Comet assay and air-liquid interface exposure system: a new combination to evaluate genotoxic effects of cigarette whole smoke in human lung cell lines.
Over the past three decades, the genotoxic effects of cigarette smoke have generally been evaluated in non-human cell models after exposure to particulate phase, gas phase, or cigarette smoke condensate, rather than the whole smoke aerosol itself. In vitro setups using human cell lines and whole smoke exposure to mimic actual aerosol exposure should more accurately reflect human cigarette smoke exposure. We investigated the VITROCELL® 24 air-liquid interface exposure system in combination with the comet assay to assess DNA damage in two different human lung epithelial cell lines exposed to whole smoke. Results showed a repeatable and reproducible dose-response relationship between DNA damage and increased whole smoke dose in both cell lines. Thus, the combination of the comet assay with the VITROCELL® 24 represents a valuable new in vitro test system to screen and assess DNA damage in human lung cells exposed to whole smoke.


Disparities in combination drug therapy use in older adults with coronary heart disease: a cross-sectional time-series in a nationally representative US sample.
Despite evidence of effective combination drug therapy for secondary prevention of coronary heart disease (CHD), older adults with this condition remain undertreated. To describe time trends (1992-2003) in the adoption of combination cardiac drug therapies (beta-blockers [beta-adrenoceptor antagonists], ACE inhibitors or angiotensin II type 1 receptor antagonists [angiotensin receptor blockers; ARBs], and lipid-lowering agents) among older adults in the US with CHD and to identify factors associated with not using combination therapy. The study took the form of a cross-sectional time-series. The study population consisted of a nationally representative sample of adults aged >or=65 years with CHD (unweighted n = 6331; weighted n = 20.1 million) included in the 1992-2003 Medicare Current Beneficiary Survey. The outcome measure was low-intensity cardiac pharmacotherapy (no drug or single drug therapy with beta-blockers, ACE inhibitors/ARBs or lipid-lowering agents) compared with combination therapy (>or=2 cardiac drugs) for secondary CHD prevention. The use of combination drug therapy in older adults with CHD increased 9-fold during the study period (from 6% in 1992 to 54% in 2003). Adjusted analyses demonstrate that suboptimal drug therapy was independently associated with advanced age (relative risk [RR] 1.18; 95% CI 1.14, 1.23) for patients aged >or=85 years versus patients aged 65-74 years, and with being non-Hispanic Black (RR 1.05; 95% CI 1.01, 1.10) or Hispanic (RR 1.13; 95% CI 1.06, 1.21) versus being non-Hispanic White. Combination drug therapy use for secondary CHD prevention increased in older US adults over the last decade, but improvements were not uniform. The oldest-old, non-Hispanic Blacks and Hispanics experienced slower adoption of optimal medical therapy to improve their long-term prognosis for CHD.


Phorbol ester inhibits bradykinin-stimulated inositol trisphosphate formation and calcium mobilization in neuroblastoma x glioma hybrid NG108-15 cells.
In neuroblastoma x glioma hybrid NG108-15 cells, bradykinin (BK) receptor stimulation leads to phosphoinositide hydrolysis, formation of inositol phosphates and mobilization of intracellular calcium. Treatment of the cells with 12-O-tetradecanoyl phorbol 13-acetate (TPA) suppressed the spike phase of increases in intracellular calcium concentration. In radioligand binding studies, TPA treatment did not interfere with [3H]BK specific binding to intact cells or to cell membranes. The ability of guanyl-5'-yl-imidodiphosphate to promote the conversion of the high affinity sites of the BK receptors into a low affinity sites was unaffected by TPA. TPA treatment showed the dose-dependent, noncompetitive inhibition of BK-stimulated formation of inositol trisphosphate. In the membrane preparations from TPA-treated cells, guanosine 5'-(3-O-thio)triphosphate-stimulated inositol trisphosphate formation was inhibited by 50%. These data indicate that TPA exerts its inhibitory action on BK responses at the sites of guanine nucleotide-binding protein or phospholipase C or both.


The T8590C polymorphism of CYP4A11 and 20-hydroxyeicosatetraenoic acid in essential hypertension.
A role for a deficit in transport actions of 20-hydroxyeicosatetraenoic acid (20-HETE) in hypertension is supported by the following: (1) diminished renal 20-HETE in Dahl-S rats; (2) altered salt- and furosemide-induced 20-HETE responses in salt-sensitive hypertensive subjects; and (3) increased population risk for hypertension in C allele carriers of the T8590C polymorphism of CYP4A11, which encodes an enzyme with reduced catalytic activity. We determined T8590C genotypes in 32 hypertensive subjects, 25 of whom were phenotyped for salt sensitivity of blood pressure and insulin sensitivity. Urine 20-HETE was lowest in insulin-resistant, salt-sensitive subjects (F=5.56; P<0.02). Genotypes were 13 TT, 2 CC, and 17 CT. C allele frequency was 32.8% (blacks: 38.9%; whites: 25.0%). C carriers (CC+CT) and TT subjects were similarly distributed among salt- and insulin-sensitivity phenotypes. C carriers had higher diastolic blood pressures and aldosterone:renin and waist:hip ratios but lower furosemide-induced fractional excretions of Na and K than TT. The T8590C genotype did not relate to sodium balance or pressure natriuresis. However, C carriers, compared with TT, had diminished 20-HETE responses to salt loading after adjustment for serum insulin concentration and resetting of the negative relationship between serum insulin and urine 20-HETE to a 1-microg/h lower level of 20-HETE. The effect of C was insulin independent and equipotent to 18 microU/mL of insulin (Delta20-HETE= 2.84-0.054xinsulin-0.98xC; r(2)=0.53; F=11.1; P<0.001). Hence, genetic (T8590C) and environmental (insulin) factors impair 20-HETE responses to salt in human hypertension. We propose that genotype analyses with sufficient homozygous CC will establish definitive relationships among 20-HETE, salt sensitivity of blood pressure, and insulin resistance.


Prognostic implications of angiographically normal and insignificantly narrowed coronary arteries.
The clinical presentation and prognosis of 1,977 consecutive patients with normal coronary arteries or "insignificant" coronary artery disease (CAD) (no major epicardial artery with 75% or more luminal diameter narrowing) were examined. Compared with patients with significant CAD, these patients had a lower frequency of traditional cardiac risk factors and abnormalities on the rest and exercise electrocardiogram. Cardiac survival was 99% at 5 years of follow-up and 98% at 10 years for patients with normal or insignificantly narrowed coronary arteries. Patients with normal coronary arteries differed from those with insignificant CAD in their myocardial infarction free survival rate: 99% at 5 years and 98% at 10 years for patients with normal coronary arteries, compared with 97% at 5 years and 90% at 10 years for patients with insignificant CAD. A strong relation occurred between the amount of insignificant CAD and follow-up cardiac events (chi 2 = 21.5, p less than 0.0001). Cardiac risk factors were statistically related to the risk of follow-up cardiovascular events when considered alone (chi 2 = 4.93, p = 0.026), but this relation lost significance after adjusting for the effect of coronary anatomy. Patients in both groups continued to have cardiac symptoms that resulted in frequent hospitalizations, medication use and job disability. Almost 50% in any given year of follow-up could not perform activities of high metabolic equivalent requirement and 70% had continuing symptoms of chest discomfort. Although these patients are at low risk of death, many remain functionally impaired for years.


Deformation measurements by digital image correlation with automatic merging of data distributed in time.
This paper presents a method to analyze 3D displacement data captured by digital image correlation (DIC) method over a long period of time. It allows monitoring of an object when a 3D DIC setup is not fixed in the same position between a consecutive series of measurements. An implementation of the data merging procedure is described and a proof of concept is provided using example measurements for both a numerical model and a real experiment in laboratory conditions. We evaluated the accuracy and discuss the main sources of errors. The obtained results prove the method is feasible for in situ long-term measurements and monitoring in industry and civil engineering.


A CCD based focal spot camera.
Rapid, quantative assessment of x-radiation source intensity distributions would be useful in several areas of radiation physics. A simple x-ray sensitive CCD based camera for the measurement of focal spot and radiation beam intensity profiles has been constructed. Focal spot images recorded with the CCD camera and radiographic emulsion are compared and the advantage of the CCD camera in rapidly recording true radiation intensity distributions is demonstrated.


Functional Conservation in the SIAMESE-RELATED Family of Cyclin-Dependent Kinase Inhibitors in Land Plants.
The best-characterized members of the plant-specific SIAMESE-RELATED (SMR) family of cyclin-dependent kinase inhibitors regulate the transition from the mitotic cell cycle to endoreplication, also known as endoreduplication, an altered version of the cell cycle in which DNA is replicated without cell division. Some other family members are implicated in cell cycle responses to biotic and abiotic stresses. However, the functions of most SMRs remain unknown, and the specific cyclin-dependent kinase complexes inhibited by SMRs are unclear. Here, we demonstrate that a diverse group of SMRs, including an SMR from the bryophyte Physcomitrella patens, can complement an Arabidopsis thaliana siamese (sim) mutant and that both Arabidopsis SIM and P. patens SMR can inhibit CDK activity in vitro. Furthermore, we show that Arabidopsis SIM can bind to and inhibit both CDKA;1 and CDKB1;1. Finally, we show that SMR2 acts to restrict cell proliferation during leaf growth in Arabidopsis and that SIM, SMR1/LGO, and SMR2 play overlapping roles in controlling the transition from cell division to endoreplication during leaf development. These results indicate that differences in SMR function in plant growth and development are primarily due to differences in transcriptional and posttranscriptional regulation, rather than to differences in fundamental biochemical function.


Genetic basis of the formation and identity of type I and type II neurons in Drosophila embryos.
The embryonic peripheral nervous system of Drosophila contains two main types of sensory neurons: type I neurons, which innervate external sense organs and chordotonal organs, and type II multidendritic neurons. Here, we analyse the origin of the difference between type I and type II in the case of the neurons that depend on the proneural genes of the achaete-scute complex (ASC). We show that, in Notch- embryos, the type I neurons are missing while type II neurons are produced in excess, indicating that the type I/type II choice relies on Notch-mediated cell communication. In contrast, both type I and type II neurons are absent in numb- embryos and after ubiquitous expression of tramtrack, indicating that the activity of numb and the absence of tramtrack are required to produce both external sense organ and multidendritic neural fates. The analysis of string- embryos reveals that when the precursors are unable to divide they differentiate mostly into type II neurons, indicating that the type II is the default neuronal fate. We also report a new mutant phenotype where the ASC-dependent neurons are converted into type II neurons, providing evidence for the existence of one or more genes required for maintaining the alternative (type I) fate. Our results suggest that the same mechanism of type I/type II specification may operate at a late step of the ASC-dependent lineages, when multidendritic neurons arise as siblings of the external sense organ neurons and, at an early step, when other multidendritic neurons precursors arise as siblings of external sense organ precursors.


