Identification and characterization of a Ste20-like kinase in Artemia and its role in the developmental regulation and resistance to environmental stress.
To adapt to extreme environments, the crustacean Artemia has evolved two alternative reproductive pathways. During ovoviviparous (direct) development, nauplius larvae are produced. In contrast, Artemia females release encysted diapause embryos (cysts) via the oviparous pathway. To date, the cellular mechanisms that regulate stress resistance of Artemia remain largely unknown. Ste20-like kinase (SLK) participates in multiple biological processes, including stress responses, apoptosis, and cell cycle progression. We isolated and characterized a member of the SLK superfamily termed ArSLK from Artemia parthenogenetica. The ArSLK gene is transcribed throughout both ovoviviparous and oviparous development; however, the protein is located mainly in the nuclei of stress-resistant diapause cysts, unlike the nauplii and nauplius-destined embryos where it is cytoplasmic. Interestingly, exposure of nauplii to heat shock, acidic pH, and UV irradiation induced the translocation of ArSLK from cytoplasm to nucleus. This translocation was reversed following stress removal. Moreover, under physiologically-stressful conditions, the nauplius larvae produced by adults after gene knockdown of endogenous ArSLK by RNAi, lost the ability of free-swimming much earlier than those of control larvae from females injected with GFP dsRNA. Taken together, this study demonstrated that trafficking of ArSLK between the cytoplasm and the nucleus participates in regulating the stress resistance of Artemia. Our findings may provide significant insight into the functions of members of the SLK superfamily.


Remarkably low affinity of CD4/peptide-major histocompatibility complex class II protein interactions.
The αβ T-cell coreceptor CD4 enhances immune responses more than 1 million-fold in some assays, and yet the affinity of CD4 for its ligand, peptide-major histocompatibility class II (pMHC II) on antigen-presenting cells, is so weak that it was previously unquantifiable. Here, we report that a soluble form of CD4 failed to bind detectably to pMHC II in surface plasmon resonance-based assays, establishing a new upper limit for the solution affinity at 2.5 mM. However, when presented multivalently on magnetic beads, soluble CD4 bound pMHC II-expressing B cells, confirming that it is active and allowing mapping of the native coreceptor binding site on pMHC II. Whereas binding was undetectable in solution, the affinity of the CD4/pMHC II interaction could be measured in 2D using CD4- and adhesion molecule-functionalized, supported lipid bilayers, yielding a 2D Kd of ∼5,000 molecules/μm(2) This value is two to three orders of magnitude higher than previously measured 2D Kd values for interacting leukocyte surface proteins. Calculations indicated, however, that CD4/pMHC II binding would increase rates of T-cell receptor (TCR) complex phosphorylation by threefold via the recruitment of Lck, with only a small, 2-20% increase in the effective affinity of the TCR for pMHC II. The affinity of CD4/pMHC II therefore seems to be set at a value that increases T-cell sensitivity by enhancing phosphorylation, without compromising ligand discrimination.


Virology of hepatitis C virus.
Hepatitis C virus (HCV) has been identified as the main causative agent of posttransfusion non-A, non-B hepatitis. Through recently developed diagnostic assays, routine serologic screening of blood donors has prevented most cases of posttransfusion hepatitis. The purpose of this paper is to comprehensively review current information regarding the virology of HCV. Recent findings on the genome organization, its relationship to other viruses, the replication of HCV ribonucleic acid, HCV translation, and HCV polyprotein expression and processing are discussed. Also reviewed are virus assembly and release, the variability of HCV and its classification into genotypes, the geographic distribution of HCV genotypes, and the biologic differences between HCV genotypes. The assays used in HCV genotyping are discussed in terms of reliability and consistency of results, and the molecular epidemiology of HCV infection is reviewed. These approaches to HCV epidemiology will prove valuable in documenting the spread of HCV in different risk groups, evaluating alternative (nonparenteral) routes of transmission, and in understanding more about the origins and evolution of HCV.


Evaluation of right and left ventricular function using speckle tracking echocardiography in patients with arrhythmogenic right ventricular cardiomyopathy and their first degree relatives.
The identification of right ventricular abnormalities in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC) in early stages is still difficult. The aim of this study was to investigate if longitudinal strain based on speckle tracking can detect subtle right (RV) or left ventricular (LV) dysfunction as an early sign of ARVC. Seventeen male patients, fulfilling Task force criteria for ARVC, 49 (32-70) years old, nineteen male first degree relatives 29 (19-73) y.o. and twenty-two healthy male volunteers 36 (24-66) y.o participated in the study. Twelve-lead and signal-averaged electrocardiograms were recorded. All subjects underwent echocardiography. LV and RV diameters, peak systolic velocity from tissue Doppler and longitudinal strain based on speckle tracking were measured from the basal and mid segments in both ventricles. RV longitudinal strain measurement was successful in first degree relatives and controls (95 resp. 86%) but less feasible in patients (59%). Results were not systematically different between first degree relatives and controls. Using discriminant analysis, we then developed an index based on echocardiographic parameters. All normal controls had an index<l while patients with abnormal ventricles had an index between 1-4. Some of the first degree relatives deviated from the normal pattern. Longitudinal strain of LV and RV segments was significantly lower in patients than in relatives and controls. An index was developed incorporating dimensional and functional echocardiographic parameters. In combination with genetic testing this index might help to detect early phenotype expression in mutation carriers.


In vitro culture of epithelial cells from the caput, corpus, and cauda epididymis of Sus domesticus.
This work describes a protocol to culture epididymal epithelial cells from the caput, corpus, and cauda regions of Sus domesticus. Epididymal epithelial fragments were obtained by dissection and enzymatic digestion with collagenase. About 30 epididymal fragments from each epididymal region were cultured in 24-well culture plates with supplemented RPMI-1640 medium at 37 degrees C, 5% CO2 in air, and 100% humidity. A confluent monolayer of polygonal and tightly packed epithelioid cells from the three epididymal regions was obtained after 12-16 days in culture and maintained in vitro for more than 60 days. The proportion of epididymal epithelial cells in these cultures was assessed by immunofluorescent staining for cytokeratins. Throughout the 2 months of culture, about 80% of the cells were cytokeratin-positive. Electron microscopy observations indicated that cultured cells from caput, corpus, and cauda epididymal regions were tightly adhered to each other by junctional complexes and that stereocilia were present in their apical membranes. Moreover, the presence of an extensive rough endoplasmic reticulum, Golgi apparatus and numerous vesicles in the cytoplasm suggested that cultured cells maintained secretory and absorptive activities. These results show that the epididymal epithelial cells in culture from S. domesticus retain some fundamental features that characterize the epididymal epithelium in the intact organ. This system might be a valuable tool for studying the mechanism of sperm maturation in vitro, including epididymal cell secretions and the analysis of regional differences.


Andean cutaneous leishmaniasis (Andean-CL, uta) in Peru and Ecuador: the causative Leishmania parasites and clinico-epidemiological features.
This study provides comprehensive information on the past and current status of the Andean cutaneous leishmaniasis (Andean-CL, uta) in Peru and Ecuador, mainly focusing on the causative Leishmania parasites and clinico-epidemiological features. Available information and data including our unpublished works were analyzed thoroughly. Endemic regions of the Andean-CL (uta) in Peru run from the north Piura/Cajamarca to the south Ayacucho at a wide range of the Pacific watersheds of the Andes through several departments, while in Ecuador those exist at limited and spotted areas in the country's mid-southwestern two provinces, Azuay and Chimborazo. The principal species of the genus Leishmania are completely different at subgenus level, L. (Viannia) peruviana in Peru, and L. (Leishmania) mexicana and L. (L.) major-like (infrequent occurrence) in Ecuador. The Peruvian uta is now prevalent in different age and sex groups, being not clearly defined as found in the past. The precise reasons are not known and should be elucidated further, though probable factors, such as emergence of other Leishmania parasites, non-immune peoples' migration into the areas, etc., were discussed briefly in the text. The Andean-CL cases in Ecuador are more rural than before, probably because of a rapid development of the Leishmania-positive communities and towns, and the change of life-styles of the inhabitants, including newly constructed houses and roads in the endemic areas. Such information is helpful for future management of the disease, not only for Leishmania-endemic areas in the Andes but also for other endemic areas.


Interdigitated aluminium and titanium sensors for assessing epithelial barrier functionality by electric cell-substrate impedance spectroscopy (ECIS).
Electric cell-substrate impedance spectroscopy (ECIS) enables non-invasive and continuous read-out of electrical parameters of living tissue. The aim of the current study was to investigate the performance of interdigitated sensors with 50 μm electrode width and 50 μm inter-electrode distance made of gold, aluminium, and titanium for monitoring the barrier properties of epithelial cells in tissue culture. At first, the measurement performance of the photolithographic fabricated sensors was characterized by defined reference electrolytes. The sensors were used to monitor the electrical properties of two adherent epithelial barrier tissue models: renal proximal tubular LLC-PK1 cells, representing a normal functional transporting epithelium, and human cervical cancer-derived HeLa cells, forming non-transporting cancerous epithelial tissue. Then, the impedance spectra obtained were analysed by numerically fitting the parameters of the two different models to the measured impedance spectrum. Aluminium sensors proved to be as sensitive and consistent in repeated online-recordings for continuous cell growth and differentiation monitoring as sensors made of gold, the standard electrode material. Titanium electrodes exhibited an elevated intrinsic ohmic resistance in comparison to gold reflecting its lower electric conductivity. Analysis of impedance spectra through applying models and numerical data fitting enabled the detailed investigation of the development and properties of a functional transporting epithelial tissue using either gold or aluminium sensors. The result of the data obtained, supports the consideration of aluminium and titanium sensor materials as potential alternatives to gold sensors for advanced application of ECIS spectroscopy.


