Evaluating the Impact of a Point-of-Care Cardiometabolic Clinical Decision Support Tool on Clinical Efficiency Using Electronic Health Record Audit Log Data: Algorithm Development and Validation.
Electronic health record (EHR) systems are becoming increasingly complicated, leading to concerns about rising physician burnout, particularly for primary care physicians (PCPs). Managing the most common cardiometabolic chronic conditions by PCPs during a limited clinical time with a patient is challenging. This study aimed to evaluate a Cardiometabolic Sutter Health Advanced Reengineered Encounter (CM-SHARE), a web-based application to visualize key EHR data, on the EHR use efficiency. We developed algorithms to identify key clinic workflow measures (eg, total encounter time, total physician time in the examination room, and physician EHR time in the examination room) using audit data, and we validated and calibrated the measures with time-motion data. We used a pre-post parallel design to identify propensity score-matched CM-SHARE users (cases), nonusers (controls), and nested-matched patients. Cardiometabolic encounters from matched case and control patients were used for the workflow evaluation. Outcome measures were compared between the cases and controls. We applied this approach separately to both the CM-SHARE pilot and spread phases. Time-motion observation was conducted on 101 primary care encounters for 9 PCPs in 3 clinics. There was little difference (<0.8 minutes) between the audit data-derived workflow measures and the time-motion observation. Two key unobservable times from audit data, physician entry into and exiting the examination room, were imputed based on time-motion studies. CM-SHARE was launched with 6 pilot PCPs in April 2016. During the prestudy period (April 1, 2015, to April 1, 2016), 870 control patients with 2845 encounters were matched with 870 case patients and encounters, and 727 case patients with 852 encounters were matched with 727 control patients and 3754 encounters in the poststudy period (June 1, 2016, to June 30, 2017). Total encounter time was slightly shorter (mean -2.7, SD 1.4 minutes, 95% CI -4.7 to -0.9; mean -1.6, SD 1.1 minutes, 95% CI -3.2 to -0.1) for cases than controls for both periods. CM-SHARE saves physicians approximately 2 minutes EHR time in the examination room (mean -2.0, SD 1.3, 95% CI -3.4 to -0.9) compared with prestudy period and poststudy period controls (mean -1.9, SD 0.9, 95% CI -3.8 to -0.5). In the spread phase, 48 CM-SHARE spread PCPs were matched with 84 control PCPs and 1272 cases with 3412 control patients, having 1119 and 4240 encounters, respectively. A significant reduction in total encounter time for the CM-SHARE group was observed for short appointments (≤20 minutes; 5.3-minute reduction on average) only. Total physician EHR time was significantly reduced for both longer and shorter appointments (17%-33% reductions). Combining EHR audit log files and clinical information, our approach offers an innovative and scalable method and new measures that can be used to evaluate clinical EHR efficiency of digital tools used in clinical settings.


Bioinspired Mineralization Using Chondrocyte Membrane Nanofragments.
Biomineralization involves complex processes and interactions between organic and inorganic matters, which are controlled in part by the cells. The objectives of this study were, first, to perform a systematic and ultrastructural investigation of the initial mineral formation during secondary ossification center of mouse femur based on material science and biology viewpoint, and then develop novel biomaterials for mineralization based on the in vivo findings. First, we identified the very initial mineral deposition at postnatal day 5 (P5) at the medial side of femur epiphysis by nanocomputed tomography. Initial minerals were found in the surroundings of hypertrophic chondrocytes. Interestingly, histological and immunohistochemical analyses showed that initial mineralization until P6 was based on chondrocyte activity only, i.e., it occurred in the absence of osteoblasts. Moreover, electron microscopy-based ultrastructural analysis showed that cell-secreted matrix vesicles were absent in the early steps of osteoblast-independent endochondral ossification. Instead, chondrocyte membrane nanofragments were found in the fibrous matrix surrounding the hypertrophic chondrocytes. EDS analysis and electron diffraction study indicated that cell membrane nanofragments were not mineralized material, and could be the nucleation site for the newly formed calcospherites. The phospholipids in the cell membrane nanofragments could be a source of phosphate for subsequent calcium phosphate formation, which initially was amorphous, and later transformed into apatite crystals. Finally, artificial cell nanofragments were synthesized from ATDC5 chondrogenic cells, and in vitro assays showed that these nanofragments could promote mineral formation. Taken together, these results indicated that cell membrane nanofragments were the nucleation site for mineral formation, and could potentially be used as material for manipulation of biomineralization.


Modification of planned actions.
To elucidate the time course and processes underlying pre-movement modification of planned actions, participants prepared to make an action at a time of their own choosing within a specified temporal window. In some conditions, participants prepared to make a single right index finger key press, whereas in others, they prepared to make a sequence of two key presses consisting of a right index finger key press followed by a right middle finger key press. On a proportion of trials, their internal preparation was interrupted by an auditory tone, in response to which they made either: the same action as they were intending, a different action requiring an additional effector (i.e. switch from preparing a single right index finger key press to executing a right index, middle finger sequence), or a different action requiring one less effector (i.e. switch from preparing a right index, middle finger sequence to executing a right index finger key press). For unmodified actions, switching from an internally generated to an externally triggered mode of response production produced a significant reaction time cost (RT cost) for both single and sequential actions, with the cost for single actions being significantly greater than that for sequential actions. Given that the RT cost did not increase as the complexity of the actions increased it is unlikely that the source of the cost is related to motor execution processes, and it is suggested that it may arise at a higher level cognitive stage of processing. In addition, reaction times to produce modified actions were significantly greater than those to produce unmodified actions. Finally, it took significantly longer to produce modified actions requiring one less effector than to produce modified actions requiring one more effector. We suggest that two time-consuming processes are involved in switching between internally generated and externally triggered actions that are modified or unmodified: a trigger switch cost when the same action has to be produced in response to an external trigger as opposed to an internal trigger, and a switch cost reflecting changes in the pattern of executed motor commands when modification is necessary. It is suggested that such processes may be mediated by regions of the frontal lobes.


Determinants and prevalence of e-cigarette use throughout the European Union: a secondary analysis of 26 566 youth and adults from 27 Countries.
This study assessed the prevalence and determinants of e-cigarette use among persons aged ≥15 years in 27 European Union (EU) member countries during 2012. The 2012 Eurobarometer 385 (77.1) survey was analysed for n=26 566 respondents. Knowledge, perception of harm, and determinants of e-cigarettes use were assessed, while separate regression analyses among current (n=7352) and former cigarette smokers (n=5782) were performed. National estimates of the number of e-cigarette users were also extrapolated. 20.3% of current smokers, 4.7% of ex-smokers, and 1.2% of never cigarette smokers in the EU reported having ever used an e-cigarette (overall approximately 29.3 million adults). Among smokers, ever e-cigarette use was more likely among 15-24-year-olds (aOR 3.13, 95% CI 2.22 to 4.54) and 25-39-year-olds (aOR 2.00, 95% CI 1.47 to 2.78) in comparison to older smokers, and among those who smoked 6-10 cigarettes/day (aOR 1.53, 95% CI 1.10 to 2.13) or 11-20 cigarettes/day (aOR 2.07, 95% CI 1.52 to 2.81) in comparison to very light smokers (≤5 cigarettes/day). Moreover, e-cigarette use was more likely among smokers who had made a past year quit attempt (aOR 2.08, 95% CI 1.67 to 2.58). E-cigarette use among ex-smokers was associated only with the respondents' age, with younger ex-smokers being more likely to have ever used an e-cigarette. A substantial number of EU adults have ever used e-cigarettes. Ever users were more likely to be younger, current smokers, or past-year quit attempters. These findings underscore the need to evaluate the potential long term impact of e-cigarette use on consumer health, cessation and nicotine addiction and formulate a European framework for e-cigarette regulation within the revised EU Tobacco Product Directive.


The Neandertals of northeastern Iberia: new remains from the Cova del Gegant (Sitges, Barcelona).
The present study describes a new juvenile hominin mandible and teeth and a new juvenile humerus from level V of the GP2 gallery of Cova del Gegant (Spain). The mandible (Gegant-5) preserves a portion of the right mandibular corpus from the M1 distally to the socket for the dc mesially, and the age at death is estimated as 4.5-5.0 years. Gegant-5 shows a single mental foramen located under the dm1/dm2 interdental septum, a relatively posterior placement compared with recent hominins of a similar developmental age. The mental foramen in Gegant-5 is also placed within the lower half of the mandibular corpus, as in the previously described late adolescent/adult mandible (Gegant-1) from this same Middle Paleolithic site. The Gegant-5 canine shows pronounced marginal ridges, a distal accessory ridge, and a pronounced distolingual tubercle. The P3 shows a lingually-displaced protoconid cusp tip and a distal accessory ridge. The P4 shows a slightly asymmetrical crown outline, a continuous transverse crest, a mesially placed metaconid cusp tip, a slight distal accessory ridge, and an accessory lingual cusp. The M1 shows a Y5 pattern of cusp contact and a well-developed and deep anterior fovea bounded posteriorly by a continuous midtrigonid crest. Gegant-4 is the distal portion of a left humerus from a juvenile estimated to be between 5 and 7 years old at death. The specimen shows thick cortical bone. Although fragmentary, the constellation of morphological and metric features indicates Neandertal affinities for these specimens. Their spatial proximity at the site and similar ages at death suggest these remains may represent a single individual. The addition of these new specimens brings the total number of Neandertal remains from the Cova del Gegant to five, and this site documents the clearest evidence for Neandertal fossils associated with Middle Paleolithic stone tools in this region of the Iberian Peninsula.


Intrinsically disordered regions regulate both catalytic and non-catalytic activities of the MutLα mismatch repair complex.
Intrinsically disordered regions (IDRs) are present in at least 30% of the eukaryotic proteome and are enriched in chromatin-associated proteins. Using a combination of genetics, biochemistry and single-molecule biophysics, we characterize how IDRs regulate the functions of the yeast MutLα (Mlh1-Pms1) mismatch repair (MMR) complex. Shortening or scrambling the IDRs in both subunits ablates MMR in vivo. Mlh1-Pms1 complexes with shorter IDRs that disrupt MMR retain wild-type DNA binding affinity but are impaired for diffusion on both naked and nucleosome-coated DNA. Moreover, the IDRs also regulate the adenosine triphosphate hydrolysis and nuclease activities that are encoded in the structured N- and C-terminal domains of the complex. This combination of phenotypes underlies the catastrophic MMR defect seen with the mutant MutLα in vivo. More broadly, this work highlights an unanticipated multi-functional role for IDRs in regulating both facilitated diffusion on chromatin and nucleolytic processing of a DNA substrate.


