The Validity of Implicit Association Test (IAT) measures of sexual attraction to children: a meta-analysis.
The current study presents a quantitative review of the discriminative and convergent validity of Implicit Association Test (IAT) measures adapted to assess sexual interest in children. IAT measures were able to distinguish sex offenders against children (SOC) from non-SOC (M weighted d from random-effects = 0.63, 95 % CI [0.42-0.83], N = 707, k = 12). The largest group differences were found between SOC and non-offenders, followed by non-sex offenders and rapists. IAT measures using sex versus not sex (and similar attribute categories, such as sex vs. neutral) provided superior discrimination compared to IAT measures using sexy versus not sexy (and similar attribute categories, such as erotic vs. non-erotic). The IAT measures had a moderate relationship to self-report (r = .27, 95 % CI [.13-.40], N = 182), sexual offense history variables (r = .27, 95 % CI [.08-.43], N = 145), and viewing time (r = .30, 95 % CI [.16-.43], N = 180) measures of sexual interest in children. Although these IAT measures can discriminate between groups and show convergence with other measures of sexual interest, a better understanding of the construct validity of these tools is required before their use in the assessment, treatment, and supervision of sex offenders.


Presence of adenylate cyclase activity in Golgi and other fractions from rat liver. I. Biochemical determination.
The distribution of adenylate cyclase (AC) in Golgi and other cell fractions from rat liver was studied using the Golgi isolation procedure of Ehrenreich et al. In liver homogenate the AC activity was found to decay with time, but addition of 1 mM EGTA reduced the rate of enzyme loss. The incorporation of 1 mM EGTA into the sucrose medium used in the initial two centrifugal steps of the Golgi isolation method stabilized the enzyme activity throughout the entire procedure and resulted in good enzyme recovery. In such preparations, AC activity was demonstrated to be associated not only with plasma membranes but also with Golgi membranes and smooth microsomal membranes as well. Furthermore, under the conditions used, enzyme activity was also associated with the 105,000 g x 90 min supernatant fraction. The specific activity of the liver homogenate was found to be 2.9 pmol-mg protein-1-min-1, the nonsedimentabel and microsomal activity was of the same order of magnitude, but the Golgi and plasma membrane activities were much higher. The specific activity of plasma membrane AC was 29 pmol-mg proten-1-min-1. The Golgi activity varied in the three fractions, with the highest activity (14 pmol) in GF1 lowest activity (1.8) in GF2, and intermediate activity (5.5) in GF3, when the Golgi activity was corrected for the presence of content protein, the activity in GF1 became much higher (9 x) than that of the plasma membrane while the activities in GF2 and GF3 were comparable to that of plasma membrane. In all locations studied, the AC was sensitive to NaF stimulation, especially the enzyme associated with Golgi membranes. The activities in plasma and microsomal membranes were stimulated by glucagon, whereas the Golgi and nonsedimentable AC were not.


Homogeneous nucleation at high supersaturation and heterogeneous nucleation on microscopic wettable particles: A hybrid thermodynamic/density-functional theory.
Homogeneous nucleation at high supersaturation of vapor and heterogeneous nucleation on microscopic wettable particles are studied on the basis of Lennard-Jones model system. A hybrid classical thermodynamics and density-functional theory (DFT) approach is undertaken to treat the nucleation problems. Local-density approximation and weighted-density approximation are employed within the framework of DFT. Special attention is given to the disjoining pressure of small liquid droplets, which is dependent on the thickness of wetting film and radius of the wettable particle. Different contributions to the disjoining pressure are examined using both analytical estimations and numerical DFT calculation. It is shown that van der Waals interaction results in negative contribution to the disjoining pressure. The presence of wettable particles results in positive contribution to the disjoining pressure, which plays the key role in the heterogeneous nucleation. Several definitions of the surface tension of liquid droplets are discussed. Curvature dependence of the surface tension of small liquid droplets is computed. The important characteristics of nucleation, including the formation free energy of the droplet and nucleation barrier height, are obtained.


Catastrophic healthcare expenditure and impoverishment in tropical deltas: evidence from the Mekong Delta region.
Universal health coverage implies that people obtain the health services they need without experiencing financial hardship. While the factors contributing to catastrophic health expenditure (CHE) among households are well understood, few studies have examined this relationship in the context of environmentally vulnerable regions, such as tropical deltas. This study aims to examine the disparities in the prevalence of CHE and impoverishment due to out-of-pocket (OOP) healthcare payments in the Mekong Delta in comparison with rest of Vietnam. It also intends to investigate the associations between economic and environmental shocks, CHE and the impoverishment from healthcare payments. Using data from the Vietnam Household Living Standards Survey 2012, the prevalence of CHE was estimated from the fraction of healthcare costs in relation to household consumption expenditure. The poverty headcount was estimated using the total household consumption expenditure considering both with and without OOP expenditure for healthcare in comparison with the national poverty-line. Simple and multiple logistic regression models were used to examine the associations between geography, health systems, environmental and demographic variables and OOP healthcare expenditure related CHE, and impoverishment respectively. Both the level of OOP household healthcare expenditure and the proportion of households suffering from impoverishment as the result of such payments were higher in the Mekong Delta region compared to rest of Vietnam. Although the results from the multiple regression analysis showed that households in the Mekong Delta region were significantly less likely to suffer from CHE, they were significantly more likely to be impoverished due to OOP healthcare expenditure. While health insurance membership did not have a significant effect on either outcomes, households that faced an economic or an environmental shock in past 5 years were considerably more likely to suffer from CHE and impoverishment from OOP healthcare payments. The findings suggest that the financial protection capacity of health insurance schemes in Vietnam should be improved and expanded to reduce impoverishment as the result of OOP healthcare payments, particularly in the Mekong Delta region. Additional investments in disaster preparedness strategies can further help to reduce the financial burden of households in this environmentally vulnerable region.


Nonlinear absorption microscopy.
For the past two decades, nonlinear microscopy has been developed to overcome the scattering problem in thick tissue imaging. Owing to its increased imaging depth and high spatial resolution, nonlinear microscopy becomes the first choice for imaging living tissues. The use of nonlinear optical effects not only facilitates the signal originating from an extremely small volume defined by light focusing but also provides novel contrast mechanisms with molecular specificity. Nonlinear absorption is a nonlinear optical effect in which the absorption coefficient depends on excitation intensity. As a commonly used spectroscopy tool, nonlinear absorption measurement uncovers many photophysical and photochemical processes correlated with electronic states of molecules. Recently we have been focusing on adapting this spectroscopy method to a microscopy imaging technique. The effort leads to a novel modality in nonlinear microscopy-nonlinear absorption microscopy. This article summarizes the principles and instrumentation of this imaging technique and highlights some of the recent progress in applying it to imaging skin pigmentation and microvasculature under ex vivo or in vivo conditions.


Combined use of ultrasonography and contrast enhanced computed tomography to evaluate acute necrotizing pancreatitis in two dogs.
The imaging findings in two miniature schnauzers with acute necrotizing pancreatitis are described. Both dogs were treated previously for diabetes mellitus and hyperlipidemia. Vomiting, anorexia, and lethargy were observed in both dogs at presentation. Laboratory evaluations supportive of pancreatitis included left shift, abnormally high serum amylase and lipase activities, hypocalcemia, and abnormally high serum activities of liver enzymes. Sonographically, both dogs had diffusely enlarged hypoechoic pancreatic tissue with anechoic foci compatible with necrosis, abscessation, phlegmon, and pseudocysts formation. Contrast-enhanced computed tomography (CT) findings in both dogs were compatible with pancreatic necrosis. Dog 1 was managed medically for 11 days. Follow-up CT scan in this dog disclosed decreased pancreatic size and increased contrast enhancement compatible with partial resolution of pancreatitis.


The Impact o Molecular Techniques on the Study of Meningococcal Disease.
Neisseria meningitidis, the meningococcus, is normally a harmless commensal bacterium that colonises the naso/oropharynx of humans. This antigenically variable gram-negative diplococcus has the potential, however, to cause rapidly progressing meningitis and fulminant septicemia, either separately or together (1,2). Once present in the bloodstream, meningococci grow rapidly and their highly toxic lipo-oligosaccharides frequently cause extensive tissue damage and severe toxic shock. The progress of the disease is swift, and death often occurs within hours of the onset of symptoms (3). Even in countries where meningococcal infection is relatively rare, it remains a high priority for public health services because of the high mortality rates of fulminant septicemic disease (which can be up to 40% even when intensive supportive therapy is available), the high proportion of sequelae in patients who have recovered (including brain damage and digit or limb loss), and the age groups most susceptible (young children and, to a lesser extent, teenagers) (4,5).


How I treat patients with thrombotic thrombocytopenic purpura: 2010.
Thrombotic thrombocytopenic purpura (TTP) is the common name for adults with microangiopathic hemolytic anemia, thrombocytopenia, with or without neurologic or renal abnormalities, and without another etiology; children without renal failure are also described as TTP. The diagnosis of TTP is an indication for plasma exchange treatment, but beginning treatment requires sufficient confidence in the diagnosis to justify the risk of plasma exchange complications. Documentation of a severe deficiency of plasma ADAMTS13 activity, defined as less than 10% of normal, is not essential for the diagnosis of TTP. Some patients without severe ADAMTS13 deficiency may benefit from plasma exchange treatment; in addition, some patients with severe ADAMTS13 deficiency may subsequently be diagnosed with another cause for their clinical features. However, severe acquired ADAMTS13 deficiency does define a subgroup of patients who appear to benefit from treatment with corticosteroids and other immunosuppressive agents in addition to plasma exchange but who have a high risk for relapse. Approximately 80% of patients survive their acute episode, a survival rate that has not changed since the introduction of plasma exchange treatment. Although recovery may appear to be complete, many patients have persistent minor cognitive abnormalities. More effective as well as safer treatment for TTP is needed.


