Quantitative planar array screen of 1000 proteins uncovers novel urinary protein biomarkers of lupus nephritis.
The goal of these studies is to discover novel urinary biomarkers of lupus nephritis (LN). Urine from systemic lupus erythematosus (SLE) patients was interrogated for 1000 proteins using a novel, quantitative planar protein microarray. Hits were validated in an independent SLE cohort with inactive, active non-renal (ANR) and active renal (AR) patients, in a cohort with concurrent renal biopsies, and in a longitudinal cohort. Single-cell renal RNA sequencing data from LN kidneys were examined to deduce the cellular origin of each biomarker. Screening of 1000 proteins revealed 64 proteins to be significantly elevated in SLE urine, of which 17 were ELISA validated in independent cohorts. Urine Angptl4 (area under the curve (AUC)=0.96), L-selectin (AUC=0.86), TPP1 (AUC=0.84), transforming growth factor-β1 (TGFβ1) (AUC=0.78), thrombospondin-1 (AUC=0.73), FOLR2 (AUC=0.72), platelet-derived growth factor receptor-β (AUC=0.67) and PRX2 (AUC=0.65) distinguished AR from ANR SLE, outperforming anti-dsDNA, C3 and C4, in terms of specificity, sensitivity and positive predictive value. In multivariate regression analysis, urine Angptl4, L-selectin, TPP1 and TGFβ1 were highly associated with disease activity, even after correction for demographic variables. In SLE patients with serial follow-up, urine L-selectin (followed by urine Angptl4 and TGFβ1) were best at tracking concurrent or pending disease flares. Importantly, several proteins elevated in LN urine were also expressed within the kidneys in LN, either within resident renal cells or infiltrating immune cells, based on single-cell RNA sequencing analysis. Unbiased planar array screening of 1000 proteins has led to the discovery of urine Angptl4, L-selectin and TGFβ1 as potential biomarker candidates for tracking disease activity in LN.


Mass spectrometry to detect the site specificity of advanced glycation/lipoxidation end-product formation on protein: some challenges and solutions.
Formation of AGEs (advanced glycation end-products) and ALEs (advanced lipoxidation end-products) on proteins is associated with aging and various diseases of oxidative stress, notably diabetes and its complications. Modification of protein to AGE/ALEs is known to be site-directed and this has potential implications for protein functionality and design of AGE/ALE inhibitors. Determination of the site-specificity of modification is achieved most efficiently by MS. The present paper summarizes some of the challenges that need to be addressed when determining the site-specificity of AGE/ALE formation on protein by MS, using the protein RNase as an example. The following topics are discussed: formation and significance of AGE/ALEs, location of glycated peptides, enzymic digestion of glycated peptides and selection of mass spectrometric settings of analysis for glycated peptides.


Energy expenditure of bipedal walking is higher than that of quadrupedal walking in Japanese macaques.
The authors previously compared energetic costs of bipedal and quadrupedal walking in bipedally trained macaques used for traditional Japanese monkey performances (Nakatsukasa et al. 2004 Am. J. Phys. Anthropol. 124:248-256). These macaques used inverted pendulum mechanics during bipedal walking, which resulted in an efficient exchange of potential and kinetic energy. Nonetheless, energy expenditure during bipedal walking was significantly higher than that of quadrupedal walking. In Nakatsukasa et al. (2004 Am. J. Phys. Anthropol. 124:248-256), locomotor costs were measured before subjects reached a steady state due to technical limitations. The present investigation reports sequential changes of energy consumption during 15 min of walking in two trained macaques, using carbon dioxide production as a proxy of energy consumption, as in Nakatsukasa et al. (2004 Am. J. Phys. Anthropol. 124:248-256). Although a limited number of sessions were conducted, carbon dioxide production was consistently greater during bipedal walking, with the exception of some irregularity during the first minute. Carbon dioxide production gradually decreased after 1 min, and both subjects reached a steady state within 10 min. Energy expenditure during bipedalism relative to quadrupedalism differed between the two subjects. It was considerably higher (140% of the quadrupedal walking cost) in one subject who walked with more bent-knee, bent-hip gaits. This high cost strongly suggests that ordinary macaques, who adopt further bent-knee, bent-hip gaits, consume a far greater magnitude of energy during bipedal walking.


A cross-sectional study using freedom of information requests to evaluate variation in local authority commissioning of community pharmacy public health services in England.
This study aims to provide a national picture of the extent and nature of public health services commissioned by local authorities (LAs) from community pharmacies across England in financial year 2014/15. Cross-sectional survey of public health services commissioned in community pharmacies by LAs, gathered via freedom of information requests and documentary analysis. All 152 LAs in England. A total of 833 commissioned services were reported across England (range 3-10 per LA). Four services were commissioned by over 90% of LAs: emergency hormonal contraception (EHC), smoking cessation support, supervised consumption of methadone or other opiates and needle and syringe programmes (NSPs). The proportion of pharmacies commissioned to deliver these services varied considerably between LAs from <10% to 100%. This variation was not related to differences in relevant proxy measures of need. NHS Health Checks and alcohol screening and brief advice were commissioned by fewer LAs (32% and 15%, respectively), again with no relationship to relevant measures of need. A range of other services were commissioned less frequently, by fewer than 10% of LAs.Supervised consumption and NSPs were the most frequently used services, with over 4.4 million individual supervisions and over 1.4 million needle packs supplied. Pharmacies provided over 200 000 consultations for supply of EHC, over 30 000 supplies of free condoms and almost 16 000 chlamydia screening kits. More than 55 000 people registered to stop smoking in a community pharmacy, almost 30 000 were screened for alcohol use and over 26 000 NHS Health Checks were delivered. There is significant variation in commissioning and delivery of public health services in community pharmacies across England, which correlate poorly with potential benefit to local populations. Research to ascertain reasons for this variation is needed to ensure that future commissioning and delivery of these services matches local need.


Psychological effects of outdoor activity in type 2 diabetes: a review.
Previous research has shown individuals living with type 2 diabetes are more likely to experience depression, anxiety and reduced quality of life (QoL) in comparison to those without the disease. Physical activity has been highlighted as an important factor in improving physiological and psychological parameters in this population. Previous systematic reviews have found mixed results regarding the effects of exercise on psychological problems. Green exercise is an emerging research area with mounting evidence demonstrating positive effects on mental health. Objectives of the current review investigated whether conducting physical activity in outdoor environments had any effect on psychological problems in individuals with type 2 diabetes. Methods such as PubMed, Medline, PsycINFO, Scopus, Cochrane library, CINAHL, SPORTDiscus and Google Scholar databases were searched. The databases were searched for articles which used an outdoor exercise intervention and investigated psychological problems in individuals with type 2 diabetes. Results from the 1467 retrieved articles, 4 studies were included in the systematic review, consisting of 231 participants. All four studies investigated QoL, one study looked at depression and one investigated general wellbeing. Mixed results were found regarding QoL, no significant effect was found for depression. However, general wellbeing was improved. Due to the limited number of studies eligible for inclusion and the heterogeneity of outcome measures, it was difficult to draw firm conclusions. This systematic review acts as a starting point for future investigations into the use of green exercise in improving the mental health of individuals living with type 2 diabetes. Registration: PROSPERO: CRD42018088212.


Inadequate sleep as a contributor to type 2 diabetes in children and adolescents.
Lack of sleep is a modifiable risk factor for adverse health in humans. Short sleep duration and poor sleep quality are common in the pediatric population; the largest decline in sleep duration over the past decades has been seen in children and adolescents. The objective of the present narrative review was to provide for the first time an overview of the literature on sleep and its association with type 2 diabetes mellitus (T2D) biomarkers in children and adolescents. For this narrative review, 23 studies were retained (21 observational and 2 experimental studies). Notwithstanding the conflicting results found in these studies and despite being attenuated by adiposity level, maturity, sex and age, there is still some compelling evidence for an association between sleep duration (for both objective or subjective measurements of duration) and architecture with one or more T2D biomarkers in children and adolescents. The majority of the studies reviewed did focus on sleep duration and one or more T2D biomarkers in children and adolescents, but sleep architecture, more precisely the suppression of slow wave sleep and rapid eye movement sleep, has also been shown to be associated with insulin resistance. Only two studies looked at sleep quality, and the association between sleep quality and insulin resistance was not independent of level of adiposity. Future experimental studies will help to better understand the mechanisms linking insufficient sleep with T2D. Work also needs to be carried out on finding novel and effective strategies aimed at improving sleep hygiene and health outcomes of children and adolescents.


The Association of Breakfast Frequency and Cardiovascular Disease (CVD) Risk Factors among Adolescents in Malaysia.
Breakfast frequency is associated with cardiovascular disease (CVD) risk in Western populations, possibly via the types of food eaten or the timing of food consumption, but associations in Malaysian adolescents are unknown. While the timing of breakfast is similar, the type of food consumed at breakfast in Malaysia differs from Western diets, which allows novel insight into the mechanisms underlying breakfast-CVD risk associations. We investigated foods eaten for breakfast and associations between breakfast frequency and CVD risk factors in the Malaysian Health and Adolescents Longitudinal Research Team study (MyHeARTs). Breakfast (frequency of any food/drink reported as breakfast in 7-day diet history interviews) and CVD risk factors (body mass index (BMI), waist circumference, fasting blood glucose, triacylglycerol, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and systolic and diastolic blood pressure) were cross-sectionally associated using linear regression adjusting for potential confounders (n = 795, age 13 years). Twelve percent of adolescents never ate breakfast and 50% ate breakfast daily, containing mean (SD) 400 (±127) kilocalories. Commonly consumed breakfast foods were cereal-based dishes (primarily rice), confectionery (primarily sugar), hot/powdered drinks (primarily Milo), and high-fat milk (primarily sweetened condensed milk). After adjustment, each extra day of breakfast consumption per week was associated with a lower BMI (-0.34 kg/m2, 95% confidence interval (CI) -0.02, -0.66), and serum total (-0.07 mmol/L 95% CI -0.02, -0.13) and LDL (-0.07 mmol/L 95% CI -0.02, -0.12) cholesterol concentrations. Eating daily breakfast in Malaysia was associated with slightly lower BMI and total and LDL cholesterol concentrations among adolescents. Longitudinal studies and randomized trials could further establish causality.


