Increasing User Involvement in Health Care and Health Research Simultaneously: A Proto-Protocol for "Person-as-Researcher" and Online Decision Support Tools.
User involvement is appearing increasingly on policy agendas in many countries, with a variety of proposals for facilitating it. The belief is that it will produce better health for individuals and community, as well as demonstrate greater respect for the basic principles of autonomy and democracy. Our Web-based project aims to increase involvement in health care and health research and is presented in the form of an umbrella protocol for a set of project-specific protocols. We conceptualize the person as a researcher engaged in a continual, living, informal "n-of-1"-type study of the effects of different actions and interventions on their health, including those implying contact with health care services. We see their research as primarily carried out in order to make better decisions for themselves, but they can offer to contribute the results to the wider population. We see the efforts of the "person-as-researcher" as contributing to the total amount of research undertaken in the community, with research not being confined to that undertaken by professional researchers and institutions. This view is fundamentally compatible with both the emancipatory and conventional approaches to increased user involvement, though somewhat more aligned with the former. Our online decision support tools, delivered directly to the person in the community and openly accessible, are to be seen as research resources. They will take the form of interactive decision aids for a variety of specific health conditions, as well as a generic one that supports all health and health care decisions through its focus on key aspects of decision quality. We present a high-level protocol for the condition-specific studies that will implement our approach, organized within the Populations, Interventions, Comparators, Outcomes, Timings, and Settings (PICOTS) framework. Our underlying hypothesis concerns the person-as-researcher who is equipped with a prescriptive, transparent, expected value-based opinion-an opinion that combines their criterion importance weights with the Best Estimates Available Now for how well each of the available options performs on each of those outcomes. The hypothesis is that this person-as-researcher is more likely to be able to position themselves as an active participant in a clinical encounter, if they wish, than someone who has engaged with a descriptive decision aid that attempts to work with their existing cognitive processes and stresses the importance of information. The precise way this is hypothesis tested will be setting-specific and condition-specific and will be spelled out in the individual project protocols. Decision resources that provide fast access to the results of slower thinking can provide the stimulus that many individuals need to take a more involved role in their own health. Our project, advanced simply as one approach to increased user involvement, is designed to make progress in the short term with minimal resources and to do so at the point of decision need, when motivation is highest. Some basic distinctions, such as those between science and non-science, research and practice, community and individual, and lay and professional become somewhat blurred and may need to be rethought in light of this approach.


PET/CT versus MRI for diagnosis, staging, and follow-up of lung cancer.
Positron emission tomography / computed tomography (PET/CT), with its metabolic data of (18) F-fluorodeoxyglucose (FDG) cellular uptake in addition to morphologic CT data, is an established technique for staging of lung cancer and has higher sensitivity and accuracy for lung nodule characterization than conventional approaches. Its strength extends outside the chest, with unknown metastases detected or suspected metastases excluded in a significant number of patients. Lastly, PET/CT is used in the assessment of therapy response. Magnetic resonance imaging (MRI) in the chest has been difficult to establish, but with the advent of new sequences is starting to become an increasingly useful alternative to conventional approaches. Diffusion-weighted MRI (DWI) is useful for distinguishing benign and malignant pulmonary nodules, has high sensitivity and specificity for nodal staging, and is helpful for evaluating an early response to systemic chemotherapy. Whole-body MRI/PET promises to contribute additional information with its higher soft-tissue contrast and much less radiation exposure than PET/CT and has become feasible for fast imaging and can be used for cancer staging in patients with a malignant condition.


Broad bean (Vicia faba L.) pods: a rich source of bioactive ingredients with antimicrobial, antioxidant, enzyme inhibitory, anti-diabetic and health-promoting properties.
This study was aimed at investigating the chemical composition (proximate, minerals, fatty acids and phenolic compounds) and the in vitro (antimicrobial, radical scavenging, anti-acetylcholinesterase and protein denaturing activities) and in vivo (anti-diabetic and histo-protective effects in alloxan-induced diabetic mice) biological activities of broad bean pods (BBPs), a food waste by-product material. The results showed that BBPs have high dietary fiber (57.46%), carbohydrate (18.93%) and protein (13.81%) content versus low fat content (<1%) contributing to a low energy value of 139.24 kcal per 100 g. Profiling of fatty acids showed an abundance of the essential polyunsaturated α-linolenic and linoleic acids, exhibiting an excellent nutritional quality as revealed by their low atherogenic and thrombogenic indices and their hypocholesterolemic properties. The methanol extract which exhibited the highest total phenolic, flavonoid and tannin contents was found to be the most active extract in terms of antimicrobial and anti-radical activities. In alloxan-induced diabetic mice, the oral administration of a methanol extract (500 mg per kg bw) attenuated the elevated levels of serum alanine aminotransferase (ALA), aspartate aminotransferase (AST), and alkaline phosphatase activities, and urea, uric acid, and creatinine. It effectively normalized the status of lipid profiles, mitigated oxidative stress through the activation of antioxidant enzymes (CAT, GPx and SOD), and alleviated oxidative stress-mediated histopathological changes in the pancreas, liver, kidney and testis. Compositional analysis by HPLC-PDA-ESI-MS/MS revealed the presence of flavan-3-ols (catechin, epicatechin and their derivatives), flavones (apigenin derivatives) and flavonols (glycosides of quercetin and kaempferol), among others. These findings suggest that BBPs may be an effective functional food for the management of diabetes and its complications.


Getting the message across: mechanisms of physiological cross talk by adipose tissue.
Obesity is associated with resistance of skeletal muscle to insulin-mediated glucose uptake, as well as resistance of different organs and tissues to other metabolic and vascular actions of insulin. In addition, the body is exquisitely sensitive to nutrient imbalance, with energy excess or a high-fat diet rapidly increasing insulin resistance, even before noticeable changes occur in fat mass. There is a growing acceptance of the fact that, as well as acting as a storage site for surplus energy, adipose tissue is an important source of signals relevant to, inter alia, energy homeostasis, fertility, and bone turnover. It has also been widely recognized that obesity is a state of low-grade inflammation, with adipose tissue generating substantial quantities of proinflammatory molecules. At a cellular level, the understanding of the signaling pathways responsible for such alterations has been intensively investigated. What is less clear, however, is how alterations of physiology, and of signaling, within one cell or one tissue are communicated to other parts of the body. The concepts of cell signals being disseminated systemically through a circulating "endocrine" signal have been complemented by the view that local signaling may similarly occur through autocrine or paracrine mechanisms. Yet, while much elegant work has focused on the alterations in signaling that are found in obesity or energy excess, there has been less attention paid to ways in which such signals may propagate to remote organs. This review of the integrative physiology of obesity critically appraises the data and outlines a series of hypotheses as to how interorgan cross talk takes place. The hypotheses presented include the "fatty acid hypothesis,", the "portal hypothesis,", the "endocrine hypothesis,", the "inflammatory hypothesis,", the "overflow hypothesis,", a novel "vasocrine hypothesis," and a "neural hypothesis," and the strengths and weaknesses of each hypothesis are discussed.


Gender hormones and the progression of experimental polycystic kidney disease.
Male gender is a risk factor for progression of autosomal-dominant polycystic kidney disease (ADPKD), clinically and in the Han:SPRD rat model. Orchiectomy limits progression, but mechanisms of the detrimental effect of androgen, and/or beneficial effects of estrogen, are not known. This protocol tested the hypothesis that male gender (intact androgen status) promotes progression, while female gender (intact estrogen status) is protective; and that these disease-modifying effects are due to changes in expression of known fibrotic mediators. Studies were performed in male and female noncystic control (+/+) and cystic (+/-) rats subjected to orchiectomy, ovariectomy, or sham operation. At 12 weeks of age, renal function was measured. Blood and kidneys were taken for measurement of plasma and renal renin, endothelin (ET-1), endothelial nitric oxide synthase (eNOS), and vascular endothelial growth factor (VEGF), using biochemical, protein expression, and immunohistochemical methods. Cystic male rats exhibited significantly reduced glomerular filtration (GFR) and effective renal plasma flow (ERPF) rates, with suppression of plasma and renal renin, up-regulation of renal ET-1 and eNOS, and down-regulation of renal VEGF expression. Orchiectomy attenuated the fall in GFR and ERPF, while numerically limiting changes in eNOS and VEGF. Female rats exhibited less cystic growth, with normal renin status, lesser elevation of renal ET-1, and proportionately lesser changes in VEGF and eNOS. Ovariectomy led to higher blood pressure and reduced GFR and ERPF, with a trend toward upregulation of ET-1, and significant down-regulation of VEGF and eNOS. Female gender is protective, but ovariectomy attenuates the protective effect of female gender, in association with changes in renal expression of ET-1, VEGF, and eNOS. The accelerated disease in male rats can be attenuated by orchiectomy and consequent changes in expression of disease mediators.


Ultra-fast digital tomosynthesis reconstruction using general-purpose GPU programming for image-guided radiation therapy.
The purpose of this work is to demonstrate an ultra-fast reconstruction technique for digital tomosynthesis (DTS) imaging based on the algorithm proposed by Feldkamp, Davis, and Kress (FDK) using standard general-purpose graphics processing unit (GPGPU) programming interface. To this end, the FDK-based DTS algorithm was programmed "in-house" with C language with utilization of 1) GPU and 2) central processing unit (CPU) cards. The GPU card consisted of 480 processing cores (2 x 240 dual chip) with 1,242 MHz processing clock speed and 1,792 MB memory space. In terms of CPU hardware, we used 2.68 GHz clock speed, 12.0 GB DDR3 RAM, on a 64-bit OS. The performance of proposed algorithm was tested on twenty-five patient cases (5 lung, 5 liver, 10 prostate, and 5 head-and-neck) scanned either with a full-fan or half-fan mode on our cone-beam computed tomography (CBCT) system. For the full-fan scans, the projections from 157.5°-202.5° (45°-scan) were used to reconstruct coronal DTS slices, whereas for the half-fan scans, the projections from both 157.5°-202.5° and 337.5°-22.5° (2 x 45°-scan) were used to reconstruct larger FOV coronal DTS slices. For this study, we chose 45°-scan angle that contained ~80 projections for the full-fan and ~160 projections with 2 x 45°-scan angle for the half-fan mode, each with 1024 x 768 pixels with 32-bit precision. Absolute pixel value differences, profiles, and contrast-to-noise ratio (CNR) calculations were performed to compare and evaluate the images reconstructed using GPU- and CPU-based implementations. The time dependence on the reconstruction volume was also tested with (512 x 512) x 16, 32, 64, 128, and 256 slices. In the end, the GPU-based implementation achieved, at most, 1.3 and 2.5 seconds to complete full reconstruction of 512 x 512 x 256 volume, for the full-fan and half-fan modes, respectively. In turn, this meant that our implementation can process > 13 projections-per-second (pps) and > 18 pps for the full-fan and half-fan modes, respectively. Since commercial CBCT system nominally acquires 11 pps (with 1 gantry-revolution-per-minute), our GPU-based implementation is sufficient to handle the incoming projections data as they are acquired and reconstruct the entire volume immediately after completing the scan. In addition, on increasing the number of slices (hence volume) to be reconstructed from 16 to 256, only minimal increases in reconstruction time were observed for the GPU-based implementation where from 0.73 to 1.27 seconds and 1.42 to 2.47 seconds increase were observed for the full-fan and half-fan modes, respectively. This resulted in speed improvement of up to 87 times compared with the CPU-based implementation (for 256 slices case), with visually identical images and small pixel-value discrepancies (< 6.3%), and CNR differences (< 2.3%). With this achievement, we have shown that time allocation for DTS image reconstruction is virtually eliminated and that clinical implementation of this approach has become quite appealing. In addition, with the speed achievement, further image processing and real-time applications that was prohibited prior due to time restrictions can now be tempered with.