Intensity-based 2-D-3-D registration of cerebral angiograms.
We propose a new method for aligning three-dimensional (3-D) magnetic resonance angiography (MRA) with 2-D X-ray digital subtraction angiograms (DSA). Our method is developed from our algorithm to register computed tomography volumes to X-ray images based on intensity matching of digitally reconstructed radiographs (DRRs). To make the DSA and DRR more similar, we transform the MRA images to images of the vasculature and set to zero the contralateral side of the MRA to that imaged with DSA. We initialize the search for a match on a user defined circular region of interest. We have tested six similarity measures using both unsegmented MRA and three segmentation variants of the MRA. Registrations were carried out on images of a physical neuro-vascular phantom and images obtained during four neuro-vascular interventions. The most accurate and robust registrations were obtained using the pattern intensity, gradient difference, and gradient correlation similarity measures, when used in conjunction with the most sophisticated MRA segmentations. Using these measures, 95% of the phantom start positions and 82% of the clinical start positions were successfully registered. The lowest root mean square reprojection errors were 1.3 mm (standard deviation 0.6) for the phantom and 1.5 mm (standard deviation 0.9) for the clinical data sets. Finally, we present a novel method for the comparison of similarity measure performance using a technique borrowed from receiver operator characteristic analysis.


Executive function in phenylketonuria (PKU): Insights from the Behavior Rating Inventory of Executive Function (BRIEF) and a large sample of individuals with PKU.
Previous research has documented executive function (EF) impairments in individuals with early treated phenylketonuria (ETPKU). It remains unclear, however, whether some aspects of EF may be more affected than others. A number of factors, including small sample sizes and variability in EF tasks, have likely contributed to past mixed findings. The present objective was to elucidate further the EF profile associated with ETPKU, particularly as it relates to report-based assessment of EF. Data from 286 individuals (5-48 years of age) with ETPKU on the child and adult versions of the Behavior Rating Inventory of Executive Function (BRIEF), a well-established report-based assessment tool, were analyzed. The Working Memory scale showed the largest effect size in both young and older ETPKU samples, with 19% of children and 29% of adults scoring in the "abnormally elevated" range. In addition, EF impairment appeared more general (i.e., affecting more domains) in the adult sample as compared to the child sample. Exploratory analyses also suggested that the presence/absence of overall impairment on the BRIEF among our ETPKU participants could be predicted based on a small subset of items. A 10-item subset showed total classification accuracy values of 90% and above for both groups. Working memory represents an aspect of EF that appears to be particularly affected in individuals with ETPKU. Findings also provide preliminary support of the viability for the development and/or adoption of an abbreviated screening measure for EF difficulties in children and adults with ETPKU. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


The use of mystery shopping for quality assurance evaluations of HIV/STI testing sites offering services to young gay and bisexual men.
Young men who have sex with men (YMSM) are at increased risk for HIV and STI infection. While encouraging HIV and STI testing among YMSM remains a public health priority, we know little about the cultural competency of providers offering HIV/STI tests to YMSM in public clinics. As part of a larger intervention study, we employed a mystery shopper methodology to evaluate the LGBT cultural competency and quality of services offered in HIV and STI testing sites in Southeast Michigan (n = 43).We trained and deployed mystery shoppers (n = 5) to evaluate the HIV and STI testing sites by undergoing routine HIV/STI testing. Two shoppers visited each site, recording their experiences using a checklist that assessed 13 domains, including the clinic's structural characteristics and interactions with testing providers. We used the site scores to examine the checklist's psychometric properties and tested whether site evaluations differed between sites only offering HIV testing (n = 14) versus those offering comprehensive HIV/STI testing (n = 29). On average, site scores were positive across domains. In bivariate comparisons by type of testing site, HIV testing sites were more likely than comprehensive HIV/STI testing clinics to ascertain experiences of intimate partner violence, offer action steps to achieve safer sex goals, and provide safer sex education. The developed checklist may be used as a quality assurance indicator to measure HIV/STI testing sites' performance when working with YMSM. Our findings also underscore the need to bolster providers' provision of safer sex education and behavioral counseling within comprehensive HIV/STI testing sites.


Eccentric contraction: unraveling mechanisms of force enhancement and energy conservation.
During the past century, physiologists have made steady progress in elucidating the molecular mechanisms of muscle contraction. However, this progress has so far failed to definitively explain the high force and low energy cost of eccentric muscle contraction. Hypotheses that have been proposed to explain increased muscle force during active stretch include cross-bridge mechanisms, sarcomere and half-sarcomere length non-uniformity, and engagement of a structural element upon muscle activation. The available evidence suggests that force enhancement results from an interaction between an elastic element in muscle sarcomeres, which is engaged upon activation, and the cross-bridges, which interact with the elastic elements to regulate their length and stiffness. Similarities between titin-based residual force enhancement in vertebrate muscle and twitchin-based 'catch' in invertebrate muscle suggest evolutionary homology. The winding filament hypothesis suggests plausible molecular mechanisms for effects of both Ca(2+) influx and cross-bridge cycling on titin in active muscle. This hypothesis proposes that the N2A region of titin binds to actin upon Ca(2+) influx, and that the PEVK region of titin winds on the thin filaments during force development because the cross-bridges not only translate but also rotate the thin filaments. Simulations demonstrate that a muscle model based on the winding filament hypothesis can predict residual force enhancement on the descending limb of the length-tension curve in muscles during eccentric contraction. A kinematic model of titin winding based on sarcomere geometry makes testable predictions about titin isoforms in different muscles. Ongoing research is aimed at testing these predictions and elucidating the biochemistry of the underlying protein interactions.


Ethanol is self-administered into the nucleus accumbens shell, but not the core: evidence of genetic sensitivity.
A previous study indicated that selectively bred alcohol-preferring (P) rats self-administered ethanol (EtOH) directly into the posterior ventral tegmental area at lower concentrations than Wistar rats. The present study was undertaken to determine involvement of the nucleus accumbens (Acb) with EtOH reinforcement, and a relationship between genetic selection for high alcohol preference and sensitivity of the Acb to the reinforcing effects of EtOH. Adult P and Wistar rats were assigned to groups that self-infused 0 to 300 mg% EtOH into the Acb shell (AcbSh) or Acb Core (AcbC). Rats were placed into 2-lever (active and inactive) operant chambers and given EtOH for the first 4 sessions (acquisition), artificial cerebrospinal fluid (aCSF) for sessions 5 and 6 (extinction), and EtOH again in session 7 (reinstatement). Responding on the active lever produced a 100-nl injection of the infusate. Alcohol-preferring rats self-infused 75 to 300 mg% EtOH, whereas Wistar rats reliably self-infused 100 and 300 mg% EtOH into the AcbSh. Both P and Wistar rats reduced responding on the active lever when aCSF was substituted for EtOH, and reinstated responding in session 7 when EtOH was restored. EtOH was not self-infused into the AcbC by P or Wistar rats. The present results indicate that the AcbSh, but not AcbC, is a neuroanatomical structure that mediates the reinforcing actions of EtOH. The data also suggest that, compared to Wistar rats, the AcbSh of P rats is more sensitive to the reinforcing effects of EtOH.


Ability of cytosolic malate dehydrogenase and lactate dehydrogenase to increase the ratio of NADPH to NADH oxidation by cytosolic glycerol-3-phosphate dehydrogenase.
At the normal pH of the cytosol (7.0 to 7.1) and in the presence of physiological (1.0 mM) levels of free Mg2+, the Vmax of the NADPH oxidation is only slightly lower than the Vmax of NADH oxidation in the cytosolic glycerol-3-phosphate dehydrogenase (E.C. 220.127.116.11) reaction. Under these conditions physiological (30 microM) levels of cytosolic malate dehydrogenase (E.C. 126.96.36.199) inhibited oxidation of 20 microM NADH but had no effect on oxidation of 20 microM NADPH by glycerol-3-phosphate dehydrogenase. Consequently malate dehydrogenase increased the ratio of NADPH to NADH oxidation of glycerol-3-phosphate dehydrogenase. On the basis of the measured KD of complexes between malate dehydrogenase and these reduced pyridine nucleotides, and their Km in the glycerol-3-phosphate dehydrogenase reactions, it could be concluded that malate dehydrogenase would have markedly inhibited NADPH oxidation and inhibited NADH oxidation considerably more than observed if its only effect were to decrease the level of free NADH or NADPH. This indicates that due to the opposite chiral specificity of the two enzymes with respect to reduced pyridine nucleotides, complexes between malate dehydrogenase and NADH or NADPH can function as substrates for glycerol-3-phosphate dehydrogenase, but the complex with NADH is less active than free NADH, while the complex with NADPH is as active as free NADPH. Mg2+ enhanced the interactions between malate dehydrogenase and glycerol-3-phosphate dehydrogenase described above. Lactate dehydrogenase (E.C. 220.127.116.11) had effects similar to those of malate dehydrogenase only in the presence of Mg2+. In the absence of Mg2+, there was no evidence of interaction between lactate dehydrogenase and glycerol-3-phosphate dehydrogenase.


Effects of activation of sodium and calcium entry on spontaneous release of acetylcholine induced by methylmercury.
The effect of ionophores and channel activators for Ca and Na on the time course and magnitude of methylmercury (MeHg)-induced increase in spontaneous release of neurotransmitter was studied at the murine neuromuscular junction using intracellular microelectrode recording techniques. The goal was to test whether chemicals that increase entry of Na+ or Ca++ into nerve terminals would shorten the latent period that precedes the onset of MeHg-induced increase in MEPP frequency. Administration of MeHg (100 microM) with A23187 (25 microM), a calcium ionophore, caused a more rapid time to peak induced increase in MEPP frequency than "control" MeHg preparations. This effect also occurred in solutions to which no extracellular Ca++ was added. Use of monensin, a Na+ ionophore (25-100 microM), did not shorten the time to peak increase of MEPP frequency. The dihydropyridine Ca++ channel agonist Bay K 8644 (750 nM) produced the most marked shortening of the time to peak MEPP frequency for MeHg. This effect also occurred in solutions deficient in extracellular Ca++. Veratridine (20 microM), a sodium channel activator, decreased the time to peak MEPP frequency when used in conjunction with MeHg in both Ca++-containing and Ca++-deficient solutions. Replacement of sodium in the extracellular perfusion solution with methylamine, which does not penetrate axon sodium channels, did not prevent the MeHg-induced increase in MEPP frequency although it did prolong the time to peak increase and decreased the maximal MEPP frequency induced by MeHg compared with experiments conducted in sodium-containing solutions.(ABSTRACT TRUNCATED AT 250 WORDS)


Digital simulation of an arbitrary stationary stochastic process by spectral representation.
In this paper we present a straightforward, efficient, and computationally fast method for creating a large number of discrete samples with an arbitrary given probability density function and a specified spectral content. The method relies on initially transforming a white noise sample set of random Gaussian distributed numbers into a corresponding set with the desired spectral distribution, after which this colored Gaussian probability distribution is transformed via an inverse transform into the desired probability distribution. In contrast to previous work, where the analyses were limited to auto regressive and or iterative techniques to obtain satisfactory results, we find that a single application of the inverse transform method yields satisfactory results for a wide class of arbitrary probability distributions. Although a single application of the inverse transform technique does not conserve the power spectra exactly, it yields highly accurate numerical results for a wide range of probability distributions and target power spectra that are sufficient for system simulation purposes and can thus be regarded as an accurate engineering approximation, which can be used for wide range of practical applications. A sufficiency condition is presented regarding the range of parameter values where a single application of the inverse transform method yields satisfactory agreement between the simulated and target power spectra, and a series of examples relevant for the optics community are presented and discussed. Outside this parameter range the agreement gracefully degrades but does not distort in shape. Although we demonstrate the method here focusing on stationary random processes, we see no reason why the method could not be extended to simulate non-stationary random processes.