Size-dependent costs of migration: Migrant bird species are subordinate to residents, but only at small body sizes.
Migrant species are commonly thought to be poor competitors in aggressive interactions with resident species. However, no studies have tested whether this relationship is widespread. Here, we compare the behavioural dominance of closely related species of migratory and nonmigratory birds, testing whether migrants are consistently subordinate to resident species in aggressive contests. We compiled published behavioural dominance data involving migrant and resident congeners, gathering additional data on the body mass and migratory distance of each species. Focal species included a diverse array of birds (28 taxonomic families, 12 orders) from around the world. We found that migrant species are usually subordinate to resident species, but that this relationship disappears at larger body sizes. For smaller birds (<500 g), resident species were behaviourally dominant in 83%-88% of comparisons; for larger birds (>500 g), resident species were dominant in only 25%-30% of comparisons. The relative difference in body mass best predicted dominance relationships among species, with larger species dominant in 80%-84% of comparisons. When migrant and resident masses were equal, however, resident species were still more likely to be dominant in smaller birds, suggesting that other factors may also contribute to the subordinate status of migrants. Overall, our results suggest that in smaller species, the evolution of migration is associated with lighter weights and other traits that compromise the competitive abilities of migrants relative to residents. In contrast, larger species appear able to evolve migration without compromising their size or competitive abilities in aggressive contests, suggesting size-dependent constraints on the evolution of migration.


Effects of lipid-lowering therapy on reduction of cardiovascular events in patients with end-stage renal disease requiring hemodialysis.
In the general population, dyslipidemia is an established independent risk factor for cardiovascular disease. In patients with end-stage renal disease (ESRD), comorbid cardiovascular disease is present at alarming rates, and those who require hemodialysis and have cardiovascular disease continue to have a high mortality rate. Lipid abnormalities associated with chronic kidney disease (CKD) vary depending on the stage of disease (stages 1-5), but low-density lipoprotein cholesterol (LDL) has been established as the primary lipid treatment target. Guidelines support an LDL level of less than 100 mg/dl in patients with all stages of CKD, except when the triglyceride level is above 500 mg/dl. As patients progress to stage 5 CKD (ESRD with hemodialysis), the high triglyceride, low high-density lipoprotein cholesterol, and increased lipoprotein(a) levels of the early stages become more pronounced, with increases in small dense LDL particles; however, total cholesterol and LDL values remain normal or decrease. In patients undergoing hemodialysis, lipid abnormalities are driven by an increase in hepatic secretion and delayed catabolism of very low-density lipoproteins, as well as a reduction in lipoprotein lipase and hepatic lipase. Epidemiologic data support the role of cholesterol lowering as a means to lower cardiovascular events in the hemodialysis population. We conducted a literature search of various databases (1966-September 2009) to identify relevant clinical trials that evaluated the efficacy and safety of multiple lipid-lowering agents for the treatment of dyslipidemia in patients with ESRD requiring hemodialysis. Only those trials that used clinical primary end points of coronary heart disease (e.g., cardiovascular death, myocardial infarction, stroke) were included in this review. Evidence demonstrates that 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin) therapy (i.e., atorvastatin and rosuvastatin) significantly reduces surrogate cardiovascular markers, particularly LDL, in patients with ESRD requiring hemodialysis; however, no statin has proved to reduce cardiovascular morbidity or mortality in this population. Trials evaluating omega-3 fatty acids did not show significant reductions in LDL or cardiovascular events in this population. Clinicians should appreciate these limitations when deciding whether to continue lipid-lowering pharmacotherapy in these patients, depending on their overall cardiovascular risk assessment.


CDK9 Inhibition Induces a Metabolic Switch that Renders Prostate Cancer Cells Dependent on Fatty Acid Oxidation.
Cyclin-dependent kinase 9 (CDK9), a key regulator of RNA-polymerase II, is a candidate drug target for cancers driven by transcriptional deregulation. Here we report a multi-omics-profiling of prostate cancer cell responses to CDK9 inhibition to identify synthetic lethal interactions. These interactions were validated using live-cell imaging, mitochondrial flux-, viability- and cell death activation assays. We show that CDK9 inhibition induces acute metabolic stress in prostate cancer cells. This is manifested by a drastic down-regulation of mitochondrial oxidative phosphorylation, ATP depletion and induction of a rapid and sustained phosphorylation of AMP-activated protein kinase (AMPK), the key sensor of cellular energy homeostasis. We used metabolomics to demonstrate that inhibition of CDK9 leads to accumulation of acyl-carnitines, metabolic intermediates in fatty acid oxidation (FAO). Acyl-carnitines are produced by carnitine palmitoyltransferase enzymes 1 and 2 (CPT), and we used both genetic and pharmacological tools to show that inhibition of CPT-activity is synthetically lethal with CDK9 inhibition. To our knowledge this is the first report to show that CDK9 inhibition dramatically alters cancer cell metabolism.


Daily Satellite Observations of Methane from Oil and Gas Production Regions in the United States.
Production of oil and natural gas in North America is at an all-time high due to the development and use of horizontal drilling and hydraulic fracturing. Methane emissions associated with this industrial activity are a concern because of the contribution to climate radiative forcing. We present new measurements from the space-based TROPOspheric Monitoring Instrument (TROPOMI) launched in 2017 that show methane enhancements over production regions in the United States. In the Uintah Basin in Utah, TROPOMI methane columns correlated with in-situ measurements, and the highest columns were observed over the deepest parts of the basin, consistent with the accumulation of emissions underneath inversions. In the Permian Basin in Texas and New Mexico, methane columns showed maxima over regions with the highest natural gas production and were correlated with nitrogen-dioxide columns at a ratio that is consistent with results from in-situ airborne measurements. The improved detail provided by TROPOMI will likely enable the timely monitoring from space of methane emissions associated with oil and natural gas production.


Perceived neighborhood environment and park use as mediators of the effect of area socio-economic status on walking behaviors.
Access to local parks can affect walking levels. Neighborhood environment and park use may influence relationships between neighborhood socioeconomic status (SES) and walking. Self-report data on perceived park features, neighborhood environment, park use, neighborhood walking and sociodemographics were obtained from a sample of Australian adults, living in high/low SES areas. Surveys were mailed to 250 randomly selected households within 500m of 12 matched parks. Mediating effects of perceived environment attributes and park use on relationships between area-SES and walking were examined. Mean frequency of local park use was higher for high-SES residents (4.36 vs 3.16 times/wk, P < .01), who also reported higher levels of park safety, maintenance, attractiveness, opportunities for socialization, and neighborhood crime safety, aesthetics, and traffic safety. Safety and opportunity for socialization were independently positively related to monthly frequency of visits to a local park which, in turn, was positively associated with walking for recreation and total walking. Residents of higher SES areas reported an average 22% (95% CI: 5%, 37%) more weekly minutes of recreational walking than their low SES counterparts. Residents of high-SES areas live in environments that promote park use, which positively contributes to their weekly amounts of overall and recreational walking.


Delayed cervical palsy following cervical spine fusion leads to an increase in hospital-related costs.
Delayed cervical palsy (DCP) is a known complication following cervical spine surgery. While most DCPs eventually improve, they can result in significant temporary disability. Postoperative complications affect hospital length of stay (LOS) as well as overall hospital cost. The authors sought to determine the hospital cost of DCP after cervical spine fusion operations. A retrospective review of patients undergoing cervical fusion for degenerative disease at the Mayo Clinic from 2008 to 2012 was performed. Patients who developed DCPs not attributable to intraoperative trauma were included. All nonoperative-related costs were compared with similar costs in a control group matched according to age, sex, and surgical approach. All costs and services were reflective of the standard costs for the current year. Raw cost data were presented using ratios due to institutional policy against publishing cost data. There were 27 patients (18 men, 9 women) who underwent fusion and developed a DCP over the study period. These patients were compared with 24 controls (15 men, 9 women) undergoing fusion in the same time period. There was no difference between patients and controls in mean age (62.4 ± 3.1 years vs. 63.8 ± 2.5 years, respectively; p = 0.74), LOS (4.2 ± 3.3 days vs 3.8 ± 4.5 days, respectively; p = 0.43), or operating room-related costs (1.08 ± 0.09 vs. 1.0 ± 0.07, respectively; p = 0.58). There was a significant difference in nonoperative hospital-related costs between patients and controls (1.67 ± 0.15 vs 1.0 ± 0.09, respectively; p = 0.04). There was a significantly higher utilization of postoperative imaging (CT or MRI) in the DCP group (14/27, 52%) when compared with the matched cohort (4/24, 17%; p = 0.018), and a significantly higher utilization of physiatry services (24/27 [89%] vs 15/24 [63%], respectively; p = 0.046). While DCPs did not significantly prolong the length of hospitalization, they did increase hospital-related costs. This method could be further extrapolated to model costs of other complications as well.


cGAS-STING signalling in cancer: striking a balance with chromosomal instability.
Chromosomal instability (CIN) is a hallmark of cancer that drives tumour evolution. It is now recognised that CIN in cancer leads to the constitutive production of misplaced DNA in the form of micronuclei and chromatin bridges. These structures are detected by the nucleic acid sensor cGAS, leading to the production of the second messenger 2'3'-cGAMP and activation of the critical hub of innate immune signalling STING. Activation of this immune pathway should instigate the influx and activation of immune cells, resulting in the eradication of cancer cells. That this does not universally occur in the context of CIN remains an unanswered paradox in cancer. Instead, CIN-high cancers are notably adept at immune evasion and are highly metastatic with typically poor outcomes. In this review, we discuss the diverse facets of the cGAS-STING signalling pathway, including emerging roles in homeostatic processes and their intersection with genome stability regulation, its role as a driver of chronic pro-tumour inflammation, and crosstalk with the tumour microenvironment, which may collectively underlie its apparent maintenance in cancers. A better understanding of the mechanisms whereby this immune surveillance pathway is commandeered by chromosomally unstable cancers is critical to the identification of new vulnerabilities for therapeutic exploitation.