UFP-101 antagonizes the spinal antinociceptive effects of nociceptin/orphanin FQ: behavioral and electrophysiological studies in mice.
Nociceptin/orphanin FQ (N/OFQ) modulates various biological functions, including nociception, via selective stimulation of the N/OFQ peptide receptor (NOP). Here we used the NOP selective antagonist UFP-101 to characterize the receptor involved in the spinal antinociceptive effects of N/OFQ evaluated in the mouse tail withdrawal assay and to investigate the mechanism underlying this action by assessing excitatory postsynaptic currents (EPSC) in laminas I and II of the mouse spinal cord dorsal horn with patch-clamp techniques. Intrathecal (i.t.) injection of N/OFQ in the range of 0.1-10 nmol produced a dose dependent antinociceptive effect, which was prevented by UFP-101, but not by naloxone. In contrast the antinociceptive effect of the mu-opioid peptide receptor agonist endomorphin-1 was blocked by naloxone but not by UFP-101. Moreover, N/OFQ and endomorphin-1 induced a significant antinociceptive effect in wild type mice while in mice knockout for the NOP receptor gene only endomorphin-1 was found to be active. In mouse spinal cord slices 1 microM N/OFQ reduced EPSC to 60+/-4% of control values. This inhibitory effect was reversed in a concentration dependent manner by UFP-101 (pA2 value 6.44). The present results demonstrate that N/OFQ-induced spinal antinociception in vivo and inhibition of spinal excitatory transmission in vitro are mediated by receptors of the NOP type.


A novel environmental chamber for neuronal network multisite recordings.
Environmental stability is a critical issue for neuronal networks in vitro. Hence, the ability to control the physical and chemical environment of cell cultures during electrophysiological measurements is an important requirement in the experimental design. In this work, we describe the development and the experimental verification of a closed chamber for multisite electrophysiology and optical monitoring. The chamber provides stable temperature, pH and humidity and guarantees cell viability comparable to standard incubators. Besides, it integrates the electronics for long-term neuronal activity recording. The system is portable and adaptable for multiple network housings, which allows performing parallel experiments in the same environment. Our results show that this device can be a solution for long-term electrophysiology, for dual network experiments and for coupled optical and electrical measurements.


Can marijuana make it better? Prospective effects of marijuana and temperament on risk for anxiety and depression.
Increases in marijuana use in recent years highlight the importance of understanding how marijuana affects mental health. Of particular relevance is the effect of marijuana use on anxiety and depression given that marijuana use is highest among late adolescents/early adults, the same age range in which risk for anxiety and depression is the highest. Here we examine how marijuana use moderates the effects of temperament on level of anxiety and depression in a prospective design in which baseline marijuana use and temperament predict anxiety and depression 1 year later. We found that harm avoidance (HA) is associated with higher anxiety and depression a year later, but only among those low in marijuana use. Those higher in marijuana use show no relation between HA and symptoms of anxiety and depression. Marijuana use also moderated the effect of novelty seeking (NS), with symptoms of anxiety and depression increasing with NS only among those with high marijuana use. NS was unrelated to symptoms of anxiety and depression among those low in marijuana use. The temperament dimension of reward dependence was unrelated to anxiety and depression symptoms. Our results suggest that marijuana use does not have an invariant relationship with anxiety and depression, and that the effects of relatively stable temperament dimensions can be moderated by other contextual factors.


A concept analysis of renal supportive care: the changing world of nephrology.
This paper is a report of a concept analysis of renal supportive care. Approximately 1.5 million people worldwide are kept alive by renal dialysis. As services are required to support patients who decide not to start or to withdraw from dialysis, the term renal supportive care is emerging. Being similar to the terms palliative care, end-of-life care, terminal care and conservative management, there is a need for conceptual clarity. Rodgers' evolutionary method was used as the organizing framework for this concept analysis. Data were collected from a review of CINAHL, Medline, PsycINFO, British Nursing Index, International Bibliography of the Social Sciences and ASSIA (1806-2006) using, 'renal' and 'supportive care' as keywords. All articles with an abstract were considered. The World Wide Web was also searched in English utilizing the phrase 'renal supportive care'. Five attributes of renal supportive care were identified: available from diagnosis to death with an emphasis on honesty regarding prognosis and impact of disease; interdisciplinary approach to care; restorative care; family and carer support and effective, lucid communication to ensure informed choice and clear lines of decision-making. Renal supportive care is a dynamic and emerging concept relevant, but not limited to, the end phase of life. It suggests a central philosophy underpinning renal service development that allows patients, carers and the multidisciplinary team time to work together to realize complex goals. It has relevance for the renal community and is likely to be integrated increasingly into everyday nephrology practice.


Simvastatin enhances bone morphogenetic protein receptor type II expression.
Statins confer therapeutic benefits in systemic and pulmonary vascular diseases. Bone morphogenetic protein (BMP) receptors serve essential signaling functions in cardiovascular development and skeletal morphogenesis. Mutations in BMP receptor type II (BMPR2) are associated with human familial and idiopathic pulmonary arterial hypertension, and pathologic neointimal proliferation of vascular endothelial and smooth muscle cells within small pulmonary arteries. In severe experimental pulmonary hypertension, simvastatin reversed disease and conferred a 100% survival advantage. Here, modulation of BMPR2 gene expression by simvastatin is characterized in human embryonic kidney (HEK) 293T, pulmonary artery smooth muscle, and lung microvascular endothelial cells (HLMVECs). A 1.4kb BMPR2 promoter containing Egr-1 binding sites confers reporter gene activation in 293T cells which is partially inhibited by simvastatin. Simvastatin enhances steady-state BMPR2 mRNA and protein expression in HLMVEC, through posttranscriptional mRNA stabilization. Simvastatin induction of BMPR2 expression may improve BMP-BMPR2 signaling thereby enhancing endothelial differentiation and function.


Roles for cysteine residues in the regulatory CXXC motif of human mitochondrial branched chain aminotransferase enzyme.
The redox-active dithiol/disulfide C315-Xaa-Xaa-C318 center has been implicated in the regulation of the human mitochondrial branched chain aminotransferase isozyme (hBCATm) [Conway, M. E., Yennawar, N., Wallin, R., Poole, L. B., and Hutson, S. M. (2002) Biochemistry 41, 9070-9078]. To explore further the mechanistic details of this CXXC center, mutants of the Cys residues at positions 315 and 318 of hBCATm were individually and in combination converted to alanine or serine by site-directed mutagenesis (C315A, C315S, C318A, C318S, C315/318A, and C315/318S). The effects of these mutations on cofactor absorbance, secondary structures, steady-state kinetics, and sensitivity toward hydrogen peroxide (H(2)O(2)) treatment were examined. Neither the UV-visible spectroscopic studies nor the circular dichroism data showed any major perturbations in the structure of the mutants. Kinetic analyses of the CXXC mutant proteins indicated primarily a modest reduction in k(cat) with no significant change in K(m). The largest effect on the steady-state kinetics was observed with the C315 single mutants, in which substitution of the thiol group resulted in a reduced k(cat) (to 26-33% of that of wild-type hBCATm). Moreover, the C315 single mutants lost their sensitivity to oxidation by H(2)O(2). The kinetic parameters of the C318 mutants were largely unaffected by the substitutions, and as with wild-type hBCATm, reaction of the C318A mutant protein with H(2)O(2) resulted in the complete loss of activity. In the case of oxidized C318A, this loss was largely irreversible on incubation with dithiothreitol. Mass spectrometry and dimedone modification results revealed overoxidation of the thiol group at position 315 to sulfonic acid occurring via a sulfenic acid intermediate in the H(2)O(2)-treated C318A enzyme. Thus, C315 appears to be the sensor for redox regulation of BCAT activity, whereas C318 acts as the "resolving cysteine", allowing for reversible formation of a disulfide bond.


Shrinkage effect in ancestral maximum likelihood.
Ancestral maximum likelihood (AML) is a method that simultaneously reconstructs a phylogenetic tree and ancestral sequences from extant data (sequences at the leaves). The tree and ancestral sequences maximize the probability of observing the given data under a Markov model of sequence evolution, in which branch lengths are also optimized but constrained to take the same value on any edge across all sequence sites. AML differs from the more usual form of maximum likelihood (ML) in phylogenetics because ML averages over all possible ancestral sequences. ML has long been know to be statistically consistent--that is, it converges on the correct tree with probability approaching 1 as the sequence length grows. However, the statistical consistency of AML has not been formally determined, despite informal remarks in a literature that dates back 20 years. In this short note we prove a general result that implies that AML is statistically inconsistent. In particular we show that AML can 'shrink' short edges in a tree, resulting in a tree that has no internal resolution as the sequence length grows. Our results apply to any number of taxa.


Prevalence of Helicobacter pylori in HIV-infected, HAART-naïve Ugandan children: a hospital-based survey.
The aim of this survey was to determine the prevalence of and factors associated with Helicobacter pylori (H. pylori) colonization in HIV-infected, highly active antiretroviral therapy-naïve Ugandan children aged 0-12 years. In a hospital-based survey, 236 HIV-infected children were tested for H. pylori colonization using a faecal antigen test. A standardized interview with socio-demographic information and medical history was used to assess risk factors. A cluster of differentiation 4 (CD4) cell percentage was prevalent in most children. The overall prevalence of H. pylori in the HIV-infected children was 22.5%. Age-specific prevalence was as follows: up to one year, 14.7%; 1-3 years, 30.9%; and 3-12 years, 20.7%. HIV-infected children who were more seriously affected by their disease (low CD4 cell percentage or WHO clinical stage II-IV) were less likely to be colonized with H. pylori. There was a trend for a lower prevalence of H. pylori in children who had taken antibiotics for the preceding two weeks (21.6%) than in those who had not taken antibiotics (35.7%). There was no statistically significant difference in prevalence by gender, housing, congested living, education of the female caretaker, drinking water or toilet facilities. HIV-infected, HAART-naïve Ugandan children had a lower prevalence of H. pylori colonization compared with apparently healthy Ugandan children (44.3%). Children with a low CD4 cell percentage and an advanced clinical stage of HIV had an even lower risk of H. pylori colonization. Treatment with antibiotics due to co-morbidity with infectious diseases is a possible explanation for the relatively low prevalence.


