Weight maintenance as a tight rope walk - a Grounded Theory study.
Overweight and obesity are considerable public health problems internationally as well as in Sweden. The long-term results of obesity treatment are modest as reported by other studies. The importance of extending the focus to not only comprise obesity treatment but also prevention of weight gain is therefore being emphasized. However, despite the suggested change in focus there is still no consensus on how to prevent obesity or maintain weight. This study reports findings from a qualitative study focusing on attitudes, behaviors and strategies important for primary weight maintenance in a middle-aged population. In depth interviews were conducted with 23 maintainers and four slight gainers in Sweden. The interviews were transcribed and an analysis of weight maintenance was performed using Grounded Theory. Based on the informants' stories, describing attitudes, behaviors and strategies of importance for primary weight maintenance, a model illustrating the main findings, was constructed. Weight maintenance was seen as "a tightrope walk" and four strategies of significance for this "tightrope walk" were described as "to rely on heritage", "to find the joy", "to find the routine" and "to be in control". Eleven "ideal types" were included in the model to illustrate different ways of relating to the main strategies. These "ideal types" described more specific attitudes and behaviors such as; eating food that is both tasteful and nutritious, and choosing exercise that provides joy. However, other somewhat contradictory behaviors were also found such as; only eating nutritious food regardless of taste, and being physically active to control stress and emotions. This study show great variety with regards to attitudes, strategies and behaviors important for weight maintenance, and considerations need to be taken before putting the model into practice. However, the results from this study can be used within primary health care by enhancing the understanding of how people differ in their relation to food and physical activity. It informs health personnel about the need to differentiate advices related to body weight, not only to different sub-groups of individuals aiming at losing weight but also to sub-groups of primary weight maintainers aiming at maintaining weight.


On the role of ice-nucleating aerosol in the formation of ice particles in tropical mesoscale convective systems.
Over decades, the cloud physics community has debated the nature and role of aerosol particles in ice initiation. The present study shows that the measured concentration of ice crystals in tropical mesoscale convective systems exceeds the concentration of ice nucleating particles (INPs) by several orders of magnitude. The concentration of INPs was assessed from the measured aerosol particles concentration in the size range of 0.5 to 1 µm. The observations from this study suggest that primary ice crystals formed on INPs make only a minor contribution to the total concentration of ice crystals in tropical mesoscale convective systems. This is found by comparing the predicted INP number concentrations with in-situ ice particle number concentrations. The obtained measurements suggest that ice multiplication is the likely explanation for the observed high concentrations of ice crystals in this type of convective system.


Large-scale bi-level strain design approaches and mixed-integer programming solution techniques.
The use of computational models in metabolic engineering has been increasing as more genome-scale metabolic models and computational approaches become available. Various computational approaches have been developed to predict how genetic perturbations affect metabolic behavior at a systems level, and have been successfully used to engineer microbial strains with improved primary or secondary metabolite production. However, identification of metabolic engineering strategies involving a large number of perturbations is currently limited by computational resources due to the size of genome-scale models and the combinatorial nature of the problem. In this study, we present (i) two new bi-level strain design approaches using mixed-integer programming (MIP), and (ii) general solution techniques that improve the performance of MIP-based bi-level approaches. The first approach (SimOptStrain) simultaneously considers gene deletion and non-native reaction addition, while the second approach (BiMOMA) uses minimization of metabolic adjustment to predict knockout behavior in a MIP-based bi-level problem for the first time. Our general MIP solution techniques significantly reduced the CPU times needed to find optimal strategies when applied to an existing strain design approach (OptORF) (e.g., from ∼10 days to ∼5 minutes for metabolic engineering strategies with 4 gene deletions), and identified strategies for producing compounds where previous studies could not (e.g., malate and serine). Additionally, we found novel strategies using SimOptStrain with higher predicted production levels (for succinate and glycerol) than could have been found using an existing approach that considers network additions and deletions in sequential steps rather than simultaneously. Finally, using BiMOMA we found novel strategies involving large numbers of modifications (for pyruvate and glutamate), which sequential search and genetic algorithms were unable to find. The approaches and solution techniques developed here will facilitate the strain design process and extend the scope of its application to metabolic engineering.


Insulin resistance and risk of ischemic stroke among nondiabetic individuals from the northern Manhattan study.
Whether insulin resistance predicts ischemic stroke (IS) is still a matter of debate. To determine the association between insulin resistance (IR) and risk of first ischemic stroke in a large, multiethnic, stroke-free cohort without diabetes. Prospective, population-based cohort study. Longitudinal epidemiologic study. A cohort of 1509 nondiabetic participants from the Northern Manhattan Study (mean [SD] age, 11 [10] years; 64.2% women; 58.9% Hispanics). Insulin sensitivity, expressed by the homeostasis model assessment (HOMA) of insulin sensitivity (HOMA index = [fasting insulin × fasting glucose] / 22.5). Insulin resistance was defined by a HOMA-IR index in the top quartile (Q4). Cox proportional hazards models were used to determine the effect of HOMA-IR on the risk of incident IS, myocardial infarction (MI), vascular death, and combined outcomes (IS, MI, and vascular death). The mean (SD) HOMA-IR was 2.3 (2.1), and Q4 was at least 2.8. During mean follow-up of 8.5 years, vascular events occurred in 180 participants; 46 had fatal or nonfatal IS, 45 had fatal or nonfatal MI, and 121 died of vascular causes. The HOMA-IR Q4 vs less than Q4 significantly predicted the risk of IS only (adjusted hazard ratio, 2.83; 95% confidence interval, 1.34-5.99) but not other vascular events. This effect was independent of sex, race/ethnicity, traditional vascular risk factors, and metabolic syndrome and its components. Insulin resistance estimated using the HOMA is a marker of increased risk of incident stroke in nondiabetic individuals. These findings emphasize the need to better characterize individuals at increased risk for IS and the potential role of primary preventive therapies targeted at IR.


The Impact of Peer Substance Use and Polygenic Risk on Trajectories of Heavy Episodic Drinking Across Adolescence and Emerging Adulthood.
Heavy episodic drinking is developmentally normative among adolescents and young adults, but is linked to adverse consequences in later life, such as drug and alcohol dependence. Genetic and peer influences are robust predictors of heavy episodic drinking in youth, but little is known about the interplay between polygenic risk and peer influences as they impact developmental patterns of heavy episodic drinking. Data were from a multisite prospective study of alcohol use among adolescents and young adults with genome-wide association data (n = 412). Generalized linear mixed models were used to characterize the initial status and slopes of heavy episodic drinking between age 15 and 28. Polygenic risk scores (PRS) were derived from a separate genome-wide association study for alcohol dependence and examined for their interaction with substance use among the adolescents' closest friends in predicting the initial status and slopes of heavy episodic drinking. Close friend substance use was a robust predictor of adolescent heavy episodic drinking, even after controlling for parental knowledge and peer substance use in the school. PRS were predictive of the initial status and early patterns of heavy episodic drinking in males, but not in females. No interaction was detected between PRS and close friend substance use for heavy episodic drinking trajectories in either males or females. Although substance use among close friends and genetic influences play an important role in predicting heavy episodic drinking trajectories, particularly during the late adolescent to early adult years, we found no evidence of interaction between these influences after controlling for other social processes, such as parental knowledge and broader substance use among other peers outside of close friends. The use of longitudinal models and accounting for multiple social influences may be crucial for future studies focused on uncovering gene-environment interplay. Clinical implications are also discussed.


[Serum sialic acid, cellular anionic charge and erythrocyte aggregation in diabetic and hypertensive patients].
The aim of this work was to study the relationship between serum sialic acid (SSA) and erythrocyte anionic charge (EAC) with erythrocyte aggregation in two groups: diabetic (DBT, n=20) and hypertensive (HT, n=21) patients, compared to a control group (n=20). We worked with anticoagulated blood with EDTA and serum. The erythrocyte aggregation was studied by microscopically observing and quantifying aggregates using an ASP (Aggregate Shape Parameter). The EAC was determined by binding an Alcian blue dye to the membrane sialic acid and SSA was determined by spectrophotometric method with an Erlich reactant. The values of ASP and SSA increased significantly in HT and DBT patients compared to the control group. The HT and DBT groups showed amorphous aggregates, evident in an alteration in the values of ASP, which were significantly higher ( p < 0.005) than in healthy patients. The EAC values were much lower in HT and DBT patients than in the control group (p < 0.0001). In this work, abnormalities in the erythrocyte aggregation could be detected by the values of ASP, EAC and SSA, which might be involved in vascular disorders of diseases such as hypertension and diabetes.


The genome sequence of the spontaneously hypertensive rat: Analysis and functional significance.
The spontaneously hypertensive rat (SHR) is the most widely studied animal model of hypertension. Scores of SHR quantitative loci (QTLs) have been mapped for hypertension and other phenotypes. We have sequenced the SHR/OlaIpcv genome at 10.7-fold coverage by paired-end sequencing on the Illumina platform. We identified 3.6 million high-quality single nucleotide polymorphisms (SNPs) between the SHR/OlaIpcv and Brown Norway (BN) reference genome, with a high rate of validation (sensitivity 96.3%-98.0% and specificity 99%-100%). We also identified 343,243 short indels between the SHR/OlaIpcv and reference genomes. These SNPs and indels resulted in 161 gain or loss of stop codons and 629 frameshifts compared with the BN reference sequence. We also identified 13,438 larger deletions that result in complete or partial absence of 107 genes in the SHR/OlaIpcv genome compared with the BN reference and 588 copy number variants (CNVs) that overlap with the gene regions of 688 genes. Genomic regions containing genes whose expression had been previously mapped as cis-regulated expression quantitative trait loci (eQTLs) were significantly enriched with SNPs, short indels, and larger deletions, suggesting that some of these variants have functional effects on gene expression. Genes that were affected by major alterations in their coding sequence were highly enriched for genes related to ion transport, transport, and plasma membrane localization, providing insights into the likely molecular and cellular basis of hypertension and other phenotypes specific to the SHR strain. This near complete catalog of genomic differences between two extensively studied rat strains provides the starting point for complete elucidation, at the molecular level, of the physiological and pathophysiological phenotypic differences between individuals from these strains.