Montreal Cognitive Assessment (MoCA) for HIV-Associated Neurocognitive Disorders.
This study aims to systematically review the evidence on the accuracy of the Montreal Cognitive Assessment (MoCA) test for diagnosing HIV-associated neurocognitive disorders (HAND) and to outline the quality and quantity of research evidence available about the accuracy of MoCA in populations infected with HIV. We conducted a systematic literature review, searching five databases from inception until January 2019. We extracted dichotomized positive and negative test results at various thresholds and calculated the sensitivity and specificity of MoCA. Quality assessment was performed according to the Quality Assessment of Diagnostic Accuracy Studies 2 (QUADAS-2) criteria. Eight cross-sectional studies met the inclusion criteria for meta-analysis. Overall, 1014 patients were included but most studies recruited small samples. Recruitment period ranged from 2009 to 2015. We assessed most studies as being applicable to the review question though we had concerns about the selection of participants in three studies. The accuracy of MoCA for diagnosing HAND was reported at six cut-off points (scores 22-27). The MoCA test provides information about general cognitive functioning disturbances that contribute to a diagnosis of HAND. A lower threshold than the original cut-off of 26 is probably more useful for optimal screening of HAND, as it lowers false positive rates and improves diagnostic accuracy. Nonetheless, the choice of cut-off always comes with a sensitivity-specificity trade-off, the preferred cut point depending on whether sensitivity or specificity is more valuable in a given context.


White-Matter Structural Connectivity Underlying Human Laughter-Related Traits Processing.
Most research into the neural mechanisms of humor has not explicitly focused on the association between emotion and humor on the brain white matter networks mediating this connection. However, this connection is especially salient in gelotophobia (the fear of being laughed at), which is regarded as the presentation of humorlessness, and two related traits, gelotophilia (the enjoyment of being laughed at) and katagelasticism (the enjoyment of laughing at others). Here, we explored whether the topological properties of white matter networks can account for the individual differences in the laughter-related traits of 31 healthy adults. We observed a significant negative correlation between gelotophobia scores and the clustering coefficient, local efficiency and global efficiency, but a positive association between gelotophobia scores and path length in the brain's white matter network. Moreover, the current study revealed that with increasing individual fear of being laughed at, the linking efficiencies in superior frontal gyrus, anterior cingulate cortex, parahippocampal gyrus, and middle temporal gyrus decreased. However, there were no significant correlations between either gelotophilia or katagelasticism scores or the topological properties of the brain white matter network. These findings suggest that the fear of being laughed at is directly related to the level of local and global information processing of the brain network, which might provide new insights into the neural mechanisms of the humor information processing.


Nitrogen-containing bisphosphonates inhibit the mevalonate pathway and prevent post-translational prenylation of GTP-binding proteins, including Ras.
Bisphosphonates are currently the most important class of antiresorptive drugs used for the treatment of metabolic bone diseases. Although the molecular targets of bisphosphonates have not been identified, these compounds inhibit bone resorption by mechanisms that can lead to osteoclast apoptosis. Bisphosphonates also induce apoptosis in mouse J774 macrophages in vitro, probably by the same mechanisms that lead to osteoclast apoptosis. We have found that, in J774 macrophages, nitrogen-containing bisphosphonates (such as alendronate, ibandronate, and risedronate) inhibit post-translational modification (prenylation) of proteins, including the GTP-binding protein Ras, with farnesyl or geranylgeranyl isoprenoid groups. Clodronate did not inhibit protein prenylation. Mevastatin, an inhibitor of 3-hydroxy-3-methylglutatyl (HMG)-CoA reductase and hence the biosynthetic pathway required for the production of farnesyl pyrophosphate and geranylgeranyl pyrophosphate, also caused apoptosis in J774 macrophages and murine osteoclasts in vitro. Furthermore, alendronate-induced apoptosis, like mevastatin-induced apoptosis, could be suppressed in J774 cells by the addition of farnesyl pyrophosphate or geranylgeranyl pyrophosphate, while the effect of alendronate on osteoclast number and bone resorption in murine calvariae in vitro could be overcome by the addition of mevalonic acid. These observations suggest that nitrogen-containing bisphosphonate drugs cause apoptosis following inhibition of post-translational prenylation of proteins such as Ras. It is likely that these potent antiresorptive bisphosphonates also inhibit bone resorption by preventing protein prenylation in osteoclasts and that enzymes of the mevalonate pathway or prenyl protein transferases are the molecular targets of the nitrogen-containing bisphosphonates. Furthermore, the data support the view that clodronate acts by a different mechanism.


PRELIMINARY OBSERVATIONS ON A CASE OF PHYSIOLOGICAL ALBUMINURIA.
1. The proteids consisted of albumins and globulins, varying in quantity between the extremes of .9010 grm. and .2592 grm. in the 24 hours, but usually between .7 and .37 grm., the average being .5317 grm. 2. The quantity of the proteids varies directly as the urea, inversely as the external temperature. The relation between the quantity of urine and the quantity of proteid is not constant. The quantity of the proteida is little affected by diuretics; it tends to be increased by certain drugs that act upon the circulation. It is lessened during sleep. It is apt to suffer a sudden temporary increase, returning as suddenly to the usual average. For this phenomenon we have no explanation. 3. From .5449 to .6616 grm. of coagulable proteids a day (.0079 to .0094 grm. per kilo of body-weight) may be excreted through the urine for an indefinite time by an otherwise healthy individual without damage to either kidneys or organism.


Diversity in the dynamical behaviour of a compartmentalized programmable biochemical oscillator.
In vitro compartmentalization of biochemical reaction networks is a crucial step towards engineering artificial cell-scale devices and systems. At this scale the dynamics of molecular systems becomes stochastic, which introduces several engineering challenges and opportunities. Here we study a programmable transcriptional oscillator system that is compartmentalized into microemulsion droplets with volumes between 33 fl and 16 pl. Simultaneous measurement of large populations of droplets reveals major variations in the amplitude, frequency and damping of the oscillations. Variability increases for smaller droplets and depends on the operating point of the oscillator. Rather than reflecting the stochastic kinetics of the chemical reaction network itself, the variability can be attributed to the statistical variation of reactant concentrations created during their partitioning into droplets. We anticipate that robustness to partitioning variability will be a critical challenge for engineering cell-scale systems, and that highly parallel time-series acquisition from microemulsion droplets will become a key tool for characterization of stochastic circuit function.


Influence of chlorpromazine, bleomycin and WR-2721 singly or in combination on the formation of radiation-induced micronuclei in mice bone marrow.
Radiotherapy of tumors is often accompanied by inadvertant side effects on the normal tissues after completion of the treatment, which may later on result in the development of secondary neoplasia. Aim was to investigate the effect of combination of radiosensitizing agents in combination with a radioprotector on the frequency of micronuclei in mouse bone marrow and to find out whether the administration of combination of radiosensitizing agents in combination with a radioprotective drug before irradiation will offer any significant advantage over the drugs used singly. Female BALB/c mice were divided into 8 groups according to the treatment they received viz. 1. DDW (double distilled water), 2. chlorpromazine (CPZ) 10 mg/kg body-weight, 3. CPZ 15 mg/kg body-weight 4. bleomycin (BLM), 5. WR-2721, 6. CPZ (10 mg) + WR-2721, 7. CPZ (15 mg) + WR-2721, 8. BLM + WR-2721. After 30 min of drug/s administration the animals were exposed to either 0 or 4 Gy of 60Co g-radiation. The animals were killed at 24 h post-irradiation by cervical dislocation and the micronuclei were prepared according to the method described by Jagetia [17]. The administration of CPZ (10 or 15 mg) alone increased the frequency of micronuclei significantly when compared to the DDW treatment. The exposure of mice with 4 Gy resulted in a significant increase in the frequency of micronuclei compared to the concurrent control groups. The frequency of micronuclei increased significantly in CPZ (15 mg) and BLM + irradiated groups. However, treatment with WR-2721 before irradiation reduced the frequency of micronuclei by approximately 50% of the DDW + irradiated group. A further reduction in the frequency of micronuclei was observed when WR-2721 was combined with CPZ (10 and 15 mg) before irradiation. A combination of BLM with WR-2721 also resulted in a nonsignificant reduction in the frequency of micronuclei.


Microbial cell wall agents as an occupational hazard.
Organic dusts cause inflammatory reactions in the tissues exposed. The lung and the cells lining the surface of the respiratory tract are a primary target. Many receptors have been shown to react specifically on the presence of microorganisms that are ubiquitous elements in organic dusts. There is a great variability in the individual response to organic dusts. Almost 50% of Caucasians are hyporesponders to LPS exposure, and people with alpha-1-antitrypsin deficiency are hyperresponsive to organic dust exposure. The diseases resulting from organic dust exposures include asthma, allergy, hypersensitivity pneumonitis and toxic pneumonitis (organic dust toxic syndrome). This paper deals with inflammation and the subsequent mechanism of disease as it is encountered in industries with these exposures. Toxicological studies including human experimental exposures and ex vivo studies of cells are described. Cellular reactions are mediated through the attachment of, e.g. LPS and beta (1,3)-D-glucan to lipopolysaccharide binding protein, CD14 and Toll-like receptors. The relation between protein release and the gene activation is described. Furthermore, studies of the individual susceptibility will be reviewed.


Hemiarthroplasty and total hip arthroplasty in 30,830 patients with hip fractures: data from the Dutch Arthroplasty Register on revision and risk factors for revision.
Background and purpose - In the Netherlands about 40% of hip fractures are treated with a hemiarthroplasty (HA) or a total hip arthroplasty (THA). Although these procedures are claimed to have fewer complications than osteosynthesis (i.e., reoperation), complications still occur. Analyses of data from national registries with adequate completeness of revision surgery are important to establish guidelines to diminish the risk for revision. We identified risk factors for revision. Patients and methods - All patients older than 50 years of age with a hip fracture treated with arthroplasty by orthopedic surgeons and registered in the (national) Dutch arthroplasty register (LROI) were included in the study. In this register, patient characteristics and surgical details were prospectively collected. Revision surgery and reasons for revision were evaluated. A proportional hazard ratio model for revision was created using competing risk analysis (with death as competing risk). Results - 1-year revision rate of HA was (cumulative incidence function [CIF] 1.6% (95% CI 1.4-1.8) and THA 2.4% (CI 2.0-2.7). Dislocation was the most common reason for revision in both groups (HA 29%, THA 41%). Male sex, age under 80 years, posterolateral approach, and uncemented stem fixation were risk factors for revision in both THA and HA. THA patients with ASA classification III/IV were revised more often, whereas revision in the HA cohort was performed more often in ASA I/II patients. Interpretation - After arthroplasty of hip fractures, both a posterolateral approach and an uncemented hip stem have higher risks for revision surgery compared with an anterolateral approach and an cemented stem.


