An improved flow cytometry-based natural killer cytotoxicity assay involving calcein AM staining of effector cells.
Several flow cytometric methods for measuring natural killer cell activity have been developed. Commonly used protocols involve the staining of target cells with various fluorescent dyes. However, these protocols are not applicable to certain experimental settings. Therefore, we used Calcein AM (CAM), which has been reported to be the most suitable dye for use in target cell staining protocols, as a means of developing an improved flow cytometry-based NK cytotoxicity assay involving effector cell staining. Peripheral blood mononuclear cells (PBMCs) isolated by gradient density centrifugation and expanded NK cells were used as effector cells. Cytotoxicity against K562 cells and several hematologic cancer cell lines was measured by a flow cytometry-based method using CAM and propidium iodide. The new assay was compared with a standard (51)Cr release assay (CRA) in terms of its ability to measure the cytotoxicity of NK cells in PBMCs and expanded NK cells against K562 cells. The optimal concentration of CAM for staining effector cells was 0.05 μM, and CAM fluorescence intensity in effector cells was maintained for 4 hours. CAM staining had no significant effect on NK cell activity in human PBMCs or expanded NK cells. Comparison of the CRA and this new assay using K562 cells revealed a good correlation (PBMCs, r = 0.894; expanded NK cells, r = 0.887). Distinct separation between target tumor cells (Daudi, Raji, RPMI8226, U266, U937, and K562 cells) and CAM-stained PBMCs (E:T ratio, 12.5:1 to 50:1) or expanded NK cells (E:T ratio, 0.5 to 4:1) was observed after incubation for 1 or 4 hours. In summary, we successfully developed an effective flow cytometry-based assay for assessing the activity of NK cells in PBMCs and expanded NK cells against K562 cells and various types of hematologic cancer cells.


BDNF-induced changes in the expression of the translation machinery in hippocampal neurons: protein levels and dendritic mRNA.
BDNF plays a key role in neuronal development, in short- and long-term changes in synaptic activity, and in neuronal survival. These effects are mediated, to a great extent, by changes in protein synthesis. We conducted a gel-based proteome profiling of the long-term (12 h) effects of BDNF in cultured hippocampal neurons. BDNF changed the abundance of proteins involved in (i) Nucleobase, nucleoside, nucleotide and nucleic acid metabolism, (ii) protein metabolism, (iii) carbohydrate metabolism, (iv) regulators of apoptosis, and (v) regulators of cell proliferation. A large majority of the identified proteins involved in translation activity were upregulated, but not all changes in the protein content were correlated with alterations in the corresponding mRNA. The upregulation of Seryl-aminoacyl-tRNA-synthetase and Eef2 was sensitive to the mTOR inhibitor rapamycin, as determined by Western blot. Since the mRNAs for proteins involved in translation represent a large fraction of the diversity of dendritic mRNAs, we investigated the effect of BDNF on the distribution of the transcripts in the soma versus neurite compartments. The increase in mRNA for proteins of the translation machinery in the soma was differentially coupled with the upregulation of neurite transcripts. BDNF also downregulated specific mRNAs in neurite compartments suggesting that the neurotrophin may act by regulating mRNA stability and thereby affecting the dendritic protein content.


Succinoglycan is required for initiation and elongation of infection threads during nodulation of alfalfa by Rhizobium meliloti.
Rhizobium meliloti Rm1021 must be able to synthesize succinoglycan in order to invade successfully the nodules which it elicits on alfalfa and to establish an effective nitrogen-fixing symbiosis. Using R. meliloti cells that express green fluorescent protein (GFP), we have examined the nature of the symbiotic deficiency of exo mutants that are defective or altered in succinoglycan production. Our observations indicate that an exoY mutant, which does not produce succinoglycan, is symbiotically defective because it cannot initiate the formation of infection threads. An exoZ mutant, which produces succinoglycan without the acetyl modification, forms nitrogen-fixing nodules on plants, but it exhibits a reduced efficiency in the initiation and elongation of infection threads. An exoH mutant, which produces symbiotically nonfunctional high-molecular-weight succinoglycan that lacks the succinyl modification, cannot form extended infection threads. Infection threads initiate at a reduced rate and then abort before they reach the base of the root hairs. Overproduction of succinoglycan by the exoS96::Tn5 mutant does not reduce the efficiency of infection thread initiation and elongation, but it does significantly reduce the ability of this mutant to colonize the curled root hairs, which is the first step of the invasion process. The exoR95::Tn5 mutant, which overproduces succinoglycan to an even greater extent than the exoS96::Tn5 mutant, has completely lost its ability to colonize the curled root hairs. These new observations lead us to propose that succinoglycan is required for both the initiation and elongation of infection threads during nodule invasion and that excess production of succinoglycan interferes with the ability of the rhizobia to colonize curled root hairs.