Calcium-permeable AMPA receptors are expressed in a rodent model of status epilepticus.
A study was undertaken to characterize the plasticity of AMPA receptor (AMPAR)-mediated neurotransmission in the hippocampus during status epilepticus (SE). SE was induced by pilocarpine, and animals were studied 10 minutes (refractory SE) or 60 minutes (late SE) after the onset of the first grade 5 seizures. AMPAR-mediated currents were recorded from CA1 pyramidal neurons and dentate granule cells (DGCs) by voltage clamp technique. The surface expression of GluA2 subunit on hippocampal membranes was determined using a biotinylation assay. GluA2 internalization and changes in intracellular calcium ([Ca](i)) levels were studied in hippocampal cultures using immunocytochemical and live-imaging techniques. AMPAR antagonist treatment of SE was evaluated by video and electroencephalography. AMPAR-mediated currents recorded from CA1 neurons from refractory and late SE animals were inwardly rectifying, and philanthotoxin-sensitive; similar changes were observed in recordings obtained from DGCs from refractory SE animals. GluA2 subunit surface expression was reduced in the hippocampus during refractory and late SE. In cultured hippocampal pyramidal neurons, recurrent bursting diminished surface expression of the GluA2 subunit and enhanced its internalization rate. Recurrent bursting-induced increase in [Ca](i) levels was reduced by selective inhibition of GluA2-lacking AMPARs. GYKI-52466 terminated diazepam-refractory SE. During SE, there is rapid, ongoing plasticity of AMPARs with the expression of GluA2-lacking AMPARs. These receptors provide another source of Ca(2+) entry into the principal neurons. Benzodiazepam-refractory SE can be terminated by AMPAR antagonism. The data identify AMPARs as a potential therapeutic target for the treatment of SE.


Facilitated communication and authorship: a systematic review.
Facilitated Communication (FC) is a technique whereby individuals with disabilities and communication impairments allegedly select letters by typing on a keyboard while receiving physical support, emotional encouragement, and other communication supports from facilitators. The validity of FC stands or falls on the question of who is authoring the typed messages--the individual with a disability or the facilitator. The International Society for Augmentative and Alternative Communication (ISAAC) formed an Ad Hoc Committee on FC and charged this committee to synthesize the evidence base related to this question in order to develop a position statement. The purpose of this paper is to report this synthesis of the extant peer-reviewed literature on the question of authorship in FC. A multi-faceted search was conducted including electronic database searches, ancestry searches, and contacting selected authors. The authors considered synopses of systematic reviews, and systematic reviews, which were supplemented with individual studies not included in any prior reviews. Additionally, documents submitted by the membership were screened for inclusion. The evidence was classified into articles that provided (a) quantitative experimental data related to the authorship of messages, (b) quantitative descriptive data on the output generated through FC without testing of authorship, (c) qualitative descriptive data on the output generated via FC without testing of authorship, and (d) anecdotal reports in which writers shared their perspectives on FC. Only documents with quantitative experimental data were analyzed for authorship. Results indicated unequivocal evidence for facilitator control: messages generated through FC are authored by the facilitators rather than the individuals with disabilities. Hence, FC is a technique that has no validity.


Quantitative analysis of the debonding structure of soft adhesives.
We experimentally investigate the growth dynamics of cavities nucleating during the first stages of debonding of three different model adhesives. The material properties of these adhesives range from a more liquid-like material to a soft viscoelastic solid and are carefully characterized by small strain oscillatory shear rheology as well as large strain uniaxial extension. The debonding experiments are performed on a probe tack set-up. Using high contrast images of the debonding process and precise image analysis tools, we quantify the total projected area of the cavities, the average cavity shape and growth rate and link these observations to the material properties. These measurements are then used to access corrected effective stress and strain curves that can be directly compared to the results from the uniaxial extension.


Two-dimensional depth-resolved Mueller matrix characterization of biological tissue by optical coherence tomography.
We built a polarization-sensitive optical coherence tomographic system and measured the two-dimensional depth-resolved full 4 x 4 Mueller matrix of biological tissue for what is believed to be the first time. The Mueller matrix measurements, which we made by varying the polarization states of the light source and the detector, yielded a complete characterization of the polarization property of the tissue sample. The initial experimental results indicated that this new approach reveals some tissue structures that are not perceptible in standard optical coherence tomography.


Minimum aerosol layer detection sensitivities and their subsequent impacts on aerosol optical thickness retrievals in CALIPSO level 2 data products.
Due to instrument sensitivities and algorithm detection limits, level 2 (L2) Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) 532nm aerosol extinction profile retrievals are often populated with retrieval fill values (RFVs), which indicate the absence of detectable levels of aerosol within the profile. In this study, using 4 years (2007-2008 and 2010-2011) of CALIOP version 3 L2 aerosol data, the occurrence frequency of daytime CALIOP profiles containing all RFVs (all-RFV profiles) is studied. In the CALIOP data products, the aerosol optical thickness (AOT) of any all-RFV profile is reported as being zero, which may introduce a bias in CALIOP-based AOT climatologies. For this study, we derive revised estimates of AOT for all-RFV profiles using collocated Moderate Resolution Imaging Spectroradiometer (MODIS) Dark Target (DT) and, where available, AErosol RObotic NEtwork (AERONET) data. Globally, all-RFV profiles comprise roughly 71% of all daytime CALIOP L2 aerosol profiles (i.e., including completely attenuated profiles), accounting for nearly half (45 %) of all daytime cloud-free L2 aerosol profiles. The mean collocated MODIS DT (AERONET) 550 nm AOT is found to be near 0.06 (0.08) for CALIOP all-RFV profiles. We further estimate a global mean aerosol extinction profile, a so-called "noise floor", for CALIOP all-RFV profiles. The global mean CALIOP AOT is then recomputed by replacing RFV values with the derived noise-floor values for both all-RFV and non-all-RFV profiles. This process yields an improvement in the agreement of CALIOP and MODIS over-ocean AOT.


Cell signaling of angiotensin II on vascular tone: novel mechanisms.
Angiotensin II (Ang II) is a pleiotropic hormone that influences the function of many cell types and regulates many organ systems. In the cardiovascular system, it is a potent vasoconstrictor that increases peripheral vascular resistance and elevates arterial pressure. It also promotes inflammation, hypertrophy, and fibrosis, which are important in vascular remodeling in cardiovascular diseases. The diverse actions of Ang II are mediated via AT(1) and AT(2) receptors, which couple to many signaling molecules, including small G proteins, phospholipases, mitogen-activated protein (MAP) kinases, phosphatases, tyrosine kinases, NADPH oxidase, and transcription factors. In general, acute Ang II stimulation induces vasoconstriction through changes in the intracellular free calcium concentration [Ca(2+)](i), whereas long-term stimulation leads to cell proliferation and proinflammatory responses. This review focuses on signaling processes of vasoconstriction and highlights some new mechanisms regulating the contractile machinery in controlling vasomotor tone by Ang II, including RhoA/Rho kinase, transient receptor potential (TRP) channels, reactive oxygen species, and arachidonic acid metabolites.


Modern treatment of gastric gastrointestinal stromal tumors.
Gastrointestinal stromal tumors (GIST) are rare mesenchymal smooth muscle sarcomas that can arise anywhere within the gastrointestinal tract. Sporadic mutations within the tyrosine kinase receptors of the interstitial cells of Cajal have been identified as the key molecular step in GIST carcinogenesis. Although many patients are asymptomatic, the most common associated symptoms include: abdominal pain, dyspepsia, gastric outlet obstruction, and anorexia. Rarely, GIST can perforate causing life-threatening hemoperitoneum. Most are ultimately diagnosed on cross-sectional imaging studies (i.e., computed tomography and/or magnetic resonance imaging in combination with upper endoscopy. Endoscopic ultrasonographic localization of these tumors within the smooth muscle layer and acquisition of neoplastic spindle cells harboring mutations in the c-KIT gene is pathognomonic. Curative treatment requires a complete gross resection of the tumor. Both open and minimally invasive operations have been shown to reduce recurrence rates and improve long-term survival. While there is considerable debate over whether GIST can be benign neoplasms, we believe that all GIST have malignant potential, but vary in their propensity to recur after resection and metastasize to distant organ sites. Prognostic factors include location, size (i.e., > 5 cm), grade (> 5-10 mitoses per 50 high power fields and specific mutational events that are still being defined. Adjuvant therapy with tyrosine kinase inhibitors, such as imatinib mesylate, has been shown to reduce the risk of recurrence after one year of therapy. Treatment of locally-advanced or borderline resectable gastric GIST with neoadjuvant imatinib has been shown to induce regression in a minority of patients and stabilization in the majority of cases. This treatment strategy potentially reduces the need for more extensive surgical resections and increases the number of patients eligible for curative therapy. The modern surgical treatment of gastric GIST combines the novel use of targeted therapy and aggressive minimally invasive surgical procedures to provide effective treatment for this lethal, but rare gastrointestinal malignancy.


In vitro apoptotic response of freshly isolated chronic myeloid leukemia cells to all-trans retinoic acid and cytosine arabinoside.
Chronic myeloid leukemia (CML) is a hematological malignancy resulting from clonal expansion and massive accumulation of leukemic myeloid cells that retain differentiation and maturation capacity. Since CML cell accumulation has been related to apoptosis inhibition by the product of the BCR-ABL gene, attempts to eradicate leukemic cells would require therapeutic drugs able to overcome this inherent resistance. Here, we investigated in vitro the apoptotic effect of all-trans retinoic acid (ATRA) and cytosine arabinoside (ARA-C), employed alone, in combination or in sequence, on freshly isolated cells from 10 patients with chronic-phase CML. Our cell cultures showed that both ATRA and ARA-C were able to induce apoptosis in CML cells, even if ARA-C resulted more effective than ATRA. The combined use of ATRA and ARA-C seemed to have only an additive effect while the sequential use did not show any advantage. These in vitro observations indicate that ATRA and ARA-C may be effective in reducing CML cells through apoptosis induction, suggesting that it could be worthwhile to examine ATRA and ARA-C combinations in the therapy of CML.


