Validating the TeleStroke Mimic Score: A Prediction Rule for Identifying Stroke Mimics Evaluated Over Telestroke Networks.

BACKGROUND AND PURPOSE: Up to 30% of acute stroke evaluations are deemed stroke mimics, and these are common in telestroke as well. We recently published a risk prediction score for use during telestroke encounters to differentiate stroke mimics from ischemic cerebrovascular disease derived and validated in the Partners TeleStroke Network. Using data from 3 distinct US and European telestroke networks, we sought to externally validate the TeleStroke Mimic (TM) score in a broader population. METHODS: We evaluated the TM score in 1930 telestroke consults from the University of Utah, Georgia Regents University, and the German TeleMedical Project for Integrative Stroke Care Network. We report the area under the curve in receiver-operating characteristic curve analysis with 95% confidence interval for our previously derived TM score in which lower TM scores correspond with a higher likelihood of being a stroke mimic. RESULTS: Based on final diagnosis at the end of the telestroke consultation, there were 630 of 1930 (32.6%) stroke mimics in the external validation cohort. All 6 variables included in the score were significantly different between patients with ischemic cerebrovascular disease versus stroke mimics. The TM score performed well (area under curve, 0.72; 95% confidence interval, 0.70-0.73; P<0.001), similar to our prior external validation in the Partners National Telestroke Network. CONCLUSIONS: The TM score's ability to predict the presence of a stroke mimic during telestroke consultation in these diverse cohorts was similar to its performance in our original cohort. Predictive decision-support tools like the TM score may help highlight key clinical differences between mimics and patients with stroke during complex, time-critical telestroke evaluations.

Linear and nonlinear genetic relationships between type traits and productive life in US dairy goats.

Linear or nonlinear genetic relationships between productive life and functional productive life at 72 mo, with final score (SCO), stature, strength, dairyness (DAI), teat diameter, rear legs (side view), rump angle, rump width (RUW), fore udder attachment (FUA), rear udder height, rear udder arch, udder depth (UDD), suspensory ligament (SUS), and teat placement, as well as heritabilities and correlations were estimated from multibreed US dairy goat records. Productive life was defined as the total days in production until 72 mo of age (PL72) for goats having the opportunity to express the trait. Functional productive life (FPL72) was analyzed by incorporating first lactation milk yield, fat yield, protein yield, and SCO in the statistical model. Heritabilities and correlations were estimated using linear mixed models with pedigree additive genetic relationships and ASReml software. Nonlinearity of genetic relationships was assessed based on second-degree polynomial (quadratic) regression models, with the breeding values of PL72 or FPL72 as responses and the breeding values for each type trait (linear and quadratic) as predictor variables. Heritability estimates were 0.19, 0.14, 0.18, 0.20, 0.14, 0.07, 0.28, 0.20, 0.15, 0.13, 0.25, 0.18, 0.20, 0.21, 0.21, and 0.32 for PL72, FPL72, SCO, stature, strength, DAI, teat diameter, rear legs, rump angle, RUW, FUA, rear udder height, rear udder arch, UDD, SUS, and teat placement, respectively. The type traits SCO, RUW, and FUA were the most correlated with PL72 and FPL72, so these may be used as selection criteria to increase longevity in dairy goats. An increase in the coefficient of determination >1% for the second degree, compared with that for the linear model for either PL72 or FPL72, was taken as evidence of a nonlinear genetic relationship. Using this criterion, PL72 showed maximum values at intermediate scores in DAI, UDD, and RUW, and maximum values at extreme scores in FUA and SUS, whereas FPL72 showed maximum values at intermediate scores in DAI and UDD, and maximum values at extreme scores in FUA, RUW, and SUS. Selecting for increased SCO, RUW, and FUA will lead to an increase of FPL72 in goats. Consideration of nonlinear relationships between DAI, FUA, RUW, SUS, and UDD may help in the design of more efficient breeding programs for dairy goats using conformation traits.

Estimation of genetic parameters for productive life, reproduction, and milk-production traits in US dairy goats.