GW151226: Observation of Gravitational Waves from a 22-Solar-Mass Binary Black Hole Coalescence.
We report the observation of a gravitational-wave signal produced by the coalescence of two stellar-mass black holes. The signal, GW151226, was observed by the twin detectors of the Laser Interferometer Gravitational-Wave Observatory (LIGO) on December 26, 2015 at 03:38:53 UTC. The signal was initially identified within 70 s by an online matched-filter search targeting binary coalescences. Subsequent off-line analyses recovered GW151226 with a network signal-to-noise ratio of 13 and a significance greater than 5σ. The signal persisted in the LIGO frequency band for approximately 1 s, increasing in frequency and amplitude over about 55 cycles from 35 to 450 Hz, and reached a peak gravitational strain of 3.4_{-0.9}^{+0.7}×10^{-22}. The inferred source-frame initial black hole masses are 14.2_{-3.7}^{+8.3}M_{⊙} and 7.5_{-2.3}^{+2.3}M_{⊙}, and the final black hole mass is 20.8_{-1.7}^{+6.1}M_{⊙}. We find that at least one of the component black holes has spin greater than 0.2. This source is located at a luminosity distance of 440_{-190}^{+180} Mpc corresponding to a redshift of 0.09_{-0.04}^{+0.03}. All uncertainties define a 90% credible interval. This second gravitational-wave observation provides improved constraints on stellar populations and on deviations from general relativity.


Therapeutic response monitoring after targeted therapy in an orthotopic rat model of hepatocellular carcinoma using contrast-enhanced ultrasound: Focusing on inter-scanner, and inter-operator reproducibility.
To assess therapeutic response monitoring after targeted therapy in an orthotopic rat model of hepatocellular carcinoma (HCC) using CEUS with focusing on inter-scanner and inter-operator reproducibility. For reproducibility, CEUS was performed using two different US scanners by two operators in sixteen rat models of HCC. Using perfusion analysis software (VueBox ®), eleven parameters were collected, and intra-class correlation coefficient (ICC) was used to analyze reproducibility. Then seventeen rat models of HCC were divided into treatment group (n = 8, 30 mg/kg/day sorafenib for five days) and control group (n = 9). CEUS was performed at baseline and 14 days after first treatment, and changes of perfusion parameters were analyzed. In treatment group, CEUS perfusion parameters showed a significant change. The peak enhancement (PE, 2.50 x103±1.68 x103 vs 5.55x102±4.65x102, p = 0.010) and wash-in and wash out AUC (WiWoAUC, 1.07x105±6.48 x104 vs 2.65x104±2.25x104, p = 0.009) had significantly decreased two weeks after treatment. On the contrary, control group did not show a significant change, including PE (1.15 x103±7.53x102 vs 9.43x102± 7.81 x102, p = 0.632) and WiWoAUC (5.09 x104±3.25x104 vs 5.92 x104±3.20x104, p = 0.646). For reproducibility, the various degrees of inter-scanner reproducibility were from poor to good (ICC: <0.01-0.63). However, inter-operator reproducibility of important perfusion parameters, including WiAUC, WoAUC, and WiWoAUC, ranged from fair to excellent (ICC: 0.59-0.93) in a different scanner. Our results suggest that CEUS is useful for assessment of the treatment response after targeted therapy and with fair to excellent inter-operator reproducibility.


Human immunodeficiency virus type 1 Tat activity in human neuronal cells: uptake and trans-activation.
Neurological dysfunction in AIDS occurs in the absence of productive infection of neurons, and may involve modulation of neuronal cell function by viral or cellular products released from surrounding infected cells. The human immunodeficiency virus type 1 (HIV-1) trans-activator protein Tat may be one such factor, as it can act as a neurotoxin, induces marked morphological changes in neurons and astrocytes in primary embryonic rodent brain cultures, and is released by certain HIV-1-infected cells. In addition, Tat can alter expression of cellular genes in several non-neuronal cell types. To explore the possibility that Tat may also mediate neuronal dysfunction in AIDS through non-lethal effects on neurons, we determined the trans-activating ability of Tat in human neuronal cells. We generated human neuronal cell lines stably expressing several HIV-1 tat genes, and also tested human neuronal cells exposed to extracellular recombinant Tat protein. Both endogenously expressed Tat as well as exogenous recombinant Tat protein up-regulated HIV-1 long terminal region (LTR)-driven gene expression by several hundred-fold. Only brief exposure to recombinant Tat was necessary and no toxic effects were seen at levels sufficient for trans-activation. Furthermore, Tat significantly enhanced virus expression in neuronal cells transfected with molecular clones of HIV-1. These results show that Tat is trans-activationally active in human neuronal cells, and can be taken up from the extracellular compartment by these cells in a biologically active form. Neurons represent an important potential target for Tat-mediated cellular dysfunction.


Evaluation of the effects of venlafaxine and pregabalin on the carbon dioxide inhalation models of Generalised Anxiety Disorder and panic.
Previous studies have shown that subjective and objective symptoms of anxiety induced by 7.5% CO(2) inhalation can be attenuated by anxiolytics such as lorazepam and, to a lesser extent, paroxetine. Venlafaxine and pregabalin, two other licensed treatments for Generalised Anxiety Disorder, were used to further investigate the 7.5% and 35% CO(2) models of anxiety in healthy volunteers. Fifty-four participants were randomised to receive either placebo, venlafaxine or pregabalin. Study treatments were dosed incrementally over a three week period, to reach daily doses of 150 mg venlafaxine and 200mg pregabalin by the CO(2) challenge test day. Participants inhaled air 7.5% CO(2) for 20 minutes (single-blind presentation), and a non-blinded single vital capacity of 35% CO(2). Subjective ratings were recorded before and after each inhalation. Both 7.5% and 35% CO(2) inhalations produced the expected effects of increased ratings of symptoms of panic and anxiety, with increased blood pressure and heart rate. No significant treatment effects were found, although there were trends towards a reduction in feeling tense and nervous by both drugs compared with placebo during the 7.5% CO(2) challenge, and a reduction in alertness generally in the venlafaxine group compared with the pregabalin group. In contrast with the clear anxiolytic effects of benzodiazepines reported in several previous CO(2) studies, these findings suggest that the anxiogenic effects of CO(2) challenges are not significantly influenced by these serotonergic and GABAergic anxiolytics. This may be due to a lack of sensitivity of the CO(2) challenges in healthy volunteers to these drug types.


The comparative risk of new-onset diabetes after prescription of drugs for cardiovascular risk prevention in primary care: a national cohort study.
Recent studies suggest that statins increase the risk of subsequent diabetes with a clear dose response effect. However, patients prescribed statins have a higher background risk of diabetes. This national cohort study aims to provide an estimate of the comparative risks for subsequent development of new-onset diabetes in adults prescribed statins and in those with an already higher background risk on cardiovascular risk-modifying drugs and a control drug. Longitudinal cohort study. Use of routinely collected data from a complete national primary care electronic prescription database in New Zealand. 32 086 patients aged between 40 and 60 years in 2005 were eligible and assigned to four non-overlapping groups receiving their first prescription for: (1) diclofenac (healthy population) n=7140; (2) antihypertensives thought likely to induce diabetes (thiazides and β-blockers) n=5769; (3) antihypertensives thought less likely to induce diabetes (ACE inhibitors, angiotensin II receptor blockers, calcium channel blocker) n=6565 and (4) statins n=12 612. Numbers of first metformin prescriptions were compared between these groups from 2006 to 2011. Patients prescribed statins have the highest risk of receiving a subsequent metformin prescription (HR 3.31; 95% CI 2.56 to 4.30; p<0.01), followed by patients prescribed antihypertensives thought less likely to induce diabetes (HR 2.32; 95% CI 1.74 to 3.09; p<0.01) and patients prescribed antihypertensives thought more likely to induce diabetes (HR 1.59; 95% CI 1.15 to 2.20; p<0.01) in the subsequent 6 years of follow-up, when compared to diclofenac. These findings further support the link between statin use and new-onset diabetes and suggest that the understanding of diabetes risk associated with different antihypertensive drug classes may bear practice modification. This provides important information for future research, and for prescribers and patients when considering the risks and benefits of different types of cardiovascular risk-modifying drugs.


The Lats2 tumor suppressor augments p53-mediated apoptosis by promoting the nuclear proapoptotic function of ASPP1.
Apoptosis is an important mechanism to eliminate potentially tumorigenic cells. The tumor suppressor p53 plays a pivotal role in this process. Many tumors harbor mutant p53, but others evade its tumor-suppressive effects by altering the expression of proteins that regulate the p53 pathway. ASPP1 (apoptosis-stimulating protein of p53-1) is a key mediator of the nuclear p53 apoptotic response. Under basal conditions, ASPP1 is cytoplasmic. We report that, in response to oncogenic stress, the tumor suppressor Lats2 (large tumor suppressor 2) phosphorylates ASPP1 and drives its translocation into the nucleus. Together, Lats2 and ASPP1 shunt p53 to proapoptotic promoters and promote the death of polyploid cells. These effects are overridden by the Yap1 (Yes-associated protein 1) oncoprotein, which disrupts Lats2-ASPP1 binding and antagonizes the tumor-suppressing function of the Lats2/ASPP1/p53 axis.


Whole genome sequence analysis of pulmonary function and COPD in 19,996 multi-ethnic participants.
Chronic obstructive pulmonary disease (COPD), diagnosed by reduced lung function, is a leading cause of morbidity and mortality. We performed whole genome sequence (WGS) analysis of lung function and COPD in a multi-ethnic sample of 11,497 participants from population- and family-based studies, and 8499 individuals from COPD-enriched studies in the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program. We identify at genome-wide significance 10 known GWAS loci and 22 distinct, previously unreported loci, including two common variant signals from stratified analysis of African Americans. Four novel common variants within the regions of PIAS1, RGN (two variants) and FTO show evidence of replication in the UK Biobank (European ancestry n ~ 320,000), while colocalization analyses leveraging multi-omic data from GTEx and TOPMed identify potential molecular mechanisms underlying four of the 22 novel loci. Our study demonstrates the value of performing WGS analyses and multi-omic follow-up in cohorts of diverse ancestry.


Exploring local public health work in the context of novel translation technologies.
Limited English proficiency (LEP) leads to disparities in health and health care. Although the LEP population in the US is growing and federal regulations require availability of translated health promotion materials, availability of these materials is lacking, partially due to the costs and time required to produce high quality translations. We conducted semi-structured interviews with employees involved in the translation process at three local health districts (LHDs) in Washington State to explore the potential for novel technologies to assist in the creation of multilingual health promotion materials. We generated a translation workflow diagram and analyzed key themes that affected the translation process. We also explored employees' opinions regarding electronic document exchange and machine translation technology (MT). Overall, public health employees were open to the incorporation of information technologies to speed the translation and dissemination of multilingual public health materials if quality could be assured.