Lactobacillus reuteri in management of Helicobacter pylori infection in dyspeptic patients: a double-blind placebo-controlled randomized clinical trial.
The eradication rate of Helicobacter pylori following the standard triple therapy is declining. This study was conducted to test whether the addition of Lactobacillus reuteri to the standard triple therapy improves the eradication rates as well as the clinical and pathological aspects in H. pylori infection. A total of 70 treatment-naïve patients were randomly assigned into group A (the L. reuteri treated group) and group B (the placebo control group). Patients were treated by the standard triple therapy for 2 weeks and either L. reuteri or placebo for 4 weeks. They were examined by symptom questionnaire, H. pylori antigen in stool, upper endoscopy with biopsies for rapid urease test and histopathological examination before treatment and 4 weeks after treatment. The eradication rate of H. pylori infection was 74.3% and 65.7% for both L. reuteri and placebo treated groups, respectively. There was a significant difference regarding the reported side effects, where patients treated with L. reuteri reported less diarrhea and taste disorders than placebo group. A significant difference within each group was observed after treatment regarding Gastrointestinal Symptom Rating Scale (GSRS) scores; patients treated with L. reuteri showed more improvement of gastrointestinal symptoms than the placebo treated group. The severity and activity of H. pylori associated gastritis were reduced after 4 weeks of therapy in both groups. The L. reuteri treated group showed significant improvement compared with the placebo treated group. Triple therapy of H. pylori supplemented with L. reuteri increased eradication rate by 8.6%, improved the GSRS score, reduced the reported side effects and improved the histological features of H. pylori infection when compared with placebo-supplemented triple therapy.


Heterogeneity among 16S-23S rRNA intergenic spacers of species within the 'Streptococcus milleri group'.
The 16S-23S rRNA intergenic spacer has been suggested as a suitable region of the bacterial genome from which to derive useful taxonomic information, particularly with regard to identification at the species level. To investigate this approach as an aid to the identification of the three species comprising the 'Streptococcus milleri group' (SMG), the spacers of isolates of Streptococcus intermedius, Streptococcus anginosus and Streptococcus constellatus were amplified by PCR and length polymorphisms determined by agarose gel electrophoresis. Phenotypically atypical isolates which had been identified presumptively as belonging to these three species were also included. Spacers from two representatives of each spacer length found within the three SMG species were sequenced. 16S-23S rRNA intergenic spacer length polymorphisms allowed discrimination between S. anginosus (350 bp or 450 bp amplification product) and S. constellatus (380 bp amplification product), species that are difficult to differentiate phenotypically. S. intermedius (330 bp or 450 bp amplification product) and S. anginosus (350 bp or 450 bp amplification product) were not reliably distinguished by this method but are phenotypically distinct. Sequencing data demonstrated that the spacers had a central region of highly variable length flanked by conserved regions which included a single tRNA(Ala) gene. Polymorphism in the length of the 16S-23S spacer determined by PCR provides a rapid and useful adjunct to strain identification for S. anginosus and S. constellatus, which are not readily differentiated phenotypically.


Integrating survey and molecular approaches to better understand wildlife disease ecology.
Infectious wildlife diseases have enormous global impacts, leading to human pandemics, global biodiversity declines and socio-economic hardship. Understanding how infection persists and is transmitted in wildlife is critical for managing diseases, but our understanding is limited. Our study aim was to better understand how infectious disease persists in wildlife populations by integrating genetics, ecology and epidemiology approaches. Specifically, we aimed to determine whether environmental or host factors were stronger drivers of Salmonella persistence or transmission within a remote and isolated wild pig (Sus scrofa) population. We determined the Salmonella infection status of wild pigs. Salmonella isolates were genotyped and a range of data was collected on putative risk factors for Salmonella transmission. We a priori identified several plausible biological hypotheses for Salmonella prevalence (cross sectional study design) versus transmission (molecular case series study design) and fit the data to these models. There were 543 wild pig Salmonella observations, sampled at 93 unique locations. Salmonella prevalence was 41% (95% confidence interval [CI]: 37-45%). The median Salmonella DICE coefficient (or Salmonella genetic similarity) was 52% (interquartile range [IQR]: 42-62%). Using the traditional cross sectional prevalence study design, the only supported model was based on the hypothesis that abundance of available ecological resources determines Salmonella prevalence in wild pigs. In the molecular study design, spatial proximity and herd membership as well as some individual risk factors (sex, condition score and relative density) determined transmission between pigs. Traditional cross sectional surveys and molecular epidemiological approaches are complementary and together can enhance understanding of disease ecology: abundance of ecological resources critical for wildlife influences Salmonella prevalence, whereas Salmonella transmission is driven by local spatial, social, density and individual factors, rather than resources. This enhanced understanding has implications for the control of diseases in wildlife populations. Attempts to manage wildlife disease using simplistic density approaches do not acknowledge the complexity of disease ecology.


A generic framework to simulate realistic lung, liver and renal pathologies in CT imaging.
Realistic three-dimensional (3D) mathematical models of subtle lesions are essential for many computed tomography (CT) studies focused on performance evaluation and optimization. In this paper, we develop a generic mathematical framework that describes the 3D size, shape, contrast, and contrast-profile characteristics of a lesion, as well as a method to create lesion models based on CT data of real lesions. Further, we implemented a technique to insert the lesion models into CT images in order to create hybrid CT datasets. This framework was used to create a library of realistic lesion models and corresponding hybrid CT images. The goodness of fit of the models was assessed using the coefficient of determination (R(2)) and the visual appearance of the hybrid images was assessed with an observer study using images of both real and simulated lesions and receiver operator characteristic (ROC) analysis. The average R(2) of the lesion models was 0.80, implying that the models provide a good fit to real lesion data. The area under the ROC curve was 0.55, implying that the observers could not readily distinguish between real and simulated lesions. Therefore, we conclude that the lesion-modeling framework presented in this paper can be used to create realistic lesion models and hybrid CT images. These models could be instrumental in performance evaluation and optimization of novel CT systems.


SU-E-T-515: Comparison of Two Dose-Specification Methods for the Treatment of Uterine Cervix Cancer with HDR Brachytherapy Using a Tandem and Ring Applicator.
To compare the dose to the high-risk clinical target volume (HRCTV)-D90 and dose-limiting structures with the dose prescription to point A (Manchester system) using conventional methods to image-guided treatment planning with dose optimization and a dose prescription to D90 of the HRCTV. CT-MR-fused image sets for 16 patients (stage IA2-IIIB) with uterine cervix cancer were obtained for each of 5 fractions of HDR brachytherapy using tandem and ring applicators to generate 2 sets of plans. In the first set, the dose was prescribed to point A with a standard dwell weight pattern reflecting the Manchester system loading pattern. In the second set, the dose was prescribed to D90 of the HRCTV with graphical plan optimization. Parameters of clinical significance (HRCTV-D90 and dose received by 1.0, 2.0, and 5.0cc of the bladder, rectum, and sigmoid) were compared, as recommended by GEC-ESTRO. HRCTV-V for 6 patients was =30cc (largest radius, <2.0cm) and >30cc (largest radius, =2cm) for the others. For the first set of plans, when HRCTV-V was <30cc, the ratio of doses HRCTV-D90/dose to point A varied from 1.10 to 1.40. When HRCTV-V was >30cc, it varied from 0.75 to 0.95. In the second set of plans, HRCTV-D90 always received the prescription dose. Doses to 2cc of one or more organs at risk exceeded >10% of the limiting doses for 9 patients in the first set of plans but remained <10% in all patients in the second set of plans. A 10-25% dose reduction to 2cc of the bladder and 15-35% to 2cc of the rectum and sigmoid occurred in the second set of plans. Image-guided brachytherapy with dose optimization yields a prescribed dose to the HRCTV-D90 while maintaining doses to organs at risk close to their limiting value, offering a dosimetric benefit over the traditional method.


Dual modes of natural selection on upstream open reading frames.
Upstream open reading frames (uORFs) are common features of eukaryotic genes, occurring in 10%-25% of 5' leader sequences. Upstream ORFs that have been subjected to experimental analysis have been generally found to decrease translational efficiency of the downstream coding sequence. Previous investigations of uORFs in mammals and yeast have detected uORFs conserved over long evolutionary distances, prompting speculation about the nature and cause of the natural selection underlying such conservation. We have analyzed uORFs in the basidiomycetous fungal pathogen Cryptococcus neoformans to discern the properties of this purifying selection. We find that uORFs in the Cryptococcus species complex are conserved at twice the expected rate, and we report 122 uORFs that are conserved among all four sequenced Cryptococcus strains. A significantly greater proportion of uORF losses occur via direct mutation to the uORF start codon than expected. This observation suggests that mutational disruption of a uORF that leaves the start codon intact may be selectively disadvantageous, perhaps because of the risk of premature translation initiation. Accounting for this constrained mode of loss and comparing the relative conservation of uORFs between the 5' leader and control sequences enables us to calculate that at least a third of uORFs may be conserved for their effects on translational efficiency. The remaining fraction may be conserved either by chance or as a result of selective pressure to prevent premature translation initiation from the uORF start codon. We find that the majority of conserved uORFs do not exhibit codon usage bias or conservation at the amino acid level, and therefore they do not likely encode bioactive peptides. Our analysis suggests that uORFs are an important and underappreciated mechanism of post-transcriptional gene regulation in eukaryotes.


Machining learning predicts the need for escalated care and mortality in COVID-19 patients from clinical variables.
Objective: This study aimed to develop a machine learning algorithm to identify key clinical measures to triage patients more effectively to general admission versus intensive care unit (ICU) admission and to predict mortality in COVID-19 pandemic. Materials and methods: This retrospective study consisted of 1874 persons-under-investigation for COVID-19 between February 7, 2020, and May 27, 2020 at Stony Brook University Hospital, New York. Two primary outcomes were ICU admission and mortality compared to COVID-19 positive patients in general hospital admission. Demographic, vitals, symptoms, imaging findings, comorbidities, and laboratory tests at presentation were collected. Predictions of mortality and ICU admission were made using machine learning with 80% training and 20% testing. Performance was evaluated using receiver operating characteristic (ROC) area under the curve (AUC). Results: A total of 635 patients were included in the analysis (age 60±11, 40.2% female). The top 6 mortality predictors were age, procalcitonin, C-creative protein, lactate dehydrogenase, D-dimer and lymphocytes. The top 6 ICU admission predictors are procalcitonin, lactate dehydrogenase, C-creative protein, pulse oxygen saturation, temperature and ferritin. The best machine learning algorithms predicted mortality with 89% AUC and ICU admission with 79% AUC. Conclusion: This study identifies key independent clinical parameters that predict ICU admission and mortality associated with COVID-19 infection. The predictive model is practical, readily enhanced and retrained using additional data. This approach has immediate translation and may prove useful for frontline physicians in clinical decision making under time-sensitive and resource-constrained environment.