Effects in feline gastrocnemius-soleus motoneurones induced by muscle fatigue.
Responses of gastrocnemius-soleus (G-S) motoneurones to stretches of the homonymous muscles were recorded intracellularly in decerebrate cats before, during and after fatiguing stimulation (FST) of G-S muscles. Ventral roots (VR) L7 and S1 were cut, and FST was applied to VR S1, a single FST session including 4 to 5 repetitions of 12-s periods of regular 40 s(-1) stimulation. Muscle stretches consisted of several phases of slow sinusoidal shortening-lengthening cycles and intermediate constant lengths. The maximal stretch of the muscles was 8.8 mm above the rest length. Effects of FST on excitatory postsynaptic potentials (EPSPs) and spikes evoked by the muscle stretches were studied in 12 motoneurones from ten experiments. Stretch-evoked EPSPs and firing were predominantly suppressed after FST, with the exception of a post-contraction increase of the first EPSP after FST, which was most likely due to after-effects in the activity of muscle spindle afferents. The post-fatigue suppression of EPSPs and spike activity was followed by restoration within 60-100 s. Additional bouts of FST augmented the intensity of post-fatigue suppression of EPSPs, with the spike activity sometimes disappearing completely. FST itself elicited EPSPs at latencies suggesting activation of muscle spindle group Ia afferents via stimulation of beta-fibres. The suppression of the stretch-evoked responses most likely resulted from fatigue-evoked activity of group III and IV muscle afferents. Presynaptic inhibition could be one of the mechanisms involved, but homosynaptic depression in the FST-activated group Ia afferents may also have contributed.


Phenotypic profiling of the human genome by time-lapse microscopy reveals cell division genes.
Despite our rapidly growing knowledge about the human genome, we do not know all of the genes required for some of the most basic functions of life. To start to fill this gap we developed a high-throughput phenotypic screening platform combining potent gene silencing by RNA interference, time-lapse microscopy and computational image processing. We carried out a genome-wide phenotypic profiling of each of the approximately 21,000 human protein-coding genes by two-day live imaging of fluorescently labelled chromosomes. Phenotypes were scored quantitatively by computational image processing, which allowed us to identify hundreds of human genes involved in diverse biological functions including cell division, migration and survival. As part of the Mitocheck consortium, this study provides an in-depth analysis of cell division phenotypes and makes the entire high-content data set available as a resource to the community.


Host Correlates of Avian Influenza Virus Infection in Wild Waterfowl of the Sacramento Valley, California.
Avian influenza viruses (AIVs) are distributed globally in members of the family Anatidae (waterfowl), and significant disease may occur when these viruses infect commercial poultry or humans. Early detection of AIV through surveillance of wild waterfowl is one measure to prevent future disease outbreaks. Surveillance efforts that are designed to account for host and environmental determinants of susceptibility to infection are likely to be most effective. However, these determinants have not been clearly delineated and may vary with location. Because some regions are at greater risk for AIV outbreaks, the factors that contribute to AIV infection of waterfowl in these areas are of interest. We investigated the prevalence of AIVs in hunter-killed waterfowl at wintering sites in California's Central Valley. Overall, AIV prevalence was 10.5% and, after controlling for age and sex, was greatest in northern shovelers (Spatula clypeata) and lowest in wood ducks (Aix sponsa). Overall, AIV prevalence was higher in females than in males, but this trend was driven by one sampling year and one waterfowl species (green-winged teal, Anas crecca). AIV prevalence in waterfowl was lower in samples collected from brackish wetlands compared with those collected from freshwater wetlands, suggesting that wetland type or other environmental factors contribute to AIV prevalence. This study adds to our understanding of the ecology of AIV infection in waterfowl and may assist in developing more efficient, targeted surveillance efforts for the detection of potentially harmful viruses circulating in North American waterfowl.


Internet-based behavioral activation and acceptance-based treatment for depression: a randomized controlled trial.
Internet-based cognitive behavior therapy for depression has been tested in several trials but there are no internet studies on behavioral activation (BA), and no studies on BA over the internet including components of acceptance and commitment therapy (ACT). The aim of this study was to develop and test the effects of internet-delivered BA combined with ACT against a waiting list control condition as a first test of the effects of treatment. Selection took place with a computerized screening interview and a subsequent semi-structured telephone interview. A total of 80 individuals from the general public were randomized to one of two conditions. The treatment lasted for 8 weeks after which both groups were assessed. We also included a 3 month follow-up. The treatment included interactive elements online and a CD-ROM for mindfulness and acceptance exercises. In addition, written support and feedback was given by a therapist every week. Results at posttreatment showed a large between group effect size on the Beck Depression inventory II d=0.98 (95%CI=0.51-1.44). In the treated group 25% (10/40) reached remission defined as a BDI score ≤ 10 vs. 5% (2/40) in the control group. Results on secondary measures were smaller. While few dropped out from the study (N=2) at posttreatment, the average number of completed modules was M=5.1 out of the seven modules. The study only included a waiting-list comparison and it is not possible to determine which treatment components were the most effective. We conclude that there is initial evidence that BA with components of ACT can be effective in reducing symptoms of depression.


Financial incentives for abstinence among socioeconomically disadvantaged individuals in smoking cessation treatment.
We evaluated the effectiveness of offering adjunctive financial incentives for abstinence (contingency management [CM]) within a safety net hospital smoking cessation program. We randomized participants (n = 146) from a Dallas County, Texas, Tobacco Cessation Clinic from 2011 to 2013 to usual care (UC; cessation program; n = 71) or CM (UC + 4 weeks of financial incentives; n = 75), and followed from 1 week before the quit date through 4 weeks after the quit date. A subset (n = 128) was asked to attend a visit 12 weeks after the scheduled quit date. Participants were primarily Black (62.3%) or White (28.1%) and female (57.5%). Most participants were uninsured (52.1%) and had an annual household income of less than $12 000 (55.5%). Abstinence rates were significantly higher for those assigned to CM than UC at all visits following the quit date (all Ps < .05). Point prevalence abstinence rates in the CM and UC groups were 49.3% versus 25.4% at 4 weeks after the quit date and 32.8% versus 14.1% at 12 weeks after the quit date. CM participants earned an average of $63.40 ($150 possible) for abstinence during the first 4 weeks after the scheduled quit date. Offering small financial incentives for abstinence might be an effective means to improve abstinence rates among socioeconomically disadvantaged individuals participating in smoking cessation treatment.


Establishing the germline in spiralian embyos.
Elucidating the origin of germ cells in embryos and larvae is often obscured by the fact that the typical germ cell markers vasa, nanos and piwi are not exclusively expressed in primordial germ cells (PGCs), but are also commonly found in undifferentiated somatic tissues and stem cells as part of an evolutionary conserved 'germline multipotency program' (Juliano et al., 2010). Hidden in the crowd of undifferentiated cells, the PGCs have occasionally been overlooked and their formation during early embryogenesis was only revealed recently by new methodological approaches (e.g. Wu et al., 2011). Spiralians are excellent model organisms to deepen our understanding of PGC formation, given the highly stereotypical cleavage that occurs during embryogenesis. In these species, detailed cell lineage studies enable the tracing of single cells up to gastrulation stages. Here, I review our knowledge of the origin of PGCs in these invertebrates. Similarities in PGC formation among spiralian phyla as well as peculiarities of the highly derived clitellates are discussed with respect to developmental mode and evolution. Furthermore, the issue of gonad regeneration in platyhelminths and the asexually reproducing oligochaete Enchytraeus japonensis is addressed. An alternative strategy of compensating for caudal regeneration is presented for the polychaete Platynereis dumerilli. Finally, the molecular bases of PGC specification and the question of germplasm are discussed.


Implementation of the Danish return-to-work program: process evaluation of a trial in 21 Danish municipalities.
The aim of this study was to evaluate the implementation of the Danish national return-to-work (RTW) program in 21 Danish municipalities. We conducted a structured process evaluation on (i) reach and recruitment, (ii) fidelity, (iii) dose-delivered, (iv) dose-received, and (v) context by formulating 29 implementation criteria and analyzing qualitative and quantitative data from administrative records, interviews, field notes, and questionnaires. All municipalities integrated the basic features of the RTW program into the existing framework of the sickness benefit management system to an acceptable degree, ie, establishment of RTW teams, participation of RTW team members in the training courses, and following the general procedures of the program. However, the level of implementation varied considerably between the municipalities, particularly with respect to fidelity (defined as implementation consistent with the principles of the interdisciplinary RTW process). Five municipalities had high and eight had low fidelity scores. Similar large differences were found with regard to dose-delivered, particularly in the quality of cooperation with beneficiaries, employers, and general practitioners. Only 50% of the first consultations with the RTW coordinator were conducted in time. Among participants who were employed when their sickness absence period started, only 9% had at least one meeting with their workplace. It was feasible to implement the basic features of the Danish RTW program, however, large variations existed between municipalities. Establishment of well-functioning interdisciplinary RTW teams might require more time and resources, while ensuring early assessment and more frequent cooperation with employers might need more general adjustments in the Danish sickness benefit system.


Successful periodontal ligament regeneration by periodontal progenitor preseeding on natural tooth root surfaces.
The regeneration of lost periodontal ligament (PDL) and alveolar bone is the purpose of periodontal tissue engineering. The goal of the present study was to assess the suitability of 3 odontogenic progenitor populations from dental pulp, PDL, and dental follicle for periodontal regeneration when exposed to natural and synthetic apatite surface topographies. We demonstrated that PDL progenitors featured higher levels of periostin and scleraxis expression, increased adipogenic and osteogenic differentiation potential, and pronounced elongated cell shapes on barren root chips when compared with dental pulp and dental follicle cells. When evaluating the effect of surface characteristics on PDL progenitors, natural root surfaces resulted in elongated PDL cell shapes, whereas PDL progenitors on synthetic apatite surfaces were rounded or polygonal. In addition, surface coatings affected PDL progenitor gene expression profiles: collagen I coatings enhanced alkaline phosphatase and osteocalcin expression levels and laminin-1 coatings increased epidermal growth factor (EGF), nestin, cadherin 1, and keratin 8 expression. PDL progenitors seeded on natural tooth root surfaces in organ culture formed new periodontal fibers after 3 weeks of culture. Finally, replantation of PDL progenitor-seeded tooth roots into rat alveolar bone sockets resulted in the complete formation of a new PDL and stable reattachment of teeth over a 6-month period. Together, these findings indicate that periodontal progenitor cell type as well as mineral surface topography and molecular environment play crucial roles in the regeneration of true periodontal anchorage.


