -7.8 m/year (95% CI -13.6 --2.0, p = 0.009) and this did not differ by subtype (type 3A: -8.5 m/year, type 3B: -6.6 m/year, p = 0.78), but it did differ by age group (< 6: 9.8 m/year; 6-10: -7.9 m/year; 11-19: -20.8 m/year; ≥ 20: -9.7 m/year; p = 0.005). Our results showed an overall decline on the 6MWT over time, but different trajectories were observed depending on age. Young ambulant SMA patients gain function but in adolescence, patients lose function. Future clinical trials in ambulant SMA patients should consider in their design the different trajectories of ambulatory function over time, based on age.


Substitution of the premembrane and envelope protein genes of Modoc virus with the homologous sequences of West Nile virus generates a chimeric virus that replicates in vertebrate but not mosquito cells.
Most known flaviviruses, including West Nile virus (WNV), are maintained in natural transmission cycles between hematophagous arthropods and vertebrate hosts. Other flaviviruses such as Modoc virus (MODV) and Culex flavivirus (CxFV) have host ranges restricted to vertebrates and insects, respectively. The genetic elements that modulate the differential host ranges and transmission cycles of these viruses have not been identified. Fusion polymerase chain reaction (PCR) was used to replace the capsid (C), premembrane (prM) and envelope (E) genes and the prM-E genes of a full-length MODV infectious cDNA clone with the corresponding regions of WNV and CxFV. Fusion products were directly transfected into baby hamster kidney-derived cells that stably express T7 RNA polymerase. At 4 days post-transfection, aliquots of each supernatant were inoculated onto vertebrate (BHK-21 and Vero) and mosquito (C6/36) cells which were then assayed for evidence of viral infection by reverse transcription-PCR, Western blot and plaque assay. Chimeric virus was recovered in cells transfected with the fusion product containing the prM-E genes of WNV. The virus could infect vertebrate but not mosquito cells. The in vitro replication kinetics and yields of the chimeric virus were similar to MODV but the chimeric virus produced larger plaques. Chimeric virus was not recovered in cells transfected with any of the other fusion products. Our data indicate that genetic elements outside of the prM-E gene region of MODV condition its vertebrate-specific phenotype.


Experimental demonstration of intermodal nonlinear effects between full vectorial modes in a few moded fiber.
We experimentally investigate intermodal nonlinear interactions, such as Raman scattering and four wave mixing. The fiber used is a specially designed few moded fiber, which splits the degeneracy of the first mode group, leading to stable propagation of the two full vectorial modes, TM01 and TE01. For the Raman experiments pumping occur in either the fundamental mode or the two full vectorial modes, whereas the signal is in the fundamental mode. In all three experiments approximately 40 dB of gain is achieved using 307 W of pump peak power. When pumping in either of the full vectorial modes four wave mixing is observed.


Pro-anorexia and pro-recovery photo sharing: a tale of two warring tribes.
There is widespread use of the Internet to promote anorexia as a lifestyle choice. Pro-anorexia content can be harmful for people affected or at risk of having anorexia. That movement is actively engaged in sharing photos on social networks such as Flickr. To study the characteristics of the online communities engaged in disseminating content that encourages eating disorders (known as "pro-anorexia") and to investigate if the posting of such content is discouraged by the posting of recovery-oriented content. The extraction of pro-anorexia and pro-recovery photographs from the photo sharing site Flickr pertaining to 242,710 photos from 491 users and analyzing four separate social networks therein. Pro-anorexia and pro-recovery communities interact to a much higher degree among themselves than what is expected from the distribution of contacts (only 59-72% of contacts but 74-83% of comments are made to members inside the community). Pro-recovery users employ similar words to those used by pro-anorexia users to describe their photographs, possibly in order to ensure that their content appears when pro-anorexia users search for images. Pro-anorexia users who are exposed to comments from the opposite camp are less likely to cease posting pro-anorexia photographs than those who do not receive such comments (46% versus 61%), and if they cease, they do so approximately three months later. Our observations show two highly active communities, where most interaction is within each community. However, the pro-recovery community takes steps to ensure that their content is visible to the pro-anorexia community, both by using textual descriptions of their photographs that are similar to those used by the pro-anorexia group and by commenting to pro-anorexia content. The latter activity is, however, counterproductive, as it entrenches pro-anorexia users in their stance. Our results highlight the nature of pro-anorexia and pro-recovery photo sharing and accentuate the need for clinicians to be aware of such content and its effect on their patients. Our findings suggest that some currently used interventions are not useful in helping pro-anorexia users recover. Thus, future work should focus on new intervention methods, possibly tailored to individual characteristics.