Structured nucleosome fingerprints enable high-resolution mapping of chromatin architecture within regulatory regions.
Transcription factors canonically bind nucleosome-free DNA, making the positioning of nucleosomes within regulatory regions crucial to the regulation of gene expression. Using the assay of transposase accessible chromatin (ATAC-seq), we observe a highly structured pattern of DNA fragment lengths and positions around nucleosomes in Saccharomyces cerevisiae, and use this distinctive two-dimensional nucleosomal "fingerprint" as the basis for a new nucleosome-positioning algorithm called NucleoATAC. We show that NucleoATAC can identify the rotational and translational positions of nucleosomes with up to base-pair resolution and provide quantitative measures of nucleosome occupancy in S. cerevisiae, Schizosaccharomyces pombe, and human cells. We demonstrate the application of NucleoATAC to a number of outstanding problems in chromatin biology, including analysis of sequence features underlying nucleosome positioning, promoter chromatin architecture across species, identification of transient changes in nucleosome occupancy and positioning during a dynamic cellular response, and integrated analysis of nucleosome occupancy and transcription factor binding.


Population diversity in Pacific herring of the Puget Sound, USA.
Demographic, functional, or habitat diversity can confer stability on populations via portfolio effects (PEs) that integrate across multiple ecological responses and buffer against environmental impacts. The prevalence of these PEs in aquatic organisms is as yet unknown, and can be difficult to quantify; however, understanding mechanisms that stabilize populations in the face of environmental change is a key concern in ecology. Here, we examine PEs in Pacific herring (Clupea pallasii) in Puget Sound (USA) using a 40-year time series of biomass data for 19 distinct spawning population units collected using two survey types. Multivariate auto-regressive state-space models show independent dynamics among spawning subpopulations, suggesting that variation in herring production is partially driven by local effects at spawning grounds or during the earliest life history stages. This independence at the subpopulation level confers a stabilizing effect on the overall Puget Sound spawning stock, with herring being as much as three times more stable in the face of environmental perturbation than a single population unit of the same size. Herring populations within Puget Sound are highly asynchronous but share a common negative growth rate and may be influenced by the Pacific Decadal Oscillation. The biocomplexity in the herring stock shown here demonstrates that preserving spatial and demographic diversity can increase the stability of this herring population and its availability as a resource for consumers.


Improving inferences about private land conservation by accounting for incomplete reporting.
Private lands provide key habitat for imperiled species and are core components of function protectected area networks; yet, their incorporation into national and regional conservation planning has been challenging. Identifying locations where private landowners are likely to participate in conservation initiatives can help avoid conflict and clarify trade-offs between ecological benefits and sociopolitical costs. Empirical, spatially explicit assessment of the factors associated with conservation on private land is an emerging tool for identifying future conservation opportunities. However, most data on private land conservation are voluntarily reported and incomplete, which complicates these assessments. We used a novel application of occupancy models to analyze the occurrence of conservation easements on private land. We compared multiple formulations of occupancy models with a logistic regression model to predict the locations of conservation easements based on a spatially explicit social-ecological systems framework. We combined a simulation experiment with a case study of easement data in Idaho and Montana (United States) to illustrate the utility of the occupancy framework for modeling conservation on private land. Occupancy models that explicitly accounted for variation in reporting produced estimates of predictors that were substantially less biased than estimates produced by logistic regression under all simulated conditions. Occupancy models produced estimates for the 6 predictors we evaluated in our case study that were larger in magnitude, but less certain than those produced by logistic regression. These results suggest that occupancy models result in qualitatively different inferences regarding the effects of predictors on conservation easement occurrence than logistic regression and highlight the importance of integrating variable and incomplete reporting of participation in empirical analysis of conservation initiatives. Failure to do so can lead to emphasizing the wrong social, institutional, and environmental factors that enable conservation and underestimating conservation opportunities in landscapes where social norms or institutional constraints inhibit reporting.


Disaster preparedness for vulnerable persons receiving in-home, long-term care in South Carolina.
The purpose of this study was to examine how agencies in South Carolina that provide in-home health care and personal care services help older and/or disabled clients to prepare for disasters. The study also examines how agencies safeguard clients' records, train staff, and how they could improve their preparedness. The relevant research and practice literature was reviewed. Nine public officials responsible for preparedness for in-home health care and personal care services in South Carolina were interviewed. A telephone survey instrument was developed that was based on these interviews and the literature review. Administrators from 16 agencies that provide in-home personal care to 2,147 clients, and five agencies that provide in-home health care to 2,180 clients, were interviewed. Grounded theory analysis identified major themes in the resulting qualitative data; thematic analysis organized the content. Federal regulations require preparedness for agencies providing in-home health care ("home health"). No analogous regulations were found for in-home personal care. The degree of preparedness varied substantially among personal care agencies. Most personal care agencies were categorized as "less" prepared or "moderately" prepared. The findings for agencies in both categories generally suggest lack of preparedness in: (1) identifying clients at high risk and assisting them in planning; (2) providing written materials and/or recommendations; (3) protecting records; (4) educating staff and clients; and (5) coordinating disaster planning and response across agencies. Home health agencies were better prepared than were personal care agencies. However, some home health administrators commented that they were unsure how well their plans would work during a disaster, given a lack of training. The majority of home health agency administrators spoke of a need for better coordination and/or more preparedness training. Agencies providing personal care and home health services would benefit from developing stronger linkages with their local preparedness systems. The findings support incorporating disaster planning in the certification requirements for home health agencies, and developing additional educational resources for administrators and staff of personal care agencies and their clients.


Development of a Chromatic Pupillography Protocol for the First Gene Therapy Trial in Patients With CNGA3-Linked Achromatopsia.
To establish a feasible and sensitive pupillographic protocol to assess outer and inner retinal function for the first gene therapy trial in achromatopsia patients (ACHM) with mutations in CNGA3. Twenty-seven CNGA3-ACHM patients and 22 age-matched control subjects were tested using chromatic pupillography. Three different protocols were established to assess the pupillary light reflex parameters and to create the final protocol. In the individual protocols, various stimulus parameters (i.e., intensity, duration, wavelength, adaptation states) were applied to evaluate the impact of these stimuli on the pupillary response in untreated ACHM patients. In the light-adapted conditions, CNGA3-ACHM patients showed significantly reduced maximal amplitudes compared with the control group when using a 1-second high intensity (28-lux corneal illumination) blue or red stimulus (P < 0.005). In the dark-adapted conditions, CNGA3-ACHM patients unexpectedly revealed significantly increased maximal amplitudes when stimulating with red (1 second) or blue (4 ms and 1 second) stimuli of low intensity (0.01-lux corneal illumination; P < 0.05). Pupil responses of CNGA3-ACHM patients after high intensity (28 lux) red and blue 1-second stimuli were within the normal range. Chromatic pupillography demonstrated significant reduced pupil responses to stimuli addressing primarily cone function, an increased sensitivity to rod-favoring stimuli and evidence for disinhibition of intrinsically photosensitive retinal ganglion cells in CNGA3-ACHM patients. A final protocol was established based on these findings. These conclusions may be useful for the objective assessment of efficacy gained by gene therapy or other innovative interventions in this hereditary retinal disorder.


A genome-wide association study identifies pancreatic cancer susceptibility loci on chromosomes 13q22.1, 1q32.1 and 5p15.33.
We conducted a genome-wide association study of pancreatic cancer in 3,851 affected individuals (cases) and 3,934 unaffected controls drawn from 12 prospective cohort studies and 8 case-control studies. Based on a logistic regression model for genotype trend effect that was adjusted for study, age, sex, self-described ancestry and five principal components, we identified eight SNPs that map to three loci on chromosomes 13q22.1, 1q32.1 and 5p15.33. Two correlated SNPs, rs9543325 (P = 3.27 x 10(-11), per-allele odds ratio (OR) 1.26, 95% CI 1.18-1.35) and rs9564966 (P = 5.86 x 10(-8), per-allele OR 1.21, 95% CI 1.13-1.30), map to a nongenic region on chromosome 13q22.1. Five SNPs on 1q32.1 map to NR5A2, and the strongest signal was at rs3790844 (P = 2.45 x 10(-10), per-allele OR 0.77, 95% CI 0.71-0.84). A single SNP, rs401681 (P = 3.66 x 10(-7), per-allele OR 1.19, 95% CI 1.11-1.27), maps to the CLPTM1L-TERT locus on 5p15.33, which is associated with multiple cancers. Our study has identified common susceptibility loci for pancreatic cancer that warrant follow-up studies.


Long-term exposure to combination antiretroviral therapy and risk of death from specific causes: no evidence for any previously unidentified increased risk due to antiretroviral therapy.
Despite the known substantial benefits of combination antiretroviral therapy (cART), cumulative adverse effects could still limit the overall long-term treatment benefit. Therefore we investigated changes in the rate of death with increasing exposure to cART. A total of 12 069 patients were followed from baseline, which was defined as the time of starting cART or enrolment into EuroSIDA whichever occurred later, until death or 6 months after last follow-up visit. Incidence rates of death were calculated per 1000 person-years of follow-up (PYFU) and stratified by time of exposure to cART (≥3 antiretrovirals): less than 2, 2-3.99, 4-5.99, 6-7.99 and more than 8 years. Duration of cART exposure was the cumulative time actually receiving cART. Poisson regression models were fitted for each cause of death separately. A total of 1297 patients died during 70,613 PYFU [incidence rate 18.3 per 1000 PYFU, 95% confidence interval (CI) 17.4-19.4], 413 due to AIDS (5.85, 95% CI 5.28-6.41) and 884 due to non-AIDS-related cause (12.5, 95% CI 11.7-13.3). After adjustment for confounding variables, including baseline CD4 cell count and HIV RNA, there was a significant decrease in the rate of all-cause and AIDS-related death between 2 and 3.99 years and longer exposure time. In the first 2 years on cART the risk of non-AIDS death was significantly lower, but no significant difference in the rate of non-AIDS-related deaths between 2 and 3.99 years and longer exposure to cART was observed. In conclusion, we found no evidence of an increased risk of both all-cause and non-AIDS-related deaths with long-term cumulative cART exposure.