Associations of Blood Pressure Dipping Patterns With Left Ventricular Mass and Left Ventricular Hypertrophy in Blacks: The Jackson Heart Study.
Abnormal diurnal blood pressure (BP), including nondipping patterns, assessed using ambulatory BP monitoring, have been associated with increased cardiovascular risk among white and Asian adults. We examined the associations of BP dipping patterns (dipping, nondipping, and reverse dipping) with cardiovascular target organ damage (left ventricular mass index and left ventricular hypertrophy), among participants from the Jackson Heart Study, an exclusively black population-based cohort. Analyses included 1015 participants who completed ambulatory BP monitoring and had echocardiography data from the baseline visit. Participants were categorized based on the nighttime to daytime systolic BP ratio into 3 patterns: dipping pattern (≤0.90), nondipping pattern (>0.90 to ≤1.00), and reverse dipping pattern (>1.00). The prevalence of dipping, nondipping, and reverse dipping patterns was 33.6%, 48.2%, and 18.2%, respectively. In a fully adjusted model, which included antihypertensive medication use and clinic and daytime systolic BP, the mean differences in left ventricular mass index between reverse dipping pattern versus dipping pattern was 8.3±2.1 g/m2 (P<0.001) and between nondipping pattern versus dipping pattern was -1.0±1.6 g/m2 (P=0.536). Compared with participants with a dipping pattern, the prevalence ratio for having left ventricular hypertrophy was 1.65 (95% CI, 1.05-2.58) and 0.96 (95% CI, 0.63-1.97) for those with a reverse dipping pattern and nondipping pattern, respectively. In this population-based study of blacks, a reverse dipping pattern was associated with increased left ventricular mass index and a higher prevalence of left ventricular hypertrophy. Identification of a reverse dipping pattern on ambulatory BP monitoring may help identify black at increased risk for cardiovascular target organ damage.


Can Flu-Like Absenteeism in Kindergartens Be Reduced Through Hand Hygiene Training for Both Parents and Their Kindergarteners?
Background: Hand hygiene has been found as an effective method to prevent the spread of infectious diseases. Parents, however, were found to have inadequate knowledge, reporting skills, and parenting practices related to seasonal influenza. The aim of our study was to investigate if there was any correlation between the flu infection among parents and their kindergarteners and ascertain the effectiveness of using hand hygiene interventions in kindergartens to lower kindergarteners' absenteeism attributable to seasonal flu. Methods: This study was a quasi-experimental study with a pretest-posttest design. Fifteen kindergartens were randomly selected from the kindergarten lists in different districts displayed on the Hong Kong government website. From these 15 kindergartens, parents and their kindergarteners were then randomly selected to participate in our hand hygiene program. To support the validity of the program, the WHO hand hygiene checklist was used to ensure sufficient coverage of the objectives. All kindergarteners attended 45-minute session for 4 weeks, while their parents attended a separate session lasting 1 hour. Parents monitored their kindergartners on a daily basis for any flu symptoms and kindergartens monitored their school attendance. Results: The study results showed that kindergarteners with strong parenting and proper hand hygiene compliance had fewer recorded signs and symptoms of flu-like illnesses. Our findings also showed that the kindergarteners' absence rates in all participating kindergartens owing to flu decreased from 21.5% to 12% of the study period in 3 months. Conclusion: It was found that the flu infection rates of the parents and their kindergarteners were significantly correlated with P = .005. The awareness and personal hygiene skills of the parents and kindergarteners were both raised after the program. The findings in this study supported that positive parenting on hand hygiene can help reduce kindergarteners' flu-like absenteeism.


An examination of the potential added value of water safety plans to the United States national drinking water legislation.
National and sub-national governments develop and enforce regulations to ensure the delivery of safe drinking water in the United States (US) and countries worldwide. However, periodic contamination events, waterborne endemic illness and outbreaks of waterborne disease still occur, illustrating that delivery of safe drinking water is not guaranteed. In this study, we examined the potential added value of a preventive risk management approach, specifically, water safety plans (WSPs), in the US in order to improve drinking water quality. We undertook a comparative analysis between US drinking water regulations and WSP steps to analyze the similarities and differences between them, and identify how WSPs might complement drinking water regulations in the US. Findings show that US drinking water regulations and WSP steps were aligned in the areas of describing the water supply system and defining monitoring and controls. However, gaps exist between US drinking water regulations and WSPs in the areas of team procedures and training, internal risk assessment and prioritization, and management procedures and plans. The study contributes to understanding both required and voluntary drinking water management practices in the US and how implementing water safety plans could benefit water systems to improve drinking water quality and human health.


Vascular endothelial growth factor blockade rapidly elicits alternative proangiogenic pathways in neuroblastoma.
Most children with neuroblastoma presenting after infancy have metastatic, chemoresistant disease. Amplification of the MYCN proto-oncogene is a significant marker of these poor-prognosis neuroblastoma tumors. Recent studies suggest that MYCN may function in part by promoting angiogenesis via vascular endothelial growth factor (VEGF). VEGF blockade has been validated as a therapeutic strategy in adult cancers. In these studies, we asked whether inhibition of VEGF signaling via VEGFR2 blockade in established MYCN-amplified neuroblastoma xenografts would: 1) restrict tumor growth; 2) induce hypoxia; and 3) alter tumor vasculature. The MYCN-amplified neuroblastoma human cell line NGP was implanted intrarenally in athymic female mice. After 5 weeks, mice with established tumors were selected, a cohort euthanized to provide day 0 controls, and the rest assigned to receive biweekly injections of DC101 (anti-murine VEGFR2 antibody) or vehicle. DC101 treatment did not inhibit progressive tumor growth in established NGP xenografts. Although tumor vasculature was not significantly disrupted, a modest increase in tumor hypoxia was demonstrated by pimonidazole staining, and expression of a previously described hypoxia metagene was increased by gene set enrichment analysis (GSEA) in DC101-treated tumors. DC101 treatment elicited increased: 1) expression of VEGFR1 and its ligand placental growth factor; and 2) increased Notch activation in tumor vasculature concurrent with expression of the Notch ligand Jagged1. This result suggests that established MYCN-amplified neuroblastoma tumors are relatively VEGF-independent, and display the ability to rapidly up-regulate hypoxia-responsive alternative proangiogenic mechanisms that may stabilize vasculature when VEGF is deficient.


Metabolism of p-Cresol by the Fungus Aspergillus fumigatus.
The fungus Aspergillus fumigatus ATCC 28282 was shown to grow on p-cresol as its sole source of carbon and energy. A pathway for metabolism of this compound was proposed. This has protocatechuate as the ring-fission substrate with cleavage and metabolism by an ortho-fission pathway. The protocatechuate was formed by two alternative routes, either by initial attack on the methyl group, which is oxidized to carboxyl, followed by ring-hydroxylation, or by ring-hydroxylation as the first step with subsequent oxidation of 4-methylcatechol to the acid. The pathway was elucidated from several pieces of evidence. A number of compounds, including 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid, protocatechuic acid, protocatechualdehyde, and 4-methylcatechol, appeared transiently in the medium during growth on p-cresol. These compounds were oxidized without lag by p-cresol-grown cells but not by succinate-grown cells. Enzyme activities for most of the proposed steps were demonstrated in cell extracts after growth on p-cresol, and the products of these activities were identified. None of the activities were found in succinate-grown cells.


Molecular basis for coordinating transcription termination with noncoding RNA degradation.
The Nrd1-Nab3-Sen1 (NNS) complex is essential for controlling pervasive transcription and generating sn/snoRNAs in S. cerevisiae. The NNS complex terminates transcription of noncoding RNA genes and promotes exosome-dependent processing/degradation of the released transcripts. The Trf4-Air2-Mtr4 (TRAMP) complex polyadenylates NNS target RNAs and favors their degradation. NNS-dependent termination and degradation are coupled, but the mechanism underlying this coupling remains enigmatic. Here we provide structural and functional evidence demonstrating that the same domain of Nrd1p interacts with RNA polymerase II and Trf4p in a mutually exclusive manner, thus defining two alternative forms of the NNS complex, one involved in termination and the other in degradation. We show that the Nrd1-Trf4 interaction is required for optimal exosome activity in vivo and for the stimulation of polyadenylation of NNS targets by TRAMP in vitro. We propose that transcription termination and RNA degradation are coordinated by switching between two alternative partners of the NNS complex.


A comparison between subgaleal and subperiosteal brow lifts.
The author has thought for a long time that the endoscopic subperiosteal brow lift has a better long-term result than the bicoronal subgaleal brow lift; therefore, he stopped doing subgaleal bicoronal brow lifts in 1995 in favor of the endoscopic subperiosteal approach. To discover if this impression could be objectively verified, the author undertook a retrospective study of 120 patients who had a brow lift. They were divided into four groups, depending on the type of operation, and were compared with each other by means of a new digitalized analyzing tool. Measurements of the vertical height from the midpupil to the top of the brow, from the medial canthus to the top of the brow, and from the lateral canthus to the top of the brow were made. Preoperative measurements were compared with postoperative measurements taken after 1 year. No significant difference existed between the preoperative and postoperative values in the patients who had a subgaleal brow lift. In the patients who had a subperiosteal brow lift, there was a mean increase of 7 mm in the vertical height of the brow 1 year after the operation.