An essential heparin-binding domain in the fibroblast growth factor receptor kinase.
Heparin or heparin-like heparan sulfate proteoglycans are obligatory for activity of the heparin-binding fibroblast growth factor (FGF) family. Heparin interacts independently of FGF ligand with a specific sequence (K18K) in one of the immunoglobulin-like loops in the extracellular domain of the FGF receptor tyrosine kinase transmembrane glycoprotein. A synthetic peptide corresponding to K18K inhibited heparin and heparin-dependent FGF binding to the receptor. K18K and an antibody to K18K were antagonists of FGF-stimulated cell growth. Point mutations of lysine residues in the K18K sequence abrogated both heparin- and ligand-binding activities of the receptor kinase. The results indicate that the FGF receptor is a ternary complex of heparan sulfate proteoglycan, tyrosine kinase transmembrane glycoprotein, and ligand.


Preliminary analysis of the effect of Stevia (Stevia rebaudiana) in patients with chronic kidney disease (stage I to stage III).
Stevia, Stevia rebaudiana (Bertoni), has become an important economic plant for its commercial use as a sweetener. Stevia plays a significant role in the healthcare practice of different cultures and in population. Previous animal and clinical studies demonstrated the efficacy of Stevia against chronic diseases like diabetes and hypertension. This study aimed to investigate the beneficial effect of Stevia in chronic kidney disease (CKD) patients after three (3) months of treatment along with the conventional antihypertensive and anti diabetic medications. A prospective, interventional, randomized, single-blind, placebo-controlled trial has been done with 97 participants. Stevia capsule (250 mg) or matching placebo was given to the participants twice daily along with Angiotensin-II Receptor Blocker (ARB) and/or Ca2+ Channel Blocker (CCB). First follow up visits were done after 3 months of the interval. Blood and urine samples were collected for the biochemical tests. A structured questionnaire was used for the baseline assessment. Informed consent was taken from each participant. Both hypertension and diabetes were found to be associated with CKD. Most of the participants (52.3%) of Stevia group were in CKD Stage II. Significant changes were found in Serum creatinine (p < 0.027), Serum Uric acid (p < 0.009), Fasting blood sugar (p < 0.041) and Postprandial blood sugar (p < 0.013) and Microalbumin (p < 0.041) level in the treatment group. The initial result demonstrated that Stevia has the potential for a significant improvement of some biochemical parameters in CKD patients. After completion of the nine (9) months clinical trial, the constructive effect of Stevia can be confirmed in this group of patients.


Thermal memory in self-assembled collagen fibril networks.
Collagen fibrils form extracellular networks that regulate cell functions and provide mechanical strength to tissues. Collagen fibrillogenesis is an entropy-driven process promoted by warming and reversed by cooling. Here, we investigate the influence of noncovalent interactions mediated by the collagen triple helix on fibril stability. We measure the kinetics of cold-induced disassembly of fibrils formed from purified collagen I using turbimetry, probe the fibril morphology by atomic force microscopy, and measure the network connectivity by confocal microscopy and rheometry. We demonstrate that collagen fibrils disassemble by subunit release from their sides as well as their ends, with complex kinetics involving an initial fast release followed by a slow release. Surprisingly, the fibrils are gradually stabilized over time, leading to thermal memory. This dynamic stabilization may reflect structural plasticity of the collagen fibrils arising from their complex structure. In addition, we propose that the polymeric nature of collagen monomers may lead to slow kinetics of subunit desorption from the fibril surface. Dynamic stabilization of fibrils may be relevant in the initial stages of collagen assembly during embryogenesis, fibrosis, and wound healing. Moreover, our results are relevant for tissue repair and drug delivery applications, where it is crucial to control fibril stability.


Association of interleukin-1 beta (-511C/T) polymorphisms with osteoporosis in postmenopausal women.
Osteoporosis is a common disease of the elderly, in which genetic and clinical factors contribute to the disease phenotype. Since the production of interleukin-1 (IL-1) has been implicated in the bone mass and skeletal disorders, we investigated whether IL-1 system gene polymorphisms are associated with the pathogenesis of osteoporosis in postmenopausal Taiwanese women. Osteoporosis is diagnosed by dual-energy x-ray absorptiometry, which measures bone mineral density (BMD) at multiple skeletal sites. We studied the IL-1α (-889C/T), IL-1β (-511C/T) and the 86 base pair variable number tandem repeat (VNTR) in intron 2 of the IL-1 receptor antagonist (IL-1ra) gene in 117 postmenopausal women with osteoporosis and 135 control subjects without a history of symptomatic osteoporosis. These gene polymorphisms were analyzed by polymerase chain reaction and restriction fragment length polymerase. Blood sugar and other risk factors were also determined. The frequencies of IL-1β (-511C/T) genotypes (P=.022, odds ratio=1.972) and alleles (P=.02, odds ratio=2.909) showed a statistically significant difference between the two groups. However, we did not find any statistically significant difference in IL-1β and IL-1ra polymorphisms (P>.05). We also observed a positive relationship between osteoporosis and cholesterol and a weak inverse relationship between blood sugar and osteoporosis in postmenopausal women. These experimental results suggest that the pathogenesis of osteoporosis is associated with IL-1β (-511C/T) polymorphism in postmenopausal women. This polymorphism is an independent risk factor for osteoporosis.


Organic solvent neurotoxicity. Facts and research needs.
While many organic solvents are, in large doses, capable of inducing an acute, reversible narcotic state, few unequivocally induce chronic, long-lasting, or irreversible changes in nervous system structure and/or function. For organic solvents with proved neurotoxic properties, the type of neurological damage is closely related to the structure of the chemical agent, while the degree of impairment and the extent of reversibility are related to the potency, dose, and duration of exposure. Examples include solvents containing n-hexane or methyl n-butyl ketone, which have caused many cases of occupational neuropathy. Chronic inhalation abuse of pure toluene produces irreversible cerebellar, brainstem, and pyramidal-tract dysfunction, but comparable changes have not been found in solvent workers occupationally exposed to toluene. Ototoxicity is found in experimental animals exposed to toluene, xylene, or styrene. Impure trichloroethylene has a predilection for damaging the trigeminal nerve; dichloroacetylene, a breakdown product of trichloroethylene, is probably responsible for this neurotoxic property. Prolonged occupational exposure to mixed solvents, notably white spirit, has been reported to induce a mild, nonprogressive dementing illness with or without peripheral nerve dysfunction, but supporting data from neuropathological and experimental animal studies are lacking. Many other solvents have been reported to induce adverse effects in workers. The pivotal biological role of the nervous system and its vulnerability to selected organic solvents widely used in industry underline the urgent need for further clinical and experimental research on this problem.


Influence of excitatory amino acids and ischemia on rat retinal choline acetyltransferase-containing cells.
To compare the effects of glutamate agonists and different types of ischemic insult on choline acetyltransferase (ChAT) immunoreactivity in the rat retina. Rat retinas were exposed to different glutamate agonists in vivo or in vitro for specific periods of time, and the retinas were then fixed and processed for the localization of ChAT immunoreactivity. In other experiments, rats were administered an ischemic insult either by ligaturing the carotids (two-vessel occlusion [2-VO] procedure), cannulating the anterior chamber, and raising the intraocular pressure (high intraocular pressure [HIOP] procedure) or placing a ligature around the optic nerve sufficiently tightly to prevent blood flow through the central retinal artery (vascular ligation [VL] procedure). The electroretinogram was recorded, and, after a specific period of time, reperfusion was allowed to occur. Thirty to 36 hours after reperfusion, the retinas were dissected and processed for the localization of ChAT, as well as for parvalbumin, Thy-1, and alpha PKC immunoreactivities. Of the glutamate agonists tested, only kainate reduced ChAT immunoreactivity significantly in vivo and in vitro. This effect of kainate could be counteracted by the antagonist CNQX (6-cyano-2,3-dihydroxy-7-nitroquinoxaline-2,3-dione). The ChAT immunoreactivity was unaffected in retinas in which ischemia was induced by the 2-VO procedure. In contrast, ChAT immunoreactivity was obliterated in retinas in which the HIOP was used and drastically reduced when the VL procedure was used. Interestingly, neither alpha PKC nor Thy-1 immunoreactivities were affected in retinas subjected to HIOP or VL methods. However, parvalbumin immunoreactivity was reduced in the HIOP model but only slightly altered in the VL model. The current results suggest that kainate receptors are associated with the cholinergic retinal neurones in the rat retina. Activation of these receptors by kainate causes a reduction in the neurones' ChAT content. This effect can be mimicked by subjecting the retina to a sufficiently harsh ischemic insult, as occurs in the VL and HIOP procedures. When the ischemic insult is mild, as in the 2-VO procedure, no obvious change in ChAT immunoreactivity is apparent. The HIOP procedure for inducing an ischemic insult was found to be the most severe of the three procedures used, because ChAT immunoreactivity was obliterated and clear changes in the parvalbumin immunoreactivity also were recorded. Interestingly, neither the HIOP nor the VL procedures caused a change in the Thy-1 and alpha PKC immunoreactivities.


TB research at UT-Houston--a review of cord factor: new approaches to drugs, vaccines and the pathogenesis of tuberculosis.
Tuberculosis remains a major threat as drug resistance continues to increase. Pulmonary tuberculosis in adults is responsible for 80% of clinical cases and nearly 100% of transmission of infection. Unfortunately, since we have no animal models of adult type pulmonary tuberculosis, the most important type of disease remains largely out of reach of modern science and many fundamental questions remain unanswered. This paper reviews research dating back to the 1950's providing compelling evidence that cord factor (trehalose 6,6 dimycolate [TDM]) is essential for understanding tuberculosis. However, the original papers by Bloch and Noll were too far ahead of their time to have immediate impact. We can now recognize that the physical and biologic properties of cord factor are unprecedented in science, especially its ability to switch between two sets of biologic activities with changes in conformation. While TDM remains on organisms, it protects them from killing within macrophages, reduces antibiotic effectiveness and inhibits the stimulation of protective immune responses. If it comes off organisms and associates with lipid, TDM becomes a driver of tissue damage and necrosis. Studies emanating from cord factor research have produced (1) a rationale for improving vaccines, (2) an approach to new drugs that overcome natural resistance to antibiotics, (3) models of caseating granulomas that reproduce multiple manifestations of human tuberculosis. (4) evidence that TDM is a key T cell antigen in destructive lesions of tuberculosis, and (5) a new understanding of the pathology and pathogenesis of postprimary tuberculosis that can guide more informative studies of long standing mysteries of tuberculosis.