Role of Language-Related Functional Connectivity in Patients with Benign Childhood Epilepsy with Centrotemporal Spikes.
Benign childhood epilepsy with centrotemporal spikes (BECTS) does not always have a benign cognitive outcome. We investigated the relationship between cognitive performance and altered functional connectivity (FC) in the resting-state brain networks of BECTS patients. We studied 42 subjects, comprising 19 BECTS patients and 23 healthy controls. Cognitive performance was assessed using the Korean version of the Wechsler Intelligence Scale for Children-III, in addition to verbal and visuospatial memory tests and executive function tests. Resting-state functional magnetic resonance imaging was acquired in addition to high-resolution structural data. We selected Rolandic and language-related areas as regions of interest (ROIs) and analyzed the seed-based FC to voxels throughout the brain. We evaluated the correlations between the neuropsychological test scores and seed-based FC values using the same ROIs. The verbal intelligence quotient (VIQ) and full-scale intelligence quotient (FSIQ) were lower in BECTS patients than in healthy controls (p<0.001). The prevalence of subjects with a higher performance IQ than VIQ was significantly higher in BECTS patients than in healthy controls (73.7% vs. 26.1%, respectively; p=0.002). Both the Rolandic and language-related ROIs exhibited more enhanced FC to voxels in the left inferior temporal gyrus in BECTS patients than in healthy controls. A particularly interestingly finding was that the enhanced FC was correlated with lower cognitive performance as measured by the VIQ and the FSIQ in both patients and control subjects. Our findings suggest that the FC alterations in resting-state brain networks related to the seizure onset zone and language processing areas could be related to adaptive plasticity for coping with cognitive dysfunction.


Clinical Characteristics of Pregnant Women on the Use of Daily Low-dose Aspirin in Different Hypertensive Pregnancy Disorders: A Retrospective Comparative Study.
Hypertensive disorders represent major causes of maternal and fetal complications. It includes a range of conditions, most notably preeclampsia. Aspirin is a well-accepted therapy for the primary and secondary prevention of cardiovascular events. The indications for the use of aspirin during pregnancy are, however, the subject of much concern. This study aimed to assess the clinical characteristics from the benefits of daily low-dose aspirin administration alongside antihypertensive in pregnant women with different hypertensive disorders. A retrospective observational study was carried out on pregnant women during their routine visit to the obstetric clinic at Baghdad Teaching Hospital. Patients diagnosed during pregnancy with different hypertensive disorders on the prescription of antihypertensive medication with or without daily low-dose aspirin administration were selected. Data were collected to structure a detailed assessment regarding the patients' demographic, gestational, and medical records. Methyldopa was the main antihypertensive agent (98%). Among pregnant women with daily aspirin use, 68% had gestational hypertension, 24% had preeclampsia alongside proteinuria (P = 0.0001), the frequency of daily dose intake of methyldopa (250 mg) tablet (two vs. three times) was significant (P = 0.0001). All pregnant women within the group of daily low-dose aspirin were safe from the incidence of eclampsia (P = 0.0001). This study provides intriguing evidence for the benefits of daily low-dose aspirin use during pregnancy as it is considered as a simple protective measure from serious maternal complications of hypertensive disorders, where these complications continue to affect maternal health long after delivery.


Studies on motility and fertility of cooled stallion spermatozoa.
This study on extended, cooled stallion spermatozoa aimed to compare the ability of three extenders to maintain sperm motility during 24 h of preservation, and to describe pregnancy and foaling rates after artificial insemination (AI) of stallion spermatozoa stored and transported in the extender chosen from the in vitro study. After 6 and 24 h of preservation, motility, both subjective and evaluated by the motility analyzer (total, progressive and rapid), was lower in non-fat, dried skim milk-glucose than in both other extenders: dried skim milk-glucose added to 2% centrifuged egg yolk, and ultra high temperature treated skim milk-sugar-saline solution added to 2% centrifuged egg yolk (INRA82-Y). Rapid spermatozoa and sperm velocity parameters, after 24 h, were significantly higher in INRA82-Y. In the fertility trial, semen collected from three Maremmano stallions, diluted in INRA82-Y, and transported in a refrigerated Styrofoam box, was used to inseminate 56 mares of the same breed. Pregnancy rates after the first cycle and per breeding season were significantly higher for the 31 mares inseminated in three AI centres (54.8 and 80.6%, respectively) than for the 25 mares inseminated at the breeder's facilities (28.0 and 52.0%). Foaling rates were not significantly different between the AI centres mares (54.8%) and the other mares (44.0%). In conclusion, INRA82-Y yielded satisfactory pregnancy and foaling rates, especially when employed in the more controlled situation of an AI centre, and can therefore be included among those available for cooled stallion semen preservation.


Electrical and optical properties of an organic semiconductor based on polyaniline prepared by emulsion polymerization and fabrication of Ag/polyaniline/n-Si Schottky diode.
The electrical, optical, and metal-semiconductor contact properties of the polyaniline prepared by emulsion polymerization have been investigated to obtain an organic semiconductor material. The obtained results suggest that the polyaniline (PANI) studied is an organic semiconductor material with optical band gap (E(g) = 2.21 eV) and room electrical conductivity (sigma(25) = 3.12 x 10(-2) S/cm) values. A Schottky diode with configuration Ag/PANI/n-Si was fabricated. The ideality factor and barrier height of Ag/PANI/n-Si diode at room temperature were found to be 4.59 and 0.38 eV, respectively. The obtained diode parameters change with temperature. The Richardson constant A* value for the Ag/PANI/n-Si diode was found to be 3.81 x 10(-4) A/cm(2).K. The Ag/PANI/n-Si diode is a metal-insulator-semiconductor-type device. The standard deviation, which is a measure of the barrier homogeneity, was found to be 0.14, indicating the presence of interface inhomogeneities. It can be concluded that the polyaniline prepared by emulsion polymerization is an organic semiconductor and Ag/PANI/n-Si configuration shows a Schottky contact.


Loss of Nocturnin, a circadian deadenylase, confers resistance to hepatic steatosis and diet-induced obesity.
The mammalian circadian system consists of a central oscillator in the suprachiasmatic nucleus of the hypothalamus, which coordinates peripheral clocks in organs throughout the body. Although circadian clocks control the rhythmic expression of a large number of genes involved in metabolism and other aspects of circadian physiology, the consequences of genetic disruption of circadian-controlled pathways remain poorly defined. Here we report that the targeted disruption of Nocturnin (Ccrn4l) in mice, a gene that encodes a circadian deadenylase, confers resistance to diet-induced obesity. Mice lacking Nocturnin remain lean on high-fat diets, with lower body weight and reduced visceral fat. However, unlike lean lipodystrophic mouse models, these mice do not have fatty livers and do not exhibit increased activity or reduced food intake. Gene expression data suggest that Nocturnin knockout mice have deficits in lipid metabolism or uptake, in addition to changes in glucose and insulin sensitivity. Our data support a pivotal role for Nocturnin downstream of the circadian clockwork in the posttranscriptional regulation of genes necessary for nutrient uptake, metabolism, and storage.


Mathematical modeling identifies optimum lapatinib dosing schedules for the treatment of glioblastoma patients.
Human primary glioblastomas (GBM) often harbor mutations within the epidermal growth factor receptor (EGFR). Treatment of EGFR-mutant GBM cell lines with the EGFR/HER2 tyrosine kinase inhibitor lapatinib can effectively induce cell death in these models. However, EGFR inhibitors have shown little efficacy in the clinic, partly because of inappropriate dosing. Here, we developed a computational approach to model the in vitro cellular dynamics of the EGFR-mutant cell line SF268 in response to different lapatinib concentrations and dosing schedules. We then used this approach to identify an effective treatment strategy within the clinical toxicity limits of lapatinib, and developed a partial differential equation modeling approach to study the in vivo GBM treatment response by taking into account the heterogeneous and diffusive nature of the disease. Despite the inability of lapatinib to induce tumor regressions with a continuous daily schedule, our modeling approach consistently predicts that continuous dosing remains the best clinically feasible strategy for slowing down tumor growth and lowering overall tumor burden, compared to pulsatile schedules currently known to be tolerated, even when considering drug resistance, reduced lapatinib tumor concentrations due to the blood brain barrier, and the phenotypic switch from proliferative to migratory cell phenotypes that occurs in hypoxic microenvironments. Our mathematical modeling and statistical analysis platform provides a rational method for comparing treatment schedules in search for optimal dosing strategies for glioblastoma and other cancer types.


Associated features in females with an FMR1 premutation.
Changes in the fragile X mental retardation 1 gene (FMR1) have been associated with specific phenotypes, most specifically those of fragile X syndrome (FXS), fragile X tremor/ataxia syndrome (FXTAS), and fragile X primary ovarian insufficiency (FXPOI). Evidence of increased risk for additional medical, psychiatric, and cognitive features and conditions is now known to exist for individuals with a premutation, although some features have been more thoroughly studied than others. This review highlights the literature on medical, reproductive, cognitive, and psychiatric features, primarily in females, that have been suggested to be associated with changes in the FMR1 gene. Based on this review, each feature is evaluated with regard to the strength of evidence of association with the premutation. Areas of need for additional focused research and possible intervention strategies are suggested.


Identification of land use with water quality data in stormwater using a neural network.
To control stormwater pollution effectively, development of innovative, land-use-related control strategies will be required. An approach that could differentiate land-use types from stormwater quality would be the first step to solving this problem. We propose a neural network approach to examine the relationship between stormwater water quality and various types of land use. The neural network model can be used to identify land-use types for future known and unknown cases. The neural model uses a Bayesian network and has 10 water quality input variables, four neurons in the hidden layer, and five land-use target variables (commercial, industrial, residential, transportation, and vacant). We obtained 92.3 percent of correct classification and 0.157 root-mean-squared error on test files. Based on the neural model, simulations were performed to predict the land-use type of a known data set, which was not used when developing the model. The simulation accurately described the behavior of the new data set. This study demonstrates that a neural network can be effectively used to produce land-use type classification with water quality data.


The Hebrew version of the FibroFatigue scale: validation of a questionnaire for assessment of fibromyalgia and chronic fatigue syndrome.
The objective of this study is to validate a translated Hebrew version of the FibroFatigue Scale (FFS). The Hebrew version of the FFS was administered to 100 patients fulfilling ACR criteria for classification of FM together with the validated Hebrew version of the Fibromyalgia Impact Questionnaire (FIQ), the validated Hebrew version of the Short Form-36 (SF-36) and a Visual Analogue Scale (VAS) measurement of pain, anxiety, depression, morning stiffness and global well being. Test-retest reliability was assessed using Spearman correlations. Internal consistency was evaluated with Cronbach's alpha of reliability. Construct validity of the FFS was evaluated by correlations among the FFS, the FIQ and the subscales of the SF-36. Mean duration of symptoms was 10.7 years, and mean age of participants was 53.5 years. Test-retest reliability was between 0.46 and 0.85 for the various FFS items. Internal consistency was 0.89 for the overall FFS. Significant correlations were obtained between the FFS items and the SF-36. These results support the reliability and validity of the data obtained with the Hebrew version of the FSS for detecting and measuring symptom severity in Hebrew speaking patients with FM.