The value of coordinated services with court-referred clients and their families: an outcome study.
This study assessed the effectiveness of building partnerships with community resources and systems for court-referred clients and their families through a participant outcome evaluation. Specific variables studied included change in substance abuse patterns, family well-being, child safety, and recidivism. Results from pre-post testing revealed that a model with a single case coordinator who collaborated across service providers was particularly effective with court-referred clients and their families for increasing family intimacy and child well-being and for decreasing family danger and conflict. Discussion and recommendations are included.


Emergent cardiac surgery during transcatheter aortic valve implantation (TAVI): insights from the Edwards SAPIEN Aortic Bioprosthesis European Outcome (SOURCE) registry.
Transcatheter aortic valve implantation (TAVI) carries the risk of intraprocedural complications that may ultimately require emergent cardiac surgery (ECS). However, few data exist on the incidence, reasons and outcomes of patients needing ECS during TAVI. We analysed data from 2,307 TAVI patients, prospectively enrolled in the multicentre Edwards SAPIEN Aortic Bioprosthesis European Outcome (SOURCE) registry. Twenty-seven (1.2%) of 2,307 patients required ECS. The rates of ECS were similar for patients undergoing transapical TAVI compared with transfemoral TAVI (1.1% vs. 1.2%). The leading causes for ECS were embolisation/migration of the TAVI valve prosthesis (9/27, 33%) and procedure-related aortic injury (n=7, 26%). Thirty-day mortality of ECS was high (51.9%) and showed cause-specific differences, with 100% mortality in patients with aortic rupture or cardiac tamponade, 0% death in those with acute aortic regurgitation and intermediate risk of death or intermediate mortality in those with aortic injury or valve embolisation/migration. Rates of ECS during TAVI were low (1.2%). Although ECS was performed without time delay, emergent surgery was associated with a 30-day mortality of 52%. Complications with dramatic acute consequences (annular rupture, aortic injury) had higher mortality than those with less acute deterioration (aortic regurgitation). Prevention of complications requiring ECS during TAVI appears to be of critical importance, focusing on less traumatic, more flexible delivery catheter systems and retrievable valves to reduce the risk of aortic injury and valve embolisation, the two most common causes of ECS.


Confocal visualization of the effect of short-term locomotor exercise on BDNF and TrkB distribution in the lumbar spinal cord of the rat: the enhancement of BDNF in dendrites?
Locomotor exercise increases neurotrophin BDNF and its receptor TrkBFL expression in the lumbar spinal cord. Involvement of BDNF/TrkBFL in synaptic transmission raises the questions which intracellular compartments are involved in this upregulation and whether exercise leads to redistribution of these proteins related to the duration of exercise. We have investigated the influence of short-term (7 days) locomotor exercise (ST) on intracellular distribution of BDNF and TrkBFL in the rat lumbar spinal cord comparing it with the effects of long-term (28 days) exercise (LT) described earlier. Immunofluorescence (IF) of proteins was analyzed with confocal microscopy. ST exercise caused a redistribution of perikaryonal BDNF IF toward periphery resulting in an increase of dendritic signal. In contrast to an enhancement of perikaryonal BDNF staining following LT, no increase of BDNF IF in cell bodies was observed after ST. An increase of TrkBFL IF in oligodendrocytes was consistent with that caused by LT. The fibers of TrkBFL IF oligodendrocytes surrounding the largest neurons were in close apposition to neuronal membrane. We propose that ST exercise causes (1) BDNF translocation to dendrites and/or local dendritic synthesis to serve increased synaptic activity (2) sensitization of oligodendroglia to BDNF mediated responses.


Contact lens cases: the missing link in contact lens safety?
To summarize a variety of issues associated with contact lens case contamination and discuss appropriate methods that can limit this. A literature review was undertaken investigating the major factors associated with case contamination, with specific reference to the major pathogens associated with contamination, the role of bacterial biofilms, and methods that can limit contamination. The use of antimicrobial cases, regular case cleaning and case replacement, avoidance of topping up solutions, and not using tap water to rinse cases all appear to be important in avoidance of significant case contamination. Contact lens case contamination is a significant public health concern and may contribute significantly to the development of microbial keratitis in contact lens wearers. Patients should be reminded that they must clean and disinfect their lens cases daily, should avoid the use of tap water for rinsing them, must not top up their solutions, must take into careful consideration where and how the cases are stored during the time that lenses are being worn and that they must be replaced regularly. The adoption of these methods will substantially reduce the levels of contamination of cases with pathogenic microbes.


Changes in physical functioning in the Active Living Every Day program of the Active for Life Initiative®.
Physical activity can prevent or delay the onset of physical functional limitations in older adults. There are limited data that evidence-based physical activity interventions can be successfully translated into community programs and result in similar benefits for physical functioning. The purpose of this study is to measure the effects of the Active Living Every Day program on physical functioning and physical functional limitations in a diverse sample of older adults. As a part of the Active for Life initiative, the Council on Aging of Southwestern Ohio implemented Active Living Every Day (ALED), a group-based lifestyle behavior change program designed to increase physical activity. Performance-based physical functioning tests (30-s Chair Stand Test, eight Foot Up-and-Go Test, Chair Sit-and-Reach Test, 30-Foot Walk Test) were administered to participants at baseline and posttest. Baseline to post-program changes in physical functioning and impairment status were examined with repeated measures analysis of covariance. Interactions tested whether change over time differed according to race/ethnicity, body mass index (BMI), and baseline impairment status. Participants significantly increased their performance in all four physical functioning tests. The percentage of participants classified as "impaired" according to normative data significantly decreased over time. Physical functioning improved regardless of BMI, race/ethnicity, or baseline impairment status. ALED is an example of an evidenced-based physical activity program that can be successfully translated into community programs and result in significant and clinically meaningful improvements in performance-based measures of physical functioning.


Percutaneous cannulation for extracorporeal membrane oxygenation by intensivists: a retrospective single-institution case series.
Extracorporeal membrane oxygenation provides support for patients with severe acute cardiopulmonary failure, allowing the application of lung or myocardial rest in anticipation of organ recovery, or as a bridge to long-term support. Advances in technology have improved the safety and ease of application of extracorporeal membrane oxygenation. Percutaneous cannulation is one of these advances and is now preferred over surgical cannulation in most cases. Percutaneous cannulation is increasingly performed by intensivists, cardiologists, interventional radiologists, and related specialties. The objective of this study is to review the experience of percutaneous cannulation by intensivists at a single institution. A retrospective review of 100 subjects undergoing percutaneous cannulation for extracorporeal membrane oxygenation. Adult ICUs and PICUs at a tertiary academic medical institution. Critically ill neonatal, pediatric, and adult subjects with severe respiratory and/or cardiac failure undergoing percutaneous cannulation for extracorporeal membrane oxygenation. Modes of support included venoarterial, venovenous, venovenoarterial, and arteriovenous. Percutaneous extracorporeal membrane oxygenation. Case reports submitted to the Extracorporeal Life Support Organization and hospital records of the subjects were retrospectively reviewed. Subject demographics, type of support, cannulation configuration, types of cannulas, use of imaging modalities, and complications were recorded and summarized. One hundred ninety cannulations with cannula sizes from size 12 to 31F were performed by four intensivists in 100 subjects. Twenty-three were arterial (12-16F) and 167 were venous (12-31F). Preinsertion ultrasound was performed in 93 subjects (93%), fluoroscopic guidance in 79 subjects (85% of nonarteriovenous subjects), and ultrasound-guided insertion was performed in 65 subjects (65%). Two major complications occurred, each associated with mortality. Cannulation was successful in all other subjects (98% of subjects and 99% of cannulations). There were no cases of cannula-related bloodstream infection. Percutaneous cannulation for extracorporeal membrane oxygenation by intensivists can be performed with a high rate of success and a low rate of complications when accompanied by imaging support.


Glucosinolates in mixed-packaged mini broccoli and mini cauliflower under modified atmosphere.
Contents of total and individual glucosinolates of mini broccoli cv. Milady and mini cauliflower cv. Clarke were assessed to determine the effect of modified atmosphere packaging on postharvest glucosinolate dynamics of mixed mini Brassica vegetables. Therefore, mixed-packaged mini broccoli and mini cauliflower stored in food trays sealed with two different microperforated biaxial-oriented polypropylene films for up to 7 days at 8 degrees C were analyzed. The results indicate that modified atmosphere at 8% O2 + 14% CO2 was a suitable gaseous combination to maintain aliphatic and indole glucosinolates in mini broccoli for 7 days after an initial decrease at 4 days. In contrast, modified atmosphere at 1% O2 + 21% CO2 resulted in the best retention of indole glucosinolates of mini cauliflower for 7 days and also of aliphatic glucosinolates after an initial decrease at 4 days. Thus, to maintain glucosinolates and external appearance and to prevent off-odor, mini broccoli and mini cauliflower should be packed separately in suitable altered gas composition.


PINK1/Parkin-mediated mitophagy is dependent on VDAC1 and p62/SQSTM1.
Parkinson's disease is the most common neurodegenerative movement disorder. Mutations in PINK1 and PARKIN are the most frequent causes of recessive Parkinson's disease. However, their molecular contribution to pathogenesis remains unclear. Here, we reveal important mechanistic steps of a PINK1/Parkin-directed pathway linking mitochondrial damage, ubiquitylation and autophagy in non-neuronal and neuronal cells. PINK1 kinase activity and its mitochondrial localization sequence are prerequisites to induce translocation of the E3 ligase Parkin to depolarized mitochondria. Subsequently, Parkin mediates the formation of two distinct poly-ubiquitin chains, linked through Lys 63 and Lys 27. In addition, the autophagic adaptor p62/SQSTM1 is recruited to mitochondrial clusters and is essential for the clearance of mitochondria. Strikingly, we identified VDAC1 (voltage-dependent anion channel 1) as a target for Parkin-mediated Lys 27 poly-ubiquitylation and mitophagy. Moreover, pathogenic Parkin mutations interfere with distinct steps of mitochondrial translocation, ubiquitylation and/or final clearance through mitophagy. Thus, our data provide functional links between PINK1, Parkin and the selective autophagy of mitochondria, which is implicated in the pathogenesis of Parkinson's disease.