Heritabilities and correlations for milk yield (MY), fat yield (FY), protein yield (PY), combined fat and protein yield (FPY), fat percentage (F%), protein percentage (P%), age at first kidding (AFK), interval between the first and second kidding (KI), and real and functional productive life at 72mo (FPL72) of 33,725 US dairy goats, were estimated using animal models. Productive life was defined as the total days in production until 72mo of age (PL72) for goats having the opportunity to express the trait. Functional productive life was obtained by correcting PL72 for MY, FY, PY, and final type score (FS). Six selection indexes were used, including or excluding PL72, with 6 groups of different economic weights, to estimate the responses to selection considering MY, FY, PY, and PL72 as selection criteria. The main criteria that determined the culling of a goat from the herd were low FS, MY, and FY per lactation. Heritability estimates were 0.22, 0.17, 0.37, 0.37, 0.38, 0.39, 0.54, 0.64, 0.09, and 0.16 for PL72, FPL72, MY, FY, PY, FPY, F%, P%, KI, and AFK, respectively. Most genetic correlations between the evaluated traits and PL72 or FPL72 were positive, except for F% (-0.04 and -0.06, respectively), P% (-0.002 and -0.03, respectively), and AFK (-0.03 and -0.01, respectively). The highest genetic correlations were between FPL72 and MY (0.39) and between PL72 and MY (0.33). Most phenotypic correlations between the traits evaluated and FPL72 and PL72 were positive (>0.23 and >0.26, respectively), except for F% (-0.004 and -0.02, respectively), P% (-0.05 and -0.02), KI (-0.01 and -0.07), and AFK (-0.08 and -0.08). The direct selection for PL72 increased it by 102.28d per generation. The use of MY, FY, PY, KI, or AFK as selection criteria increased PL72 by 39.21, 27.33, 35.90, -8.28, or 2.77d per generation, respectively. The inclusion of PL72 as selection criterion increased the expected response per generation from 0.15 to 17.35% in all selection indices studied.

Relating sensory and chemical properties of sour cream to consumer acceptance.

Sour cream is a widely popular acidified dairy product. Volatile compounds and organic acids and their specific contributions to flavor or acceptance have not been established, nor has a comprehensive study been conducted to characterize drivers of liking for sour cream. The objective of this study was to characterize chemical and sensory properties of sour cream and to determine the drivers of liking for sour cream. Descriptive sensory and instrumental analyses followed by consumer testing were conducted. Flavor and texture attributes of 32 (22 full-fat, 6 reduced-fat, and 4 fat-free) commercial sour creams were evaluated by a trained descriptive sensory panel. Percent solids, percent fat, pH, titratable acidity, and colorimetric measurements were conducted to characterize physical properties of sour creams. Organic acids were evaluated by HPLC and volatile aroma active compounds were evaluated by gas chromatography-mass spectrometry with gas chromatography-olfactometry. Consumer acceptance testing (n=201) was conducted on selected sour creams, followed by external preference mapping. Full-fat sour creams were characterized by the lack of surface gloss and chalky textural attributes, whereas reduced-fat and fat-free samples displayed high intensities of these attributes. Full-fat sour creams were higher in cooked/milky and milk fat flavors than the reduced-fat and fat-free samples. Reduced-fat and fat-free sour creams were characterized by cardboard, acetaldehyde/green, and potato flavors, bitter taste, and astringency. Lactic acid was the prominent organic acid in all sour creams, followed by acetic and citric acids. High aroma-impact volatile compounds in sour creams were 2,3-butanedione, acetic acid, butyric acid, octanal, 2-methyl-3-furanthiol, 1-octene-3-one, and acetaldehyde. Positive drivers of liking for sour cream were milk fat, cooked/milky and sweet aromatic flavors, opacity, color intensity, and adhesiveness. This comprehensive study established sensory and instrumental properties of sour creams and their relationship to consumer acceptance.

The impact of polygamy on women's mental health: a systematic review.