Alcohol, appetite and energy balance: is alcohol intake a risk factor for obesity?
The increased recognition that the worldwide increase in incidence of obesity is due to a positive energy balance has lead to a focus on lifestyle choices that may contribute to excess energy intake, including the widespread belief that alcohol intake is a significant risk factor for development of obesity. This brief review examines this issue by contrasting short-term laboratory-based studies of the effects of alcohol on appetite and energy balance and longer-term epidemiological data exploring the relationship between alcohol intake and body weight. Current research clearly shows that energy consumed as alcohol is additive to that from other dietary sources, leading to short-term passive over-consumption of energy when alcohol is consumed. Indeed, alcohol consumed before or with meals tends to increase food intake, probably through enhancing the short-term rewarding effects of food. However, while these data might suggest that alcohol is a risk factor for obesity, epidemiological data suggests that moderate alcohol intake may protect against obesity, particularly in women. In contrast, higher intakes of alcohol in the absence of alcohol dependence may increase the risk of obesity, as may binge-drinking, however these effects may be secondary to personality and habitual beverage preferences.


Symbolic discriminant analysis of microarray data in autoimmune disease.
New laboratory technologies such as DNA microarrays have made it possible to measure the expression levels of thousands of genes simultaneously in a particular cell or tissue. The challenge for genetic epidemiologists will be to develop statistical and computational methods that are able to identify subsets of gene expression variables that classify and predict clinical endpoints. Linear discriminant analysis is a popular multivariate statistical approach for classification of observations into groups. This is because the theory is well described and the method is easy to implement and interpret. However, an important limitation is that linear discriminant functions need to be prespecified. To address this limitation and the limitation of linearity, we have developed symbolic discriminant analysis (SDA) for the automatic selection of gene expression variables and discriminant functions that can take any form. In the present study, we demonstrate that SDA is capable of identifying combinations of gene expression variables that are able to classify and predict autoimmune diseases.


Trophodynamics and mercury bioaccumulation in reef and open-ocean fishes from The Bahamas with a focus on two teleost predators.
Identifying prey resource pools supporting fish biomass can elucidate trophic pathways of pollutant bioaccumulation. We used multiple chemical tracers (carbon [δ13C] and nitrogen [δ15N] stable isotopes and total mercury [THg]) to identify trophic pathways and measure contaminant loading in upper trophic level fishes residing at a reef and open-ocean interface near Eleuthera in the Exuma Sound, The Bahamas. We focused predominantly on the trophic pathways of mercury bioaccumulation in dolphinfish Coryphaena hippurus and wahoo Acanthocybium solandri, 2 commonly consumed pelagic sportfish in the region. Despite residing within close proximity to productive and extensive coral reefs, both dolphinfish and wahoo relied almost exclusively on open-ocean prey over both short and long temporal durations. A larger isotopic niche of dolphinfish suggested a broader diet and some potential prey differentiation between the 2 species. THg concentrations in dolphinfish (0.2 ± 0.1 ppm) and wahoo (0.3 ± 0.3 ppm) were mostly below recommended guidelines for humans (US Environmental Protection Agency (EPA) = 0.3 ppm, US Food and Drug Administration (FDA)= 1.0 ppm) and were within ranges previously reported for these species. However, high THg concentrations were observed in muscle and liver tissue of commonly consumed reef-associated fishes, identifying a previously unrecognized route of potentially toxic Hg exposure for human consumers on Eleuthera and neighboring islands.


Streptococcus pneumoniae Binds to Host Lactate Dehydrogenase via PspA and PspC To Enhance Virulence.
Pneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC, also called CbpA) are major virulence factors of Streptococcus pneumoniae (Spn). These surface-exposed choline-binding proteins (CBPs) function independently to inhibit opsonization, neutralize antimicrobial factors, or serve as adhesins. PspA and PspC both carry a proline-rich domain (PRD) whose role, other than serving as a flexible connector between the N-terminal and C-terminal domains, was up to this point unknown. Herein, we demonstrate that PspA binds to lactate dehydrogenase (LDH) released from dying host cells during infection. Using recombinant versions of PspA and isogenic mutants lacking PspA or specific domains of PspA, this property was mapped to a conserved 22-amino-acid nonproline block (NPB) found within the PRD of most PspAs and PspCs. The NPB of PspA had specific affinity for LDH-A, which converts pyruvate to lactate. In a mouse model of pneumonia, preincubation of Spn carrying NPB-bearing PspA with LDH-A resulted in increased bacterial titers in the lungs. In contrast, incubation of Spn carrying a version of PspA lacking the NPB with LDH-A or incubation of wild-type Spn with enzymatically inactive LDH-A did not enhance virulence. Preincubation of NPB-bearing Spn with lactate alone enhanced virulence in a pneumonia model, indicating exogenous lactate production by Spn-bound LDH-A had an important role in pneumococcal pathogenesis. Our observations show that lung LDH, released during the infection, is an important binding target for Spn via PspA/PspC and that pneumococci utilize LDH-A derived lactate for their benefit in vivoIMPORTANCEStreptococcus pneumoniae (Spn) is the leading cause of community-acquired pneumonia. PspA and PspC are among its most important virulence factors, and these surface proteins carry the proline-rich domain (PRD), whose role was unknown until now. Herein, we show that a conserved 22-amino-acid nonproline block (NPB) found within most versions of the PRD binds to host-derived lactate dehydrogenase A (LDH-A), a metabolic enzyme which converts pyruvate to lactate. PspA-mediated binding of LDH-A increased Spn titers in the lungs and this required LDH-A enzymatic activity. Enhanced virulence was also observed when Spn was preincubated with lactate, suggesting LDH-A-derived lactate is a vital food source. Our findings define a role for the NPB of the PRD and show that Spn co-opts host enzymes for its benefit. They advance our understanding of pneumococcal pathogenesis and have key implications on the susceptibility of individuals with preexisting airway damage that results in LDH-A release.


Coordination of WNT signaling and ciliogenesis during odontogenesis by piezo type mechanosensitive ion channel component 1.
Signal transmission from the mechanical forces to the various intracellular activities is a fundamental process during tissue development. Despite their critical role, the mechanism of mechanical forces in the biological process is poorly understood. In this study, we demonstrated that in the response to hydrostatic pressure (HP), the piezo type mechanosensitive ion channel component 1 (PIEZO1) is a primary mechanosensing receptor for odontoblast differentiation through coordination of the WNT expression and ciliogenesis. In stem cells from human exfoliated deciduous teeth (SHED), HP significantly promoted calcium deposition as well as the expression of odontogenic marker genes, PANX3 and DSPP, and WNT related-genes including WNT5b and WNT16, whereas HP inhibited cell proliferation and enhanced primary cilia expression. WNT signaling inhibitor XAV939 and primary cilia inhibitor chloral hydrate blocked the HP-induced calcium deposition. The PIEZO1 activator Yoda1 inhibited cell proliferation but induced ciliogenesis and WNT16 expression. Interestingly, HP and Yoda1 promoted nuclear translocation of RUNX2, whereas siRNA-mediated silencing of PIEZO1 decreased HP-induced nuclear translocation of RUNX2. Taken together, these results suggest that PIEZO1 functions as a mechanotransducer that connects HP signal to the intracellular signalings during odontoblast differentiation.


Self-reported adverse events in health care that cause harm: a population-based survey.
To identify the incidence of self-reported harmful adverse events in the health care of community-dwelling adults, and to examine attitudes about safety in the health system. Cross-sectional, population-based survey. Analysis of data from 3522 adults participating in Stage 2 of the North West Adelaide Health Study, who were surveyed in 2004 and 2005. Self-reported adverse events causing harm in the past year; attitudes to safety in health care, including adequacy of current measures for preventing adverse events, and the effect of groups like patients themselves, doctors and governments on patient safety. The annual incidence of self-reported harmful adverse events was 4.2%. The main types were medication error (45.5%) and misdiagnosis or wrong treatment (25.6%). Multiple logistic regression showed that self-reported harmful adverse events were more likely in people who had been hospitalised in the past 12 months (odds ratio [OR], 2.5; 95% CI, 1.9-3.4), those who had low annual income (< $12 000), those who completed higher education to the level of Bachelor degree or higher (OR, 3.0; 95% CI, 1.0-9.4), and those who had some level of dissatisfaction with their recent health care; and less likely in those more risk-averse rather than those with a tendency to risk-taking behaviour (OR, 0.6; 95% CI, 0.4-0.9). People were more likely to believe that individual health care professionals had a positive effect on safety than professional groups or government, and that more resources were the key to improving the safety of health care. We found an incidence of self-reported harmful adverse events that was significantly lower than that found by a 2002 Australian survey (4.2% v 6.5%; P = 0.009). Better communication to help patients acquire more realistic risk perception may help reduce harm. Better communication could also increase public advocacy for systems improvement in safety to counter persisting community beliefs that individual action alone can redress the situation.


Evaluation of the capacity to appoint a healthcare proxy.
The appointment of a healthcare proxy is the most common way through which patients appoint a surrogate decision maker in anticipation of a future time in which they may lack the ability to make medical decisions themselves. In some situations, when a patient has not previously appointed a surrogate decision maker through an advance directive, the healthcare team may ask whether the patient, although lacking the capacity to make a healthcare decision, might still have the capacity to appoint a healthcare proxy. In this article the authors summarize the existing, albeit limited, legal and empirical basis for this capacity and propose a model for assessing capacity to appoint a healthcare proxy that incorporates clinical factors in the context of the risks and benefits specific to surrogate appointment under the law. In particular, it is important to weigh patients' understanding and choice within the context of the risks and benefits of the medical and interpersonal factors. Questions to guide capacity assessment are provided for clinical use and refinement through future research.


High-resolution MR venography at 3.0 Tesla.
The aim of this study was to investigate the visualization of small venous vessels in the normal human brain at a field strength of 3 Tesla. T2*-weighted, three-dimensional gradient-echo images were acquired by exploiting the magnetic susceptibility difference between oxygenated and deoxygenated hemoglobin in the vasculature and microvasculature. The spatial resolution was 0.5 x 0.5 x 1 mm3, and sequence parameters were varied to obtain good vessel delineation. Improved visibility of venous vessels was obtained by creating phase mask images from the magnetic resonance phase images and multiplying these by the magnitude images. Venograms were created by performing a minimum intensity projection over targeted volumes. Highly detailed visualization of venous structures deep in the brain and in the superficial cortical areas were obtained without administration of an exogenous contrast agent; compared with similar studies performed at 1.5 T, the echo time could be reduced from typically 40-50 ms to 17-28 ms. Imaging at high-field strength offers the possibility of improved resolution and the delineation of smaller vessels compared with lower field strengths.