Reported sleep patterns in obsessive compulsive disorder (OCD).
Previous studies of sleep disturbance in OCD have produced contradictory findings. This may be due to different methodological approaches but may also reflect the fact that many studies have not screened OCD patients for depressive symptoms. This is particularly important since major depression is well known to disrupt sleep. The current study examined 94 subjective reports of sleep as measured by the Pittsburgh Sleep Quality Index (PSQI). Twenty-four OCD patients with and without co-morbid depression ( n =12 in each group) were compared with a group of normal subjects ( n =57) and a group of subjects with major depression ( n =13). In general, the OCD patients with co-morbid depression exhibited very similar sleep patterns to the group with major depression. By contrast, those OCD patients without depressive symptoms showed similar sleep patterns to the group of normal subjects. Thus, contrary to some earlier studies, these data suggest that OCD does not generally influence reported sleep patterns unless there is a co-morbid diagnosis. However, a few non-depressed OCD patients did report sleep phase shifting, suggesting that a small proportion may have abnormal sleep parameters. Such anomalies may be masked by group analyses and further case studies are therefore warranted to investigate whether such patients exhibit abnormal scores on biological sleep markers.


Correction of diverse muscular dystrophy mutations in human engineered heart muscle by single-site genome editing.
Genome editing with CRISPR/Cas9 is a promising new approach for correcting or mitigating disease-causing mutations. Duchenne muscular dystrophy (DMD) is associated with lethal degeneration of cardiac and skeletal muscle caused by more than 3000 different mutations in the X-linked dystrophin gene (DMD). Most of these mutations are clustered in "hotspots." There is a fortuitous correspondence between the eukaryotic splice acceptor and splice donor sequences and the protospacer adjacent motif sequences that govern prokaryotic CRISPR/Cas9 target gene recognition and cleavage. Taking advantage of this correspondence, we screened for optimal guide RNAs capable of introducing insertion/deletion (indel) mutations by nonhomologous end joining that abolish conserved RNA splice sites in 12 exons that potentially allow skipping of the most common mutant or out-of-frame DMD exons within or nearby mutational hotspots. We refer to the correction of DMD mutations by exon skipping as myoediting. In proof-of-concept studies, we performed myoediting in representative induced pluripotent stem cells from multiple patients with large deletions, point mutations, or duplications within the DMD gene and efficiently restored dystrophin protein expression in derivative cardiomyocytes. In three-dimensional engineered heart muscle (EHM), myoediting of DMD mutations restored dystrophin expression and the corresponding mechanical force of contraction. Correcting only a subset of cardiomyocytes (30 to 50%) was sufficient to rescue the mutant EHM phenotype to near-normal control levels. We conclude that abolishing conserved RNA splicing acceptor/donor sites and directing the splicing machinery to skip mutant or out-of-frame exons through myoediting allow correction of the cardiac abnormalities associated with DMD by eliminating the underlying genetic basis of the disease.


Prenatal diagnosis of a de novo ring chromosome 11.
Ring chromosomes are uncommon findings in prenatal diagnosis. Growth retardation is the most significant manifestation, in particular among patients with rings of larger chromosomes. A 30-year-old gravida 1, para 0 white woman was referred for genetic counseling because of maternal anxiety. Cytogenetic analysis of amniotic fluid cells at 16 weeks gestation revealed an abnormal mosaic female chromosome complement; 46,XX,r(11)(p15q25)[14]/45,XX,-11[7]. The ring 11 showed no detectable loss of chromosomal material at 450 band level. Both parents had a normal karyotype. Fluorescence in situ hybridization demonstrated intact subtelomeric regions in the ring chromosome. A targeted ultrasound evaluation at the time of consultation suggested no significant abnormalities. The parents were counseled and subsequently decided to terminate the pregnancy. The autopsy revealed an immature female fetus with abnormal craniofacial features including brachycephaly, low-set ears and hypertelorism, bicornuate uterus, and calcifications in the renal tubules. The abnormal phenotypes could be a consequence of the ring instability, submicroscopic deletion, and/or alteration of genetic material at the site of fusion.


Modifiable behaviors help to explain the inequalities in perceived health associated with deprivation and social class: evidence from a national sample.
The association between health and deprivation is of serious concern to many health promotion agencies. The purpose of the current study was to assess whether modifiable behaviors of physical activity (PA), sports participation, diet, smoking and body mass index (BMI) can help to explain these inequalities in a sample of 4653 respondents from Northern Ireland. The study is based on a cross-sectional survey of Northern Irish adults. Responses to a self-rated health question were dichotomized and binary logistic regression was used to identify the health inequalities between areas of high, middle or low deprivation. These differences were further adjusted for other sociodemographic factors and subsequently for various modifiable behaviors of PA, sports participation, diet, smoking, and BMI. Respondents from high and middle areas of deprivation are more likely to report poorer health. As soon as sociodemographic factors and other modifiable behaviors were included, these inequalities either disappeared or were greatly reduced. Many inequalities in health in NI can be explained by the respondents' sociodemographic characteristics that can be further explained by introducing information about respondents who meet the recommended PA guidelines, play sport, eat 5 portions of fruit and vegetables, and maintain an optimal BMI.


Accuracy of very low pretest probability estimates for pulmonary embolism using the method of attribute matching compared with the Wells score.
Attribute matching matches an explicit clinical profile of a patient to a reference database to estimate the numeric value for the pretest probability of an acute disease. The authors tested the accuracy of this method for forecasting a very low probability of venous thromboembolism (VTE) in symptomatic emergency department (ED) patients. The authors performed a secondary analysis of five data sets from 15 hospitals in three countries. All patients had data collected at the time of clinical evaluation for suspected pulmonary embolism (PE). The criterion standard to exclude VTE required no evidence of PE or deep venous thrombosis (DVT) within 45 days of enrollment. To estimate pretest probabilities, a computer program selected, from a large reference database of patients previously evaluated for PE, patients who matched 10 predictor variables recorded for each current test patient. The authors compared the outcome frequency of having VTE [VTE(+)] in patients with a pretest probability estimate of <2.5% by attribute matching, compared with a value of 0 from the Wells score. The five data sets included 10,734 patients, and 747 (7.0%, 95% confidence interval [CI] = 6.5% to 7.5%) were VTE(+) within 45 days. The pretest probability estimate for PE was <2.5% in 2,975 of 10,734 (27.7%) patients, and within this subset, the observed frequency of VTE(+) was 48 of 2,975 (1.6%, 95% CI = 1.2% to 2.1%). The lowest possible Wells score (0) was observed in 3,412 (31.7%) patients, and within this subset, the observed frequency of VTE(+) was 79 of 3,412 (2.3%, 95% CI = 1.8% to 2.9%) patients. Attribute matching categorizes over one-quarter of patients tested for PE as having a pretest probability of <2.5%, and the observed rate of VTE within 45 days in this subset was <2.5%.


SirT1 in muscle physiology and disease: lessons from mouse models.
Sirtuin 1 (SirT1) is the largest of the seven members of the sirtuin family of class III nicotinamide adenine dinucleotide (NAD(+))-dependent protein deacetylases, whose activation is beneficial for metabolic, neurodegenerative, inflammatory and neoplastic diseases, and augments life span in model organisms (Finkel et al., 2009; Lavu et al., 2008). In vitro studies show that SirT1 protects genome integrity and is involved in circadian physiological rhythms (Asher et al., 2008; Nakahata et al., 2008; Oberdoerffer et al., 2008). In the last few years, a fundamental role for SirT1 in the metabolism and differentiation of skeletal muscle cells has been uncovered (Fulco et al., 2003), and the use of specific transgenic or knockout SirT1 mouse models implicates it in the protection of heart muscle from oxidative and hypertrophic stresses (Alcendor et al., 2007). In this Perspective, we review the recent exciting findings that have established a key role for the 'longevity' protein SirT1 in skeletal and heart muscle physiology and disease. Furthermore, given the multiple biological functions of SirT1, we discuss the unique opportunities that SirT1 mouse models can offer to improve our integrated understanding of the metabolism, as well as the regeneration and aging-associated changes in the circadian function, of skeletal and heart muscle.


Targeting Plasmodium falciparum purine salvage enzymes: a look at structure-based drug development.
New antimalarials are needed due to the rapid development of resistance to currently deployed drugs. Because Plasmodium species are unable to synthesize purines, purine salvage pathways have been proposed as novel anti-malarial targets. The purine salvage pathway in Plasmodium is streamlined with adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and hypoxanthine-xanthine-guanine-phosphoribosyltransferase (HXGPRT) representing the major pathway for purine acquisition. Plasmodium falciparum enzymes PfADA and PfPNP have unique dual specificity that enable them to act upon methylthiopurines resulting from polyamine synthesis. Thus Plasmodium ADA and PNP function in both purine salvage and purine recycling. Genetic studies have confirmed the importance of Plasmodium PNP for viability of malaria parasites. Immucillins, powerful picomolar transition state inhibitors of PNP, are active against cultured Plasmodium falciparum and inhibit all Plasmodium PNPs tested. Several immucillins have undergone human clinical trials, and these compounds represent a new class of compounds with potential activity against human malarias.