DNase I inhibits a late phase of reactive oxygen species production in neutrophils.
Neutrophils kill bacteria on extracellular complexes of DNA fibers and bactericidal proteins known as neutrophil extracellular traps (NETs). The NET composition and the bactericidal mechanisms they use are not fully understood. Here, we show that treatment with deoxyribonuclease (DNase I) impairs a late oxidative response elicited by Gram-positive and Gram-negative bacteria and also by phorbol ester. Isoluminol-dependent chemiluminescence elicited by opsonized Listeria monocytogenes-stimulated neutrophils was inhibited by DNase I, and the DNase inhibitory effect was also evident when phagocytosis was blocked, suggesting that DNase inhibits an extracellular mechanism of reactive oxygen species (ROS) generation. The DNase inhibitory effect was independent of actin polymerization. Phagocytosis and cell viability were not impaired by DNase I. Immunofluorescence analysis shows that myeloperoxidase is present on NETs. Furthermore, granular proteins were detected in NETs from Rab27a-deficient neutrophils which have deficient exocytosis, suggesting that exocytosis and granular protein distribution on NETs proceed by independent mechanisms. NADPH oxidase subunits were also detected on NETs, and the detection of extracellular trap-associated NADPH oxidase subunits was abolished by treatment with DNase I and dependent on cell stimulation. In vitro analyses demonstrate that MPO and NADPH oxidase activity are not directly inhibited by DNase I, suggesting that its effect on ROS production depends on NET disassembly. Altogether, our data suggest that inhibition of ROS production by microorganism-derived DNase would contribute to their ability to evade killing.


Music alters constitutively expressed opiate and cytokine processes in listeners.
Listening to music as a means of inducing a sense of calm and relaxation has been known for some time. Although these effects are robust, little research has been conducted into exploring the underlying neurochemical changes, which must occur to provide an individual with these objective sensations. In the current report we utilize an AB design to explore observed differences in blood plasma signaling molecules in pre- and post music listening groups when compared with controls. We focus chiefly on cytokines, as well as peripheral blood mononuclear cells and polymorphonuclear cells. We utilized reverse transcription followed by real-time polymerase chain reactions to determine relative mRNA expression for the mu opiate receptor gene. Using high pressure liquid chromatography coupled to electrochemical detection as well as nano electrospray ionization double quadrupole orthogonal acceleration time of flight-mass spectrometry we determined opiate alkaloid levels. Our findings are two-fold: with regard to mu opiate receptor expression, mononuclear cells showed a statistically significant increase in subjects in the music group compared to the control. Plasma morphine levels were found to be non-significantly lower in subjects after listening to music when compared to control subjects whereas morphine 6 glucuronide levels increased slightly, suggesting morphine's conversion to morphine 6 glucuronide. IL- 6 levels were significantly lower as well whereas IL-1b, IL-10 and cortisol values were unchanged. Taken together, it appears that music-listeners exhibit plasma signal molecule changes consistent with the physiological changes associated with the reported actions of music, i.e, lower blood pressure.


Oral administration of both tetrahydrobiopterin and L-arginine prevents endothelial dysfunction in rats with chronic renal failure.
We examined the mechanism of endothelial dysfunction in chronic renal failure (CRF), with reference to NO synthase. CRF was induced by 5/6 nephrectomy in rats. Either L-arginine (1.25 g/L in drinking water), tetrahydrobiopterin (BH4, 10 mg/kg per day in food), or a combination of the 2 were orally administered to CRF rats for 9 weeks. CRF rats showed elevation of systolic blood pressure compared with sham-operated rats. Endothelium-dependent relaxation induced by acetylcholine or A23187 in the isolated aorta was significantly reduced, and in vitro treatment with L-arginine, BH4, or superoxide dismutase restored the relaxation. Aortic segments from CRF rats showed significantly higher superoxide production in response to A23187, which was inhibited by L-NAME. Plasma concentrations of asymmetric dimethylarginine and symmetric dimethylarginine were higher in CRF rats. These changes in CRF rats were totally or partially decreased by L-arginine or BH4 supplementation in vivo. Interestingly, the combined treatment showed additive effects in certain parameters. These results suggest that vascular disorders in CRF rats may be partly due to NOS uncoupling caused by a relative deficiency of BH4 and partially due to accumulation of endogenous inhibitors of NOS and L-arginine uptake, resulting in the decrease of NO production and the increase of reactive oxygen species.


Connexin46fs380 causes progressive cataracts.
Although many connexin46 (Cx46) mutants have been linked to inherited human cataracts, there are no adequate animal models for their study. The current experiments were designed to characterize the consequences of expression of one such mutant, Cx46fs380, in the mouse lens. Mice expressing Cx46fs380 were generated by a knockin strategy. Levels and distribution of specific proteins were analyzed by immunoblotting and immunofluorescence. Dark-field microscopy revealed that lenses of young heterozygous and homozygous Cx46fs380 mice did not have opacities, but they developed anterior nuclear cataracts that became more severe with age. Immunofluorescence and immunoblotting showed that Cx46 was severely reduced in both heterozygous and homozygous Cx46fs380 lenses at 1 month of age, whereas immunoreactive connexin50 (Cx50) was moderately decreased. The reduction in Cx50 became more severe in older lenses. The solubilities of crystallins from young wild-type and fs380 mice were similar, but older fs380 lenses exhibited abnormalities of abundance, solubility, and modification of some crystallins. Major decreases in connexin levels precede the development of cataracts. These mice represent a useful model for elucidation of the progression of lens abnormalities during cataractogenesis especially as caused by a mutant connexin.


A rare missense variant in RCL1 segregates with depression in extended families.
Depression is the most prevalent psychiatric disorder with a complex and elusive etiology that is moderately heritable. Identification of genes would greatly facilitate the elucidation of the biological mechanisms underlying depression, however, its complex etiology has proved to be a major bottleneck in the identification of its genetic risk factors, especially in genome-wide association-like studies. In this study, we exploit the properties of a genetic isolate and its family-based structure to explore whether relatively rare exonic variants influence the burden of depressive symptoms in families. Using a multistep approach involving linkage and haplotype analyses followed by exome sequencing in the Erasmus Rucphen Family (ERF) study, we identified a rare (minor allele frequency (MAF)=1%) missense c.1114C>T mutation (rs115482041) in the RCL1 gene segregating with depression across multiple generations. Rs115482041 showed significant association with depressive symptoms (N=2393, βT-allele=2.33, P-value=1 × 10-4) and explained 2.9% of the estimated genetic variance of depressive symptoms (22%) in ERF. Despite being twice as rare (MAF<0.5%), c.1114C>T showed similar effect and significant association with depressive symptoms in samples from the independent population-based Rotterdam study (N=1604, βT-allele=3.60, P-value=3 × 10-2). A comparison of RCL1 expression in human and mouse brain revealed a striking co-localization of RCL1 with the layer 1 interlaminar subclass of astrocytes found exclusively in higher-order primates. Our findings identify RCL1 as a novel candidate gene for depression and offer insights into mechanisms through which RCL1 may be relevant for depression.


Walking or bicycling to school and weight status among adolescents from Montería, Colombia.
Active school transport (AST) is a recommended strategy to promote physical activity (PA) and prevent overweight (OW) in school-aged children. In many developing countries, such as Colombia, this association has not been well characterized. To determine the association between AST and weight status in a representative sample of adolescents from Montería, Colombia. Participants were 546 adolescents (278 boys) aged 11 to 18 years old from 14 randomly selected schools in Montería, Colombia in 2008. The PA module of the Global School Health Survey (GSHS-2007) was used to determine the prevalence of AST. To identify OW, participants were classified according to CDC 2000 criteria (BMI ≥ 85th percentile). Association between AST and OW was determined by binomial logistic regression. Odds ratios adjusted for age, sex, location of school, compliance with PA, and screen time recommendations showed that adolescents who reported AST had a significantly lower likelihood to be OW compared with adolescents who reported nonactive transportation (OR = 0.5, 95% CI 0.3-0.8, P < .05). These results support the importance of AST as a useful PA domain with potential implications for overweight prevention, in rapidly developing settings. Further epidemiologic and intervention studies addressing AST are needed in the region.


Recognition by association: Within- and cross-modality associative priming with faces and voices.
Recent literature has raised the suggestion that voice recognition runs in parallel to face recognition. As a result, a prediction can be made that voices should prime faces and faces should prime voices. A traditional associative priming paradigm was used in two studies to explore within-modality priming and cross-modality priming. In the within-modality condition where both prime and target were faces, analysis indicated the expected associative priming effect: The familiarity decision to the second target celebrity was made more quickly if preceded by a semantically related prime celebrity, than if preceded by an unrelated prime celebrity. In the cross-modality condition, where a voice prime preceded a face target, analysis indicated no associative priming when a 3-s stimulus onset asynchrony (SOA) was used. However, when a relatively longer SOA was used, providing time for robust recognition of the prime, significant cross-modality priming emerged. These data are explored within the context of a unified account of face and voice recognition, which recognizes weaker voice processing than face processing.


Estimating the contribution of assembly activity to cortical dynamics from spike and population measures.
The hypothesis that cortical networks employ the coordinated activity of groups of neurons, termed assemblies, to process information is debated. Results from multiple single-unit recordings are not conclusive because of the dramatic undersampling of the system. However, the local field potential (LFP) is a mesoscopic signal reflecting synchronized network activity. This raises the question whether the LFP can be employed to overcome the problem of undersampling. In a recent study in the motor cortex of the awake behaving monkey based on the locking of coincidences to the LFP we determined a lower bound for the fraction of spike coincidences originating from assembly activation. This quantity together with the locking of single spikes leads to a lower bound for the fraction of spikes originating from any assembly activity. Here we derive a statistical method to estimate the fraction of spike synchrony caused by assemblies-not its lower bound-from the spike data alone. A joint spike and LFP surrogate data model demonstrates consistency of results and the sensitivity of the method. Combining spike and LFP signals, we obtain an estimate of the fraction of spikes resulting from assemblies in the experimental data.


