Internal dose escalation is associated with increased local control for non-small cell lung cancer (NSCLC) brain metastases treated with stereotactic radiosurgery (SRS).
To identify potentially actionable dosimetric predictors of local control (LC) for non-small cell lung cancer (NSCLC) brain metastases treated with single-fraction stereotactic radiosurgery (SRS). Patients with NSCLC brain metastases treated with single-fraction SRS were identified. Eligible patients had at least 1 follow-up magnetic resonance imaging scan and were without prior metastasectomy or SRS to the same lesion. LC and overall survival (OS) were estimated using the Kaplan-Meier method. The Cox proportional hazards model was used for univariate (UVA) and multivariate analysis (MVA). Receiver operating characteristic (ROC) analysis was used to identify optimal cut points for dose-volume histogram metrics relative to LC. A total of 612 NSCLC brain metastasis were identified in 299 patients with single-fraction SRS between 1999 and 2014. Median follow-up was 10 months. Median OS from time of SRS was 11 months. Overall LC was 75% and 66% at 1 and 2 years, respectively. On UVA, increasing dose by any measure was associated with improved LC. On MVA, volume receiving at least 32 Gy (V32; hazard ratio [HR], 0.069; P < .000), along with higher prescription isodose (HR, 0.953; P = .031) and lower volume (HR, 1.359; P < .000), were independent predictors of improved LC. ROC analysis demonstrated a V32 of 24% to be most predictive for LC. For the entire cohort, 1-year LC for V32 ≥24% was 89% versus 67% for V32 <24% (P = .000). Stratifying by volume, lesions ≤2 cm (n = 323) had a 1-year LC of 95% versus 82% (P = .005) for V32 above and below 24%, respectively. For lesions 2.1 to 3 cm (n = 211), 1-year LC was 79% versus 59% (P = .003) for V32 above and below 24%, respectively. Total tumor volume alone was predictive for OS. Volume, prescription isodose line, and V32 are independent predictors of LC. V32 represents an actionable SRS treatment planning parameter for NSCLC brain metastases.


Peptidic biofunctionalization of laser patterned dental zirconia: A biochemical-topographical approach.
A dual approach employing peptidic biofunctionalization and laser micro-patterns on dental zirconia was explored, with the aim of providing a flexible tool to improve tissue integration of restorations. Direct laser interference patterning with a femtosecond Ti:Sapphire laser was employed, and two periodic grooved patterns were produced with a periodicity of 3 and 10 μm. A platform containing the cell-adhesive RGD and the osteogenic DWIVA peptides was used to functionalize the grooved surfaces. Topography and surface damage were characterized by confocal laser scanning (CLSM), scanning electron and scanning transmission electron microscopy techniques. The surface patterns exhibited a high homogeneity and subsurface damage was found in the form of nano-cracks and nano-pores, at the bottom of the valleys. Accelerated tests in water steam were carried out to assess hydrothermal degradation resistance, which slightly decreased after the laser treatment. Interestingly, the detrimental effects of the laser modification were reverted by a post-laser thermal treatment. The attachment of the molecule was verified trough fluorescence CLSM and X-ray photoelectron spectroscopy. Finally, the biological properties of the surfaces were studied in human mesenchymal stem cells. Cell adhesion, morphology, migration and differentiation were investigated. Cells on grooved surfaces displayed an elongated morphology and aligned along the patterns. On these surfaces, migration was greatly enhanced along the grooves, but also highly restricted in the perpendicular direction as compared to flat specimens. After biofunctionalization, cell number and cell area increased and well-developed cell cytoskeletons were observed. However, no effects on cell migration were found for the peptidic platform. Although some osteogenic potential was found in specimens grooved with a periodicity of 10 μm, the largest effects were observed from the biomolecule, which favored upregulation of several genes related to osteoblastic differentiation in all the surfaces.


Quercetin-imprinted polymer for anthocyanin extraction from mangosteen pericarp.
Molecular imprinting is a facilitative technology for the production of artificial receptors possessing great endurance with high specificity toward target molecules of interest. The polymers are commonly applied for separation or analysis of substances of interest. In this study, we prepared molecularly imprinted polymers for the purpose of binding specifically to quercetin and related compounds. Quercetin was used as the template molecule, 4-vinylpyridine (4-VP) as the functional monomer, ethylene glycol dimethacrylate (EDMA) as the cross-linking monomer, azobisisobutyronitrile (AIBN) as the polymerization initiator and ethanol as the porogenic solvent. Such 4-VP-based imprinted polymer was found to bind the template molecule greater than that of the control polymer with an approximate 2 folds higher binding using 20mg of polymer in the optimal solvent, ethanol:water (4:1v/v). Quercetin-imprinted polymer (QIP) was found to bind well against its template; approximately 1mg/g polymer. In addition, QIP was applied to bind anthocyanin from the crude extract of mangosteen pericarp. The binding capacity of quercetin-MIP toward anthocyanin was approximately 0.875mg per gram of polymer. This result indicated that quercetin-MIP showed its specific binding to quercetin and related compound particularly anthocyanin. In conclusion, we have demonstrated the successful preparation and utilization of molecularly imprinted polymer for the specific recognition of quercetin as well as structurally related anthocyanins from the mangosteen pericarp with enhanced and robust performance.


BAD TIMES, SLIMMER CHILDREN?
Although the majority of the literature has confirmed that recessions are beneficial for adults' health and babies' outcomes at delivery, this effect should not necessarily be the same for children. In this paper, we study the effect of business cycle conditions on infant underweight, overweight, and obesity. We exploit eight waves of repeated cross-sectional data (1987-2012) of the Spanish National Health Survey for children aged 2-15 and use the regional unemployment rate of the trimester of the interview as a proxy for the business cycle phase at the local level. We find that an increase in the unemployment rate is associated with lower obesity incidence, especially for children under 6 years old and over 12 years old. However, economic shocks also proof to have potentially negative consequences as they increase the prevalence of infant underweight for the same age groups. Moreover, we show that the possible mechanisms through which the cycle is impacting infant obesity is the nutritional composition of the children's diet, as well as, increases in the frequency of exercise. We provide some evidence that suggests that the impact of business cycle conditions on infant weight disorders have little objective health consequences in the short run. However, the potential long-term effects may become important as underweight during childhood is associated with worse outcomes later in life. Copyright © 2016 John Wiley & Sons, Ltd.


The Complete Chloroplast Genome of the Vulnerable Oreocharis esquirolii (Gesneriaceae): Structural Features, Comparative and Phylogenetic Analysis.
Oreocharis esquirolii, a member of Gesneriaceae, is known as Thamnocharis esquirolii, which has been regarded a synonym of the former. The species is endemic to Guizhou, southwestern China, and is evaluated as vulnerable (VU) under the International Union for Conservation of Nature (IUCN) criteria. Until now, the sequence and genome information of O. esquirolii remains unknown. In this study, we assembled and characterized the complete chloroplast (cp) genome of O. esquirolii using Illumina sequencing data for the first time. The total length of the cp genome was 154,069 bp with a typical quadripartite structure consisting of a pair of inverted repeats (IRs) of 25,392 bp separated by a large single copy region (LSC) of 85,156 bp and a small single copy region (SSC) of18,129 bp. The genome comprised 114 unique genes with 80 protein-coding genes, 30 tRNA genes, and four rRNA genes. Thirty-one repeat sequences and 74 simple sequence repeats (SSRs) were identified. Genome alignment across five plastid genomes of Gesneriaceae indicated a high sequence similarity. Four highly variable sites (rps16-trnQ, trnS-trnG, ndhF-rpl32, and ycf 1) were identified. Phylogenetic analysis indicated that O. esquirolii grouped together with O. mileensis, supporting resurrection of the name Oreocharis esquirolii from Thamnocharisesquirolii. The complete cp genome sequence will contribute to further studies in molecular identification, genetic diversity, and phylogeny.


Semisynthetic murine prion protein equipped with a GPI anchor mimic incorporates into cellular membranes.
Conversion of cellular prion protein (PrP(C)) into the pathological conformer (PrP(Sc)) has been studied extensively by using recombinantly expressed PrP (rPrP). However, due to inherent difficulties of expressing and purifying posttranslationally modified rPrP variants, only a limited amount of data is available for membrane-associated PrP and its behavior in vitro and in vivo. Here, we present an alternative route to access lipidated mouse rPrP (rPrP(Palm)) via two semisynthetic strategies. These rPrP variants studied by a variety of in vitro methods exhibited a high affinity for liposomes and a lower tendency for aggregation than rPrP. In vivo studies demonstrated that double-lipidated rPrP is efficiently taken up into the membranes of mouse neuronal and human epithelial kidney cells. These latter results enable experiments on the cellular level to elucidate the mechanism and site of PrP-PrP(Sc) conversion.


The effect of polyvinyl pyrrolidone on the clinical activity of 0.09% and 0.2% chlorhexidine mouthrinses.
Previous studies have shown that polyvinyl pyrrolidone (PVP) added to a chlorhexidine rinse reduced extrinsic dental stain but at the expense of a reduction in plaque inhibitory activity. This effect appeared due to a reduction in the effective chlorhexidine dose to levels where dose response studies show plaque inhibition falls off rapidly. The aim of these 2 clinical studies was to determine if PVP could be added to chlorhexidine rinses to maintain efficacy and reduce staining. Study 1 involved 42 healthy dentate volunteers and was a blind, randomised, 7 treatment, crossover design balanced for residual effects. The rinses were: 1. 0.09% chlorhexidine to which was added, 2. 1% PVP, 3. 3% PVP, 4. 5% PVP, 5. 7% PVP, 6. Placebo, 7. Essential oil product. Rinses were used 2x on day one of each period after a prophylaxis. Subjects suspended tooth cleaning for 24 h and were then scored for plaque area. Study 2 used the experimental gingivitis model, involved 24 healthy dentate subjects and was a blind, randomised, 3 treatment, crossover design balanced for residual effects. The rinses were 1. 0.2% chlorhexidine, 2. 0.2% chlorhexidine/10% PVP, 3. Placebo. At baseline and the end of each study period subjects were rendered plaque, stain and calculus free, suspended oral hygiene and rinsed 2x per day. Plaque, gingivitis and stain were scored at baseline, 1, 2, and 3 weeks. Calculus was scored at baseline and 3 weeks. Study 1: Buccal plaque scores were significantly lower with all rinses compared to placebo. Also all buccal plaque scores were significantly lower with chlorhexidine and chlorhexidine/PVP rinses compared to the essential oil/phenolic rinse. There were no significant differences between the chlorhexidine rinse and the chlorhexidine/PVP rinses. Analyses for buccal and lingual plaque combined produced, with one exception, the same results for rinse comparisons as for buccal plaque alone. Thus the essential oil/phenolic rinse just failed to reach significance compared to placebo. Study 2: Plaque and gingivitis scores were significantly lower with positive control and test rinses compared to placebo but with no difference between these rinses. Tooth and tongue stain was significantly higher with the positive control and test rinses compared to placebo but not significantly different between these 2 rinses. Calculus scores were not significantly different between the three study rinses. Taken with previous data, the balance of evidence does not support PVP as an inhibitor of staining associated with chlorhexidine. These data are further evidence that chlorhexidine oral hygiene products, which, do not or claim not to cause staining, are most probably lacking efficacy.