Interplay of domain structure and phase transitions: theory, experiment and functionality.
Domain walls and phase boundaries are fundamental ingredients of ferroelectrics and strongly influence their functional properties. Although both interfaces have been studied for decades, often only a phenomenological macroscopic understanding has been established. The recent developments in experiments and theory allow to address the relevant time and length scales and revisit nucleation, phase propagation and the coupling of domains and phase transitions. This review attempts to specify regularities of domain formation and evolution at ferroelectric transitions and give an overview on unusual polar topological structures that appear as transient states and at the nanoscale. We survey the benefits, validity, and limitations of experimental tools as well as simulation methods to study phase and domain interfaces. We focus on the recent success of these tools in joint scale-bridging studies to solve long lasting puzzles in the field and give an outlook on recent trends in superlattices.


Mutation screening of CHD5 in melanoma-prone families linked to 1p36 revealed no deleterious coding or splice site changes.
A subset of cutaneous malignant melanoma and dysplastic nevi (CMM/DN) families is linked to 1p36. To date, no CMM/DN susceptibility gene has been identified at this locus. Data from mouse studies identified chromodomain helicase DNA binding protein 5 (CHD5) as a tumor suppressor affecting cellular proliferation and apoptosis via the CDKN2A/p53 pathway. Based on these findings, we felt it was important to screen CHD5 as a familial CMM/DN susceptibility gene. Eight unrelated CMM/DN families showing prior evidence of linkage to the 1p36 locus were identified for CHD5 mutation screening. One CMM/DN affected and one unaffected individual from each family were selected for sequencing of the CHD5 coding exons and their respective intron-exon boundaries. CHD5 variants that were identified solely among affecteds in the screening panel were further assessed by sequencing additional affected and unaffected members of these families to determine if the variant co-segregated with the CMM/DN phenotype. Single nucleotide polymorphisms in the CHD5 intronic and coding regions were identified among affecteds in the screening panel. None of these variants completely co-segregated with CMM/DN affection status among these eight families. There is no evidence to support CHD5 as a major melanoma susceptibility gene among the eight CMM/DN families screened.


Finger joint laxity, number of previous pregnancies and pregnancy induced back pain in a cohort study.
General joint hypermobility is estimated to affect about 10% of the population and is a prerequisite of heritable connective tissue disorders where fragile connective tissue is a prominent feature. Pregnancy induced back pain is common whereas about 10% of women still have disabling pain several years after childbirth. The pathogenesis of the pain condition is uncertain, although several risk factors are suggested including general joint hypermobility. In the present study, the possible association of peripheral joint mobility in early pregnancy on the incidence of back pain with onset during pregnancy and persisting after childbirth was explored. A cohort of 200 pregnant women recruited from antenatal health care clinics was assessed by questionnaire and clinical examination, including measurement of passive abduction of the left fourth finger, throughout pregnancy and at 13 weeks postpartum. Comparisons were made between women with and without back pain. Statistical tests used were χ2-test, t-test, Spearman correlation and multiple logistic regression. In the cohort, the mean passive abduction angle of the left fourth finger increased from 40.1° in early pregnancy to 41.8° at the postpartum appointment. At the postpartum appointment, women in the back pain group had a significantly larger mean passive abduction angle of the left fourth finger of 4.4°, twice as many previous pregnancies and deliveries, and more than twice as frequent back pain in previous pregnancy, as compared with women with no persistent back pain. A similar pattern was displayed in late pregnancy. In a multiple regression analysis, the passive abduction angle of the left fourth finger in early pregnancy and the number of previous pregnancies were positively, significantly and independently associated to the incidence of back pain in late pregnancy and postpartum. Finger joint laxity as a reflection of constitutional weakness of connective tissue and number of previous pregnancies were associated with the development of back pain induced in pregnancy and persisting after childbirth. These factors may provide a foundation for development of targeted prevention strategies, but this have to be confirmed in future research including measurement of general joint laxity.


mRNA Decapping and 5'-3' Decay Contribute to the Regulation of ABA Signaling in Arabidopsis thaliana.
Defects in RNA processing and degradation pathways often lead to developmental abnormalities, impaired hormonal signaling and altered resistance to abiotic and biotic stress. Here we report that components of the 5'-3' mRNA decay pathway, DCP5, LSM1-7 and XRN4, contribute to a proper response to a key plant hormone abscisc acid (ABA), albeit in a different manner. Plants lacking DCP5 are more sensitive to ABA during germination, whereas lsm1a lsm1b and xrn4-5 mutants are affected at the early stages of vegetative growth. In addition, we show that DCP5 and LSM1 regulate mRNA stability and act in translational repression of the main components of the early ABA signaling, PYR/PYL ABA receptors and SnRK2s protein kinases. mRNA decapping DCP and LSM1-7 complexes also appear to modulate ABA-dependent expression of stress related transcription factors from the AP2/ERF/DREB family that in turn affect the level of genes regulated by the PYL/PYR/RCAR-PP2C-SnRK2 pathway. These observations suggest that ABA signaling through PYL/PYR/RCAR receptors and SnRK2s kinases is regulated directly and indirectly by the cytoplasmic mRNA decay pathway.


Peroxisome proliferator-activated receptor alpha required for gene induction by dehydroepiandrosterone-3 beta-sulfate.
Peroxisome proliferator-activated receptor alpha (PPAR alpha) mediates the effects of foreign chemical peroxisome proliferators on liver and kidney, including the induction of peroxisomal, mitochondrial, and microsomal enzymes involved in beta-oxidation of fatty acids. However, the role of this receptor in the peroxisome proliferative effects of the endogenous steroid dehydroepiandrosterone (DHEA) is not known. DHEA-3 beta-sulfate fd(DHEA-S) is shown to induce a liver peroxisome proliferative response in rats in vivo at a dose at which DHEA is much less active, which is consistent with cultured hepatocyte studies indicating a requirement for sulfation for the activation of DHEA. Transient transfection experiments demonstrated that in contrast to the prototypic foreign chemical peroxisome proliferator pirinixic acid, DHEA-S and its 17 beta-reduced metabolite, 5-androstene-3 beta, 17 beta-diol-3 beta-sulfate, are inactive in mediating trans-activation by PPAR alpha in COS-1 cells. Two other mammalian PPAR isoforms, gamma and delta/Nucl, were also inactive with respect to DHEA-S trans-activation. To test whether PPAR alpha mediates peroxisomal gene induction by DHEA-S in intact animals, we administered DHEA-S or clofibrate to mice lacking a functional PPAR alpha gene. Both peroxisome proliferators markedly increased hepatic expression of two microsomal cytochrome P450 4A proteins as well as six mRNAs known to be associated with the peroxisomal proliferative response in wild-type mice. In contrast, neither DHEA-S nor clofibrate induced these hepatic proteins and mRNAs in PPAR alpha-deficient mice. Clofibrate-induced expression of kidney CYP4A mRNAs was also blocked in the PPAR alpha gene knockout mice. Thus, despite its unresponsiveness to steroidal peroxisome proliferators in transfection assays, PPAR alpha is obligatory for DHEA-S-stimulated hepatic peroxisomal gene induction. DHEA-S, or one of its metabolites, may thus serve as an important endogenous regulator of liver peroxisomal enzyme expression via a PPAR alpha-mediated pathway.


Prison Meditation Movements and Mass Incarceration.
By some estimates, more than half of inmates held in jails and prisons in the United States have a substance use disorder. Treatments involving the teaching of meditation and other contemplative practices have been developed for a variety of physical and mental disorders, including drug and alcohol addiction. At the same time, an expanding volunteer movement across the country has been bringing meditation and yoga into jails and prisons. This review first examines the experimental research on one such approach-mindfulness meditation as a treatment for drug and alcohol addiction, as well as the research on mindfulness in incarcerated settings. We argue that to make a substantial impact on recidivism, such programs must mirror volunteer programs which emphasize interdependency and non-duality between the "helper" and the "helped," and the building of meditation communities both inside and outside of prison.


Effects of supplementing n-3 fatty acid enriched eggs and walnuts on cardiovascular disease risk markers in healthy free-living lacto-ovo-vegetarians: a randomized, crossover, free-living intervention study.
Plant and marine n-3 fatty acids (FA) may favorably modify select markers of cardiovascular disease risk. Whether supplementing the habitual diet of lacto-ovo-vegetarians (LOV) with walnuts (containing α-linolenic acid, ALA) and n-3 FA enriched eggs (containing primarily docosahexaenoic acid, DHA and ALA) would have equivalent effects on CVD risk factors is explored in this study. In this study, 20 healthy free-living LOVs following their habitual diet were randomly assigned in a crossover design to receive one of three supplements: n-3 FA enriched egg (6/week), walnuts (28.4 g, 6/week) or a standard egg, 6/week (control) for 8 weeks each with 4-wk washout between treatments. Erythrocyte membrane fatty acids, serum lipids and inflammatory markers were measured at the end of each treatment. Dietary compliance was observed by an expected increase in erythrocyte membrane ALA following the walnut treatment and in DHA following the n-3 FA enriched egg treatment. Walnut treatment lowered serum triacylglycerol, total cholesterol and Apo B (p < 0.05) compared to the standard egg but not the n-3 FA enriched egg treatment. However, walnut treatment significantly reduced total: HDL cholesterol ratio compared to both egg treatments. There were no differences between treatments for any of the inflammatory markers. For LOV, a direct source of DHA such as n-3 FA enriched eggs seems necessary to increase membrane levels of DHA. However for producing an overall favorable blood lipid profile, daily consumption of a handful of walnuts rich in ALA may be a preferred option for lacto-ovo vegetarian.


The duration of intrathecal bupivacaine mixed with lidocaine.
Although spinal bupivacaine may have too long duration to be useful in the ambulatory setting, recent animal data suggest that lidocaine added to spinal bupivacaine may reduce the duration of bupivacaine spinal anesthesia. We explored whether lidocaine added to spinal bupivacaine could shorten the duration of bupivacaine spinal anesthesia in humans similarly to what has been reported in animals. Ninety patients presenting for transurethral resection of bladder tumor or prostate were assigned to one of three groups by double blind randomization to receive intrathecal 1.5 mL of hyperbaric 0.5% bupivacaine, plus 0.6 mL of one of three solutions: saline (Group I, n = 30, control), 1% lidocaine (Group II, n = 30), and 2% lidocaine (Group III, n = 30). Peak sensory block level, time to peak sensory block, times to two-segment, L1, and S2 regressions from peak sensory block, motor blocks at peak sensory block, L1, and S2 regressions, and postanesthesia care unit stay time (PACU time) were measured. Times to peak sensory block were similar in all three groups. Times to two-segment, L1, and S2 regressions from peak sensory block, and PACU time were significantly reduced in Group II compared to Group I. Times to L1, S2 regressions, and PACU times in Group III were significantly prolonged. We conclude that lidocaine (6 mg) mixed to spinal bupivacaine (7.5 mg) can shorten the duration of bupivacaine spinal anesthesia, therefore provide more rapid recovery from the spinal anesthesia compared to the same dose of bupivacaine (7.5 mg) alone.