Contributions of yap1 Mutation and Subsequent atrF Upregulation to Voriconazole Resistance in Aspergillus flavus.
Aspergillus flavus is the second most significant pathogenic cause of invasive aspergillosis; however, its emergence risks and mechanisms of voriconazole (VRC) resistance have not yet been elucidated in detail. Here, we demonstrate that repeated exposure of A. flavus to subinhibitory concentrations of VRC in vitro causes the emergence of a VRC-resistant mutant with a novel resistance mechanism. The VRC-resistant mutant shows a MIC of 16 μg/ml for VRC and of 0.5 μg/ml for itraconazole (ITC). Whole-genome sequencing analysis showed that the mutant possesses a point mutation in yap1, which encodes a bZIP transcription factor working as the master regulator of the oxidative stress response, but no mutations in the cyp51 genes. This point mutation in yap1 caused alteration of Leu558 to Trp (Yap1Leu558Trp) in the putative nuclear export sequence in the carboxy-terminal cysteine-rich domain of Yap1. This Yap1Leu558Trp substitution was confirmed as being responsible for the VRC-resistant phenotype, but not for that of ITC, by the revertant to Yap1wild type with homologous gene replacement. Furthermore, Yap1Leu558Trp caused marked upregulation of the atrF ATP-binding cassette transporter, and the deletion of atrF restored susceptibility to VRC in A. flavus These findings provide new insights into VRC resistance mechanisms via a transcriptional factor mutation that is independent of the cyp51 gene mutation in A. flavus.


Life-stage factors associated with overweight severity in adolescents.
Investigating life-stage factors associated with overweight may be useful in the prevention of excessive BMI increase. The main aim of this study was to investigate the influence of the route of delivery, birth weight and overweight onset on overweight severity in a sample of overweight adolescents followed at a Pediatric Obesity Clinic. Clinical data from 412 adolescents with overweight (BMI ≥ p85), aged 10-18 were retrospectively collected and analyzed. Adolescents born by cesarean section (CS) showed a lower age of overweight onset, compared to other methods of delivery (d= 0.33, p= .009). Birth weight was positively associated with BMI z-score (r=.164, p= 002) and waist circumference (WC) (r=.191, p=.001). The overweight onset was negatively associated with BMI z-score (r= -.277, p < .001), WC (r= -.270, p < .001) and body fat mass (r= -.199, p=.001). Overweight duration was the best predictor of BMI z-score, explaining in 75% its variation (F=1,317)=26.94, p < .001), which increased to 99% when birth weight was included in the model (F(2,316)=18.47, p < .001). This study suggests that lifestyle may interrupt the burden of CS on BMI z-score throughout growth. Moreover, increased birth weight may anticipate overweight onset, and consequently overweight duration in the presence of inadequate lifestyle behaviors.


A Roadmap Toward the Definition of Actionable Tumor-Specific Antigens.
The search for tumor-specific antigens (TSAs) has considerably accelerated during the past decade due to the improvement of proteogenomic detection methods. This provides new opportunities for the development of novel antitumoral immunotherapies to mount an efficient T cell response against one or multiple types of tumors. While the identification of mutated antigens originating from coding exons has provided relatively few TSA candidates, the possibility of enlarging the repertoire of targetable TSAs by looking at antigens arising from non-canonical open reading frames opens up interesting avenues for cancer immunotherapy. In this review, we outline the potential sources of TSAs and the mechanisms responsible for their expression strictly in cancer cells. In line with the heterogeneity of cancer, we propose that discrete families of TSAs may be enriched in specific cancer types.


Bacterial Leaf Spot of Radicchio (Cichorium intybus) is caused by Xanthomonas hortorum.
Radicchio (Cichorium intybus) is ranked 22 among crops in Monterey County, California, with a farm gate value of $19,531,000 (3). Beginning in 2002, a leaf spot disease of radicchio was observed in Monterey County. The disease began as small lesions and in some cases coalesced into larger, irregular spots. Lesions were maroon to dark brown; in some cases, the margins of brown lesions became dark maroon with aging. Each leaf spot was observable from both adaxial and abaxial leaf surfaces. Symptoms primarily occurred on the outer foliage of the heads, though on occasion the head cap leaf could develop lesions. Disease incidence in the first year resulted in up to 10% unharvested radicchio because of cap leaf infections or reduced head size if outer wrapper leaves were all removed; outbreaks in subsequent seasons were more limited. Bacteria forming yellow mucoid colonies were isolated from surface disinfested symptomatic tissue that was macerated and streaked onto sucrose peptone agar medium. Bacteria were gram negative, did not fluoresce on King's Medium B, and used esculin as a carbon source but used none of the other 48 carbon sources tested using the API 50 CH test strip. Nine isolates from symptomatic radicchio had the same DNA fragment banding pattern generated by repetitive extragenic palindromic sequence polymerase chain reactions (rep-PCR) using the BOXA1R primer. Amplicons of rpoD, dnaK, fyuA, and gyrB for multilocus sequence typing (MLST) were generated using a modification of the scheme developed by Young et al. (4) and sequenced by a commercial laboratory. Concatenated sequences of the four genes from the radicchio isolates were compared to the sequences available in the Plant Associated and Environmental Microbes Database (1). The genetic distance between the nine isolates from radicchio and pathotypes of Xanthomonas hortorum were 0.03 or less and MLST analysis indicated that radicchio isolates were members of the species X. hortorum (2). To complete Koch's postulates, freshly grown cultures were suspended in phosphate buffer and adjusted to approximately 5 × 108 CFU/ml. The inoculum was sprayed onto the undersides of leaves of 40-day-old radicchio plants (C. intybus cv. Leonardo). Plants were incubated at 100% humidity for 48 h and then moved to a greenhouse. Plants sprayed with buffer served as negative controls. For each of the two experiments conducted, there were three and six single-plant replicates per treatment. The buffer treated plants did not develop symptoms but the plants treated with isolates from radicchio developed leaf spots similar to those observed in the field with symptoms beginning to be visible after 5 days. The bacteria isolated from symptomatic tissue on inoculated plants were identical to the original strains when compared with rep-PCR, thus completing Koch's postulates. Results from the two experiments were similar. To our knowledge, this is the first report of X. hortorum causing a leaf spot disease on radicchio. The disease continues to occur sporadically on radicchio grown in coastal California. References: (1) Almeida et al. Phytopathology 100:208, 2010. (2) Bull et al. Phytopathology 101:847, 2011. (3) Lauritzen, Monterey County Crop Report, 2010; (4) Young et al. Syst. Appl. Microbiol. 31:366, 2008.


Near-Field Spectroscopy of Cylindrical Phonon-Polariton Antennas.
Surface phonon polaritons (SPhPs) are hybrid light-matter states in which light strongly couples to lattice vibrations inside the Reststrahlen band of polar dielectrics at mid-infrared frequencies. Antennas supporting localized surface phonon polaritons (LSPhPs) easily outperform their plasmonic counterparts operating in the visible or near-infrared in terms of field enhancement and confinement thanks to the inherently slower phonon-phonon scattering processes governing SPhP decay. In particular, LSPhP antennas have attracted considerable interest for thermal management at the nanoscale, where the emission strongly diverts from the usual far-field blackbody radiation due to the presence of evanescent waves at the surface. However, far-field measurements cannot shed light on the behavior of antennas in the near-field region. To overcome this limitation, we employ scattering-scanning near-field optical microscopy (sSNOM) to unveil the spectral near-field response of 3C-SiC antenna arrays. We present a detailed description of the behavior of the antenna resonances by comparing far-field and near-field spectra and demonstrate the existence of a mode with no net dipole moment, absent in the far-field spectra, but of importance for applications that exploit the heightened electromagnetic near fields. Furthermore, we investigate the perturbation in the antenna response induced by the presence of the AFM tip, which can be further extended toward situations where for example strong IR emitters couple to LSPhP modes.


Prognosis of Cryptogenic Stroke With Patent Foramen Ovale at Older Ages and Implications for Trials: A Population-Based Study and Systematic Review.
Patent foramen ovale (PFO) closure may prevent recurrent stroke after cryptogenic transient ischemic attack (TIA) or stroke (TIA/stroke) in patients aged 60 years or younger. Patent foramen ovale is associated with cryptogenic stroke in the older population, but risk of recurrence is unknown. Data on prognosis of patients receiving medical treatment at older ages (≥60 years) are essential to justify trials of PFO closure. To examine the age-specific risk of recurrence in patients with cryptogenic TIA/stroke with PFO. A prospective study was nested in the population-based Oxford Vascular Study between September 1, 2014, and March 31, 2019, with face-to-face follow-up for 5 years. A total of 416 consecutive patients with a diagnosis of cryptogenic TIA or nondisabling stroke, screened for PFO at a rapid-access TIA/stroke clinic, were included. A systematic review and meta-analysis of cohort studies reporting on ischemic stroke recurrence after cryptogenic TIA/stroke in patients with PFO who were receiving medical therapy alone, or with PFO vs no-PFO was conducted. Sample size calculation for future trials on PFO closure was performed for patients aged 60 years or older. Patent foramen ovale and age as modifiers of risk of recurrent stroke after cryptogenic TIA/stroke in patients receiving only medical therapy. Risk of ischemic stroke recurrence in patients with cryptogenic TIA/stroke and PFO receiving medical therapy only, and in patients with vs without PFO, stratified by age (<65 vs ≥65 years), as well as sample-size calculation for future trials of PFO closure in patients aged 60 years or older. Among the 153 Oxford Vascular Study patients with PFO (mean [SD] age, 66.7 [13.7] years; 80 [52.3%] men), recurrent ischemic stroke risk (2.05 per 100 patient-years) was similar to the pooled estimate from a systematic review of 23 other studies (9 trials and 14 observational studies) (2.00 per 100 patient-years; 95% CI, 1.55-2.58). However, there was heterogeneity between studies (P < .001 for heterogeneity), owing mainly to risk increasing with mean cohort age (meta-regression: R2 = 0.31; P = .003). In the pooled analysis of 4 studies including patients with or without PFO, increased risk of stroke recurrence with PFO was seen only at age 65 years or older (odds ratio, 2.5; 95% CI, 1.4-4.2; P = .001 for difference; P = .39 for heterogeneity). The pooled ischemic stroke risk was 3.27 per 100 patient-years (95% CI, 2.59-4.13) in 4 cohorts with mean age 60 years or older. Assuming the more conservative 2.0 per 100 patient-years ischemic stroke risk in the PFO nonclosure arms of future trials in patients aged 60 years or older, projected sample sizes were 1080 per arm for 80% power to detect a 33% relative risk reduction. The findings of this study suggest that age is a determinant of risk of ischemic stroke after cryptogenic TIA/stroke in patients with PFO, such that trials of PFO closure at older ages are justified; however, projected sample sizes are large.


