The interaction of PP1 with BRCA1 and analysis of their expression in breast tumors.
The breast cancer susceptibility gene, BRCA1, is implicated in multiple cellular processes including DNA repair, the transactivation of genes, and the ubiquitination of proteins; however its precise functions remain to be fully understood. Identification and characterization of BRCA1 protein interactions may help to further elucidate the function and regulation of BRCA1. Additionally, detection of changes in the expression levels of BRCA1 and its interacting proteins in primary human breast tumors may further illuminate their role in the development of breast cancer. We performed a yeast two-hybrid study to identify proteins that interact with exon11 of BRCA1 and identified Protein Phosphatase 1beta (PP1beta), an isoform of the serine threonine phosphatase, PP1. GST-pull down and co-immunoprecipitation assays were performed to further characterize this interaction. Additionally, Real-Time PCR was utilized to determine the expression of BRCA1, PP1alpha, beta and gamma in primary human breast tumors and normal breast tissue to identify alterations in the expression of these genes in breast cancer. PP1 and BRCA1 co-immunoprecipitate and the region within BRCA1 as well as the specific PP1 interacting domain mediating this interaction were identified. Following mRNA expression analysis, we identified low levels of BRCA1 and variable levels of PP1alpha and beta in primary sporadic human breast tumors. Furthermore, BRCA1, PP1beta and PP1gamma were significantly higher in normal tissue specimens (BRCA1 p = 0.01, PP1beta: p = 0.03, PP1gamma, p = 1.9 x 10(-6)) compared to sporadic breast tumor samples. Interestingly, we also identified that ER negative tumors are associated with low levels of PP1alpha expression. The identification and characterization of the interaction of BRCA1 with PP1 and detection of changes in the expression of PP1 and genes encoding other BRCA1 associated proteins identifies important genetic pathways that may be significant to breast tumorigenesis. Alterations in the expression of genes, particularly phosphatases that operate in association with BRCA1, could negatively affect the function of BRCA1 or BRCA1 associated proteins, contributing to the development of breast cancer.


Acute hepatitis C: clinical aspects, diagnosis, and outcome of acute HCV infection.
Acute hepatitis C virus (HCV) infection is often a clinically silent infection, and is therefore rarely detected. A high index of clinical suspicion in addition to careful serological and virological assessment is required to identify the disease, and to determine the eventual clinical outcome after primary infection; the minority of acutely infected individuals spontaneously control viremia in long term whilst the majority become persistently infected. Here, we describe the clinical presentation of acute HCV infection and the patterns of viremia and liver alanine transaminase levels (ALT) observed. We discuss the serological and virological assessment and potential pitfalls in accurately diagnosing acute HCV. Good prospective studies that identify host and virological factors that determine clinical symptoms and disease outcome are difficult to perform due to the asymptomatic nature of infection, but some progress has been made in this field. Host factors including gender, age at time of infection, prior resolution of infection, symptomatic infection and host immune responses, and viral factors such as the nature of the infecting quasispecies and more speculatively viral genotype, are some features that have been correlated with disease outcome. In spite of this, on an individual patient level, it is currently not possible to predict those that will resolve infection. Identifying, in detail therefore, those factors that are responsible for viral control remains an important research goal not only to aid clinical management but also to develop effective treatment and vaccination strategies.


Green tea extract ingestion, fat oxidation, and glucose tolerance in healthy humans.
Green tea consumption is reportedly associated with various health-promoting properties. For example, it has been shown to promote fat oxidation in humans at rest and to prevent obesity and improve insulin sensitivity in mice. We investigated the effects of acute ingestion of green tea extract (GTE) on glucose tolerance and fat oxidation during moderate-intensity exercise in humans. Two studies were performed, both with a counter-balanced crossover design. In study A, 12 healthy men performed a 30-min cycling exercise at 60% of maximal oxygen consumption (VO2max) before and after supplementation. In study B, 11 healthy men took an oral-glucose-tolerance test before and after supplementation. In the 24-h period before the experimental trials, participants ingested 3 capsules containing either GTE (total: 890 +/- 13 mg polyphenols and 366 +/- 5 mg EGCG) or a corn-flour placebo (total: 1729 +/- 22 mg). Average fat oxidation rates were 17% higher after ingestion of GTE than after ingestion of placebo (0.41 +/- 0.03 and 0.35 +/- 0.03 g/min, respectively; P < 0.05). Moreover, the contribution of fat oxidation to total energy expenditure was also significantly higher, by a similar percentage, after GTE supplementation. The insulin area under the curve decreased in both the GTE and placebo trials (3612 +/- 301 and 4280 +/- 309 microIU/dL . 120 min, respectively; P < 0.01), and there was a concomitant increase of 13% in insulin sensitivity. Acute GTE ingestion can increase fat oxidation during moderate-intensity exercise and can improve insulin sensitivity and glucose tolerance in healthy young men.


Developmental abnormalities in cortical GABAergic system in mice lacking mGlu3 metabotropic glutamate receptors.
Polymorphic variants of the gene encoding for metabotropic glutamate receptor 3 (mGlu3) are linked to schizophrenia. Because abnormalities of cortical GABAergic interneurons lie at the core of the pathophysiology of schizophrenia, we examined whether mGlu3 receptors influence the developmental trajectory of cortical GABAergic transmission in the postnatal life. mGlu3-/- mice showed robust changes in the expression of interneuron-related genes in the prefrontal cortex (PFC), including large reductions in the expression of parvalbumin (PV) and the GluN1 subunit of NMDA receptors. The number of cortical cells enwrapped by perineuronal nets was increased in mGlu3-/- mice, suggesting that mGlu3 receptors shape the temporal window of plasticity of PV+ interneurons. Electrophysiological measurements of GABAA receptor-mediated responses revealed a more depolarized reversal potential of GABA currents in the somata of PFC pyramidal neurons in mGlu3-/- mice at postnatal d 9 associated with a reduced expression of the K+/Cl- symporter. Finally, adult mGlu3-/- mice showed lower power in electroencephalographic rhythms at 1-45 Hz in quiet wakefulness as compared with their wild-type counterparts. These findings suggest that mGlu3 receptors have a strong impact on the development of cortical GABAergic transmission and cortical neural synchronization mechanisms corroborating the concept that genetic variants of mGlu3 receptors may predispose to psychiatric disorders.-Imbriglio, T., Verhaeghe, R., Martinello, K., Pascarelli, M. T., Chece, G., Bucci, D., Notartomaso, S., Quattromani, M., Mascio, G., Scalabrì, F., Simeone, A., Maccari, S., Del Percio, C., Wieloch, T., Fucile, S., Babiloni, C., Battaglia, G., Limatola, C., Nicoletti, F., Cannella, M. Developmental abnormalities in cortical GABAergic system in mice lacking mGlu3 metabotropic glutamate receptors.


Increased Ca2+ signaling after alpha-adrenoceptor activation in vascular hypertrophy.
In an effort to explain the increased sensitivity to agonists of hypertrophic vascular muscle, intracellular Ca2+ concentration ([Ca2+]i)-signaling mechanisms were studied in normal and hypertrophic rat aortas from normotensive and coarctation-hypertensive rats. Based on both fura 2 fluorescence and aequorin luminescence measurements, qualitatively different patterns of Ca2+ mobilization occur in normal and hypertrophic rat aortic muscle. Normal rat aortic muscle contracts to phenylephrine with little or no increase in [Ca2+]i, whereas the angiotensin II-induced contraction is accompanied by a marked [Ca2+]i transient. In contrast, hypertrophic rat aortic muscle shows a dramatic increase in Ca2+ signaling after phenylephrine stimulation. Moreover, both the amplitude of the angiotensin-induced [Ca2+]i transient and the contractile sensitivity to this agonist are decreased in the hypertrophic muscle. Our results strongly suggest that the amplitude of the [Ca2+]i transient after agonist stimulation determines the contractile sensitivity and that there is an altered coupling of the alpha-adrenoceptor in the hypertrophic vascular muscle.


The cost of privatization: extra payments to Medicare Advantage plans--updated and revised.
The Medicare Modernization Act of 2003 sharply increased payments to private Medicare Advantage plans. As a result, every plan in every county in the nation was paid more in 2005 than its enrollees would have been expected to cost if they had been enrolled in traditional fee-for-service Medicare. The authors calculate that payments to Medicare Advantage plans averaged 12.4 percent more than costs in traditional Medicare during 2005: a total of more than $5.2 billion, or $922 for each of the 5.6 million Medicare enrollees in managed care. This issue brief updates an earlier analysis of Medicare Advantage payments in 2005 previously published by The Commonwealth Fund; the updated estimates in this report are based on final 2005 enrollment figures that were not available at the time the previous estimates were developed, and they include the effect of policy decisions that were not reflected in the previous estimates.


Age and motivation predict gaze behavior for facial expressions.
This study investigated age-related differences between younger (M = 25.52 years) and older (M = 70.51 years) adults in avoidance motivation and the influence of avoidance motivation on gaze preferences for happy, neutral, and angry faces. In line with the hypothesis of reduced negativity effect later in life, older adults avoided angry faces and (to a lesser degree) preferred happy faces more than younger adults did. This effect cannot be explained by age-related changes in dispositional motivation. Irrespective of age, avoidance motivation predicted gaze behavior towards emotional faces. The study demonstrates the importance of interindividual differences beyond young adulthood.


Single cell protein as an occupational hazard.
Single cell protein (SCP) intended for animal feed purposes was produced in a pilot plant. The SCP consisted of Methylomonas methanolica, a pseudomonas species which is an obligate methanol user. The SCP was cultured in fermenters and later dewatered and dried in a spray-drier. Seven of eight research workers had febrile reactions 6-12 hours after exposure to SCP dust. All workers had high titres of IgG and IgM antibodies against the pseudomonas species as measured with indirect ELISA and passive haemagglutination techniques. The mechanism behind the febrile reaction is judged to be a non-immunological reaction caused by endotoxins. By increasing the particle size of the SCP through using different drying procedures, a product which generated less dust was obtained.


Predicting energy expenditures in burned patients.
Daily caloric requirements in patients with major burns are frequently estimated using the Curreri formula (25 X body weight (kg) + 40 X % BSA burned). In nonburned patients modifications of the Harris-Benedict formulas have been used to estimate energy requirements. These equations have not been validated against measured energy expenditures in burned patients. Thirty-five patients with second- and third-degree burns covering from 10 to 75% of the body surface area underwent assessments of energy expenditures by indirect calorimetry and by the Curreri and Harris-Benedict formulas. The mean energy expenditure calculated from the Harris-Benedict formulas (BEE) underestimated the mean measured energy expenditure (MEE) by 23% (p less than 0.001), while the mean energy expenditure calculated from the Curreri formula (CEE) overestimated the mean MEE by 58% (p less than 0.001). There were significant correlations between the MEE, BEE, and CEE. In patients with burns greater than 20% of the body surface area, the correlation between MEE and BEE was higher than that found with the Curreri formula.