Exposure to radiofrequency radiation increases the risk of breast cancer: A systematic review and meta-analysis.
The present systematic review and meta-analysis investigated the association between exposure to radiofrequency radiation and the risk of breast cancer. The published studies that were available in PubMed, Embase, Cochrane Library, Ovid MEDLINE, CINAHL Plus, Web of Science, Airiti Library, Networked Digital Library of Theses and Dissertations and ProQuest until May 2020 were investigated. A total of eight studies (four case-control and four cohort studies) were eligible for quantitative analysis. A significant association between radiofrequency radiation exposure and breast cancer risk was detected [pooled relative risk (RR)=1.189; 95% confidence interval (CI), 1.056-1.339]. Subgroup analyses indicated that radiofrequency radiation exposure significantly increased the risk of breast cancer susceptibility among subjects aged ≥50 years (RR=2.179; 95% CI, 1.260-3.770). Pooled estimates revealed that the use of electrical appliances, which emit radiofrequency radiation, such as mobile phones and computers, significantly increased breast cancer development (RR=2.057; 95% CI, 1.272-3.327), while occupational radiofrequency exposure and transmitters did not increase breast cancer development (RR=1.274; 95% CI, 0.956-1.697; RR=1.133; 95% CI, 0.987-1.300, respectively). It was concluded that radiofrequency radiation exposure significantly increased the risk of breast cancer, especially in women aged ≥50 years and in individuals who used electric appliances, such as mobile phones and computers. In accordance with Preferred Reporting Items for Systematic Reviews and Meta-analysis, an evaluation protocol was prepared and registered with the PROSPERO database (registration no. CRD42018087283).


Persistence of the uncanny valley: the influence of repeated interactions and a robot's attitude on its perception.
The uncanny valley theory proposed by Mori has been heavily investigated in the recent years by researchers from various fields. However, the videos and images used in these studies did not permit any human interaction with the uncanny objects. Therefore, in the field of human-robot interaction it is still unclear what, if any, impact an uncanny-looking robot will have in the context of an interaction. In this paper we describe an exploratory empirical study using a live interaction paradigm that involved repeated interactions with robots that differed in embodiment and their attitude toward a human. We found that both investigated components of the uncanniness (likeability and eeriness) can be affected by an interaction with a robot. Likeability of a robot was mainly affected by its attitude and this effect was especially prominent for a machine-like robot. On the other hand, merely repeating interactions was sufficient to reduce eeriness irrespective of a robot's embodiment. As a result we urge other researchers to investigate Mori's theory in studies that involve actual human-robot interaction in order to fully understand the changing nature of this phenomenon.


Therapeutic relationships in vocational rehabilitation: predicting good relationships for people with psychosis.
Therapeutic relationships between clients and vocational rehabilitation workers have been shown to predict entering competitive employment. We aimed to determine predictors of good relationships, using data from an international randomized controlled trial of supported employment (n=312). Baseline predictors of early therapeutic relationships with vocational workers were assessed, along with the impact of vocational status and changing clinical and social functioning variables on relationship ratings over time. Associations between client and professional relationship ratings were also explored. Better early client-rated therapeutic relationship was predicted by better baseline relationship with the clinical keyworker, being in the Individual Placement and Support (IPS) service, the absence of work history and a greater proportion of care needs being met, whereas over time it was predicted by being in the IPS service. Professional-rated early relationship was predicted by social disability and remission, while over time it was predicted by being the same sex as the client, duration of the relationship and the client's increasing anxiety. Client and professional ratings were positively associated but clients' ratings were higher than professionals', particularly in the IPS service. Relationships were better where clients may have been more motivated to engage, including by their prior experience of a good therapeutic relationship with the clinical keyworker.


UV-induced drug release from photoactive REV sensitized by suprofen.
In order to achieve local administration of drugs, calcein (CAL) encapsulated reverse phase evaporation vesicles (REV) carrying photoactive destabilization agent suprofen (SPF) in the lipid bilayer were prepared. Effect of both UV-A and UV-B photoactivation of liposomal membrane incorporated SPF on the destabilization of the liposome bilayer and the release of encapsulated CAL was investigated. Standard REV of phosphatidylcholine (PC):cholesterol (CHOL) in 7:3 molar ratio, and photoactive REV of PC:CHOL:SPF, and DPPC:CHOL:SPF in 7:3:3 molar ratio were prepared. CAL encapsulation efficiency (EE (%)) and in situ release was studied. SPF incorporation in the PC REV membrane led to approximately 5% increase in the EE (34%) in comparison to standard REV (29%). EE decreased (21%) when DPPC was used to replace PC. Exposure to UV-B caused the highest CAL release. The lowest release was from the unexposed REV. DPPC led to a higher liposomal membrane stability (lower CAL release) than PC. A linear relationship was observed between UV-B exposure duration and REV permeability. This study revealed that membrane destabilization of SPF incorporated REV was best achieved upon photoactivation of the membrane-localized SPF by a 40 min exposure to UV-B.


Contextual factors related to the drinking behaviors of American business and professional women.
American women in business and professional occupations (n = 453) completed a survey that included questions on alcohol use, drinking context, and work and other activities. Spouses' and best friends' consumption and subjects' frequency in drinking settings correlated with their consumption and negative consequences of alcohol use. Multiple regression analyses indicated that predictor variables for consumption were frequency in drinking settings, a measure of three personal motives for drinking, age, and number of organization memberships. Predictor variables for negative consequences were subject's consumption, spouse's drinking, frequency in drinking settings, and age. The results suggest that social context may be important in understanding women's drinking. Variables directly related to drinking, such as time spent in drinking situations, are correlated with increased drinking, while other contextual variables, such as membership in organizations, may play a preventive role.


Rowers' high: behavioural synchrony is correlated with elevated pain thresholds.
Physical exercise is known to stimulate the release of endorphins, creating a mild sense of euphoria that has rewarding properties. Using pain tolerance (a conventional non-invasive assay for endorphin release), we show that synchronized training in a college rowing crew creates a heightened endorphin surge compared with a similar training regime carried out alone. This heightened effect from synchronized activity may explain the sense of euphoria experienced during other social activities (such as laughter, music-making and dancing) that are involved in social bonding in humans and possibly other vertebrates.


Increased Alcohol Seeking in Mice Lacking Gpr88 Involves Dysfunctional Mesocorticolimbic Networks.
Alcohol use disorder (AUD) is devastating and poorly treated, and innovative targets are actively sought for prevention and treatment. The orphan G protein-coupled receptor GPR88 is enriched in mesocorticolimbic pathways, and Gpr88 knockout mice show hyperactivity and risk-taking behavior, but a potential role for this receptor in drug abuse has not been examined. We tested Gpr88 knockout mice for alcohol-drinking and -seeking behaviors. To gain system-level understanding of their alcohol endophenotype, we also analyzed whole-brain functional connectivity in naïve mice using resting-state functional magnetic resonance imaging. Gpr88 knockout mice showed increased voluntary alcohol drinking at both moderate and excessive levels, with intact alcohol sedation and metabolism. Mutant mice also showed increased operant responding and motivation for alcohol, while food and chocolate operant self-administration were unchanged. Alcohol place conditioning and alcohol-induced dopamine release in the nucleus accumbens were decreased, suggesting reduced alcohol reward in mutant mice that may partly explain enhanced alcohol drinking. Seed-based voxelwise functional connectivity analysis revealed significant remodeling of mesocorticolimbic centers, whose hallmark was predominant weakening of prefrontal cortex, ventral tegmental area, and amygdala connectional patterns. Also, effective connectivity from the ventral tegmental area to the nucleus accumbens and amygdala was reduced. Gpr88 deletion disrupts executive, reward, and emotional networks in a configuration that reduces alcohol reward and promotes alcohol seeking and drinking. The functional connectivity signature is reminiscent of alterations observed in individuals at risk for AUD. The Gpr88 gene, therefore, may represent a vulnerability/resilience factor for AUD, and a potential drug target for AUD treatment.


DNA-directed artificial light-harvesting antenna.
Designing and constructing multichromophoric, artificial light-harvesting antennas with controlled interchromophore distances, orientations, and defined donor-acceptor ratios to facilitate efficient unidirectional energy transfer is extremely challenging. Here, we demonstrate the assembly of a series of structurally well-defined artificial light-harvesting triads based on the principles of structural DNA nanotechnology. DNA nanotechnology offers addressable scaffolds for the organization of various functional molecules with nanometer scale spatial resolution. The triads are organized by a self-assembled seven-helix DNA bundle (7HB) into cyclic arrays of three distinct chromophores, reminiscent of natural photosynthetic systems. The scaffold accommodates a primary donor array (Py), secondary donor array (Cy3) and an acceptor (AF) with defined interchromophore distances. Steady-state fluorescence analyses of the triads revealed an efficient, stepwise funneling of the excitation energy from the primary donor array to the acceptor core through the intermediate donor. The efficiency of excitation energy transfer and the light-harvesting ability (antenna effect) of the triads was greatly affected by the relative ratio of the primary to the intermediate donors, as well as on the interchromophore distance. Time-resolved fluorescence analyses by time-correlated single-photon counting (TCSPC) and streak camera techniques further confirmed the cascading energy transfer processes on the picosecond time scale. Our results clearly show that DNA nanoscaffolds are promising templates for the design of artificial photonic antennas with structural characteristics that are ideal for the efficient harvesting and transport of energy.


Peripheral atherosclerosis and serum lipoprotein(a) in diabetes.
Serum lipoprotein(a) [Lp(a)] is strongly associated with atherosclerosis in nondiabetic individuals. To see if atherosclerosis is also associated with serum Lp(a) in both IDDM and NIDDM, we determined the correlation between the toe systolic blood pressure index (TSPI) and serum Lp(a) in tightly controlled diabetic patients without nephropathy. Cross-sectional study of 57 IDDM and 35 NIDDM patients. All patients had been under strict glycemic control for at least 6 months. The main outcome measure was TSPI of both lower extremities. In addition, we measured serum Lp(a) and other serum lipids, serum uric acid, total plasma homocysteine, plasma C-peptide, HbA1c, albumin excretion rate, glomerular filtration rate, BMI, abdominal fat distribution, left ventricular hypertrophy, probabilities for cardiovascular disease (CVD), and routine clinical parameters. TSPI was closely and independently related to serum Lp(a) in IDDM patients: R2 = 0.2999, partial P = 0.0005, and in NIDDM patients: R2 = 0.7326, partial P = 0.0030. TSPI was associated with symptoms of CVD. Median serum Lp(a) concentration was normal in IDDM (45 mg/l [range 10-870]) and NIDDM (72 mg/l [11-803]) patients. Systemic atherosclerosis measured as the degree of peripheral occlusive arterial disease is strongly associated with serum Lp(a) in both IDDM and NIDDM patients. Serum Lp(a), however, is normal in both types of diabetic patients. Thus, it is indicated that serum Lp(a) should be measured in diabetic patients when assessing their risk profile for atherosclerosis.