Social inheritance can explain the structure of animal social networks.
The social network structure of animal populations has major implications for survival, reproductive success, sexual selection and pathogen transmission of individuals. But as of yet, no general theory of social network structure exists that can explain the diversity of social networks observed in nature, and serve as a null model for detecting species and population-specific factors. Here we propose a simple and generally applicable model of social network structure. We consider the emergence of network structure as a result of social inheritance, in which newborns are likely to bond with maternal contacts, and via forming bonds randomly. We compare model output with data from several species, showing that it can generate networks with properties such as those observed in real social systems. Our model demonstrates that important observed properties of social networks, including heritability of network position or assortative associations, can be understood as consequences of social inheritance.


Association Between Behavioral and Learning Outcomes and Single Exposures to Procedures Requiring General Anesthesia Before Age 3: Secondary Analysis of Data From Olmsted County, MN.
Two prior population-based (children born in Olmsted County, MN), retrospective cohort studies both found that multiple exposures to anesthesia before age 3 were associated with a significant increase in the frequency of attention-deficit hyperactivity disorder (ADHD) and learning disabilities (LD) later in life. The primary purpose of this secondary analysis of these data was to test the hypothesis that a single exposure to anesthesia before age 3 was associated with an increased risk of ADHD. We also examined the association of single exposures with LD and the need for individualized educational plans as secondary outcomes. This analysis includes 5339 children who were unexposed to general anesthesia before age 3 (4876 born from 1976 to 1982 and 463 born from 1996 to 2000), and 1054 children who had a single exposure to anesthesia before age 3 (481 born from 1976 to 1982 and 573 born from 1996 to 2000). The primary outcome of interest was ADHD. Secondary outcomes included LD (reading, mathematics, and written language) and the need for individualized educational programs (speech/language and emotion/behavior). To compare the incidence of each outcome between those who were unexposed and singly exposed to anesthesia before the age of 3 years, an inverse probability of treatment weighted proportional hazards model was used. For children not exposed to anesthesia, the estimated cumulative frequency (95% confidence interval [CI]) of ADHD at age 18 was 7.3% (95% CI, 6.5-8.1) and 13.0% (95% CI, 10.1-16.8) for the 1976-1982 and 1996-2000 cohorts, respectively. For children exposed to a single anesthetic before age 3, the cumulative frequency of ADHD was 8.1% (95% CI, 5.3-12.4) and 17.6% (95% CI, 14.0-21.9) for the 1976-1982 and 1996-2000 cohorts, respectively. In weighted analyses, single exposures were not significantly associated with an increased frequency of ADHD (hazard ratio [HR], 1.21; 95% CI, 0.91-1.60; P = .184). Single exposures were also not associated with an increased frequency of any LD (HR, 0.98; 95% CI, 0.78-1.23), or the need for individualized education plans. This analysis did not find evidence that single exposures to procedures requiring general anesthesia, before age 3, are associated with an increased risk of developing ADHD, LD, or the need for individualized educational plans in later life.


Intrastriatal transplantation of adult human neural crest-derived stem cells improves functional outcome in parkinsonian rats.
Parkinson's disease (PD) is considered the second most frequent and one of the most severe neurodegenerative diseases, with dysfunctions of the motor system and with nonmotor symptoms such as depression and dementia. Compensation for the progressive loss of dopaminergic (DA) neurons during PD using current pharmacological treatment strategies is limited and remains challenging. Pluripotent stem cell-based regenerative medicine may offer a promising therapeutic alternative, although the medical application of human embryonic tissue and pluripotent stem cells is still a matter of ethical and practical debate. Addressing these challenges, the present study investigated the potential of adult human neural crest-derived stem cells derived from the inferior turbinate (ITSCs) transplanted into a parkinsonian rat model. Emphasizing their capability to give rise to nervous tissue, ITSCs isolated from the adult human nose efficiently differentiated into functional mature neurons in vitro. Additional successful dopaminergic differentiation of ITSCs was subsequently followed by their transplantation into a unilaterally lesioned 6-hydroxydopamine rat PD model. Transplantation of predifferentiated or undifferentiated ITSCs led to robust restoration of rotational behavior, accompanied by significant recovery of DA neurons within the substantia nigra. ITSCs were further shown to migrate extensively in loose streams primarily toward the posterior direction as far as to the midbrain region, at which point they were able to differentiate into DA neurons within the locus ceruleus. We demonstrate, for the first time, that adult human ITSCs are capable of functionally recovering a PD rat model.


Questionnaire study of the dietary habits of breast cancer survivors and their relationship to quality of life (KROG 14-09).
We evaluated the dietary habits of breast cancer survivors and investigated the relationship with quality of life (QoL), with 1,156 survivors recruited from 17 institutions. We used the Questionnaire Survey of Dietary Habits of Korean Adults (Q-DH-KOR) comprising 25 questions. The following indices were derived as follows: (1) quality of healthy dietary habits (Q-HD)-eight questions on number of meals, regularity, quantity, duration, skipping breakfast, dinner with companion(s), overeating and late-night snacks; (2) habits of nutritional balance (H-NB)-questions on consuming five food categories (grains, fruits, proteins, vegetables and dairy products); and (3) habits of unhealthy foods (H-UF)-questions on consuming three food categories (fatty, instant and fast foods). The times and regularity of meals, frequency of skipping breakfast, dinner with companion(s) and overeating were better in groups with high symptomatic and functional QoL. Symptomatic QoL positively affected Q-HD and H-NB (p < 0.001 and p = 0.024 respectively) and negatively affected H-UF (p = 0.02). Breast cancer survivors more frequently ate from the fruit, protein and vegetable categories than did the control group, with lower H-UF and higher Q-HD values (p < 0.001 and p < 0.001 respectively). Our findings supported the relationship between QoL and dietary habit and showed healthier dietary habits of breast cancer survivors than controls.


Vancomycin-Induced Kidney Injury: Animal Models of Toxicodynamics, Mechanisms of Injury, Human Translation, and Potential Strategies for Prevention.
Vancomycin is a recommended therapy in multiple national guidelines. Despite the common use, there is a poor understanding of the mechanistic drivers and potential modifiers of vancomycin-mediated kidney injury. In this review, historic and contemporary rates of vancomycin-induced kidney injury (VIKI) are described, and toxicodynamic models and mechanisms of toxicity from preclinical studies are reviewed. Aside from known clinical covariates that worsen VIKI, preclinical models have demonstrated that various factors impact VIKI, including dose, route of administration, and thresholds for pharmacokinetic parameters. The degree of acute kidney injury (AKI) is greatest with the intravenous route and higher doses that produce larger maximal concentrations and areas under the concentration curve. Troughs (i.e., minimum concentrations) have less of an impact. Mechanistically, preclinical studies have identified that VIKI is a result of drug accumulation in proximal tubule cells, which triggers cellular oxidative stress and apoptosis. Yet, there are several gaps in the knowledge that may represent viable targets to make vancomycin therapy less toxic. Potential strategies include prolonging infusions and lowering maximal concentrations, administration of antioxidants, administering agents that decrease cellular accumulation, and reformulating vancomycin to alter the renal clearance mechanism. Based on preclinical models and mechanisms of toxicity, we propose potential strategies to lessen VIKI.


Post-infection immunocomplex glomerulonephritis and Legionnaires' disease in a patient with adult Still's disease during treatment with interleukin 1 receptor antagonist anakinra: a case report.
Legionellosis is a systemic disease that primarily affects the lungs. However, dysfunction in many organ systems, including the kidneys, has also been described. There are only a few reported cases of renal dysfunction in patients with legionellosis. A 27-year-old Caucasian woman with known adult Still's disease was admitted to our hospital for community-acquired pneumonia, due to Legionella infection, with acute renal failure. Although her respiratory symptoms responded well to antibiotic treatment, her renal function worsened, with severe proteinuria and edema. A renal biopsy showed extracapillary and endocapillary proliferative glomerulonephritis with accompanying chronic and acute interstitial nephritis. This was consistent with a post-infection immunocomplex glomerulonephritis. After initiation of steroid therapy, her renal function improved. Additionally, therapy with diuretics and an angiotensin-converting enzyme inhibitor was initiated because of persistent proteinuria. Under this treatment regimen, her severe edema and proteinuria disappeared. To the best of our knowledge, there is only a handful of reported cases of post-infection glomerulonephritis with a nephrotic syndrome in a patient with legionellosis. Our findings suggest that, in patients with Legionnaires' disease with renal failure, post-infection immunocomplex glomerulonephritis should be considered and steroid therapy may be an effective modality to treat the renal complication.


Thiazolidinediones inhibit the expression of beta3-adrenergic receptors at a transcriptional level.
The effect of the thiazolidinediones (TZDs) darglitazone and troglitazone on beta3-adrenergic receptor (AR) expression was studied in cultured cell lines representing several tissues. After 24 h of exposing HIB-1B brown adipocytes to 30 micromol/l darglitazone or 20 micromol/l troglitazone, beta3-AR mRNA levels were reduced by 75%. This effect was significant within 1 h of exposure to a maximal dose of these drugs, with the full effect obtained within 10 h. The darglitazone ID50 was approximately 10 nmol/l, similar to the Kd of TZDs binding to peroxisome proliferator-activated receptor-gamma (PPAR-gamma). These drugs also decreased beta3-AR mRNA in 3T3-F442A white adipocytes, but not in SK-N-MC cells, which lack PPAR-gamma2. A luciferase reporter gene containing 1.4 kb of 5' flanking sequence of the mouse beta3-AR was transiently transfected, with or without PPAR-gamma2, in SK-N-MC cells. The vigorous expression of luciferase driven by the beta3-AR gene sequence was inhibited by TZDs in a PPAR-gamma2-dependent manner. The half-lives of gamma3-AR precursor RNA and mRNA were short, approximately 40 and approximately 100 min, respectively, and remained unaffected by TZD treatment. Exposure of HIB-1B cells to 30 micromol/l darglitazone was associated with reduced beta3-AR mRNA levels, as well as decreased response of uncoupling protein 1 to norepinephrine + propranolol (a beta1 beta2-AR antagonist) or the specific beta3-AR agonist CL 316, 243. Both the beta3-AR mRNA level and response to these stimuli fully recovered by 24 h of removing the drug, indicating that the beta3-AR protein and its coupling to adenylyl cyclase rapidly followed the changes in mRNA. Thus, TZDs can rapidly reduce beta3-AR expression at the transcriptional level, acting through PPAR-gamma2. The rapid turnover and responses of beta3-AR to perturbations, along with numerous other factors reported to regulate its expression, suggest a tight control of beta3-AR and function. Lastly, leptin being the only other known gene suppressed by TZDs, the present studies support a concerted lipogenic effect of these drugs.