Advances in the development of universal influenza vaccines.
Despite the widespread availability and use of influenza vaccines, influenza still poses a considerable threat to public health. Vaccines against seasonal influenza do not offer protection against pandemic viruses, and vaccine efficacy against seasonal viruses is reduced in seasons when the vaccine composition is not a good match for the predominant circulating viruses. Vaccine efficacy is also reduced in older adults, who are one of the main target groups for vaccination. The continual threat of pandemic influenza, with the known potential for rapid spread around the world and high mortality rates, has prompted researchers to develop a number of novel approaches to providing immunity to this virus, focusing on target antigens which are highly conserved between different influenza A virus subtypes. Several of these have now been taken into clinical development, and this review discusses the progress that has been made, as well as considering the requirements for licensing these new vaccines and how they might be used in the future.


Stenting of the right ventricular outflow tract.
To assess the indication, technical aspects, and outcome of stenting of the right ventricular outflow tract (RVOT) in the management of symptomatic patients with severely limited pulmonary blood flow. Retrospective case note and procedure review of patients undergoing stenting of the RVOT over an 8 year period. Between 2005 and 2012, 52 selected patients underwent percutaneous stent implantation into a very narrow RVOT to improve pulmonary blood flow. Median age at stent implantation was 63 (range 4-406) days and median weight was 3.8 (1.7-12.2) kg. 52 patients underwent stent implantation. Median procedure time was 57 (24-260) min and fluoroscopy time 16 (5.5-73) min. There was one procedural death (1.9%) and one emergency surgery (1.9%). Saturations increased from 71% (52-83%) to 92% (81-100%) (p<0.001). Two patients required early shunts due to inadequate palliation and two died from non-cardiac causes. Sixteen further catheter interventions were undertaken (balloon in 7, further stent in 9). Twenty-nine patients underwent delayed surgery (complete repair in 26, palliative in 3) at a median of 172 (52-758) days post-stenting. Left pulmonary artery Z score increased from a pre-interventional value of -1.75 (-4.96 to 0.67) to a pre-surgical value of -0.55 (-4.12 to 1.97), (p<0.01). Median right pulmonary artery Z score increased from -2.63 (-7.70 to 0.89) to -0.75 (-6.69 to 1.18) (p<0.01) . Seventeen patients remain well palliated after a median of 122 (40-286) days. Stenting of the RVOT is an effective treatment option in the initial management of selected patients with very reduced pulmonary blood flow.


The heat released during catalytic turnover enhances the diffusion of an enzyme.
Recent studies have shown that the diffusivity of enzymes increases in a substrate-dependent manner during catalysis. Although this observation has been reported and characterized for several different systems, the precise origin of this phenomenon is unknown. Calorimetric methods are often used to determine enthalpies from enzyme-catalysed reactions and can therefore provide important insight into their reaction mechanisms. The ensemble averages involved in traditional bulk calorimetry cannot probe the transient effects that the energy exchanged in a reaction may have on the catalyst. Here we obtain single-molecule fluorescence correlation spectroscopy data and analyse them within the framework of a stochastic theory to demonstrate a mechanistic link between the enhanced diffusion of a single enzyme molecule and the heat released in the reaction. We propose that the heat released during catalysis generates an asymmetric pressure wave that results in a differential stress at the protein-solvent interface that transiently displaces the centre-of-mass of the enzyme (chemoacoustic effect). This novel perspective on how enzymes respond to the energy released during catalysis suggests a possible effect of the heat of reaction on the structural integrity and internal degrees of freedom of the enzyme.