User-centered design and the development of patient decision aids: protocol for a systematic review.
Providing patient-centered care requires that patients partner in their personal health-care decisions to the full extent desired. Patient decision aids facilitate processes of shared decision-making between patients and their clinicians by presenting relevant scientific information in balanced, understandable ways, helping clarify patients' goals, and guiding decision-making processes. Although international standards stipulate that patients and clinicians should be involved in decision aid development, little is known about how such involvement currently occurs, let alone best practices. This systematic review consisting of three interlinked subreviews seeks to describe current practices of user involvement in the development of patient decision aids, compare these to practices of user-centered design, and identify promising strategies. A research team that includes patient and clinician representatives, decision aid developers, and systematic review method experts will guide this review according to the Cochrane Handbook and PRISMA reporting guidelines. A medical librarian will hand search key references and use a peer-reviewed search strategy to search MEDLINE, EMBASE, PubMed, Web of Science, the Cochrane Library, the ACM library, IEEE Xplore, and Google Scholar. We will identify articles across all languages and years describing the development or evaluation of a patient decision aid, or the application of user-centered design or human-centered design to tools intended for patient use. Two independent reviewers will assess article eligibility and extract data into a matrix using a structured pilot-tested form based on a conceptual framework of user-centered design. We will synthesize evidence to describe how research teams have included users in their development process and compare these practices to user-centered design methods. If data permit, we will develop a measure of the user-centeredness of development processes and identify practices that are likely to be optimal. This systematic review will provide evidence of current practices to inform approaches for involving patients and other stakeholders in the development of patient decision aids. We anticipate that the results will help move towards the establishment of best practices for the development of patient-centered tools and, in turn, help improve the experiences of people who face difficult health decisions. PROSPERO CRD42014013241.


Tumor necrosis factor-alpha induces vascular endothelial growth factor-C expression in rheumatoid synoviocytes.
To determine the expression of vascular endothelial growth factor-C (VEGF-C) in the synovial fluid of patients with rheumatoid arthritis (RA) and to investigate the regulation of VEGF-C production by major proinflammatory cytokines in fibroblast-like synoviocytes (FLS). The concentrations of VEGF-C, tumor necrosis factor-alpha (TNF-alpha), and interleukin 1beta (IL-1beta) were measured using an ELISA method in synovial fluids obtained from 20 patients with RA and 20 with osteoarthritis (OA). Primary cultured RA FLS were stimulated with TNF-alpha or IL-1beta, and the expression levels of VEGF-C mRNA and protein were assessed by quantitative real-time polymerase chain reaction and ELISA. Significantly higher levels of VEGF-C were found in RA synovial fluids compared to OA synovial fluids. VEGF-C levels showed a highly significant correlation with the levels of both TNF-alpha and IL-1beta in the synovial fluid of patients with RA. TNF-alpha stimulation significantly increased VEGF-C mRNA and protein expression in RA FLS in a dose-dependent manner. A tendency to increased expression of VEGF-C was also observed after IL-1beta stimulation in FLS. Overexpression of VEGF-C in FLS by stimulation with TNF-alpha may play an important role in the progression of synovial inflammation and hyperplasia in RA by contributing to local lymphangiogenesis and angiogenesis.


11Beta-hydroxysteroid dehydrogenase type 1 in human disease: a novel therapeutic target.
Patients with cortisol excess, Cushing's syndrome, develop a classical phenotype characterized by central obesity, hypertension, and increased cardiovascular mortality. Whilst this observation points to the importance of glucocorticoids, circulating cortisol excess is rare and does not explain the pathogenesis of many common conditions. At a tissue specific level, the enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) locally regenerates active cortisol from inactive cortisone amplifying glucocorticoid receptor activation in the context of normal circulating cortisol levels. Increased 11beta-HSD1 activity and expression have been implicated in the pathogenesis of many common conditions including, obesity, insulin resistance, the metabolic syndrome, polycystic ovarian syndrome, osteoporosis and glaucoma. Furthermore, selective 11beta-HSD1 inhibition has been proposed as a novel therapeutic strategy in many of these conditions. Here we review the role of 11beta-HSD1 in human disease and discuss the impact of selective 11beta-HSD1 inhibition.


Identification of an immunodominant peptide of HER-2/neu protooncogene recognized by ovarian tumor-specific cytotoxic T lymphocyte lines.
Synthetic peptide analogues of sequences in the HER-2 protooncogene (HER-2) were selected based on the presence of HLA-A2.1 anchor motifs to identify the epitopes on HER-2 recognized by ovarian tumor-reactive CTL. 19 synthetic peptides were evaluated for recognition by four HLA-A2 ovarian-specific cytotoxic T lymphocyte (CTL) lines obtained from leukocytes associated with ovarian tumors. The nonapeptide E75 (HER-2, 369-377:KIFGSLAFL) was efficient in sensitizing T2 cells for lysis by all four CTL lines. This peptide was specifically recognized by cloned CD8+ CTL isolated from one of the ovarian-specific CTL lines. E75-pulsed T2 cells inhibited lysis by the same CTL clone of both an HLA-A2+ HER-2high ovarian tumor and a HER-2high cloned ovarian tumor line transfected with HLA-A2, suggesting that this or a structurally similar epitope may be specifically recognized by these CTL on ovarian tumors. Several other HER-2 peptides were recognized preferentially by one or two CTL lines, suggesting that both common and private HER-2 epitopes may be immunogenic in patients with ovarian tumors. Since HER-2 is a self-antigen, these peptides may be useful for understanding mechanisms of tumor recognition by T cells, immunological tolerance to tumor, and structural characterization of tumor antigens.


Informed consent for biorepositories: assessing prospective participants' understanding and opinions.
Obtaining informed consent for the collection, storage, and future research use of biospecimens is challenging, as potentially complex and controversial information must be communicated clearly. We gathered input on a consent template developed for the Duke Biorepository from individuals representative of those who might one day consider contributing specimens. Forty subjects were recruited from the Durham, NC area and screened to achieve diversity by race/ethnicity, education, age, and sex. Cognitive interviews assessed participants' (a) understanding of information in the template, and (b) opinions about that information. Participants also completed a survey assessing trust in medical researchers. Interviewees seemed to understand the template. Although responses were diverse, majority views emerged: more than half were comfortable with indefinite biospecimen storage, periodic contact to update information and to inform participants of additional research opportunities, the prospect that commercial products could be developed, and the fact that profits would not be shared. More than half were willing to provide medical record access, although this was a primary concern for others. More than two thirds were comfortable with not receiving individual research results as a matter of routine, but many thought they should be informed of findings with serious health implications. Lack of trust in researchers was associated with declining certain consent options. Protecting and promoting trust in research is essential to fostering widespread participation in biorepositories. Biorepositories should also devise ways to communicate clearly about the research being conducted and what is being learned.


Iconic gesture in normal language and word searching conditions: a case of conduction aphasia.
Although there is a substantive body of research about the language used by individuals with aphasia, relatively little is known about their spontaneous iconic gesture. A single case study of LT, an individual with conduction aphasia indicated qualitative differences between the spontaneous iconic gestures produced alongside fluent speech and in tip of the tongue states. The current study examined the iconic gestures produced by another individual with conduction aphasia, WT, and a group of 11 control participants. Comparisons were made between iconic gestures produced alongside normal language and those produced alongside word-searching behaviour. Participants recounted the Tweety and Sylvester cartoon Canary Row. All gesture produced was analysed qualitatively and quantitatively. WT produced more iconic gestures than controls accompanying word searching behaviour, whereas he produced a similar frequency of iconic gestures to control participants alongside normal language. The iconic gestures produced in the two language contexts also differed qualitatively. Frequency of iconic gesture production was not affected by limb apraxia. This study suggests that there are differences between iconic gestures that are produced alongside normal language and those produced alongside word-searching behaviour. Theoretical and clinical implications of these findings are discussed.


[Nutritional needs of the preterm infant after hospital discharge].
The authors report their experience in the Division of Neonatology of the Catholic University of Rome about the choice of milk alimentation and mineral and vitamin supplementation before discharge and during the subsequent follow-up, with particular reference to very low-birthweight preterm infants (< 1500 g). Basing on empirical experiences, the authors emphasize the importance in current practice of post-conceptional age, with special regard to the kind of milk to choose after discharge and the time and terms of the weaning. Furthermore they stress nutritional, immuno-allergic and psychological advantages of human milk before and after hospital discharge, particularly related to the presence of long-chain polyunsaturated fatty acid (LCP), recently known to be essential on retina and brain development in the preterm infant. When breast milk is not available, the authors confirm the efficacy, before discharge, of preterm infant formulas and subsequently of infant formulas and after of follow-up formulas. The authors hope that the directions proposed by the American Academy of Pediatrics in 1983 will be modified in order to recommend cow-milk only after the first year of life of the infant. They finally suggest specific mineral and vitamin supplementations (iron, calcium, phosphorus, fluoride; vitamins K, D, E and folic acid), to be started after hospital discharge.


The pore structure of Clostridium perfringens epsilon toxin.
Epsilon toxin (Etx), a potent pore forming toxin (PFT) produced by Clostridium perfringens, is responsible for the pathogenesis of enterotoxaemia of ruminants and has been suggested to play a role in multiple sclerosis in humans. Etx is a member of the aerolysin family of β-PFTs (aβ-PFTs). While the Etx soluble monomer structure was solved in 2004, Etx pore structure has remained elusive due to the difficulty of isolating the pore complex. Here we show the cryo-electron microscopy structure of Etx pore assembled on the membrane of susceptible cells. The pore structure explains important mutant phenotypes and suggests that the double β-barrel, a common feature of the aβ-PFTs, may be an important structural element in driving efficient pore formation. These insights provide the framework for the development of novel therapeutics to prevent human and animal infections, and are relevant for nano-biotechnology applications.