Positive imagery cognitive bias modification (CBM) and internet-based cognitive behavioral therapy (iCBT): a randomized controlled trial.
Accruing evidence suggests that positive imagery-based cognitive bias modification (CBM) could have potential as a standalone targeted intervention for depressive symptoms or as an adjunct to existing treatments. We sought to establish the benefit of this form of CBM when delivered prior to Internet cognitive behavioral therapy (iCBT) for depression A randomized controlled trial (RCT) of a 1-week Internet-delivered positive CBM vs. an active control condition for participants (N=75, 69% female, mean age=42) meeting diagnostic criteria for major depression; followed by a 10-week iCBT program for both groups. Modified intent-to-treat marginal and mixed effect models demonstrated no significant difference between conditions following the CBM intervention or the iCBT program. In both conditions there were significant reductions (Cohen׳s d .57-1.58, 95% CI=.12-2.07) in primary measures of depression and interpretation bias (PHQ9, BDI-II, AST-D). Large effect size reductions (Cohen׳s d .81-1.32, 95% CI=.31-1.79) were observed for secondary measures of distress, disability, anxiety and repetitive negative thinking (K10, WHODAS, STAI, RTQ). Per protocol analyses conducted in the sample of participants who completed all seven sessions of CBM indicated between-group superiority of the positive over control group on depression symptoms (PHQ9, BDI-II) and psychological distress (K10) following CBM (Hedges g .55-.88, 95% CI=-.03-1.46) and following iCBT (PHQ9, K10). The majority (>70%) no longer met diagnostic criteria for depression at 3-month follow-up. The control condition contained many active components and therefore may have represented a smaller 'dose' of the positive condition. Results provide preliminary support for the successful integration of imagery-based CBM into an existing Internet-based treatment for depression.


Tailored delivery of analgesic ziconotide across a blood brain barrier model using viral nanocontainers.
The blood brain barrier (BBB) is often an insurmountable obstacle for a large number of candidate drugs, including peptides, antibiotics, and chemotherapeutic agents. Devising an adroit delivery method to cross the BBB is essential to unlocking widespread application of peptide therapeutics. Presented here is an engineered nanocontainer for delivering peptidic drugs across the BBB encapsulating the analgesic marine snail peptide ziconotide (Prialt®). We developed a bi-functional viral nanocontainer based on the Salmonella typhimurium bacteriophage P22 capsid, genetically incorporating ziconotide in the interior cavity, and chemically attaching cell penetrating HIV-Tat peptide on the exterior of the capsid. Virus like particles (VLPs) of P22 containing ziconotide were successfully transported in several BBB models of rat and human brain microvascular endothelial cells (BMVEC) using a recyclable noncytotoxic endocytic pathway. This work demonstrates proof in principle for developing a possible alternative to intrathecal injection of ziconotide using a tunable VLP drug delivery nanocontainer to cross the BBB.


Investigation of the thermal and photochemical reactions of ozone with 2,3-dimethyl-2-butene.
The matrix isolation technique, combined with infrared spectroscopy and twin jet codeposition, has been used to characterize intermediates formed during the ozonolysis of 2,3-dimethyl-2-butene (DMB). Absorptions of early intermediates in the twin jet experiments grew up to 200% upon annealing to 35 K. A number of these absorptions have been assigned to the elusive Criegee intermediate (CI) and secondary ozonide (SOZ) of DMB, transient species not previously observed for this system. Also observed was the primary ozonide (POZ), in agreement with earlier studies. The wavelength dependence of the photodestruction of these product bands was explored with irradiation from λ ≥ 220 to ≥580 nm. Merged jet (flow reactor) experiments generated "late" stable oxidation products of DMB. A recently developed concentric jet method was also utilized to increase yields and monitor the concentration of intermediates and products formed at different times by varying the length of mixing distance (d = 0 to -11 cm) before reaching the cold cell for spectroscopic detection. Identification of intermediates formed during the ozonolysis of DMB was further supported by (18)O and scrambled (16,18)O isotopic labeling experiments as well as theoretical density functional calculations at the B3LYP/6-311++G(d,2p) level.


Cross-Milieu Terrorist Collaboration: Using Game Theory to Assess the Risk of a Novel Threat.
This article uses a game-theoretic approach to analyze the risk of cross-milieu terrorist collaboration-the possibility that, despite marked ideological differences, extremist groups from very different milieus might align to a degree where operational collaboration against Western societies becomes possible. Based upon theoretical insights drawn from a variety of literatures, a bargaining model is constructed that reflects the various benefits and costs for terrorists' collaboration across ideological milieus. Analyzed in both sequential and simultaneous decision-making contexts and through numerical simulations, the model confirms several theoretical arguments. The most important of these is that although likely to be quite rare, successful collaboration across terrorist milieus is indeed feasible in certain circumstances. The model also highlights several structural elements that might play a larger role than previously recognized in the collaboration decision, including that the prospect of nonmaterial gains (amplification of terror and reputational boost) plays at least as important a role in the decision to collaborate as potential increased capabilities does. Numerical simulation further suggests that prospects for successful collaboration over most scenarios (including operational) increase when a large, effective Islamist terrorist organization initiates collaboration with a smaller right-wing group, as compared with the other scenarios considered. Although the small number of historical cases precludes robust statistical validation, the simulation results are supported by existing empirical evidence of collaboration between Islamists and right- or left-wing extremists. The game-theoretic approach, therefore, provides guidance regarding the circumstances under which such an unholy alliance of violent actors is likely to succeed.


Identification and analysis of genome-wide SNPs provide insight into signatures of selection and domestication in channel catfish (Ictalurus punctatus).
Domestication and selection for important performance traits can impact the genome, which is most often reflected by reduced heterozygosity in and surrounding genes related to traits affected by selection. In this study, analysis of the genomic impact caused by domestication and artificial selection was conducted by investigating the signatures of selection using single nucleotide polymorphisms (SNPs) in channel catfish (Ictalurus punctatus). A total of 8.4 million candidate SNPs were identified by using next generation sequencing. On average, the channel catfish genome harbors one SNP per 116 bp. Approximately 6.6 million, 5.3 million, 4.9 million, 7.1 million and 6.7 million SNPs were detected in the Marion, Thompson, USDA103, Hatchery strain, and wild population, respectively. The allele frequencies of 407,861 SNPs differed significantly between the domestic and wild populations. With these SNPs, 23 genomic regions with putative selective sweeps were identified that included 11 genes. Although the function for the majority of the genes remain unknown in catfish, several genes with known function related to aquaculture performance traits were included in the regions with selective sweeps. These included hypoxia-inducible factor 1β. HIFιβ.. and the transporter gene ATP-binding cassette sub-family B member 5 (ABCB5). HIF1β. is important for response to hypoxia and tolerance to low oxygen levels is a critical aquaculture trait. The large numbers of SNPs identified from this study are valuable for the development of high-density SNP arrays for genetic and genomic studies of performance traits in catfish.


Production and Comprehension of Prosodic Markers in Sign Language Imperatives.
In signed and spoken language sentences, imperative mood and the corresponding speech acts such as for instance, command, permission or advice, can be distinguished by morphosyntactic structures, but also solely by prosodic cues, which are the focus of this paper. These cues can express paralinguistic mental states or grammatical meaning, and we show that in American Sign Language (ASL), they also exhibit the function, scope, and alignment of prosodic, linguistic elements of sign languages. The production and comprehension of prosodic facial expressions and temporal patterns therefore can shed light on how cues are grammaticalized in sign languages. They can also be informative about the formal semantic and pragmatic properties of imperative types not only in ASL, but also more broadly. This paper includes three studies: one of production (Study 1) and two of comprehension (Studies 2 and 3). In Study 1, six prosodic cues are analyzed in production: temporal cues of sign and hold duration, and non-manual cues including tilts of the head, head nods, widening of the eyes, and presence of mouthings. Results of Study 1 show that neutral sentences and commands are well distinguished from each other and from other imperative speech acts via these prosodic cues alone; there is more limited differentiation among explanation, permission, and advice. The comprehension of these five speech acts is investigated in Deaf ASL signers in Study 2, and in three additional groups in Study 3: Deaf signers of German Sign Language (DGS), hearing non-signers from the United States, and hearing non-signers from Germany. Results of Studies 2 and 3 show that the ASL group performs significantly better than the other 3 groups and that all groups perform above chance for all meaning types in comprehension. Language-specific knowledge, therefore, has a significant effect on identifying imperatives based on targeted cues. Command has the most cues associated with it and is the most accurately identified imperative type across groups-indicating, we suggest, its special status as the strongest imperative in terms of addressing the speaker's goals. Our findings support the view that the cues are accessible in their content across groups, but that their language-particular combinatorial possibilities and distribution within sentences provide an advantage to ASL signers in comprehension and attest to their prosodic status.


A DFT calculation-inspired Rh(i)-catalyzed reaction via suppression of α-H shift in α-alkyldiazoacetates.
Metal-associated carbenes from diazo compounds promote many useful chemistry transformations in modern organic chemistry. However, compared to α-aryldiazoacetate-derived carbenes (ArDCs), the synthetic application of α-alkyldiazoacetate-derived carbenes (AlDCs) is greatly limited due to intramolecular α-H transfer (elimination) that results in alkenes as the main by-products. An intriguing α-alkyldiazoacetate-involved three-component reaction has been developed following DFT calculation inspiration to provide β-hydroxyl α-alkyl-α-amino acid derivatives in good yields. The intramolecular α-H shift of an α-alkyldiazoacetate-derived carbene was successfully suppressed by the association of a Rh(i) complex to form the corresponding active ammonium ylide, which was trapped before the fast 1,2-H transfer process. A Rh(i)-chiral diene complex was identified as an effective catalyst to give an asymmetric version of the reaction with good enantioselectivity. This reaction provides insight into extending the efficient transformation of α-alkyldiazoacetate-derived carbenes and their synthetic application.