Adaptation of the growth hormone and insulin-like growth factor-I axis to chronic and severe calorie or protein malnutrition.
The hierarchy of diet components (e.g., protein, carbohydrate, vitamins, and minerals) influencing growth hormone (GH), insulin-like growth factor-I (IGF-I), and their binding proteins (BP) is not well defined. Young adult rats were fed diets for 1 mo that included low protein or 60% and 40% of carbohydrate calories. We hypothesized that levels of both hormones, their dominant BPs and liver IGF-I mRNA would fall, and that part of the mechanism for decreasing serum IGF-I would be enhanced IGFBP-3 protease activity. By day 30, caloric deprivation to 40% lowered serum GH, GHBP, IGF-I and IGFBP-3, and liver IGF-I mRNA. This was the only condition resulting in body weight loss (-15%) vs 39% gain in controls. Restriction to 60% calories had no impact on BP levels, slightly lowered IGF-I (-12%) in the face of a 95% inhibition of GH levels, while allowing a modest 9% body weight gain. Protein deprivation lowered serum GH, IGF-I and IGFBP-3, and liver IGF-I mRNA, while GHBP levels were normal. The reduced total IGF-I under these dietary conditions could not be explained by an increase in IGFBP-3 protease activity, or a decrease in the association of IGF-I with IGFBP-3 and the acid labile subunit.


Virulence evolution in response to vaccination: the case of malaria.
One theory of why some pathogens are virulent (i.e., they damage their host) is that they need to extract resources from their host in order to compete for transmission to new hosts, and this resource extraction can damage the host. Here we describe our studies in malaria that test and support this idea. We go on to show that host immunity can exacerbate selection for virulence and therefore that vaccines that reduce pathogen replication may select for more virulent pathogens, eroding the benefits of vaccination and putting the unvaccinated at greater risk. We suggest that in disease contexts where wild-type parasites can be transmitted through vaccinated hosts, evolutionary outcomes need to be considered.


Genome-wide association study identifies five new schizophrenia loci.
We examined the role of common genetic variation in schizophrenia in a genome-wide association study of substantial size: a stage 1 discovery sample of 21,856 individuals of European ancestry and a stage 2 replication sample of 29,839 independent subjects. The combined stage 1 and 2 analysis yielded genome-wide significant associations with schizophrenia for seven loci, five of which are new (1p21.3, 2q32.3, 8p23.2, 8q21.3 and 10q24.32-q24.33) and two of which have been previously implicated (6p21.32-p22.1 and 18q21.2). The strongest new finding (P = 1.6 × 10(-11)) was with rs1625579 within an intron of a putative primary transcript for MIR137 (microRNA 137), a known regulator of neuronal development. Four other schizophrenia loci achieving genome-wide significance contain predicted targets of MIR137, suggesting MIR137-mediated dysregulation as a previously unknown etiologic mechanism in schizophrenia. In a joint analysis with a bipolar disorder sample (16,374 affected individuals and 14,044 controls), three loci reached genome-wide significance: CACNA1C (rs4765905, P = 7.0 × 10(-9)), ANK3 (rs10994359, P = 2.5 × 10(-8)) and the ITIH3-ITIH4 region (rs2239547, P = 7.8 × 10(-9)).


Protonation states of remote residues affect binding-release dynamics of the ligand but not the conformation of apo ferric binding protein.
We have studied the apo (Fe(3+) free) form of periplasmic ferric binding protein (FbpA) under different conditions and we have monitored the changes in the binding and release dynamics of H2PO4(-) that acts as a synergistic anion in the presence of Fe(3+). Our simulations predict a dissociation constant in agreement with the experimentally measured value under the same ionic pH conditions. We apply perturbations relevant for changes in environmental conditions as (i) different values of ionic strength and (ii) protonation of a group of residues to mimic a different pH environment. Local perturbations are also studied by protonation or mutation of a site distal to the binding region that is known to mechanically manipulate the hinge-like motions of FbpA. We find that while the average conformation of the protein is intact in all simulations, the H2PO4(-) binding constant is substantially altered by the changing conditions. In particular, the bound fraction which is 20% for the wild type system is increased to 50% with a D52A mutation/protonation and further to over 90% at the protonation conditions mimicking those at pH 5.5. The change in the dynamics is traced to the altered electrostatic potential distribution on the surface of the protein which in turn affects hydrogen bonding patterns at the active site. The observations are also quantified by rigorous free energy calculations. Our results lend clues as to how the environment versus single residue perturbations may be utilized for regulation of binding modes in hFbpA systems in the absence of conformational changes.


Omega-6: Omega-3 PUFA Ratio, Pain, Functioning, and Distress in Adults With Knee Pain.
Osteoarthritis (OA) is associated with inflammation, chronic pain, functional limitations, and psychosocial distress. High omega-3 (n-3) polyunsaturated fatty acids (PUFAs) are associated with lower levels of inflammatory mediators, anti-nociception, and adaptive cognitive/emotional functioning. High omega-6 (n-6) PUFAs are associated with inflammation, nociception, and psychological distress. While findings related to n-3 supplementation in knee OA are mixed, consideration of the n-6:n-3 ratio and additional outcome measures may provide improved understanding of the potential relevance of these fatty acids in OA. On the basis of recommended and typical ranges of the n-6:n-3 ratio, we hypothesized that in adults with knee pain, those with a high n-6:n-3 ratio would have greater pain/functional limitations, experimental pain sensitivity, and psychosocial distress compared with those with a low n-6:n-3 ratio. A cross-sectional investigation of clinical and experimental pain and physical and psychosocial functioning was completed in 167 adults ages 45 to 85 meeting knee OA screening criteria. Blood samples were collected and the plasma n-6:n-3 PUFA ratio determined. Quartile splits were computed and low (n=42) and high (n=41) ratio groups were compared. The high ratio group reported greater pain and functional limitations, (all Ps<0.04), mechanical temporal summation (hand and knee, P<0.05), and perceived stress (P=0.008) but not depressive symptoms. In adults with knee pain, a high n-6:n-3 ratio is associated with greater clinical pain/functional limitations, experimental pain sensitivity, and psychosocial distress compared with a low ratio group. Findings support consideration of the n-6:n-3 PUFA ratio and additional clinical endpoints in future research efforts.


Biology of platelet-rich plasma and its clinical application in cartilage repair.
Platelet-rich plasma (PRP) is an autologous concentrated cocktail of growth factors and inflammatory mediators, and has been considered to be potentially effective for cartilage repair. In addition, the fibrinogen in PRP may be activated to form a fibrin matrix to fill cartilage lesions, fulfilling the initial requirements of physiological wound healing. The anabolic, anti-inflammatory and scaffolding effects of PRP based on laboratory investigations, animal studies, and clinical trials are reviewed here. In vitro, PRP is found to stimulate cell proliferation and cartilaginous matrix production by chondrocytes and adult mesenchymal stem cells (MSCs), enhance matrix secretion by synoviocytes, mitigate IL-1β-induced inflammation, and provide a favorable substrate for MSCs. In preclinical studies, PRP has been used either as a gel to fill cartilage defects with variable results, or to slow the progression of arthritis in animal models with positive outcomes. Findings from current clinical trials suggest that PRP may have the potential to fill cartilage defects to enhance cartilage repair, attenuate symptoms of osteoarthritis and improve joint function, with an acceptable safety profile. Although current evidence appears to favor PRP over hyaluronan for the treatment of osteoarthritis, the efficacy of PRP therapy remains unpredictable owing to the highly heterogeneous nature of reported studies and the variable composition of the PRP preparations. Future studies are critical to elucidate the functional activity of individual PRP components in modulating specific pathogenic mechanisms.


microRNAs and cancer metabolism reprogramming: the paradigm of metformin.
Increasing evidence witnesses that cancer metabolism alterations represent a critical hallmark for many types of human tumors. There is a strong need to understand and dissect the molecular mechanisms underlying cancer metabolism to envisage specific biomarkers and underpin critical molecular components that might represent novel therapeutic targets. One challenge, that is the focus of this review, is the reprogramming of the altered metabolism of a cancer cell toward that of un-transformed cell. The anti-hyperglicemic agent, metformin has proven to be effective in reprogramming the metabolism of cancer cells even from those subpopulations endowed with cancer stem like features and very high chemoresistenace to conventional anticancer treatments. A functional interplay involving selective modulation of microRNAs (miRNAs) takes place along the anticancer metabolic effects exerted by metformin. The implications of this interplay will be also discussed in this review.


Eight-year trends of cardiometabolic morbidity and mortality in patients with schizophrenia.
We examined cardiometabolic disease and mortality over 8 years among individuals with and without schizophrenia. We compared 65,362 patients in the Veteran Affairs (VA) health system with schizophrenia to 65,362 VA patients without serious mental illness (non-SMI) matched on age, service access year and location. The annual prevalence of diagnosed cardiovascular disease, diabetes, dyslipidemia, hypertension, obesity, and all-cause and cause-specific mortality was compared for fiscal years 2000-2007. Mean years of potential life lost (YPLLs) were calculated annually. The cohort was mostly male (88%) with a mean age of 54 years. Cardiometabolic disease prevalence increased in both groups, with non-SMI patients having higher disease prevalence in most years. Annual between-group differences ranged from <1% to 6%. Annual mortality was stable over time for schizophrenia (3.1%) and non-SMI patients (2.6%). Annual mean YPLLs increased from 12.8 to 15.4 in schizophrenia and from 11.8 to 14.0 for non-SMI groups. VA patients with and without schizophrenia show increasing but similar prevalence rates of cardiometabolic diseases. YPLLs were high in both groups and only slightly higher among patients with schizophrenia. The findings highlight the complex population served by the VA while suggesting a smaller mortality impact from schizophrenia than previously reported.