Long-term results of radiofrequency catheter ablation in patients with Wolff-Parkinson-White syndrome.
Information about the long-term results of radiofrequency catheter ablation, electrophysiologic characteristics of differently located accessory pathways, and the difference between a single accessory pathway and multiple accessory pathways was limited. Nine hundred and thirty-one patients with 1016 accessory pathways (APs) received electrophysiologic study and radiofrequency catheter ablation between July 1, 1989 and June 31, 1996. Group 1 included 856 (91.9%) patients with a single AP and Group 2 included 75 (8.1%) patients with multiple APs. The follow-up period was 48 +/- 37 months (range, 2 to 84 months). Nine hundred and thirteen patients (98.1%) had successful ablation with a complication rate of 1.5%. In Group 1, left free wall pathways were ablated with fewer radiofrequency pulses, shorter procedure time, shorter radiation exposure time and a lower recurrence rate than those at other locations. Comparisons between Group 1 and Group 2 showed that the latter had higher incidences of antidromic tachycardia (3% vs 13%, p < 0.05) and atrial flutter/fibrillation (26% vs 37%, p < 0.05). Regarding radiofrequency catheter ablation, Group 2 needed more radiofrequency pulses (8.7 +/- 7.8 vs 5.5 +/- 7.7, p < 0.001), longer procedure time (3.3 +/- 1.4 vs 2.1 +/- 1.0 hours, p < 0.05) and radiation time (49 +/- 27 vs 29 +/- 19 minutes, p < 0.001), and a higher recurrence rate (10.6% vs 3.3%, p < 0.005) than those in Group 1. Thirty-six patients (4%) with recurrence had more right-side pathways than those without recurrence. In addition, difficult ablation (longer procedure time, longer radiation time and more radiofrequency pulses) was associated with a higher recurrence rate. These findings demonstrated that a high success rate with a low recurrence and low complication rate of radiofrequency catheter ablation could be achieved in a large population with APs during a long follow-up period.


Phenotypic and genomic comparison of Mycobacterium aurum and surrogate model species to Mycobacterium tuberculosis: implications for drug discovery.
Tuberculosis (TB) is caused by Mycobacterium tuberculosis and represents one of the major challenges facing drug discovery initiatives worldwide. The considerable rise in bacterial drug resistance in recent years has led to the need of new drugs and drug regimens. Model systems are regularly used to speed-up the drug discovery process and circumvent biosafety issues associated with manipulating M. tuberculosis. These include the use of strains such as Mycobacterium smegmatis and Mycobacterium marinum that can be handled in biosafety level 2 facilities, making high-throughput screening feasible. However, each of these model species have their own limitations. We report and describe the first complete genome sequence of Mycobacterium aurum ATCC23366, an environmental mycobacterium that can also grow in the gut of humans and animals as part of the microbiota. This species shows a comparable resistance profile to that of M. tuberculosis for several anti-TB drugs. The aims of this study were to (i) determine the drug resistance profile of a recently proposed model species, Mycobacterium aurum, strain ATCC23366, for anti-TB drug discovery as well as Mycobacterium smegmatis and Mycobacterium marinum (ii) sequence and annotate the complete genome sequence of this species obtained using Pacific Bioscience technology (iii) perform comparative genomics analyses of the various surrogate strains with M. tuberculosis (iv) discuss how the choice of the surrogate model used for drug screening can affect the drug discovery process. We describe the complete genome sequence of M. aurum, a surrogate model for anti-tuberculosis drug discovery. Most of the genes already reported to be associated with drug resistance are shared between all the surrogate strains and M. tuberculosis. We consider that M. aurum might be used in high-throughput screening for tuberculosis drug discovery. We also highly recommend the use of different model species during the drug discovery screening process.


Multifunctional nanostructured drug delivery carriers for cancer therapy: Multimodal imaging and ultrasound-induced drug release.
Development of multimodal systems for therapy and diagnosis of neoplastic diseases is an unmet need in oncology. The possibility of simultaneous diagnostics, monitoring, and therapy of various diseases allows expanding the applicability of modern systems for drug delivery. We have developed hybrid particles based on biocompatible polymers containing magnetic nanoparticles (MNPs), photoacoustic (MNPs), fluorescent (Cy5 or Cy7 dyes), and therapeutic components (doxorubicin). To achieve high loading efficiency of MNP and Dox to nanostructured carriers, we utilized a novel freezing-induced loading technique. To reduce the systemic toxicity of antitumor drugs and increase their therapeutic efficacy, we can use targeted delivery followed by the remote control of drug release using high intensity-focused ultrasound (HIFU). Loading of MNPs allowed performing magnetic targeting of the carriers and enhanced optoacoustic signal after controlled destruction of the shell and release of therapeutics as well as MRI imaging. The raster scanning optoacoustic mesoscopy (PA, RSOM), MRI, and fluorescent tomography (FT) confirmed the ultrasound-induced release of doxorubicin from capsules: in vitro (in tubes and pieces of meat) and in vivo (after delivery to the liver). Disruption of capsules results in a significant increase of doxorubicin and Cy7 fluorescence initially quenched by magnetite nanoparticles that can be used for real-time monitoring of drug release in vivo. In addition, we explicitly studied cytotoxicity, intracellular localization, and biodistribution of these particles. Elaborated drug delivery carriers have a good perspective for simultaneous imaging and focal therapy of different cancer types, including liver cancer.


[Study on the relationships between cultural orientation, alcohol expectancy, self-efficacy and drinking behavior among senior high school students in two cities of Henan province].
To explore the relationships between alcohol expectancy,cultural orientation, self-efficacy and drinking behavior to provide theoretical support for the development of education programs aimed at preventing alcohol abuse among adolescents. An anonymous quantitative survey of 2756 tenth and eleventh grade students in six senior high schools in Zhengzhou and Xinyang city of Henan province was conducted in November 2005 and data was analyzed by SPSS 13.0 software. The overall rate of drinking alcohol among participants was 56.5%. The median score of cultural orientation was 3.25 (3.09, 3.42); of alcohol expectancy was 3.02 (2.81, 3.23); and 79.64 (60.36, 93.21) of alcohol self-efficacy. Direct or indirect relationships were found between Chinese traditional cultural orientation, western cultural orientation, alcohol positive expectancy, alcohol negative expectancy, alcohol self-efficacy and alcohol drinking behavior. Lower monthly drinking habit was directly associated with higher self-efficacy and higher negative expectancies,lower positive expectancies and lower western cultural orientation (coefficients = -0.346, -0.282, 0.234 and 0.162 respectively), but not with Chinese cultural orientation variables. The current situation of alcohol drinking among the participants was critical. Drinking behavior seemed most affected by drinking beliefs and cultural orientation, suggesting that the development of education programs should focus on students' beliefs.


Detection of mild cognitive impairment in a community-dwelling population using quantitative, multiparametric MRI-based classification.
Early and accurate mild cognitive impairment (MCI) detection within a heterogeneous, nonclinical population is needed to improve care for persons at risk of developing dementia. Magnetic resonance imaging (MRI)-based classification may aid early diagnosis of MCI, but has only been applied within clinical cohorts. We aimed to determine the generalizability of MRI-based classification probability scores to detect MCI on an individual basis within a general population. To determine classification probability scores, an AD, mild-AD, and moderate-AD detection model were created with anatomical and diffusion MRI measures calculated from a clinical Alzheimer's Disease (AD) cohort and subsequently applied to a population-based cohort with 48 MCI and 617 normal aging subjects. Each model's ability to detect MCI was quantified using area under the receiver operating characteristic curve (AUC) and compared with an MCI detection model trained and applied to the population-based cohort. The AD-model and mild-AD identified MCI from controls better than chance level (AUC = 0.600, p = 0.025; AUC = 0.619, p = 0.008). In contrast, the moderate-AD-model was not able to separate MCI from normal aging (AUC = 0.567, p = 0.147). The MCI-model was able to separate MCI from controls better than chance (p = 0.014) with mean AUC values comparable with the AD-model (AUC = 0.611, p = 1.0). Within our population-based cohort, classification models detected MCI better than chance. Nevertheless, classification performance rates were moderate and may be insufficient to facilitate robust MRI-based MCI detection on an individual basis. Our data indicate that multiparametric MRI-based classification algorithms, that are effective in clinical cohorts, may not straightforwardly translate to applications in a general population.


Hierarchical nanocomposites of graphene oxide and PEGylated protoporphyrin as carriers to load doxorubicin hydrochloride for trimodal synergistic therapy.
Multimodal and synergistic therapy of cancer has appeared as one of the most promising strategies in treating cancer. Here, we report a supramolecular hierarchical nanocomposite for combination photodynamic, photothermal, and chemotherapy. Graphene oxide (GO, photothermal agent for photothermal therapy, PTT), protoporphyrin IX (PpIX, photosensitizer for photodynamic therapy, PDT), and hydrophilic anticancer drug doxorubicin hydrochloride (DOX·HCl, therapeutic for chemotherapy) are involved in hierarchical self-assembled nanocomposites via supramolecular interactions. PEGylated PpIX (PEG-PpIX) is prepared to improve the stability of GO in physiological conditions. The nanocomposite GO(PEG-PpIX) is non-cytotoxic in the dark and phototoxic with light irradiation to exhibit efficient PTT and PDT effects. The drug loading content of the nanocomposite DOX/GO(PEG-PpIX) is as high as 15.9% and the drug release shows a pH-dependent profile. The combined PDT, PTT, and chemotherapy shows an excellent in vivo antitumor effect and side effect reduction. This work presents a facile yet robust strategy to fabricate a nanocomposite for multimodal synergistic therapy.