Predictors and Causes of Long-Term Mortality in Elderly Patients with Acute Venous Thromboembolism: A Prospective Cohort Study.
Long-term predictors and causes of death are understudied in elderly patients with acute venous thromboembolism. We prospectively followed up 991 patients aged ≥65 years with acute venous thromboembolism in a multicenter Swiss cohort study. The primary outcome was overall mortality. We explored the association between patient baseline characteristics and mortality, adjusting for other baseline variables and periods of anticoagulation as a time-varying covariate. Causes of death over time were adjudicated by a blinded, independent committee. The median age was 75 years. During a median follow-up period of 30 months, 206 patients (21%) died. Independent predictors of overall mortality were age (hazard ratio [HR], 1.32; 95% confidence interval [CI], 1.05-1.65, per decade), active cancer (HR, 5.80; 95% CI, 4.22-7.97), systolic blood pressure <100 mm Hg (HR, 2.77; 95% CI, 1.56-4.92), diabetes mellitus (HR, 1.50; 95% CI, 1.02-2.22), low physical activity level (HR, 1.92; 95% CI, 1.38-2.66), polypharmacy (HR, 1.41; 95% CI, 1.01-1.96), anemia (HR, 1.48; 95% CI, 1.07-2.05), high-sensitivity C-reactive protein >40 mg/L (HR, 1.88; 95% CI, 1.36-2.60), ultra-sensitive troponin >14 pg/mL (HR, 1.54; 95% CI, 1.06-2.25), and D-dimer >3000 ng/mL (HR, 1.45; 95% CI, 1.04-2.01). Cancer (34%), pulmonary embolism (18%), infection (17%), and bleeding (6%) were the most common causes of death. Elderly patients with acute venous thromboembolism have a substantial long-term mortality, and several factors, including polypharmacy and a low physical activity level, are associated with long-term mortality. Cancer, pulmonary embolism, infections, and bleeding are the most common causes of death in the elderly with venous thromboembolism.


WE-G-217BCD-09: Calibration of a DRR Algorithm.
To calibrate an algorithm for digitally reconstructed radiographs (DRRs) such that synthetic projections cast through an object in a fan-beam CT (FBCT) optimally match cone-beam CT projections of the same object. Our DRR algorithm models the transmission of primary photons through an object with a ray-tracing algorithm that samples the CT at small steps along each source-detector pixel path. Sampled CT values are converted to linear attenuation coefficients (LACs), scaled by the step-size, and added to a running sum for each detector pixel. We made attenuation measurements of materials of known CT number to calibrate a CT to LAC conversion function for our FBCT and CBCT systems. DRRs are post- processed to reproduce the tube output, source fluence distribution, and detector response of the CBCT system. In addition, they are modified to account for scatter, beam hardening, and detector veiling glare. Finally, we determined CBCT geometry parameters using a specially designed phantom. We analyzed the quality of our DRR algorithm by directly comparing synthetic DRRs cast through the FBCT of a Catphan® phantom with actual CBCT projections of the phantom. Line plots comparing the intensity profiles of the DRRs and CBCT projections show that uncorrected DRRs do not align perfectly with the projections. However, when scatter, beam hardening, and veiling glare are modeled there is much closer agreement. Simulations with different geometry phantom shapes and sizes recommend a cylindrical phantom 150 mm in diameter. Computational times under one second per DRR are achieved using a GPU for a simulated 1024×768 pixel detector with a 512×512×281 mm CT volume and 0.5 mm ray step-size. Our DRR algorithm is able to closely reproduce actual CBCT projections taken through a test object. It is optimized specifically for the FBCT and CBCT systems in our department. Funded in part by NCI grants R01CA123299 and P01CA116602. The authors report no conflicts of interest.


Emissions of organic compounds and trace metals in fine particulate matter from motor vehicles: a tunnel study in Houston, Texas.
Fine particulate matter (PM) samples collected in a highway tunnel in Houston, TX, were analyzed to quantify the concentrations of 14 n-alkanes, 12 polycyclic aromatic hydrocarbons, and nine petroleum biomarkers, as well as 21 metals, with the ultimate aim of identifying appropriate tracers for diesel engines. First, an exploratory multivariate dimensionality reduction technique called principal component analysis (PCA) was employed to identify all potential candidates for tracers. Next, emission indices were calculated to interpret PCA results physically. Emission indices of n-heneicosane, n-docosane, n-tricosane, n-tetracosane, n-pentacosane, fluoranthene, and pyrene were correlated highly and increased strongly with percentage carbon present in the tunnel emanating from diesel vehicles. This suggests that these organic compounds are useful molecular markers to separate emissions from diesel and gasoline engines. Additionally, the results are the first quantification of the metal composition of PM with aerodynamic diameters smaller than 2.5 microm (PM2.5) emissions from mobile sources in Houston. PCA of trace metal concentrations followed by emission index calculations revealed that barium in fine airborne particles can be linked quantitatively to diesel engine emissions, demonstrating its role as an elemental tracer for heavy-duty trucks.


Trephine Technique for Iliac Crest Bone Graft Harvest: Long-term Results.
Report of surgical technique and long-term outcome; review of prospectively gathered data and follow-up survey of patient cohort. The aim of this study was to review surgical technique for a trephine graft harvest technique and assess perioperative complications/pain, long-term outcome, and patient satisfaction after anterior iliac crest bone graft (ICBG) harvest for anterior cervical fusion. Rates of complications and residual pain after ICBG harvest have reduced its utility as a source of autograft for spinal fusion. Less invasive options reduce morbidity, and trephine harvest is a simple technique, with low morbidity and perioperative and long-term pain. It has not been presented in the peer-reviewed spine literature, and the long-term experience has not been previously reported. Sixty-eight patients undergoing anterior cervical discectomy and fusion (ACDF), using allograft spacers, underwent ICBG harvest using a 3.5 mm trephine by a minimally invasive approach. Perioperative complications, pain incidence and Visual Analog Scale (VAS) were analyzed. Thirty consecutive patients were subsequently surveyed for long-term follow-up regarding pain, function, and satisfaction. No perioperative (0-6 weeks) graft-site complications (infection, hematoma, fracture, wound dehiscence) occurred. There were no reoperations, and no treatment required for graft-site complaints. No patient reported clinically significant perioperative pain (>3/10). At long-term follow-up (mean 45 months) no patients reported pain >3/10 at their graft-site, and the mean VAS was 0.20/10. No patient was impaired by graft-site symptoms, and all were satisfied with their graft harvest. Trephine graft harvest minimizes soft tissue dissection, periosteal elevation, and cortical disruption. Outcome using this technique eliminated perioperative complications in this study group, and reduced pain in both perioperative and long-term assessments. Compared to historical outcomes, trephine harvest provides sufficient graft with comparable or improved results relative to other minimally invasive approaches, and significantly improved relative to traditional open techniques. 4.


Point source characterization of per- and polyfluoroalkyl substances (PFASs) and extractable organofluorine (EOF) in freshwater and aquatic invertebrates.
Major point sources of per- and polyfluoroalkyl substances (PFASs) cause ubiquitous spread of PFASs in the environment. In this study, surface water and aquatic invertebrates at three Swedish sites impacted by PFAS point sources were characterized, using homologue, isomer and extractable organofluorine (EOF) profiling as well as estimation of bioaccumulation factors (BAFs) and mass discharge. Two sites were impacted by fire training (sites A and R) and the third by industrial runoff (site K). Mean Σ25PFASs concentration in water was 1920 ng L-1 at site R (n = 3), which was more than 20- and 10-fold higher than those from sites A and K, respectively. PFOS was the most predominant PFAS in all waters samples, constituting 29-79% of Σ25PFAS concentrations. Several branched isomers were detected and they substantially contributed to concentrations in surface water (e.g. 49-78% of ΣPFOS) and aquatic invertebrates (e.g. 15-28% of ΣPFOS). BAFs in the aquatic invertebrates indicated higher bioaccumulation for long chain PFASs and lower bioaccumulation for branched PFOS isomers compared to linear PFOS. EOF mass balance showed that Σ25target PFASs in water could explain up to 55% of EOF at site R. However, larger proportions of EOF (>92%) remained unknown in water from sites A and K. Mass discharges were for the first time estimated for EOF and revealed that high amounts of EOF (e.g. 8.2 g F day-1 at site A) could be transported by water to recipient water bodies relative to Σ25PFASs (e.g. 0.15 g day-1 at site A). Overall, we showed that composition profiling, BAFs and EOF mass balance can improve the characterization of PFASs around point sources.


Good-quality blastocysts derived from vacuolized morulas show reduced viability.
To study whether late spontaneous vacuolization on day 4 is an artefact or an alternate means of blastocele formation and to analyze its impact on pregnancy outcome and live birth. Prospective observational study. University teaching hospital. A total of 424 patients who fulfilled inclusion criteria were subgrouped according to the spontaneous vacuolization on day 4: Group 1 had all morulas affected, group 2 showed no signs of vacuoles, and group 3 was mixed (some day 4 embryos had vacuoles and others did not). Screening for the presence of vacuoles on day 4 and fresh single-blastocyst transfer. Morula and blastocyst scoring, utilization rate, pregnancy and live birth rates. Patients of group 1 had a reduced blastocyst formation rate on day 5 (P<.01) and significantly fewer good-quality blastocysts for usage (P<.05). In addition, pregnancy (P<.001) and live birth (P<.01) rate were significantly worse in group 1 compared with groups 2 and 3. Late onset of vacuolization around compaction stage is a negative predictor of blastocyst formation and outcome.