Genomic prediction of residual feed intake in US Holstein dairy cattle.
Genomic selection is an important tool to introduce feed efficiency into dairy cattle breeding. The goals of the current research are to estimate genomic breeding values of residual feed intake (RFI) and to assess the prediction reliability for RFI in the US Holstein population. The RFI data were collected from 4,823 lactations of 3,947 Holstein cows in 9 research herds in the United States, and were pre-adjusted to remove phenotypic correlations with milk energy, metabolic body weight, body weight change, and for several environmental effects. In the current analyses, genomic predicted transmitting abilities of milk energy and of body weight composite were included into the RFI model to further remove the genetic correlations that remained between RFI and these energy sinks. In the first part of the analyses, a national genomic evaluation for RFI was conducted for all the Holsteins in the national database using a standard multi-step genomic evaluation method and 60,671 SNP list. In the second part of the study, a single-step genomic prediction method was applied to estimate genomic breeding values of RFI for all cows with phenotypes, 5,252 elite young bulls, 4,029 young heifers, as well as their ancestors in the pedigree, using a high-density genotype chip. Theoretical prediction reliabilities were calculated for all the studied animals in the single-step genomic prediction by direct inversion of the mixed model equations. In the results, breeding values were estimated for 1.6 million genotyped Holsteins and 60 million ungenotyped Holsteins, The genomic predicted transmitting ability correlations between RFI and other traits in the index (e.g., fertility) are generally low, indicating minor correlated responses on other index traits when selecting for RFI. Genomic prediction reliabilities for RFI averaged 34% for all phenotyped animals and 13% for all 1.6 million genotyped animals. Including genomic information increased the prediction reliabilities for RFI compared with using only pedigree information. All bulls had low reliabilities, and averaged to only 16% for the top 100 net merit progeny-tested bulls. Analyses using single-step genomic prediction and high-density genotypes gave similar results to those obtained from the national evaluation. The average theoretical reliability for RFI was 18% among the elite young bulls under 5 yr old, being lower in the younger generations of elite bulls compared with older bulls. To conclude, the size of the reference population and its relationship to the predicted population remain as the limiting factors in the genomic prediction for RFI. Continued collection of feed intake data is necessary so that reliabilities can be maintained due to close relationships of phenotyped animals with breeding stock. Considering the currently low prediction reliability and high cost of data collection, focusing RFI data collection on relatives of elite bulls that will have the greatest genetic contribution to the next generation will give more gains and profit.


Effects of caffeine on anterior pituitary and thyroid function in the rat.
We studied the effects of acute, intraperitoneal administration of caffeine on serum thyrotropin (TSH), growth hormone, prolactin, thyroxine and 3,3',5-triiodothyronine in rats. Caffeine lowered serum TSH and GH in a dose-dependent manner with ED50 values of 30 and approximately 50 mg/kg, respectively. TSH levels were depressed 1 to 6 hr after injection and correlated with serum caffeine levels greater than 20 micrograms/ml. The decrease in serum TSH was followed by decreases in serum 3,3',5-triiodothyroxine and thyroxine 4 hr after caffeine administration. Theophylline and theobromine had effects similar to those of caffeine on hormone levels. Caffeine did not significantly affect hormone secretion when incubated directly with rat pituitaries in vitro. Administration of antisomatostatin antiserum to rats blocked the inhibitory effects of caffeine on serum GH levels, suggesting that caffeine inhibits GH and TSH secretion by releasing hypothalamic somatostatin.


Routine pulmonary function tests in young adolescents with asthma in general practice.
To assess the value of performing routine pulmonary function tests by flow-loop spirometry in young adolescents with asthma. A prospective clinical study comparing clinical assessment and patients' self-reporting of asthma severity with the results of pulmonary function tests. General practice in a small rural community of about 30,000 people. Young adolescents with asthma, aged 10-15 years, were enrolled in the study over a two-year period from July 1993 to June 1995 when they presented for either elective, interval assessments or with an acute exacerbation of asthma. Discrepancy between (i) the doctor's and the patient's perception of asthma control (six scale measures) and the consequent management plans, and (ii) the results of pulmonary function tests that indicated less than adequate airway function (i.e., forced expiratory volume in one second as a percentage of predicted vital capacity for height and sex [FEV1%] less than 65% or average flow rate over the middle 50% of forced vital capacity as a percentage of predicted normal value [FEF25%-75%] less than 65%). Twenty-seven adolescents with asthma were assessed on a total of 37 occasions. The results of pulmonary function tests did not correlate with asthma symptoms and treatment in 11 of the 37 assessments (30%; 95% confidence interval [CI], 16%-47%). The 11 assessments were performed on eight patients. This small community-based study of adolescents with asthma supports the view that pulmonary function testing by flow-loop spirometry should be part of the routine assessment of acute and chronic asthmatics. Further study in a larger community is needed to clarify the frequency of over- and underestimation of asthma severity in this difficult age group.


Stillbirths: progress and unfinished business.
This first paper of the Lancet Series on ending preventable stillbirths reviews progress in essential areas, identified in the 2011 call to action for stillbirth prevention, to inform the integrated post-2015 agenda for maternal and newborn health. Worldwide attention to babies who die in stillbirth is rapidly increasing, from integration within the new Global Strategy for Women's, Children's and Adolescents' Health, to country policies inspired by the Every Newborn Action Plan. Supportive new guidance and metrics including stillbirth as a core health indicator and measure of quality of care are emerging. Prenatal health is a crucial biological foundation to life-long health. A key priority is to integrate action for prenatal health within the continuum of care for maternal and newborn health. Still, specific actions for stillbirths are needed for advocacy, policy formulation, monitoring, and research, including improvement in the dearth of data for effective coverage of proven interventions for prenatal survival. Strong leadership is needed worldwide and in countries. Institutions with a mandate to lead global efforts for mothers and their babies must assert their leadership to reduce stillbirths by promoting healthy and safe pregnancies.


A simple and quantitative liquid culture system to measure megakaryocyte growth using highly purified CFU-MK.
A new and quantitative liquid culture system has been developed to measure the production of megakaryocytes from megakaryocyte progenitor cells (colony-forming units-megakaryocyte [CFU-MK]). The system uses as a target population a glycoprotein (Gp) IIb/IIIa+ subpopulation of rat bone marrow cells previously demonstrated to be highly enriched for CFU-MK. GpIIb/IIIa+ cells were cultured at 5 x 10(4) cells/mL (10(4) cells/well) with test samples in 96-well tissue culture plates for 4 days at 37 degrees C. During the final 3 hours of incubation, the cells were pulsed with [14C]5-hydroxytryptamine creatinine sulfate (14C-serotonin). After incubation, the plates were washed and the cell pellets were lysed with Triton-X 100. The cell lysate was infiltrated into a commercially available solid scintillator and dried, and radioactivity was measured. In this assay system, rat interleukin-3 (IL-3) was found to be the most potent among known cytokines tested. Murine granulocyte-macrophage colony-stimulating factor (GM-CSF), human erythropoietin (Epo), human IL-6, and murine stem cell factor (SCF) each alone stimulated megakaryocyte growth but were much less active than rat IL-3. Plasma of rats rendered thrombocytopenic by injection of monoclonal antirat platelet GpIIb/IIIa antibody exhibited significant activity, and the active protein fractions partially purified from the plasma showed much higher activity, but normal rat plasma had no effect. This liquid culture system allows the measurement of a large number of test samples--including a wide variety of cytokines and unknown growth factors, alone or in combinations--and provides a simple method for evaluating the early proliferative events involving CFU-MK in the megakaryocyte differentiation pathway.


Common founder effects of hereditary hemochromatosis, Wilson´s disease, the long QT syndrome and autosomal recessive deafness caused by two novel mutations in the WHRN and TMC1 genes.
Genealogy and molecular genetic studies of a Swedish river valley population resulted in a large pedigree, showing that the hereditary hemochromatosis (HH) HFE/p.C282Y mutation is inherited with other recessive disorders such as Wilson´s disease (WND), a rare recessive disorder of copper overload. The population also contain individuals with the Swedish long QT syndrome (LQTS1) founder mutation (KCNQ1/p.Y111C) which in homozygotes causes the Jervell & Lange Nielsen syndrome (JLNS) and hearing loss (HL).Aims of the study were to test whether the Swedish long QT founder mutation originated in an ancestral HFE family and if carriers had an increased risk for hemochromatosis (HH), a treatable disorder. We also aimed to identify the pathogenic mutation causing the hearing loss disorder segregating in the pedigree. LQTS patients were asked about their ancestry and possible origin in a HH family. They were also offered a predictive testing for the HFE genotype. Church books were screened for families with hearing loss. One HH family had two members with hearing loss, who underwent molecular genetic analysis of the LQTS founder mutation, connexin 26 and thereafter exome sequencing. Another family with hearing loss in repeat generations was also analyzed for connexin 26 and underwent exome sequencing. Of nine LQTS patients studied, four carried a HFE mutation (two p.C282Y, two p.H63D), none was homozygous. Three LQTS patients confirmed origin in a female founder ( b 1694, identical to AJ b 1694, a HFE pedigree member from the Fax river. Her descent of 44 HH families, included also 29 families with hearing loss (HL) suggesting JLNS. Eleven LQTS probands confirmed origin in a second founder couple (b 1614/1605) in which the woman b 1605 was identical to a HFE pedigree member from the Fjällsjö river. In her descent there were not only 64 HH, six WND families, one JLNS, but also 48 hearing loss families. Most hearing loss was non syndromic and caused by founder effects of the late 16th century. One was of Swedish origin carrying the WHRN, c.1977delC, (p.S660Afs*30) mutation, the other was a TMC1(NM_138691),c.1814T>C,(p.L605P) mutation, possibly of Finnish origin. Deep human HFE genealogies show HFE to be associated with other genetic disorders like Wilson´s disease, LQTS, JLNS, and autosomal recessive hearing loss. Two new homozygous HL mutations in WHRN/p.S660Afs*30 and TMC1/p.L605P were identified,none of them previously reported from Scandinavia. The rarity of JLNS was possibly caused by miscarriage or intrauterine death. Most hearing loss (81.7%) was seen after 1844 when first cousin marriages were permitted. However, only 10 (10.3%) came from 1st cousin unions and only 2 (2.0 %) was born out of wedlock.