Tuna Species Substitution in the Spanish Commercial Chain: A Knock-On Effect.
Intentional mislabelling of seafood is a widespread problem, particularly with high-value species like tuna. In this study we examine tuna mislabelling, deliberate species substitution, types of substitution and its impact on prices. The survey covered the commercial chain, from Merca-Barna to fishmongers and restaurants in the Spanish Autonomous Community of Catalonia. To understand the geographic extent of the problem we also sampled Merca-Madrid, Europe's biggest fish market, and Merca-Málaga for its proximity to the bluefin tuna migratory route and trap fishery. Monthly surveys were carried out over one year. The results showed a high deficiency in labelling: 75% of points of sale and 83% of restaurants did not specify the species, and in those cases the name of the species had to be asked. A total of 375 samples were analysed genetically, the largest dataset gathered in Europe so far. The identified species were Thunnus albacares, Thunnus thynnus and Thunnus obesus. Species substitution began at suppliers, with 40% of observed cases, increasing to 58% at fishmongers and 62% at restaurants. The substitution was mainly on bluefin tuna (T. thynnus), 73% of cases. At restaurants, only during the bluefin fishing season, we observed a decrease of Bluefin tuna substitution and an increase of reverse substitution revealing some illegal fishing. The effect of species substitution on species prices was relevant: T. obesus increased its price by around €12 kg-1 when it was sold as bluefin. In view of the deficiency of labelling, the abuse of generic names and the lack of the bluefin catch document, we conclude that the Spanish regulations are ineffective, highlighting the need for policy execution, and the urgent need for information campaigns to Spanish consumers.


First Report of Neofusicoccum australe (Botryosphaeria australis), as a Branch Dieback Pathogen of Avocado Trees in Chile.
Since 2007, Chilean avocado (Persea americana Mill.) orchards have been exposed to several abiotic stress conditions, namely frost damage and drought, due to three consecutive seasons of cold winters and shortage of irrigation water. At the same time, a severe disease resulting in tree dieback of cv. Hass, specifically, was observed in north-central Chile. Symptomatic trees exhibited abundant dead twigs in the tree canopy, and wilted leaves remained attached to the twigs in autumn. Closer inspection revealed reddish-brown necrotic lesions on the bark of the dead twigs, which girdled these symptomatic branches. When the bark was removed, the wood below appeared dark brown, in contrast to the yellowish-green coloring of healthy. The fungus was also consistently isolated from rotted fruit. A Neofusicoccum sp. with a yellow colony was consistently isolated from the necrotic lesions on PDA and incubated at room temperature for 3 days. Conidia produced in black pycnidia growing on 2% water agar with sterilized pine needles were smooth, unicellular, hyaline, and with granular contents. One or two septa developed at germination, but rarely before. The average length of the conidia was 27.0 ± 0.9 μm, with a length/width ratio of 3.9 ± 0.2 μm. Based on culture and conidial morphology, the isolates were putatively identified as Neofusicoccum luteum (1). DNA sequence analysis of the rDNA internal transcribed spacer (ITS) region was conducted for four representative isolates using primers ITS1 and ITS4 (4). The sequence analysis of ITS region of kiwifruit isolate H1M4 (Accession No. KC330230) reveled 100% nucleotide identity to N. australe (FJ157187 to FJ157192) (3). Pathogenicity tests were conducted with stem inoculations of 2-year-old cv. Hass plants grow in plastic containers in a sand/lime/peat mixture. For each inoculated plant (n = 8), a 7-mm-diameter agar plug from the margin of a 3-day culture was used as inoculum after wounding the stem to the depth to 7 mm with a cork borer. Negative control (n = 8) were wounded and then 'mock-inoculated' with sterile agar plugs. The inoculation sites were wrapped with Parafilm. All plants were kept in a greenhouse. After 5 months, all inoculated plants showed bark cankers and necrotic lesions beneath the bark, which were 5.2 cm long (n = 8). No symptoms developed on the control plants. N. australe was recovered from the margin of the necrosis lesion of every inoculated plant, thus fulfilling Koch's postulates and confirming its pathogenicity. Botryosphaeraceae spp. are the commonly reported to have ability to survive endophytically in their host, causing disease only when the host is exposed to a stress condition (2). To our knowledge, this is the first report of N. australe as a pathogen of avocado in Chile. The fungal isolates (PaHass No. 1 to 4) were deposited in the Laboratorio de Fitopatología Frutal y Molecular, Departamento de Sanidad Vegetal, Facultad de Ciencias Agronómicas de la Universidad de Chile. References: (1) A. J. L. Phillips. http://www.crem.fct.unl.pt/botryosphaeria_site/ Accessed November 20, 2011. (2) B. Slippers and M. J. Wingfield. Fungal Biol. Rev. 21:90, 2007. (3) B. Slippers et al. Mycologia 96:1030, 2004. (4) White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.


Polysaccharides in Grifola frondosa mushroom and their health promoting properties: A review.
Grifola frondosa (Dicks.) Gray is a widely consumed edible and medicinal fungus, Ancient books record that it can boost qi and fortify the spleen, moisten the lung and protect the liver. Modern people mainly use it to assist in the treatment of diabetes mellitus and various cancers. Over the past three decades, G. frondosa polysaccharides were shown to possess various promising bioactivities, mainly including antitumor and immunomodulation, anti-oxidation, anti-hyperglycemia, and meanwhile can effectively act on the skin and hematopoietic stem cells. The purpose of the present review is to provide systematically reorganized information on structural characteristics, biological activities, and structure-activity relationship of G. frondosa polysaccharides to support their further therapeutic potentials and sanitarian functions.


Personalized nanomedicine.
Personalized medicine aims to individualize chemotherapeutic interventions on the basis of ex vivo and in vivo information on patient- and disease-specific characteristics. By noninvasively visualizing how well image-guided nanomedicines-that is, submicrometer-sized drug delivery systems containing both drugs and imaging agents within a single formulation, and designed to more specifically deliver drug molecules to pathologic sites-accumulate at the target site, patients likely to respond to nanomedicine-based therapeutic interventions may be preselected. In addition, by longitudinally monitoring how well patients respond to nanomedicine-based therapeutic interventions, drug doses and treatment protocols can be individualized and optimized during follow-up. Furthermore, noninvasive imaging information on the accumulation of nanomedicine formulations in potentially endangered healthy tissues may be used to exclude patients from further treatment. Consequently, combining noninvasive imaging with tumor-targeted drug delivery seems to hold significant potential for personalizing nanomedicine-based chemotherapeutic interventions, to achieve delivery of the right drug to the right location in the right patient at the right time.


A latent profile analysis of intimate partner victimization and aggression and examination of between-class differences in psychopathology symptoms and risky behaviors.
Intimate partner violence (IPV) is associated with heightened psychopathology symptoms and risky behaviors. However, extant investigations are limited by their focus on IPV victimization, despite evidence to suggest that victimization and aggression frequently co-occur. Further, research on these correlates often has not accounted for the heterogeneity of women who experience victimization. The present study utilized latent profile analysis to identify patterns of physical, psychological, and sexual victimization and aggression in a convenience sample of 212 community women experiencing victimization (Mage = 36.63, 70.8% African American), as well as examined differences in psychopathology symptoms (i.e., posttraumatic stress symptoms [PTSS] and depressive symptoms) and risky behaviors (i.e., drug problems, alcohol problems, deliberate self-harm (DSH), HIV-risk behaviors) across these classes. Four classes of women differentiated by severities of victimization and aggression were identified. Greater psychopathology symptoms were found among classes defined by greater victimization and aggression, regardless of IPV type. Risky behaviors were more prevalent among classes defined by greater sexual victimization and aggression in particular. Findings highlight the importance of developing interventions that target the particular needs of subgroups of women who experience victimization. (PsycINFO Database Record


Malevolence, splitting, and parental ratings by borderlines.
Malevolent object relations as well as splitting have long been considered by psychodynamic theorists as central features of borderline personality disorder. We tested the hypotheses that borderlines would a) perceive their parents more negatively than both nonborderline major depressive patients and nonpatient normal controls, and b) split their representations of their parents into opposites more than the comparison subjects. Borderlines (N = 31), who were identified by the Diagnostic Interview for Borderlines, Research Diagnostic Criteria major depressives (N = 15), and nonpatient controls (N = 14) were asked to rate each parent on the Adjective Check List (ACL; Gough and Heilbrun, 1983). Seven ACL scales were studied: Favorable, Unfavorable, Critical Parent, Nurturing Parent, Nurturance, Aggression, and Dominance. Correlations were performed between scores for mother and father on the various scales for each of the three cohorts. Analysis of variance and one-way t-tests with Bonferroni correction were used to test group differences. Borderlines rated their parents, especially their fathers, not only as more unfavorable on negative scales than depressives or normals, but as less favorable on positive scales than the comparison groups. Analysis of covariance revealed that a significant portion of the variance in father scores, but not in mother scores, was related to age of respondent and history of sexual abuse. While borderlines did not appear to split their parents into one good and one bad parent, they did show significantly less correlation between parents on the Favorable scale when compared with either depressives or normal subjects. The results imply that borderlines have a greater tendency to view the world in negative, malevolent ways than to split their object representations.


Nanotechnology on duty in medical applications.
At the beginning of 21(st) century, fifty years after discovery of deoxyribonucleic acid (DNA) double helix structure, scientific world is faced with a great progress in many disciplines of biological research, especially in the field of molecular biology and operating on nucleid acid molecules. Many molecular biology techniques have been implemented successfully in biology, biotechnology, medical science, diagnostics, and many more. The introduction of polymerase chain reaction (PCR) resulted in improving old and designing new laboratory devices for PCR amplification and analysis of amplified DNA fragments. In parallel to these efforts, the nature of DNA molecules and their construction have attracted many researchers. In addition, some studies concerning mimicking living systems, as well as developing and constructing artificial nanodevices, such as biomolecular sensors and artificial cells, have been conducted. This review is focused on the potential of nanotechnology in health care and medicine, including the development of nanoparticles for diagnostic and screening purposes, the manufacture of unique drug delivery systems, antisense and gene therapy applications and the enablement of tissue engineering, including the future of nanorobot construction.


Crystal Structure and Proteomics Analysis of Empty Virus-like Particles of Cowpea Mosaic Virus.
Empty virus-like particles (eVLPs) of Cowpea mosaic virus (CPMV) are currently being utilized as reagents in various biomedical and nanotechnology applications. Here, we report the crystal structure of CPMV eVLPs determined using X-ray crystallography at 2.3 Å resolution and compare it with previously reported cryo-electron microscopy (cryo-EM) of eVLPs and virion crystal structures. Although the X-ray and cryo-EM structures of eVLPs are mostly similar, there exist significant differences at the C terminus of the small (S) subunit. The intact C terminus of the S subunit plays a critical role in enabling the efficient assembly of CPMV virions and eVLPs, but undergoes proteolysis after particle formation. In addition, we report the results of mass spectrometry-based proteomics analysis of coat protein subunits from CPMV eVLPs and virions that identify the C termini of S subunits undergo proteolytic cleavages at multiple sites instead of a single cleavage site as previously observed.