Secondary glaucoma after paediatric cataract surgery.
To determine the prevalence and risk factors associated with secondary glaucoma postcongenital cataract surgery. All children diagnosed as having congenital cataracts in a major children's hospital between 1985 and 2005 were included in a retrospective case series. Medical records of 423 eyes among 283 patients who underwent cataract surgery with or without intraocular lens implantation at age </=16 for congenital cataract were reviewed. The main outcome measure was presence or absence of secondary glaucoma and time to glaucoma postsurgery. The following risk factors were evaluated: age at cataract surgery, presence of systemic anomalies, microcornea, persistent hyperplastic primary vitreous (PHPV), primary capsulotomy/anterior vitrectomy, primary intraocular lens implantation, secondary membrane surgery and duration of postoperative observation. The statistical methods were the use of Kaplan-Meier survival analysis and Multivariate Cox hazards regression analysis. The mean follow-up was 6.3 (SD 5.0) years (median 4.6 years; range 0.5 to 20.3 years). Glaucoma developed in 36 of 234 patients (15.4%). Multivariate Cox proportional hazards regression analysis identified age less than 9 months at time of surgery (RR 2.9, 95% CI 1.3 to 7.7; p = 0.03), microcornea (RR 3.7, 95% CI 2.0 to 7.0; p<0.001), and follow-up time as important predictors of glaucoma. PHPV (RR 1.4, 95% CI 0.7 to 2.7; p = 0.41) and primary posterior capsulotomy/anterior vitrectomy (RR 2.2, 95% CI 0.9 to 5.5; p = 0.17) were not significantly associated with secondary glaucoma in the multivariate model. The mean time to glaucoma after congenital cataract surgery was 4.9 years (range 2 weeks to 16.8 years). Secondary glaucoma is an important sequela in patients who undergo surgery for congenital cataracts. It is imperative that these patients get lifelong surveillance, as glaucoma can occur years after the initial operation.


What do we know about rate-setting?
State regulation of hospital revenue is one approach to cost containment on which there is voluminous evaluative data from the "laboratory of the states"; yet these data are sufficiently complex and ambiguous that they are claimed by both proponents and opponents of rate-setting as support for their respective cases. This paper summarizes the evidence, concluding that mandatory rate-setting has generally constrained hospital costs where it has been implemented. However, were rate-setting established in additional states, it is not clear that comparable results would be realized. It is still less clear that rate-setting would constrain health care costs more than would increased competition and selective contracting.


Dynamic behaviors of vimentin induced by interaction with GlcNAc molecules.
The cytoskeleton protein vimentin is dramatically altered following pathological events such as fibrosis and tumorigenesis. Vimentin binds to multivalent N-acetylglucosamine (GlcNAc) molecules at the cell surface and interacts with O-linked β-GlcNAc proteins. Moreover, dying cells can be engulfed by neighboring cells through surface interactions between vimentin and many O-GlcNAc proteins in cell debris. Here, we show that vimentin was altered by its interaction with GlcNAc-bearing molecules such as GlcNAc-bearing polymers. The interaction with GlcNAc-bearing polymers promoted the cell surface recruitment of vimentin followed by the phosphorylation of vimentin serine 71 and the increase in tetrameric vimentin disassembled from vimentin filaments in HeLa cells. Moreover, it was found that GlcNAc-bearing polymers and O-GlcNAc proteins from dying cells promoted vimentin expression and cell migration in the Madin-Darby canine kidney and Michigan Cancer Foundation-7 cells. These results suggest that interactions between surface vimentin and GlcNAc molecules, including the O-GlcNAc proteins from dying cells, may play a pivotal role in vimentin expression and the migration of cancer cells. We propose new mechanisms of vimentin expression in cancer cells.


Visual-oculomotor interactions facilitate consolidation of perceptual learning.
Visual skill learning is commonly considered a manifestation of brain plasticity. Following encoding, consolidation of the skill may result in between-session performance gains. A great volume of studies have demonstrated that during the offline consolidation interval, the skill is susceptible to external inputs that modify the preformed representation of the memory, affecting future performance. However, while basic visual perceptual learning is thought to be mediated by sensory brain regions or their higher-order readout pathways, the possibility of visual-oculomotor interactions affecting the consolidation interval and reshaping visual learning remains uncharted. Motivated by findings mapping connections between oculomotor behavior and visual performance, we examined whether visual consolidation can be facilitated by visual-oculomotor interactions. To this aim, we paired reactivation of an oculomotor memory with consolidation of a typical visual texture discrimination task. Importantly, the oculomotor memory was encoded by learning of the pure motor component of the movement, removing visual cues. When brief reactivation of the oculomotor memory preceded the visual task, visual gains were substantially enhanced compared with those achieved by visual practice per se and were strongly related to the magnitude of oculomotor gains, suggesting that the brain utilizes oculomotor memory to enhance basic visual perception.


Stability of plant virus-based nanocarriers in gastrointestinal fluids.
Cowpea mosaic virus (CPMV) is a plant virus which is being extensively investigated as a drug delivery and vaccine nanocarrier for parenteral administration. However, to date little is known about the suitability of plant-based nanocarriers for oral delivery. In this study, the colloidal (i.e. aggregation), physical (i.e. denaturation) and chemical (i.e. digestion of the polypeptides) stability of CPMV and its empty virus-like particles (eVLPs) in conditions resembling the gastrointestinal fluids were evaluated. The nanoparticles were incubated in various simulated gastric and intestinal fluids and in pig gastric and intestinal fluids. CPMV and eVLPs had similar stabilities. In simulated gastric media, they were stable at pH ≥ 2.5. At lower pH destabilisation of the particle structure occurred, which, in turn, rendered the polypeptides extremely sensitive to pepsin digestion. However, both CPMV and eVLPs were stable in simulated intestinal fluids, in pig gastric fluids and in pig intestinal fluids. Thus CPMV, despite being a protein-based nanoparticle, was much more resistant to the harsh GI conditions than soluble proteins. Remarkably, both CPMV and eVLPs incubated in pig gastric and intestinal fluids were not subject to protein adsorption, with no formation of a detectable protein corona. The lack of a protein corona on CPMV and eVLP surfaces in GI fluids would imply that, if orally administered, these nanoparticles could maintain their native surface characteristics; thus, their biological interactions would remain predictable and unchanged. In summary, CPMV and eVLPs can be considered promising nanocarriers for applications requiring oral delivery, given their chemical, physical and colloidal stability and lack of protein adsorption from the environment in most of the tested conditions.


In vivo measurement of aromatase inhibition by letrozole (CGS 20267) in postmenopausal patients with breast cancer.
Thirteen postmenopausal women with advanced breast cancer were enrolled in an open randomized Phase I trial of a new p.o. active aromatase inhibitor, CGS 20267 (letrozole). The primary aim of the trial was to assess the impact of two doses of letrozole (0.5 and 2. 5 mg/day) on the peripheral aromatization of androstenedione to estrone. An in vivo isotopic technique was used to measure peripheral aromatization in each patient before treatment. The patients were then randomly assigned to one of the two doses, and measurements of aromatization were repeated after 6 weeks. At 0.5 mg and 2.5 mg/day, letrozole inhibited aromatization by 98.4% (97.3 to >99.1) and >98.9% (98.5 to >99.1; geometric means and ranges), respectively. Plasma estrogen levels were also measured before and during treatment. At the dose of 0.5 mg/day estrone and estradiol levels fell by 82.0% and 84.1% (geometric means), respectively. At the dose of 2.5 mg/ day, the estrogens fell by 80.8% and 68.1%, respectively. There were no significant differences between the doses in aromatase inhibition. No formal statistical analysis was performed on the estrogen data. Letrozole is therefore a highly effective inhibitor of aromatase, causing near complete inhibition of the enzyme in peripheral tissues at the doses investigated. The falls in estrogen levels were greater than those seen with earlier generation aromatase inhibitors.


High-speed electrooptic spectral scanning.
A spectral scanning technique is described which is capable of spectral scanning in the ultraviolet, visible, and infrared regions in times as short as fractions of a microsecond, with scan repetition rates of the order of a reciprocal scan time. The technique utilizes an electrooptic material to deflect the dispersed light in a monochromator past a fixed exit slit. Scan ranges up to two hundred times the theoretical resolution of the monochromator employed can be obtained in the visible spectral region. Results are presented for scans in the visible and infrared (2.7 microm) spectral regions using LiNbOa(3) as the electrooptic element.


Spectral characterization of laser-driven solid-based high harmonics in the coherent wake emission regime.
Laser-produced surface high-harmonic generation is an attractive source of extreme ultraviolet radiation due to its coherent properties and high peak power. By operating at subrelativistic laser intensities in the coherent wake emission regime, the harmonic spectrum was experimentally studied versus laser properties. At higher laser intensities (>10(17) W/cm(2)) a higher spectral cutoff was observed, with accompanying blueshifting and spectral broadening of the individual orders. A model based on an expanding critical surface provides qualitative agreement with the observations.


Cancer Patients' and Survivors' Perceptions of the Calm App: Cross-Sectional Descriptive Study.
There is a need for tools to decrease cancer patients' and survivors' long-term symptom burden. Complementary strategies, such as meditation, can accompany pharmacologic therapy to improve symptoms. Although support programs with targeted content have wider reach, higher adherence, and greater impact, there are no consumer-based meditation apps designed specifically for cancer. This study aimed to gather information to advise the development of a cancer-specific meditation app in a small convenience sample of cancer patients and survivors who currently use the Calm app. Adult cancer patients and survivors who are Calm users (N=82) were recruited through the Daily Calm Facebook page. Participants completed a Web-based survey related to Calm app use and satisfaction, interest in and ideas for a cancer-specific Calm app, and demographic characteristics. Open-ended responses were inductively coded. Participants were aged between 18 and 72 years (mean 48.60 years, SD 15.20), mostly female (77/82, 94%), white (65/79, 82%), and non-Hispanic (70/75, 93%), and reported using Calm at least 5 times per week (49/82, 60%). Although rates of satisfaction with current Calm components were high (65/82, 79%; and 51/81, 63%), only 49% (40/82) of participants used guided meditations that they felt specifically helped with their cancer-related symptoms and survivorship, and 40% (33/82) would prefer more cancer-related content, with guided meditations for cancer-specific anxieties (eg, fear of recurrence; n=15) and coping with strong emotions (n=12) being the most common suggestions. A majority of participants (51/82, 62%) reported that they would be interested in becoming a member of a Calm cancer community (eg, in-app discussion boards: 41/46, 89%; and social media communities: 35/42, 83%). Almost half of the participants (37/82, 45%) reported that they would benefit from features that tracked symptoms in concurrence with app usage, but respondents were divided on whether this information should be shared with health care providers through the app (49/82, 60% would share). Responses suggest ways in which the current Calm app could be adapted to better fit cancer patients' and survivors' needs and preferences, including adding more cancer-specific content, increasing the amount of content focusing on coping with strong emotions, developing communities for Calm users who are cancer patients and survivors, and including features that track cancer-related symptoms. Given differences in opinions about which features were desirable or would be useful, there is a clear need for future cancer-specific apps to be customizable (eg, ability to turn different features on or off). Although future research should address these topics in larger, more diverse samples, these data will serve as a starting point for the development of cancer-specific meditation app and provide a framework for evaluating their effects.