Referral to yoga therapists in rural primary health care: A survey of general practitioners in rural and regional New South Wales, Australia.
Yoga is an increasingly accepted complementary treatment modality for referral in Australian general practice, yet this practitioner group has largely escaped research attention in Australia. Complementary medicine use is highest in rural and regional areas, where a number of primary health care challenges are also more pronounced. Despite the significant role of complementary therapists in rural and regional Australia, and the increasing acceptance of yoga therapy in general practice, there has been little exploration of the interface between yoga therapists and conventional primary health care practitioners in this area. A 27-item questionnaire was sent to all 1486 general practitioners (GPs) currently practising in rural and regional Divisions of General Practice in New South Wales, Australia. Completed questionnaires were returned by 585 GPs, with 49 returned as 'no longer at this address' (response rate 40.7%). One-in-eight GPs (12.1%) advised their patients of specific yoga therapies and protocols, and 7.2% advised specific meditation techniques. Three-quarters of GPs (76.6%) referred to a yoga therapist at least a few times per year, with 12.5% of GPs referring at least once per week. GPs being in a remote location (OR = 10.95; CI: 1.55, 77.31), being female (OR = 1.85; 95% CI: 1.16, 2.94), GPs graduating from an Australian medical school (OR = 4.52; 95% CI: 2.61, 7.80), perceiving lack of other treatment options (OR = 3.29; 95% CI: 1.61, 6.74), GPs reporting good or very good knowledge of yoga therapies (OR = 18.2; 95% CI: 9.19, 36.19), and GPs using CAM for their own personal health (OR = 4.53; 95% CI: 2.60, 7.87) were all independently predictive of increased referral to yoga therapists amongst the rural GPs in this study. There is a significant interface between yoga therapists in Australian rural and regional general practice. There is generally high support for yoga therapies among Australian GPs, with low levels of opposition to the incorporation of these therapies in patient care. There is a need for increased research into yoga therapies practice, policy and regulation in these areas.


Perspective: Quantum Hamiltonians for optical interactions.
The multipolar Hamiltonian of quantum electrodynamics is extensively employed in chemical and optical physics to treat rigorously the interaction of electromagnetic fields with matter. It is also widely used to evaluate intermolecular interactions. The multipolar version of the Hamiltonian is commonly obtained by carrying out a unitary transformation of the Coulomb gauge Hamiltonian that goes by the name of Power-Zienau-Woolley (PZW). Not only does the formulation provide excellent agreement with experiment, and versatility in its predictive ability, but also superior physical insight. Recently, the foundations and validity of the PZW Hamiltonian have been questioned, raising a concern over issues of gauge transformation and invariance, and whether observable quantities obtained from unitarily equivalent Hamiltonians are identical. Here, an in-depth analysis of theoretical foundations clarifies the issues and enables misconceptions to be identified. Claims of non-physicality are refuted: the PZW transformation and ensuing Hamiltonian are shown to rest on solid physical principles and secure theoretical ground.


Plasma vitamin C predicts incident heart failure in men and women in European Prospective Investigation into Cancer and Nutrition-Norfolk prospective study.
Fruit and vegetable intake has been associated with lower risk for cardiovascular risk factors and disease, but data on heart failure are sparse and inconsistent. The association of plasma vitamin C, a biomarker reflecting fruit and vegetable intake, with heart failure has not been studied. We examined the prospective association of plasma vitamin C concentrations with incident fatal and nonfatal heart failure events in apparently healthy 9,187 men and 11,112 women aged 39 to 79 years participating in the "European Prospective Investigation into Cancer and Nutrition" study in Norfolk. The risk of heart failure decreased with increasing plasma vitamin C; the hazard ratios comparing each quartile with the lowest were 0.76 (95% CI 0.65-0.88), 0.70 (95% CI 0.60-0.81), and 0.62 (95% CI 0.53-0.74) in age- and sex-adjusted analyses (P for trend <.0001). Every 20 μmol/L increase in plasma vitamin C concentration (1 SD) was associated with a 9% relative reduction in risk of heart failure after adjustment for age, sex, smoking, alcohol consumption, physical activity, occupational social class, educational level, systolic blood pressure, diabetes, cholesterol concentration, and body mass index, with similar result if adjusting for interim coronary heart disease. Plasma vitamin C, a biomarker reflecting fruit and vegetable intake, was inversely associated with the risk of heart failure in this healthy population. This observation should be regarded as hypothesis generating for further prospective trials aimed at examining the effect of a diet rich in fruit and vegetables for prevention of heart failure.