Noncovalent complexation of glucosylthioureidocalix[4]arenes with carboxylates and their gas-phase characteristics: an ESI-FTICR mass spectrometric study.
The noncovalent complexation of three glucosylcalix[4]arenes (1-3) towards 23 mono- and dicarboxylic acid anions were studied by ESI-FTICR mass spectrometry. Competitive complexation, collision-induced dissociation and gas-phase H/D-exchange experiments were performed to obtain information on selectivity of calixarenes towards carboxylates and characteristics of their complexes. The flexibility and number of glucose units of the host and the spatial disposition of the hydrogen bonding groups on the carboxylate guests were found to affect the selectivity of complexation strongly. The glucosylcalixarenes exhibited particular selectivity for dicarboxylic acid anions incorporating π-systems, and clear isomeric selectivity was observed for isophthalic among phthalic acid anions and for fumaric acid over maleic acid anion.


Calpain 3 cleaves filamin C and regulates its ability to interact with gamma- and delta-sarcoglycans.
Calpain 3 (C3) is the only muscle-specific member of the calcium-dependent protease family. Although neither its physiological function nor its in vivo substrates are known, C3 must be an important protein for normal muscle function as mutations in the C3 gene result in limb-girdle muscular dystrophy type 2A. Previous reports have shown that the ubiquitous calpains (mu and m) proteolyze filamins in nonmuscle cells. This observation suggests that the muscle-specific filamin C (FLNC) is a good candidate substrate for C3. Binding studies using recombinant proteins establish that recombinant C3 and native FLNC can interact. When these two proteins are translated in vitro and incubated together, C3 cleaves the C-terminal portion of FLNC. Cleavage is specific as C3 fails to cleave FLNC lacking its C-terminal hinge and putative dimerization domains. Cotransfection experiments in COS-7 cells confirm that C3 can cleave the C-terminus of FLNC in live cells. The C-terminus of FLNC has been shown to bind the cytoplasmic domains of both delta- and gamma-sarcoglycan. Removal of the last 127 amino acids from FLNC, a protein that mimics FLNC after C3 cleavage, abolishes this interaction with the sarcoglycans. These studies confirm that C3 can cleave FLNC in vitro and suggest that FLNC may be an in vivo substrate for C3, functioning to regulate protein-protein interactions with the sarcoglycans. Thus, calpain-mediated remodeling of cytoskeletal-membrane interactions, such as those that occur during myoblast fusion and muscle repair, may involve regulation of FLNC-sarcoglycan interactions.


Assessment of urinary mephenytoin metrics to phenotype for CYP2C19 and CYP2B6 activity.
(S)-Mephenytoin is selectively metabolised to (S)-4'-hydroxymephenytoin by CYP2C19. The urinary excretion of 4'-hydroxymephenytoin reflects the activity of individual enzymes. We evaluated fractioned urinary collection and beta-glucuronidase pre-treatment in order to determine the optimal CYP2C19 metrics. We also assessed whether urinary excretion of N-desmethylmephenytoin (nirvanol) might be a useful CYP2B6 metric in in vivo studies. A 50-mg dose of mephenytoin was administered to 52 volunteers as a component of phenotyping cocktails in four separate studies. Urine was collected up to 166 h post-dose. Urinary excretion of 4'-hydroxymephenytoin and nirvanol was quantified by liquid chromatography-tandem mass spectrometry, and common CYP2C19 and CYP2B6 genotypes were determined. Cumulative excretion of 4'-hydroxymephenytoin in urine with beta-glucuronidase treatment collected from before mephenytoin administration up to 12-16 h thereafter showed the greatest difference between CYP2C19 genotypes and the lowest intra-individual variability (7%). Renal elimination of nirvanol was highest for a *4/*4 individual and lowest for individuals carrying the *5/*5 and *1/*7 genotype, but lasted for several weeks, thus making its use in cross-over studies difficult. Cumulative urinary excretion of 4'-hydroxymephenytoin 0-12 h post-administration is a sensitive and reproducible metric of CYP2C19 activity, enabling the effect of a drug on CYP2C19 to be assessed in a small sample size of n=6 volunteers. While nirvanol excretion may reflect CYP2B6 activity in vivo, it is not useful for CYP2B6 phenotyping.


Effects of pupillary responses to luminance and attention on visual spatial discrimination.
The optic quality of the eyes is, at least in part, determined by pupil size. Large pupils let more light enter the eyes, but degrade the point spread function, and thus the spatial resolution that can be achieved (Campbell & Gregory, 1960). In natural conditions, the pupil is mainly driven by the luminance (and possibly the color and contrast) at the gazed location, but is also modulated by attention and cognitive factors. Whether changes in eyes' optics related to pupil size modulation by luminance and attention impacts visual processing was assessed in two experiments. In Experiment 1, we measured pupil size using a constantly visible display made of four disks with different luminance levels, with no other task than fixating the disks in succession. The results confirmed that pupil size depends on the luminance of the gazed stimulus. Experiment 2, using similar settings as Experiment 1, used a two-interval forced-choice design to test whether discriminating high spatial frequencies that requires covert attention to parafoveal stimuli is better during the fixation of bright disks that entails a small pupil size, and hence better eyes' optics, as compared to fixating dark disks that entails a large pupil size, and hence poorer eyes' optics. As in Experiment 1, we observed large modulations of pupil size depending on the luminance of the gazed stimulus, but pupil dynamics was more variable, with marked pupil dilation during stimulus encoding, presumably because the demanding spatial frequency discrimination task engaged attention. However, discrimination performance and mean pupil size were not correlated. Despite this lack of correlation, the slopes of pupil dilation during stimulus encoding were correlated to performance, while the slopes of pupil dilation during decision-making were not. We discuss these results regarding the possible functional roles of pupil size modulations.


Accelerating cancer systems biology research through Semantic Web technology.
Cancer systems biology is an interdisciplinary, rapidly expanding research field in which collaborations are a critical means to advance the field. Yet the prevalent database technologies often isolate data rather than making it easily accessible. The Semantic Web has the potential to help facilitate web-based collaborative cancer research by presenting data in a manner that is self-descriptive, human and machine readable, and easily sharable. We have created a semantically linked online Digital Model Repository (DMR) for storing, managing, executing, annotating, and sharing computational cancer models. Within the DMR, distributed, multidisciplinary, and inter-organizational teams can collaborate on projects, without forfeiting intellectual property. This is achieved by the introduction of a new stakeholder to the collaboration workflow, the institutional licensing officer, part of the Technology Transfer Office. Furthermore, the DMR has achieved silver level compatibility with the National Cancer Institute's caBIG, so users can interact with the DMR not only through a web browser but also through a semantically annotated and secure web service. We also discuss the technology behind the DMR leveraging the Semantic Web, ontologies, and grid computing to provide secure inter-institutional collaboration on cancer modeling projects, online grid-based execution of shared models, and the collaboration workflow protecting researchers' intellectual property.


Primary mineralization at the surfaces of implants.
Osteogenesis around implants is affected by the physical and chemical characteristics of the biomaterials used. The osteoprogenitor cells must migrate to the implant site and synthesize and secrete a mineralizable extracellular matrix. Because this is neo-bone formation, the mechanism by which the cells calcify their matrix involves extracellular organelles called matrix vesicles in a process termed "primary mineralization". Two different methods for assessing the effects of implant materials on primary mineralization are presented in this report. In the first approach, different implant materials used in dentistry and orthopedic surgery were placed in rat tibial bones after marrow ablation. Two groups of implants were used, bone-bonding and non-bonding materials. We examined the effects of the materials on calcification morphometrically by quantitating changes in matrix vesicle morphology and distribution in endosteal tissue around implants as compared with normal endosteal bone healing. In addition, matrix vesicles were isolated from the endosteal tissue around the implant as well as from the contralateral limb and were examined biochemically. The results demonstrated that bone-bonding materials induced a greater increase in matrix vesicle enzyme activity than did non-bonding materials. However, all materials caused changes in matrix vesicles that were different from those seen in normal endosteal bone formation following injury. The effects of implant materials on biochemical markers of mineralization, including specific activities of matrix vesicle alkaline phosphatase and phospholipase A2 and phosphatidylserine content, demonstrated a high correlation with the morphometric observations with regard to enhancement and/or delay of primary mineralization. In the other approach, we used a radioisotopic method to evaluate the effects of implant materials on primary mineralization. This analysis revealed that implants alter bone healing, as shown by the differential uptake of 99mTc and 32P in different bone compartments. Decreased 32P uptake by the organic phase in the presence of bone-bonding implants suggests that cleavage of 99mTcMD32P into its technetium and methylene diphosphonate moieties was inhibited by the presence of the implants. In summary, these approaches to evaluating the effects of materials on primary mineralization demonstrate that the marrow ablation model can easily distinguish between bone-bonding and non-bonding materials. The use of this model can be valuable in the development of new materials.


EULAR/EFORT recommendations for the diagnosis and initial management of patients with acute or recent onset swelling of the knee.
The European League Against Rheumatism (EULAR) and the European Federation of National Associations of Orthopaedics and Traumatology (EFORT) have recognised the importance of variation in diagnostic and therapeutic performance across disciplines, have found consensus in starting task forces aiming at achieving diagnostic and therapeutic uniformity, and have identified medical conditions with which representatives of both organisations will frequently be confronted in common clinical practice. The aim of the present work was to establish recommendations for the diagnosis and initial management of patients presenting with acute or recent onset swelling of the knee. The EULAR standard operating procedures for the elaboration and implementation of evidence-based recommendations were followed. In all, 11 rheumatologists from 11 countries and 12 orthopaedic surgeons from 7 countries met twice under the leadership of 2 conveners, a clinical epidemiologist and a research fellow. After carefully defining the content and procedures of the task force, research questions were developed, a comprehensive literature search was performed and the results were presented to the entire committee. Subsequently, a set of 10 recommendations was formulated based on evidence from the literature if available, and after discussion and consensus building. This is the first combined interdisciplinary project of rheumatologists and orthopaedic surgeons, successfully aiming at achieving consensus in the diagnosis and initial management of patients presenting with acute or recent onset swelling of the knee.