Lifespan and Healthspan: Past, Present, and Promise.
The past century was a period of increasing life expectancy throughout the age range. This resulted in more people living to old age and to spending more years at the older ages. It is likely that increases in life expectancy at older ages will continue, but life expectancy at birth is unlikely to reach levels above 95 unless there is a fundamental change in our ability to delay the aging process. We have yet to experience much compression of morbidity as the age of onset of most health problems has not increased markedly. In recent decades, there have been some reductions in the prevalence of physical disability and dementia. At the same time, the prevalence of disease has increased markedly, in large part due to treatment which extends life for those with disease. Compressing morbidity or increasing the relative healthspan will require "delaying aging" or delaying the physiological change that results in disease and disability. While moving to life expectancies above age 95 and compressing morbidity substantially may require significant scientific breakthroughs; significant improvement in health and increases in life expectancy in the United States could be achieved with behavioral, life style, and policy changes that reduce socioeconomic disparities and allow us to reach the levels of health and life expectancy achieved in peer societies.


Introducing new data on gestation-specific infant mortality among babies born in 2005 in England and Wales.
Gestational age is highly correlated with birth outcomes including birthweight and infant mortality. Since gestational age is not recorded at the registration of live births in England and Wales, it has not been possible to produce routine statistics on gestation-specific infant mortality rates. A new system, introduced in 2002, for allocating NHS numbers at birth (NN4B) provided the opportunity to obtain gestational age information. NN4B records have been linked with birth registration data for all births occurring in 2005, and further linked with registration records for deaths in the first year of life. Thus, for the first time, we produce gestation-specific infant mortality rates for England and Wales as a whole, including in relation to birthweight, multiplicity, age of mother, marital status/registration type, and the National Statistics Socio-Economic Classification.


Thrombus Permeability Is Associated With Improved Functional Outcome and Recanalization in Patients With Ischemic Stroke.
Preclinical studies showed that thrombi can be permeable and may, therefore, allow for residual blood flow in occluded arteries of patients having acute ischemic stroke. This perviousness may increase tissue oxygenation, improve thrombus dissolution, and augment intra-arterial treatment success. We hypothesize that the combination of computed tomographic angiography and noncontrast computed tomography imaging allows measurement of contrast agent penetrating a permeable thrombus, and it is associated with improved outcome. Thrombus and contralateral artery attenuations in noncontrast computed tomography and computed tomographic angiography images were measured in 184 Multicenter Randomized Clinical trial of Endovascular treatment of acute ischemic stroke in the Netherlands (MR CLEAN) patients with thin-slice images. Two quantitative estimators of the thrombus permeability were introduced: computed tomographic angiography attenuation increase (Δ) and thrombus void fraction (ε). Patients were dichotomized as having a pervious or impervious thrombus and associated with outcome, recanalization, and final infarct volume. Patients with Δ≥10.9 HU (n=81 [44%]) and ε≥6.5% (n=77 [42%]) were classified as having a pervious thrombus. These patients were 3.2 (95% confidence interval, 1.7-6.4) times more likely to have a favorable outcome, and 2.5 (95% confidence interval, 1.3-4.8) times more likely to recanalyze, for Δ based classification, and similarly for ε. These odds ratios were independent from intravenous or intra-arterial treatment. Final infarct volume was negatively correlated with both perviousness estimates (correlation coefficient, -0.39 for Δ and -0.40 for ε). This study shows that simultaneous measurement of thrombus attenuation in noncontrast computed tomography and computed tomographic angiography allows for quantification of thrombus perviousness. Thrombus perviousness is strongly associated with improved functional outcome, smaller final infarct volume, and higher recanalization rate.


Recent advances in factors that alter lipid metabolism in chronic renal failure.
The following conclusions and speculations can be tentatively drawn from the changes in lipoprotein composition and metabolism: (1) The presence of apo B-48 in serum VLDL and the high serum apo A-IV concentrations indicate a greater than normal contribution of alimentary remnant particles to the hypertriglyceridemia of uremic patients, (2) The presence of apo E and C in triglyceride-enriched serum LDL, together with the triglyceride enrichment of all lipoproteins, probably stems from a deficiency of lipoprotein lipase (LPL) and hepatic lipase (HL) activity, (3) The decreased ratio of serum apo C-II/C-III in VLDL is at least in part responsible for the depressed activity of LPL, (4) The accumulation of lipoprotein particles with distorted apoprotein and lipid patterns (particularly beta-VLDL with enrichment in cholesterol) could be associated with an increased atherogenesis because a recent study has demonstrated a strong association between raised serum IDL and VLDL concentrations and the degree of coronary atherosclerosis, (5) The increased apo E content of VLDL and HDL in uremic patients could particularly point to a disturbed cholesterol metabolism because such lipoproteins could interact with LDL at apo B, E receptors, (6) The decrease in serum HDL-cholesterol has been shown to be strongly associated with atheromatous vascular disease, and this could also hold for uremic patients; however, it is probable that low serum HDL-cholesterol together with a diminished capacity to form cholesterol-rich, apo E containing HDL represents a decrease in the antiatherogenic defense of the organism rather than an increased atherogenic potential [21].


Interleukin-2 and regulatory T cells in graft-versus-host disease.
Dysfunction of regulatory T (Treg) cells has been detected in diverse inflammatory disorders, including chronic graft-versus-host disease (GVHD). Interleukin-2 is critical for Treg cell growth, survival, and activity. We hypothesized that low-dose interleukin-2 could preferentially enhance Treg cells in vivo and suppress clinical manifestations of chronic GVHD. In this observational cohort study, patients with chronic GVHD that was refractory to glucocorticoid therapy received daily low-dose subcutaneous interleukin-2 (0.3×10(6), 1×10(6), or 3×10(6) IU per square meter of body-surface area) for 8 weeks. The end points were safety and clinical and immunologic response. After a 4-week hiatus, patients with a response could receive interleukin-2 for an extended period. A total of 29 patients were enrolled. None had progression of chronic GVHD or relapse of a hematologic cancer. The maximum tolerated dose of interleukin-2 was 1×10(6) IU per square meter. The highest dose level induced unacceptable constitutional symptoms. Of the 23 patients who could be evaluated for response, 12 had major responses involving multiple sites. The numbers of CD4+ Treg cells were preferentially increased in all patients, with a peak median value, at 4 weeks, that was more than eight times the baseline value (P<0.001), without affecting CD4+ conventional T (Tcon) cells. The Treg:Tcon ratio increased to a median of more than five times the baseline value (P<0.001). The Treg cell count and Treg:Tcon ratio remained elevated at 8 weeks (P<0.001 for both comparisons with baseline values), then declined when the patients were not receiving interleukin-2. The increased numbers of Treg cells expressed the transcription factor forkhead box P3 (FOXP3) and could inhibit autologous Tcon cells. Immunologic and clinical responses were sustained in patients who received interleukin-2 for an extended period, permitting the glucocorticoid dose to be tapered by a mean of 60% (range, 25 to 100). Daily low-dose interleukin-2 was safely administered in patients with active chronic GVHD that was refractory to glucocorticoid therapy. Administration was associated with preferential, sustained Treg cell expansion in vivo and amelioration of the manifestations of chronic GVHD in a substantial proportion of patients. (Funded by a Dana-Farber Dunkin' Donuts Rising Star award and others; ClinicalTrials.gov number, NCT00529035.).


Plasmodium falciparum pfmdr1 amplification, mefloquine resistance, and parasite fitness.
Mefloquine is widely used in combination with artemisinin derivatives for the treatment of falciparum malaria. Mefloquine resistance in Plasmodium falciparum has been related to increased copy numbers of multidrug-resistant gene 1 (pfmdr1). We studied the ex vivo dynamics of pfmdr1 gene amplification in culture-adapted P. falciparum in relation to mefloquine resistance and parasite fitness. A Thai P. falciparum isolate (isolate TM036) was assessed by the use of multiple genetic markers as a single genotype. Resistance was selected by exposure to stepwise increasing concentrations of mefloquine up to 30 ng/ml in continuous culture. The pfmdr1 gene copy numbers increased as susceptibility to mefloquine declined (P = 0.03). No codon mutations at positions 86, 184, 1034, 1042, and 1246 in the pfmdr1 gene were detected. Two subclones of selected parasites (average copy numbers, 2.3 and 3.1, respectively) showed a fitness disadvantage when they were grown together with the original parasites containing a single pfmdr1 gene copy in the absence of mefloquine; the multiplication rates were 6.3% and 8.7% lower, respectively (P < 0.01). Modeling of the dynamics of the pfmdr1 copy numbers over time in relation to the relative fitness of the parasites suggested that net pfmdr1 gene amplification from one to two copies occurs once in every 10(8) parasites and that amplification from two to three copies occurs once in every 10(3) parasites. pfmdr1 gene amplification in P. falciparum is a frequent event and confers mefloquine resistance. Parasites with multiple copies of the pfmdr1 gene have decreased survival fitness in the absence of drug pressure.


Identification of serum biomarkers for canine B-cell lymphoma by use of surface-enhanced laser desorption-ionization time-of-flight mass spectrometry.
To identify biomarker proteins for B-cell lymphoma in canine serum by use of surface-enhanced laser desorption-ionization time-of-flight (SELDI-TOF) mass spectrometry and build classification trees with multiple biomarkers that have high sensitivity and specificity for that tumor type. Sera from 29 dogs with B-cell lymphoma and 87 control dogs (approx equal numbers of healthy dogs, dogs with malignant cancers other than B-cell lymphoma, and dogs with various nonneoplastic diseases or conditions). Serum samples were fractionated chromatographically and analyzed via SELDI-TOF mass spectrometry. Peak amplitudes of the spectra from the 2 sample groups were compared to identify potential biomarker peaks, and classification trees were built by use of computer software to detect patterns formed by multiple biomarkers among SELDI data sets. Several biomarker protein peaks in canine serum were identified, and a classification tree was built on the basis of 3 biomarker protein peaks. With 10-fold cross-validation of the sample set, the best individual serum biomarker peak had 75% sensitivity and 86% specificity and the classification tree had 97% sensitivity and 91% specificity for the classification of B-cell lymphoma. On the basis of biomarker proteins identified in canine serum, classification trees were constructed, which may be useful for the development of a diagnostic test for B-cell lymphoma in dogs. Further investigation is needed to determine whether these biomarkers are useful for screening susceptible dog populations or for monitoring disease status during treatment and remission of B-cell lymphoma in dogs.