Medical cannabis for inflammatory bowel disease: real-life experience of mode of consumption and assessment of side-effects.
Use of medical cannabis for improving symptoms of inflammatory bowel disease is increasing. However, reports on long-term outcomes are lacking. This prospective, observational study assessed the effects of licensed cannabis use among patients with inflammatory bowel disease. Dose and mode of consumption, adverse events, use of other medications, and long-term effects were evaluated among 127 patients with inflammatory bowel disease using legalized medical cannabis. Blood count, albumin, and C-reactive protein were assessed before, 1 month, and at least 1 year after medical cannabis therapy was initiated. Questionnaires on disease activity, patient function, and signs of addiction were completed by patients and by a significant family member to assess its effects. The average dose used was 31 ± 15 g/month. The average Harvey-Bradshaw index improved from 14 ± 6.7 to 7 ± 4.7 (P < 0.001) during a median follow-up of 44 months (interquartile range, 24-56 months). There was a slight, but statistically significant, average weight gain of 2 kg within 1 year of cannabis use. The need for other medications was significantly reduced. Employment among patients increased from 65 to 74% (P < 0.05). We conclude that the majority of inflammatory bowel disease patients using cannabis are satisfied with a dose of 30 g/month. We did not observe negative effects of cannabis use on the patients' social or occupational status. Cannabis use by inflammatory bowel disease patients can induce clinical improvement and is associated with reduced use of medication and slight weight gain. Most patients respond well to a dose of 30 g/month, or 21 mg Δ9-tetra- hydrocannabinol (THC) and 170 mg Cannabidiol (CBD) per day.


Something more to say about calcium homeostasis: the role of vitamin K2 in vascular calcification and osteoporosis.
Vascular calcification and osteoporosis share similar etiopathogenetic mechanisms. Vitamin K2 deficiency could be responsible of the so called "calcium paradox", that is the lack of calcium in the bone and its storage in the vessel wall. These events may have clinically relevant consequences, such as cardiovascular accidents, and bone fractures. To review the biological function of vitamin K2 metabolism, the main factors related to its deficiency and the consequent clinical significance. Vitamin K2 is essential for the function of several proteins, involved in the maintenance of the normal structure of arterial wall, osteoarticular system, teeth, and for the regulation of cell growth. It has been demonstrated to have a pivotal role in the inhibition of vascular foci of calcification, and in the regulation of calcium deposition in the bone. Vitamin K2 deficiency is often subclinic in a large part of healthy population. This deficiency is related to the interaction of various factors, such as the reduced dietary intake, the alteration of intestinal absorption or production, with a possible role of intestinal microbiota and the increased consumption at the vessel wall. Vitamin K2 deficiency has recently been recognized as a protagonist in the development of vascular calcification and osteoporosis. Data reported so far are promising and, dietary supplementation seems a useful tool to contrast these diseases. However, large studies or solid clinical correlations regarding vitamin K2 deficiency and its pathologic consequences are needed to confirm these preliminary experiences.


Chronotypes and subjective sleep parameters in epilepsy patients: a large questionnaire study.
Accumulating evidence suggests epilepsy and seizures may influence circadian rhythms and that circadian rhythms may influence epilepsy. It is also conceivable that seizure timing influences the timing of daily activities, sleeping, and wakefulness (i.e., chronotype). Only one group has studied the distribution of chronotypes of epileptics, showing significant differences between the diurnal activity patterns in two groups of patients with different epilepsy syndromes. The authors performed a questionnaire-based study of 200 epilepsy patients to compare the distribution of chronotypes and subjective sleep parameters of sleep duration and time of mid-sleep on free days to the distribution in the general population (n = 4042). Within this large group of epilepsy patients, we also compared the chronotypes of subsamples with well-defined epilepsy syndromes, i.e., temporal lobe epilepsy [TLE; n = 46], frontal lobe epilepsy [FLE; n = 30], and juvenile myoclonic epilepsy [JME; n = 38]. In addition, 27 patients who had had surgery for TLE were compared with those with TLE who had not had surgery. Both the Morningness-Eveningness Questionnaire and Munich Chronotype Questionnaire were used to determine chronotypes and subjective sleep parameters. Significant differences in morningness/eveningness distribution, timing of mid-sleep (corrected for sleep duration), and total sleep time on free days were found between epileptics and healthy controls. Those with epilepsy were more morning oriented, had earlier mid-sleep on free days, and longer sleep duration on free days (p < .001). However, distributions of chronotypes and sleep parameters between the groups of people with TLE, FLE, and JME did not differ. Persons who had surgery for TLE had similar morningness-eveningness parameters and similar sleep durations compared to those without surgery, but mid-sleep on free days was earlier in operated patients (p = .039). In conclusion, this is the first large study focusing on chronotypes in people with epilepsy. We show that the distribution of chronotypes and subjective sleep parameters of epileptics, in general, is different from that of healthy controls. Nevertheless, no differences are observed between patients with specified epilepsy syndromes, although they exhibit seizures with different diurnal patterns. Our results suggest that epilepsy, itself, rather than seizure timing, has a significant influence on chronotype behavior and subjective sleep parameters.


Loss of pVHL is sufficient to cause HIF dysregulation in primary cells but does not promote tumor growth.
Inactivation of the von Hippel-Lindau (VHL) gene is associated with the development of highly vascularized tumors. pVHL targets the alpha subunits of hypoxia inducible factor (HIF) for ubiquitin-mediated degradation in an oxygen-dependent manner. Although pVHL-deficient tumor cell lines demonstrate constitutive stabilization and activation of HIF, it has yet to be shown that loss of murine Vhl alone is sufficient to dysregulate HIF. We utilized a genetic approach to demonstrate that loss of Vhl is sufficient not only to stabilize HIF-alpha subunits under normoxia, but also fully activate HIF-mediated responses. These studies have implications for the hierarchy of signaling events leading to HIF stabilization, nuclear translocation, and target gene expression. We further demonstrate that loss of murine Vhl does not promote teratocarcinoma growth, indicating that other genetic changes must occur to facilitate Vhl-mediated tumorigenesis.


Physical and Mental Health Impacts of Household Gardens in an Urban Slum in Lima, Peru.
Rural poverty and lack of access to education has led to urban migration and fed the constant growth of urban slums in Lima, Peru. Inhabitants of these informal settlements lack land rights and access to a public water supply, resulting in poor sanitation, an inability to grow food, and suboptimal health outcomes. A repeated measures longitudinal pilot study utilizing participatory design methods was conducted in Lima between September 2013 and September 2014 to determine the feasibility of implementing household gardens and the subsequent impact of increased green space on well-being. Anthropometric data and a composite of five validated mental health surveys were collected at the baseline, 6-months, and 12-months after garden construction. Significant increases from the baseline in all domains of quality of life, including: physical (p < 0.01), psychological (p = 0.05), social (p = 0.02), environmental (p = 0.02), and overall social capital (p < 0.01) were identified 12 months after garden construction. Life-threatening experiences decreased significantly compared to the baseline (p = 0.02). There were no significant changes in parent or partner empathy (p = 0.21), BMI (p = 0.95), waist circumference (p = 0.18), or blood pressure (p = 0.66) at 6 or 12 months. Improved access to green space in the form of a household garden can significantly improve mental health in an urban slum setting.


Current smoking, former smoking, and adverse outcome among hospitalized COVID-19 patients: a systematic review and meta-analysis.
The purpose of this study was to examine the prevalence and effects of current smoking on adverse outcomes among hospitalized COVID-19 patients. A systematic review of the literature (PubMed) identified 18 (from a total of 1398) relevant studies. Pooled current smoking prevalence was compared with the gender-adjusted and gender and age-adjusted, population-based expected prevalence by calculating prevalence odds ratio (POR). The association between current, compared with non-current and former, smoking and adverse outcome was examined. A secondary analysis was performed by including 12 pre-publications (30 studies in total). All analyses were performed using random-effects meta-analysis. Among 6515 patients, the pooled prevalence of current smoking was 6.8% [95% confidence interval (CI): 4.8-9.1%]. The gender-adjusted POR was 0.20 (95% CI: 0.16-0.25, p < 0.001), and the gender and age-adjusted POR was 0.24 (95% CI: 0.19-0.30, p < 0.001). Current smokers were more likely to have an adverse outcome compared with non-current smokers [odds ratio (OR): 1.53, 95%CI: 1.06-2.20, p = 0.022] but less likely compared with former smokers (OR: 0.42, 95% CI: 0.27-0.74, p = 0.003). When pre-publications were added (n = 10,631), the gender-adjusted POR was 0.27 (95% CI: 0.19-0.38, p < 0.001) and the gender and age-adjusted POR was 0.34 (95% CI: 0.24-0.48, p < 0.001). This meta-analysis of retrospective observational case series found an unexpectedly low prevalence of current smoking among hospitalized patients with COVID-19. Hospitalized current smokers had higher odds compared with non-current smokers but lower odds compared with former smokers for an adverse outcome. Smoking cannot be considered a protective measure for COVID-19. However, the hypothesis that nicotine may have a protective effect in COVID-19 that is partially masked by smoking-related toxicity and by the abrupt cessation of nicotine intake when smokers are hospitalized should be explored in laboratory studies and clinical trials using pharmaceutical nicotine products.