Holding the Hunger Games Hostage at the Gym: An Evaluation of Temptation Bundling.
We introduce and evaluate the effectiveness of temptation bundling-a method for simultaneously tackling two types of self-control problems by harnessing consumption complementarities. We describe a field experiment measuring the impact of bundling instantly gratifying but guilt-inducing "want" experiences (enjoying page-turner audiobooks) with valuable "should" behaviors providing delayed rewards (exercising). We explore whether such bundles increase should behaviors and whether people would pay to create these restrictive bundles. Participants were randomly assigned to a full treatment condition with gym-only access to tempting audio novels, an intermediate treatment involving encouragement to restrict audiobook enjoyment to the gym, or a control condition. Initially, full and intermediate treatment participants visited the gym 51% and 29% more frequently, respectively, than control participants, but treatment effects declined over time (particularly following Thanksgiving). After the study, 61% of participants opted to pay to have gym-only access to iPods containing tempting audiobooks, suggesting demand for this commitment device.


Common themes in nutrient acquisition by plant symbiotic microbes, described by the Gene Ontology.
A critical function for symbionts is the acquisition of nutrients from their host. Relationships between hosts and symbionts range from biotrophic mutualism to necrotrophic parasitism, with a corresponding range of structures to facilitate nutrient flow between host and symbiont. Here, we review common themes among the nutrient acquisition strategies of a range of plant symbiotic microorganisms, including mutualistic symbionts, biotrophic pathogens that feed from living tissue, necrotrophic pathogens that kill host tissue, and hemibiotrophic pathogens that switch from biotrophy to necrotrophy. We show how Gene Ontology (GO) terms developed by the Plant-Associated Microbe Gene Ontology (PAMGO) Consortium can be used for describing commonalities in nutrient acquisition among diverse plant symbionts. Where appropriate, parallels found among animal symbionts are also highlighted.


Subject level clustering using a negative binomial model for small transcriptomic studies.
Unsupervised clustering represents one of the most widely applied methods in analysis of high-throughput 'omics data. A variety of unsupervised model-based or parametric clustering methods and non-parametric clustering methods have been proposed for RNA-seq count data, most of which perform well for large samples, e.g. N ≥ 500. A common issue when analyzing limited samples of RNA-seq count data is that the data follows an over-dispersed distribution, and thus a Negative Binomial likelihood model is often used. Thus, we have developed a Negative Binomial model-based (NBMB) clustering approach for application to RNA-seq studies. We have developed a Negative Binomial Model-Based (NBMB) method to cluster samples using a stochastic version of the expectation-maximization algorithm. A simulation study involving various scenarios was completed to compare the performance of NBMB to Gaussian model-based or Gaussian mixture modeling (GMM). NBMB was also applied for the clustering of two RNA-seq studies; type 2 diabetes study (N = 96) and TCGA study of ovarian cancer (N = 295). Simulation results showed that NBMB outperforms GMM applied with different transformations in majority of scenarios with limited sample size. Additionally, we found that NBMB outperformed GMM for small clusters distance regardless of sample size. Increasing total number of genes with fixed proportion of differentially expressed genes does not change the outperformance of NBMB, but improves the overall performance of GMM. Analysis of type 2 diabetes and ovarian cancer tumor data with NBMB found good agreement with the reported disease subtypes and the gene expression patterns. This method is available in an R package on CRAN named NB.MClust. Use of Negative Binomial model based clustering is advisable when clustering over dispersed RNA-seq count data.


Lenalidomide in nonmetastatic biochemically relapsed prostate cancer: results of a phase I/II double-blinded, randomized study.
To evaluate the safety and activity of 6 months of treatment with lenalidomide at 5 or 25 mg/d in nonmetastatic biochemically relapsed prostate cancer. Sixty men with non-castrate, nonmetastatic, biochemically relapsed prostate cancer were stratified by prostate-specific antigen (PSA) doubling time, surgery/radiation therapy, prior androgen deprivation therapy (ADT), and randomized to lenalidomide 5 mg (n = 26) or 25 mg/d (n = 34) for 3 weeks repeated monthly for 6 months or until dose-limiting toxicity or disease progression. Toxicity was evaluated monthly, and PSAs and X-rays/scans every 6 months. Study size was determined to detect a progression rate of 40% at 6 months in either arm with 85% power (compared with a rate of 80% in the population receiving no treatment). Changes in PSA slopes were calculated using the regression of the log PSA for each patient before and during the initial 6 months and compared by t test. Baseline variables were balanced between arms. Grade 3/4 toxicity rates were 12% (n = 3) with 5 mg and 29% (n = 10) with 25 mg (P = 0.1), most commonly neutropenia (five patients, all on 25 mg). Two patients per arm had thromboembolic events. The change in PSA slope was greater with 25 mg versus 5 mg [-0.172 (-0.24 to -0.11) versus -0.033 (-0.11 to 0.04); P = 0.005]. With a mean follow-up of 31.4 months (range 14-44), five patients on 25 mg and one patient on 5 mg remain on the study. Lenalidomide has acceptable toxicity and is associated with long-term disease stabilization and PSA declines. Randomized studies evaluating conventional clinical disease end points in this patient population are planned.


Salmonella Combination Vaccines: Moving Beyond Typhoid.
There is now a robust pipeline of licensed and World Health Organization (WHO)-prequalified typhoid conjugate vaccines with a steady progression of national introductions. However, typhoid fever is responsible for less than half the total global burden of Salmonella disease, and even less among children aged <5 years. Invasive nontyphoidal Salmonella disease is the dominant clinical presentation of Salmonella in Africa, and over a quarter of enteric fever in Asia is due to paratyphoid A. In this article, we explore the case for combination Salmonella vaccines, review the current pipeline of these vaccines, and discuss key considerations for their development, including geographies of use, age of administration, and pathways to licensure. While a trivalent typhoid/nontyphoidal Salmonella vaccine is attractive for Africa, and a bivalent enteric fever vaccine for Asia, a quadrivalent vaccine covering the 4 main disease-causing serovars of Salmonella enterica would provide a single vaccine option for global Salmonella coverage.


Devices for insulin administration.
There is a significant need for revised, safe, and more effective insulin-delivery methods than subcutaneous injections in the treatment of both type I (insulin-dependent) and type II (non-insulin-dependent) diabetes. The aim of this review is to describe the rationale and methods for better use of injection and infusion devices for intensive insulin therapy and to describe results of animal and human research that will lead to an implantable artificial pancreas. Injection devices, e.g., jet injectors, insulin pens, and access ports, cannot be considered as a major breakthrough in the quest for improved control, although they may improve the patient's comfort. External pumps have benefits over multiple injections and conventional insulin therapy only in specific subgroups of patients, e.g., those with recurrent severe hypoglycemia, but only when used by experienced personnel. The external artificial pancreas (Biostator) is also to be used by experienced personnel for limited clinical and research applications, e.g., surgery of the diabetic patient. The development of an implantable version of the artificial pancreas is linked to progress in the field of reliable long-duration glucose sensors. Finally, programmable implantable insulin pumps, used as an open-loop delivery system, are the most promising alternative to intensive subcutaneous insulin strategies in the short term, although clear evidence of improved safety and efficacy remains to be documented.


Aerobic sludge granulation: a tale of two polysaccharides?
Aerobic sludge granules are suspended biofilms with the potential to reduce the cost and footprint of secondary wastewater treatment. Attempts to answer how and why they form leads to a consideration of the role of their extracellular polymeric substances (EPS) in determining their physical and microbiological properties. The exopolysaccharide components of this matrix, in particular, have received attention as putative structural, gel-forming agents. Two quite different exopolysaccharides have been proposed as the gel-forming constituents, with their gel properties clearly different from those of activated sludge EPS. This review aims to address the question of whether more than one gel-forming exopolysaccharide exist in granules. Based on the available structural data, it seems likely that they are different gel-forming polymers and their differences are not artifacts of the analytical methods used. Nonetheless, both proposed structural gel polymers are extracted and purified based on procedures selecting for anionic polar polysaccharides soluble at high pH, and both contain hexuronic acids. Granulation does not result from EPS synthesis by any single microbial population, nor from production of a single exopolysaccharide. Future studies using solvents suitable for recalcitrant polysaccharides are likely to reveal important structural roles for other polysaccharides. It is hoped that this article will serve as a guide for subsequent studies into understanding the roles of exopolysaccharides in aerobic granular sludge.


Receptor-mediated endocytosis of asialoglycoproteins by rat liver hepatocytes: biochemical characterization of the endosomal compartments.
The endocytic compartments of the asialoglycoprotein (ASGP) pathway in rat hepatocytes were studied using a combined morphological and biochemical approach in the isolated perfused liver. Use of electron microscopic tracers and a temperature-shift protocol to synchronize ligand entry confirmed the route of ASGP internalization observed in our previous in vivo studies (1) and established conditions under which we could label the contents of successive compartments in the pathway for subcellular fractionation studies. Three endosomal compartments were demonstrated in which ASGPs appear after they enter the cell via coated pits and vesicles but before they reach their site of degradation in lysosomes. These three compartments could be distinguished by their location within the hepatocyte, by their morphological appearance in situ, and by their density in sucrose gradients. The distributions of ASGP receptors, both accessible and latent (revealed by detergent permeabilization), were also examined and compared with that of ligand during subcellular fractionation. Most accessible ASGP receptors co-distributed with conventional plasma membrane markers. However, hepatocytes contain a substantial intracellular pool of latent ASGP binding sites that exceeds the number of cell surface receptors and whose presence is not dependent on ASGP exposure. The distribution of these latent ASGP receptors on sucrose gradients (detected either immunologically or by binding assays) was coincident with that of ligand sequestered within the early endosome compartments. In addition, both early endosomes and the membrane vesicles containing latent ASGP receptors had high cholesterol content, because both shifted markedly in density upon exposure to digitonin.


High-contrast single-particle tracking by selective focal plane illumination microscopy.
Wide-field single molecule microscopy is a versatile tool for analyzing dynamics and molecular interactions in biological systems. In extended three-dimensional systems, however, the method suffers from intrinsic out-of-focus fluorescence. We constructed a high-resolution selective plane illumination microscope (SPIM) to efficiently solve this problem. The instrument is an optical sectioning microscope featuring the high speed and high sensitivity of a video microscope. We present theoretical calculations and quantitative measurements of the illumination light sheet thickness yielding 1.7 microm (FWHM) at 543 nm, 2.0 microm at 633 nm, and a FWHM of the axial point spread function of 1.13 microm. A direct comparison of selective plane and epi-illumination of model samples with intrinsic background fluorescence illustrated the clear advantage of SPIM for such samples. Single fluorescent quantum dots in aqueous solution are readily visualized and tracked proving the suitability of our setup for the study of fast and dynamic processes in spatially extended biological specimens.