Structural and photophysical characterization of the small ultra-red fluorescent protein.
The small Ultra-Red Fluorescent Protein (smURFP) represents a new class of fluorescent protein with exceptional photostability and brightness derived from allophycocyanin in a previous directed evolution. Here, we report the smURFP crystal structure to better understand properties and enable further engineering of improved variants. We compare this structure to the structures of allophycocyanin and smURFP mutants to identify the structural origins of the molecular brightness. We then use a structure-guided approach to develop monomeric smURFP variants that fluoresce with phycocyanobilin but not biliverdin. Furthermore, we measure smURFP photophysical properties necessary for advanced imaging modalities, such as those relevant for two-photon, fluorescence lifetime, and single-molecule imaging. We observe that smURFP has the largest two-photon cross-section measured for a fluorescent protein, and that it produces more photons than organic dyes. Altogether, this study expands our understanding of the smURFP, which will inform future engineering toward optimal FPs compatible with whole organism studies.


The interpretation of computational model parameters depends on the context.
Reinforcement Learning (RL) models have revolutionized the cognitive and brain sciences, promising to explain behavior from simple conditioning to complex problem solving, to shed light on developmental and individual differences, and to anchor cognitive processes in specific brain mechanisms. However, the RL literature increasingly reveals contradictory results, which might cast doubt on these claims. We hypothesized that many contradictions arise from two commonly-held assumptions about computational model parameters that are actually often invalid: That parameters generalize between contexts (e.g. tasks, models) and that they capture interpretable (i.e. unique, distinctive) neurocognitive processes. To test this, we asked 291 participants aged 8-30 years to complete three learning tasks in one experimental session, and fitted RL models to each. We found that some parameters (exploration / decision noise) showed significant generalization: they followed similar developmental trajectories, and were reciprocally predictive between tasks. Still, generalization was significantly below the methodological ceiling. Furthermore, other parameters (learning rates, forgetting) did not show evidence of generalization, and sometimes even opposite developmental trajectories. Interpretability was low for all parameters. We conclude that the systematic study of context factors (e.g. reward stochasticity; task volatility) will be necessary to enhance the generalizability and interpretability of computational cognitive models.


Multisystem neuroprosthetic training improves bladder function after severe spinal cord injury.
Severe spinal cord injury leads to neurogenic bladder dysfunction. We recently developed a multisystem neuroprosthetic training program that promotes plastic changes capable of restoring refined locomotion in rats with severe spinal cord injury. We investigated whether multisystem neuroprosthetic training would influence the development of posttraumatic bladder dysfunction. Eight and 4 adult rats were randomly assigned to a spinal cord injury and an intact control group, respectively. Spinal cord injury consisted of 2 opposite lateral hemisections (T7 and T11), thus, interrupting all direct supraspinal input. After spinal cord injury 4 rats were subjected to a multisystem neuroprosthetic training program and 4 were not trained. At 8 weeks we performed urodynamics and evaluated kidney function using creatinine and cystatin C. Bladder investigation included morphological, histological and immunohistochemical evaluations. Bladder capacity increased threefold in trained and sevenfold in nontrained rats compared to intact rats. During filling we found a mean ± SEM of 2.7 ± 1.1 vs 12.6 ± 5.2 nonvoiding contractions in trained vs nontrained rats. Bladder morphology was similar in trained and intact rats. Nontrained rats showed detrusor hypertrophy, characterized by increased detrusor thickness and a decreased connective tissue-to-smooth muscle ratio. As labeled with protein gene product 9.5, general nerve density was significantly increased in trained and significantly decreased in nontrained rats. The relative proportion of neurofilament 200 positive afferent nerves was significantly lower in trained than in intact and nontrained rats. Neuropeptide Y positive fibers showed significantly lower density in nontrained rats. Multisystem neuroprosthetic training effectively counteracts the formation of neurogenic bladder dysfunction after severe spinal cord injury and might contribute to preserving bladder function and preventing long-term complications in patients with severe spinal cord injury.


Cardiovascular determinants of life span.
The prevalence of cardiovascular diseases rises with aging and is one of the main causes of mortality in western countries. In view of the progressively aging population, there is an urge for a better understanding of age-associated cardiovascular diseases and its underlying molecular mechanisms. The risk factors for cardiovascular diseases include unhealthy diet, diabetes, obesity, smoking, alcohol consumption, physical inactivity, and aging. Increased production of oxygen-derived free radicals plays an important role in mediating cardiovascular diseases. Oxidative stress affects the availability and/or balance of key-regulators of vascular homeostasis and favors the development of cardiovascular diseases. Reactive oxygen species are generated by different intracellular molecular pathways principally located in the cytoplasm and in the mitochondria. The mitochondrial protein p66Shc and the deacetylase enzyme SIRT1 were shown to be involved in different aspects of cardiovascular diseases. This review focuses on the latest scientific advances in understanding cardiovascular diseases associated to aging, as well as delineating the possible therapeutic implications of p66Shc and SIRT 1 in this process.


Exogenous administration of a recombinant variant of TWEAK impairs healing after myocardial infarction by aggravation of inflammation.
Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) and its receptor fibroblast growth factor-inducible 14 (Fn14) are upregulated after myocardial infarction (MI) in both humans and mice. They modulate inflammation and the extracellular matrix, and could therefore be important for healing and remodeling after MI. However, the function of TWEAK after MI remains poorly defined. Following ligation of the left coronary artery, mice were injected twice per week with a recombinant human serum albumin conjugated variant of TWEAK (HSA-Flag-TWEAK), mimicking the activity of soluble TWEAK. Treatment with HSA-Flag-TWEAK resulted in significantly increased mortality in comparison to the placebo group due to myocardial rupture. Infarct size, extracellular matrix remodeling, and apoptosis rates were not different after MI. However, HSA-Flag-TWEAK treatment increased infiltration of proinflammatory cells into the myocardium. Accordingly, depletion of neutrophils prevented cardiac ruptures without modulating all-cause mortality. Treatment of mice with HSA-Flag-TWEAK induces myocardial healing defects after experimental MI. This is mediated by an exaggerated neutrophil infiltration into the myocardium.


Visualization and visual analysis of multifaceted scientific data: a survey.
Visualization and visual analysis play important roles in exploring, analyzing, and presenting scientific data. In many disciplines, data and model scenarios are becoming multifaceted: data are often spatiotemporal and multivariate; they stem from different data sources (multimodal data), from multiple simulation runs (multirun/ensemble data), or from multiphysics simulations of interacting phenomena (multimodel data resulting from coupled simulation models). Also, data can be of different dimensionality or structured on various types of grids that need to be related or fused in the visualization. This heterogeneity of data characteristics presents new opportunities as well as technical challenges for visualization research. Visualization and interaction techniques are thus often combined with computational analysis. In this survey, we study existing methods for visualization and interactive visual analysis of multifaceted scientific data. Based on a thorough literature review, a categorization of approaches is proposed. We cover a wide range of fields and discuss to which degree the different challenges are matched with existing solutions for visualization and visual analysis. This leads to conclusions with respect to promising research directions, for instance, to pursue new solutions for multirun and multimodel data as well as techniques that support a multitude of facets.


A novel validated stability indicating high performance liquid chromatographic method for estimation of degradation behavior of ciprofloxacin and tinidazole in solid oral dosage.
The objective of current investigation was to study the degradation behavior of Ciprofloxacin and Tinidazole. The study was performed as per International Conference on Harmonization recommended stress condition. A novel stability-indicating reverse phase HPLC method was developed for the determination of Ciprofloxacin and Tinidazole purity in the presence of its impurities and forced degradation products. This method is also capable to separate placebo peaks as well in pharmaceutical dosage forms. The solid oral dosage form was subjected to the stress conditions such as oxidative, acid, base hydrolysis, heat and photolytic degradation. The method was developed using Waters symmetry shield, Reverse Phase (RP) C18, 250mm x 4.6mm, 5μ as a stationary phase. The mobile phase containing a gradient mixture of solvent A and B. 10mM phosphate buffer, adjusted pH 3.0 with phosphoric acid was used as a buffer. Buffer pH 3.0 was used as solvent A and buffer pH 3.0: Acetonitrile in the ratio of 20: 80 v/v were used as solvent B. The eluted compounds were monitored 278 nm (Ciprofloxacin), 317 nm (Tinidazole). The run time was 50 minute. In the precision study the % RSD for the result of Ciprofloxacin, Tinidazole and its impurities was below 10%. The method was linear with the correlation coefficient greater than 0.997. The percentage recoveries were calculated and observed from 93.0% to 106.7%. The peak purity of Ciprofloxacin, Tinidazole peak had not shown any flag, thus proved the stability-indicating power of the method. The developed method was validated as per ICH guidelines with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness.


Prevalence of asymptomatic incidental aneurysms: review of 4568 arteriograms.
The prevalence of unruptured cerebral aneurysms is unknown, but is estimated to be as high as 5%. The goal of this study was to determine the prevalence of asymptomatic incidental aneurysms. The authors studied all cerebral arteriography reports produced at a single institution, the University of Virginia, between April 1969 and January 1980. A review of 3684 arteriograms demonstrated 24 cases of asymptomatic aneurysms, yielding a prevalence rate of 0.65%. The majority (67%) of the 24 patients harboring unruptured aneurysms were women. More than 90% of the unruptured aneurysms were located in the anterior circulation and in locations similar to those found in patients with ruptured aneurysms. Nearly 80% of the aneurysms were smaller than 1 cm in their greatest diameter. The frequency of asymmetrical unruptured aneurysms (0.6-1.5%) was constant throughout all relevant age ranges (35-84 years). While keeping in mind appropriate caveats in extrapolating from these data, the prevalence rate of asymptomatic unruptured aneurysms found in the present study allows an estimation of the yearly rate of rupture of these lesions. The authors suggest that this yearly rate of rupture falls within the range of 1 to 2%.