Theoretical Analysis of the Behavior of Mercury plus Thallium Iodide Arc Lamps with Variable Mercury Loadings.
A theoretical analysis of the behavior of 400-watt Hg + TlI arc lamps with variable mercury loadings is performed. It is quantitatively shown that, as the Hg loading is doubled, the following effects may be predicted: (a) the effective arc temperature decreases from 5100 K to 4800 K, (b) the effective radius increases by ~10%, (c) the escape factor for the imprisoned Tl 5350 A line increases ~40%, (d) the power radiated in the Tl 5350 A line increases ~10%, and (e) the power radiated in the Hg visible line decreases ~15%. These results are in agreement with the experimental observations of Larson. These effects result in an increase in the lumen-per-watt ratings, which is of technological significance. Self-absorption is an important factor in the radiation of the Tl 5350 A line, and it is shown that the experimental observations are reasonably consistent with the assumption of local thermodynamic equilibrium.


The biological activities of (1,3)-(1,6)-beta-d-glucan and porous electrospun PLGA membranes containing beta-glucan in human dermal fibroblasts and adipose tissue-derived stem cells.
In this study, we investigated the possible roles of (1,3)-(1,6)-beta-d-glucan (beta-glucan) and porous electrospun poly-lactide-co-glycolide (PLGA) membranes containing beta-glucan for skin wound healing, especially their effect on adult human dermal fibroblast (aHDF) and adipose tissue-derived stem cell (ADSC) activation, proliferation, migration, collagen gel contraction and biological safety tests of the prepared membrane. This study demonstrated that beta-glucan and porous PLGA membranes containing beta-glucan have enhanced the cellular responses, proliferation and migration, of aHDFs and ADSCs and the result of a collagen gel contraction assay also revealed that collagen gels contract strongly after 4 h post-gelation incubation with beta-glucan. Furthermore, we confirmed that porous PLGA membranes containing beta-glucan are biologically safe for wound healing study. These results indicate that the porous PLGA membranes containing beta-glucan interacted favorably with the membrane and the topical administration of beta-glucan was useful in promoting wound healing. Therefore, our study suggests that beta-glucan and porous PLGA membranes containing beta-glucan may be useful as a material for enhancing wound healing.


Changes in expression and honesty of sexual signalling over the reproductive lifetime of sticklebacks.
Fitness costs of signalling are essential in order for reliable sexual signalling to prevail when the interests of the sexes conflict. This means that signalling can be subjected to a life history trade-off between present and future signalling effort. Here, I show that three-spined stickleback males (Gasterosteus aculeatus), who have a single breeding season during which they breed repeatedly, change their red nuptial coloration over the season depending on their body size at the start of breeding. Large males that completed several breeding cycles increased their red coloration over the season, whereas small males, who completed only a few cycles, did not. The increase in coloration was accompanied by an increase in parental success when males were energy constrained, but not when they had access to an unlimited food supply. Red coloration was thus an honest signal of male parental ability despite changes in signal expression when both signalling and parental care were costly and the investments in them changed simultaneously over the reproductive lifetime. However, the honesty of the signal varied over a lifetime. At the penultimate cycle, bright males cannibalized some of their eggs, probably to increase survival to the last cycle, whereas males cared for their offspring independent of coloration at the ultimate cycle.


Concentrations and emission rates of aerial ammonia, nitrous oxide, methane, carbon dioxide, dust and endotoxin in UK broiler and layer houses.
1. A survey of the concentration and emission rates of aerial ammonia, nitrous oxide, methane, carbon dioxide, dust and endotoxin was undertaken in 4 examples each of typical UK broiler, cage and perchery houses over 24 h during winter and summer. 2. Overall the air quality within the poultry houses was unsatisfactory as judged by the dual criteria of farmer health and bird performance. 3. Mean concentrations of ammonia ranged from 12.3 to 24.2 ppm while concentrations of methane and nitrous oxide were close to ambient levels. Mass concentrations of aerial dust ranged from 2 to 10 mg/m3 and 0.3 to 1.2 mg/m3 for inspirable and respirable fractions respectively, while endotoxin concentration was typically about 0.1 microgram/m3 (inspirable fraction). 4. Emission rates of gaseous ammonia were rapid (9.2 g (NH3)/h per 500 kg live body weight) and uniform across the three types of building, while emissions of methane and nitrous oxide were slow. Rates of dust emission ranged from 0.86 to 8.24 g/h per 500 kg live body weight in the inspirable size fraction.


Subunit-specific gating controls rat NR1/NR2A and NR1/NR2B NMDA channel kinetics and synaptic signalling profiles.
NR2A and NR2B are the predominant NR2 NMDA receptor subunits expressed in cortex and hippocampus. The relative expression level of NR2A and NR2B is regulated developmentally and these two subunits have been suggested to play distinct roles in long-term synaptic plasticity. We have used patch-clamp recording of recombinant NMDA receptors expressed in HEK293 cells to characterize the activation properties of both NR1/NR2A and NR1/NR2B receptors. Recordings from outside-out patches that contain a single active channel show that NR2A-containing receptors have a higher probability of opening at least once in response to a brief synaptic-like pulse of glutamate than NR2B-containing receptors (NR2A, 0.80; NR2B, 0.56), a higher peak open probability (NR2A, 0.50; NR2B, 0.12), and a higher open probability within an activation (NR2A, 0.67; NR2B, 0.37). Analysis of the sequence of single-channel open and closed intervals shows that both NR2A- and NR2B-containing receptors undergo multiple conformational changes prior to opening of the channel, with at least one of these steps being faster for NR2A than NR2B. These distinct properties produce profoundly different temporal signalling profiles for NR2A- and NR2B-containing receptors. Simulations of synaptic responses demonstrate that at low frequencies typically used to induce long-term depression (LTD; 1 Hz), NR1/NR2B makes a larger contribution to total charge transfer and therefore calcium influx than NR1/NR2A. However, under high-frequency tetanic stimulation (100 Hz; > 100 ms) typically used to induce long-term potentiation (LTP), the charge transfer mediated by NR1/NR2A considerably exceeds that of NR1/NR2B.


The Perruche judgment and the "right not to be born".
The French government has given in to public pressure and overturned a controversial legal ruling which recognised the right of a disabled child to seek damages. Most notably, the ruling, widely described as establishing a child's right "not to be born", had provoked "outrage" amongst groups defending the rights of the disabled and led to a ban on prenatal scans by French gynaecologists. Once again, only parents will be able to seek damages but some people think the ruling has been misinterpreted.


Challenges in assessing release, exposure and fate of silver nanoparticles within the UK environment.
There are significant challenges in assessing the fate and exposure of nanoparticles (NPs) in the environment owing to the lack of information on their use, potential pathways and sinks in the environment. In order to better understand the environmental exposure of silver nanoparticles (AgNPs), we reviewed the uses and potential exposure sources of both Ag and AgNPs. The approach taken was to identify the range of products that utilise its properties, identify potential environmental release pathways and subsequent fate once released into the environment. We then compared measured environmental concentrations with modelled concentrations from our work and others. We estimate that within the United Kingdom (UK) a total quantity of 8.8 tonnes per year of AgNPs is released from products containing AgNPs and enters UK waste water systems. Sewage sludge was identified as an important receiving compartment. Agricultural land with sludge applied was estimated to have a yearly increase in AgNP concentration of 36 μg per kg per year. Available ecotoxicity data for soil and the predicted environmental concentrations are used to perform a risk characterisation. This work highlights the on-going challenges faced when undertaking a risk assessment of AgNPs in the environment.


HIV Among MSM and Heterosexual Women in the United States: An Ecologic Analysis.
Phylogenetic studies show links between heterosexual women and men who have sex with men (MSM) that are more numerous than from heterosexual men to women suggesting that HIV infections among heterosexual women may stem from MSM. Poor communities have been associated with high rates of HIV among heterosexual women. Our analysis investigates potential transmission of HIV between MSM and female heterosexuals. National HIV Behavioral Surveillance data describe transmission risk behaviors of MSM, and HIV case reporting data describe the percentages of cases that are attributed to transmission risk categories. We examined correlations between the percentages of men who were MSM who also have sex with women and female heterosexual cases. We also examined census data to characterize each city in terms of poverty level and race/ethnicity makeup. There was a high correlation (0.93) between the percentage of reported living HIV cases attributed to male heterosexual contact and female heterosexual contact and a moderate nonsignificant correlation (0.49) between the percentage of MSM who were men who have sex with men and women (MSMW) in National HIV Behavioral Surveillance and the percentage of reported cases that were attributed to female heterosexual contact suggesting some potential overlap. Cities with high levels of poverty and African American/Black residents had higher levels of MSMW and higher levels of heterosexual female cases. Addressing HIV in cities with high levels of MSMW may have the dual effect of improving the health of MSM populations that have a high burden of HIV and to improve the health of their larger communities.


Irradiation-induced switch power drift in optically bistable nonlinear interference filters at 514 and 830 nm.
Light-induced changes in the switch power of optically bistable ZnSe nonlinear interference filters have been studied experimentally at 514 and 830 nm wavelengths. This drift was found to be mainly caused by an irreversible change in the peak wavelength and transmission of the filter and is associated with high internal operating irradiances. These changes are apparently due to photostructural modifications of the material forming the central spacer layer. The drift was shown to be dependent on the spacer thickness, incident spot size and the deposition technique used to grow the filter structures. The rate of switch power drift was also found to be reduced considerably when using an illuminating wavelength well removed from the band edge of the spacer material, e.g., 830 nm.