Recombinant viral vaccines expressing merozoite surface protein-1 induce antibody- and T cell-mediated multistage protection against malaria.
Protecting against both liver and blood stages of infection is a long-sought goal of malaria vaccine design. Recently, we described the use of replication-defective viral vaccine vectors expressing the malaria antigen merozoite surface protein-1 (MSP-1) as an antimalarial vaccine strategy that elicits potent and protective antibody responses against blood-stage parasites. Here, we show that vaccine-induced MSP-1-specific CD4(+) T cells provide essential help for protective B cell responses, and CD8(+) T cells mediate significant antiparasitic activity against liver-stage parasites. Enhanced survival is subsequently seen in immunized mice following challenge with sporozoites, which mimics the natural route of infection more closely than when using infected red blood cells. This effect is evident both in the presence and absence of protective antibodies and is associated with decreased parasite burden in the liver followed by enhanced induction of the cytokine IFN-gamma in the serum. Multistage immunity against malaria can thus be achieved by using viral vectors recombinant for MSP-1.


A fast algorithm for robust mixtures in the presence of measurement errors.
In experimental and observational sciences, detecting atypical, peculiar data from large sets of measurements has the potential of highlighting candidates of interesting new types of objects that deserve more detailed domain-specific followup study. However, measurement data is nearly never free of measurement errors. These errors can generate false outliers that are not truly interesting. Although many approaches exist for finding outliers, they have no means to tell to what extent the peculiarity is not simply due to measurement errors. To address this issue, we have developed a model-based approach to infer genuine outliers from multivariate data sets when measurement error information is available. This is based on a probabilistic mixture of hierarchical density models, in which parameter estimation is made feasible by a tree-structured variational expectation-maximization algorithm. Here, we further develop an algorithmic enhancement to address the scalability of this approach, in order to make it applicable to large data sets, via a K-dimensional-tree based partitioning of the variational posterior assignments. This creates a non-trivial tradeoff between a more detailed noise model to enhance the detection accuracy, and the coarsened posterior representation to obtain computational speedup. Hence, we conduct extensive experimental validation to study the accuracy/speed tradeoffs achievable in a variety of data conditions. We find that, at low-to-moderate error levels, a speedup factor that is at least linear in the number of data points can be achieved without significantly sacrificing the detection accuracy. The benefits of including measurement error information into the modeling is evident in all situations, and the gain roughly recovers the loss incurred by the speedup procedure in large error conditions. We analyze and discuss in detail the characteristics of our algorithm based on results obtained on appropriately designed synthetic data experiments, and we also demonstrate its working in a real application example.


Acetylation reprograms MITF target selectivity and residence time.
The ability of transcription factors to discriminate between different classes of binding sites associated with specific biological functions underpins effective gene regulation in development and homeostasis. How this is achieved is poorly understood. The microphthalmia-associated transcription factor MITF is a lineage-survival oncogene that plays a crucial role in melanocyte development and melanoma. MITF suppresses invasion, reprograms metabolism and promotes both proliferation and differentiation. How MITF distinguishes between differentiation and proliferation-associated targets is unknown. Here we show that compared to many transcription factors MITF exhibits a very long residence time which is reduced by p300/CBP-mediated MITF acetylation at K206. While K206 acetylation also decreases genome-wide MITF DNA-binding affinity, it preferentially directs DNA binding away from differentiation-associated CATGTG motifs toward CACGTG elements. The results reveal an acetylation-mediated switch that suppresses differentiation and provides a mechanistic explanation of why a human K206Q MITF mutation is associated with Waardenburg syndrome.