Evaluation of the root and canal systems of maxillary molars in Taiwanese patients: A cone beam computed tomography study.
This study evaluated variations in root canal configuration in the maxillary permanent molars of Taiwanese patients by analyzing patients' cone beam computed tomography (CBCT) images. Comparisons were made among these configurations and those previously reported. This information may serve as a basis for improving the success rate of endodontic treatment. The root canal systems of 114 Taiwanese patients with bilateral maxillary first or second molars were examined using CBCT images. The number of roots, canals per root, and additional mesiobuccal (MB) canals, as well as the canal configuration were enumerated and recorded. Of the 196 maxillary first molars examined, three (1.5%) had a single root, two (1.0%) had two roots, and 191 (97.5%) had three separate roots. Out of all first molar roots examined, 44% of mesiobuccal (MB) roots had a single canal and the remainder had a second MB (MB2) canal. Of the 212 maxillary second molars examined, 16 (7.1%) had a single root, 51 (24.2%) had two roots, 143 (67.8%) had three roots, and two (0.9%) had four separate roots. For the MB roots, 92.3% of three-rooted maxillary second molars had a single canal and the remainder had an MB2 canal. In all three-rooted maxillary first and second molars, each of the distal and palatal roots had one canal. The root canal configurations of the MB roots of maxillary molars were more varied than those of the distobuccal and palatal roots, and the root canal configurations of maxillary second molars were more varied than those of the first molars. These findings demonstrate CBCT as a useful clinical tool for endodontic diagnosis and treatment planning.


Does the multidisciplinary approach improve oncological outcomes in men undergoing surgical treatment for prostate cancer?
To determine whether oncological outcomes are improved in prostate cancer patients by using a multidisciplinary strategy as compared with a standard clinic paradigm, and whether time to treatment is delayed when using a multidisciplinary approach. We retrospectively analyzed patients who were evaluated and pursued radical prostatectomy as primary treatment, by the same surgeons, in the prostate cancer multidisciplinary clinic (n = 194) and standard urology clinic (n = 741) at Duke University Medical Center from 2005 to 2009. Comparisons of baseline characteristics were examined using rank sum and χ(2) -tests. Differences in time to radical prostatectomy and oncological outcomes were evaluated using multivariate linear and Cox regression, respectively. A greater proportion of high-risk patients (D'Amico criteria) were evaluated at the multidisciplinary clinic compared with the urology clinic (23.2% vs 15.6%, P = 0.014). Mean-adjusted time from biopsy to radical prostatectomy was shorter for multidisciplinary clinic patients (85.6 vs 96.8 days, P = 0.006). After a median follow up of 21 months, no significant difference was found between the multidisciplinary clinic and urology clinic in the risk of biochemical recurrence after radical prostatectomy, whether controlling for clinical (hazard ratio 0.71, P = 0.249) or pathological variables (hazard ratio 0.75, P = 0.349). Despite higher-risk disease, men evaluated using the multidisciplinary approach have similar oncological outcomes compared with men undergoing standard evaluation. Furthermore, time to radical prostatectomy is not delayed by the multidisciplinary management of these patients.


Effects of deep-bedded finishing system on market pig performance, composition and pork quality.
The purpose of this study was to compare effects of finishing environment on growth performance, pork quality and lipid composition of pork. Environments compared were standard confinement (CON) and deep-bedded semi-outdoor systems. The deep-bedded method employed in the current study was the use of hoop structures. Hoops are large, tent-like shelters with cornstalks or straw for bedding. Gilts ranging in weight from 59 to 71 kg were randomly assigned to treatments of Hoop (n = 50) and CON (n = 18) environments. Gilts were fed a two-phase dietary sequence, ad libitum for 45 days. Six gilts per treatment were selected for carcass composition and quality evaluation. The experiment was replicated a total of five times. Pigs raised in the Hoop environment gained significantly less and required significantly more feed for growth than pigs raised in the CON environment. Carcasses from CON-finished pigs were significantly fatter at the 10th rib, which lowered carcass percentage fat-free lean(FFL) and they also had greater loin marbling scores compared with carcasses from Hoop-finished pigs (P < 0.05). Significant replication effects were noted on beginning weight, live weight, carcass weight, percentage FFL, backfat, lipid content and adipose firmness. Carcasses from Hoop pigs had lower proportions of palmitic acid (P < 0.05), and higher proportions of oleic and linoleic acid (P < 0.05) in the inner layer of adipose tissue. The proportion of saturated fatty acid was lower, and that of mono- and poly-unsaturated fatty acid was higher in the inner layer of the adipose tissue of Hoop pigs. Variations in fatty acid composition and lipid deposition may have been caused by environmental temperature, since decreases in environmental temperature accompanied compositional variation of the adipose, leading to higher proportions of monounsaturated fatty acid and lower proportions of saturated and polyunsaturated fatty acid in adipose tissue, regardless of treatment. Volatile profile analysis revealed that adipose tissue of Hoop pigs had significantly higher amounts of 3-butanal and heptanal compared with CON pigs, which may be related to the amount of oleic and linoleic acids composing the adipose tissue. These data indicate finishing pigs in hoop structures allows for exposure to fluctuating temperatures, which may influence the growth of pigs, as well as fatty acid composition and firmness of pork products.


Romantic Attraction and Substance Use in 15-Year-Old Adolescents from Eight European Countries.
Sexual minority youth are at higher risk of substance use than heterosexual youth. However, most evidence in this area is from North America, and it is unclear whether the findings can be generalized to other cultures and countries. In this investigation, we used data from the 2014 Health Behaviour in School-aged Children (HBSC) study to compare substance use in same- and both-gender attracted 15-year-old adolescents from eight European countries (n = 14,545) to that of their peers who reported opposite-gender attraction or have not been romantically attracted to anyone. Both-gender attracted, and to a lesser extent, same-gender attracted adolescents were significantly more likely to smoke cigarettes, consume alcohol, get drunk and use cannabis, or be involved in multiple substance use in the last 30 days compared to their opposite-gender attracted peers. Those adolescents who have not been in love had significantly lower odds for substance use than all other youth. The pattern of results remained the same after adjusting for country, gender and family affluence. These findings are compatible with the minority stress and romantic stress theories. They suggest that sexual minority stigma (and love on its own) may contribute to higher substance use among adolescents in European countries.


Mitigating the effect of persistent postnatal depression on child outcomes through an intervention to treat depression and improve parenting: a randomised controlled trial.
Maternal postnatal depression occurs following 10-15% of births and is associated with a range of negative child outcomes. Risks to children are particularly increased when postnatal depression is persistent. We aimed to examine whether a parenting video-feedback therapy (VFT) intervention versus a control treatment of progressive muscle relaxation (PMR), both added to cognitive behavioural therapy (CBT) for persistent postnatal depression, would lead to improved child outcomes at age 2 years. In this two-arm, parallel-design, individually randomised controlled trial, we recruited a community sample of women aged 18 years or older living within 50 miles of Oxford, UK, between 4·5 and 9·0 months post partum. All participants met diagnostic criteria for current major depressive disorder that had persisted for at least 3 months and had infants at 35 or more weeks of gestation, with a birthweight of 2000 g or greater, and without serious neonatal complications. Through a centralised service, women were randomly assigned by use of a minimisation algorithm, to receive either VFT or PMR, balanced for child sex, temperament, age, socioeconomic status, and severity of depression. Both groups also received CBT for depression. Primary outcomes were child cognitive development, language development, behaviour problems, and attachment security at age 2 years. There were 11 home-based treatment sessions before child age 1 year, followed by two booster sessions in the second year. Assessors were masked to treatment group allocation. All analyses were done according to the intention-to-treat principle. This trial is registered with the ISRCTN registry, number ISRCTN07336477. Between March 18, 2011, and Dec 9, 2013, we randomly assigned 144 women, 72 to each group. Primary outcome data were available for 62-64 (86-89%) VFT and 67-68 (93-94%) PMR participants. There were no group differences in child outcome (cognitive development, adjusted difference -1·01 [95% CI -5·11 to 3·09], p=0·63; language development, 1·33 [-4·16 to 6·82], p=0·63; behaviour problems, -1·77 [-4·39 to 0·85], p=0·19; attachment security, 0·02 [-0·06 to 0·10], p=0·58), with both groups achieving scores similar to non-clinical norms on all outcomes. There were six serious adverse events: five in the VFT group (in two participants) and one in the PMR group. None was treatment-related. The effect of persistent postnatal depression on children is a major public health issue. For both treatment groups there was sustained remission from depression, and child development outcomes were in the normal range. The precise mechanisms accounting for the observed positive child outcomes cannot be ascertained from this study. Wellcome Trust.


Mathematical modelling of telomere length dynamics.
Telomeres are repetitive DNA sequences located at the ends of chromosomes. During cell division, an incomplete copy of each chromosome's DNA is made, causing telomeres to shorten on successive generations. When a threshold length is reached replication ceases and the cell becomes 'senescent'. In this paper, we consider populations of telomeres and, from discrete models, we derive partial differential equations which describe how the distribution of telomere lengths evolves over many generations. We initially consider a population of cells each containing just a single telomere. We use continuum models to compare the effects of various mechanisms of telomere shortening and rates of cell division during normal ageing. For example, the rate (or probability) of cell replication may be fixed or it may decrease as the telomeres shorten. Furthermore, the length of telomere lost on each replication may be constant, or may decrease as the telomeres shorten. Where possible, explicit solutions for the evolution of the distribution of telomere lengths are presented. In other cases, expressions for the mean of the distribution are derived. We extend the models to describe cell populations in which each cell contains a distinct subpopulation of chromosomes. As for the simpler models, constant telomere shortening leads to a linear reduction in telomere length over time, whereas length-dependent shortening results in initially rapid telomere length reduction, slowing at later times. Our analysis also reveals that constant telomere loss leads to a Gaussian (normal) distribution of telomere lengths, whereas length-dependent loss leads to a log-normal distribution. We show that stochastic models, which include a replication probability, also lead to telomere length distributions which are skewed.