[Minimally invasive emergent techniques in obesity treatment].
Obesity is a chronic disease for which several modalities of treatment are investigated today. One of them is the set of minimally aggressive techniques that have been added to the intragastric balloon. To review the minimally invasive techniques described in the last years for the treatment of obesity. It consisted in reviewing the bibliography through the habitual finders, in addition to the obtained data of the companies. They are classified in restrictive and malabsortive, and the restrictive are divides in mechanical or functional restriction. Between mechanical restrictive the classified as we included in the restrictive emergent techniques the adjustable intragastric balloon, the intragastric prosthesis, the vertical endoluminal gastroplasty and the transoral gastroplasty. In order to obtain a functional restriction, we have the gastric pacemaker and the botulinic toxin. And finally, the endoluminal duodenojejunal bypass is described as a malabsortive technique. With less than 10 years of existence, it seems that the described techniques compensate their smaller effectiveness compared to the surgical techniques, with the absence of substantial modifications in the anatomy of the alimentary tract. None of these techniques is free of risks and complications.


Meta-Analysis of Correlations Between Marginal Bone Resorption and High Insertion Torque of Dental Implants.
To evaluate correlations between marginal bone resorption and high insertion torque value (> 50 Ncm) of dental implants and to assess the significance of immediate and early/conventional loading of implants under a certain range torque value. Specific inclusion and exclusion criteria were used to retrieve eligible articles from Ovid, PubMed, and EBSCO up to December 2013. Screening of eligible studies, quality assessment, and data extraction were conducted in duplicate. The results were expressed as random/fixed-effects models using weighted mean differences for continuous outcomes with 95% confidence intervals. Initially, 154 articles were selected (11 from Ovid, 112 from PubMed, and 31 from EBSCO). After exclusion of duplicate articles and articles that did not meet the inclusion criteria, six clinical studies were selected. Assessment of P values revealed that correlations between marginal bone resorption and high insertion torque were not statistically significant and that there was no difference between immediately versus early/conventionally loaded implants under a certain range of torque. None of the meta-analyses revealed any statistically significant differences between high insertion torque and conventional insertion torque in terms of effects on marginal bone resorption.


National screening study on 10 perfluorinated compounds in raw and treated tap water in France.
The occurrence of seven perfluoroalkyl carboxylates (PFCAs) and three perfluoroalkyl sulfonates (PFASs) was studied in raw- and treated-water samples from public water systems. Two sampling campaigns were performed during the summer of 2009 and in June 2010. Sampling was equally distributed across the 100 French departments. In total, 331 raw-water samples and 110 treated-water samples were analyzed during this study, representing approximately 20% of the national water supply flow. Concentrations of perfluorinated compounds (PFCs) were determined using automated solid-phase extraction and liquid chromatography-tandem mass spectrometry. In raw-water samples, the highest individual PFC concentration was 139 ng/L for perfluorohexanoic acid (PFHxA). The sum of all of the determined components was >100 ng/L at three sampling points (199, 117, and 115 ng/L). Of the investigated PFCs, perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHxS), perfluorooctanoic acid (PFOA), and PFHxA predominated (detected in 27%, 13%, 11%, and 7% of samples, respectively). Geographical variability was observed, with departments crossed by major rivers or with high population densities being more affected by PFC contamination. Compared with raw water, short-chain PFCAs, but not PFASs, were found in higher abundance in treated water. This difference suggests a relative effectiveness of certain water treatments for the elimination of PFASs but also a possible degradation of PFCA precursors by water-treatment processes. Our investigations did not show any heavily contaminated sites. In treated-water samples, the highest individual PFC concentration was 125 ng/L for PFHxA. The sum of all of the determined components was >100 ng/L at one sampling point (156 ng/L). The values observed for PFOS and PFOA in drinking water were not greater than the health-based drinking-water concentration protectives for lifetime exposure that have been defined for other countries.


Phox2B correlates with MYCN and is a prognostic marker for neuroblastoma development.
Neuroblastoma is the one of the most common extracranial childhood malignancies, accounting for ∼15% of tumor-associated deaths in children. It is generally considered that neuroblastoma originates from neural crest cells in the paravertebral sympathetic ganglia and the adrenal medulla. However, the mechanism by which neuroblastoma arises during sympathetic neurogenesis and the cellular mechanism that drives neuroblastoma development remains unclear. The present study investigated the cell components during neuroblastoma development in the tyrosine hydroxylase-v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (TH-MYCN) mouse model, a transgenic mouse model of human neuroblastoma. The present study demonstrates that paired-like homeobox 2b (Phox2B)+ neuronal progenitors are the major cellular population in hyperplastic lesions and primary tumors. In addition, Phox2B+ neuronal progenitors in hyperplastic lesions or primary tumors were observed to be in an actively proliferative and undifferentiated state. The current study also demonstrated that high expression levels of Phox2B promotes neuroblastoma cell proliferation and xenograft tumor growth. These findings indicate that the proliferation of undifferentiated Phox2B+ neuronal progenitors is a cellular mechanism that promotes neuroblastoma development and indicates that Phox2B is a critical regulator in neuroblastoma pathogenesis.


The CD40-CD154 system in anti-infective host defense.
Research in the past few years has documented significant advances in our understanding of the CD40-CD40 ligand (CD154) system in diverse immune functions. This system influences many T cell mediated inflammatory immune responses and effector functions, unmasking a previously unexpected role for CD40-CD154 in cell mediated immunity. Manipulation of CD154 in animal models of infection by the use of CD154-deficient mice or anti-CD154 antibodies has shown the importance of this system in the initiation of the inflammatory response, in the activation of antigen-presenting cells and in resistance to infections.


Characterization and quantitation of three B-type lamins in Xenopus oocytes and eggs: increase of lamin LI protein synthesis during meiotic maturation.
We have previously shown that Xenopus oocytes, eggs, and early embryos contain lamins LII and LIII, and that portions of each are associated with distinct egg vesicle populations. We now report that a lamin similar or identical to the B-type lamin LI is also present in oocyte nuclei and in egg extracts. We have quantitated the three B-type lamins per oocyte nucleus, and have calculated relative ratios of LI:LIII = 1:100, and LII:LIII = 1:10. Similar to lamin LII, 5-15% of lamin LI is associated with egg membranes in a biochemically stable manner. Egg vesicles absorbed with lamin isoform-specific antibodies to magnetic beads indicate that lamin LI-associated egg membranes are of heterogenous morphology, and are independent from the lamin LII and LIII vesicle populations. Compared to other nuclear envelope proteins, the synthesis of lamin LI protein is specifically elevated during meiotic maturation, resulting in a 4- to 12-fold higher amount of lamin LI in eggs than is present in oocyte nuclei. Immunofluorescence and immunoblot analysis demonstrated that lamins LI, LII, and LIII are associated with the nuclear envelope formed on demembranated sperm when added to activated egg extract. These results strongly suggest that three different lamin-associated vesicle populations are involved in the formation of a nuclear envelope in egg extracts.


Implementation planning and progress on physical activity goals: the mediating role of life-management strategies.
This 4-week prospective study examined whether the use of life-management strategies mediates the relationship between implementation planning and short-term progress on physical activity goals. In particular the strategies of elective selection, compensation, and loss-based selection were disentangled to assess their specific mediating effects. Results from a sample of 131undergraduate students showed that, as a composite, life-management strategies fully mediated the relationship between planning and goal progress. More specifically, decomposing the effects demonstrated that only elective selection and compensation mediated the association between planning and greater progress on a personal physical activity goal. Results are discussed in light of their practical implications and contributions to the personal goal literature.


Effect of contrast, stimulus density, and viewing distance on multifocal steady-state visual evoked potentials (MSVs).
We investigated the effects of image contrast, stimulus density, and viewing distance upon a multifocal steady-state visual evoked potential (MSV) method. Fourteen adults with normal vision (mean age = 27.0 ± 6.6 years; 6 males) participated in the study. Each of the stimulus regions of the multifocal ensembles presented a contrast modulated grating, displaying spatial and temporal frequencies that evoke the spatial frequency doubling illusion. All subjects were tested at five contrasts: 0.06, 0.11, 0.22, 0.45, and 0.89; viewed at 16, 32, and 128 cm. A multivariate linear model estimated factors for each stimulus region, recording channel, number of stimuli (9 or 17 regions), and sex; and covariates for contrast, and octaves of viewing distance, and age. The responses per unit area for the 17-region display were significantly larger than for the 9-region display (P < 10⁻¹²). The contrast-response function could be described by a power law with exponent 0.068 (P < 0.008). The effect of viewing distance was small but significant: response amplitude dropped by -0.17 ± 0.03 dB per octave of viewing distance (P < 10⁻⁶), or 10% over 8 octaves. The response per unit area indicated that cortical folding diminishes responses to larger stimuli. Viewing distance did not greatly affect response amplitude. This suggested that we can use similar, but scaled, stimuli to study central and peripheral disease. The rapidly saturating contrast responses imply that there would be nothing lost from testing at contrasts as low as 20% given that higher, saturating contrasts might mask visual field defects.


Sneakers and spokes: an assessment of the walkability and bikeability of U.S. postsecondary institutions.
The purpose of the study described in this article was to assess the walkability and bikeability of 15 U.S. postsecondary education campuses. The Centers for Disease Control and Prevention's evidence-based Healthier Worksite Initiative Walkability Audit was modified to rate campus walking and biking path segments for path safety, quality, and comfort. Universities (n = 13) assessed an average of 44 path segments, which earned a mean score of 72.71 +/- 10.77 SD (possible range 0 to 100). Postsecondary technical schools (n = 2) assessed 20 path segments, which received an average score of 76.56 +/- 13.15. About 70% of path segments received a grade A or B, but almost 1 in 10 received a failing or poor support score for walking and biking. Nearly half or more campus environments scored significantly below an acceptable score on many path safety and quality criteria. Postsecondary education campuses that are supportive of walking and biking offer numerous benefits to the environment and people. Findings from environmental assessments like the data reported here can provide valuable input to campus planners.