Retinal gene expression after central retinal artery ligation: effects of ischemia and reperfusion.
To investigate the morphologic and molecular consequences of 30- and 90-minute central retinal artery ligation (CRAL)-induced retinal ischemia, followed by 3 and 12 hours of reperfusion, and to identify potential targets for therapy. Retinal ischemia was induced for 30 and 90 minutes by ligating the rat central retinal artery, and corresponding effects were examined histologically, immunocytochemically, and molecularly at 3 hours and 12 hours of reperfusion. Patterns of gene expression revealed significantly upregulated and downregulated genes by gene array analyses and were verified by quantitative RT-PCR. Functional pathways were correlated using gene set enrichment analysis. Substantial morphologic changes occurred from 3 hours to 7 days after CRAL in rats, resulting in a cellular loss in most retinal layers, particularly in inner nuclear and ganglion cell layers. After 30 minutes of CRAL and 3 hours of reperfusion, transcription-related genes such as ATF3, ID2, Klf4, BTG2, c-Fos, and c-Jun were activated. After 12 hours of reperfusion, the genes associated with kinase and caspase molecular pathways-including MAP kinases, Casp3 and Casp9-were upregulated. CRAL of 90 minutes and 3 hours of reperfusion induced glycolysis and gluconeogenesis-related genes such as G6PC. However, prolonged reperfusion of 12 hours was characterized by prominent activation of apoptosis-related genes, including Tp53, Akt1, Akt3, Pik3R1, Prkcb1, caspases (Casp3, Casp7, Casp9), and TNF. CRAL is a clinically relevant retinal ischemia model, and gene expression analysis can provide information regarding the molecular mechanisms underlying the pathophysiological processes during retinal ischemia. In addition, CRAL represents an effective experimental model with which to study retinal inflammation, development, aging, and, neurodegeneration.


Phagocytosis and intracellular fate of Aspergillus fumigatus conidia in alveolar macrophages.
Aspergillus fumigatus is the most prevalent airborne fungal pathogen responsible for fatal invasive aspergillosis in immunocompromised patients. Upon arrival in the lung alveolus, conidia of A. fumigatus are phagocytosed by alveolar macrophages, the major phagocytic cells of the lung. Engulfment and intracellular trafficking of A. fumigatus conidia in alveolar macrophages of two different origins, the murine cell line MH-S and human pulmonary alveolar macrophages, were analyzed by electron microscopy and immunofluorescence. Phagocytosis of A. fumigatus conidia required actin polymerization and phosphatidylinositol 3-kinase activity. Fusion of A. fumigatus phagosomes with early and late endosomes was shown by immunolabeling with specific markers for the transferrin receptor, early endosome antigen, and Rab7. Maturation of A. fumigatus phagolysosomes was monitored by using a fixable acidotropic probe, LysoTracker Red DND-99, and an anti-cathepsin D antibody. Bafilomycin A-induced inhibition of lysosomal acidification abolished the conidial killing by the macrophages. These data suggest that the maturation of A. fumigatus phagosomes results from fusion with the compartments of the endocytic pathway and that the killing of conidia depends on phagolysosome acidification. A model for the phagocytosis of A. fumigatus conidia by alveolar macrophages is proposed on the basis of these results.


Urban planning and health equity.
Although the fields of urban planning and public health share a common origin in the efforts of reformers to tame the ravages of early industrialization in the 19th century, the 2 disciplines parted ways in the early 20th century as planners increasingly focused on the built environment while public health professionals narrowed in on biomedical causes of disease and disability. Among the unfortunate results of this divergence was a tendency to discount the public health implications of planning decisions. Given increasingly complex urban environments and grave health disparities in cities worldwide, urban planners and public health professionals have once again become convinced of the need for inclusive approaches to improve population health and achieve health equity. To make substantive progress, intersectoral collaboration utilizing ecological and systems science perspectives will be crucial as the solutions lie well beyond the control of any single authority. Grounded in the social determinants of health, and with a renewed sense of interconnectedness, dedicated and talented people in government agencies and communities who recognize that our future depends on cultivating local change and evaluating the results can come to grips with the enormous challenge that lies ahead to create more equitable, sustainable, and healthier cities worldwide.