Aims. The purpose of this systematic review is to identify and appraise the present state of prevalence research on the mental health of polygynous women, or plural wives, and to summarize its implications for future research and social work practice. Methods. PsycInfo (1967 to November 2011) and Medline (1985 to November 2011) databases, systematic bibliography hand-searches, personal communication with a leading expert, and gray literature searching were applied in a systematic literature search of the prevalence of mental-health issues in polygynous women compared to monogamous women. Twenty-two studies meeting eligibility criteria were identified. Study characteristics, methods and findings were systematically extracted and appraised for quality. Results. The identified studies are of mixed methodological quality, but generally suggest a more significant prevalence of mental-health issues in polygynous women compared to monogamous women. Individual studies report a higher prevalence of somatization, depression, anxiety, hostility, psychoticism and psychiatric disorder in polygynous wives as well as reduced life and marital satisfaction, problematic family functioning and low self-esteem. Conclusions. The current state of the research reveals with moderate confidence, a more significant prevalence of mental-health issues in polygynous women as compared to monogamous women. Implications for practice and research are indicated.

Sensory properties and drivers of liking for Greek yogurts.

Greek yogurt is currently the largest growing sector in the dairy industry. Because no standard of identity exists for Greek yogurts in the United States, and they can be made by a variety of methods, variability in sensory properties is expected. Knowledge of consumer perception and specific drivers of liking will be useful information for product developers. The objective of this study was to document the sensory properties of commercial Greek yogurts and to determine drivers of liking through descriptive profiling and consumer testing. Flavor and texture attributes of commercial Greek yogurts (n = 24) were evaluated in triplicate by a trained descriptive sensory panel. An online survey (n = 520) was used to collect consumer usage and attitude information for Greek yogurts before consumer acceptance testing. Consumer acceptance testing (n = 155) was then conducted on commercial Greek yogurts (n = 10). Univariate and multivariate statistical analyses were used for data analysis. Sensory properties of yogurt differed with fat content and manufacture (Greek vs. fortified Greek). Full-fat yogurts were characterized by firmness and denseness, whereas low- and non-fat yogurts lacked firmness, denseness, cohesiveness, and, after stirring, viscosity. Fortified Greek yogurts generally had more surface shine and jiggle and lower denseness compared with traditional Greek yogurts. Fewer flavor differences were observed among yogurts compared with texture differences. Fortified Greek yogurts displayed a burnt/beefy flavor not documented in traditional Greek yogurts, but this flavor was not evident in all fortified Greek yogurts. Consumer preferred Greek yogurts with firm, dense texture, moderate sweet aromatic, milkfat and dairy sour flavors, and moderate sour taste. Consumers were aware of the increased protein content of Greek yogurts but generally unaware of differences between strained and fortified Greek yogurts; both strained Greek and fortified Greek yogurts received the highest overall liking scores in blinded acceptance testing. Successful Greek yogurts can be manufactured using addition of dried dairy ingredients or by traditional straining and centrifugation.

Breed differences over time and heritability estimates for production and reproduction traits of dairy goats in the United States.

To aid in improvement of breeding programs for production and reproduction traits of US dairy goats, breed differences over time were documented and genetic parameters were estimated. Data were from herds with ≥2 breeds (Alpine, LaMancha, Nubian, Oberhasli, Saanen, or Toggenburg), but only purebred data were analyzed. Three kidding periods were examined: 1976 through 1984, 1985 through 1994, and 1995 through 2005. Univariate repeatability mixed models were used to estimate least squares means by kidding period-breed and genetic parameters for milk, fat, and protein yields, combined fat and protein yield, fat and protein percentages, protein:fat ratio, age at first kidding, and kidding interval. Trends across kidding periods were favorable for most yield traits for all breeds but generally unfavorable for reproduction traits. Saanens had the highest milk (1,063 to 1,125 kg) and protein yields (31 to 33 kg). Nubians had the highest fat yields (37 to 40 kg) and lowest milk yields (791 to 851 kg). Oberhaslis had the lowest fat (31 to 33 kg) and protein (23 to 27 kg) yields. Alpines had the largest increase in milk yield (7.4%); Oberhaslis had the largest increase in protein (17.4%) and combined fat and protein (13.2%) yields. Combined fat and protein yield was higher for Nubians, Saanens, and Alpines (65 to 72 kg) than for LaManchas, Toggenburgs, and Oberhaslis (53 to 67 kg). Nubians had the highest fat (4.7 to 4.8%) and protein (3.6 to 3.8%) percentages. Only Nubians increased in fat percentage (2.1%); protein percentage increased most for Toggenburgs (7.4%) and Alpines (7.1%). Protein:fat ratio was highest for Toggenburgs (0.84 to 0.89) and lowest for Nubians (0.76 to 0.81), but Nubians had the largest increase in protein:fat ratio (6.6%). Saanens were oldest at first kidding (509 to 589 d), and Toggenburgs and LaManchas generally were youngest (435 to 545 d); age at first kidding increased most for Alpines (21.8%) and LaManchas (21.6%). Kidding intervals generally were shorter for Oberhaslis, LaManchas, and Nubians (350 to 377 d) than for Toggenburgs, Alpines, and Saanens (373 to 387 d). Kidding interval increased most for Nubians (3.9%) and Saanens (3.8%) and decreased only for Oberhaslis (5.4%). Heritability estimates across breeds were 0.35 for milk and fat yields, 0.37 for protein yield and protein:fat ratio, 0.36 for combined fat and protein yield, 0.52 for fat percentage, 0.54 for protein percentage, 0.23 for age at first kidding, and 0.05 for kidding interval. Genetic selection within breed is feasible for production and reproduction traits of US dairy goats.