Homologous liver parenchymal cell-cell adhesion mediated by an endogenous lectin and its receptor.
Many studies have implicated cell-surface lectins in heterologous cell-cell adhesion, but little is known about the participation of lectins in cellular adhesion in homologous cells. Here, we show the development of a cell model for investigating the direct role of a cell-surface lectin in homologous cell-cell adhesion. Parenchymal cells were isolated from caprine liver using a perfusion buffer, and dispersed in a chemically defined modified Ringer's solution. These cells undergo autoagglutination in the presence of Ca(2+). The autoagglutinated cells can be dissociated specifically with D-galactose (50 mM), which also inhibits the liver cell autoagglutination event. The blood serum protein fetuin has no effect on liver cell autoagglutination, whereas desialylated fetuin (100 microM), with its terminal D-galactose residue, showed a high affinity for blocking the autoagglutination event. The data demonstrates the occurrence of a Ca(2+)-dependent D-galactose-specific lectin and a lectin receptor on the parenchymal cells. Furthermore, it shows that the observed autoagglutination event is caused by the interaction of the cell-surface lectin with its receptor on the neighbouring homologous cells. The data supports the view that homologous cell-cell contact in mammalian tissues is triggered by such lectin-receptor interaction and that the previously reported cell-surface adhesive proteins serve as a secondary force to strengthen cell adhesion. This cell model could be extremely useful for investigating the direct role of cell-surface lectin and its receptor in homologous cell adhesion in a variety of tissues under normal and pathological conditions.


Voluntary consumption of morphine in 15 inbred mouse strains.
To determine genetic differences in voluntary morphine consumption, 15 commonly used inbred strains of mice were given ad libitum two-bottle choice between saccharin alone or saccharin/morphine in one bottle and water in the other bottle. Subsequently, the saccharin was gradually reduced to zero, leaving only morphine. Independent groups of mice of the same strains were exposed to quinine in a parallel manner to control for the bitter alkaloid taste of morphine. Of the 15 strains, the C57BL/6J strain showed the highest consumption of morphine, both with or without saccharin and greatest consumption of morphine relative to quinine; it also showed only a slight decline in fluid consumption when morphine was added to the saccharin bottle. In marked contrast, the SWR/J strain showed the least consumption of morphine by the same criteria, followed closely by the AKR/J, CE/J, DBA/2J and SJL/J strains. The strain differences for all the morphine drinking measures exceeded an order of magnitude. Strain-specific voluntary morphine/saccharin consumption was not significantly correlated with saccharin consumption alone, but was highly correlated with morphine consumption alone. The results show that these behaviors are under an unusually large degree of genetic determination, and some of the largest strain differences remained essentially the same regardless of whether saccharin was present, or whether quinine was used as a control tastant.


Three-dimensional organization of Drosophila melanogaster interphase nuclei. I. Tissue-specific aspects of polytene nuclear architecture.
Interphase chromosome organization in four different Drosophila melanogaster tissues, covering three to four levels of polyteny, has been analyzed. The results are based primarily on three-dimensional reconstructions from unfixed tissues using a computer-based data collection and modeling system. A characteristic organization of chromosomes in each cell type is observed, independent of polyteny, with some packing motifs common to several or all tissues and others tissue-specific. All chromosomes display a right-handed coiling chirality, despite large differences in size and degree of coiling. Conversely, in each cell type, the heterochromatic centromeric regions have a unique structure, tendency to associate, and intranuclear location. The organization of condensed nucleolar chromatin is also tissue-specific. The tightly coiled prothoracic gland chromosomes are arrayed in a similar fashion to the much larger salivary gland chromosomes described previously, having polarized orientations, nonintertwined spatial domains, and close packing of the arms of each autosome, whereas hindgut and especially the unusually straight midgut chromosomes display striking departures from these regularities. Surprisingly, gut chromosomes often appear to be broken in the centric heterochromatin. Severe deformations of midgut nuclei observed during gut contractions in living larvae may account for their unusual properties. Finally, morphometric measurements of chromosome and nuclear dimensions provide insights into chromosome growth and substructure and also suggest an unexpected parallel with diploid chromatin organization.


Pomelo II: finding differentially expressed genes.
Pomelo II (http://pomelo2.bioinfo.cnio.es) is an open-source, web-based, freely available tool for the analysis of gene (and protein) expression and tissue array data. Pomelo II implements: permutation-based tests for class comparisons (t-test, ANOVA) and regression; survival analysis using Cox model; contingency table analysis with Fisher's exact test; linear models (of which t-test and ANOVA are especial cases) that allow additional covariates for complex experimental designs and use empirical Bayes moderated statistics. Permutation-based and Cox model analysis use parallel computing, which permits taking advantage of multicore CPUs and computing clusters. Access to, and further analysis of, additional biological information and annotations (PubMed references, Gene Ontology terms, KEGG and Reactome pathways) are available either for individual genes (from clickable links in tables and figures) or sets of genes. The source code is available, allowing for extending and reusing the software. A comprehensive test suite is also available, and covers both the user interface and the numerical results. The possibility of including additional covariates, parallelization of computation, open-source availability of the code and comprehensive testing suite make Pomelo II a unique tool.


How the cell deals with DNA nicks.
During lagging strand DNA replication, the Okazaki fragment maturation machinery is required to degrade the initiator RNA with high speed and efficiency, and to generate with great accuracy a proper DNA nick for closure by DNA ligase. Several operational parameters are important in generating and maintaining a ligatable nick. These are the strand opening capacity of the lagging strand DNA polymerase delta (Pol delta ), and its ability to limit strand opening to that of a few nucleotides. In the presence of the flap endonuclease FEN1, Pol delta rapidly hands off the strand-opened product for cutting by FEN1, while in its absence, the ability of DNA polymerase delta to switch to its 3'-->5'-exonuclease domain in order to degrade back to the nick position is important in maintaining a ligatable nick. This regulatory system has a built-in redundancy so that dysfunction of one of these activities can be tolerated in the cell. However, further dysfunction leads to uncontrolled strand displacement synthesis with deleterious consequences, as is revealed by genetic studies of exonuclease-defective mutants of S. cerevisiae Pol delta. These same parameters are also important for other DNA metabolic processes, such as base excision repair, that depend on Pol delta for synthesis.


Comparing pandemic to seasonal influenza mortality: moderate impact overall but high mortality in young children.
We assessed the severity of the 2009 influenza pandemic by comparing pandemic mortality to seasonal influenza mortality. However, reported pandemic deaths were laboratory-confirmed - and thus an underestimation - whereas seasonal influenza mortality is often more inclusively estimated. For a valid comparison, our study used the same statistical methodology and data types to estimate pandemic and seasonal influenza mortality. We used data on all-cause mortality (1999-2010, 100% coverage, 16.5 million Dutch population) and influenza-like-illness (ILI) incidence (0.8% coverage). Data was aggregated by week and age category. Using generalized estimating equation regression models, we attributed mortality to influenza by associating mortality with ILI-incidence, while adjusting for annual shifts in association. We also adjusted for respiratory syncytial virus, hot/cold weather, other seasonal factors and autocorrelation. For the 2009 pandemic season, we estimated 612 (range 266-958) influenza-attributed deaths; for seasonal influenza 1,956 (range 0-3,990). 15,845 years-of-life-lost were estimated for the pandemic; for an average seasonal epidemic 17,908. For 0-4 yrs of age the number of influenza-attributed deaths during the pandemic were higher than in any seasonal epidemic; 77 deaths (range 61-93) compared to 16 deaths (range 0-45). The ≥75 yrs of age showed a far below average number of deaths. Using pneumonia/influenza and respiratory/cardiovascular instead of all-cause deaths consistently resulted in relatively low total pandemic mortality, combined with high impact in the youngest age category. The pandemic had an overall moderate impact on mortality compared to 10 preceding seasonal epidemics, with higher mortality in young children and low mortality in the elderly. This resulted in a total number of pandemic deaths far below the average for seasonal influenza, and a total number of years-of-life-lost somewhat below average. Comparing pandemic and seasonal influenza mortality as in our study will help assessing the worldwide impact of the 2009 pandemic.


Genotypic shift of the hepatitis A virus and its clinical impact on acute hepatitis A in Korea: a nationwide multicenter study.
The genotypic shift of hepatitis A virus (HAV) and its correlation with clinical course has not been evaluated in acute hepatitis A (AHA). From June 2007 to May 2009, we prospectively enrolled 546 AHA patients. We performed a nested reverse transcriptase polymerase chain reaction (RT-PCR) using the serum samples in addition to phylogenetic analysis, then we compared patient clinical features. Among 351 successfully genotyped patients, we found genotype IIIA in 178 patients (51%) and IA in 173 patients (49%). The sequences of genotype IA are identical to previously reported Korean genotype IA, and the new IIIA genotype is closely related to NOR24/Norway. We retrospectively analyzed 41 AHA samples collected from 2000 to 2006 and found that all of them were genotype IA. Patients with genotype IIIA showed significantly higher levels of aspartate aminotransferase, higher levels of alanine aminotransferase, and lower platelet counts than patients with genotype IA when comparing baseline laboratory data or peak/lowest laboratory data during the disease course. However, there were no differences in duration of hospital stay, incidence of cholestatic hepatitis, acute kidney injury, and acute liver failure, or mortality between them. A genotypic shift of the HAV was identified in Korean AHA subjects, and genotype IIIA HAV has become endemic. Although there were significant differences in the biochemical responses of AHA between genotype IA and genotype IIIA patients, we did not detect any differences in clinical outcomes such as complications or mortality.


RNAi-based gene silencing through dsRNA injection or ingestion against the African sweet potato weevil Cylas puncticollis (Coleoptera: Brentidae).
RNA interference (RNAi) technology can potentially serve as a suitable strategy to control the African sweet potato weevil Cylas puncticollis (SPW), which is a critical pest in sub-Saharan Africa. Important prerequisites are required to use RNAi in pest control, such as the presence of an efficient RNAi response and the identification of suitable target genes. Here we evaluated the toxicity of dsRNAs targeting essential genes by injection and oral feeding in SPW. In injection assays, 12 of 24 dsRNAs were as toxic as the one targeting Snf7, a gene used commercially against Diabrotica virgifera virgifera. Three dsRNAs with high insecticidal activity were then chosen for oral feeding experiments. The data confirmed that oral delivery can elicit a significant toxicity, albeit lower compared with injection. Subsequently, ex vivo assays revealed that dsRNA is affected by degradation in the SPW digestive system, possibly explaining the lower RNAi effect by oral ingestion. We conclude that the full potential of RNAi in SPW is affected by the presence of nucleases. Therefore, for future application in crop protection, it is necessary constantly to provide new dsRNA and/or protect it against possible degradation in order to obtain a higher RNAi efficacy. © 2016 Society of Chemical Industry.