Perceived object stability depends on multisensory estimates of gravity.
How does the brain estimate object stability? Objects fall over when the gravity-projected centre-of-mass lies outside the point or area of support. To estimate an object's stability visually, the brain must integrate information across the shape and compare its orientation to gravity. When observers lie on their sides, gravity is perceived as tilted toward body orientation, consistent with a representation of gravity derived from multisensory information. We exploited this to test whether vestibular and kinesthetic information affect this visual task or whether the brain estimates object stability solely from visual information. In three body orientations, participants viewed images of objects close to a table edge. We measured the critical angle at which each object appeared equally likely to fall over or right itself. Perceived gravity was measured using the subjective visual vertical. The results show that the perceived critical angle was significantly biased in the same direction as the subjective visual vertical (i.e., towards the multisensory estimate of gravity). Our results rule out a general explanation that the brain depends solely on visual heuristics and assumptions about object stability. Instead, they suggest that multisensory estimates of gravity govern the perceived stability of objects, resulting in objects appearing more stable than they are when the head is tilted in the same direction in which they fall.


CD4-induced activation in a soluble HIV-1 Env trimer.
The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion.


Uncovering global SUMOylation signaling networks in a site-specific manner.
SUMOylation is a reversible post-translational modification essential for genome stability. Using high-resolution MS, we have studied global SUMOylation in human cells in a site-specific manner, identifying a total of >4,300 SUMOylation sites in >1,600 proteins. To our knowledge, this is the first time that >1,000 SUMOylation sites have been identified under standard growth conditions. We quantitatively studied SUMOylation dynamics in response to SUMO protease inhibition, proteasome inhibition and heat shock. Many SUMOylated lysines have previously been reported to be ubiquitinated, acetylated or methylated, thus indicating cross-talk between SUMO and other post-translational modifications. We identified 70 phosphorylation and four acetylation events in proximity to SUMOylation sites, and we provide evidence for acetylation-dependent SUMOylation of endogenous histone H3. SUMOylation regulates target proteins involved in all nuclear processes including transcription, DNA repair, chromatin remodeling, precursor-mRNA splicing and ribosome assembly.


Flavanoids induce expression of the suppressor of cytokine signalling 3 (SOCS3) gene and suppress IL-6-activated signal transducer and activator of transcription 3 (STAT3) activation in vascular endothelial cells.
The atherogenic cytokine IL-6 (interleukin-6) induces pro-inflammatory gene expression in VECs (vascular endothelial cells) by activating the JAK (Janus kinase)/STAT3 (signal transducer and activator of transcription 3) signalling pathway, which is normally down-regulated by the STAT3-dependent induction of the E3 ubiquitin ligase component SOCS3 (suppressor of cytokine signalling 3). Novel treatments based on the regulation of SOCS3 protein levels could therefore have value in the treatment of diseases with an inflammatory component, such as atherosclerosis. To this end we carried out a screen of 1031 existing medicinal compounds to identify inducers of SOCS3 gene expression and identified the flavanoids naringenin and flavone as effective inducers of SOCS3 protein, mRNA and promoter activity. This was in contrast with the action of traditional JAK/STAT3 inhibitors and the polyphenol resveratrol, which effectively suppress SOCS3 gene expression. Both naringenin and flavone also effectively suppressed IL-6-stimulated phosphorylation of STAT3 (Tyr⁷⁰⁵) which led to suppression of IL-6-induction of the atherogenic STAT3 target gene MCP1 (monocyte chemotactic protein-1), suggesting that their ability to induce SOCS3 gene expression is STAT3-independent. Supporting this idea was the observation that the general kinase inhibitor compound C inhibits flavone- and cAMP-dependent, but not JAK-dependent, SOCS3 induction in VECs. Indeed, the ability of flavanoids to induce SOCS3 expression requires activation of the ERK (extracellular-signal-regulated kinase)-dependent transcription factor SP3, and not STAT3. In the present paper we therefore describe novel molecular actions of flavanoids, which control SOCS3 gene induction and suppression of STAT3 signalling in VECs. These mechanisms could potentially be exploited to develop novel anti-atherogenic therapies.


The failure of preoperative ethylene-vinyl alcohol copolymer embolization to improve outcomes in arteriovenous malformation management: case series.
Ethylene-vinyl alcohol copolymer embolization is increasingly used preoperatively in the resection of brain arteriovenous malformations (AVMs). However, the case for embolization improving the outcome of resection has not been evaluated. In this paper the authors set out to compare outcomes after surgery for brain AVMs in 2 consecutive periods of practice. In the first period, selective embolization was used without the use of ethylene-vinyl alcohol copolymer. In the second period, selective embolization with ethylene-vinyl alcohol copolymer was performed. A consecutive case series (prospectively collected data) was retrospectively analyzed. Adverse outcomes were considered to be an outcome modified Rankin Scale score greater than 2 due to embolization or surgery. A total of 538 surgical cases were included. The percentages of adverse outcomes were as follows: 0.34% for Spetzler-Martin AVMs less than Grade III (1 of 297 cases); 5.23% (95% CI 2.64%-9.78%) for Grade III AVMs (9 of 172 cases); and 17% (95% CI 10%-28%) for AVMs greater than Grade III (12 of 69 cases). There was no improvement in outcomes from the first period to the second period. The adverse outcome for Grade III brain AVMs in the first period was 5.2% (7 of 135 cases) and in the second period (after ethylene-vinyl alcohol copolymer was introduced) it was 5.4% (2 of 37 cases). For AVMs greater than Grade III, the adverse outcome was 12% (6 of 49 cases) in the first period and 30% (6 of 20 cases) in the second period. Outcomes for brain AVM surgery were not improved by ethylene-vinyl alcohol copolymer embolization. Preoperative embolization of high-grade AVMs with an ethylene-vinyl alcohol copolymer did not prevent those hemorrhagic complications which embolization is hypothesized to prevent based on theoretical speculations but not demonstrated in practice.


Efficacy of maternal B12 supplementation in vegetarian women for improving infant neurodevelopment: protocol for the MATCOBIND multicentre, double-blind, randomised controlled trial.
Vitamin B12 deficiency is widely prevalent across many low- and middle-income countries, especially where the diet is low in animal sources. While many observational studies show associations between B12 deficiency in pregnancy and infant cognitive function (including memory, language and motor skills), evidence from clinical trials is sparse and inconclusive. This double-blind, multicentre, randomised controlled trial will enrol 720 vegetarian pregnant women in their first trimester from antenatal clinics at two hospitals (one in India and one in Nepal). Eligible mothers who give written consent will be randomised to receive either 250 mcg methylcobalamin or 50 mcg (quasi control), from enrolment to 6 months post-partum, given as an oral daily capsule. All mothers and their infants will continue to receive standard clinical care. The primary trial outcome is the offspring's neurodevelopment status at 9 months of age, assessed using the Development Assessment Scale of Indian Infants. Secondary outcomes include the infant's biochemical B12 status at age 9 months and maternal biochemical B12 status in the first and third trimesters. Maternal biochemical B12 status will also be assessed in the first trimester. Modification of association by a priori identified factors will also be explored. The study protocol has been approved by ethical committees at each study site (India and Nepal) and at University College London, UK. The study results will be disseminated to healthcare professionals and academics globally via conferences, presentations and publications. Researchers at each study site will share results with participants during their follow-up visits.Trial registration numberCTRI/2018/07/015048 (Clinical Trial Registry of India); NCT04083560 (ClinicalTrials.gov).


A tailored-guided internet-based cognitive-behavioral intervention for patients with rheumatoid arthritis as an adjunct to standard rheumatological care: results of a randomized controlled trial.
For patients with chronic pain conditions such as rheumatoid arthritis (RA), who experience elevated levels of distress, tailored-guided internet-based cognitive-behavioral treatment may be effective in improving psychological and physical functioning, and reducing the impact of RA on daily life. A multicenter, randomized controlled trial was conducted for RA patients with elevated levels of distress as assessed by a disease-specific measure. The control group (n = 71) received standard care and the intervention group (n = 62) additionally received an internet-based tailored cognitive-behavioral intervention. Main analyses were performed using a linear mixed model estimating differences between the intervention and control groups in scores of psychological functioning, physical functioning, and impact of RA on daily life at preassesment and postassessment, and at 3, 6, 9, and 12 months. Patients who received the internet-based intervention reported a larger improvement in psychological functioning compared with the control group, indicating less depressed mood (P < 0.001, d = 0.54), negative mood (P = 0.01, d = 0.38), and anxiety (P < 0.001, d = 0.48) during the course of the 1-year follow-up period. Regarding physical functioning, a trend was found for the intervention group reporting less fatigue than the control group (P = 0.06, d = 0.24), whereas no effect was found on pain. No effects were found for the impact of RA on daily life, except for the intervention group experiencing fewer role limitations due to emotional problems (P < 0.001, d = 0.53). Offering guided internet-based cognitive-behavioral therapy is a promising development to aid patients with psychological distress particularly in improving psychological functioning. Further research on adherence and specific intervention ingredients is warranted.


A classification of vulvoscopic findings for clinical diagnosis.
We describe vulvoscopic findings, their correspondence with certain pathological processes, and the grouping of vulvoscopic findings to facilitate clinical practice. We performed vulvoscopy on 2,352 patients and described what we saw. We then classified the images by groups, according to the diagnoses suggested. Finally, we compared the frequency of the different images as a function of the symptoms, the clinical and histological diagnoses, the history of cervical lesions (cervical human papillomavirus, cervical intraepithelial neoplasia, or cervical carcinoma), and the microbiological studies. We found 2.33% human papillomavirus-vulvar intraepithelial neoplasia (HPV-VIN) type 1 lesions, 0.52% herpes simplex lesions, and 0.08% VIN2 lesions in asymptomatic patients without a history of cervical lesions. In 95% of patients in whom raised acetowhite patches were found on the skin, vaginal exudate revealed Candida species. The images showing cobbled mucosa and white and red punctation primarily suggested a nonviral infection, though a small percentage of cases were of viral origin. The history of cervical lesions was significant (p < .000001) in the HPV-VIN1 lesion group. For the diagnosis of more severe vulvar lesions (VIN3 and vulvar carcinoma), we found that the joint presence of pruritus and a history of cervical lesions was significant (p < .004). Vulvoscopy performed as a routine examination procedure enables the detection of some disorders in asymptomatic patients who lack a history of cervical lesions and precludes the need for histological studies in some cases, though in others the image is inconclusive and a biopsy is required. Given that a description of normal findings is available, we are able to eliminate suspect pathological processes through a vulvoscopic examination. The proposed classification can serve as a guide to diagnosis and therapy.