Gene regulatory network reveals oxidative stress as the underlying molecular mechanism of type 2 diabetes and hypertension.
The prevalence of diabetes is increasing worldwide. It has been long known that increased rates of inflammatory diseases, such as obesity (OBS), hypertension (HT) and cardiovascular diseases (CVD) are highly associated with type 2 diabetes (T2D). T2D and/or OBS can develop independently, due to genetic, behavioral or lifestyle-related variables but both lead to oxidative stress generation. The underlying mechanisms by which theses complications arise and manifest together remain poorly understood. Protein-protein interactions regulate nearly every living process. Availability of high-throughput genomic data has enabled unprecedented views of gene and protein co-expression, co-regulations and interactions in cellular systems. The present work, applied a systems biology approach to develop gene interaction network models, comprised of high throughput genomic and PPI data for T2D. The genes differentially regulated through T2D were 'mined' and their 'wirings' were studied to get a more complete understanding of the overall gene network topology and their role in disease progression. By analyzing the genes related to T2D, HT and OBS, a highly regulated gene-disease integrated network model has been developed that provides useful functional linkages among groups of genes and thus addressing how different inflammatory diseases are connected and propagated at genetic level. Based on the investigations around the 'hubs' that provided more meaningful insights about the cross-talk within gene-disease networks in terms of disease phenotype association with oxidative stress and inflammation, a hypothetical co-regulation disease mechanism model been proposed. The results from this study revealed that the oxidative stress mediated regulation cascade is the common mechanistic link among the pathogenesis of T2D, HT and other inflammatory diseases such as OBS. The findings provide a novel comprehensive approach for understanding the pathogenesis of various co-associated chronic inflammatory diseases by combining the power of pathway analysis with gene regulatory network evaluation.


Aging and the role of the HPA axis and rhythm in sleep and memory-consolidation.
Changes in the hypothalamo-pituitary-adrenal (HPA) axis and its rhythm with aging have interesting implications for sleep. Herein, the authors review sleep and HPA changes associated with normal aging and point out the similarities in how they change over time. The authors also discuss the effects of sleep on declarative memory consolidation, in particular. This focused review suggests that some of the declarative memory dysfunction with normal aging, and possibly procedural memory dysfunction, may be partially reversible by instituting methods to augment slow-wave sleep (SWS). Also, agents that decrease nocturnal corticotropin-releasing hormone and the cortisol nadir and enhance SWS may offer potential ways to manipulate the HPA axis/rhythm and improve sleep and memory. In this regard, the authors propose that drugs that act directly on the HPA axis (e.g., mineralocorticoid agonists) may be potentially quite useful for improving both sleep and declarative memory consolidation during sleep.


Vascular inflammation in patients with impaired glucose tolerance and type 2 diabetes: analysis with 18F-fluorodeoxyglucose positron emission tomography.
Type 2 diabetes mellitus (T2DM) is associated with an increased risk of atherosclerotic cardiovascular disease. Vascular inflammation is a key factor in both the pathogenesis and outcome of atherosclerosis. (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) is a promising tool for indentifying and quantifying vascular inflammation within atherosclerotic plaques. This study was designed to examine the vascular inflammation measured using FDG-PET in patients with impaired glucose tolerance and T2DM, in comparison with age- and sex-matched control subjects with normal glucose tolerance. We investigated vascular inflammation using FDG-PET in 90 age- and sex-matched subjects with different glucose tolerance (30 normal glucose tolerance subjects, 30 impaired glucose tolerance subjects, and 30 T2DM subjects). Vascular 18F-FDG uptake was measured as both the mean and maximum blood-normalized standardized uptake value, known as the target-to-background ratio (TBR). Both mean and maximum TBR measurements were significantly different, based on glucose tolerance, although the carotid intima-media thickness measurements were not significantly different. The maximum TBR values in patients with impaired glucose tolerance and T2DM were significantly increased compared with the normal subjects. In addition, subjects with metabolic syndrome had increased maximum TBR values compared with those without metabolic syndrome. Age-, sex-, and body mass index-adjusted maximum TBR levels were positively correlated with triglyceride, hemoglobin A1c, insulin resistance, high-sensitivity C-reactive protein, and Framingham risk score and were negatively correlated with high-density lipoprotein cholesterol and adiponectin levels. The results of the present study suggest that impaired glucose tolerance and T2DM are associated with vascular inflammation in carotid atherosclerosis detected by FDG-PET.


Definitions and potential health benefits of the Mediterranean diet: views from experts around the world.
The Mediterranean diet has been linked to a number of health benefits, including reduced mortality risk and lower incidence of cardiovascular disease. Definitions of the Mediterranean diet vary across some settings, and scores are increasingly being employed to define Mediterranean diet adherence in epidemiological studies. Some components of the Mediterranean diet overlap with other healthy dietary patterns, whereas other aspects are unique to the Mediterranean diet. In this forum article, we asked clinicians and researchers with an interest in the effect of diet on health to describe what constitutes a Mediterranean diet in different geographical settings, and how we can study the health benefits of this dietary pattern.


A marked reduction in priming of cytotoxic CD8+ T cells mediated by stress-induced glucocorticoids involves multiple deficiencies in cross-presentation by dendritic cells.
Protracted psychological stress elevates circulating glucocorticoids, which can suppress CD8(+) T cell-mediated immunity, but the mechanisms are incompletely understood. Dendritic cells (DCs), required for initiating CTL responses, are vulnerable to stress/corticosterone, which can contribute to diminished CTL responses. Cross-priming of CD8(+) T cells by DCs is required for initiating CTL responses against many intracellular pathogens that do not infect DCs. We examined the effects of stress/corticosterone on MHC class I (MHC I) cross-presentation and priming and show that stress/corticosterone-exposed DCs have a reduced ability to cross-present OVA and activate MHC I-OVA(257-264)-specific T cells. Using a murine model of psychological stress and OVA-loaded β(2)-microglobulin knockout "donor" cells that cannot present Ag, DCs from stressed mice induced markedly less Ag-specific CTL proliferation in a glucocorticoid receptor-dependent manner, and endogenous in vivo T cell cytolytic activity generated by cross-presented Ag was greatly diminished. These deficits in cross-presentation/priming were not due to altered Ag donation, Ag uptake (phagocytosis, receptor-mediated endocytosis, or fluid-phase uptake), or costimulatory molecule expression by DCs. However, proteasome activity in corticosterone-treated DCs or splenic DCs from stressed mice was partially suppressed, which limits formation of antigenic peptide-MHC I complexes. In addition, the lymphoid tissue-resident CD11b(-)CD24(+)CD8α(+) DC subset, which carries out cross-presentation/priming, was preferentially depleted in stressed mice. At the same time, CD11b(-)CD24(+)CD8α(-) DC precursors were increased, suggesting a block in development of CD8α(+) DCs. Therefore, glucocorticoid-induced changes in both the cellular composition of the immune system and intracellular protein degradation contribute to impaired CTL priming in stressed mice.


Cognitive effects of exogenous melatonin administration in elderly persons: a pilot study.
Given that circadian rhythm disruption is associated with impairments in cognitive performance similar to those found in age-related cognitive decline, the authors investigated whether exogenous melatonin administration would improve cognitive functioning in healthy elderly subjects. This double-blind, placebo-controlled pilot study assigned 26 healthy elderly subjects to receive either melatonin 1 mg or placebo nightly for 4 weeks. Participants completed a sleep questionnaire and a battery of cognitive tests at baseline and at 4 weeks. Melatonin administration improved reported morning "restedness" and sleep latency after nocturnal awakening, and also improved scores on the California Verbal Learning Test-interference subtest. Melatonin administration at a dose of 1 mg nightly may be effective in improving certain aspects of cognitive functioning and subjective reports of sleep quality in elderly subjects. It may prove to be a useful therapeutic agent in the treatment of age-related cognitive decline.


Skeletal Muscle and Metabolic Health: How Do We Increase Muscle Mass and Function in People with Type 2 Diabetes?
Whilst skeletal muscles' primary role is allowing movement, it has important metabolic roles, including in glycemic control. Indeed, evidence indicates that low muscle mass and function are associated with an increased risk of type 2 diabetes, highlighting its importance in the development of metabolic disease. In this mini-review, we detail the evidence highlighting the importance of muscle in type 2 diabetes and the efficacy of resistance exercise in improving glycemic control alongside our approach to increase uptake of such exercise in people with type 2 diabetes. This summary is based in the authors' knowledge of the filed supplemented by a Pubmed search using the terms "muscle," "glycemic control," "HbA1c," "type 2 diabetes," and "resistance exercise." The main strategy to increases muscle mass is to perform resistance exercise and, although the quality of evidence is low, such exercise appears effective in reducing Glycated Haemoglobin (HbA1c) in people with type 2 diabetes. However, to increase participation we need to improve our understanding of barriers and facilitators to such exercise. Current data indicate that barriers are similar to those reported for aerobic exercise, with additional resistance exercise specific barriers of looking to muscular, increase risk of cardiovascular event, having access to specialized equipment and knowledge of how to use it. The development of simple resistance exercises that can be performed anywhere, that use little or no equipment and are effective in reducing HbA1c will be, in our opinion, key to increasing the number of people with type 2 diabetes performing resistance exercise.


Cage-templated synthesis of highly stable palladium nanoparticles and their catalytic activities in Suzuki-Miyaura coupling.
We report the controlled synthesis of small palladium nanoparticles (PdNPs) with narrow particle size distribution (1.8 ± 0.2 nm) using an organic molecular cage as a template. The well-defined cage structure and thioether anchoring groups inside the cavity are critical for the formation of narrowly distributed PdNPs, offering a confined organic molecular environment and guiding PdNP nucleation and growth. The resulting encapsulated PdNPs are resistant to agglomeration and stable in solution exposed to air at room temperature. When provided with a protective cage shell with minimum surface coverage, such PdNPs are capable of catalyzing organic reactions, showing high catalytic activity in Suzuki-Miyaura coupling reactions.