Genetic evidence that the meiotic recombination hotspot at the HIS4 locus of Saccharomyces cerevisiae does not represent a site for a symmetrically processed double-strand break.
In the yeast Saccharomyces cerevisiae, the binding of the Rap1 protein to a site located between the 5' end of the HIS4 gene and the 3' end of BIK1 stimulates meiotic recombination at both flanking loci. By using strains that contain mutations located in HIS4 and BIK1, we found that most recombination events stimulated by the binding of Rap1 involve HIS4 or BIK1, rather than bidirectional events including both loci. The patterns of aberrant segregation indicate that most of the Rap1-stimulated recombination events do not represent the symmetric processing of a double-strand DNA break located at the Rap1-binding site.


Identity Conflict and Sexual Risk for Black and Latino YMSM.
Young (aged 18-30) Black and Latino men who have sex with men are at a higher risk of contracting HIV than their White counterparts. In order to better understand the unique nature of sexual risk-taking, we examined the extent to which ethnic group, ethnic identity, and sexual pride predicted condomless anal sex with casual partners among 161 young men who have sex with men (YMSM) who identify as Black or Latino. Negative binomial regressions were conducted using a cross-sectional design. Sexual pride was a negative predictor of condomless anal sex across all participants, but this effect was moderated by ethnic exploration and ethnic group; the relationship between sexual pride and condomless anal sex was strengthened by greater ethnic exploration among Latino YMSM, and weakened by greater ethnic exploration among Black YMSM. Implications for intersectional identity, identity conflict, and HIV prevention among young gay men of color are discussed.


From drug delivery systems to drug release, dissolution, IVIVC, BCS, BDDCS, bioequivalence and biowaivers.
This is a summary report of the conference on drug absorption and bioequivalence issues held in Titania Hotel in Athens (Greece) from the 28(th) to the 30(th) of May 2009. The conference included presentations which were mainly divided into three sections. The first section focused on modern drug delivery systems such as polymer nanotechnology, cell immobilization techniques to deliver drugs into the brain, nanosized liposomes used in drug eluting stents, encapsulation of drug implants in biocompatible polymers, and application of differential scanning calorimetry as a tool to study liposomal stability. The importance of drug release and dissolution were also discussed by placing special emphasis on camptothecins and oral prolonged release formulations. The complexity of the luminal environment and the value of dissolution in lyophilized products were also highlighted. The second session of the conference included presentations on the Biopharmaceutics Classification Scheme (BCS), the Biopharmaceutics Drug Disposition Classification System (BDDCS), and the role of transporters in the classification of drugs. The current status of biowaivers and a modern view on non-linear in vitro-in vivo (IVIVC) correlations were also addressed. Finally, this section ended with a special topic on biorelevant dissolution media and methods. The third day of the conference was dedicated to bioequivalence. Emphasis was placed on high within-subject variability and its impact on study design. Two unresolved issues of bioequivalence were also discussed: the use of generic antiepileptic drugs and the role of metabolites in bioequivalence assessment. Finally, the conference closed with a presentation of the current regulatory status of WHO and EMEA.


F₁ cross-breed between susceptible BALB/c and resistant Swiss mice infected with Leishmania major exhibit an intermediate phenotype for lesion sizes and type 1 cytokines but show low level of total IgG antibodies.
Our current understanding of the host immune response during leishmaniases largely derives from studies performed in mice due to the intrusive techniques required to study infected human patients. Swiss mice are highly resistant to Leishmania infections in concordance with observed response in humans, while BALB/c mice indicate a high-susceptibility phenotype. Developing a cross-breed between BALB/c and Swiss mice may have important consequences on disease development, immune responses and parasite killing, as yet, response of the cross-breed to Leishmania infection is superficial. The aim of the present study was to determine disease course and immune responses in F₁ cross-breed between BALB/c and Swiss albino mice infected with L. major. Three mice groups were infected intradermally with stationary-phase L. major parasites with parental strains (BALB/c and Swiss albino) as controls. Lesion development was monitored weekly for 8 weeks and monocyte chemotactic protein (MCP-1), macrophage inflammatory protein (MIP-1α), interferon-gamma (IFN-γ) and IgG antibody quantified by enzyme-linked immunosorbent assay. The data were analysed using one-way analysis of variance and Tukey-Kramer test. Results indicated F₁ mice having intermediate lesion sizes, type 1 cytokine levels and footpad parasite loads as compared to the parental strains. However, the F₁ mice had low levels of IgG antibodies and parasite burden in the spleen. (P < 0.05). This study concludes that the F₁ cross-breed between resistant and susceptible mice may be used as a requisite model to study the role of genetics in leishmaniases and perhaps other intracellular parasites.


Stratification substantially reduces behavioral variability in the hypoxic-ischemic stroke model.
Stroke is the most common cause of long-term disability, and there are no known drug therapies to improve recovery after stroke. To understand how successful recovery occurs, dissect candidate molecular pathways, and test new therapies, there is a need for multiple distinct mouse stroke models, in which the parameters of recovery after stroke are well defined. Hypoxic-ischemic stroke is a well-established stroke model, but behavioral recovery in this model is not well described. We therefore examined a panel of behavioral tests to see whether they could be used to quantify functional recovery after hypoxic-ischemic stroke. We found that in C57BL/6J mice this stroke model produces high mortality (approximately one-third) and variable stroke sizes, but is fast and easy to perform on a large number of mice. Horizontal ladder test performance on day 1 after stroke was highly and reproducibly correlated with stroke size (P < 0.0001, R(2) = 0.7652), and allowed for functional stratification of mice into a group with >18% foot faults and 2.1-fold larger strokes. This group exhibited significant functional deficits for as long as 3 weeks on the horizontal ladder test and through the last day of testing on automated gait analysis (33 days), rotarod (30 days), and elevated body swing test (EBST) (36 days). No deficits were observed in an automated activity chamber. We conclude that stratification by horizontal ladder test performance on day 1 identifies a subset of mice in which functional recovery from hypoxic-ischemic stroke can be studied.


Spiders spinning electrically charged nano-fibres.
Most spider threads are on the micrometre and sub-micrometre scale. Yet, there are some spiders that spin true nano-scale fibres such as the cribellate orb spider, Uloborus plumipes. Here, we analyse the highly specialized capture silk-spinning system of this spider and compare it with the silk extrusion systems of the more standard spider dragline threads. The cribellar silk extrusion system consists of tiny, morphologically basic glands each terminating through exceptionally long and narrow ducts in uniquely shaped silk outlets. Depending on spider size, hundreds to thousands of these outlet spigots cover the cribellum, a phylogenetically ancient spinning plate. We present details on the unique functional design of the cribellate gland-duct-spigot system and discuss design requirements for its specialist fibrils. The spinning of fibres on the nano-scale seems to have been facilitated by the evolution of a highly specialist way of direct spinning, which differs from the aqua-melt silk extrusion set-up more typical for other spiders.


Physiological Capacity and Training Routines of Elite Cross-Country Skiers: Approaching the Upper Limits of Human Endurance.
Cross-country (XC) skiing is one of the most demanding of endurance sports, involving protracted competitions on varying terrain employing a variety of skiing techniques that require upper- and/or lower-body work to different extents. Through more effective training and extensive improvements in equipment and track preparation, the speed of cross-country ski races has increased more than that of any other winter Olympic sport, and, in addition, new types of racing events have been introduced. To a certain extent this has altered the optimal physiological capacity required to win, and the training routines of successful skiers have evolved accordingly. The long-standing tradition of researchers working closely with XC-ski coaches and athletes to monitor progress, improve training, and refine skiing techniques has provided unique physiological insights revealing how these athletes are approaching the upper limits of human endurance. This review summarizes current scientific knowledge concerning the demands involved in elite XC skiing, as well as the physiological capacity and training routines of the best athletes.


The expanded bed biofilter: combined nitrification, solids destruction, and removal of bacteria.
Developed for tertiary nitrification, this biofilter also removed carbonaceous BOD (cBOD) and (SS). Because the biofilter is expanded, it cannot clog, and therefore does not require backflushing; yet, it removed a significant proportion of the influent SS. This unanticipated capability was due to the activities of heterotrophic bacteria, protozoa, and metazoa (nematode and oligochaete worms). The expanded bed is an intensified process, which is based on natural immobilization of microbes to small support particles. Using glassy coke as the support material, an attached layer of microbes develops, forming particulate biofilms having a superficial surface area of 1 800 m2 m(-3)(expandedbed). Autotrophic nitritifiers (Nitrosomonas spp.) were detected in the biofilm using rRNA-based molecular methods and were likely responsible, at least in part, for reducing the ammonia concentration by up to 99% (to 0.1 mg L(-1)), while the other organisms reduced cBOD and SS by up to 56% and 62%, respectively. Furthermore, the influent concentrations of Escherichia coli, coliform and heterotrophic bacteria were reduced by over 80%. It thereby provides a single process solution for combined tertiary nitrification and solids removal. Operating the process to consistently achieve < 0.5mg NH3N L(-1) and at the same time removing a significant fraction of cBOD and SS, it can replace processes such as SAFs or NTFs followed by a sandfilter.


Effect of mask-roughness on printed contact-size variation in extreme-ultraviolet lithography.
Relying on reflective mask technology, extreme-ultraviolet (EUV) lithography is particularly vulnerable to mask substrate roughness. Previous research has shown mask roughness to play a significant role in printed line-edge roughness (LER). Here the analysis of mask-roughness effects is extended to printed contact-size variations. Unlike LER, illumination partial coherence is found to have little affect on the results for contacts that are near the diffraction limit. Analysis shows that, given the current state-of-the-art EUV mask, mask roughness has a significant effect on the process window for small contacts. The analysis also shows that a significant portion of the contact-size variation observed in recent 0.1-numerical-aperture EUV exposures can be attributed to the mask-roughness effect studied here.