A new mouse model for evaluating the immunotherapy of human colorectal cancer.
A new murine model of human colorectal cancer was generated by crossing human carcinoembryonic antigen (CEA) transgenic mice (H-2K(b)) with adenomatous polyposis coli (Apc1638N) knockout mice (H-2K(b)). The resulting hybrid mice developed gastrointestinal polyps in 6-8 months that progressed to invasive carcinomas with a similar pattern of dysplasia and CEA expression as observed in human colorectal cancer. These animals exhibited incomplete or partial tolerance to CEA as evidenced by delayed growth of CEA-expressing tumors and the inability to inhibit CEA-specific CTL responses. These results have important implications for understanding the role of CEA-specific immunity in human colon cancer patients and suggest that vaccine strategies targeting CEA may be feasible. This model provides a powerful system for evaluating antigen-specific tumor immunity against spontaneous tumors arising in an orthotopic location and permits evaluation of therapeutic vaccine strategies for human colorectal cancer.


Focused Transhepatic Electroporation Mediated by Hypersaline Infusion through the Portal Vein in Rat Model. Preliminary Results on Differential Conductivity.
Spread hepatic tumours are not suitable for treatment either by surgery or conventional ablation methods. The aim of this study was to evaluate feasibility and safety of selectively increasing the healthy hepatic conductivity by the hypersaline infusion (HI) through the portal vein. We hypothesize this will allow simultaneous safe treatment of all nodules by irreversible electroporation (IRE) when applied in a transhepatic fashion. Sprague Dawley (Group A, n = 10) and Athymic rats with implanted hepatic tumour (Group B, n = 8) were employed. HI was performed (NaCl 20%, 3.8 mL/Kg) by trans-splenic puncture. Deionized serum (40 mL/Kg) and furosemide (2 mL/Kg) were simultaneously infused through the jugular vein to compensate hypernatremia. Changes in conductivity were monitored in the hepatic and tumour tissue. The period in which hepatic conductivity was higher than tumour conductivity was defined as the therapeutic window (TW). Animals were monitored during 1-month follow-up. The animals were sacrificed and selective samples were used for histological analysis. The overall survival rate was 82.4% after the HI protocol. The mean maximum hepatic conductivity after HI was 2.7 and 3.5 times higher than the baseline value, in group A and B, respectively. The mean maximum hepatic conductivity after HI was 1.4 times higher than tumour tissue in group B creating a TW to implement selective IRE. HI through the portal vein is safe when the hypersaline overload is compensated with deionized serum and it may provide a TW for focused IRE treatment on tumour nodules.


Accounting for respiratory motion in small serial structures during radiotherapy planning: proof of concept in virtual bronchoscopy-guided lung functional avoidance radiotherapy.
Respiratory motion management techniques in radiotherapy (RT) planning are primarily focused on maintaining tumor target coverage. An inadequately addressed need is accounting for motion in dosimetric estimations in smaller serial structures. Accurate dose estimations in such structures are more sensitive to motion because respiration can cause them to move completely in or out of a high dose-gradient field. In this work, we study three motion management strategies (m1-m3) to find an accurate method to estimate the dosimetry in airways. To validate these methods, we generated a 'ground truth' digital breathing model based on a 4DCT scan from a lung stereotactic ablative radiotherapy (SAbR) patient. We simulated 225 breathing cycles with ±10% perturbations in amplitude, respiratory period, and time per respiratory phase. A high-resolution breath-hold CT (BHCT) was also acquired and used with a research virtual bronchoscopy software to autosegment 239 airways. Contours for planning target volume (PTV) and organs at risk (OARs) were defined on the maximum intensity projection of the 4DCT (CTMIP) and transferred to the average of the 10 4DCT phases (CTAVG). To design the motion management methods, the RT plan was recreated using different images and structure definitions. Methods m1 and m2 recreated the plan using the CTAVG image. In method m1, airways were deformed to the CTAVG. In m2, airways were deformed to each of the 4DCT phases, and union structures were transferred onto the CTAVG. In m3, the RT plan was recreated on each of the 10 phases, and the dose distribution from each phase was deformed to the BHCT and summed. Dose errors (mean [min, max]) in airways were: m1: 21% (0.001%, 93%); m2: 45% (0.1%, 179%); and m3: 4% (0.006%, 14%). Our work suggests that accurate dose estimation in moving small serial structures requires customized motion management techniques (like m3 in this work) rather than current clinical and investigational approaches.


Detection of vancomycin-resistant Enterococcus faecium hospital-adapted lineages in municipal wastewater treatment plants indicates widespread distribution and release into the environment.
Vancomycin-resistant Enterococcus faecium (VREfm) is a leading cause of healthcare-associated infection. Reservoirs of VREfm are largely assumed to be nosocomial although there is a paucity of data on alternative sources. Here, we describe an integrated epidemiological and genomic analysis of E. faecium associated with bloodstream infection and isolated from wastewater. Treated and untreated wastewater from 20 municipal treatment plants in the East of England, United Kingdom was obtained and cultured to isolate E. faecium, ampicillin-resistant E. faecium (AREfm), and VREfm. VREfm was isolated from all 20 treatment plants and was released into the environment by 17/20 plants, the exceptions using terminal ultraviolet light disinfection. Median log10 counts of AREfm and VREfm in untreated wastewater from 10 plants in direct receipt of hospital sewage were significantly higher than 10 plants that were not. We sequenced and compared the genomes of 423 isolates from wastewater with 187 isolates associated with bloodstream infection at five hospitals in the East of England. Among 481 E. faecium isolates belonging to the hospital-adapted clade, we observed genetic intermixing between wastewater and bloodstream infection, with highly related isolates shared between a major teaching hospital in the East of England and 9/20 plants. We detected 28 antibiotic resistance genes in the hospital-adapted clade, of which 23 were represented in bloodstream, hospital sewage, and municipal wastewater isolates. We conclude that our findings are consistent with widespread distribution of hospital-adapted VREfm beyond acute healthcare settings with extensive release of VREfm into the environment in the East of England.


The habu genome reveals accelerated evolution of venom protein genes.
Evolution of novel traits is a challenging subject in biological research. Several snake lineages developed elaborate venom systems to deliver complex protein mixtures for prey capture. To understand mechanisms involved in snake venom evolution, we decoded here the ~1.4-Gb genome of a habu, Protobothrops flavoviridis. We identified 60 snake venom protein genes (SV) and 224 non-venom paralogs (NV), belonging to 18 gene families. Molecular phylogeny reveals early divergence of SV and NV genes, suggesting that one of the four copies generated through two rounds of whole-genome duplication was modified for use as a toxin. Among them, both SV and NV genes in four major components were extensively duplicated after their diversification, but accelerated evolution is evident exclusively in the SV genes. Both venom-related SV and NV genes are significantly enriched in microchromosomes. The present study thus provides a genetic background for evolution of snake venom composition.


Performance of the FilmArray® blood culture identification panel utilized by non-expert staff compared with conventional microbial identification and antimicrobial resistance gene detection from positive blood cultures.
Utilization of commercially available rapid platforms for microbial identification from positive blood cultures is useful during periods of, or in laboratories with, limited expert staffing. We compared the results of the FilmArray® BCID Panel performed by non-expert technologists to those of conventional methods for organism identification performed by skilled microbiologists. Within 8 h of signalling positive by a continuous monitoring blood culture system, positive bottles were analysed by the FilmArray BCID Panel. Data from these analyses were compared to standard-of-care testing, which included conventional and automated methods. To gauge the ease of use of the BCID Panel by non-expert staff, technologists unfamiliar with diagnostic bacteriology performed the testing without prior knowledge of the Gram stain results, or even whether organisms were detected. Identifications of 172/200 (86 %) positive blood cultures using the BCID Panel were consistent with identifications provided by standard-of-care methods. Standard-of-care testing identified organisms in 20 positive blood cultures, which were not represented on the BCID Panel. Seven (3.5 %) blood cultures demonstrated a discrepancy between the methods, which could not be attributed to either a lack of representation on the panel or unclear separate detection of organisms in a mixed blood culture of a shared genus or grouping of organisms, e.g. Staphylococcus or Enterobacteriaceae . One (0.5 %) blood culture yielded invalid results on two separate panels, so it was eliminated from the study. The easy-to-use FilmArray® technology shows good correlation with blood culture identification and antibiotic resistance detection performed by conventional methods. This technology may be particularly useful in laboratories with limited staffing or limited technical expertise.


Gene circuit performance characterization and resource usage in a cell-free "breadboard".
The many successes of synthetic biology have come in a manner largely different from those in other engineering disciplines; in particular, without well-characterized and simplified prototyping environments to play a role analogous to wind-tunnels in aerodynamics and breadboards in electrical engineering. However, as the complexity of synthetic circuits increases, the benefits--in cost savings and design cycle time--of a more traditional engineering approach can be significant. We have recently developed an in vitro "breadboard" prototyping platform based on E. coli cell extract that allows biocircuits to operate in an environment considerably simpler than, but functionally similar to, in vivo. The simplicity of this system makes it a promising tool for rapid biocircuit design and testing, as well as for probing fundamental aspects of gene circuit operation normally masked by cellular complexity. In this work, we characterize the cell-free breadboard using real-time and simultaneous measurements of transcriptional and translational activities of a small set of reporter genes and a transcriptional activation cascade. We determine the effects of promoter strength, gene concentration, and nucleoside triphosphate concentration on biocircuit properties, and we isolate the specific contributions of essential biomolecular resources-core RNA polymerase and ribosomes-to overall performance. Importantly, we show how limits on resources, particularly those involved in translation, are manifested as reduced expression in the presence of orthogonal genes that serve as additional loads on the system.