Short communication: genetic and environmental relationships between milk yield and kidding interval in dairy goats.

First-parity 305-d milk yield and intervals between first and second kiddings from 1975 through 2005 were analyzed to estimate genetic and environmental parameters for United States Alpine, LaMancha, Nubian, Saanen, and Toggenburg dairy goats. The data set included 43,612 does with first-parity milk yield and 25,863 does with first kidding interval. Parameters were estimated by REML using bivariate mixed models within and across breeds. Fixed effects were month and herd-year of kidding within breed and month and breed-herd-year of kidding across breeds. Random effects were animal and residual. The heritability estimate for first-parity milk yield was 0.36+/-0.01 across breeds and ranged from 0.35 to 0.38 within breed; heritability for first kidding interval was 0.05+/-0.01 across breeds and ranged from 0.00 to 0.15 within breed. The estimate of the genetic correlation between first-parity milk yield and first kidding interval was positive (unfavorable) across breeds (0.35+/-0.09). Estimates of environmental correlations within and across breeds were positive (0.16-0.25). The presence of unfavorable genetic relationships between milk yield and kidding interval indicates a need to include reproductive performance as a selection criterion.

A bioassessment approach for mid-continent great rivers: the Upper Mississippi, Missouri, and Ohio (USA).

The objectives of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE) are to (1) develop and demonstrate, in collaboration with states, an assessment program yielding spatially unbiased estimates of the condition of mid-continent great rivers; (2) evaluate environmental indicators for assessing great rivers; and (3) assess the current condition of selected great river resources. The purpose of this paper is to describe EMAP-GRE using examples based on data collected in 2004-2006 with emphasis on an approach to determining reference conditions. EMAP-GRE includes the Upper Mississippi River, the Missouri River, and the Ohio River. Indicators include biotic assemblages (fish, macroinvertebrates, plankton, algae), water chemistry, and aquatic and riparian physical habitat. Reference strata (river reaches for which a single reference expectation is appropriate) were determined by ordination of the fish assemblage and examination of spatial variation in environmental variables. Least disturbed condition of fish assemblages for reference strata was determined by empirical modeling in which we related fish assemblage metrics to a multimetric stressor gradient. We inferred least disturbed condition from the y-intercept, the predicted condition when stress was least. Thresholds for dividing the resource into management-relevant condition classes for biotic indicators were derived using predicted least disturbed condition to set the upper bound on the least disturbed condition class. Also discussed are the outputs of EMAP-GRE, including the assessment document, multimetric indices of condition, and unbiased data supporting state and tribal Clean Water Act reporting, adaptive management, and river restoration.

HGS-ETR1, a fully human TRAIL-receptor 1 monoclonal antibody, induces cell death in multiple tumour types in vitro and in vivo.

Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in a variety of tumour cells through activation of TRAIL-R1 and TRAIL-R2 death signalling receptors. Here, we describe the characterisation and activity of HGS-ETR1, the first fully human, agonistic TRAIL-R1 mAb that is being developed as an antitumour therapeutic agent. HGS-ETR1 showed specific binding to TRAIL-R1 receptor. HGS-ETR1 reduced the viability of multiple types of tumour cells in vitro, and induced activation of caspase 8, Bid, caspase 9, caspase 3, and cleavage of PARP, indicating activation of TRAIL-R1 alone was sufficient to induce both extrinsic and intrinsic apoptotic pathways. Treatment of cell lines in vitro with HGS-ETR1 enhanced the cytotoxicity of chemotherapeutic agents (camptothecin, cisplatin, carboplatin, or 5-fluorouracil) even in tumour cell lines that were not sensitive to HGS-ETR1 alone. In vivo administration of HGS-ETR1 resulted in rapid tumour regression or repression of tumour growth in pre-established colon, non-small-cell lung, and renal tumours in xenograft models. Combination of HGS-ETR1 with chemotherapeutic agents (topotecan, 5-fluorouracil, and irinotecan) in three independent colon cancer xenograft models resulted in an enhanced antitumour efficacy compared to either agent alone. Pharmacokinetic studies in the mouse following intravenous injection showed that HGS-ETR1 serum concentrations were biphasic with a terminal half-life of 6.9-8.7 days and a steady-state volume of distribution of approximately 60 ml kg(-1). Clearance was 3.6-5.7 ml(-1) day(-1) kg(-1). These data suggest that HGS-ETR1 is a specific and potent antitumour agent with favourable pharmacokinetic characteristics and the potential to provide therapeutic benefit for a broad range of human malignancies.

Effect of dietary protein on lean body wasting in dogs: correlation between loss of lean mass and markers of proteasome-dependent proteolysis.

To determine the effect of dietary protein intake on lean body wasting in adult canines a study was undertaken to investigate the Ubiquitin Proteasome (UP) pathway and concurrent changes in lean and fat body mass of canines fed variable sources and concentrations of dietary protein. Purpose-bred, intact female canines (56) between the ages of 2 and 3 years were fed either 12 or 28% protein diet for 10 weeks. Each diet contained variable amounts of corn gluten meal and chicken protein sources in ratios of 100 : 0, 67 : 33, 33 : 67 and 0 : 100 per cent (w/w), respectively. All diets were isocaloric with calories coming from protein : fat : carbohydrate at the respective ratios of 12 : 40 : 48% for the 12% diets, and 28 : 40 : 32% for the 28% diets. Standard dual energy X-ray absorptiometry was performed to assess total body lean and fat mass at weeks 0 and 10 of the dietary trial. Muscle biopsies were also taken and processed for protein determination and standard gel electrophoresis with subsequent Western blotting for 20S proteasome and PA700 regulatory cap subunit p31. Statistical analysis revealed a moderate degree of correlation between increasing quantities of corn gluten, which is low in essential amino acids (i.e. lysine, tryptophan), and increasing loss of lean body mass over the 10-week study (R = 0.56). Furthermore, a moderate degree of correlation was observed between increasing concentrations of corn gluten protein and decreased expression of the p31 subunit of the 26S proteasome (R = 0.49). Additionally, the dogs consuming the 12% protein diets had a significant increase in fat mass regardless of the protein source. These findings suggest that lean body wasting in adult canines can be associated with the consumption of low protein diets consisting of predominantly corn gluten, which is likely due to imbalances or subclinical deficiencies of specific essential amino acids, and that low protein diets may augment accumulation of adipose tissue. Although the mechanisms remain unclear, alteration of molecular targets of skeletal muscle proteolysis, specifically involving the UP pathway occur.

The use of DNA array technology in studies of ocular viral pathogenesis.

DNA arrays can be used to simultaneously analyze the expression of hundreds of genes and permit systematic approaches to biological discovery with a potentially profound impact on genomics, pharmacogenomics, and proteomics. Microarrays have been used to study host-pathogen interactions, and recently this technology has been applied to investigate host-virus interactions. DNA arrays are used to monitor host alterations in several virus-induced cancers and upon infection with wild-type or modified viruses, or viral gene products. Alternatively, viral chips are used to characterize the transcriptional program of pathogenic viruses and in antiviral drug screening and drug resistance. With an aim to extend the use of this technology to ocular research, and specifically to study changes in host cell transcription in ocular adenovirus infection, we used a commercial array to compare adenovirus-infected human corneal cells to mock-infected cells. Of the 1176 genes analyzed, 72 genes associated with cell cycle regulation, apoptosis, oncogenesis, transcription, signaling, and inflammation were differentially regulated. In this review we summarize the use of DNA arrays in the study of viral infections and suggest potential uses of the technology in ocular viral pathogenesis research.