Degenerative disease in rotator cuff tears: what are the biochemical and histological changes?
The histopathological changes associated with rotator cuff tears include thinning and disorganization of collagen fibers, the presence of granulation tissue, increased levels of glycosaminoglycans, fibrocartilaginous metaplasia, calcification, fatty infiltration, and necrosis of the tendon margin with cell apoptosis. The biochemical changes include an increase in the expression of matrix metalloproteinases (MMPs) and a decrease in tissue inhibitor of metalloproteinases (TIMP) messenger ribonucleic acid expression. Histological evidence of tendinopathy has been found in patients with rotator cuff tear. Biochemical changes include significant increases in MMP1, MMP2, MMP3, and in TIMP1 and TIMP2 levels, not only at the lateral supraspinatus edge, but also in the macroscopically intact portion of the supraspinatus tendon and in the intact subscapularis. The tissue in the ruptured area of the supraspinatus tendon undergoes marked rearrangement at molecular levels. This involves the activity of MMP1, 2, and 3 and supports a critical role of MMPs in tendon physiology. Intact parts of the torn supraspinatus tendon can present the histopathological changes associated with rotator cuff tears. These findings suggest that biochemical changes can already occur in a macroscopically intact tendon and seem to point to a global degenerative process in the shoulder.


Contrasted coevolutionary dynamics between a bacterial pathogen and its bacteriophages.
Many antagonistic interactions between hosts and their parasites result in coevolution. Although coevolution can drive diversity and specificity within species, it is not known whether coevolutionary dynamics differ among functionally similar species. We present evidence of coevolution within simple communities of Pseudomonas aeruginosa PAO1 and a panel of bacteriophages. Pathogen identity affected coevolutionary dynamics. For five of six phages tested, time-shift assays revealed temporal peaks in bacterial resistance and phage infectivity, consistent with frequency-dependent selection (Red Queen dynamics). Two of the six phages also imposed additional directional selection, resulting in strongly increased resistance ranges over the entire length of the experiment (ca. 60 generations). Cross-resistance to these two phages was very high, independent of the coevolutionary history of the bacteria. We suggest that coevolutionary dynamics are associated with the nature of the receptor used by the phage for infection. Our results shed light on the coevolutionary process in simple communities and have practical application in the control of bacterial pathogens through the evolutionary training of phages, increasing their virulence and efficacy as therapeutics or disinfectants.


From Mating to Milk Access: A Review of Reproductive Vocal Communication in Mice.
Vocalisations play a central role in rodent communication, especially in reproduction related behaviours. In adult mice (Mus musculus) the emission of ultrasonic vocalisations (USVs) has been observed in courtship and mating behaviour, especially by males. These have been found to have distinctive individual signatures that influence female choice of mating partner. The most recent findings show that vocal communication also has a role in parental cooperation, in that female mice communicate with male partners in ultrasonic frequencies to induce paternal behaviour. Infant vocalisations form the other important part of reproductive vocal communication. Although born deaf, neonatal mice are capable of producing vocalisations since birth. As an altricial species, successful mother-infant communication is essential for survival, and these vocalisations are important modulators of maternal behaviour. Three main types of infant vocalisations have been identified and characterised. Most research has addressed pure USVs, related to stressful situations (e.g., cold, isolation, handling, presence of unfamiliar males or predators), which usually elicit maternal search and retrieval. In addition, broad-band spectrum signals, emitted post-partum during cleaning of foetal membranes, inhibit biting and injury by adults and "wriggling calls," emitted during suckling, release maternal behaviour (such as licking). Several variables have been identified to modulate vocalisations in mice, including individual characteristics such as strain/genotype, age, sex, and experimental factors such as pharmacological compounds and social context. In recent years, there has been a big increase in the knowledge about the characteristics of vocal communication in rodents due to recent technological advances as well as a growing interest from the neuroscience community. Vocalisation analysis has become an essential tool for phenotyping and evaluating emotional states. In this review, we will (i) provide a comprehensive summary of the current knowledge on mouse reproductive vocal communication and (ii) discuss the most recent findings in order to provide a broad overview on this topic.


Sexism-Related Stigma Affects Pain Perception.
People with stigmatized characteristics tend to be devalued by others in a given society. The negative experiences related to stigma cause individuals to struggle as they would if they were in physical pain and bring various negative outcomes in the way that physical pain does. However, it is unclear whether stigma related to one's identity would affect their perception of physical pain. To address this issue, using sexism-related paradigms, we found that females had reduced pain threshold/tolerance in the Cold Pressor Test (Experiment 1) and an increased rating for nociceptive laser stimuli with fixed intensity (Experiment 2). Additionally, we observed that there was a larger laser-evoked N1, an early laser-evoked P2, and a larger magnitude of low-frequency component in laser-evoked potentials (LEPs) in the stigma condition than in the control condition (Experiment 3). Our study provides behavioral and electrophysiological evidence that sexism-related stigma affects the pain perception of females.


Coming down from the trees: is terrestrial activity in Bornean orangutans natural or disturbance driven?
The orangutan is the world's largest arboreal mammal, and images of the red ape moving through the tropical forest canopy symbolise its typical arboreal behaviour. Records of terrestrial behaviour are scarce and often associated with habitat disturbance. We conducted a large-scale species-level analysis of ground-based camera-trapping data to evaluate the extent to which Bornean orangutans Pongo pygmaeus come down from the trees to travel terrestrially, and whether they are indeed forced to the ground primarily by anthropogenic forest disturbances. Although the degree of forest disturbance and canopy gap size influenced terrestriality, orangutans were recorded on the ground as frequently in heavily degraded habitats as in primary forests. Furthermore, all age-sex classes were recorded on the ground (flanged males more often). This suggests that terrestrial locomotion is part of the Bornean orangutan's natural behavioural repertoire to a much greater extent than previously thought, and is only modified by habitat disturbance. The capacity of orangutans to come down from the trees may increase their ability to cope with at least smaller-scale forest fragmentation, and to cross moderately open spaces in mosaic landscapes, although the extent of this versatility remains to be investigated.


Social interactions in virtual reality exposure therapy: A proof-of-concept pilot study.
Research on virtual reality exposure therapy (VRET) has demonstrated good treatment efficacy with regards to several anxiety disorders. Yet, there is lack of knowledge about the value of integrating interaction between clients and virtual humans in VRET. Such interaction might prove effective in treating psychological complaints that involve social interactions, such as social anxiety. A VRET system specifically designed to expose clients with social anxiety disorder to anxiety provoking social situations was applied to 16 and 18 individuals with high and low levels of social anxiety, respectively. Participants engaged in two exposure sessions in several free speech dialogues with virtual humans while being monitored by a therapist. Participants with high levels of social anxiety reported significantly lower levels of social anxiety three months after exposure to two virtual reality interaction sessions than before treatment (p < 0.01). In the group with low levels of social anxiety, no significant change of social anxiety was reported between pre-treatment and follow-up. Additionally, participants in both groups reported higher self-efficacy three months after treatment than before treatment (ps ≤ 0.001). These findings indicate that virtual reality technology that incorporates social interactions may be successfully applied for therapeutic purposes.


Insulin-like growth factor 1 attenuates antiestrogen- and antiprogestin-induced apoptosis in ER+ breast cancer cells by MEK1 regulation of the BH3-only pro-apoptotic protein Bim.
In this pre-clinical in vitro study conducted in estrogen receptor positive (ER+) breast cancer cells, we have characterized the effects of insulin-like growth factor I (IGF-1) on the cytostatic and cytotoxic action of antiestrogen treatment when used as a single agent or in combination with the antiprogestin mifepristone (MIF). Our goal was to identify new molecular targets to improve the efficacy of hormonal therapy in breast cancer patients that have a poor response to hormonal therapy, in part, due to high circulating levels of unbound insulinIGF-1. IGF-1-mediated effects on cytostasis and apoptotic cell death were determined with cell counts conducted in the presence and absence of trypan blue; enzyme-linked immunosorbent assays to determine the intracellular levels of cleaved cytokeratin 18, a marker of epithelial cancer cell apoptosis; and immunoblot analysis to determine the levels of cleaved poly-ADP ribose polymerase (PARP) and lamin A that result from caspase-dependent apoptosis. Cytotoxicity was further characterized by determination of the levels of reactive oxygen species (ROS) and the percent of mitochondrial membrane depolarization in cell populations treated with the different hormones in the presence and absence of IGF-1. Small molecule inhibitors of the dual-specificity protein kinase MEK1, MEK1 siRNA, Bim siRNA, and vectors overexpressing MEK1 wild type and mutant, dominant negative cDNA were used to identify key IGF-1 downstream prosurvival effectors. IGF-1, at physiologically relevant levels, blocked the cytotoxic action(s) of the antiestrogens 4-hydroxytamoxifen (4-OHT) and tamoxifen (TAM) when used as single agents or in combination with the antiprogestin MIF. The antiapoptotic action of IGF-1 was mediated primarily through the action of MEK1. MEK1 expression reduced the levels of ROS and mitochondrial membrane depolarization induced by the hormonal treatments via a mechanism that involved the phosphorylation and proteasomal turnover of the proapoptotic BH3-only Bcl-2 family member Bim. Importantly, small-molecule inhibitors of MEK1 circumvented the prosurvival action of IGF-1 by restoring Bim to levels that more effectively mediated apoptosis in ER+ breast cancer cells. his study provides strong support for the use of MEK1 inhibitors in combination with hormonal therapy to effectively affect cytostasis and activate a Bim-dependent apoptotic pathway in ER+ breast cancer cells. We discuss that MEK1 blockade may be a particularly effective treatment for women with high circulating levels of IGF-1, which have been correlated to a poor prognosis.