The effects of drugs inhibiting protein secretion in the filamentous fungus Trichoderma reesei. Evidence for down-regulation of genes that encode secreted proteins in the stressed cells.
To study the mechanisms of protein secretion as well as the cellular responses to impaired protein folding and transport in filamentous fungi, we have analyzed Trichoderma reesei cultures treated with chemical agents that interfere with these processes, dithiothreitol, brefeldin A, and the Ca(2+)-ionophore A23187. The effects of the drugs on the kinetics of protein synthesis and transport were characterized using metabolic labeling of synthesized proteins. Cellobiohydrolase I (CBHI, Cel7A), the major secreted cellulase, was analyzed as a model protein. Northern analysis showed that under conditions where protein transport was inhibited (treatments with dithiothreitol or brefeldin A) the unfolded protein response pathway was activated. The active form of the hac1 mRNA that mediates unfolded protein response signaling was induced, followed by induction of the foldase and chaperone genes pdi1 and bip1. Concomitant with the activation of the unfolded protein response pathway, the transcript levels of genes encoding secreted proteins, like cellulases and xylanases, were drastically decreased, suggesting a novel type of feedback mechanism activated in response to impairment in protein folding or transport (repression under secretion stress (RESS)). By studying expression of the reporter gene lacZ under cbh1 promoters of different length, it was shown that the feedback response was mediated through the cellulase promoter.


Risk factors and outcomes associated with post-traumatic headache after mild traumatic brain injury.
To determine the prevalence and potential risk factors of acute and chronic post-traumatic headache (PTH) in patients with mild to moderate traumatic brain injury (TBI) in a prospective longitudinal observational multicentre study. Acute PTH (aPTH) is defined by new or worsening of pre-existing headache occurring within 7 days after trauma, whereas chronic PTH (cPTH) is defined as persisting aPTH >3 months after trauma. An additional goal was to study the impact of aPTH and cPTH in terms of return to work (RTW), anxiety and depression. This was a prospective observational study conducted between January 2013 and February 2014 in three trauma centres in the Netherlands. Patients aged 16 years and older with a GCS score of 9-15 on admission to the ED, with loss of consciousness and/or amnesia were prospectively enrolled. Follow-up questionnaires were completed at 2 weeks and 3 months after injury with the Head Injury Symptom Checklist, the Hospital Anxiety and Depression Scale and RTW scale. In total, 628 patients were enrolled in the study, 469 completed the 2-week questionnaire (75%) at 2 weeks and 409 (65%) at 3 months. At 2 weeks, 238 (51%) had developed aPTH and at 3 months 95 (23%) had developed cPTH. Female gender, younger age, headache immediately at the ED and CT scan abnormalities increased the risk for aPTH. Risk factors for cPTH were female gender and headache at the ED. Patients with cPTH were less likely to have returned to work than those without cPTH (35% vs 14%, P=0.001). Patients with aPTH and cPTH more often report anxiety (20% and 28%, P=0.001) and depression (19% and 28%, P=0.001) after trauma in comparison with the group without PTH (10% anxiety and 8% depression). PTH is an important health problem with a significant impact on long-term outcome of TBI patients. Several risk factors were identified, which can aid in early identification of subjects at risk for PTH.


NF-κB/Rel Transcription Factors in Pancreatic Cancer: Focusing on RelA, c-Rel, and RelB.
Regulation of Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)/Rel transcription factors (TFs) is extremely cell-type-specific owing to their ability to act disparately in the context of cellular homeostasis driven by cellular fate and the microenvironment. This is also valid for tumor cells in which every single component shows heterogenic effects. Whereas many studies highlighted a per se oncogenic function for NF-κB/Rel TFs across cancers, recent advances in the field revealed their additional tumor-suppressive nature. Specifically, pancreatic ductal adenocarcinoma (PDAC), as one of the deadliest malignant diseases, shows aberrant canonical-noncanonical NF-κB signaling activity. Although decades of work suggest a prominent oncogenic activity of NF-κB signaling in PDAC, emerging evidence points to the opposite including anti-tumor effects. Considering the dual nature of NF-κB signaling and how it is closely linked to many other cancer related signaling pathways, it is essential to dissect the roles of individual Rel TFs in pancreatic carcinogenesis and tumor persistency and progression. Here, we discuss recent knowledge highlighting the role of Rel TFs RelA, RelB, and c-Rel in PDAC development and maintenance. Next to providing rationales for therapeutically harnessing Rel TF function in PDAC, we compile strategies currently in (pre-)clinical evaluation.


Family-centered service for children with cerebral palsy and their families: a review of the literature.
Pediatric neurologists and other specialists working with children with cerebral palsy or other disabilities play important roles in providing services in ways that make a difference in the lives of these children and their families. This article defines family-centered service and outlines a conceptual framework of the premises and principles underlying this approach to service delivery. Research evidence supporting family-centered service is presented for child, parent/family and service system outcomes, with a focus on community-based rehabilitation or health care services. The research evidence shows strong support for family-centered service in promoting the psychosocial well-being of children and their parents and in leading to increased satisfaction with services. There is less evidence for other outcomes. It is recommended that all service providers assess their beliefs and behaviors about family-centered service.


Overexpression of methyl-CpG binding protein 2 impairs T(H)1 responses.
The DNA binding protein methyl-CpG binding protein 2 (MeCP2) critically influences neuronal and brain function by modulating gene expression, and children with overexpression of the MECP2 gene exhibit postnatal neurological syndromes. We demonstrate that some children with MECP2 duplication also display variable immunological abnormalities that include reductions in memory T and B cells and natural killer cells and immunoglobulin assay responses. Moreover, whereas mice with MeCP2 overexpression were unable to control infection with the intra-macrophage parasite Leishmania major and secrete interferon-γ (IFN-γ) from involved lymph nodes, they were able to control airway fungal infection by Aspergillus niger and mount protective T helper cell type 2 (T(H)2)-dependent allergic responses. Relative to normal T cells, T(H) cells from children and mice with MECP2 duplication displayed similar impairments in IFN-γ secretion and T(H)1 responses that were due to both MeCP2-dependent suppression of IFN-γ transcription and sequestration of the IFN-γ locus as assessed by chromatin immunoprecipitation assay. Thus, overexpressed MeCP2 aberrantly suppresses IFN-γ secretion from T(H) cells, potentially leading to a partially immunodeficient state. Our findings establish a rational basis for identifying, treating, and preventing infectious complications potentially affecting children with MECP2 duplication.


Ex vivo gene therapy using intravitreal injection of GDNF-secreting mouse embryonic stem cells in a rat model of retinal degeneration.
Safe and prolonged drug delivery to the retina is a key obstacle to overcome in the development of new medicines aimed at treating progressive retinal disease. We took advantage of the ability of embryonic stem cells to survive long-term in foreign tissue and used these cells to deliver neuroprotectant molecules to the retina of the rhodopsin TgN S334ter-4 rat model of retinitis pigmentosa (RP). Mouse embryonic stem (mES) cells, derived from the pluripotent embryonic stem cell line E14TG2a, were genetically engineered to oversecrete the glial cell-derived neurotrophic factor (GDNF). Cell suspensions, containing approximately 200,000 cells and expressing approximately 35ng/10(6) cells/24 h GDNF, were injected into the vitreous cavity of TgN S334ter rat eyes at postnatal day 21 (P21) without immunosuppression. Histological and immunofluorescence imaging was used to evaluate photoreceptor survival up to P90. Local (vitreous) and systemic (serum) concentrations of GDNF were determined and ocular side effects were monitored. Green fluorescent protein (GFP)-expressing mES cells were observed on the inner limiting membrane of the retina in retinal flatmounts up to P90. In cryostat sections at P45, some GFP-expressing cells had integrated into the inner retina, but did not migrate into the outer nuclear layer. After an initial lag period, the photoreceptor cell counts were significantly higher (p< or =0.05) in animals treated with GDNF-secreting mES cells than in untreated animals, principally in the peripheral retina. Several adverse side effects such as tractional detachments and areas of hyperplasia were seen in a minimal number of treated eyes. Abnormally high levels of GDNF in the peripheral circulation were also observed. ES cells engineered to secrete GDNF exerted a neuroprotective effect for at least three months on retinal structure in the TgN S334ter rat model of retinal degeneration. Immunosuppression was not required for this. Several adverse effects were identified which require further investigation to make cell-based delivery of neuroprotection a viable clinical strategy.


Vascular pedicles of infrahyoid muscles: an anatomical study.
This study was designed to determine the entering sites of vascular pedicles of the infrahyoid muscles. The neck regions of 12 cadavers were investigated bilaterally. The vascular pedicles of the infrahyoid muscles (except the thyrohyoid and inferior belly of the omohyoid muscle) were dissected and measurements taken with a caliper in reference to the attachments of the muscles. For the sternothyroid muscle, two vascular pedicles were found bilaterally in all cadavers. Two vascular pedicles were found bilaterally for the sternohyoid muscle in 11 cadavers; in one cadaver the inferior vascular pedicle was absent bilaterally. One vascular pedicle was found bilaterally for the superior belly of the omohyoid muscle in all cadavers. These data regarding the vascular pedicles of the infrahyoid muscles might be useful for preserving the vascular supply of the infrahyoid muscles during flap preparation.


Stability and change in attachment security across adolescence.
This study examined both continuity and familial, intrapsychic, and environmental predictors of change in adolescent attachment security across a 2-year period from middle to late adolescence. Assessments included the Adult Attachment Interview, observed mother-adolescent interactions, test-based data, and adolescent self-reports obtained from an ethnically and socioeconomically diverse sample of moderately at-risk adolescents interviewed at ages 16 and 18. Substantial stability in security was identified. Beyond this stability, however, relative declines in attachment security were predicted by adolescents' enmeshed, overpersonalizing behavior with their mothers; depressive symptoms; and poverty status. Results suggest that although security may trend upward for nonstressed adolescents, stressors that overwhelm the capacity for affect regulation and that are not easily assuaged by parents predict relative declines in security over time.