Antimicrobial resistance of Salmonella serotypes isolated from slaughter-age pigs and environmental samples.
The aim of this study was to determine the antimicrobial resistance patterns of Salmonella strains isolated from slaughter-age pigs and environmental samples collected at modern swine raising facilities in Brazil. Seventeen isolates of six serotypes of Salmonella enterica subsp. enterica were isolated out of 1,026 collected samples: Salmonella Typhimurium (1), Salmonella Agona (5), Salmonella Sandiego (5), Salmonella Rissen (1), Salmonella Senftenberg (4), and Salmonella Javiana (1). Resistance patterns were determined to extended-spectrum penicillin (ampicillin), broad-spectrum cephalosporins (cefotaxime and ceftriaxone), aminoglycosides (streptomycin, neomycin, gentamicin, amikacin, and tobramycin), narrow-spectrum quinolone (nalidixic acid), broad-spectrum quinolone (ciprofloxacin and norfloxacin), tetracycline, trimethoprim, and chloramphenicol. Antimicrobial resistance patterns varied among serotypes, but isolates from a single serotype consistently showed the same resistance profile. All isolates were resistant to tetracycline, streptomycin, and nalidixic acid. One isolate, Salmonella Rissen, was also resistant to cefotaxime and tobramycin. All serotypes were susceptible to ceftriaxone, norfloxacin, ciprofloxacin, ampicillin, gentamicin, and chloramphenicol. The high resistance to tetracycline and streptomycin may be linked to their common use as therapeutic drugs on the tested farms. No relation was seen between nalidixic acid and fluoroquinolone resistance.


Correlation of the plasma sphingoid base profile with results from oral glucose tolerance tests in gestational diabetes mellitus.
Oral glucose tolerance test (OGTT) is usually insufficient to accurately predict the risk for type 2 diabetes mellitus (T2DM), it is therefore necessary to identify an additional biomarker that would most likely improve the accuracy of OGTT. The current OGTT was performed in 53 volunteers after ingestion of 75 g glucose in 250 ml water to each volunteer. Similarly the sphingoid base profile of these volunteers was explored using liquid-chromatography linked with mass spectrometer (LC-MS) and correlated with the different time-points glucose values of OGTT as well as with total area under the curve (tAUC), incremental area under the curve (iAUC), and positive incremental area under the curve (pAUC). The findings showed that 1-deoxysphinganine (1-deoxySA) was significantly positively correlated with the 1-hour, 2-hour, and 3-hour plasma glucose level as well as with total, incremental, and positive incremental AUC while 1-deoxysphingosine (1-deoxySO) was correlated only with 1-hour, 2-hour glucose levels and tAUC of OGTT. The C18SAdiene was negatively correlated with all-time points glucose values and AUCs followed by negative correlation of C18SO, C16SO and C17SO with 2-hour glucose and tAUC of OGTT. The ratios of 1-deoxySA and 1-deoxySO with respect to C18SAdiene have shown significant correlation with 2-hour and AUCs. These ratios were higher in subjects with gestational diabetes in comparison with normal subjects. These findings underlined that 1-deoxysphingolipids (1-deoxySLs) and their ratios with C18SAdiene could be significantly correlated with the glucose load of OGTT and might be used as predictive biomarkers along with OGTT for the risk assessment of diabetes.


Group A rotaviruses in children with gastroenteritis in a Canadian pediatric hospital: The prevaccine era.
A publicly funded, group A rotavirus (RVA) vaccination program was implemented in Quebec in November 2011. To evaluate trends in RVA infections and describe circulating genotypes before the implementation of a publicly funded vaccination program. The Montreal Children's Hospital (Montreal, Quebec) virology laboratory database was reviewed for RVA ELISA performed between July 2006 and June 2011. A five-week moving average was used to follow the proportion of positive RVA ELISA test results. A season was defined as starting with the first two and ending with the final two consecutive weeks in which the percentage of specimens testing positive for RVA was ≥10%. Duplicate tests were excluded. A random sample of 39 RVA-positive fecal samples from the final season (2010/2011) was genetically characterized: VP4, VP6, VP7 and NSP4 gene segments were genotyped using sequence analysis. Of the 3403 nonduplicate tests, 433 were RVA positive: 15.1% (2006/2007) to 9.3% (2010/2011) of the samples were positive during the study period, with a proportionally larger decrease in the percentage of positive tests compared with the decrease in the number of tests performed. The most common RVA strain types detected were G9P[8]I1 (n=19) and G1P[8]I1 (n=14), followed by G2P[4]I2 (n=4), G3P[6]I1 (n=1) and G4P[8]I2 (n=1). Mixed RVA infection was observed in two samples. Before the implementation of the vaccination program, the proportion of positive RVA tests had already begun to steadily decline. The present study was the first to report the genetic makeup of human RVA collected from a Canadian hospital based on the genotyping of four gene segments. The present study provided a baseline with which to monitor the impact of the universal vaccination program.


Genomic biomarkers in prostate cancer.
Prostate cancer is the most common non-cutaneous cancer among men in the United States. In the last decade there has been a rapid expansion in the field of biomarker assays for diagnosis, prognosis, and treatment prediction in prostate cancer. The evidence base for these assays is rapidly evolving. With several commercial assays available at each stage of the disease, deciding which genomic assays are appropriate for which patients can be nuanced for physicians. In an effort to help guide these decisions in clinical practice, we aim to give an update on the current status of the biomarker field of prostate cancer.


Daily tea drinking is associated with a low level of depressive symptoms in the Finnish general population.
Tea drinking has been suggested to be beneficial in neurodegenerative diseases where depressive mood is a common symptom. Nevertheless, it is not known whether there are any associations between tea drinking and depression in general populations. In this study we investigated these associations in a sample of the Finnish general population (n = 2011) using a postal questionnaire and the Beck Depression Inventory (BDI). Those who reported drinking tea daily were less depressed than the others. They had a lower mean BDI score and also a lower prevalence of depression. None of those whose daily tea intake was five cups or more had depression. Several potential confounding factors were included in the final sex- and age-adjusted multivariate logistic regression model which suggested that those who drink tea daily may have a significantly reduced risk of being depressed (adjusted odds ratio 0.47, 95% confidence interval 0.27-0.83). In conclusion, an inverse relationship between daily tea drinking and the risk of being depressed was found in a relatively large general population sample. Nevertheless, the underlying mechanisms are unresolved and further studies are needed.


Decoding the androgen receptor splice variants.
In the past five years, multiple structurally and functionally distinct androgen receptor (AR) splice variants have been decoded and characterized. The mature transcripts for the majority of the fully decoded AR splice variants contain a transcribed "intronic" sequence, capable of encoding a short variant-specific peptide to replace the AR ligand-binding domain (LBD). Functionally, AR splice variants represent a diverse group of molecules often demonstrating cell context-specific genomic functions that may or may not be coupled with the functions of the canonical full-length AR (AR-FL). However, the full spectrum of their functional diversity and the underlying mechanistic basis remains very poorly characterized. In clinical specimens derived from men treated with a variety of hormone therapy regimens, AR splice variants are almost always expressed at detectable, yet lower levels when compared to that of AR-FL. In spite of the collective in vitro data supporting the putative role of AR splice variants in therapeutic resistance to hormone therapies, the extent to which AR splice variants mediate resistance to each individual regimen is not known and awaits thorough investigations in a clinically relevant setting using specimens from men undergoing treatments. Among the AR splice variants, AR-V7 is more abundantly and frequently expressed in castration-resistant prostate cancer (CRPC) and remains the most important variant identified so far. The relative importance of different AR molecules, including AR-FL, should be functionally dissected in the setting of castration-resistant prostate cancer, particularly in tumors resistant to more potent inhibitors of AR-FL recently approved by the FDA. In this review, we will focus on the discovery and characterization of AR splice variants, their putative functions and roles in mediating constitutively active AR signaling, and key areas of investigation that are necessary to establish their clinical relevance.


Neural Signatures of Value Comparison in Human Cingulate Cortex during Decisions Requiring an Effort-Reward Trade-off.
Integrating costs and benefits is crucial for optimal decision-making. Although much is known about decisions that involve outcome-related costs (e.g., delay, risk), many of our choices are attached to actions and require an evaluation of the associated motor costs. Yet how the brain incorporates motor costs into choices remains largely unclear. We used human fMRI during choices involving monetary reward and physical effort to identify brain regions that serve as a choice comparator for effort-reward trade-offs. By independently varying both options' effort and reward levels, we were able to identify the neural signature of a comparator mechanism. A network involving supplementary motor area and the caudal portion of dorsal anterior cingulate cortex encoded the difference in reward (positively) and effort levels (negatively) between chosen and unchosen choice options. We next modeled effort-discounted subjective values using a novel behavioral model. This revealed that the same network of regions involving dorsal anterior cingulate cortex and supplementary motor area encoded the difference between the chosen and unchosen options' subjective values, and that activity was best described using a concave model of effort-discounting. In addition, this signal reflected how precisely value determined participants' choices. By contrast, separate signals in supplementary motor area and ventromedial prefrontal cortex correlated with participants' tendency to avoid effort and seek reward, respectively. This suggests that the critical neural signature of decision-making for choices involving motor costs is found in human cingulate cortex and not ventromedial prefrontal cortex as typically reported for outcome-based choice. Furthermore, distinct frontal circuits seem to drive behavior toward reward maximization and effort minimization. The neural processes that govern the trade-off between expected benefits and motor costs remain largely unknown. This is striking because energetic requirements play an integral role in our day-to-day choices and instrumental behavior, and a diminished willingness to exert effort is a characteristic feature of a range of neurological disorders. We use a new behavioral characterization of how humans trade off reward maximization with effort minimization to examine the neural signatures that underpin such choices, using BOLD MRI neuroimaging data. We find the critical neural signature of decision-making, a signal that reflects the comparison of value between choice options, in human cingulate cortex, whereas two distinct brain circuits drive behavior toward reward maximization or effort minimization.


Development of the Community Healthy Living Index: a tool to foster healthy environments for the prevention of obesity and chronic disease.
This paper presents a new, comprehensive tool for communities to assess opportunities for active living and healthy eating and to mobilize all sectors of society to conquer obesity and chronic disease. Relevant existing tools and input from an expert panel were considered to draft the Community Healthy Living Index (CHLI). CHLI covers five major sectors where people live, work, learn, and play: schools, afterschools, work sites, neighborhoods, and the community-at-large. CHLI and the accompanying procedures enable community teams to assess programs, the physical environment, and policies related to healthy living and to plan improvement strategies. In 2008, with local YMCAs acting as conveners, community assessment teams from six US communities pilot-tested CHLI for cognitive response testing, inter-rater reliability, and implementation feasibility. CHLI was revised to reflect the test results. Pilot analyses demonstrated that the process was feasible, with most questions being interpreted as intended and showing substantial to almost perfect agreement between raters. The final CHLI is being disseminated nationally. Preliminary data illustrate CHLI obtains reliable results and is feasible to implement. CHLI is a promising tool for community-based prevention efforts to draw attention to opportunities for healthy living and create impetus for community changes.