Mammary Gland Involution Provides a Unique Model to Study the TGF-β Cancer Paradox.
Transforming Growth Factor-β (TGF-β) signaling in cancer has been termed the "TGF-β paradox", acting as both a tumor suppresser and promoter. The complexity of TGF-β signaling within the tumor is context dependent, and greatly impacted by cellular crosstalk between TGF-β responsive cells in the microenvironment including adjacent epithelial, endothelial, mesenchymal, and hematopoietic cells. Here we utilize normal, weaning-induced mammary gland involution as a tissue microenvironment model to study the complexity of TGF-β function. This article reviews facets of mammary gland involution that are TGF-β regulated, namely mammary epithelial cell death, immune activation, and extracellular matrix remodeling. We outline how distinct cellular responses and crosstalk between cell types during physiologically normal mammary gland involution contribute to simultaneous tumor suppressive and promotional microenvironments. We also highlight alternatives to direct TGF-β blocking anti-cancer therapies with an emphasis on eliciting concerted microenvironmental-mediated tumor suppression.


Connected, attracted, and concerned: A Q study on medical crossborder curriculum partnerships.
A new form of internationalization has been trending upward in the medical education realm: crossborder medical curriculum partnerships established to deliver the same, or adapted, curriculum to groups of geographically separated students. This study aims to investigate crossborder medical curriculum partnerships by exploring the experiences of teachers at the recipient institution who have a key role in delivering the program. From four pioneering recipient medical schools, 24 teachers participated in a Q-sort study. Each participant rank-ordered 42 statements about teaching in a crossborder medical curriculum on a scale from -5 (indicating strong disagreement) to +5 (indicating strong agreement). The authors conducted a "by-person" factor analysis to uncover distinct patterns in the ranking of statements, using the statistical results and participants' comments about their Q sorts to interpret these patterns and translate them into distinct viewpoints. Three viewpoints emerged, reflecting: (1) a feeling of connectedness with the partner institution, trust in the quality of the curriculum, and appreciation of interinstitutional relationships; (2) the partnership's attractiveness because of the career opportunities it offers; and (3) concerns over the quality of graduates because of doubts about the appropriateness of the didactic model and insufficient attention to local healthcare needs, and over the practical feasibility of such partnerships. The three viewpoints identified revealed a pallet of views on how host teachers might experience their work. It shows the heterogeneous features of this group and seems to counterbalance reports that they are feeling "deprived" from their role as teacher. Two viewpoints featured an appreciation of interinstitutional relationships and of the partnership, especially when perceiving a degree of autonomy. Partners can capitalize on all different viewpoints by deploying procedure and policies to raise the quality of education delivery.


Impact of mammography on U.S. breast cancer mortality, 1975-2000: are intermediate outcome measures informative?
Seven models have estimated the contribution of screening to the decrease in U.S. breast cancer mortality between 1975 and 2000. We will investigate whether the model estimates of the mortality reduction due to screening are associated with intermediate outcome measures (IOMs). Detection rates at screening, 1- and 2-year sensitivity, program sensitivity, and incidence of advanced tumors are used as IOMs. Moreover, the model parameters preclinical duration and sensitivity are analyzed. The correlation of IOMs with mortality is assessed for actual U.S. screening and for an intensive screening scenario, with annual screening at ages 40-79 years with 100% participation. Also, 12 alternative screening scenarios are run for one of the models, and within-model correlation between IOMs and mortality reduction is described. Resulting correlations between IOMs and mortality reduction are mostly weak. For 2-year sensitivity and the incidence of advanced tumors, correlations are high in the intensive screening scenario. Within-model correlations are strong for incidence of advanced tumors and program sensitivity. Intermediate outcome measures have limited potential in predicting the impact of mammographic screening on mortality. Incidence of advanced tumors and program sensitivity are measures that merit further consideration as surrogates for mortality reduction.


Current comprehensive management of cranial base chordomas: 10-year meta-analysis of observational studies.
The role of surgery and adjuvant radiation therapy for cranial base chordomas is not well established. This meta-analysis measures the relationship of complete resection and type of adjuvant radiation therapy to 5-year progression-free survival (PFS) and overall survival (OS) of cranial base chordomas. A systematic MEDLINE search (1999-present) yielded 23 observational studies and 807 patients who fit inclusion criteria. The following analyses were performed: 1) Kaplan-Meier 5-year PFS and OS compared based on the extent of resection and type of adjuvant radiation therapy using the log-rank method; 2) a random-effects model comparing 5-year PFS with complete or incomplete resection; and 3) paired z-test comparisons of weighted average 5-year OS and PFS grouped by type of adjuvant radiation therapy. The weighted average follow-up was 53.6 months. The weighted average 5-year PFS and OS were 50.8% and 78.4%, respectively. Complete resection conferred a higher 5-year PFS than incomplete resection from the random effects model (mean difference in PFS 20.7%; 95% CI 6.57%-34.91%). Patients with incomplete resection were 3.83 times more likely to experience a recurrence (95% CI 1.63-9.00) and 5.85 times more likely to die (95% CI 1.40-24.5) at 5 years versus patients with complete resection. There was no difference in 5-year OS by type of adjuvant radiation, although 5-year PFS was lower in patients receiving Gamma Knife surgery relative to carbon ion radiotherapy (p = 0.042) on paired z-test. No survival difference occurred between radiation therapy techniques on Kaplan-Meier analysis of compiled patient data. Patients with complete resection of cranial base chordomas have a prolonged 5-year PFS and OS. Adjuvant proton-beam, carbon ion, and modern fractionated photon radiation therapy techniques offered a similar rate of PFS and OS at 5 years.


Development of non-antibiotic-resistant, chromosomally based, constitutive and inducible expression systems for aroA-attenuated Salmonella enterica Serovar Typhimurium.
Live-vaccine delivery systems expressing two model antigens from Mycoplasma hyopneumoniae, F2(P97) (Adh) and NrdF, were constructed using Salmonella enterica serovar Typhimurium aroA (STM-1), and immunogenicity in mice was evaluated. Recombinant plasmid-based expression (PBE) and chromosomally based expression (CBE) systems were constructed. The PBE system was formed by cloning both antigen genes into pJLA507 to create an operon downstream of temperature-inducible promoters. Constitutive CBE was achieved using a promoter-trapping technique whereby the promoterless operon was stably integrated into the chromosome of STM-1, and the expression of antigens was assessed. The chromosomal position of the operon was mapped in four clones. Inducible CBE was obtained by using the in vivo-induced sspA promoter and recombining the expression construct into aroD. Dual expression of the antigens was detected in all systems, with PBE producing much larger quantities of both antigens. The stability of antigen expression after in vivo passage was 100% for all CBE strains recovered. PBE and CBE strains were selected for comparison in a vaccination trial. The vaccine strains were delivered orally into mice, and significant systemic immunoglobulin M (IgM) and IgG responses against both antigens were detected among all CBE groups. No significant immune response was detected using PBE strains. Expression of recombinant antigens in S. enterica serovar Typhimurium aroA from chromosomally located strong promoters without the use of antibiotic resistance markers is a reliable and effective method of inducing a significant immune response.


Influence of humidity on tribo-electric charging and segregation in shaken granular media.
We study the effect of humidity on the charge accumulation of polymer granulates shaken vertically in a stainless steel container. This setup allows us to control the humidity level from 5% to 100%RH while performing automated charge measurements in a Faraday cup directly connected to the shaking container. We find that samples of approximately 2000 polymer spheres become highly charged at low humidity levels (<30%RH), but acquire almost no charge for humidity levels above 80%RH. The transition between these two regimes does depend on the material, as does the sign of the charge. For the latter we find a correlation with the contact angle of the polymer with only very hydrophilic particles attaining positive charges. We show that this humidity dependence of tribo-charging can be used to control segregation in shaken binary mixtures.


Human IgG Fc domain engineering enhances antitoxin neutralizing antibody activity.
The effector activity of antibodies is dependent on engagement with Fcγ receptors (FcγRs) and activation of the associated intracellular signaling pathways. Preclinical evaluation of therapeutic humanized or chimeric mAbs to study the interactions of their Fc regions with FcγRs is hampered by substantial structural and functional FcγR diversity among species. In this report, we used mice expressing only human FcγRs to evaluate the contribution of FcγR-mediated pathways to the neutralizing activity of an anti-anthrax toxin chimeric mAb. We observed that the protective activity of this mAb was highly dependent upon FcγR engagement, with minimal protection against anthrax toxin observed in FcγR-deficient mice following mAb administration. We generated anti-anthrax toxin mAbs with specific Fc domain variants with selectively enhanced affinity for particular human FcγRs and assessed their activity in FcγR-humanized mice. We determined that Fc domain variants that were capable of selectively engaging activating FcγRs substantially enhanced the in vitro and in vivo activity of anthrax toxin-neutralizing antibodies. These findings indicate that the application of Fc domain engineering is a feasible strategy to enhance toxin-neutralizing activity and suggest that engineered antitoxin antibodies will have improved therapeutic efficacy.


Adult polyglucosan body disease associated with lewy bodies and tremor.
Adult polyglucosan body disease (PGBD) is rare and typically presents with upper and lower motor neuron involvement and neurogenic bladder. Extrapyramidal features are unusual in PGBD and are presumed secondary to widespread pathology that includes the basal ganglia. There are no prior reports of Lewy bodies in PGBD. To report a unique finding of Lewy bodies in a patient with PGBD. REPORT OF A CASE A 46-year-old woman initially presented with a 4-year history of resting tremor. The tremor responded to levodopa therapy. Several months later, she developed upper and lower motor neuron involvement and other clinical features of PGBD. A sural nerve biopsy specimen revealed intra-axonal polyglucosan bodies that confirmed the clinical diagnosis. Bulbar and limb weakness progressed, and she developed dementia. She died 6 years after onset. At autopsy, extensive polyglucosan body formation was found in many regions of the central nervous system. In addition, numerous alpha-synuclein staining Lewy bodies were observed in the substantia nigra, accompanied by marked neuron depopulation. To our knowledge, this is the first report of adult PGBD associated with Lewy bodies and levodopa-responsive tremor. Although polyglucosan bodies were seen in substantia nigra, it is most likely that our patient had coexisting Parkinson disease.