Identification and functional characterization of cytoplasmic determinants of plasmid DNA nuclear import.
Import of exogenous plasmid DNA (pDNA) into mammalian cell nuclei represents a key intracellular obstacle to efficient non-viral gene delivery. This includes access of the pDNA to the nuclei of non-dividing cells where the presence of an intact nuclear membrane is limiting for gene transfer. Here we identify, isolate, and characterize, cytoplasmic determinants of pDNA nuclear import into digitonin-permeabilized HeLa cells. Depletion of putative DNA-binding proteins, on the basis of their ability to bind immobilized pDNA, abolished pDNA nuclear import supporting the critical role of cytoplasmic factors in this process. Elution of pDNA-bound proteins, followed by two-dimensional sodium dodecyl polyacrylamide gel electrophoresis identified several candidate DNA shuttle proteins. We show that two of these, NM23-H2, a ubiquitous c-Myc transcription-activating nucleoside diphosphate kinase, and the core histone H2B can both reconstitute pDNA nuclear import. Further, we demonstrate a significant increase in gene transfer in non-dividing HeLa cells transiently transfected with pDNA containing binding sequences from two of the DNA shuttle proteins, NM23-H2 and the homeobox transcription factor Chx10. These data support the hypothesis that exogenous pDNA binds to cytoplasmic shuttle proteins and is then translocated to the nucleus using the minimal import machinery. Importantly, increasing the binding of pDNA to shuttle proteins by re-engineering reporter plasmids with shuttle binding sequences enhances gene transfer. Increasing the potential for exogenously added pDNA to bind intracellular transport cofactors may enhance the potency of non-viral gene transfer.


Changes over 14 years in androgenicity and body mass index in a biracial cohort of reproductive-age women.
Body mass index (BMI) is directly related to testosterone (total T and free T) and inversely to SHBG cross-sectionally, but little is known about how changes in body fat and androgen markers affect each other over time. Participants included 969 White and Black women from the Coronary Artery Risk Development in Young Adults (CARDIA) cohort, who were ages 18-30 at entry into the study and were pre- or perimenopausal 16 yr later at the time of the CARDIA Women's Study (CWS). Total T and SHBG were assayed from specimens drawn at the CWS examination and stored serum from the yr 2 and 10 CARDIA exams. Free T was calculated based on total T and SHBG. BMI and waist circumference were measured at yr 2, 10, and 16. Despite clinically significant increases in BMI and waist circumference, total T and free T tended to decline, whereas SHBG remained relatively constant. BMI and waist circumference were directly correlated with free T and inversely correlated with SHBG in cross-sectional analyses. In longitudinal, multivariable analyses, an annualized increase in BMI was inversely related to a concurrent annualized decrease in SHBG (beta = -0.79 ng/dl, and se = 0.22 in Blacks; beta = -1.07 ng/dl; and se = 0.31 in Whites). However, early increases in BMI were not related to later decreases in SHBG. Increases in adiposity are closely tied to decreases in SHBG, but changes in BMI and SHBG may occur concurrently rather than sequentially.


Aging and luminance-adaptation effects on spatial contrast sensitivity.
Contrast sensitivity as a function of target luminance for four spatial frequencies (0.5, 2, 4, and 8 cycles/deg) was measured in younger (n = 12; age range, 19-35 years) and older (n = 11; age range, 68-79 years) adults in order to examine the feasibility of optical and neural explanations for the impairment of contrast sensitivity in older adults. All subjects were free from identifiable ocular disease and had good acuity. Sensitivity for each spatial frequency was measured at eight luminance levels spanning 3.5 log units in the photopic-mesopic range. When gratings were flickered at 0.5 Hz, functions for older adults were displaced downward on the sensitivity axis across all luminance levels, and the slopes of these functions were steeper than those for younger adults, suggesting that optical mechanisms alone cannot account for the vision loss in older adults. Further measurements, in which spatial targets were flickered at 7.5 Hz, indicated that this faster temporal modulation affected sensitivity as a function of luminance differentially in younger and older adults. These data imply that the neural mechanisms subserving human spatial vision undergo significant changes during adulthood.


Stratification of Colloidal Particles on a Surface: Study by a Colloidal Probe Atomic Force Microscopy Combined with a Transform Theory.
Colloidal probe atomic force microscopy (CP-AFM) can be used for measuring force curves between the colloidal probe and the substrate in a colloidal suspension. In the experiment, an oscillatory force curve reflecting the layer structure of the colloidal particles on the substrate is usually obtained. However, the force curve is not equivalent to the interfacial structure of the colloidal particles. In this paper, the force curve is transformed into the number density distribution of the colloidal particles as a function of the distance from the substrate surface using our newly developed transform theory. It is found by the transform theory that the interfacial stratification is enhanced by an increase in an absolute value of the surface potential of the colloidal particle, despite a simultaneous increase in a repulsive electrostatic interaction between the substrate and the colloidal particle. To elucidate the mechanism of the stratification, an integral equation theory is employed. It is found that crowding of the colloidal particles in the bulk due to the increase in the absolute value of the surface potential of the colloidal particle leads to pushing out some colloidal particles to the wall. The combined method of CP-AFM and the transform theory (the experimental-theoretical study of the interfacial stratification) is related to colloidal crystallization, glass transition, and aggregation on a surface. Thus, the combined method is important for developments of colloidal nanotechnologies.


The controlling role of ATM in homologous recombinational repair of DNA damage.
The human genetic disorder ataxia telangiectasia (A-T), caused by mutation in the ATM gene, is characterized by chromosomal instability, radiosensitivity and defective cell cycle checkpoint activation. DNA double-strand breaks (dsbs) persist in A-T cells after irradiation, but the underlying defect is unclear. To investigate ATM's interactions with dsb repair pathways, we disrupted ATM along with other genes involved in the principal, complementary dsb repair pathways of homologous recombination (HR) or non-homologous end-joining (NHEJ) in chicken DT40 cells. ATM(-/-) cells show altered kinetics of radiation-induced Rad51 and Rad54 focus formation. Ku70-deficient (NHEJ(-)) ATM(-/-) chicken DT40 cells show radiosensitivity and high radiation-induced chromosomal aberration frequencies, while Rad54-defective (HR(-)) ATM(-/-) cells show only slightly elevated aberration levels after irradiation, placing ATM and HR on the same pathway. These results reveal that ATM defects impair HR-mediated dsb repair and may link cell cycle checkpoints to HR activation.


ISA-2011B, a Phosphatidylinositol 4-Phosphate 5-Kinase α Inhibitor, Impairs CD28-Dependent Costimulatory and Pro-inflammatory Signals in Human T Lymphocytes.
Phosphatidylinositol 4,5-biphosphate (PIP2) is a membrane phospholipid that controls the activity of several proteins regulating cytoskeleton reorganization, cytokine gene expression, T cell survival, proliferation, and differentiation. Phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) are the main enzymes involved in PIP2 biosynthesis by phosphorylating phosphatidylinositol 4-monophosphate (PI4P) at the D5 position of the inositol ring. In human T lymphocytes, we recently found that CD28 costimulatory molecule is pivotal for PIP2 turnover by recruiting and activating PIP5Kα. We also found that PIP5Kα is the main regulator of both CD28 costimulatory signals integrating those delivered by TCR as well as CD28 autonomous signals regulating the expression of pro-inflammatory genes. Given emerging studies linking alterations of PIP2 metabolism to immune-based diseases, PIP5Kα may represent a promising target to modulate immunity and inflammation. Herewith, we characterized a recently discovered inhibitor of PIP5Kα, ISA-2011B, for its inhibitory effects on T lymphocyte functions. We found that the inhibition of PIP5Kα lipid-kinase activity by ISA-2011B significantly impaired CD28 costimulatory signals necessary for TCR-mediated Ca2+ influx, NF-AT transcriptional activity, and IL-2 gene expression as well as CD28 autonomous signals regulating the activation of NF-κB and the transcription of pro-inflammatory cytokine and chemokine genes. Moreover, our data on the inhibitory effects of ISA-2011B on CD28-mediated upregulation of inflammatory cytokines related to Th17 cell phenotype in type 1 diabetes patients suggest ISA-2011B as a promising anti-inflammatory drug.


Effect of human placental extract on menopausal symptoms, fatigue, and risk factors for cardiovascular disease in middle-aged Korean women.
In Korea, human placental extract (HPE) has recently been used to treat various diseases (chronic liver diseases, menopause syndrome, chronic fatigue, skin pigment diseases, etc.), but evidence-based studies are not yet sufficient. The aim of this study was to examine the effects of HPE on menopausal symptoms, fatigue, and risk factors for cardiovascular disease in middle-aged Korean women in a randomized controlled trial. Korean women, aged 40 to 64 years, with menopausal symptoms and fatigue were recruited as participants. The women were randomly assigned to a placebo group or an HPE group. The HPE group received subcutaneous injections of HPE in the abdomen for 8 weeks, whereas the placebo group received normal saline. Then, the Menopause Rating Scale, and Fatigue Severity Scale, and Visual Analog Scale were administered, and risk factors for cardiovascular disease were assessed. The Menopause Rating Scale total baseline score was not different between the two groups; however, the score of the HPE group decreased significantly at 8 weeks compared with that of the placebo group (P = 0.033). Fatigue Severity Scale and Visual Analog Scale scores of the placebo group did not change, whereas the scores of the HPE group decreased significantly during the study period (Fatigue Severity Scale, P = 0.002; Visual Analog Scale, P = 0.001). The baseline 17beta-estradiol level was not significantly different between the two groups, but the 17beta-estradiol level of the HPE group was significantly increased at 8 weeks compared with that of the placebo group (P = 0.031). No changes in risk factors for cardiovascular disease were observed in either group. Menopausal symptoms and fatigue in middle-aged Korean women improved after 8 weeks of HPE treatment, whereas risk factors for cardiovascular disease did not change during the study period.


Acceptor sites for the oestrogen receptor in hen oviduct chromatin.
Partially purified hen oviduct oestrogen receptors, charged with [3H]oestradiol, were shown to specifically bind in vitro to purified hen oviduct chromatin. Maximal binding occurred within 60min at 0 degrees C in a Tris buffer containing 0.1 M-KCl and 0.5 mM-phenylmethanesulphonyl fluoride. The binding of the [3H]oestradiol-receptor complexes to intact purified chromatin was saturable, whereas the receptor binding to hen DNA remained linear. Saturation was further demonstrated by the minimal acceptor binding of receptor charged with [3H]oestradiol plus 200-fold oestradiol compared with [3H]oestradiol receptors at equal [3H]oestradiol concentrations. Scatchard analysis of [3H]oestradiol-receptor binding to chromatin above DNA levels gave indications of high-affinity binding with a low capacity. Further, the nuclear binding was tissue-specific since the binding to hen spleen chromatin was negligible. To further uncover the specific acceptor sites, proteins were removed from hen oviduct chromatin by increasing concentrations of guanidine hydrochloride (1-7M). Those residual fractions extracted with 3-7 M-guanidine hydrochloride had the highest acceptor activity (above DNA levels) with the peak activity uncovered by 5 M-guanidine hydrochloride. To further characterize the oestrogen-receptor acceptor sites, oviduct chromatin was bound to hydroxyapatite in the presence of 3 M-NaCl and then protein fractions were extracted sequentially with 1-7 M-guanidine hydrochloride. Each fraction was then reconstituted to pure hen DNA by reverse gradient dialysis. [3H]Oestradiol receptors were found to bind to the greatest degree to the fraction reconstituted from the 5 M-guanidine hydrochloride protein extract. Reconstituted nucleoacidic proteins (NAP) from combined 4-7 M-guanidine hydrochloride protein extracts showed saturable binding by [3H]-oestradiol receptors, whereas binding to hen DNA did not saturate. The high affinity, low capacity, and specificity of binding of oestrogen receptors to NAP was similar to that found in intact chromatin. Thus, chromatin acceptor proteins for the oestrogen receptor have been partially isolated and characterized in the hen oviduct and display properties similar to that reported for the acceptor proteins of the progesterone receptor.