Charged particle therapy for cancer: the inheritance of the Cavendish scientists?
The history of developments in atomic physics and its applications follows the decisive input provided by Maxwell and subsequent discoveries by his successors at the Cavendish Laboratory. In medicine the potential applications of particle physics (with the notable exception of the electron) were unfortunately delayed by the disappointing experiences with neutron therapy, which produced long-term scepticism. Neutrons are not appropriate for cancer therapy because not only their physical dose distributions offer no advantages over X-rays, but also their biological dose distributions are worse. The much improved dose distributions achieved with charged particles offer real prospects for better treatment outcomes because of the large reduction in the volume of unnecessarily irradiated tissue in many situations. Charged particle therapy is relatively new and can be applied with increasing confidence due to advances in radiology and computing, but at present there are insufficient numbers of treatment facilities to produce statistically powerful studies to compare treatment outcomes with those of X-rays. Considerable progress has been achieved in Japan and Germany with pilot studies of carbon ions but their efficacy compared with protons needs to be tested: in theory carbon should be better for intrinsically radio-resistant and for the most hypoxic tumours. The optimisation of proton and ion beam therapy in clinical practice remains to be achieved, but there are good scientific reasons why these modalities will be preferred by patients and their physicians in the future. Regrettably, despite hosting many of the momentous discoveries that enabled the development of charged particle therapy, the UK is slow to adopt and implement this very important form of cancer treatment.


The geographic variation of cancer incidence in Ontario.
Our objectives were (1) to describe an analysis of the spatial pattern of cancer incidence in Ontario and (2) to discuss the quality of data in the Ontario Cancer Registry with respect to the accuracy of local cancer rates. Cancer incidence rates were calculated for 22 cancer sites in 49 counties of Ontario during 1976 to 1986. Capture-recapture methods were used to estimate completeness of case registration, and completeness of residence information was also assessed. Spatial autocorrelation was used in measuring the geographic pattern of incidence rates. Comparisons were also made between sexes and with earlier data from 1966 to 1975. The quality of the geographic data in the registry appeared good, and corrections for incomplete or inaccurate registration had little impact. About one third of the sex-site combinations showed some evidence of spatial patterning in the cancer rate. Particularly strong regional variation was noted for cancers of the stomach, lung, uterus, and prostate. The analysis revealed a number of cancers with significant spatial patterning of risk. Further work is needed to relate the cancer data to other information on potential life-style and environmental factors.


Phototherapy in anti-aging and its photobiologic basics: a new approach to skin rejuvenation.
Intrinsic aging and photoaging of the face are constantly ongoing, and eventually result in the typical "aged" face, with visible lines and wrinkles at rest, a variety of dyschromia and a tired, dull and lax epidermis over poorly organized elastotic dermal architecture characterized by many interfibrillary spaces. Both ablative and nonablative resurfacing have been reported as solutions, the former providing excellent results, but a long patient downtime, and the latter giving little or no downtime, but less-than-ideal results. In ablative resurfacing, the epidermis is removed and replaced with a "new" epidermis, whereas in the nonablative approach the epidermis is spared through some form of cooling. In both approaches, however, the goal is to create controlled amounts of thermal damage in the dermis to stimulate the wound healing process, thus generating a tighter, better organized, "younger" dermal matrix. A better approach might be to apply prevention, rather than the cure, and to treat subjects in their very early 20s, before even fine lines have begun to appear. This "photoanti-aging" approach could be achieved with the use of very low incident levels of photon energy to stimulate the skin cells, both epidermal and dermal, at cell-specific wavelengths based on the photobiological findings of the literature over the past two decades or so, in order to increase their resistance to the effects of chronological and photoaging. Lasers and IPL systems could be used, but are extremely expensive and therapist-intensive. A new generation of light-emitting diodes (LEDs) has appeared as the result of a spin-off from the US NASA Space Medicine Program, which are much more powerful than the previous generation with quasimonochromatic outputs. These LEDs can offer target specificity to achieve photobiomodulated enhanced action potentials of the skin cells, in particular mast cells, macrophages, endotheliocytes, and fibroblasts, plus increases in local blood and lymphatic flow, in a noninvasive, athermal manner. New phototherapeutic LED-based systems have appeared to meet the need for a less-expensive but clinically useful light source to enable photoantiaging as a reality in clinical practice. Some studies proving the efficacy of LED therapy have already appeared, and based on their results LED therapy represents a potential new approach to prevention in anti-aging, so that further studies are warranted to prove its efficacy.


Interlaboratory performance of a microarray-based gene expression test to determine tissue of origin in poorly differentiated and undifferentiated cancers.
Clinical workup of metastatic malignancies of unknown origin is often arduous and expensive and is reported to be unsuccessful in 30 to 60% of cases. Accurate classification of uncertain primary cancers may improve with microarray-based gene expression testing. We evaluated the analytical performance characteristics of the Pathwork tissue of origin test, which uses expression signals from 1668 probe sets in a gene expression microarray, to quantify the similarity of tumor specimens to 15 known tissues of origin. Sixty archived tissue specimens from poorly and undifferentiated tumors (metastatic and primary) were analyzed at four laboratories representing a wide range of preanalytical conditions (eg, personnel, reagents, instrumentation, and protocols). Cross-laboratory comparisons showed highly reproducible results between laboratories, with correlation coefficients between 0.95 to 0.97 for measurements of similarity scores, and an average 93.8% overall concordance between laboratories in terms of final tissue calls. Bland-Altman plots (mean coefficients of reproducibility of 32.48+/-3.97) and kappa statistics (kappa >0.86) also indicated a high level of agreement between laboratories. We conclude that the Pathwork tissue of origin test is a robust assay that produces consistent results in diverse laboratory conditions reflecting the preanalytical variations found in the everyday clinical practice of molecular diagnostics laboratories.


A Rapid and Sensitive HPLC Method for Simultaneous Determination of Irinotecan Hydrochloride and Curcumin in Co-delivered Polymeric Nanoparticles.
In recent years, a great deal of attention has been paid to the combined use of multiple antitumor drugs for better cancer treatment. The aims of the study are to construct a nanoparticle drug delivery system for the co-delivery of irinotecan hydrochloride and curcumin and to develop an analytical method for simultaneously quantifying these molecules, which is essential for further studies of the co-delivered nano system. The irinotecan hydrochloride and curcumin co-delivered nanoparticle (ICN) were prepared by combinatorially entrapping them into polyethylene glycol-poly lactic acid-co-glycolic acid (PEG-PLGA) polymeric nanoparticles. A simple, sensitive and rapid high-performance liquid chromatography method was developed and validated to simultaneously quantify the compounds in the co-delivered nanoparticle system. Acetonitrile and ultrapure water containing sodium dodecyl sulfate (0.08 mol/L), disodium phosphate (Na2HPO4, 0.002 mol/L) and acetic acid (4%, v/v) were used as the mobile phase and their ratio was set at 50:50. The flow rate was set to 1.0 mL/min, and the temperature in the column oven was maintained at 40°C. The analysis was carried out at 256 and 424 nm to assess irinotecan hydrochloride and curcumin, respectively. Detectors with only one channel can also visualize both analytes in one chromatogram at 379 nm and still demonstrate acceptable sensitivity. The retention times for irinotecan hydrochloride and curium were 3.317 and 5.560 min, respectively. The method developed was confirmed to be sensitive, accurate (recovery, 100 ± 2%), precise (relative standard deviation, RSD ≤ 1%), robust and linear (R2 ≥ 0.9996) in the range from 2.05 to 1050 μg/mL. The presented method has been used to quantify irinotecan hydrochloride and curcumin in the co-delivered ICN nano system to assess the drug delivery quality of the nanoparticles and can also be used for routine analysis because of its simplicity and accuracy.


Structural analysis of thermostabilizing mutations of cocaine esterase.
Cocaine is considered to be the most addictive of all substances of abuse and mediates its effects by inhibiting monoamine transporters, primarily the dopamine transporters. There are currently no small molecules that can be used to combat its toxic and addictive properties, in part because of the difficulty of developing compounds that inhibit cocaine binding without having intrinsic effects on dopamine transport. Most of the effective cocaine inhibitors also display addictive properties. We have recently reported the use of cocaine esterase (CocE) to accelerate the removal of systemic cocaine and to prevent cocaine-induced lethality. However, wild-type CocE is relatively unstable at physiological temperatures (tau(1/2) approximately 13 min at 37 degrees C), presenting challenges for its development as a viable therapeutic agent. We applied computational approaches to predict mutations to stabilize CocE and showed that several of these have increased stability both in vitro and in vivo, with the most efficacious mutant (T172R/G173Q) extending half-life up to 370 min. Here we present novel X-ray crystallographic data on these mutants that provide a plausible model for the observed enhanced stability. We also more extensively characterize the previously reported variants and report on a new stabilizing mutant, L169K. The improved stability of these engineered CocE enzymes will have a profound influence on the use of this protein to combat cocaine-induced toxicity and addiction in humans.


Photoperiodic diapause under the control of circadian clock genes in an insect.
Most organisms have evolved a circadian clock in order to anticipate daily environmental changes and many of these organisms are also capable of sophisticated measurement of daylength (photoperiodism) that is used to regulate seasonal events such as diapause, migration and polymorphism. It has been generally accepted that the same elements are involved in both circadian (daily) and seasonal (annual) rhythms because both rely upon daily light-dark cycles. However, as reasonable as this sounds, there remains no conclusive evidence of such a molecular machinery in insects. We have approached this issue by using RNA interference (RNAi) in Riptortus pedestris. The cuticle deposition rhythm exhibited the major properties of circadian rhythms, indicating that the rhythm is regulated by a circadian clock. RNAi directed against the circadian clock genes of period and cycle, which are negative and positive regulators in the circadian clock, respectively, disrupted the cuticle deposition rhythm and distinct cuticle layers were produced by these RNAi. Simultaneously, period RNAi caused the insect to avert diapause under a diapause-inducing photoperiod whereas cycle RNAi induced diapause under a diapause-averting photoperiod. The expression patterns of juvenile hormone-regulated genes and the application of juvenile hormone analogue suggested that neither ovarian development itself nor a downstream cascade of juvenile hormone secretion, were disturbed by period and cycle RNAi. This study revealed that the circadian clock genes are crucial not only for daily rhythms but also for photoperiodic diapause. RNAi directed against period and cycle had opposite effects not only in the circadian cuticle deposition rhythm but also in the photoperiodic diapause. These RNAi also had opposite effects on juvenile hormone-regulated gene expression. It is still possible that the circadian clock genes pleiotropically affect ovarian development but, based on these results, we suggest that the circadian clock operated by the circadian clock genes, period and cycle, governs seasonal timing as well as the daily rhythms.