The selective peroxisome proliferator-activated receptor alpha modulator (SPPARMα) paradigm: conceptual framework and therapeutic potential : A consensus statement from the International Atherosclerosis Society (IAS) and the Residual Risk Reduction Initiative (R3i) Foundation.
In the era of precision medicine, treatments that target specific modifiable characteristics of high-risk patients have the potential to lower further the residual risk of atherosclerotic cardiovascular events. Correction of atherogenic dyslipidemia, however, remains a major unmet clinical need. Elevated plasma triglycerides, with or without low levels of high-density lipoprotein cholesterol (HDL-C), offer a key modifiable component of this common dyslipidemia, especially in insulin resistant conditions such as type 2 diabetes mellitus. The development of selective peroxisome proliferator-activated receptor alpha modulators (SPPARMα) offers an approach to address this treatment gap. This Joint Consensus Panel appraised evidence for the first SPPARMα agonist and concluded that this agent represents a novel therapeutic class, distinct from fibrates, based on pharmacological activity, and, importantly, a safe hepatic and renal profile. The ongoing PROMINENT cardiovascular outcomes trial is testing in 10,000 patients with type 2 diabetes mellitus, elevated triglycerides, and low levels of HDL-C whether treatment with this SPPARMα agonist safely reduces residual cardiovascular risk.


Alsin is partially associated with centrosome in human cells.
Mutations in the ALS2 gene has recently been linked to cases of juvenile amyotrophic lateral sclerosis, juvenile primary lateral sclerosis and ascending hereditary spastic paralysis. All reported mutations predict the production of truncated forms of Alsin suggesting a loss of function mechanism for these motor neuron disorders. Here we used the tetracycline-regulated expression system to overexpress the full-length and truncated forms of Alsin in different cell lines. Alsin overexpression caused severe phenotypic changes in monkey COS-7 cells including the enlargement and accumulation of early endosomes, impairment of mitochondria trafficking and fragmentation of the Golgi apparatus. Our results further demonstrate the requirement of the Alsin VPS9 domain for occurrence of the vacuolation process and the role of Alsin as a guanine nucleotide exchange factor for Rab5. Transfected human SW13 cells exhibited an unexpected centrosomal localization for Alsin that was linked to the presence of the c-terminal part of the protein. Immunofluorescence staining revealed a colocalization of Alsin with the centrosomal markers gamma-tubulin and A kinase anchoring protein (AKAP-450). Similar results were obtained with human LA-N-2 and SK-N-SH neuronal cells. Moreover endogenous Alsin was detected in a centrosome preparation purified from human cortical brain. Considering the crucial role of centrosome in the production of microtubules required for intracellular transport, these findings are of potential relevance for unravelling the disease mechanisms linked to Alsin mutations.


Project GRACE: a staged approach to development of a community-academic partnership to address HIV in rural African American communities.
The HIV epidemic is a health crisis in rural African American communities in the Southeast United States; however, to date little attention has been paid to community-academic collaborations to address HIV in these communities. Interventions that use a community-based participatory research (CBPR) approach to address individual, social, and physical environmental factors have great potential for improving community health. Project GRACE (Growing, Reaching, Advocating for Change and Empowerment) uses a CBPR approach to develop culturally sensitive, feasible, and sustainable interventions to prevent the spread of HIV in rural African American communities. This article describes a staged approach to community-academic partnership: initial mobilization, establishment of organizational structure, capacity building for action, and planning for action. Strategies for engaging rural community members at each stage are discussed; challenges faced and lessons learned are also described. Careful attention to partnership development has resulted in a collaborative approach that has mutually benefited both the academic and community partners.


Gain-of-function mutations in TRPV4 cause autosomal dominant brachyolmia.
The brachyolmias constitute a clinically and genetically heterogeneous group of skeletal dysplasias characterized by a short trunk, scoliosis and mild short stature. Here, we identify a locus for an autosomal dominant form of brachyolmia on chromosome 12q24.1-12q24.2. Among the genes in the genetic interval, we selected TRPV4, which encodes a calcium permeable cation channel of the transient receptor potential (TRP) vanilloid family, as a candidate gene because of its cartilage-selective gene expression pattern. In two families with the phenotype, we identified point mutations in TRPV4 that encoded R616Q and V620I substitutions, respectively. Patch clamp studies of transfected HEK cells showed that both mutations resulted in a dramatic gain of function characterized by increased constitutive activity and elevated channel activation by either mechano-stimulation or agonist stimulation by arachidonic acid or the TRPV4-specific agonist 4alpha-phorbol 12,13-didecanoate (4alphaPDD). This study thus defines a previously unknown mechanism, activation of a calcium-permeable TRP ion channel, in skeletal dysplasia pathogenesis.


"Deafness" effects in detecting alterations to auditory feedback during sequence production.
Past research has shown that when discrete responses are associated with a perceptual goal, performers may have difficulty detecting stimuli that are commensurate with that goal. Three experiments are reported here that test whether such effects extend to sequence production. In Experiment 1, participants performed 8-note melodies repeatedly, and on each trial a single tone could be altered with respect to its pitch and/or synchrony with actions. Results suggested a selective deficit of detection when feedback pitch was unchanged and the event was slightly delayed. Experiment 2 showed that this "deafness" to feedback is limited to rhythmic motor tasks that require sequencing, in that similar effects did not emerge when participants produced pitch sequences by tapping a single key repeatedly. A third experiment demonstrated similar results to Experiment 1 when the mapping of keys to pitches on the keyboard was reversed. Taken together, results suggest a selective deafness to response-congruent delayed feedback, consistent with the idea that performers suppress previously planned events during production.


A review of basic to clinical studies of irreversible electroporation therapy.
The use of irreversible electroporation (IRE) for cancer treatment has increased sharply over the past decade. As a nonthermal therapy, IRE offers several potential benefits over other focal therapies, which include 1) short treatment delivery time, 2) reduced collateral thermal injury, and 3) the ability to treat tumors adjacent to major blood vessels. These advantages have stimulated widespread interest in basic through clinical studies of IRE. For instance, many in vitro and in vivo studies now identify treatment planning protocols (IRE threshold, pulse parameters, etc.), electrode delivery (electrode design, placement, intraoperative imaging methods, etc.), injury evaluation (methods and timing), and treatment efficacy in different cancer models. Therefore, this study reviews the in vitro, translational, and clinical studies of IRE cancer therapy based on major experimental studies particularly within the past decade. Further, this study provides organized data and facts to assist further research, optimization, and clinical applications of IRE.


Non-agricultural work injuries among youth: a systematic review.
To systematically review the quantitative literature on factors associated with youth non-agricultural work injury. Seven electronic databases were searched for studies published between 1980 and 2005. In addition, reference lists from each potentially eligible study were checked and experts in the field contacted for additional studies. Studies had to meet relevance and quality appraisal criteria. Nine cross-sectional studies using multivariate analyses met the inclusion and quality appraisal criteria. This best evidence synthesis found that work injury varied significantly with job and workplace factors such as hazard exposure and perceived work overload. Visible minority status was also associated with likelihood of a work injury. The lack of youth work injury studies assessing the following factors was also identified: physical and cognitive development, safety training, supervision, social environment of the workplace, and intervention studies. This review has potential implications for prevention of work injuries. First, interventions need to target modifiable risk factors. This systematic review pointed to two job/workplace factors that are potentially modifiable: hazard exposure and work pace pressure. Second, the multiple determinants of work injury highlight the need to develop interventions and policies that focus on multiple factors rather than one-dimensional approaches that target a specific factor.


Treatment patterns of multiple sclerosis patients: a comparison of veterans and non-veterans using the NARCOMS registry.
Multiple sclerosis (MS) is a chronic illness of the central nervous system, with a highly variable clinical course. Available therapies are only partially effective and as a consequence treatment patterns between patients can be varied. Longitudinal databases consisting of large cohorts where successive and sequential data is collected may reveal disease and treatment characteristics not apparent when data is gathered during clinical trials that consist usually of relatively homogeneous patients followed for short durations. We analysed data from the North American Research Committee on Multiple Sclerosis registry, a self-reported database, to assess MS patient characteristics and treatment patterns, with a focus on veterans. We show that the Veteran Healthcare Administration (VHA) system of medical centres care for a greater number of patients with higher average disability but not necessarily patients who report primary progressive or actively worsening disease. We also show that the VHA medical centres appear to better provide multidisciplinary care, particularly in the areas of social work, physical therapy and urology. In general, treatment patterns for symptomatic therapies follow similar patterns across veterans and non-veterans groups. Treatment patterns for immunomodulatory agents suggest that VHA veterans use IMA less frequently than either non-VHA veterans or non-veterans.


Selective processing of social threat cues following acute tryptophan depletion.
The aim of the present study was to investigate the effects of low dose tryptophan depletion on recovered depressed patients both on and off antidepressant medication on an emotional Stroop task using social threat cues.Twenty-four healthy volunteers, 24 euthymic volunteers with a history of depression not currently on antidepressant medication and 24 euthymic volunteers with a history of depression and currently on antidepressant medication were randomly allocated to double-blind treatment with either a tryptophan depleting or a balanced mixture. All participants then completed subjective mood ratings and an emotional Stroop task using social threat cues. The recovered depressed group on medication demonstrated an increase in selective processing of social threat cues on the emotional Stroop task in the tryptophan depletion compared to the control condition. This was not the case for either healthy controls or the recovered depressed group not on medication. Although none of the patients showed a clinically significant relapse following tryptophan depletion, the medicated group showed a small but statistically significant increase in self-rated depression on the Profile of Mood States (POMS). Our data indicate that low-dose acute tryptophan depletion elicits both cognitive processing typical of the depressed state and subtle changes in subjective mood in the recovered depressed group on medication, but not in the recovered depressed group not on medication. This suggests that these two groups may differ in their underlying vulnerability to compromised serotonin function.


The extent of diffusion MRI markers of neuroinflammation and white matter deterioration in chronic schizophrenia.
In a previous study we have demonstrated, using a novel diffusion MRI analysis called free-water imaging, that the early stages of schizophrenia are more likely associated with a neuroinflammatory response and less so with a white matter deterioration or a demyelination process. What is not known is how neuroinflammation and white matter deterioration change along the progression of the disorder. In this study we apply the free-water measures on a population of 29 chronic schizophrenia subjects and compare them with 25 matching controls. Our aim was to compare the extent of free-water imaging abnormalities in chronic subjects with the ones previously obtained for subjects at their first psychotic episode. We find that chronic subjects showed a limited extent of abnormal increase in the volume of the extracellular space, suggesting a less extensive neuroinflammatory response relative to patients at the onset of schizophrenia. At the same time, the chronic schizophrenia subjects had greater extent of reduced fractional anisotropy compared to the previous study, suggesting increased white matter deterioration along the progression of the disease. Our findings substantiate the role of neuroinflammation in the earlier stages of the disorder, and the effect of neurodegeneration that is worsening in the chronic phase.