Temporal Patterns of Sporulation Potential of Phomopsis viticola on Infected Grape Shoots, Canes, and Rachises.
Phomopsis cane and leaf spot on Vitis spp. (grape) is currently understood to be monocyclic, with primary inoculum only being produced early in the growing season. However, of the few published studies pertaining to sporulation of Phomopsis viticola, none specifically examined rachises, and none were designed to determine when infected tissues become capable of sporulation. The objective of these studies was to determine when grape shoots, canes, and rachises infected with P. viticola develop the capacity to sporulate, and to determine the time period during which those tissues remain capable of sporulation. Starting in 2009 and 2010, infected first-year shoots and rachises were collected biweekly throughout the growing season, into the dormant season, and into the following growing season. Tissues were collected from 'Catawba,' 'Concord,' and 'Reliance' vineyards. Samples were observed for sporulation after 48 h of incubation in a moist chamber at 23°C; the magnitude of the conidia production under these optimal conditions was considered the sporulation potential. For infections that occurred in 2009 and 2010, the production of conidia was not observed until after harvest. In the year following infection, sporulation potential was found from about bud break until shortly after the end of bloom. There was a generally consistent temporal pattern to relative sporulation potential across sampled vineyards, years, and grape tissues (rachises and canes), described by a modified β model, with peak sporulation potential occurring around 16 May. These results confirmed that Phomopsis cane and leaf spot is a monocyclic disease and support control recommendations for use of fungicides in spring.


Endogenous DNA Damage Leads to p53-Independent Deficits in Replicative Fitness in Fetal Murine Fancd2-/- Hematopoietic Stem and Progenitor Cells.
Our mechanistic understanding of Fanconi anemia (FA) pathway function in hematopoietic stem and progenitor cells (HSPCs) owes much to their role in experimentally induced DNA crosslink lesion repair. In bone marrow HSPCs, unresolved stress confers p53-dependent apoptosis and progressive cell attrition. The role of FA proteins during hematopoietic development, in the face of physiological replicative demand, remains elusive. Here, we reveal a fetal HSPC pool in Fancd2-/- mice with compromised clonogenicity and repopulation. Without experimental manipulation, fetal Fancd2-/- HSPCs spontaneously accumulate DNA strand breaks and RAD51 foci, associated with a broad transcriptional DNA-damage response, and constitutive activation of ATM as well as p38 stress kinase. Remarkably, the unresolved stress during rapid HSPC pool expansion does not trigger p53 activation and apoptosis; rather, it constrains proliferation. Collectively our studies point to a role for the FA pathway during hematopoietic development and provide a new model for studying the physiological function of FA proteins.


Long-term analysis of phase II studies of single-agent lenalidomide in relapsed/refractory mantle cell lymphoma.
Mantle cell lymphoma (MCL) is a type of non-Hodgkin lymphoma (NHL) with aggressive disease characteristics resulting in multiple relapses after initial treatment. Lenalidomide is an immunomodulatory agent approved in the US for patients with relapsed/refractory MCL following bortezomib based on results from 3 multicenter phase II studies (2 including relapsed/refractory aggressive NHL and 1 focusing on MCL post-bortezomib). The purpose of this report is to provide longer follow-up on the MCL-001 study (follow-ups were 6.8 [NHL-002], 7.6 [NHL-003], and 52.2 [MCL-001] months). The 206 relapsed MCL patients treated with single-agent lenalidomide (25 mg/day PO, days 1 to 21 every 28-days) had a median age of 67 years (63% ≥65 years), 91% with stage III/IV disease, and 50% with ≥4 previous treatment regimens. With a median follow-up of X, the combined best overall response rate (ORR) was 33% (including 11% with complete remission [CR]/CR unconfirmed CRu). Lenalidomide produced rapid and durable responses with a median time to response of 2.2 months and median duration of response (DOR) of 16.6 months (95% CI: 11.1%-29.8%). The safety profile was consistent and manageable; myelosuppression was the most common adverse event (AE). Overall, single-agent lenalidomide showed consistent efficacy and safety in multiple phase II studies of heavily pretreated patients with relapsed/refractory MCL, including those previously treated with bortezomib.


The genome of M. acetivorans reveals extensive metabolic and physiological diversity.
Methanogenesis, the biological production of methane, plays a pivotal role in the global carbon cycle and contributes significantly to global warming. The majority of methane in nature is derived from acetate. Here we report the complete genome sequence of an acetate-utilizing methanogen, Methanosarcina acetivorans C2A. Methanosarcineae are the most metabolically diverse methanogens, thrive in a broad range of environments, and are unique among the Archaea in forming complex multicellular structures. This diversity is reflected in the genome of M. acetivorans. At 5,751,492 base pairs it is by far the largest known archaeal genome. The 4524 open reading frames code for a strikingly wide and unanticipated variety of metabolic and cellular capabilities. The presence of novel methyltransferases indicates the likelihood of undiscovered natural energy sources for methanogenesis, whereas the presence of single-subunit carbon monoxide dehydrogenases raises the possibility of nonmethanogenic growth. Although motility has not been observed in any Methanosarcineae, a flagellin gene cluster and two complete chemotaxis gene clusters were identified. The availability of genetic methods, coupled with its physiological and metabolic diversity, makes M. acetivorans a powerful model organism for the study of archaeal biology. [Sequence, data, annotations and analyses are available at http://www-genome.wi.mit.edu/.]


Molecular genetic aetiology of general cognitive function is enriched in evolutionarily conserved regions.
Differences in general cognitive function have been shown to be partly heritable and to show genetic correlations with several psychiatric and physical disease states. However, to date, few single-nucleotide polymorphisms (SNPs) have demonstrated genome-wide significance, hampering efforts aimed at determining which genetic variants are most important for cognitive function and which regions drive the genetic associations between cognitive function and disease states. Here, we combine multiple large genome-wide association study (GWAS) data sets, from the CHARGE cognitive consortium (n=53 949) and UK Biobank (n=36 035), to partition the genome into 52 functional annotations and an additional 10 annotations describing tissue-specific histone marks. Using stratified linkage disequilibrium score regression we show that, in two measures of cognitive function, SNPs associated with cognitive function cluster in regions of the genome that are under evolutionary negative selective pressure. These conserved regions contained ~2.6% of the SNPs from each GWAS but accounted for ~40% of the SNP-based heritability. The results suggest that the search for causal variants associated with cognitive function, and those variants that exert a pleiotropic effect between cognitive function and health, will be facilitated by examining these enriched regions.


Magnetization transfer imaging of cortical bone in vivo using a zero echo time sequence in mice at 4.7 T: a feasibility study.
To investigate the feasibility of magnetization transfer (MT) imaging in mice in vivo for the assessment of cortical bone. MT-zero echo time data were acquired at 4.7 T in six mice using MT preparation pulses with two different flip angles (FAs) and a series of ten different off-resonance frequencies (500-15000 Hz). Regions of interest were drawn at multiple levels of the femoral cortical bone. The MT ratio (MTR) was computed for each combination of FAs and off-resonance frequencies. T1 measurements were used to estimate the direct saturation (DS) using a Bloch equation simulation. Estimation of the absorption line width of cortical bone from T2* measurements was also performed. MTR values were higher using 3000° FA than 1000° FA. MTR values decreased toward higher off-resonance frequencies. Maximum mean MTR ± standard deviation (SD) of 58.57 ± 5.22 (range 50.44-70.61) was measured with a preparation pulse of 3000° and off-resonance frequency of 500 Hz. Maximum "true" MT effect was estimated at around 2-3 and 5 kHz, respectively, for 1000° and 3000° FA. Mean full width at half maximum ± SD of 577 ± 91 Hz was calculated for the absorption spectral line of the cortical bone. MT imaging can be used for the assessment of cortical bone in mice in vivo. DS effects are negligible using preparation pulses with off-resonance frequencies greater than 3 kHz.


No changes in serum concentrations of interleukin 10 (IL-10) and interferon gamma (IF-gamma) before and after treatment of the thyroid eye disease (TED).
TED is a severe eye disease leading in rare cases to decrease of sight, optic nerve compression and blindness. Recently, significant progresses in understanding the disease have been done. Nevertheless, the treatment of the disease, especially in its severe form remains challenging. Glucocorticoids (GC) have been the basis of the treatment for a long time. Orbital irradiation (OI) and optical decompression (OD) are also used in managing the severe forms of TED. Somatostatin, intravenous immunoglobulin have been also used, with conflicting results. Regarding the potential for the treatment of TED with cytokine antagonists, controlled clinical studies are not available. Since cytokines play an important role in the pathogenesis of the TED, they seemed to be logical choice for modern TED treatment. It has been shown that both Th1 (interleukin-2, tumor necrosis factor gamma, interleukin gamma) and Th2 (interleukin -4, -5, -10) profile T cells are activated in the TED. We therefore measured interleukin-gamma, IF-gamma and interleukin -10 (IL-10)(Th1 and Th2 pattern) to assess its relationship to the course of the disease. This paper shows that both Th1 (IL-2) and Th2 (IF-gamma) pathways represented by those two cytokines are not involved (IL-10 before 2.29+/-5.23 and after treatment 3.77+/-8.44; IF gamma before 0.50+/-0.24 and after treatment 0.35+/-0.19). No relationship to the response to treatment was found. GC resulted in positive response in 8/22 patients, OI (12 patients) given after CS therapy, resulted in a response in all patients. Increase in proptosis, loss of visual acuity is spite of CS treatment prompted OD in two patients, who both recovered visual acuity and proptosis fell under 25 mm Hertel.


Proteases from Trypanosoma brucei brucei. Purification, characterisation and interactions with host regulatory molecules.
African trypanosomes contain proteases that may be released into the bloodstream of their infected hosts. This paper describes a novel, combined isolation of a cysteine proteinase (called trypanopain-Tb) and a serine oligopeptidase (which we call oligopeptidase-Tb) from Trypanosoma brucei brucei, as well as a comparison of the activities of these two enzymes against several host regulatory molecules. The enzymes differed in various respects. Firstly, purified trypanopain-Tb was shown to readily cleave proteins such as gelatin maximally at acidic pH. In contrast, oligopeptidase-Tb, which is optimally active at alkaline pH, did not hydrolyse proteins larger than 4 kDa. However, it readily hydrolysed various polypeptides, including neurotensin and atrial natriuretic factor. The interaction of the two enzymes with mammalian protease inhibitors also differed. Cystatins and alpha2-macroglobulin effectively inhibited trypanopain-Tb, with the Ki values for cystatin C and low-molecular-mass kininogen (approximately 10(-11) M) predicting, that trypanopain-Tb is likely to be effectively controlled by these inhibitors if released into the host bloodstream. In contrast, oligopeptidase-Tb was not inhibited by serpins or (a2-macroglobulin, suggesting that it may remain active if released into the host bloodstream. In support of these in vitro results, the blood of trypanosome-infected rats displayed no trypanopain-Tb-like activity, but exhibited high oligopeptidase-Tb-like activity. Thus, while trypanopain-Tb seems likely to be confined to an intracellular role within the parasite, oligopeptidase-Tb has the potential to remain active in the host bloodstream and so contribute directly to pathogenesis.