Factors Associated With Suicide Ideation in US Army Soldiers During Deployment in Afghanistan.
Understanding suicide ideation (SI) during combat deployment can inform prevention and treatment during and after deployment. To examine associations of sociodemographic characteristics, lifetime and past-year stressors, and mental disorders with 30-day SI among a representative sample of US Army soldiers deployed in Afghanistan. In this survey study, soldiers deployed to Afghanistan completed self-administered questionnaires in July 2012. The sample was weighted to represent all 87 032 soldiers serving in Afghanistan. Prevalence of lifetime, past-year, and 30-day SI and mental disorders was determined. Logistic regression analyses examined risk factors associated with SI. Data analyses for this study were conducted between August 2018 and August 2019. Suicide ideation, lifetime and 12-month stressors, and mental disorders were assessed with questionnaires. Administrative records identified sociodemographic characteristics and suicide attempts. A total of 3957 soldiers (3473 [weighted 87.5%] male; 2135 [weighted 52.6%] aged ≤29 years) completed self-administered questionnaires during their deployment in Afghanistan. Lifetime, past-year, and 30-day SI prevalence estimates were 11.7%, 3.0%, and 1.9%, respectively. Among soldiers with SI, 44.2% had major depressive disorder (MDD) and 19.3% had posttraumatic stress disorder in the past 30-day period. A series of analyses of the 23 grouped variables potentially associated with SI resulted in a final model of sex; race/ethnicity; lifetime noncombat trauma; past 12-month relationship problems, legal problems, and death or illness of a friend or family member; and MDD. In this final multivariable model, white race/ethnicity (odds ratio [OR], 3.1 [95% CI, 1.8-5.1]), lifetime noncombat trauma (OR, 2.1 [95% CI, 1.1-4.0]), and MDD (past 30 days: OR, 31.8 [95% CI, 15.0-67.7]; before past 30 days: OR, 4.9 [95% CI, 2.5-9.6]) were associated with SI. Among the 85 soldiers with past 30-day SI, from survey administration through 12 months after returning from deployment, 6% (5 participants) had a documented suicide attempt vs 0.14% (6 participants) of the 3872 soldiers without SI. This study suggests that major depressive disorder and noncombat trauma are important factors in identifying SI risk during combat deployment.


Role of multiple microRNAs in the sexually dimorphic expression of Cyp2b9 in mouse liver.
Mouse cytochrome P450 2b9 (Cyp2b9) is a testosterone 16α-hydroxylase enzyme showing female-specific expression in many inbred mouse strains, including C57BL/6J. Previous studies have recognized that some sex-dependently secreted endogenous modulating factors were involved in the sexually dimorphic expression of Cyp2b9 through transcriptional regulation. In this study, we found evidence that some microRNAs contributed to the sexually biased expression of Cyp2b9 via post-transcriptional regulation. Cyp2b9 was upregulated in livers of hepatocyte-specific Dicer1 knockout mice at 3 weeks. The age-dependent downregulation of Cyp2b9 in the livers of male mice was diminished when Dicer1 was specifically knocked out in hepatocytes. When these data were combined with bioinformatics analysis and microRNA profiles of male and female mice, we found that 18 microRNAs were associated with the sexually dimorphic expression of Cyp2b9, which showed higher expression levels in male C57BL/6J mice when compared with females. Luciferase assays revealed that approximate half of these microRNAs repressed luciferase activity in a reporter system containing the 3'-untranslated region (3'-UTR) of Cyp2b9, and also inhibited Cyp2b9 protein expression. MicroRNA seed region mutation or mutations in putative binding sites of the microRNAs in Cyp2b9 3'-UTR led to the loss of the suppression of luciferase activity. There was also a negative correlation between the levels of these microRNAs and Cyp2b9. Our results suggested that multiple microRNAs participated in the regulation of Cyp2b9 expression, and that the lower expression levels of these microRNAs potentially contributed to the female-specific expression of Cyp2b9 in the livers of C57BL/6J mice.


Percutaneous Coronary Intervention in Native Coronary Arteries Versus Bypass Grafts in Patients With Prior Coronary Artery Bypass Graft Surgery: Insights From the Veterans Affairs Clinical Assessment, Reporting, and Tracking Program.
The aim of this study was to examine the frequency, associations, and outcomes of native coronary artery versus bypass graft percutaneous coronary intervention (PCI) in patients with prior coronary artery bypass grafting (CABG) in the Veterans Affairs (VA) integrated health care system. Patients with prior CABG surgery often undergo PCI, but the association between PCI target vessel and short- and long-term outcomes has received limited study. A national cohort of 11,118 veterans with prior CABG who underwent PCI between October 2005 and September 2013 at 67 VA hospitals was examined, and the outcomes of patients who underwent native coronary versus bypass graft PCI were compared. Logistic regression with generalized estimating equations was used to adjust for correlation between patients within hospitals. Cox regressions were modeled for each outcome to determine the variables with significant hazard ratios (HRs). During the study period, patients with prior CABG represented 18.5% of all patients undergoing PCI (11,118 of 60,171). The PCI target vessel was a native coronary artery in 73.4% and a bypass graft in 26.6%: 25.0% in a saphenous vein graft and 1.5% in an arterial graft. Compared with patients undergoing native coronary artery PCI, those undergoing bypass graft PCI had higher risk characteristics and more procedure-related complications. During a median follow-up period of 3.11 years, bypass graft PCI was associated with significantly higher mortality (adjusted HR: 1.30; 95% confidence interval: 1.18 to 1.42), myocardial infarction (adjusted HR: 1.61; 95% confidence interval: 1.43 to 1.82), and repeat revascularization (adjusted HR: 1.60; 95% confidence interval: 1.50 to 1.71). In a national cohort of veterans, almost three-quarters of PCIs performed in patients with prior CABG involved native coronary artery lesions. Compared with native coronary PCI, bypass graft PCI was significantly associated with higher incidence of short- and long-term major adverse events, including more than double the rate of in-hospital mortality.


Skin autoreactivity in Hashimoto's thyroiditis patients without urticaria: autologous serum skin test positivity correlation with thyroid antibodies, sonographical volume and grading.
Recent studies have shown an association between anti-thyroid antibodies and autologous serum skin test (ASST) positive urticaria patients. However, a connection between thyroid and this reliable skin test for mast cell autoreactivity, ASST, has not been reported yet. We investigated ASST in patients with Hashimoto's thyroiditis (HT) without urticaria and compared the results with laboratory and sonographical findings of HT. 154 HT patients, 100 healthy volunteers without HT as a first control group and 46 patients with multinodular goitre but without autoimmune thyroid disease as a second control group underwent testing with ASST. ASST was applied to these groups according to two criteria, first as ASST(new): autologous serum red wheal response 1.5 mm bigger than negative control; second as ASST(old): serum red wheal response 5 mm bigger than negative control accepted as positive. Free triiodothyronine (fT3), free thyroxine (fT4), thyroid-stimulating hormone (TSH), thyroid peroxidase antibody (anti-TPO) and thyroglobulin antibody (anti-Tg) levels were measured. ASST(old), ASST(new) scored positive in 51.3-60.4% of HT patients, with statistically significant differences. Thyroid volume grades were inversely proportional with ASST(old) and (new) positivity. Moderate (+) titers of anti-Tg in ASST(old) and (new) (+) cases were significantly higher than the same titers of anti-Tg in ASST(old) and (new) (-) cases. The prevalence of ASST positivity in HT patients was not affected by the following factors: gender, age at screening, laboratory measurements of thyroid function tests, anti-TPO antibodies and thyroid ultrasound (US) echogenicity. Positivity of ASST in HT has shown that there is a skin mast cell autoreactivity in HT patients independent of autoreactive chronic urticaria (ACU).


Pneumococcal vaccines for preventing pneumonia in chronic obstructive pulmonary disease.
People with chronic obstructive pulmonary disease (COPD) are at increased risk of pneumococcal disease, especially pneumonia, as well as acute exacerbations with associated morbidity and healthcare costs. To determine the efficacy of injectable pneumococcal vaccination for preventing pneumonia in persons with COPD. We searched the Cochrane Airways COPD Trials Register and the databases CENTRAL, MEDLINE and Embase, using prespecified terms. Searches are current to November 2016. We included randomised controlled trials (RCT) comparing injectable pneumococcal polysaccharide vaccine (PPV) or pneumococcal conjugated vaccine (PCV) versus a control or alternative vaccine type in people with COPD. We used standard Cochrane methodological procedures. For meta-analyses, we subgrouped studies by vaccine type. For this update, we added five studies (606 participants), meaning that the review now includes a total of 12 RCTs involving 2171 participants with COPD. Average age of participants was 66 years, male participants accounted for 67% and mean forced expiratory volume in one second (FEV1) was 1.2 L (five studies), 54% predicted (four studies). We assessed risks of selection, attrition and reporting bias as low, and risks of performance and detection bias as moderate.Compared with control, the vaccine group had a lower likelihood of developing community-acquired pneumonia (CAP) (odds ratio (OR) 0.62, 95% confidence interval (CI) 0.43 to 0.89; six studies, n = 1372; GRADE: moderate), but findings did not differ specifically for pneumococcal pneumonia (Peto OR 0.26, 95% CI 0.05 to 1.31; three studies, n = 1158; GRADE: low). The number needed to treat for an additional beneficial outcome (NNTB) (preventing one episode of CAP) was 21 (95% CI 15 to 74). Mortality from cardiorespiratory causes did not differ between vaccine and control groups (OR 1.07, 95% CI 0.69 to 1.66; three studies, n = 888; GRADE: moderate), nor did all-cause mortality differ (OR 1.00, 95% CI 0.72 to 1.40; five studies, n = 1053; GRADE: moderate). The likelihood of hospital admission for any cause, or for cardiorespiratory causes, did not differ between vaccine and control groups. Vaccination significantly reduced the likelihood of a COPD exacerbation (OR 0.60, 95% CI 0.39 to 0.93; four studies, n = 446; GRADE: moderate). The NNTB to prevent a patient from experiencing an acute exacerbation was 8 (95% CI 5 to 58). Only one study (n = 181) compared the efficacy of different vaccine types - 23-valent PPV versus 7-valent PCV - and reported no differences for CAP, all-cause mortality, hospital admission or likelihood of a COPD exacerbation, but investigators described a greater likelihood of some mild adverse effects of vaccination with PPV-23. Injectable polyvalent pneumococcal vaccination provides significant protection against community-acquired pneumonia, although no evidence indicates that vaccination reduced the risk of confirmed pneumococcal pneumonia, which was a relatively rare event. Vaccination reduced the likelihood of a COPD exacerbation, and moderate-quality evidence suggests the benefits of pneumococcal vaccination in people with COPD. Evidence was insufficient for comparison of different pneumococcal vaccine types.