[A method for evaluating visual impairments using operant behavior in mice].
A behavioral method, using a shuttle box (one of the conditioned behaviors) for assessing visual function in mice, was investigated. Normal ICR mice were trained to avoid the unconditioned stimulus (electric footshock) during the presentation of conditioned stimuli (tone and light). After acquisition of avoidance response, normal ICR mice were presented randomly tone or light as the conditioned stimulus. The percent of avoidance in the presentation of light was decreased suddenly, but that of tone was not. However, the decrease of avoidance response to light stimulus was recovered by following trainings. The mice, which had acquired the condition response to tone and light stimulus alone, were treated in their cornea with 4N NaOH. The decrease of percent of avoidance in the mice with alkali-treated cornea after differentiation of conditioned stimuli did not recover following training. These results suggested that visual impairment of mice can be detected by this method. Therefore, we attempted to detect the visual impairments of dominant (Cts) and recessive (cac) hereditary cataract mice, hereditary retina degenerated mice (C3H) and first filial generation mice of C3H, F1 [(ICR X C3H) F1]. The acquisition curves of both Cts and cac cataract mice were similar to that of normal ICR mice, but those of C3H and F1 were similar to that of ICR mice treated with alkali. Moreover, changes in recovery %, which was calculated from avoidance response in the presentation of light alone after differentiation of conditioned stimuli, corresponded to the above results. These findings demonstrated that the visual impairment of Cts and cac cataract mice is weak, while that of C3H and F1 mice is strong.


Lack of Seed Coat Contamination with Cucumber mosaic virus in Lupin Permits Reliable, Large-Scale Detection of Seed Transmission in Seed Samples.
Sowing seed stocks with minimal virus content provides a key control measure in preventing damaging epidemics of Cucumber mosaic virus (CMV) in crops of narrow-leafed lupin (Lupinus angustifolius). A seed testing service provides an estimate of percent CMV infection based on a dry seed test in which bulked subsamples of ungerminated seed are ground to a fine powder for testing. When enzyme-linked immunosorbent assay (ELISA) was used, CMV antiserum that gave low background optical density (A405) values with extracts of powder from subsamples of healthy seed provided greatest accuracy, readily detecting one infected seed in subsamples of 100 seeds. In comparative ELISAs on duplicate subsamples from eight different seed stocks, germination and dry seed tests always gave similar percent infection values. When seed coats were separated from the embryos of CMV-infected and healthy lupin seeds before testing by ELISA, the virus was only detected in embryos from infected seeds and never in their seed coats. Treatment with trisodium phosphate did not alter the low ELISA optical density (A405) values obtained with seed coats separated from infected seeds. Therefore, seed coat contamination with CMV is lacking in lupin, justifying large-scale routine use of a dry seed test to estimate percent virus infection in commercial seed samples.


Health promotion when the 'vaccine' does not work.
The epidemics of obesity, metabolic syndrome and type 2 diabetes have worsened over the past decades. During this time our preventive and therapeutic approach (the 'vaccine'), consisting of a low-fat diet and exercise, has remained fundamentally unchanged. A case is made that these conditions are inter-related and may be caused by a single underlying factor related to the carbohydrate content of diet. The validity of the present approach is challenged when those most knowledgeable in its application succumb to diseases it is meant to prevent. Others argue against the status quo that a low-carbohydrate diet may be more beneficial. A strong belief in the present approach discouraged research into low-carbohydrate diets until recently. Several studies have now demonstrated their benefits and are refuting old claims that they cause harm. Aboriginal people suffer more acutely from the epidemics in question and their dietary history suggests that a sudden increase in carbohydrates is to blame. Recent studies and a case history demonstrate that carbohydrate consumption can drive appetite and over-eating while carbohydrate restriction leads to weight loss and improvement in the markers for metabolic syndrome and type 2 diabetes. The growing evidence in support of low-carbohydrate diets will encounter resistance from economic interests threatened by changes in consumption patterns.


Latino, Asian, and black segregation in U.S. metropolitan areas: are multiethnic metros different?
This study examines 1990 residential segregation levels and 1980-1990 changes in segregation for Latinos, Asians, and blacks in U.S. metropolitan areas. It also evaluates the effect of emerging multiethnic metropolitan area contexts for these segregation patterns. While black segregation levels are still well above those for Latinos and Asians, there is some trend toward convergence over the decade. More than half of the areas increased their Latino segregation levels over the 1980s, and almost three-fourths increased their Asian segregation levels. In contrast, black segregation levels decreased in 88% of metropolitan areas. Multiethnic metropolitan area context is shown to be important for internal segregation dynamics. Black segregation levels are lower, and were more likely to decline in multiethnic metropolitan areas and when other minority groups grew faster than blacks. Latino segregation was also more likely to decline in such areas, and declines in both Latino and Asian segregation were greater when other minority groups were growing. These findings point up the potential for greater mixed-race and mixed-ethnicity coresidence in the neighborhoods of multiethnic metropolitan areas.


Preconception and Diabetes Information (PADI) App for Women with Pregestational Diabetes: a Feasibility and Acceptability Study.
Diabetes mellitus increases the risk of adverse maternal and fetal outcomes. Preconception care is vital to minimise complications; however, preconception care service provision is hindered by inadequate knowledge, resources and care fragmentation. Mobile health technology, particularly smartphone apps, could improve preconception care and pregnancy outcomes for women with diabetes. The aim of this study is to co-create a preconception and diabetes information app with healthcare professionals and women with diabetes and explore the feasibility, acceptability and preliminary effects of the app. A mixed-methods study design employing questionnaires and semi-structured interviews was used to assess preliminary outcome estimates (preconception care knowledge, attitudes and behaviours), and user acceptability. Data analysis included thematic analysis, descriptive statistics and non-parametric tests. Improvements were recorded in knowledge and attitudes to preconception care and patient activation measure following the 3-month app usage. Participants found the app acceptable (satisfaction rating was 72%), useful and informative. The app's usability and usefulness facilitated usage while manual data input and competing priorities were barriers which participants felt could be overcome via personalisation, automation and use of daily reminders. This is the first study to explore the acceptability and feasibility of a preconception and diabetes information app for women with diabetes. Triangulated data suggest that the app has potential to improve preconception care knowledge, attitudes and behaviours. However, in order for women with DM to realise the full potential of the app intervention, particularly improved maternal and fetal outcomes, further development and evaluation is required.


Exhaled volatile organic compounds for phenotyping chronic obstructive pulmonary disease: a cross-sectional study.
Non-invasive phenotyping of chronic respiratory diseases would be highly beneficial in the personalised medicine of the future. Volatile organic compounds can be measured in the exhaled breath and may be produced or altered by disease processes. We investigated whether distinct patterns of these compounds were present in chronic obstructive pulmonary disease (COPD) and clinically relevant disease phenotypes. Breath samples from 39 COPD subjects and 32 healthy controls were collected and analysed using gas chromatography time-of-flight mass spectrometry. Subjects with COPD also underwent sputum induction. Discriminatory compounds were identified by univariate logistic regression followed by multivariate analysis: 1. principal component analysis; 2. multivariate logistic regression; 3. receiver operating characteristic (ROC) analysis. Comparing COPD versus healthy controls, principal component analysis clustered the 20 best-discriminating compounds into four components explaining 71% of the variance. Multivariate logistic regression constructed an optimised model using two components with an accuracy of 69%. The model had 85% sensitivity, 50% specificity and ROC area under the curve of 0.74. Analysis of COPD subgroups showed the method could classify COPD subjects with far greater accuracy. Models were constructed which classified subjects with ≥2% sputum eosinophilia with ROC area under the curve of 0.94 and those having frequent exacerbations 0.95. Potential biomarkers correlated to clinical variables were identified in each subgroup. The exhaled breath volatile organic compound profile discriminated between COPD and healthy controls and identified clinically relevant COPD subgroups. If these findings are validated in prospective cohorts, they may have diagnostic and management value in this disease.


Bounding pixels in computational imaging.
We consider computational imaging problems where we have an insufficient number of measurements to uniquely reconstruct the object, resulting in an ill-posed inverse problem. Rather than deal with this via the usual regularization approach, which presumes additional information which may be incorrect, we seek bounds on the pixel values of the reconstructed image. Formulating the inverse problem as an optimization problem, we find conditions for which a system's measurements can produce a bounded result for both the linear case and the non-negative case (e.g., intensity imaging). We also consider the problem of selecting measurements to yield the most bounded results. Finally we simulate examples of the application of bounded estimation to different two-dimensional multiview systems.


Systematic Review and Meta-analysis of Factors Determining Change to Radical Treatment in Active Surveillance for Localized Prostate Cancer.
Many men with clinically localized prostate cancer are being monitored as part of active surveillance (AS) programs, but little is known about reasons for receiving radical treatment. A systematic review of the evidence about AS was undertaken, with a meta-analysis to identify predictors of radical treatment. A comprehensive search of the Embase, MEDLINE and Web of Knowledge databases to March 2014 was performed. Studies reporting on men with localized prostate cancer followed by AS or monitoring were included. AS was defined where objective eligibility criteria, management strategies, and triggers for clinical review or radical treatment were reported. The 26 AS cohorts included 7627 men, with a median follow-up of 3.5 yr (range of medians 1.5-7.5 yr). The cohorts had a wide range of inclusion criteria, monitoring protocols, and triggers for radical treatment. There were eight prostate cancer deaths and five cases of metastases in 24,981 person-years of follow-up. Each year, 8.8% of men (95% confidence interval 6.7-11.0%) received radical treatment, most commonly because of biopsy findings, prostate-specific antigen triggers, or patient choice driven by anxiety. Studies in which most men changed treatment were those including only low-risk Gleason score 6 disease and scheduled rebiopsies. The wide variety of AS protocols and lack of robust evidence make firm conclusions difficult. Currently, patients and clinicians have to make judgments about the balance of risks and benefits in AS protocols. The publication of robust evidence from randomized trials and longer-term follow-up of cohorts is urgently required. We reviewed 26 studies of men on active surveillance for prostate cancer. There was evidence that studies including men with the lowest risk disease and scheduled rebiopsy had higher rates of radical treatment.