The thoracic morphology of cave-dwelling and free-living ground beetles from China (Coleoptera, Carabidae, Trechinae).
External and internal thoracic structures of two carabid species (Trechini) were examined and documented with different techniques. The study has a main focus on the eyeless cave-dwelling specialist Sinaphaenops wangorum, but detailed information is also provided for a species occurring in cave entrances. The phylogenetic background of the structural features of the thoracic skeletomuscular system was addressed. The thoracic morphology of the examined species was compared to conditions observed in previously studied carabids and non-related subterranean leiodids (Staphylinoidea) in order to identify cave adaptations. Main thoracic character complexes linked with cavernicolous habits in Trechini are elongation of the pro- and mesothorax and the legs, and a complete and irreversible reduction of the flight apparatus. The lost flight capacity is linked with a far reaching modification of skeletal elements of the metathorax including a strongly shortened and simplified metanotum, a shortened metaventrite, and completely reduced wings and sclerites of the wing base. The elongate prothorax together with the long and slender head and elongated legs distinctly increases the activity range in the subterranean lightless environment, which likely facilitates foraging of the carnivorous beetles. Some of the observed features like wing loss and elongation of the anterior thorax and legs are also found in some cave-dwelling Leiodidae (Leptodirini), whereas some other subterranean members of the staphylinoid family have a compact body and legs of normal length. In contrast to the predaceous Trechini, Leptodirini are scavengers.


Minozac treatment prevents increased seizure susceptibility in a mouse "two-hit" model of closed skull traumatic brain injury and electroconvulsive shock-induced seizures.
The mechanisms linking traumatic brain injury (TBI) to post-traumatic epilepsy (PTE) are not known and no therapy for prevention of PTE is available. We used a mouse closed-skull midline impact model to test the hypotheses that TBI increases susceptibility to seizures in a "two-hit" injury model, and that suppression of cytokine upregulation after the first hit will attenuate the increased susceptibility to the second neurological insult. Adult male CD-1 mice underwent midline closed skull pneumatic impact. At 3 and 6 h after impact or sham procedure, the mice were injected IP with either Minozac (Mzc), a suppressor of proinflammatory cytokine upregulation, or vehicle (saline). On day 7 after sham operation or TBI, seizures were induced using electroconvulsive shock (ECS), and susceptibility to seizures was measured by the current required for seizure induction. Activation of glia, neuronal injury, and metallothionein-immunoreactive cells were quantified in the hippocampus by immunohistochemical methods. Neurobehavioral function over 14-day recovery was quantified using the Barnes maze. Following TBI there was a significant increase in susceptibility to seizures induced by ECS, and this susceptibility was prevented by suppression of cytokine upregulation with Mzc. Astrocyte activation, metallothionein expression, and neurobehavioral impairment were also increased in the two-hit group subjected to combined TBI and ECS. These enhanced responses in the two-hit group were also prevented by suppression of proinflammatory cytokine upregulation with Mzc. These data implicate glial activation in the mechanisms of epileptogenesis after TBI, and identify a potential therapeutic approach to attenuate the delayed neurological sequelae of TBI.


Nuclear receptor-binding sites of coactivators glucocorticoid receptor interacting protein 1 (GRIP1) and steroid receptor coactivator 1 (SRC-1): multiple motifs with different binding specificities.
The activity of the AF-2 transcriptional activation function of nuclear receptors (NR) is mediated by the partially homologous transcriptional coactivators, glucocorticoid receptor interacting protein 1 (GRIP1)/transcriptional intermediary factor 2 (TIF2) and steroid receptor coactivator 1 (SRC-1). GRIP1 and SRC-1 bound nine different NRs and exhibited similar, but not identical, NR binding preferences. The most striking difference was seen with the androgen receptor, which bound well to GRIP1 but poorly to SRC-1. GRIP1 and SRC-1 contain three copies of the NR binding motif LXXLL (called an NR Box) in their central regions. Mutation of both NR Box II and NR Box III in GRIP1 almost completely eliminated functional and binding interactions with NRs, indicating that these two sites are crucial for most of GRIP1's NR binding activity. Interactions of GRIP1 with the estrogen receptor were more strongly affected by mutations in NR Box II, whereas interactions with the androgen receptor and glucocorticoid receptor were more strongly affected by NR Box III mutations. One isoform of SRC-1 has an additional NR Box (NR Box IV) at its extreme C terminus with an NR-binding preference somewhat different from that of the central NR-binding domain of SRC-1. GRIP1 has no NR Box in its C-terminal region and therefore no C-terminal NR-binding function. In summary, GRIP1 and SRC-1 have overlapping NR-binding preferences, but specific NRs display both coactivator and NR Box preferences that may contribute to the specificity of hormonal responses.


Joint EANM/EANO/RANO practice guidelines/SNMMI procedure standards for imaging of gliomas using PET with radiolabelled amino acids and [18F]FDG: version 1.0.
These joint practice guidelines, or procedure standards, were developed collaboratively by the European Association of Nuclear Medicine (EANM), the Society of Nuclear Medicine and Molecular Imaging (SNMMI), the European Association of Neurooncology (EANO), and the working group for Response Assessment in Neurooncology with PET (PET-RANO). Brain PET imaging is being increasingly used to supplement MRI in the clinical management of glioma. The aim of these standards/guidelines is to assist nuclear medicine practitioners in recommending, performing, interpreting and reporting the results of brain PET imaging in patients with glioma to achieve a high-quality imaging standard for PET using FDG and the radiolabelled amino acids MET, FET and FDOPA. This will help promote the appropriate use of PET imaging and contribute to evidence-based medicine that may improve the diagnostic impact of this technique in neurooncological practice. The present document replaces a former version of the guidelines published in 2006 (Vander Borght et al. Eur J Nucl Med Mol Imaging. 33:1374-80, 2006), and supplements a recent evidence-based recommendation by the PET-RANO working group and EANO on the clinical use of PET imaging in patients with glioma (Albert et al. Neuro Oncol. 18:1199-208, 2016). The information provided should be taken in the context of local conditions and regulations.


Comparison of the NO and HNO donating properties of diazeniumdiolates: primary amine adducts release HNO in Vivo.
Diazeniumdiolates, more commonly referred to as NONOates, have been extremely useful in the investigation of the biological effects of nitric oxide (NO) and related nitrogen oxides. The NONOate Angeli's salt (Na(2)N(2)O(3)) releases nitroxyl (HNO) under physiological conditions and exhibits unique cardiovascular features (i.e., positive inotropy/lusitropy) that may have relevance for pharmacological treatment of heart failure. In the search for new, organic-based compounds that release HNO, we examined isopropylamine NONOate (IPA/NO; Na[(CH(3))(2)CHNH(N(O)NO]), which is an adduct of NO and a primary amine. The chemical and pharmacological properties of IPA/NO were compared to those of Angeli's salt and a NO-producing NONOate, DEA/NO (Na[Et(2)NN(O)NO]), which is a secondary amine adduct. Under physiological conditions IPA/NO exhibited all the markers of HNO production (e.g., reductive nitrosylation, thiol reactivity, positive inotropy). These data suggest that primary amine NONOates may be useful as HNO donors in complement to the existing series of secondary amine NONOates, which are well-characterized NO donors.


Neuronal activity of the supplementary motor area (SMA) during internally and externally triggered wrist movements.
The activity of neurones was recorded from the supplementary motor area (SMA) of monkeys while they were performing a discrete, arbitrary wrist movement. The cells responded similarly whether there was a triggering stimulus at the time of the movement or not. This experiment indicates that SMA neurones are active both in relation to externally triggered and internally initiated (voluntary) actions.


Correlation of expression of Wnt-1 in developing limbs with abnormalities in growth and skeletal patterning.
The Wnt genes are members of a family of vertebrate genes related to the Drosophila gene wingless (wg). They encode secreted molecules that are thought to be important in patterning and growth control during ontogenesis. Several such genes are transcribed in localized domains during limb budding and morphogenesis. We report here a congenital limb malformation in a mouse transgenic line that ectopically expresses Wnt-1 in the developing limbs. The hemizygote phenotype, which is inherited as an autosomal dominant trait, presents extensive distal truncations of skeletal elements, skeletal fusions and interdigital webbing. The data shown here demonstrate that abnormal Wnt-1 expression is correlated with retarded mesenchymal condensations replaced by highly proliferative cells in the limb bud. This seems to lead to an inability of the affected cells to participate in normal skeletal development leading to the adult defects.


The Applicability and Performance of Tools Used to Assess the Father-Offspring Relationship in Relation to Parental Psychopathology and Offspring Outcomes.
Introduction: Father-infant interactions are important for optimal offspring outcomes. Moreover, paternal perinatal psychopathology is associated with psychological and developmental disturbances in the offspring, and this risk may increase when both parents are unwell. While, the father-offspring relationship is a plausible mechanism of risk transmission, there is presently no "gold standard" tool for assessing the father-offspring relationship. Therefore, we systematically searched and reviewed the application and performance of tools used to assess the father-offspring relationship from pregnancy to 24-months postnatal. Methods: Four electronic databases (including MEDLINE, PsycINFO, Maternity and Infant Care Database, and CINAHL) were searched. Selected articles included evidence of father-offspring relationship assessment in relation to parental perinatal psychopathology and/or offspring outcomes. Data was extracted and synthesized according to the following: (i) evidence supporting the performance of tools in terms of their psychometric properties when applied in the context of fathers, (ii) tool specific characteristics, and (iii) study specific methodological aspects in which the tool was embedded. Results: Of the 30,500 records eligible for screening, 38 unique tools used to assess the father-offspring relationship were identified, from 61 studies. Ten tools were employed in the context of paternal psychopathology, three in the context of maternal psychopathology, and seven in the context of both maternal and paternal psychopathology, while nine tools were applied in the context of offspring outcomes only. The remaining nine tools were used in the context of both parental psychopathology (i.e., paternal, and/or maternal psychopathology) and offspring outcomes. Evidence supporting the psychometric robustness of the extracted observational, self-report and interview-based tools was generally limited. Most tools were originally developed in maternal samples-with few tools demonstrating evidence of content validation specific to fathers. Furthermore, various elements influencing tool performance were recognized-including variation in tool characteristics (e.g., relationship dimensions assessed, assessment mode, and scoring formats) and study specific methodological aspects, (e.g., setting and study design, sample characteristics, timing and nature of parental psychopathology, and offspring outcomes). Conclusion: Given the strengths and limitations of each mode of assessment, future studies may benefit from a multimethod approach to assessing the father-offspring relationship, which may provide a more accurate assessment than one method alone.