RNA interference of gonadotropin-inhibitory hormone gene induces arousal in songbirds.
Gonadotropin-inhibitory hormone (GnIH) was originally identified in quail as a hypothalamic neuropeptide inhibitor of pituitary gonadotropin synthesis and release. However, GnIH neuronal fibers do not only terminate in the median eminence to control anterior pituitary function but also extend widely in the brain, suggesting it has multiple roles in the regulation of behavior. To identify the role of GnIH neurons in the regulation of behavior, we investigated the effect of RNA interference (RNAi) of the GnIH gene on the behavior of white-crowned sparrows, a highly social songbird species. Administration of small interfering RNA against GnIH precursor mRNA into the third ventricle of male and female birds reduced resting time, spontaneous production of complex vocalizations, and stimulated brief agonistic vocalizations. GnIH RNAi further enhanced song production of short duration in male birds when they were challenged by playbacks of novel male songs. These behaviors resembled those of breeding birds during territorial defense. The overall results suggest that GnIH gene silencing induces arousal. In addition, the activities of male and female birds were negatively correlated with GnIH mRNA expression in the paraventricular nucleus. Density of GnIH neuronal fibers in the ventral tegmental area was decreased by GnIH RNAi treatment in female birds, and the number of gonadotropin-releasing hormone neurons that received close appositions of GnIH neuronal fiber terminals was negatively correlated with the activity of male birds. In summary, GnIH may decrease arousal level resulting in the inhibition of specific motivated behavior such as in reproductive contexts.


Gender Differences in Factors Related to HIV Risk Behaviors among People Who Inject Drugs in North-East India.
People who inject drugs (PWID) in India are at high risk for HIV, with women being at elevated risk. Using a socio-ecological framework, this study assessed whether factors associated with HIV transmission risk behaviors differed across men and women PWID. Data for this cross-sectional study were collected from 6449 PWID in 7 cities in Northeast India. Men (n = 5653) and women (n = 796) PWID were recruited using respondent-driven sampling (RDS). We assessed sex differences in two recent HIV transmission risk behaviors: multiple sex partners and needle/syringe sharing. We used multi-level logistic regression models, which incorporated sampling weights and random intercepts for city, to assess factors associated with these HIV risks, separately among men and women. The prevalence of HIV was significantly higher among women than men (53% vs 18.4%, p<0.01). Nearly 13% of men and 8% of women (p = .30) had multiple partners. Employment in men and relationship status and stigma in women were significantly associated with multiple partners. Approximately 25% of men and 19% of women engaged in needle sharing (p = .16). Younger age in women and depression symptoms in men were significantly associated with increased risk for sharing needles. We found that sexual and drug related risk behaviors were common among PWID in Northeast India, and there were differences between men and women in the socio-ecologic correlates of these behaviors. Contextually-integrated and gender-specific HIV prevention and intervention efforts are needed that consider factors at individual, interpersonal- and community-levels that uniquely impact HIV risks among PWID.


Distinct modes of floor plate induction in the chick embryo.
To begin to reconcile models of floor plate formation in the vertebrate neural tube, we have performed experiments aimed at understanding the development of the early floor plate in the chick embryo. Using real-time analyses of cell behaviour, we provide evidence that the principal contributor to the early neural midline, the future anterior floor plate, exists as a separate population of floor plate precursor cells in the epiblast of the gastrula stage embryo, and does not share a lineage with axial mesoderm. Analysis of the tissue interactions associated with differentiation of these cells to a floor plate fate reveals a role for the nascent prechordal mesoderm, indicating that more than one inductive event is associated with floor plate formation along the length of the neuraxis. We show that Nr1, a chick nodal homologue, is expressed in the nascent prechordal mesoderm and we provide evidence that Nodal signalling can cooperate with Shh to induce the epiblast precursors to a floor-plate fate. These results indicate that a shared lineage with axial mesoderm cells is not a pre-requisite for floor plate differentiation and suggest parallels between the development of the floor plate in amniote and anamniote embryos.


3' non-translated sequences in Drosophila cyclin B transcripts direct posterior pole accumulation late in oogenesis and peri-nuclear association in syncytial embryos.
We have characterised forms of the Drosophila cyclin B transcript that differ as a result of a splicing event which removes a nucleotide segment from the 3' untranslated region. In oogenesis, both cyclin A RNA and a shorter form of the cyclin B transcript are seen in the cells of the germarium that are undergoing mitosis. The shorter cyclin B transcript alone is then detectable in the presumptive oocyte until stages 7-8 of oogenesis. Both cyclin A RNA and a longer form of the cyclin B RNA are then synthesised in the nurse cells during stages 9-11, to be deposited in the oocyte during stages 11-12. These transcripts become evenly distributed throughout the oocyte cytoplasm but, in addition, those of cyclin B become concentrated at the posterior pole. Examination of the distributions of RNAs transcribed from chimeric cyclin genes indicates that sequences in the 3' untranslated region of the larger cyclin B RNA are required both for it to become concentrated at the posterior pole and to direct those transcripts in the body of the syncytial embryo to their peri-nuclear localisation. These sequences are disrupted by the splicing event which generates smaller cyclin B transcripts.


The mother superior mutation ablates foxd3 activity in neural crest progenitor cells and depletes neural crest derivatives in zebrafish.
The zebrafish mutation mother superior (mosm188) leads to a depletion of neural crest (NC) derivatives including the craniofacial cartilage skeleton, the peripheral nervous system (sympathetic neurons, dorsal root ganglia, enteric neurons), and pigment cells. The loss of derivatives is preceded by a reduction in NC-expressed transcription factors, snail1b, sox9b, sox10, and a specific loss of foxd3 expression in NC progenitor cells. We employed genetic linkage analysis and physical mapping to place the mosm188 mutation on zebrafish chromosome 6 in the vicinity of the foxd3 gene. Furthermore, we found that mosm188 does not complement the sym1/foxd3 mutation, indicating that mosm188 resides within the foxd3 locus. Injection of PAC clones containing the foxd3 gene into mosm188 embryos restored foxd3 expression in NC progenitors and suppressed the mosm188 phenotype. However, sequencing the foxd3 transcribed area in mosm188 embryos did not reveal nucleotide changes segregating with the mosm188 phenotype, implying that the mutation most likely resides outside the foxd3-coding region. Based on these findings, we propose that the mosm188 mutation perturbs a NC-specific foxd3 regulatory element. Further analysis of mosm188 mutants and foxd3 morphants revealed that NC cells are initially formed, suggesting that foxd3 function is required to maintain the pool of NC progenitors.


Phase 1/2 study of rilotumumab (AMG 102), a hepatocyte growth factor inhibitor, and erlotinib in patients with advanced non-small cell lung cancer.
Activation of the mesenchymal-epidermal transition factor (MET) tyrosine kinase and its ligand, hepatocyte growth factor (HGF), is implicated in resistance to epidermal growth factor receptor (EGFR) inhibitors. In this phase 1/2 trial, rilotumumab (an anti-HGF antibody) combined with erlotinib was evaluated in patients with metastatic, previously treated non-small cell lung cancer. In phase 1, a dose de-escalation design was adopted with rilotumumab starting at 15 mg/kg intravenously every 3 weeks and oral erlotinib 150 mg daily. In phase 2, the disease control rate (DCR) (according to Response Evaluation Criteria in Solid Tumors) of the combination was evaluated using a Simon 2-stage design. The biomarkers examined included 10 plasma-circulating molecules associated with the EGFR and MET pathways. Without indications for de-escalation, the recommended phase 2 dose was dose level 0. Overall, 45 response-evaluable patients were enrolled (13 with squamous carcinoma, 32 with adenocarcinoma; 2 had confirmed EGFR mutations, 33 had confirmed wild-type [WT] EGFR, and 7 had KRAS mutations). The DCR for all patients was 60% (90% confidence interval [CI], 47.1%-71.3%). Median progression-free survival was 2.6 months (90% CI, 1.4-2.7 months), and median overall survival was 6.6 months (90% CI, 5.6-8.9 months). Among patients with WT EGFR, the DCR was 60.6% (90% CI, 46.3%-73.3%), median progression-free survival was 2.6 months (90% CI, 1.4-2.7 months), and median overall survival was 7.0 months (90% CI, 5.6-13.4 months). Elevated baseline levels of neuregulin 1 were associated with longer progression-free survival (hazard ratio, 0.41; 95% CI, 0.19-0.87), whereas elevated amphiregulin levels were associated with more rapid progression (hazard ratio, 2.14; 95% CI, 1.48-3.08). Combined rilotumumab and erlotinib had an acceptable safety profile, and the DCR met the prespecified criteria for success. In the EGFR WT group, the DCR exceeded published reports for erlotinib alone. High circulating levels of neuregulin 1 may indicate sensitivity to this combination. Cancer 2017;123:2936-44. © 2017 American Cancer Society.


Carotid-ophthalmic aneurysms.
Thirty-two cases of carotid-ophthalmic aneurysms are reviewed. As with intracranial aneurysms in other positions they present mainly with subarachnoid haemorrhage but, in spite of their close proximity to the optic nerve, visual involvement is infrequent. They are more common in women, more frequent on the left side, and more prone to multiplicity. In cases of multiple aneurysms a carotid-ophthalmic aneurysm is usually an incidental finding. Detailed angiographic studies employing various projections are required before treatment can be planned. Yet angiography does not always disclose some of the technical difficulties that may be encountered during surgery. Different methods of treating these aneurysms are discussed and suggestions for safe direct surgery made.