Expression of ENaC in LPS-induced inflammation of middle ear mucosa.
The expression of all three subunits of the epithelial sodium channel (ENaC) decreased after the lipopolysaccharide (LPS) injection and normalized as the fluid collection resolved. This implies that fluid collection in acute otitis media may be caused by the inhibition of ENaC function. We investigated the expression levels of subunits of ENaC after an injection of LPS into the middle ear cavity of rats. Seventy-two Sprague-Dawley rats were assigned randomly into the five groups that received an LPS injection and the control group. A transtympanic injection of LPS (1 mg/ml) was done and rats were sacrificed 6, 12, 24, 72, and 120 h after the procedure. Real-time RT-PCR was carried out for ENaC-α, -β, -γ, and immunohistochemistry was performed for ENaC-α and -β. The level of ENaC-α expression decreased 0.50-fold and 0.55-fold at 6 and 12 h, respectively (p < 0.05), but it normalized at 24 h. It increased 3.64-fold at 72 h and 3.24-fold at 120 h (p < 0.05). At 6 and 12 h after the LPS injection into the middle ear cavity, inflammation was induced and ENaC-α immunoreactivity decreased. At 24 h, ENaC-α immunoreactivity was normalized, then at 72 and 120 h after the injection it was increased.


Effects of grazing on leaf area index, fractional cover and evapotranspiration by a desert phreatophyte community at a former uranium mill site on the Colorado Plateau.
This study employed ground and remote sensing methods to monitor the effects of grazing on leaf area index (LAI), fractional cover (f(c)) and evapotranspiration (ET) of a desert phreatophyte community over an 11 year period at a former uranium mill site on the Colorado Plateau, U.S. Nitrate, ammonium and sulfate are migrating away from the mill site in a shallow alluvial aquifer. The phreatophyte community, consisting of Atriplex canescens (ATCA) and Sarcobatus vermiculatus (SAVE) shrubs, intercepts groundwater and could potentially slow the movement of the contaminant plume through evapotranspiration (ET). However, the site has been heavily grazed by livestock, reducing plant cover and LAI. We used livestock exclosures and revegetation plots to determine the effects of grazing on LAI, f(c) and ET, then projected the findings over the whole site using multi-platform remote sensing methods. We show that ET is approximately equal to annual precipitation at the site, but when ATCA and SAVE are protected from grazing they can develop high f(c) and LAI values, and ET can exceed annual precipitation, with the excess coming from groundwater discharge. Therefore, control of grazing could be an effective method to slow migration of contaminants at this and similar sites in the western U.S.


False accusations in an investigative context: differences between suggestible and non-suggestible witnesses.
False sexual abuse allegations have spurred research on suggestibility, on the assumption that leading questions may produce false accusations. Most researchers, however, have not measured the likelihood that those who respond to suggestive questioning will take the next step and make a formal (false) accusation. The present study incorporates both aspects of abuse investigations: suggestibility (i.e., responsiveness to questions in a leading interview) and false accusations (i.e., signing a formal complaint against an innocent suspect). Participants (N = 129) were observed in a laboratory session and then interviewed twice about their experiences by an interviewer who suggested that the laboratory assistant had behaved inappropriately. Although only 17% of the participants were suggestible, 39% agreed to sign the complaint. Suggestible participants were significantly more likely to make a false accusation than were non-suggestible participants. However, because of the low rate of suggestibility, most false accusations were made by non-suggestible participants. Implications for the legal system are discussed.


Using the BioAssay Ontology for analyzing high-throughput screening data.
High-throughput screening (HTS) is the main starting point for hit identification in drug discovery programs. This has led to a rapid increase of available screening data both within pharmaceutical companies and the public domain. We have used the BioAssay Ontology (BAO) 2.0 for assay annotation within AstraZeneca to enable comparison with external HTS methods. The annotated assays have been analyzed to identify technology gaps, evaluate new methods, verify active hits, and compare compound activity between in-house and PubChem assays. As an example, the binding of a fluorescent ligand to formyl peptide receptor 1 (FPR1, involved in inflammation, for example) in an in-house HTS was measured by fluorescence intensity. In total, 155 active compounds were also tested in an external ligand binding flow cytometry assay, a method not used for in-house HTS detection. Twelve percent of the 155 compounds were found active in both assays. By the annotation of assay protocols using BAO terms, internal and external assays can easily be identified and method comparison facilitated. They can be used to evaluate the effectiveness of different assay methods, design appropriate confirmatory and counterassays, and analyze the activity of compounds for identification of technology artifacts.