Interrater reliability of identifying indicators of posterior ligamentous complex disruption when plain films are indeterminate in thoracolumbar injuries.
The Spine Trauma Study Group (STSG) has proposed a novel thoracolumbar injury classification system and score (TLICS) in an attempt to define traumatic spinal injuries and direct appropriate management schemes objectively. The TLICS assigns specific point values based on three variables to generate a final severity score that guides potential treatment options. Within this algorithm, significant emphasis has been placed on posterior ligamentous complex (PLC) integrity. The purpose of this study was to determine the interrater reliability of indicators surgeons use when assessing PLC disruption on imaging studies, including computed tomography (CT) and magnetic resonance imaging (MRI). Orthopedic surgeons and neurosurgeons retrospectively reviewed a series of thoracolumbar injury case studies. Thirteen case studies, including images, were distributed to STSG members for individual, independent evaluation of the following three criteria: (1) diastasis of the facet joints on CT; (2) posterior edema-like signal in the region of PLC components on sagittal T2-weighted fat saturation (FAT SAT) MRI; and (3) disrupted PLC components on sagittal T1-weighted MRI. Interrater agreement on the presence or absence of each of the three criteria in each of the 13 cases was assessed. Absolute interrater percent agreement on diastasis of the facet joints on CT and posterior edema-like signal in the region of PLC components on sagittal T2-weighted FAT SAT MRI was similar (agreement 70.5%). Interrater agreement on disrupted PLC components on sagittal T1-weighted MRI was 48.9%. Facet joint diastasis on CT was the most reliable indicator of PLC disruption as assessed by both Cohen's kappa (kappa = 0.395) and intraclass correlation coefficient (ICC 0.430). The interrater reliability of assessing diastasis of the facet joints on CT had fair to moderate agreement. The reliability of assessing the posterior edema-like signal in the region of PLC components was lower but also fair, whereas the reliability of identifying disrupted PLC components was poor.


Toddler Temperament Mediates the Effect of Prenatal Maternal Stress on Childhood Anxiety Symptomatology: The QF2011 Queensland Flood Study.
It is not known whether alterations to temperamental characteristics associated with prenatal maternal stress (PNMS) exposure account for the development of childhood anxiety symptomatology (internalizing behaviors and anxiety symptoms). The QF2011 Queensland flood study examined whether (1) toddler temperamental characteristics explained the association between PNMS exposure and childhood anxiety symptomatology; and (2) whether effects were dependent upon child sex or the timing of gestational exposure to PNMS. We investigated the effects of various aspects of flood-related stress in pregnancy (objective hardship, cognitive appraisal, subjective distress) on maternal report of 16-month toddler temperament (attentional control, shy-inhibition, negative reactivity), 4-year maternal-reported childhood anxiety symptomatology (internalizing and anxiety symptoms; N = 104), and teacher reports of internalizing behaviors (N = 77). Severity of maternal objective hardship during pregnancy and shy-inhibited behaviors were uniquely associated with 4-year child anxiety symptoms. Mediation analyses found that higher levels of 16-month negative reactivity accounted, in part, for the relationship between increased maternal objective flood-related hardship and greater internalizing behaviors (maternal but not teacher report). Neither child sex nor gestational timing of exposure moderated the hypothesized mediations. Our findings highlight several pathways through which varying aspects of disaster-related PNMS may influence early childhood anxiety symptomatology.


Unprecedented Arctic ozone loss in 2011.
Chemical ozone destruction occurs over both polar regions in local winter-spring. In the Antarctic, essentially complete removal of lower-stratospheric ozone currently results in an ozone hole every year, whereas in the Arctic, ozone loss is highly variable and has until now been much more limited. Here we demonstrate that chemical ozone destruction over the Arctic in early 2011 was--for the first time in the observational record--comparable to that in the Antarctic ozone hole. Unusually long-lasting cold conditions in the Arctic lower stratosphere led to persistent enhancement in ozone-destroying forms of chlorine and to unprecedented ozone loss, which exceeded 80 per cent over 18-20 kilometres altitude. Our results show that Arctic ozone holes are possible even with temperatures much milder than those in the Antarctic. We cannot at present predict when such severe Arctic ozone depletion may be matched or exceeded.


Engineering of Isogenic Cells Deficient for MR1 with a CRISPR/Cas9 Lentiviral System: Tools To Study Microbial Antigen Processing and Presentation to Human MR1-Restricted T Cells.
The nonclassical HLA molecule MHC-related protein 1 (MR1) presents metabolites of the vitamin B synthesis pathways to mucosal-associated invariant T (MAIT) cells and other MR1-restricted T cells. This new class of Ags represents a variation on the classical paradigm of self/non-self discrimination because these T cells are activated through their TCR by small organic compounds generated during microbial vitamin B2 synthesis. Beyond the fundamental significance, the invariant nature of MR1 across the human population is a tantalizing feature for the potential development of universal immune therapeutic and diagnostic tools. However, many aspects of MR1 Ag presentation and MR1-restricted T cell biology remain unknown, and the ubiquitous expression of MR1 across tissues and cell lines can be a confounding factor for experimental purposes. In this study, we report the development of a novel CRISPR/Cas9 genome editing lentiviral system and its use to efficiently disrupt MR1 expression in A459, THP-1, and K562 cell lines. We generated isogenic MR1(-/-) clonal derivatives of the A549 lung carcinoma and THP-1 monocytic cell lines and used these to study T cell responses to intracellular pathogens. We confirmed that MAIT cell clones were unable to respond to MR1(-/-) clones infected with bacteria whereas Ag presentation by classical and other nonclassical HLAs was unaffected. This system represents a robust and efficient method to disrupt the expression of MR1 and should facilitate investigations into the processing and presentation of MR1 Ags as well as into the biology of MAIT cells.


Differential pressor and renal vascular reactivity to angiotensin II in spontaneously hypertensive and Wistar-Kyoto rats.
The suggestion has been made that the Okamoto strain of spontaneously hypertensive rats (SHR) shares some features with a subgroup of patients with essential hypertension, called nonmodulators. One feature of nonmodulators is a renal blood flow response to angiotensin II (ANG II) that is blunted on a high salt diet; the blunted renal vascular response is corrected by converting enzyme inhibition. Renal blood flow (electromagnetic flowmeter) and pressor responses to graded ANG II doses (5-300 ng) were assessed in 24 SHR and 24 Wistar-Kyoto rats (WKY) ingesting 1.6% Na. In comparison to WKY, blood pressure was higher in SHR (155 +/- 4 vs 106 +/- 2 mm Hg; p less than 0.001), renal blood flow was lower (6.9 +/- 0.5 vs 8.2 +/- 0.4 ml/min/g; p less than 0.05), and the pressor response to ANG II was enhanced, (p less than 0.0005) but the renal vascular response was blunted (p less than 0.005). Captopril (1-30 mg/kg) reduced blood pressure more in SHR than in WKY but increased renal blood flow similarly in both strains. The blunted renal vascular response to ANG II in SHR was reversed by captopril, but inhibition of converting enzyme in the kidney did not parallel systemic inhibition. Maximum blockade of converting enzyme in the kidney appears to require a larger captopril dose than is required for systemic inhibition. These results suggest that the renal blood supply in SHR also shares some of the characteristics of nonmodulators and that the action of captopril on the renal blood flow probably reflects reversal of inappropriate intrarenal ANG II formation.


Antihypertensive effect of salt-free soy sauce, a new fermented seasoning, in spontaneously hypertensive rats.
In this study, we focused on the antihypertensive effect of newly fermented salt-free soy sauce (SFS) in spontaneously hypertensive rats (SHRs). A 13-wk oral administration of SFS was performed daily in 8- to 21-wk-old SHRs with a dose of 200 mg/kg of body weight/d. The systolic (SBP) and diastolic (DBP) blood pressures were measured for each week during the experimental period. As a result, a significant (P < 0.01) BP lowering effect was observed from the 5th d to the end of the experiment in the SFS group compared with the control group (DeltaSBP(21-wk): 27 mmHg, DeltaDBP(21-wk): 20 mmHg). In contrast to such substantial effect by SFS intake, angiotensin I-converting enzyme activities in blood and local organs as well as pharmaceutical parameters such as serum Na(+) or K(+) level did not show any significant difference between in both SFS and control groups. Vasoconstriction experiment using thoracic aorta rings from 21-wk SHRs provided an interesting result that the rings from the SFS group evoked a >2-fold higher increase in the angiotensin II-stimulated constrictive response compared with the rings from the control group (P = 0.012), which suggested that the SFS-intake would be effective in possessing a higher vessel tone. In this study, we investigated the antihypertensive effect of newly fermented SFS in SHRs. As a result of 13-wk SFS-administration, we found a significant antihypertensive effect of the SFS. This finding strongly demonstrates that the developed SFS would be greatly beneficial for health and useful for health-related industries.


Getting the shots: methods to gain adherence to a multi-dose vaccination program for inner city, drug-involved prostitution communities.
Street-based sex-work and poly-substance drug use, coupled with low vaccination rates and limited utilization of the mainstream health care system, put the sex worker communities of Bogotá's city center at extreme risk of infection with the hepatitis B virus (HBV). Vaccination is critical to maintaining low prevalence of the disease and low incidence of new cases, yet the floating and inconsistent nature of Bogotá's drug-involved female and transsexual prostitution communities make it difficult to complete a 3-dose vaccination program. Between December 2011 and March of 2012, the Fénix Foundation collaborated with the Bogotá Health Department to deliver free HBV vaccines to this vulnerable population. This paper outlines methods used in the vaccination program to generate a 37.7% adherence rate, significantly higher than that previously reported for HBV vaccination programs also targeting marginalized populations. This program's practices are based on the Fénix peer leader method, and are offered as a model that can be applied to other health interventions operating in analogous contexts, with similarly high-risk populations.