GaN-based light-emitting diodes with photonic crystals structures fabricated by porous anodic alumina template.
In this paper, we propose and demonstrate a convenient and flexible approach for preparation large-area of photonic crystals (PhCs) structures on the GaN-based LED chip. The highly-ordered porous anodic alumina (AAO) with pitch of wavelength scale was adopted as a selective dry etching mask for PhCs-pattern transfer. The PhCs with different pore depths were simultaneously formed on the entire surfaces of GaN-based LED chip including ITO, GaN surrounding contacts and the sidewall of the mesa by one-step reactive ion etching (RIE). The light output power improvement of PhCs-based GaN LED was achieved as high as 94% compared to that of the conventional GaN-based LED.


On the benefits of using multivariate analysis in mass spectrometric studies of combustion-generated aerosols.
The intricate chemistry of the carbonaceous particle surface layer (which drives their reactivity, environmental and health impacts) results in complex mass spectra. In this respect, detailed molecular-level analysis of combustion emissions may be challenging even with high-resolution mass spectrometry. Building on a recently proposed comprehensive methodology (encompassing all stages from sampling to data reduction), we propose herein a comparative analysis of soot particles produced by three different sources: a miniCAST standard generator, a laboratory diffusion flame and a single cylinder internal combustion engine. The surface composition is probed by either laser or secondary ion mass spectrometry. Two examples of multivariate analysis, Principal component analysis and hierarchical clustering analysis proved their efficiency in both identifying general trends and evidencing subtle differences that otherwise would remain unnoticed in the plethora of data generated during mass spectrometric analyses. Chemical information extracted from these multivariate statistical procedures contributes to a better understanding of fundamental combustion processes and also opens to practical applications such as the tracing of engine emissions.


Neural networks with local receptive fields and superlinear VC dimension.
Local receptive field neurons comprise such well-known and widely used unit types as radial basis function (RBF) neurons and neurons with center-surround receptive field. We study the Vapnik-Chervonenkis (VC) dimension of feedforward neural networks with one hidden layer of these units. For several variants of local receptive field neurons, we show that the VC dimension of these networks is superlinear. In particular, we establish the bound Omega(W log k) for any reasonably sized network with W parameters and k hidden nodes. This bound is shown to hold for discrete center-surround receptive field neurons, which are physiologically relevant models of cells in the mammalian visual system, for neurons computing a difference of gaussians, which are popular in computational vision, and for standard RBF neurons, a major alternative to sigmoidal neurons in artificial neural networks. The result for RBF neural networks is of particular interest since it answers a question that has been open for several years. The results also give rise to lower bounds for networks with fixed input dimension. Regarding constants, all bounds are larger than those known thus far for similar architectures with sigmoidal neurons. The superlinear lower bounds contrast with linear upper bounds for single local receptive field neurons also derived here.


Using mental mapping to unpack perceived cycling risk.
Cycling is the most energy-efficient mode of transport and can bring extensive environmental, social and economic benefits. Research has highlighted negative perceptions of safety as a major barrier to the growth of cycling. Understanding these perceptions through the application of novel place-sensitive methodological tools such as mental mapping could inform measures to increase cyclist numbers and consequently improve cyclist safety. Key steps to achieving this include: (a) the design of infrastructure to reduce actual risks and (b) targeted work on improving safety perceptions among current and future cyclists. This study combines mental mapping, a stated-preference survey and a transport infrastructure inventory to unpack perceptions of cycling risk and to reveal both overlaps and discrepancies between perceived and actual characteristics of the physical environment. Participants translate mentally mapped cycle routes onto hard-copy base-maps, colour-coding road sections according to risk, while a transport infrastructure inventory captures the objective cycling environment. These qualitative and quantitative data are matched using Geographic Information Systems and exported to statistical analysis software to model the individual and (infra)structural determinants of perceived cycling risk. This method was applied to cycling conditions in Galway City (Ireland). Participants' (n=104) mental maps delivered data-rich perceived safety observations (n=484) and initial comparison with locations of cycling collisions suggests some alignment between perception and reality, particularly relating to danger at roundabouts. Attributing individual and (infra)structural characteristics to each observation, a Generalised Linear Mixed Model statistical analysis identified segregated infrastructure, road width, the number of vehicles as well as gender and cycling experience as significant, and interactions were found between individual and infrastructural variables. The paper concludes that mental mapping is a highly useful tool for assessing perceptions of cycling risk with a strong visual aspect and significant potential for public participation. This distinguishes it from more traditional cycling safety assessment tools that focus solely on the technical assessment of cycling infrastructure. Further development of online mapping tools is recommended as part of bicycle suitability measures to engage cyclists and the general public and to inform 'soft' and 'hard' cycling policy responses.


ELECTROCHEMICAL COUPLING IN POTENTIATION OF MUSCULAR CONTRACTION.
Diverse potentiators of contraction have basically identical, activestate mechanical effects, but act by different membrane-mediated electromechanical coupling mechanisms. The falling phase of the action potential is greatly prolonged by Zn(2+) (0.1 mM) and UO(2)(2+) (0.5 to 1 microM), neither of which affects the mechanical threshold. Caffeine (1 mM), like the lyotropic anions, acts conversely. Thus changes in the duration and mechanical threshold of the action potential determine independent electromechanical coupling processes which can act individually, or conjointly in the action of other potentiators, in determining the duration of the active state and thus the potentiation of twitch tension.


Phase separation of binary mixtures induced by soft centrifugal fields.
We use the model system ethanol-dodecane to demonstrate that giant critical fluctuations induced by easily accessible weak centrifugal fields as low as 2000g can be observed above the miscibility gap of a binary liquid mixture. Moreover, several degrees above the phase transition, i.e. in the one-phase region, strong gradients of ethanol concentration occur upon centrifugation. In this case, the standard interpretation of sedimentation equilibrium in the analytical ultracentrifuge (AUC) yields an apparent molar mass of ethanol three orders of magnitude higher than the real value. Notably, these composition gradients have no influence on the distribution gradient of solutes such as dyes like Nile red. The thick opaque interphase formed upon centrifugation does not appear as the commonly observed sharp meniscus, but as a turbidity zone, similar to critical opalescence. This layer is a few millimeters thick and separates two fluids with low compositional gradients. All these effects can be qualitatively understood and explained using the Flory-Huggins solution model coupled to classical density functional theory (DFT). In this domain hetero-phase fluctuations can be triggered by gravity even far from the critical point. Taking into account Jean Perrin's approach to external fields in colloids, a self-consistent definition of the Flory effective volume and an explicit calculation of the total free energy per unit volume is possible.


Erythropoiesis-stimulating Agents in Critically Ill Trauma Patients: A Systematic Review and Meta-analysis.
To perform a meta-analysis of all relevant randomized controlled trials assessing the effect of erythropoiesis-stimulating agents (ESAs) in critically ill trauma patients. ESAs have effects beyond erythropoiesis. The administration of the ESA epoetin alfa to critically ill trauma patients has been associated with a reduction in mortality. We performed a systematic review and meta-analysis with trial sequential analysis. We searched Medline, Medline in Process, and other nonindexed citations, EMBASE, and the Cochrane Database from inception until September 9, 2015, for randomized controlled trials comparing ESAs to placebo (or no ESA). We identified 9 eligible studies that randomly assigned 2607 critically ill patients after trauma to an ESA or placebo (or no ESA). Compared with placebo (or no ESA), ESA therapy was associated with a substantial reduction in mortality [risk ratio (RR) 0.63, 95% confidence interval (CI) 0.49-0.79, P = 0.0001, I = 0%). In patients with traumatic brain injury, ESA therapy did not increase the number of patients surviving with moderate disability or good recovery (RR 1.00, 95% CI 0.88-1.15, P = 0.95, I = 0%). With the dosing regimens employed in the included studies, ESA therapy did not increase the risk of lower limb proximal deep venous thrombosis (RR 0.97, 95% CI 0.72-1.29, P = 0.78, I = 0%). The administration of ESAs to critically ill trauma patients is associated with a significant improvement in mortality without an increase in the rate of lower limb proximal deep venous thrombosis. Given the worldwide public health significance of these findings research to validate or refute them is required.


Ecotoxicological risk assessment of chemical pollution in four Iberian river basins and its relationship with the aquatic macroinvertebrate community status.
Ecotoxicological risk assessment of chemical pollution in four Iberian river basins (Llobregat, Ebro, Júcar and Guadalquivir) was performed. The data set included more than 200 emerging and priority compounds measured at 77 sampling sites along four river basins studied. The toxic units (TU) approach was used to assess the risk of individual compounds and the concentration addition model (CA) to assess the site specific risk. Link between chemical pollution and aquatic macroinvertebrate communities in situ was examined by using four biological indexes; SPEAR ("Species at Risk Index") as the indicator of decline of sensitive species in relation to general organic (SPEARorganic) and pesticides (SPEARpesticides) pollution; and Shannon and Margalef biodiversity indexes. The results of the study suggested that organic chemicals posed the risk of acute effects at 42% of the sampling sites and the risk of chronic effects at all the sites. Metals posed the acute risk at 44% of the sites. The main drivers of the risk were mainly pesticides and metals. However, several emerging contaminants (e.g. the antidepressant drug sertraline and the disinfectant triclosan) were contributing to the chronic effects risk. When risk associated with metals and organic chemicals was compared, the latter dominated in 2010, mainly due to the presence of highly toxic pesticides, while metals did in 2011. Compounds that are not regulated on the European level were posing the risk of chronic effects at 23% of the sites. The decline of sensitive macroinvertebrate taxa expressed in terms of SPEAR index was correlated with the increase of toxic stress related to organic compounds Biodiversity indexes were negatively correlated with the metals and the urban land use type in the catchment.