Evolutionary trade-offs between male secondary sexual traits revealed by a phylogeny of the hyperdiverse tribe Eumaeini (Lepidoptera: Lycaenidae).
Male butterflies in the hyperdiverse tribe Eumaeini possess an unusually complex and diverse repertoire of secondary sexual characteristics involved in pheromone production and dissemination. Maintaining multiple sexually selected traits is likely to be metabolically costly, potentially resulting in trade-offs in the evolution of male signals. However, a phylogenetic framework to test hypotheses regarding the evolution and maintenance of male sexual traits in Eumaeini has been lacking. Here, we infer a comprehensive, time-calibrated phylogeny from 379 loci for 187 species representing 91% of the 87 described genera. Eumaeini is a monophyletic group that originated in the late Oligocene and underwent rapid radiation in the Neotropics. We examined specimens of 818 of the 1096 described species (75%) and found that secondary sexual traits are present in males of 91% of the surveyed species. Scent pads and scent patches on the wings and brush organs associated with the genitalia were probably present in the common ancestor of Eumaeini and are widespread throughout the tribe. Brush organs and scent pads are negatively correlated across the phylogeny, exhibiting a trade-off in which lineages with brush organs are unlikely to regain scent pads and vice versa. In contrast, scent patches seem to facilitate the evolution of scent pads, although they are readily lost once scent pads have evolved. Our results illustrate the complex interplay between natural and sexual selection in the origin and maintenance of multiple male secondary sexual characteristics and highlight the potential role of sexual selection spurring diversification in this lineage.


Heart rate (variability) and the association between relational peer victimization and internalizing symptoms in elementary school children.
Relational victimization typically emerges first during the elementary school period, and has been associated with increased levels of internalizing symptoms in children. Individual differences in autonomic nervous system functioning have been suggested as a potential factor linking social stressors and internalizing symptoms. The aim of this study was therefore to examine whether heart rate and heart rate variability mediated the association between relational victimization and internalizing symptoms in 373 mainstream elementary school children. Children were assessed in 2015 (T0; Grades 3-5, M age = 9.78 years, 51% boys) and reassessed in 2016 (T1). Heart rate and heart rate variability were assessed during a regular school day at T1. A multi-informant (teacher and peer report) cross-time measure of relational victimization, and a multi-informant (self- and teacher report) measure of internalizing problems at T1 was used. Results showed that heart rate variability, but not heart rate, mediated the association between relational victimization and internalizing symptoms. This study provides tentative support that in children from a general population sample, a psychobiological factor may mediate the association of relational victimization with internalizing symptoms.


Perinatal Exposure to Glyphosate and a Glyphosate-Based Herbicide Affect Spermatogenesis in Mice.
Glyphosate is the most widely used herbicide in the world. Several studies have investigated the effects of glyphosate and glyphosate-based herbicides (GBHs) on male reproduction, but there is still little and conflicting evidence for its toxicity. In this study, we analyzed the effects of glyphosate, alone or in formula, on the male reproductive system. Pregnant mice were treated from E10.5 to 20 days postpartum by adding glyphosate or a GBH (Roundup 3 Plus) to their drinking water at 0.5 (the acceptable daily intake, ADI dose), 5 and 50 mg/kg/day. Male offspring derived from treated mice were sacrificed at 5, 20, and 35 days old (d.o.) and 8 months old (m.o.) for analysis. Our result showed that exposure to glyphosate, but not GBH, affects testis morphology in 20 d.o. and decrease serum testosterone concentrations in 35 d.o. males. We identified that the spermatozoa number decreased by 89% and 84% in 0.5 and 5 mg/kg/day of GBH and glyphosate groups, respectively. Moreover, the undifferentiated spermatogonia numbers were decreased by 60% in 5 mg/kg/day glyphosate group, which could be due to the alterations in the expression of genes involved in germ cell differentiation such as Sall4 and Nano3 and apoptosis as Bax and Bcl2. In 8 m.o. animals, a decreased testosterone level was observed in GBH groups. Our data demonstrate that glyphosate and GBHs could cause endocrine-disrupting effects on male reproduction at low doses. As glyphosate has effects at the ADI level, our data suggest that the current ADI for glyphosate could be overestimated.


Evidence for an Intermediate in the Methylation of CB11 H12 - with Methyl Triflate: Comparison of Electrophilic Substitution in Cage Boranes and in Arenes.
The trideuteriomethylation of BH vertices in CB11 H12 - and its derivatives with CD3 OTf (OTf=triflate, trifluoromethanesulfonate) yields a mixture of BCD3 and BCHD2 substitution products, thus demonstrating the intermediacy of a species with a long enough lifetime for hydrogen scrambling between the boron vertex and the methyl substituent. No such scrambling is observed if CD3 OTf is used to methylate toluene. According to density functional theory calculations, the intermediate in BH vertex methylation is a three-center bonded σ adduct of a methyl cation to the BH bond and the proton scrambling occurs via a transition structure containing a distorted square-pyramidal methane attached axially to a "naked" boron vertex. The subsequent proton or deuteron loss is presently not understood in detail. A general comparison of electrophilic substitution on closo-boranes and arenes is provided and similarities as well as differences are discussed. A recalculation of the optimized geometry of the CB11 Me12 . radical produced a second Jahn-Teller distorted minimum and resulted in a somewhat improved agreement between calculated and measured proton hyperfine coupling constants.


High-resolution characterization of short-term temporal variability in the taxonomic and resistome composition of wastewater influent.
Wastewater-based epidemiology (WBE) for population-level surveillance of antimicrobial resistance (AMR) is gaining significant traction, but the impact of wastewater sampling methods on results is unclear. In this study, we characterized taxonomic and resistome differences between single-timepoint-grab and 24 h composites of wastewater influent from a large UK-based wastewater treatment work [WWTW (population equivalent: 223 435)]. We autosampled hourly influent grab samples (n=72) over three consecutive weekdays, and prepared additional 24 h composites (n=3) from respective grabs. For taxonomic profiling, metagenomic DNA was extracted from all samples and 16S rRNA gene sequencing was performed. One composite and six grabs from day 1 underwent metagenomic sequencing for metagenomic dissimilarity estimation and resistome profiling. Taxonomic abundances of phyla varied significantly across hourly grab samples but followed a repeating diurnal pattern for all 3 days. Hierarchical clustering grouped grab samples into four time periods dissimilar in both 16S rRNA gene-based profiles and metagenomic distances. 24H-composites resembled mean daily phyla abundances and showed low variability of taxonomic profiles. Of the 122 AMR gene families (AGFs) identified across all day 1 samples, single grab samples identified a median of six (IQR: 5-8) AGFs not seen in the composite. However, 36/36 of these hits were at lateral coverage <0.5 (median: 0.19; interquartile range: 0.16-0.22) and potential false positives. Conversely, the 24H-composite identified three AGFs not seen in any grab with higher lateral coverage (0.82; 0.55-0.84). Additionally, several clinically significant human AGFs (bla VIM, bla IMP, bla KPC) were intermittently or completely missed by grab sampling but captured by the 24 h composite. Wastewater influent undergoes significant taxonomic and resistome changes on short timescales potentially affecting interpretation of results based on sampling strategy. Grab samples are more convenient and potentially capture low-prevalence/transient targets but are less comprehensive and temporally variable. Therefore, we recommend 24H-composite sampling where feasible. Further validation and optimization of WBE methods is vital for its development into a robust AMR surveillance approach.


Wrist Arthroscopy: Can We Gain Proficiency Through Knee Arthroscopy Simulation?
Wrist arthroscopy is a challenging discipline with limited training exposure during residency. The purpose of this study was to evaluate the effectiveness of virtual knee arthroscopy simulation training for gaining proficiency in wrist arthroscopy. Participants were recorded performing a cadaveric wrist arthroscopy simulation. The residents then practiced knee arthroscopy on a virtual reality simulator and repeated the wrist arthroscopy simulation. All videos were blinded prior to assessment. Proficiency was graded using the Arthroscopic Surgery Skill Evaluation Tool global rating scale. In addition, participants were asked to complete a survey assessing the value of the virtual reality knee arthroscopy simulator for wrist arthroscopy. Orthopaedic Surgery Residency Program, Carolinas Medical Center, a large, public, nonprofit hospital located in Charlotte, North Carolina. Orthopaedic residents at our center were asked to participate in the simulation training. Participation was voluntary and nonincentivized. All orthopaedic residents at our institution (N = 27) agreed to participate. In total, there were 10 Intern (PGY-0 and PGY-1), 10 Junior (PGY-2 and PGY-3), and 7 Senior (PGY-4 and PGY-5) residents. In addition, a fellowship-trained hand surgeon was recruited to participate in the study, performing the wrist arthoscopy simulation. Two additional fellowship-trained hand surgeons, for a total of 3, assessed the blinded videos. There was a trend toward better wrist Arthroscopic Surgery Skill Evaluation Tool scores by training level, although the difference was not statistically significant. Interns improved by an average of 1.8 points between baseline and postknee simulation tests. Junior and senior residents decreased by 1.6 and 5.0 points, respectively. Knee arthroscopy simulation training did not objectively improve wrist arthroscopy proficiency among residents. A wrist-specific arthroscopy simulation program is needed if measurable competence through simulation is desired.


Red meat intake, insulin resistance, and markers of endothelial function among Iranian women.
Few data, with conflicting findings, are available linking red meat consumption to indicators of insulin resistance and endothelial dysfunction. This study aimed to investigate the association of red meat consumption with insulin resistance and endothelial dysfunction among a sample of female nurses in Isfahan, Iran. This cross-sectional study was carried out among 420 female nurses who were selected by a multistage cluster random sampling method. Usual dietary intakes were assessed using a validated food frequency questionnaire. Red meat intake was calculated by summing up the consumption of all kinds of red meat in foods and processed meat in sausages and fast foods. To measure serum concentrations of adhesion molecules and glycemic indexes, a fasting blood sample was taken. After adjustment for potential confounders, high red meat intake was significantly associated with higher fasting plasma glucose, homeostasis model assessment of insulin resistance, and lower quantitative insulin sensitivity check index. Although high red meat intake was significantly associated with higher serum insulin levels and lower homeostasis model assessment of beta-cell function in the crude model, after controlling for BMI, the association was no longer significant. Red meat consumption was associated with high concentrations of E-selectin, soluble vascular cell adhesion molecule-1 (sVCAM-1), and soluble intercellular adhesion molecule-1 (sICAM-1) after adjustment for different potential confounders. We found that increased red meat intake was associated with high concentrations of plasma endothelial dysfunction biomarkers and abnormal glucose homeostasis among Iranian women. Prospective studies are required to confirm these findings.