Does intra-aortic balloon support for myocardial infarction with cardiogenic shock improve outcome?
Thiele H, Zeymer U, Neumann FJ, Ferenc M, Olbrich HG, Hausleiter J, Richardt G, Hennersdorf M, Empen K, Fuernau G, Desch S, Eitel I, Hambrecht R, Fuhrmann J, Böhm M, Ebelt H, Schneider S, Schuler G, Werdan K; IABP-SHOCK II Trial Investigators: Intraaortic balloon support for myocardial infarction with cardiogenic shock. N Engl J Med 2012, 367:1287-1296. In the current international guidelines, intra-aortic balloon pump (IABP) counterpulsation is considered a class I treatment for acute myocardial infarction complicated by cardiogenic shock. However, evidence is based mainly on registry data, and there is a paucity of randomized clinical trials. To test the hypothesis that IABP counterpulsation, as compared with the best available medical therapy alone, results in a reduction in mortality among patients with acute myocardial infarction complicated by cardiogenic shock for whom early revascularization is planned. Randomized, prospective, open-label, multicenter trial. Thirty-seven centers in Germany. All adults had acute myocardial infarction complicated by cardiogenic shock and were expected to undergo early revascularization (by means of percutaneous coronary intervention or bypass surgery). After enrollment, 600 patients were randomly assigned to intra-aortic balloon counterpulsation (IABP group, 301 patients) or no IABP counterpulsation (control group, 299 patients). The primary efficacy endpoint is 30-day all-cause mortality. At 30 days, 119 patients in the IABP group (39.7%) and 123 patients in the control group (41.3%) had died (relative risk with IABP, 0.96; 95% confidence interval, 0.79 to 1.17; P=0.69). There were no significant differences in secondary endpoints or in process-of-care measures, including the time to hemodynamic stabilization, the length of stay in the intensive care unit, serum lactate levels, the dose and duration of catecholamine therapy, and renal function. The use of IABP counterpulsation did not significantly reduce 30-day mortality in patients with acute myocardial infarction complicated by cardiogenic shock for whom an early revascularization strategy was planned.


Theoretical description of effective heat transfer between two viscously coupled beads.
We analytically study the role of nonconservative forces, namely viscous couplings, on the statistical properties of the energy flux between two Brownian particles kept at different temperatures. From the dynamical model describing the system, we identify an energy flow that satisfies a fluctuation theorem both in the stationary and in transient states. In particular, for the specific case of a linear nonconservative interaction, we derive an exact fluctuation theorem that holds for any measurement time in the transient regime, and which involves the energy flux alone. Moreover, in this regime the system presents an interesting asymmetry between the hot and cold particles. The theoretical predictions are in good agreement with the experimental results already presented in our previous article [Imparato et al., Phys. Rev. Lett. 116, 068301 (2016)PRLTAO0031-900710.1103/PhysRevLett.116.068301], where we investigated the thermodynamic properties of two Brownian particles, trapped with optical tweezers, interacting through a dissipative hydrodynamic coupling.


The effect of select pulp cavity conditions on stress field development in distal abutments in two types of fixed dental prostheses.
Insufficient coronal tooth structure may require restoration of endodontically treated (ET) teeth with cast posts and cores (CPCs). The prognosis for these teeth is a matter of scientific debate, especially if they serve as distal abutments in cantilever fixed dental prostheses (FDPs). The purpose of this study was to study stress field development in distal abutments in two types of FDPs with different pulp cavity conditions. The methodology involved the development of four digital models in which the right mandibular premolars were splinted via an FDP with: (1) no cantilever and a vital distal abutment, (2) no cantilever and an ET CPC distal abutment, (3) a single-unit cantilever and a vital distal abutment, and (4) a single-unit cantilever with an ET CPC distal abutment. The models were analyzed using a three-dimensional finite element program, and von Mises stress values and patterns were evaluated. The results revealed that although the stress distribution patterns in dentin were dissimilar, the von Mises stress values registered for the vital and ET CPC distal abutment were not considerably different. However, higher stress values were detected in the dentin area surrounding the post-gutta-percha interface after CPC placement. The addition of the cantilever resulted in a considerable increase in stress on the dental tissue structures. CPCs appear to create a risk of potential fracture that is initiated in the dentin at the apex of the post. The type of restoration appears to have a much more serious impact on the stress pattern developed in the distal abutment, and the addition of a cantilever appears to biomechanically compromise both biologic and restorative structures.