Molecular phenotyping of maternally mediated parallel adaptive divergence within Rana arvalis and Rana temporaria.
When similar selection acts on the same traits in multiple species or populations, parallel evolution can result in similar phenotypic changes, yet the underlying molecular architecture of parallel phenotypic divergence can be variable. Maternal effects can influence evolution at ecological timescales and facilitate local adaptation, but their contribution to parallel adaptive divergence is unclear. In this study, we (i) tested for variation in embryonic acid tolerance in a common garden experiment and (ii) used molecular phenotyping of egg coats to investigate the molecular basis of maternally mediated parallel adaptive divergence in two amphibian species (Rana arvalis and Rana temporaria). Our results on three R. arvalis and two R. temporaria populations show that adaptive divergence in embryonic acid tolerance is mediated via maternally derived egg coats in both species. We find extensive polymorphism in egg jelly coat glycoproteins within both species and that acid-tolerant clutches have more negatively charged egg jelly - indicating that the glycosylation status of the jelly coat proteins is under divergent selection in acidified environments, likely due to its impact on jelly water balance. Overall, these data provide evidence for parallel mechanisms of adaptive divergence in two species. Our study highlights the importance of studying intraspecific molecular variation in egg coats and, specifically, their glycoproteins, to increase understanding of underlying forces maintaining variation in jelly coats.


DNA restriction-fragment variation in the gene family encoding high molecular weight (HMW) glutenin subunits of wheat.
Restriction enzyme digests of DNA from nullisomic-tetrasomic and intervarietal chromosome substitution lines of wheat were probed with a high molecular weight (HMW) glutenin cDNA. Three restriction endonucleases were used to investigate restriction-fragment differences among five wheat varieties. The results suggest that the hybridizing fragments contain single gene copies and permit the identification of the subunit encoded by each gene. Restriction-fragment variation associated with previously established allelic differences between varieties was observed. Also, there is a clear relationship between the electrophoretic mobility of a HMW subunit and the length of the central repetitive section of the gene encoding it. These results are discussed with reference to the evolution of the HMW glutenin gene family and the uses of restriction-fragment variation in plant breeding and genetics.


Countertransference in Swedish psychotherapists: testing the factor structure of the Therapist Response Questionnaire.
Questionnaires need testing of reliability and factor structure before clinical use or research in new languages or cultures. The aim of this study was to evaluate the Therapist Response Questionnaire (TRQ) in Sweden compared to corresponding factor analyses in USA and Italy. A national sample of psychotherapists (N=242) registered their countertransference with a single client using TRQ. The data were analyzed with confirmatory factor analysis (CFA) to test factor structures from previous studies, and exploratory factor analysis (EFA). The CFA did not verify the factor structure from the previous studies. The EFA extracted seven factors as the best solution: Helpless/Inadequate, Overwhelmed/Disorganized, Hostile/Angry, Parental/Protective, Disengaged, Special/Overinvolved, Sexualized. Analysis of convergent validity indicated that five of these could be considered equivalent to factors in the previous studies, and the remaining two were conceptually related to corresponding factors. Even though the factor structure was not confirmed by the CFA, the concordance was large, indicating a reliable self-report instrument with promising validity for measurement of complex aspects of countertransference. Common countertransference themes can inform psychotherapy supervision and education, give feedback to the therapist, and lay ground for a taxonomy for therapist reactions and feelings.


Reaction-diffusion in a growing 3D domain of skin scales generates a discrete cellular automaton.
We previously showed that the adult ocellated lizard skin colour pattern is effectively generated by a stochastic cellular automaton (CA) of skin scales. We additionally suggested that the canonical continuous 2D reaction-diffusion (RD) process of colour pattern development is transformed into this discrete CA by reduced diffusion coefficients at the borders of scales (justified by the corresponding thinning of the skin). Here, we use RD numerical simulations in 3D on realistic lizard skin geometries and demonstrate that skin thickness variation on its own is sufficient to cause scale-by-scale coloration and CA dynamics during RD patterning. In addition, we show that this phenomenon is robust to RD model variation. Finally, using dimensionality-reduction approaches on large networks of skin scales, we show that animal growth affects the scale-colour flipping dynamics by causing a substantial decrease of the relative length scale of the labyrinthine colour pattern of the lizard skin.


Hormonal regulation of hepatic gluconeogenesis: influence of age and training.
The contributions of three major gluconeogenic regulators, glucagon (10(-7) M), alpha-adrenergic agonist phenylephrine (10(-5) M), and beta-agonist isoproterenol (10(-5) M) to hepatic glucose synthesis in liver slices from Fischer 344 rats were examined in relation to age and endurance training. Young (4 mo), middle-aged (12 mo), and old (22 mo) male Fischer 344 rats (n = 66) were divided into trained or sedentary groups. Trained animals were run 10 wk on a treadmill at 75% of maximal capacity, 1 h/day, 5 days/wk. Animals were killed at rest, and sections of liver were removed and sliced in a tissue microtome. Slices were incubated in L-[U-14C]lactic acid, Ringer solution, and one of the aforementioned gluconeogenic regulators. Rates of lactate incorporation into glucose and glycogen were significantly greater in young compared with old animals for all three regulators in both trained and untrained animals. Training elicited a 35, 52, and 63% improvement in lactate incorporation into glucose compared with untrained when the livers of young (16.9 +/- 1.2 vs. 10.9 +/- 1.1 mumol.g protein-1.min-1), middle-aged (12.8 +/- 1.3 vs. 6.1 +/- 1.2 mumol.g protein-1.min-1), and old (11.2 +/- 1.1 vs. 4.1 +/- 0.6 mumol.g protein-1.min-1) animals, respectively, were incubated in glucagon. Rates with phenylephrine followed a similar pattern to that with glucagon across age and training, but absolute rates were significantly lower. No training effect in gluconeogenic rate was found when liver was incubated in the presence of isoproterenol. It is concluded that the gluconeogenic capacity of liver declines with age regardless of the gluconeogenic regulator and that training was able to partially offset age-related declines in glucagon-stimulated and alpha-receptor-mediated gluconeogenesis.


Differential Effect of SdhB Gene Mutations on the Sensitivity to SDHI Fungicides in Botrytis cinerea.
Succinate dehydrogenase inhibiting (SDHI) fungicides constitute a relatively novel fungicide group used for gray mold control caused mainly by Botrytis cinerea. Shortly after registration, resistance was observed in fungal populations that correlated with several mutations in the succinate dehydrogenase complex (complex II). In the current study, 30 B. cinerea isolates possessing five different mutations at three different codons of SdhB (P225F, N230I, and H272L/R/Y) were characterized for their sensitivities to eight SDHI fungicides. The results show different sensitivities and cross-resistance patterns between structurally different SDHIs. P225F mutants were resistant in vitro to all SDHIs tested. Similarly, isolates possessing the H272L mutation were highly resistant to boscalid but showed low to moderate levels of resistance to other SDHIs. The N230I mutants were moderately resistant to boscalid, fluopyram, and fluxapyroxad and showed low resistance levels to isopyrazam, bixafen, fenfuram, benodanil, and carboxin. The H272R mutants showed moderate levels of resistance to boscalid and low resistance levels to isopyrazam, fenfuram, and carboxin but remained sensitive to fluopyram, bixafen, fluxapyroxad, and benodanil. Similarly, the H272Y showed moderate levels of resistance to boscalid and very low resistance levels to isopyrazam, bixafen, fenfuram, and carboxin but showed increased sensitivity to benodanil and fluopyram. Boscalid provided moderate to high control of H272R/Y and N230I mutants in detached fruit assays but provided little control against the H272L and P225F mutants. In contrast, fluopyram controlled H272R/Y mutants and provided moderate levels of control toward H272L, N230I, and P225F mutants. Our findings suggest that sensitivity to SDHIs may vary greatly, dependent on the point mutation in the sdhb subunit.


Effect of local TGF-beta1 and IGF-1 release on implant fixation: comparison with hydroxyapatite coating: a paired study in dogs.
Hydroxyapatite (HA) coating stimulates the osseointegration of cementless orthopedic implants. Recently, locally released osteogenic growth factors have also been shown experimentally to stimulate osseointegration so that bone fills gaps around orthopedic implants. Here, we have compared the effect of local release of TGF-beta1 and IGF-1 with that of hydroxyapatite coating on implant fixation. Weight-bearing implants with a 0.75-mm surrounding gap were inserted bilaterally in the knees of 10 dogs. Growth factors were incorporated in a biodegradable poly(D,L-lactide) coating on porous coated titanium implants. Plasma-sprayed HA implants served as controls. The dogs were killed at 4 weeks and the implants were evaluated by mechanical push-out test and by histomorphometry. There was no difference in any of the mechanical parameters. Bone ongrowth was 3-fold higher for HA-coated implants (p < 0.001). For growth factor-coated implants, bone volume was 26% higher in the inner half of the gap and 28% higher in the outer half compared to HA (p < 0.03). The mechanical fixation of porous-coated titanium implants with local growth factor release is comparable to that of HA coating. While HA mainly stimulated bone ongrowth, local release of TGF-beta1 and IGF-1 stimulated gap healing.


Drug metabolism in human brain: high levels of cytochrome P4503A43 in brain and metabolism of anti-anxiety drug alprazolam to its active metabolite.
Cytochrome P450 (P450) is a super-family of drug metabolizing enzymes. P450 enzymes have dual function; they can metabolize drugs to pharmacologically inactive metabolites facilitating their excretion or biotransform them to pharmacologically active metabolites which may have longer half-life than the parent drug. The variable pharmacological response to psychoactive drugs typically seen in population groups is often not accountable by considering dissimilarities in hepatic metabolism. Metabolism in brain specific nuclei may play a role in pharmacological modulation of drugs acting on the CNS and help explain some of the diverse response to these drugs seen in patient population. P450 enzymes are also present in brain where drug metabolism can take place and modify therapeutic action of drugs at the site of action. We have earlier demonstrated an intrinsic difference in the biotransformation of alprazolam (ALP) in brain and liver, relatively more alpha-hydroxy alprazolam (alpha-OHALP) is formed in brain as compared to liver. In the present study we show that recombinant CYP3A43 metabolizes ALP to both alpha-OHALP and 4-hydroxy alprazolam (4-OHALP) while CYP3A4 metabolizes ALP predominantly to its inactive metabolite, 4-OHALP. The expression of CYP3A43 mRNA in human brain samples correlates with formation of relatively higher levels of alpha-OH ALP indicating that individuals who express higher levels of CYP3A43 in the brain would generate larger amounts of alpha-OHALP. Further, the expression of CYP3A43 was relatively higher in brain as compared to liver across different ethnic populations. Since CYP3A enzymes play a prominent role in the metabolism of drugs, the higher expression of CYP3A43 would generate metabolite profile of drugs differentially in human brain and thus impact the pharmacodynamics of psychoactive drugs at the site of action.