Sister chromatid cohesion control and aneuploidy.
Apart from a personal tragedy, could Down syndrome, cancer and infertility possibly have something in common? Are there links between a syndrome with physical and mental problems, a tumor growing out of control and the incapability to reproduce? These questions can be answered if we look at the biological functions of a protein complex, named cohesin, which is the main protagonist in the regulation of sister chromatid cohesion during chromosome segregation in cell division. The establishment, maintenance and removal of sister chromatid cohesion is one of the most fascinating and dangerous processes in the life of a cell. Errors in the control of sister chromatid cohesion frequently lead to cell death or aneuploidy. Recent results showed that cohesins also have important functions in non-dividing cells, revealing new, unexplored roles for these proteins in human syndromes, currently known as cohesinopathies. In the last 10 years, we have improved our understanding of the molecular mechanisms of the cohesin and cohesin-interacting proteins regulating the different events of sister chromatid cohesion during cell division in mitosis and meiosis.


Parasitic modulation of host development by ubiquitin-independent protein degradation.
Certain obligate parasites induce complex and substantial phenotypic changes in their hosts in ways that favor their transmission to other trophic levels. However, the mechanisms underlying these changes remain largely unknown. Here we demonstrate how SAP05 protein effectors from insect-vectored plant pathogenic phytoplasmas take control of several plant developmental processes. These effectors simultaneously prolong the host lifespan and induce witches' broom-like proliferations of leaf and sterile shoots, organs colonized by phytoplasmas and vectors. SAP05 acts by mediating the concurrent degradation of SPL and GATA developmental regulators via a process that relies on hijacking the plant ubiquitin receptor RPN10 independent of substrate ubiquitination. RPN10 is highly conserved among eukaryotes, but SAP05 does not bind insect vector RPN10. A two-amino-acid substitution within plant RPN10 generates a functional variant that is resistant to SAP05 activities. Therefore, one effector protein enables obligate parasitic phytoplasmas to induce a plethora of developmental phenotypes in their hosts.


Problem-solving abilities and frontal lobe cortical thickness in healthy aging and mild cognitive impairment.
Mild cognitive impairment (MCI) is considered a transitional state between normal aging and Alzheimer disease. Most MCI subjects present disturbances in multiple neuropsychological domains, including executive function. This study aimed at exploring frontal lobe cortical thinning in MCI and healthy controls, and its relationship with problem-solving abilities. Twenty-three MCI patients and 30 elderly controls underwent MRI and neuropsychological assessment. Cortical thickness was measured by means of FreeSurfer. Problem-solving was assessed by means of the Tower of London (TOL) task. MCI showed a global thinning of the cortex. With regard to specific regions of interest, a thinning in the left frontal lobe and the bilateral posterior cingulate gyri was found. Partial correlations, after controlling for age, education, Mini-Mental Status Examination, and non-frontal mean thickness revealed negative significant correlations between frontal lobe thickness and executive outcomes in the control group. This counterintuitive relationship was not observed in the MCI group, suggesting that the frontal cortical atrophy observed in MCI entails a specific pathology-related relationship with high-level executive outcomes that is qualitatively different from that observed in healthy aging.


AACR centennial series: the biology of cancer metastasis: historical perspective.
Metastasis resistant to therapy is the major cause of death from cancer. Despite almost 200 years of study, the process of tumor metastasis remains controversial. Stephen Paget initially identified the role of host-tumor interactions on the basis of a review of autopsy records. His "seed and soil" hypothesis was substantiated a century later with experimental studies, and numerous reports have confirmed these seminal observations. An improved understanding of the metastatic process and the attributes of the cells selected by this process is critical for the treatment of patients with systemic disease. In many patients, metastasis has occurred by the time of diagnosis, so metastasis prevention may not be relevant. Treating systemic disease and identifying patients with early disease should be our goal. Revitalized research in the past three decades has focused on new discoveries in the biology of metastasis. Even though our understanding of molecular events that regulate metastasis has improved, the contributions and timing of molecular lesion(s) involved in metastasis pathogenesis remain unclear. Review of the history of pioneering observations and discussion of current controversies should increase understanding of the complex and multifactorial interactions between the host and selected tumor cells that contribute to fatal metastasis and should lead to the design of successful therapy.


Lifestyle modifications to prevent and control hypertension. 7. Recommendations on stress management. Canadian Hypertension Society, Canadian Coalition for High Blood Pressure Prevention and Control, Laboratory Centre for Disease Control at Health Canada, Heart and Stroke Foundation of Canada.
To provide updated evidence-based recommendations for health care professionals concerning the effects of stress management on the prevention and control of hypertension in otherwise healthy adults (except pregnant women). Alternatives to stress management include other nonpharmacologic interventions and medical therapy; these options are not mutually exclusive. The health outcome considered was reduction of blood pressure. There is little evidence to date that stress management prevents death or vascular events. Because of insufficient evidence, no economic outcomes were considered. A systematic search of the literature (which yielded, among other sources, 3 meta-analyses) was conducted for the period 1966-1997 with the terms essential hypertension, treatment, psychological, behavioural, cognitive, relaxation, mediation, biofeedback and stress management. Other relevant evidence was obtained from the reference lists of the articles identified, from the personal files of the authors and through contacts with experts. The articles were reviewed, classified according to study design and graded according to level of evidence. A high value was placed on the avoidance of cardiovascular morbidity and premature death caused by uncontrolled hypertension. The magnitude of the reduction in blood pressure obtained with multicomponent, individualized cognitive behavioural intervention for stress management was comparable in some studies to that obtained with weight loss or drugs; single-component interventions such as biofeedback or relaxation were less effective. The adverse effects of stress-management techniques are minimal, but the cost for effective interventions is substantial, similar initially to drug costs; continuing costs are probably minimal. (1) In patients with hypertension, the contribution of stress should be considered. (2) For hypertensive patients in whom stress appears to be an important issue, stress management should be considered as an intervention. Individualized cognitive behavioural interventions are more likely to be effective than single-component interventions. These recommendations were reviewed by all of the sponsoring organizations and by participants in a satellite symposium of the fourth International Conference on Preventive Cardiology. They have not been clinically tested. The Canadian Hypertension Society, the Canadian Coalition for High Blood Pressure Prevention and Control, the Laboratory Centre for Disease Control at Health Canada, and the Heart and Stroke Foundation of Canada.


Optical Coherence Tomography Investigations and Modeling of the Sintering of Ceramic Crowns.
Dental prostheses are sintered in ovens that sometimes suffer from a loss of calibration. This can lead to variations of the sintering temperature outside the range recommended by the manufacturer. Stress and even fractures in dental ceramics may occur, and this leads to the necessity to rebuild the dental construct. The aim of this work is to monitor the quality of sintering processes using an established biomedical imaging technique-optical coherence tomography (OCT). Conventional current procedures imply the fabrication of supplemental samples that add to the expenses and are only evaluated visually. To our knowledge, we were the first to propose the use of OCT, a non-destructive method that brings objectivity for such assessments, focusing, in a previous study, on metal ceramic dental prostheses. Here, a different material, pressed ceramics, is considered, while we propose a quantitative assessment of the results-using reflectivity profiles of en-face (i.e., constant-depth) OCT images of sintered samples. The results for both the pressed ceramics and metal ceramics prostheses are discussed by obtaining the analytic functions of their reflectivity profiles. A multi-parametric analysis demonstrates the best parameter to characterize the loss of calibration of dental ovens. Rules-of-thumb are extracted; producing dental prostheses with defects can thus be avoided.


Manganese and iron deficiency in Southern Ocean Phaeocystis antarctica populations revealed through taxon-specific protein indicators.
Iron and light are recognized as limiting factors controlling Southern Ocean phytoplankton growth. Recent field-based evidence suggests, however, that manganese availability may also play a role. Here we examine the influence of iron and manganese on protein expression and physiology in Phaeocystis antarctica, a key Antarctic primary producer. We provide taxon-specific proteomic evidence to show that in-situ Southern Ocean Phaeocystis populations regularly experience stress due to combined low manganese and iron availability. In culture, combined low iron and manganese induce large-scale changes in the Phaeocystis proteome and result in reorganization of the photosynthetic apparatus. Natural Phaeocystis populations produce protein signatures indicating late-season manganese and iron stress, consistent with concurrently observed stimulation of chlorophyll production upon additions of manganese or iron. These results implicate manganese as an important driver of Southern Ocean productivity and demonstrate the utility of peptide mass spectrometry for identifying drivers of incomplete macronutrient consumption.


Basal and Postprandial Myofibrillar Protein Synthesis Rates Do Not Differ between Lean and Obese Middle-Aged Men.
Excess lipid availability has been associated with the development of anabolic resistance. As such, obesity may be accompanied by impairments in muscle protein metabolism. We hypothesized that basal and postprandial muscle protein synthesis rates are lower in obese than in lean men. Twelve obese men [mean ± SEM age: 48 ± 2 y; BMI (in kg/m2): 37.0 ± 1.5; body fat: 32 ± 2%] and 12 age-matched lean controls (age: 43 ± 3 y; BMI: 23.4 ± 0.4; body fat: 21 ± 1%) received primed continuous L-[ring-2H5]-phenylalanine and L-[ring-3,5-2H2]-tyrosine infusions and ingested 25 g intrinsically L-[1-13C]-phenylalanine labeled whey protein. Repeated blood and muscle samples were obtained to assess protein digestion and amino acid absorption kinetics, and basal and postprandial myofibrillar protein synthesis rates. Exogenous phenylalanine appearance rates increased after protein ingestion in both groups (P < 0.001), with a total of 53 ± 1% and 53 ± 2% of dietary protein-derived phenylalanine appearing in the circulation over the 5-h postprandial period in lean and obese men, respectively (P = 0.82). After protein ingestion, whole-body protein synthesis and oxidation rates increased to a greater extent in lean men than in the obese (P-interaction < 0.05), resulting in a higher whole-body protein net balance in the lean than in the obese (7.1 ± 0.2 and 4.6 ± 0.4 µmol phenylalanine · h-1 · kg-1, respectively; P-interaction < 0.001). Myofibrillar protein synthesis rates increased from 0.030 ± 0.002 and 0.028 ± 0.003%/h in the postabsorptive period to 0.034 ± 0.002 and 0.035 ± 0.003%.h-1 in the 5-h postprandial period (P = 0.03) in lean and obese men, respectively, with no differences between groups (P-interaction = 0.58). Basal, postabsorptive myofibrillar protein synthesis rates do not differ between lean and obese middle-aged men. Postprandial protein handling, including protein digestion and amino acid absorption, and the postprandial muscle protein synthetic response after the ingestion of 25 g whey protein are not impaired in obese men. This trial was registered at www.trialregister.nl as NTR4060.