Extracellular superoxide production by Enterococcus faecalis requires demethylmenaquinone and is attenuated by functional terminal quinol oxidases.

The intestinal commensal bacterium, Enterococcus faecalis, is unusual among prokaryotic organisms in its ability to produce substantial extracellular superoxide. Transposon mutagenesis, allelic replacement, and electron spin resonance (ESR)-spin trapping showed that superoxide production and generation of derivative hydroxyl radical were dependent on membrane-associated demethylmenaquinone. Extracellular superoxide was generated through univalent reduction of oxygen by reduced demethylmenaquinone. Moreover, extracellular superoxide production was inhibited by exogenous haematin, an essential cofactor for cytochrome bd, and by fumarate, a substrate for fumarate reductase. As integral membrane quinol oxidases, cytochrome bd and fumarate reductase redox cycle demethylmenaquinone, and are necessary for aerobic and anaerobic respiration respectively. A rat model of intestinal colonization demonstrated that conditions exist in the mammalian intestinal tract that permit a mode of respiration for E. faecalis that results in the formation of hydroxyl radical. These results identify and characterize the mechanism by which E. faecalis generates extracellular free radicals.

Constitutive activation of NF-kappa B and secretion of interleukin-8 induced by the G protein-coupled receptor of Kaposi's sarcoma-associated herpesvirus involve G alpha(13) and RhoA.

The Kaposi's sarcoma herpesvirus (KSHV) open reading frame 74 encodes a G protein-coupled receptor (GPCR) for chemokines. Exogenous expression of this constitutively active GPCR leads to cell transformation and vascular overgrowth characteristic of Kaposi's sarcoma. We show here that expression of KSHV-GPCR in transfected cells results in constitutive transactivation of nuclear factor kappa B (NF-kappa B) and secretion of interleukin-8, and this response involves activation of G alpha(13) and RhoA. The induced expression of a NF-kappa B luciferase reporter was partially reduced by pertussis toxin and the G beta gamma scavenger transducin, and enhanced by co-expression of G alpha(13) and to a lesser extent, G alpha(q). These results indicate coupling of KSHV-GPCR to multiple G proteins for NF-kappa B activation. Expression of KSHV-GPCR led to stress fiber formation in NIH 3T3 cells. To examine the involvement of the G alpha(13)-RhoA pathway in KSHV-GPCR-mediated NF-kappa B activation, HeLa cells were transfected with KSHV-GPCR alone and in combination with the regulator of G protein signaling (RGS) from p115RhoGEF or a dominant negative RhoA(T19N). Both constructs, as well as the C3 exoenzyme from Clostritium botulinum, partially reduced NF-kappa B activation by KSHV-GPCR, and by a constitutively active G alpha(13)(Q226L). KSHV-GPCR-induced NF-kappa B activation is accompanied by increased secretion of IL-8, a function mimicked by the activated G alpha(13) but not by an activated G alpha(q)(Q209L). These results suggest coupling of KSHV-GPCR to the G alpha(13)-RhoA pathway in addition to other G proteins.

The mechanism of pore assembly for a cholesterol-dependent cytolysin: formation of a large prepore complex precedes the insertion of the transmembrane beta-hairpins.

Perfringolysin O (PFO) is a member of the cholesterol-dependent cytolysin (CDC) family of membrane-penetrating toxins. The CDCs form large homooligomers (estimated to be comprised of up to 50 CDC monomers) that are responsible for generating a large pore in cholesterol-containing membranes of eukaryotic cells. The assembly of the PFO cytolytic complex was examined to determine whether it forms an oligomeric prepore complex on the membrane prior to the insertion of its membrane-spanning beta-sheet. A PFO oligomeric complex was formed on liposomes at both 4 degrees C and 37 degrees C and shown by SDS-agarose gel electrophoresis to be comprised of a large, comparatively homogeneous complex instead of a distribution of oligomer sizes. At low temperature, the processes of oligomerization and membrane insertion could be resolved, and PFO was found to form an oligomer without significant membrane insertion of its beta-hairpins. Furthermore, PFO was found to increase the ion conductivity through a planar bilayer by large and discrete stepwise changes in conductance that are consistent with the insertion of a preassembled pore complex into the bilayer. The combined results of these analyses strongly support the hypothesis that PFO forms a large oligomeric prepore complex on the membrane surface prior to the insertion of its transmembrane beta-sheet.