Autoreactive helper T cells alleviate the need for intrinsic TLR signaling in autoreactive B cell activation.
T cells play a significant role in the pathogenesis of systemic autoimmune diseases, including systemic lupus erythematosus; however, there is relatively little information on the nature and specificity of autoreactive T cells. Identifying such cells has been technically difficult because they are likely to be rare and low affinity. Here, we report a method for identifying autoreactive T cell clones that recognize proteins contained in autoantibody immune complexes, providing direct evidence that functional autoreactive helper T cells exist in the periphery of normal mice. These T cells significantly enhanced autoreactive B cell proliferation and altered B cell differentiation in vivo. Most importantly, these autoreactive T cells were able to rescue many aspects of the TLR-deficient AM14 (anti-IgG2a rheumatoid factor) B cell response, suggesting that TLR requirements can be bypassed. This result has implications for the efficacy of TLR-targeted therapy in the treatment of ongoing disease.


How do high preoperative pain scores impact the clinical course and outcomes for patients undergoing lumbar microdiscectomy?
Preoperative pain assessment is often used to gauge the amount of disability in patients with lumbar disc herniation. How high preoperative pain scores impact the clinical course and outcomes of patients after lumbar microdiscectomy is not always clear. Here, the authors aimed to determine whether patients reporting higher preoperative pain scores have worse outcomes after lumbar microdiscectomy than those reporting lower preoperative scores. The authors performed a retrospective review of patients with symptomatic lumbar disc herniations that had failed to improve with nonsurgical methods and who had undergone tubular lumbar microdiscectomy. Health-related quality of life (HRQOL) scores had been collected in the preoperative and postoperative period. The anatomical severity of disease was graded based on lumbar disc health (Pfirrmann classification), facet degeneration, thecal sac cross-sectional area, and disc herniation grade. Data on each patient's narcotic consumption and length of stay were collected. A Student t-test and chi-square test were used to compare patients with high preoperative pain scores (HP cohort) and those with lower preoperative scores (non-HP cohort). One hundred thirty-eight patients were included in this analysis. The 47 patients in the HP cohort had taken more preoperative opioids (12.0 ± 21.2 vs 3.6 ± 11.1 morphine equivalent doses, p = 0.01). However, there was no statistically significant difference in Pfirrmann classification (p > 0.15), facet grade (p > 0.11), thecal sac cross-sectional area (p = 0.45), or disc herniation grade (p = 0.39) between the HP and non-HP cohorts. The latter cohort had statistically significant higher preoperative PROMIS scores (36.5 ± 7.0 vs 29.9 ± 6.4, p < 0.001), SF-12 mental component summary scores (48.7 ± 11.5 vs 38.9 ± 16.1, p < 0.001), and SF-12 physical component summary scores (PCS; 32.4 ± 8.6 vs 27.5 ± 10.0, p = 0.005), but a lower Oswestry Disability Index (56.4 ± 22.1 vs 35.4 ± 15.5, p < 0.001). There were only two time points after microdiscectomy when the HP cohort had worse HRQOL scores: at the 2-week follow-up for SF-12 PCS scores (32.4 ± 8.6 vs 29.3 ± 7.1, p = 0.03) and the 12-week follow-up for PROMIS scores (45.2 ± 9.5 vs 39.5 ± 7.1, p = 0.01). All other postoperative HRQOL measurements were similar between the two cohorts (p > 0.05). A patient's perceived severity of disease often does not correlate with the actual clinical pathology on imaging. Although patients who report high pain and have a symptomatic lumbar disc herniation may describe their pain as more extreme, they should be counseled that the outcomes of microdiscectomy are positive.


Arginine/lysine-rich nuclear localization signals mediate interactions between dimeric STATs and importin alpha 5.
Interferon stimulation results in tyrosine phosphorylation, dimerization, and nuclear import of STATs (signal transducers and activators of transcription). Proteins to be targeted into the nucleus usually contain nuclear localization signals (NLSs), which interact with importin alpha. Importin alpha binds to importin beta, which docks the protein complex to nuclear pores, and the complex translocates into the nucleus. Here we show that baculovirus-produced and -activated STAT1 homodimers and STAT1-STAT2 heterodimers directly interacted with importin alpha 5 (NPI-1). This interaction was very stable and was dependent on lysines 410 and 413 of STAT1. Only STAT dimers that had two intact NLS elements, one in each monomer, were able to bind to importin alpha 5. STAT-importin alpha 5 complexes apparently consisted of two STAT and two importin alpha molecules. STAT NLS-dependent colocalization of importin alpha 5 with STAT1 or STAT2 was seen in the nucleus of transfected cells. gamma-Activated sequence DNA elements efficiently inhibited STAT binding to importin alpha 5 suggesting that the DNA and importin alpha binding sites are close to each other in STAT dimers. Our results demonstrate that specific NLSs in STATs mediate direct interactions of STAT dimers with importin alpha, which activates the nuclear import process.


A Phase II study of a histamine H₃ receptor antagonist GSK239512 for cognitive impairment in stable schizophrenia subjects on antipsychotic therapy.
This Phase II exploratory study assessed GSK239512, a brain penetrant histamine H₃ receptor antagonist, versus placebo on cognitive impairment in 50 stable outpatients with schizophrenia. Subjects were randomized to placebo or GSK239512 for 7 weeks (4 weeks titration). GSK239512 was associated with a small positive effect size (ES) on the CogState Schizophrenia Battery (CSSB) Composite Score (ES=0.29, CI=-0.40, 0.99) relative to placebo (primary endpoint). GSK239512's ES on CSSB domains were generally positive or neutral except Processing Speed, which favored placebo (ES=-0.46). Effects on the MATRICS Consensus Cognitive Battery were mostly neutral or favored placebo. GSK239512 was generally well tolerated with an adverse event profile consistent with the known class pharmacology. There was no evidence of overall beneficial effects of GSK239512 for CIAS in this population.


Conditional neuronal nitric oxide synthase overexpression impairs myocardial contractility.
The role of the neuronal NO synthase (nNOS or NOS1) enzyme in the control of cardiac function still remains unclear. Results from nNOS(-/-) mice or from pharmacological inhibition of nNOS are contradictory and do not pay tribute to the fact that probably spatial confinement of the nNOS enzyme is of major importance. We hypothesize that the close proximity of nNOS and certain effector molecules like L-type Ca(2+)-channels has an impact on myocardial contractility. To test this, we generated a new transgenic mouse model allowing conditional, myocardial specific nNOS overexpression. Western blot analysis of transgenic nNOS overexpression showed a 6-fold increase in nNOS protein expression compared with noninduced littermates (n=12; P<0.01). Measuring of total NOS activity by conversion of [(3)H]-l-arginine to [(3)H]-l-citrulline showed a 30% increase in nNOS overexpressing mice (n=18; P<0.05). After a 2 week induction, nNOS overexpression mice showed reduced myocardial contractility. In vivo examinations of the nNOS overexpressing mice revealed a 17+/-3% decrease of +dp/dt(max) compared with noninduced mice (P<0.05). Likewise, ejection fraction was reduced significantly (42% versus 65%; n=15; P<0.05). Interestingly, coimmunoprecipitation experiments indicated interaction of nNOS with SR Ca(2+)ATPase and additionally with L-type Ca(2+)- channels in nNOS overexpressing animals. Accordingly, in adult isolated cardiac myocytes, I(Ca,L) density was significantly decreased in the nNOS overexpressing cells. Intracellular Ca(2+)-transients and fractional shortening in cardiomyocytes were also clearly impaired in nNOS overexpressing mice versus noninduced littermates. In conclusion, conditional myocardial specific overexpression of nNOS in a transgenic animal model reduced myocardial contractility. We suggest that nNOS might suppress the function of L-type Ca(2+)-channels and in turn reduces Ca(2+)-transients which accounts for the negative inotropic effect.


Family load estimates and risk factors of anxiety disorders in a nationwide three generation study.
The present study investigated how often anxiety disorders with different ages of onset occurred in affected families compared to control families. Furthermore, the study addressed the impact of sex, region of residence, year and month of birth, and parental age at birth. The sample included N=1373 child and adolescent psychiatric participants born between 1952 and 2000 and registered in the Danish Psychiatric Central Register (DPCR) who developed an anxiety disorder before the age of 18. N =4019 controls without any psychiatric diagnosis before age 18, were matched for age, sex, and residential region. Psychiatric diagnoses were also obtained for parents, siblings, and offspring. A family load component was obtained by using various mixed regression models. Anxiety disorders occurred significantly more often in case than in control families. Having a mother, father, or a sibling with the disorder was proven to be a risk factor. Female sex, year of birth, and region of residence were also associated with having an anxiety disorder. Furthermore, case relatives did not develop an anxiety disorder earlier than control relatives. These findings, based on a very large and representative dataset, provide further and solid evidence for the family aggregation of anxiety disorders.


Long-term effectiveness of faecal immunochemical test screening for proximal and distal colorectal cancers.
To measure the effects of faecal immunochemical test (FIT) for colorectal cancer (CRC) screening on overall and site-specific long-term effectiveness of population-based organised service screening. A prospective cohort study of Taiwanese nationwide biennial FIT screening was performed. A total of 5 417 699 eligible subjects were invited to attend screening from 2004 through 2009 and were followed up until 2014. We estimated the adjusted relative rates (aRRs) on the effectiveness of reducing advanced-stage CRC (stage II+) and CRC death by Bayesian Poisson regression models with the full adjustment for a cascade of self-selection factors (including the screening rate and the colonoscopy rate) and the completeness of colonoscopy together with demographic features. FIT screening (exposed vs unexposed) reduced the incidence of advanced-stage CRC (48.4 vs 75.7 per 100 000) and mortality (20.3 vs 41.3 per 100 000). Statistically significant reductions of both incidence of advanced-stage CRCs (aRR=0.66, 95% CI 0.63 to 0.70) and deaths from CRC (aRR=0.60, 95% CI 0.57 to 0.64) were noted. FIT screening was more effective in reducing distal advanced-stage CRCs (aRR=0.61, 95% CI 0.58 to 0.64) and CRC mortality (aRR=0.56, 95% CI 0.53 to 0.69) than proximal advanced CRCs (aRR=0.84, 95% CI 0.77 to 0.92) and CRC mortality (aRR=0.72, 95% CI 0.66 to 0.80). A large-scale population-based biennial FIT screening demonstrates 34% significant reduction of advanced-stage CRCs and 40% reduction of death from CRC with larger long-term effectiveness in the distal colon than the proximal colon. Our findings provide a strong and consistent evidence-based policy for supporting a sustainable population-based FIT organised service screening worldwide. The disparity of site-specific long-term effectiveness also provides an insight into the remedy for lower effectiveness of FIT screening in the proximal colon.