Identification and characterization of genes encoding the human transferrin-binding proteins from Haemophilus influenzae.
Haemophilus influenzae, a strict human pathogen, acquires iron in vivo through the direct binding and removal of iron from human transferrin by an as yet uncharacterized process at the bacterial cell surface. In this study, the tbpA and tbpB genes of H. influenzae, encoding the transferrin-binding proteins Tbp1 and Tbp2, respectively, were cloned and sequenced. Alignments of the H. influenzae Tbp1 and Tbp2 protein sequences with those of related proteins from heterologous species were analyzed. On the basis of similarities between these and previously characterized proteins, Tbp1 appears to be a member of the TonB-dependent family of outer membrane proteins while Tbp2 is lipid modified by signal peptidase II. Isogenic mutants deficient in expression of Tbp1 or Tbp2 or both proteins were prepared by insertion of the Tn903 kanamycin resistance cassette into cloned sequences and reintroduction of the interrupted sequences into the wild-type chromosome. Binding assays with the mutants showed that a significant reduction in transferrin-binding ability resulted from the loss of either of the Tbps and a complete loss of binding was evident when neither protein was expressed. Loss of either Tbp2 or both proteins correlated with an inability to grow on media supplemented with transferrin-bound iron as the sole source of iron, whereas the Tbp1+ Tbp2- mutant was able to grow only at high transferrin concentrations.


Runx2 deletion in smooth muscle cells inhibits vascular osteochondrogenesis and calcification but not atherosclerotic lesion formation.
Vascular smooth muscle cells (SMCs) are major precursors contributing to osteochondrogenesis and calcification in atherosclerosis. Runt-related transcription factor-2 (Runx2) has been found essential for SMC differentiation to an osteochondrogenic phenotype and subsequent calcification in vitro. A recent study using a conditional targeting allele that produced a truncated Runx2 protein in SMCs of ApoE-/- mice showed reduced vascular calcification, likely occurring via reduction of receptor activator of nuclear factor-κB ligand (RANKL), macrophage infiltration, and atherosclerotic lesion formation. Using an improved conditional Runx2 knockout mouse model, we have elucidated new roles for SMC-specific Runx2 in arterial intimal calcification (AIC) without effects on atherosclerotic lesion size. We used an improved targeting construct to generate LDLr-/- mice with floxed-Runx2 alleles ( LDLr-/- :Runx2 f/f ) such that Cre-mediated recombination ( LDLr-/- :Runx2 ΔSM ) does not produce functional truncated Runx2 protein, thereby avoiding off-target effects. We found that both LDLr-/- :Runx2 f/f and LDLr-/- :Runx2 ΔSM mice fed with a high fat diet developed atherosclerosis. SMC-specific Runx2 deletion did not significantly reduce atherosclerotic lesion size, macrophage number, or expression of RANKL, MCP-1, and CCR2. However, it significantly reduced AIC by 50%. Mechanistically, Sox9 and type II collagen were unaltered in vessels of LDLr-/- :Runx2 ΔSM mice compared to LDLr-/- :Runx2 f/f counterparts, while type X collagen, MMP13 and the osteoblastic marker osteocalcin were significantly reduced. SMC autonomous Runx2 is required for SMC differentiation towards osteoblast-like cells, SMC-derived chondrocyte maturation and AIC in atherosclerotic mice. These effects were independent of systemic lipid metabolism, RANKL expression, macrophage infiltration, and atheromatous lesion progression.


Construction of a high resolution linkage disequilibrium map to evaluate common genetic variation in TP53 and neural tube defect risk in an Irish population.
Genetic and environmental factors contribute to the etiology of neural tube defects (NTDs). While periconceptional folic acid supplementation is known to significantly reduce the risk of NTDs, folate metabolic pathway related factors do not account for all NTDs. Evidence from mouse models indicates that the tumor protein p53 (TP53) is involved in implantation and normal neural tube development. To determine whether genetic variation in TP53 might contribute to NTD risk in humans, we constructed a high resolution linkage disequilibrium (LD) map of the TP53 genomic region based on genotyping 21 markers in an Irish population. We found that nine of these variants can be used to capture the majority of common variation in the TP53 genomic region. In contrast, the 3-marker haplotype commonly reported in the TP53 literature offers limited coverage of the variation in the gene. We used the expanded set of polymorphisms to measure the influence of TP53 on NTDs using both case-control and family based tests of association. We also assayed a functional variant in the p53 regulator MDM2 (rs2279744). Alleles of three noncoding TP53 markers were associated with NTD risk. A case effect was seen with the GG genotype of rs1625895 in intron 6 (OR = 1.37 [1.04-1.79], P = 0.02). A maternal effect was seen with the 135/135 genotype of the intron 1 VNTR (OR = 1.86 [1.16-2.96], P = 0.01) and the TT genotype of rs1614984 (RR = 0.58 [0.37-0.91], P = 0.02). As multiple comparisons were made, these cannot be considered definitive positive findings and additional investigation is required.


A comparison of entrance skin dose delivered by clinical angiographic c-arms using the real-time dosimeter: the MOSkin.
Coronary angiography is a procedure used in the diagnosis and intervention of coronary heart disease. The procedure is often considered one of the highest dose diagnostic procedures in clinical use. Despite this, there is minimal use of dosimeters within angiographic catheterisation laboratories due to challenges resulting from their implementation. The aim of this study was to compare entrance dose delivery across locally commissioned c-arms to assess the need for real-time dosimetry solutions during angiographic procedures. The secondary aim of this study was to establish a calibration method for the MOSkin dosimeter that accurately produces entrance dose values from the clinically sampled beam qualities and energies. The MOSkin is a real-time dosimeter used to measure the skin dose delivered by external radiation beams. The suitability of the MOSkin for measurements in the angiographic catheterisation laboratory was assessed. Measurements were performed using a 30 × 30 × 30 cm(3) PMMA phantom positioned at the rotational isocenter of the c-arm gantry. The MOSkin calibration factor was established through comparison of the MOSkin response to EBT2 film response. Irradiation of the dosimeters was performed using several clinical beam qualities ranging in energy from 70 to 105 kVp. A total of four different interventional c-arm machines were surveyed and compared using the MOSkin dosimeter. The phantom was irradiated from a normal angle of incidence using clinically relevant protocols, field sizes and source to image detector distance values. The MOSkin was observed to be radiotranslucent to the c-arm beam in all clinical environments. The MOSkin response was reproducible to within 2 % of the average value across repeated measurements for each beam setting. There were large variations in entrance dose delivery to the phantom between the different c-arm machines with the highest observed cine-acquisition entrance dose rate measuring 326 % higher than the lowest measured cine-acquisition entrance dose rate and with the highest measured fluoroscopic entrance dose rate measuring 346 % higher than the lowest measured fluoroscopic entrance dose rate. This comparison of entrance dose delivery across local clinical c-arms demonstrated the disparity in entrance dose delivery across catheterisation laboratories and outlined a need for real-time dose monitoring systems for patients during angiographic procedures. Through use of our calibration method, an average MOSkin calibration of 7.37 mV/cGy was established. The calibration method allowed entrance dose to be measured across a range of beam energies and beam qualities without the input of the c-arm beam characteristics. This calibration factor was proven to reproduce entrance dose values to within 5 % value of the reference dosimeter's response, suggesting potential for further studies and utilisation of the dosimeter in this field.


NM-23 gene loss of heterozygosity and protein expression in high-stage laryngeal squamous cell carcinomas.
Tumor suppressor genes that reduce metastatic potential have been described in a variety of different tumor types. One of the main tumor metastasis suppressor genes is nm-23, which is a nucleoside diphosphate kinase. Two isotypes, nm-23H1 and nm-23H2, have been cloned and map to chromosome 17q21.3. In a variety of tumors, including colon cancer and breast cancer, loss of expression of nm-23 is associated with lymph node metastasis. In other organ systems, however, this relationship is not seen. In head and neck squamous cell carcinomas (HNSCC), there have been conflicting results regarding the association between nm-23 protein expression and metastatic potential. To further explore the tumor metastasis suppressor function of nm-23 in HNSCC, we studied high-stage laryngeal carcinomas, tumors with and without cervical lymph node metastasis for nm-23 protein expression and loss of heterozygosity of the gene locus. Twenty-five cases were included (11 cases with and 14 cases without metastasis). Loss of heterozygosity for the nm-23 gene locus was seen in 7 of 22 (32%) informative tumors. Using immunohistochemistry, most tumors expressed nm-23, though decreased expression was seen in 10 of 25 (40%) cases. Only 2 tumors showed negative expression. We did not find a correlation between either protein expression or loss of heterozygosity with metastatic disease or any other adverse prognostic factors in this group of high-stage laryngeal squamous cell carcinomas. These data imply that nm-23 may be tumor suppressor gene involved in HNSCC but that it may not function as a tumor metastasis suppressor in high-stage laryngeal carcinoma.


Cognition, Health-Related Quality of Life, and Depression Ten Years after Moderate to Severe Traumatic Brain Injury: A Prospective Cohort Study.
The aim of this study was to evaluate cognitive function 10 years after moderate-severe traumatic brain injury (TBI) and to investigate the associations among cognitive function, depression, and health-related quality of life (HRQoL). In this prospective cohort study, with measurements at 3, 6, 12, 18, 24, 36, and 120 months post-TBI, patients 18-67 years of age (n = 113) with moderate-severe TBI were recruited. Main outcome measures were depression (Center for Epidemiologic Studies-Depression Scale [CES-D]), subjective cognitive functioning (Cognitive Failure Questionnaire [CFQ]), objective cognitive functioning, and HRQoL (Medical Outcomes Study 36-Item Short Form Health Survey [SF-36]). Fifty of the initial 113 patients completed the 10 year follow-up. Twenty percent showed symptoms of depression (CES-D ≥ 16). These patients had more psychiatric symptoms at hospital discharge (p = 0.048) and were more often referred to rehabilitation or nursing homes (p = 0.015) than non-depressed patients. Further, they also had significantly lower scores in six of the eight subdomains of the SF-36. The non-depressed patients had equivalent scores to those of the Dutch norm-population on all subdomains of the SF-36. Cognitive problems at hospital discharge were related with worse cognitive outcome 10 years post-TBI, but not with depression or HRQoL. Ten years after moderate-severe TBI, only weak associations (p < 0.05) between depression scores and two objective cognitive functioning scores were found. However, there were moderate associations (p < 0.01) among depression scores, HRQoL, and subjective cognitive functioning. Therefore, signaling and treatment of depressive symptoms after moderate-severe TBI may be of major importance for optimizing HRQoL in the long term. We did not find strong evidence for associations between depression and objective cognitive functioning in the long term post-TBI. Disease awareness and selective dropping out may play a role in long-term follow-up studies in moderate-severe TBI. More long-term research is needed in this field.