Tn5 transposase mutants that alter DNA binding specificity.
Tn5 transposase (Tnp) binds to Tn5 and IS50 end inverted repeats, the outside end (OE) and the inside end (IE), to initiate transposition. We report the isolation of four Tnp mutants (YH41, TP47, EK54 and EV54) that increase the OE-mediated transposition frequency and enhance the binding affinity of Tnp for OE DNA. In addition, two of the Tnp mutants (TP47 and EK54) appear to be change-of-specificity mutants, since they alter the recognition of OE versus IE relative to the wild-type Tnp. EK54 enhances OE recognition but decreases IE recognition. TP47 enhances both OE and IE recognition but with a much greater enhancement for IE than for OE. This change-of-specificity effect of TP47 is observed only when TP47 Tnp is synthesized in cis to the DNA that contains the ends. We propose that Lys54 makes a favorable interaction with an OE-specific nucleotide pair(s), while Pro47 may cause a more favorable interaction with an IE-specific nucleotide pair(s) than it does with the corresponding OE-specific nucleotide pair(s). A model to explain the preference of TP47 Tnp for the IE in cis but not in trans is proposed.


The metabolism and elimination of pyrilamine maleate in the rat.
The metabolism and elimination of pyrilamine (2-[(2-dimethylamino-ethyl)(p-methoxybenzyl)amino]pyridine) were characterized after the iv administration of 7.0 mg/kg or 0.7 mg/kg pyrilamine maleate plus [14C]pyrilamine maleate to adult male Fischer-344 rats. Approximately 29% and 38% of the administered dose was excreted in the urine in the first 24 hr in the high and low dose groups, respectively, as determined by liquid scintillation spectrometry. Fecal excretion accounted for 27% and 30% of the administered dose in the first 24 hr in the high and low dose groups, respectively, as confirmed via combustion analysis. The 24-hr urinary metabolic products consisted of one major and four minor radiolabeled compounds. The major metabolite was isolated by reversed-phase high performance liquid chromatography and identified as the O-glucuronic acid conjugate of O-demethyl pyrilamine. This was accomplished by comparison of the chromatographic characteristics of this metabolite's aglycon with that of an authentic standard of O-demethyl pyrilamine and fast atom bombardment mass spectrometry of the unhydrolyzed conjugate. Pyrilamine and its N-oxide and O-demethyl derivatives were also identified after isolation by reversed-phase high performance liquid chromatography and comparison of their mass spectral and/or chromatographic properties with those of authentic compounds. The plasma metabolic profile was essentially the same as the urinary profile except for the absence of O-demethyl pyrilamine. The plasma elimination of pyrilamine fit a one-compartment open model and was first order. The terminal plasma elimination half-life of pyrilamine did not increase with increasing doses (2.3 hr, 0.7 mg/kg; 1.5 hr, 7.0 mg/kg) and thus pyrilamine does not exhibit dose-dependent elimination.(ABSTRACT TRUNCATED AT 250 WORDS)


Spatial analysis of germ stem cell development in Oct-4/EGFP transgenic mice.
Questions persist regarding male germ stem cells and how they mature during the prespermatogenic period of testicular development. We successfully labeled the prespermatogonia with green fluorescence protein (GFP) by using Oct-4 enhancer/promoter. This study shows that GFP was specifically expressed in prespermatogonia, spermatogonia and spermatids that faithfully reproduce the endogenous expression of Oct-4. Histochemical analysis revealed that most of the TRA98-positive gonocytes are also positive for GFP. However, the frequency of GFP expressing cells out of TRA98 expressing cells decreased together with the maturation of gonocytes in the first week after birth. To compare the stem cell activity between GFP-positive and -negative populations, we performed a transplantation of sorted cells into testes from an individual population. Colonization efficiency of germ cells from a GFP-positive population resulted in a 30-fold increase in colonization compared with a GFP-negative population. Since the expression of Oct-4 in prespermtogonia correlates well with the stem activity, Oct-4 might be a crucial molecule in the stem cell property of spermatogonia but not in cell survival.


The rate of purine synthesis de nova in blood mononuclear cells in vitro from patients with familial hyperuricaemic nephropathy.
We have measured the rate of purine synthesis de novo in blood mononuclear cells in vitro and the activities of the purine salvage enzymes [hypoxanthine phosphoribosyltransferase (HPRT; EC 126.96.36.199), adenine phosphoribosyltransferase (APRT; EC 220.127.116.11)] and ribosephosphate pyrophosphokinase (PP-ribose-P synthetase; EC 188.8.131.52)] and the concentration of phosphoribosylpyrophosphate (PP-ribose-P) in the erythrocytes of affected family members. These subjects belong to families where hyperuricaemia and renal failure occur together early in life, and the genetic transmission follows an autosomal dominant mode of inheritance. We term this syndrome, familial hyperuricaemic nephropathy. No significant differences were detected in either the rates of purine synthesis de novo in vitro between the index patients and the control subjects with respect to the enzyme activities or the PP-ribose-P concentrations. Two groups of controls were used, healthy individuals and patients with a comparable degree of renal failure due to non-immune complex renal disease. Mononuclear cells from patients with Lesch-Nyhan syndrome (congenital HPRT deficiency) showed the expected acceleration of purine synthesis de novo in vitro. The accelerated purine synthesis de novo in vitro associated with phytohaemagglutinin-induced lymphocyte transformation was detectable by the method used. We conclude that familial hyperuricaemic nephropathy is not due to a metabolic lesion which causes accelerated purine synthesis de novo. This suggests that the primary abnormality may be a failure of the renal tubular net excretion of urate.


A piRNA pathway primed by individual transposons is linked to de novo DNA methylation in mice.
piRNAs and Piwi proteins have been implicated in transposon control and are linked to transposon methylation in mammals. Here we examined the construction of the piRNA system in the restricted developmental window in which methylation patterns are set during mammalian embryogenesis. We find robust expression of two Piwi family proteins, MIWI2 and MILI. Their associated piRNA profiles reveal differences from Drosophila wherein large piRNA clusters act as master regulators of silencing. Instead, in mammals, dispersed transposon copies initiate the pathway, producing primary piRNAs, which predominantly join MILI in the cytoplasm. MIWI2, whose nuclear localization and association with piRNAs depend upon MILI, is enriched for secondary piRNAs antisense to the elements that it controls. The Piwi pathway lies upstream of known mediators of DNA methylation, since piRNAs are still produced in dnmt3L mutants, which fail to methylate transposons. This implicates piRNAs as specificity determinants of DNA methylation in germ cells.


Osseous lesions in the metacarpo(tarso)phalangeal joint diagnosed using low-field magnetic resonance imaging in standing horses.
We report the use of low-field standing magnetic resonance imaging in the standing horse for the diagnosis of osseous lesions in the metacarpophalangeal (MCP) or metatarsophalangeal (MTP) joint that were not apparent using standard radiography. Thirteen horses were studied and all had thickening of the subchondral bone plate and abnormal signal intensity in the adjacent spongiosa in either the condyles of metacarpal/metatarsal III or the proximal phalanx or both. Abnormalities were characterized by diffuse decreased signal intensity on T1-weighting adjacent to the subchondral bone and within the spongiosa in at least two imaging planes; in the absence of increases in signal intensity in fat-suppressed images, this change was interpreted as bone sclerosis. Nine horses also had a diffuse decreased signal intensity on T2*-weighting in the same areas and five had a diffuse increase in signal intensity in fat-suppressed images in conjunction with a decrease in signal intensity on T1- and T2*-weighted images; the increase in signal intensity in fat-suppressed images was interpreted as fluid accumulation. Five horses had a focal area of change in signal intensity within the subchondral bone with apparent loss of definition between the subchondral bone and the articular cartilage. Eleven horses were available for follow up, of which eight were sound and three remained lame. We conclude that lameness originating from the MCP or MTP joint may be associated with osseous damage in horses of any signalment in the absence of radiographic changes.


Site-specific induction of intestinal hypoxia-inducible factor-1α after hemorrhagic shock.
The intestine is a major target organ in hemorrhagic shock (HS)-induced tissue injury. Hypoxia-inducible factor (HIF)-1α is the primary transcription factor responsible for regulating cellular response to changes in oxygen tension. Since HS is an acute hypoxic insult, the present study examined changes in the gene expression of HIF-1α in various regions of the intestine, as well as the distribution of HIF-1α protein in the intestinal cells of a rat model of HS. Levels of HIF-1α mRNA were marginally detectable in the intestine of sham-operated control animals, but obviously induced following HS. Duodenal, jejunal and colonic levels of HIF-1α mRNA robustly increased and reached a maximum during the ischemic phase of HS, followed by a rapid decrease almost to control levels during the early phase of resuscitation. The induction of HIF-1α mRNA was maximal in the duodenum. In contrast to the duodenum, jejunum and colon, in the ileum the HIF-1α mRNA level did not increase after HS. Consistent with enhanced HIF-1α gene expression, HIF-1α protein was expressed in the mucosal cells of the duodenum, jejunum and colon, but not in the ileum following HS. These findings indicate that intestinal HIF-1α expression was up-regulated at both the transcriptional and protein level in a site-specific manner in this rat model of HS. Differential regulation of HIF-1α expression along the longitudinal axes of the intestine might be a determinant of the adaptive response to HS-induced intestinal damage.


Radiographers' Ability to Detect Low-Contrast Detail in Digital Radiography Systems.
To evaluate radiographers' ability to detect low-contrast detail using various digital planar radiographic systems. A low-contrast detail phantom was placed between two 5-cm thick Perspex sheets (Lucite International). Images were obtained using different kilovoltage peak and milliamperage second (mAs) settings with computed radiography (CR), indirect conversion digital radiography (IDR), and direct conversion digital radiography (DR) systems. Six groups of 6 radiographers were asked to score 39 images; each group scored 2 images from each system for a total of 6 images. The seventh group scored only one image from each system for a total of 3 images. The radiographers' results were compared with the results of analyzer software. The inverse image quality factor was used to measure low-contrast detail detectability performance. Radiographers performed significantly worse than the computerized software in determining low-contrast detail in planar radiographic images (P < .01). However, a positive correlation (R = 0.558) existed between the 2 sets of scores in terms of low-contrast detail detectability performance. On average, radiographers were able to detect increased image quality resulting from increased mAs. Radiographers reached results similar to the software regarding whether IDR and DR have better detectability performances than CR. Differences found among individual radiographers were not as significant with DR. When radiographers' performance in detecting low-contrast detail was evaluated and compared with that of the software, radiographers exhibited poorer performances. Because radiographers are responsible for image quality optimization, additional training might improve their ability to detect low-contrast detail in DR systems.