The Effect of a Pex3 Mutation on Hearing and Lipid Content of the Inner Ear.
Peroxisome biogenesis disorders (due to PEX gene mutations) are associated with symptoms that range in severity and can lead to early childhood death, but a common feature is hearing impairment. In this study, mice carrying Pex3 mutations were found to show normal auditory development followed by an early-onset progressive increase in auditory response thresholds. The only structural defect detected in the cochlea at four weeks old was the disruption of synapses below inner hair cells. A conditional approach was used to establish that Pex3 expression is required locally within the cochlea for normal hearing, rather than hearing loss being due to systemic effects. A lipidomics analysis of the inner ear revealed a local reduction in plasmalogens in the Pex3 mouse mutants, comparable to the systemic plasmalogen reduction reported in human peroxisome biogenesis disorders. Thus, mice with Pex3 mutations may be a useful tool to understand the physiological basis of peroxisome biogenesis disorders.


Effect of early infant feeding practices on infection-specific neonatal mortality: an investigation of the causal links with observational data from rural Ghana.
Strong associations between delayed initiation of breastfeeding and increased neonatal mortality (2-28 d) were recently reported in rural Ghana. Investigation into the biological plausibility of this relation and potential causal pathways is needed. The objective was to assess the effect of early infant feeding practices (delayed initiation, prelacteal feeding, established neonatal breastfeeding) on infection-specific neonatal mortality in breastfed neonates aged 2-28 d. This prospective observational cohort study was based on 10 942 breastfed singleton neonates born between 1 July 2003 and 30 June 2004, who survived to day 2, and whose mothers were visited in the neonatal period. Verbal autopsies were used to ascertain the cause of death. One hundred forty neonates died from day 2 to day 28; 93 died of infection and 47 of noninfectious causes. The risk of death as a result of infection increased with increasing delay in initiation of breastfeeding from 1 h to day 7; overall late initiation (after day 1) was associated with a 2.6-fold risk [adjusted odds ratio (adj OR): 2.61; 95% CI: 1.68, 4.04]. Partial breastfeeding was associated with a 5.7-fold adjusted risk of death as a result of infectious disease (adj OR: 5.73; 95% CI: 2.75, 11.91). No obvious associations were observed between these feeding practices and noninfection-specific mortality. Prelacteal feeding was not associated with infection (adj OR: 1.11; 95% CI: 0.66, 1.86) or noninfection-specific (adj OR: 1.33; 95% CI: 0.55, 3.22) mortality. This study provides the first epidemiologic evidence of a causal association between early breastfeeding and reduced infection-specific neonatal mortality in young human infants.


Changing Developmental Trajectories of Toddlers With Autism Spectrum Disorder: Strategies for Bridging Research to Community Practice.
The need for community-viable, evidence-based intervention strategies for toddlers with autism spectrum disorder (ASD) is a national priority. The purpose of this research forum article is to identify gaps in intervention research and needs in community practice for toddlers with ASD, incorporate published findings from a randomized controlled trial (RCT) of the Early Social Interaction (ESI) model (Wetherby et al., 2014) to illustrate community-based intervention, report new findings on child active engagement from the ESI RCT, and offer solutions to bridge the research-to-community practice gap. Research findings were reviewed to identify gaps in the evidence base for toddlers with ASD. Published and new findings from the multisite ESI RCT compared the effects of two different ESI conditions for 82 toddlers with ASD to teach parents how to support active engagement in natural environments. The RCT of the ESI model was the only parent-implemented intervention that reported differential treatment effects on standardized measures of child outcomes, including social communication, developmental level, and adaptive behavior. A new measure of active engagement in the natural environment was found to be sensitive to change in 3 months for young toddlers with ASD and to predict outcomes on the standardized measures of child outcomes. Strategies for utilizing the Autism Navigator collection of web-based courses and tools using extensive video footage for families and professional development are offered for scaling up in community settings to change developmental trajectories of toddlers with ASD. Current health care and education systems are challenged to provide intervention of adequate intensity for toddlers with ASD. The use of innovative technology can increase acceleration of access to evidence-based early intervention for toddlers with ASD that addresses health disparities, enables immediate response as soon as ASD is suspected, and rapidly bridges the research-to-practice gap.


Effect of practice on standardised learning outcomes in simulation-based medical education.
This report synthesises a subset of 31 journal articles on high-fidelity simulation-based medical education containing 32 research studies drawn from a larger qualitative review published previously. These studies were selected because they present adequate data to allow for quantitative synthesis. We hypothesised an association between hours of practice in simulation-based medical education and standardised learning outcomes measured as weighted effect sizes. Journal articles were screened using 5 exclusion and inclusion criteria. Response data were extracted and 3 judges independently coded each study. Learning outcomes were standardised using a common metric, the average weighted effect size (AWES), due to the heterogeneity of response measures in individual studies. anova was used to evaluate AWES differences due to hours of practice on a high-fidelity medical simulator cast in 5 categories. The eta squared (eta2) statistic was used to assess the association between AWES and simulator practice hours. There is a strong association (eta2=0.46) between hours of practice on high-fidelity medical simulators and standardised learning outcomes. The association approximates a dose-response relationship. Hours of high-fidelity simulator practice have a positive, functional relationship with standardised learning outcomes in medical education. More rigorous research methods and more stringent journal editorial policies are needed to advance this field of medical education research.


Herbalists of Today's Switzerland and Their Plant Knowledge. A Preliminary Analysis from an Ethnobotanical Perspective.
Herbal medicine is a popular part of primary health care in Switzerland. Following an ethnobotanical approach, in this study we seek to identify Swiss herbalists with broad, empirical medicinal plant knowledge and use. We aim to consider different areas of the medicinal landscape including biomedicine, complementary and alternative medicine, and self-medication. A total of 61 expert interviews were conducted from February 2010 to November 2011. The transfer of knowledge was analyzed according to a Switzerland-related selection of historical and recent popular as well as scientific herbal books. A total of 254 medicinal plant species, belonging to 218 genera and 87 families, were recorded in 934 use reports. Predominantly leaves and flowers are used for the treatment of dermatological, respiratory, nervous, and gastrointestinal problems. Recent to historical herbal books are an important source of plant knowledge. Medicinal plants are used for self-medication and professional health care and despite different underlying medicinal concepts and philosophies, herbalists largely agree on the most important medicinal plant species.


Anticoagulant repertoire of the hookworm Ancylostoma caninum.
Hookworms are hematophagous nematodes that infect a wide range of mammalian hosts, including humans. There has been speculation for nearly a century as to the identity of the anticoagulant substances) used by these organisms to subvert host hemostasis. Using molecular cloning, we describe a family of potent small protein (75-84 amino acids) anticoagulants from the hookworm Ancylostoma caninum termed AcAP (A. caninum anticoagulant protein). Two recombinant AcAP members (AcAP5 and AcAP6) directly inhibited the catalytic activity of blood coagulation factor Xa (fXa), while a third form (AcAPc2) predominantly inhibited the catalytic activity of a complex composed of blood coagulation factor VIIa and tissue factor (fVIIa/TF). The inhibition of fVIIa/TF was by a unique mechanism that required the initial formation of a binary complex of the inhibitor with fXa at a site on the enzyme that is distinct from the catalytic center (exo-site). The sequence of AcAPc2 as well as the utilization of an exo-site on fXa distinguishes this inhibitor from the mammalian anticoagulant TFPI (tissue factor pathway inhibitor), which is functionally equivalent with respect to fXa-dependent inhibition of fIIa/TF. The relative sequence positions of the reactive site residues determined for AcAP5 with the homologous regions in AcAP6 and AcAPc2 as well as the pattern of 10 cysteine residues present in each of the inhibitors suggest that the AcAPs are distantly related to the family of small protein serine protease inhibitors found in the nonhematophagous nematode Ascaris lumbricoides var. suum.


Significance of a fragmented QRS complex in patients with chronic total occlusion of coronary artery without prior myocardial infarction.
Fragmented QRS (fQRS) complexes that have numerous RSR´ patterns represent alteration of ventricular depolarization. We evaluated the relationship between fQRS and poor coronary collateral circulation and the diagnostic ability of fQRS for myocardial scar detection in patients with chronic total occlusion (CTO) without a history of myocardial infarction. The study population consisted of patients undergoing coronary angiography with a suspicion of CAD. Seventy-nine patients with one totally occluded major coronary artery were enrolled. Exclusion criteria were history of MI; recent acute coronary syndrome; pathologic Q wave on 12-lead ECG; cardiomyopathy or severe valvular disease; coronary artery bypass surgery or percutaneous coronary angioplasty. Collateral circulation was scored on the basis of Rentrop's classification. All patients were assessed by myocardial perfusion SPECT. Fragmented QRS was characterized as existence of an R´ or R wave or S wave notch in two adjacent leads related to the location of a major coronary artery region. Single and multiple logistic regression analyses were completed in the forward method. Forty-nine patients had poor and 30 had well-developed collateral circulation. Fragmented QRS complexes were significantly higher in the poor collateral group (81% vs. 20%, p<0.001). Sensitivity, specificity, and the positive and negative predictive values of fQRS for myocardial scar identification were 89.4%, 87.5%, and 91.3% and 84.8%, respectively. The summed stress score and the summed rest score on SPECT were significantly higher in the poor collateral group than in the well-developed group (p<0.001) as well as in the fQRS group than the non-fQRS group (p<0.001). Logistic regression analysis revealed that the presence of fQRS was significantly and independently associated with poor collateral circulation and myocardial scar in patients with CTO. Fragmented QRS is independently related to poor coronary collateral circulation in patients with CTO without prior myocardial infarction. Notably, it can be a good predictor of myocardial scar rather than merely ischemia, with high diagnostic accuracy.