Amplification of the c-myc gene in human medulloblastoma cell lines and xenografts.
Cultured cell lines and xenografts derived from 7 human medulloblastomas were evaluated for amplification of the c-myc, N-myc, epidermal growth factor receptor, and gli genes by Southern blot analysis. Karyotypes of the original biopsies and early passaged cells demonstrated double minute chromosomes in 4 of the 7 cases. All 7 samples (3 cell lines and 4 xenografts) from the 4 tumors with double minute chromosomes contained amplification of the c-myc gene. Cell lines and xenografts derived from the 3 biopsies without double minute chromosomes failed to demonstrate amplification of the 4 genes which were tested, but a rearrangement of the c-myc gene occurred in 1 of the 3 tumors. These observations demonstrate that the c-myc gene is often amplified and/or rearranged in human medulloblastomas and suggest that amplification of this gene provides a growth advantage for medulloblastoma cells in vitro and in vivo.


Monoclonal antibodies to Legionella Mip proteins recognize genus- and species-specific epitopes.
Monoclonal antibodies (MAbs) against the virulence-associated Mip protein of Legionella spp. were raised by immunizing BALB/c mice with (i) Legionella pneumophila, (ii) Legionella micdadei, and (iii) purified recombinant native Mip protein cloned from L. pneumophila Philadelphia 1. Following screening of seeded wells by immunoblot analysis with homologous antigens, eight Mip-specific MAbs were found. These MAbs were chosen to investigate the antigenic diversity of Mip proteins in the genus Legionella. Mip was detected in 82 Legionella strains representing all 34 species tested. One of these MAbs, obtained from immunization with L. micdadei, recognized an epitope common to all Legionella species tested by immunoblot analysis. Another MAb was discovered to be specific for the Mip protein of L. pneumophila. The remaining six MAbs recognized 18 to 79% of Legionella species included in this study. By making use of the MAbs introduced in this study, it could be shown that, based on Mip protein epitope expression, Legionella species can be divided into at least six antigenetically distinct groups. As demonstrated by 43 L. pneumophila strains representing all serogroups, no antigenic diversity of Mip proteins was found for this species. In addition, 18 non-Legionella species, including Chlamydia trachomatis, Neisseria meningitidis, Pseudomonas aeruginosa, and Saccharomyces cerevisiae, all of which are known to carry genes homologous to the Legionella mip genes, were reacted against all eight MAbs. No cross-reactivity was detectable in any of those strains.


Genetic testing including targeted gene panel in a diverse clinical population of children with autism spectrum disorder: Findings and implications.
Genetic testing of children with autism spectrum disorder (ASD) is now standard in the clinical setting, with American College of Medical Genetics and Genomics (ACMGG) guidelines recommending microarray for all children, fragile X testing for boys and additional gene sequencing, including PTEN and MECP2, in appropriate patients. Increasingly, testing utilizing high throughput sequencing, including gene panels and whole exome sequencing, are offered as well. We performed genetic testing including microarray, fragile X testing and targeted gene panel, consistently sequencing 161 genes associated with ASD risk, in a clinical population of 100 well characterized children with ASD. Frequency of rare variants identified in individual genes was compared with that reported in the Exome Aggregation Consortium (ExAC) database. We did not diagnose any conditions with complete penetrance for ASD; however, copy number variants believed to contribute to ASD risk were identified in 12%. Eleven children were found to have likely pathogenic variants on gene panel, yet, after careful analysis, none was considered likely causative of disease. KIRREL3 variants were identified in 6.7% of children compared to 2% in ExAC, suggesting a potential role for KIRREL3 variants in ASD risk. Children with KIRREL3 variants more often had minor facial dysmorphism and intellectual disability. We also observed an increase in rare variants in TSC2. However, analysis of variant data from the Simons Simplex Collection indicated that rare variants in TSC2 occur more commonly in specific racial/ethnic groups, which are more prevalent in our population than in the ExAC database. The yield of genetic testing including microarray, fragile X (boys) and targeted gene panel was 12%. Gene panel did not increase diagnostic yield; however, we found an increase in rare variants in KIRREL3. Our findings reinforce the need for racial/ethnic diversity in large-scale genomic databases used to identify variants that contribute to disease risk.


A genetic defect caused by a triplet repeat expansion in Arabidopsis thaliana.
Variation in the length of simple DNA triplet repeats has been linked to phenotypic variability in microbes and to several human disorders. Population-level forces driving triplet repeat contraction and expansion in multicellular organisms are, however, not well understood. We have identified a triplet repeat-associated genetic defect in an Arabidopsis thaliana variety collected from the wild. The Bur-0 strain carries a dramatically expanded TTC/GAA repeat in the intron of the ISOPROPYL MALATE ISOMERASE LARGE SUB UNIT1 (IIL1; At4g13430) gene. The repeat expansion causes an environment-dependent reduction in IIL1 activity and severely impairs growth of this strain, whereas contraction of the expanded repeat can reverse the detrimental phenotype. The Bur-0 IIL1 defect thus presents a genetically tractable model for triplet repeat expansions and their variability in natural populations.


Ceritinib in ALK-rearranged non-small-cell lung cancer.
Non-small-cell lung cancer (NSCLC) harboring the anaplastic lymphoma kinase gene (ALK) rearrangement is sensitive to the ALK inhibitor crizotinib, but resistance invariably develops. Ceritinib (LDK378) is a new ALK inhibitor that has shown greater antitumor potency than crizotinib in preclinical studies. In this phase 1 study, we administered oral ceritinib in doses of 50 to 750 mg once daily to patients with advanced cancers harboring genetic alterations in ALK. In an expansion phase of the study, patients received the maximum tolerated dose. Patients were assessed to determine the safety, pharmacokinetic properties, and antitumor activity of ceritinib. Tumor biopsies were performed before ceritinib treatment to identify resistance mutations in ALK in a group of patients with NSCLC who had had disease progression during treatment with crizotinib. A total of 59 patients were enrolled in the dose-escalation phase. The maximum tolerated dose of ceritinib was 750 mg once daily; dose-limiting toxic events included diarrhea, vomiting, dehydration, elevated aminotransferase levels, and hypophosphatemia. This phase was followed by an expansion phase, in which an additional 71 patients were treated, for a total of 130 patients overall. Among 114 patients with NSCLC who received at least 400 mg of ceritinib per day, the overall response rate was 58% (95% confidence interval [CI], 48 to 67). Among 80 patients who had received crizotinib previously, the response rate was 56% (95% CI, 45 to 67). Responses were observed in patients with various resistance mutations in ALK and in patients without detectable mutations. Among patients with NSCLC who received at least 400 mg of ceritinib per day, the median progression-free survival was 7.0 months (95% CI, 5.6 to 9.5). Ceritinib was highly active in patients with advanced, ALK-rearranged NSCLC, including those who had had disease progression during crizotinib treatment, regardless of the presence of resistance mutations in ALK. (Funded by Novartis Pharmaceuticals and others; ClinicalTrials.gov number, NCT01283516.).


T-cell receptor gene-modified T cells with shared renal cell carcinoma specificity for adoptive T-cell therapy.
Adoptive therapy with genetically engineered T cells carrying redirected antigen specificity is a new option for the treatment of cancer. This approach is not yet available for metastatic renal cell carcinoma (RCC), due to the scarcity of therapeutically useful reagents. We analyzed tumor-infiltrating lymphocytes (TIL) from RCC to identify T-cell specificities with shared tumor-specific recognition to develop T-cell receptor (TCR)-engineered T lymphocytes for adoptive therapy of RCC. We established a T-cell clone from TIL that recognized a human leukocyte antigen (HLA)-A2-restricted tumor antigen. The TCR alpha- and beta-chain genes were isolated, modified by codon optimization and murinization, and retrovirally transduced into peripheral blood lymphocytes (PBL). A TCR-expressing indicator line (B3Z-TCR53) was established to screen for antigen prevalence in RCC, other malignancies, and normal cell counterparts. TCR53-engineered PBL recapitulated the specificity of the TIL and showed tumor-specific HLA-A2-restricted effector activities (IFN-gamma, tumor necrosis factor-alpha, interleukin-2, macrophage inflammatory protein-1beta, cytotoxicity). PBL-TCR53 of healthy donors and RCC patients exhibited similar transduction efficiency, expansion, and polyfunctional profile. Using B3Z-TCR53 cells, 130 tumor and normal cells were screened and shared TCR53 peptide: MHC expression was found in >60% of RCC and 25% of tumor lines of other histology, whereas normal tissue cells were not recognized. To date, TCR53 is the only TCR with shared HLA-A2-restricted recognition of RCC. It fulfills the criteria for utilization in TCR gene therapy and advances T cell-based immunotherapy to patients with RCC and other malignancies expressing the TCR ligand.


The physiologic responses of Dutch belted rabbits infected with inhalational anthrax.
Bacillus anthracis, the causative agent of anthrax, is a category A priority pathogen that causes extensive damage in humans. For this reason, B. anthracis has been the focus of numerous studies using various animal models. In this study, we explored physiologic parameters in Dutch belted rabbits with inhalation anthrax to characterize the disease progression in this model. To this end, we infected Dutch belted rabbits with 100 LD(50) B. anthracis Ames spores by nasal instillation and continuously recorded various physiologic parameters by using telemetry. In addition, samples were collected at selected times for serum chemistry, hematology, and blood gas analysis. The animals exhibited hemodynamic and respiratory changes that coincided with those reported in human cases of inhalational anthrax infection, including hypotension, altered heart rate, and respiratory distress. Likewise, hematology, serum chemistry, and blood gas analysis revealed trends comparable to human anthrax-related pathophysiology. The Dutch belted rabbit model of inhalational anthrax exhibited most of the physiologic, hematologic, and biochemical sequelae noted in human cases. Therefore, this rabbit model fulfills several of the criteria of a useful animal model for studying disease pathogenesis and evaluating therapeutics during inhalational anthrax.