The mechanism of membrane insertion for a cholesterol-dependent cytolysin: a novel paradigm for pore-forming toxins.

Perfringolysin O (PFO), a water-soluble monomeric cytolysin secreted by pathogenic Clostridium perfringens, oligomerizes and forms large pores upon encountering cholesterol-containing membranes. Whereas all pore-forming bacterial toxins examined previously have been shown to penetrate the membrane using a single amphipathic beta hairpin per polypeptide, cysteine-scanning mutagenesis and multiple independent fluorescence techniques here reveal that each PFO monomer contains a second domain involved in pore formation, and that each of the two amphipathic beta hairpins completely spans the membrane. In the soluble monomer, these transmembrane segments are folded into six alpha helices. The insertion of two transmembrane hairpins per toxin monomer and the major change in secondary structure are striking and define a novel paradigm for the mechanism of membrane insertion by a cytolytic toxin.

Surgical management of rectal and jejunal tears associated with dystocia in a cow.

The rectal and jejunal tears associated with dystocia in a cow were treated surgically. The cow developed peritonitis which was treated medically. It recovered, was successfully rebred and delivered a live calf.

Identification of a membrane-spanning domain of the thiol-activated pore-forming toxin Clostridium perfringens perfringolysin O: an alpha-helical to beta-sheet transition identified by fluorescence spectroscopy.

Clostridium perfringens perfringolysin O (PFO or theta-toxin) is a cytolytic toxin that binds to cholesterol-containing membranes and then self-associates to spontaneously form aqueous pores of varying size in the bilayer. In this study, a membrane-spanning domain has been identified in PFO by a combination of fluorescence spectroscopic methods using the fluorescent dye N, N'-dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1, 3-diazolyl)ethylenediamine (NBD) whose emission properties are sensitive to water. PFO was substituted with a single cysteine at most of the residues between amino acids K189 and N218, and then each cysteine was modified with NBD. Each purified NBD-labeled PFO was then bound to membranes, and the probe's environment was ascertained by measuring its fluorescence lifetime, emission intensity, and collisional quenching with either aqueous (iodide ions) or nonaqueous (nitroxide-labeled phospholipids) quenchers. Lifetime and intensity measurements revealed that the amino acid side chains in this region of the membrane-bound PFO polypeptide alternated between being in an aqueous or a nonaqueous environment. This pattern indicates that this portion of the membrane-bound PFO spans the membrane in an antiparallel beta-sheet conformation. The alternating exposure of these residues to the hydrophobic interior of the bilayer was demonstrated by their susceptibility to quenching by nitroxide moieties attached to phospholipid acyl chains. Residues K189-N218 therefore form a two-stranded, amphipathic beta-sheet in the membrane-bound PFO that creates a stable interface between the pore and the membrane. This same region packs as three short alpha-helices in the soluble, monomeric form of PFO, and therefore, the cholesterol-dependent conversion of PFO to a membrane-bound oligomer involves a major structural transition in which three alpha-helices unfold to form a membrane-spanning amphipathic beta-sheet.

Sustained attention and impulsivity in children with Tourette syndrome: comorbidity and confounds.

We sought to determine whether Tourette syndrome (TS) was related to attentional disturbance independent of Attention Deficit Hyperactivity disorder (ADHD) and obsessive-compulsive symptoms (OCS), two comorbid conditions associated with attentional problems in other populations. Participants were 74 children [21 with uncomplicated TS (TS-Only), 14 with TS and ADHD (TS-ADHD), 21 with ADHD, and 18 controls]. Impaired sustained attention and impulsivity on a Continuous Performance Task were uncommon in the TS-Only group, but common in TS-ADHD and ADHD. The TS-ADHD group was less impulsive than the ADHD group, but had a higher rate of whole-body tics than the TS-Only group, raising questions about the diagnosis of ADHD in TS. OCS was not an independent risk factor for attention problems. However, the association of tic severity and impulsivity was consistent with the theory that TS involves dysfunction in cortical-subcortical circuits mediating behavioral inhibition.