Risk of serious adverse events associated with biologic and nonbiologic psoriasis systemic therapy: patients ineligible vs eligible for randomized controlled trials.
To describe the use of systemic therapy for psoriasis (biologic and nonbiologic [classic] drugs) in patients not adequately represented in randomized controlled trials (RCTs) and the risk of serious adverse events (SAEs) in these patients. A registry inception cohort was used. Thirteen dermatology departments in Spain participated. A consecutive sample of patients treated with biologics and a systematic sample of patients treated with classic systemic therapy were evaluated. A total of 1042 patients (2179 person-years) were included. Inadequate representation in trials was defined as the presence of any of the following factors: elderly age (>70 years); type of psoriasis other than chronic plaque psoriasis; history of infection caused by hepatitis B, hepatitis C, or human immunodeficiency virus; history of cancer (excluding nonmelanoma skin cancer); and chronic renal or hepatic disease. Serious adverse events as defined by the International Conference on Harmonization were evaluated. In all, 29.8% of patients receiving systemic therapy for psoriasis would not have been eligible for RCTs. These individuals had an increased risk of SAEs (incidence rate ratio, 2.7; 95% CI, 1.5-4.7). Patients exposed to biologics had an adjusted increased risk of SAEs (incidence rate ratio, 2.3; 95% CI, 1.1-4.8) that was similar in patients eligible and ineligible for RCTs. Patients ineligible for RCTs are an important proportion (30%) of those receiving systemic therapy for psoriasis. These patients have a higher risk of SAEs and should be closely monitored. Patients exposed to biologics (whether these patients are eligible for RCTs or ineligible) are susceptible to the same increase in risk of SAEs, but biologics add to a higher baseline risk in patients who are ineligible for RCTs. The risk-benefit ratio in ineligible patients receiving biologics might be different from the ratio in eligible patients.


Reflex responses to combined hip and knee motion in human chronic spinal cord injury.
The relative contributions of hip and knee proprioceptors to the origination of extensor spasms were examined in 11 subjects with chronic spinal cord injury (SCI). Ramp and hold extension and combined hip and knee oscillation movements were imposed to the right leg while the ankle was held in a static position by a custom-designed robot. Isometric joint torques of the hip, knee, and ankle and surface electromyograms (EMGs) from seven leg muscles were recorded following controlled hip and knee extension. A stereotypical torque response consisting of hip flexion, knee extension, and ankle plantar flexion was observed following hip and knee perturbations. Further, the hip or knee joint posture modulated the spastic reflexes triggered by the extension movement of the other joint, with larger responses observed with the hip and knee extended. In addition, combined hip and knee oscillation movements were imposed to one leg with four different phasing conditions. The phasing between the hip and knee modulated the reflex activity triggered by hip and knee oscillations. The EMG patterns of the spastic reflexes were generally consistent with muscle timing during locomotion in human SCI. This knowledge may help identify rehabilitation strategies that produce functional movements in human SCI.


Information complementarity: A new paradigm for decoding quantum incompatibility.
The existence of observables that are incompatible or not jointly measurable is a characteristic feature of quantum mechanics, which lies at the root of a number of nonclassical phenomena, such as uncertainty relations, wave--particle dual behavior, Bell-inequality violation, and contextuality. However, no intuitive criterion is available for determining the compatibility of even two (generalized) observables, despite the overarching importance of this problem and intensive efforts of many researchers. Here we introduce an information theoretic paradigm together with an intuitive geometric picture for decoding incompatible observables, starting from two simple ideas: Every observable can only provide limited information and information is monotonic under data processing. By virtue of quantum estimation theory, we introduce a family of universal criteria for detecting incompatible observables and a natural measure of incompatibility, which are applicable to arbitrary number of arbitrary observables. Based on this framework, we derive a family of universal measurement uncertainty relations, provide a simple information theoretic explanation of quantitative wave--particle duality, and offer new perspectives for understanding Bell nonlocality, contextuality, and quantum precision limit.


Hemolytic anemia in wild seaducks caused by marine oil pollution.
Clinico-pathological examinations were conducted on wild white-winged scoters (Melanitta fusca) contaminated with fuel oil (Bunker C oil) from a capsized cargo ship in February 1993 in Japan. The erythrocyte count, hemoglobin concentration and hematocrit value in the oiled seaducks all were decreased and numerous immature erythrocytes were observed in blood smears. In addition, hemosiderosis was observed in the liver, kidney, and lung of some birds. We propose that the sea-ducks suffered from hemolytic anemia induced by ingestion of oil, which occurs when the birds preen their oiled plumage.


The effects of severe functional mitral regurgitation on right ventricular function in patients with advanced heart failure who were on waiting list for heart transplant.
This study aims to investigate the effects of severe functional mitral regurgitation on the parameters that reflect right ventricular function such as tricuspid annular plane systolic excursion and right ventricular stroke work index in potential heart transplant recipients. Between January 2015 and January 2017, a total of 282 consecutive patients (250 males, 32 females; mean age: 46±10 years; range, 18 to 66 years) with advanced heart failure who were referred for heart transplantation were retrospectively analyzed. The patients were divided into two groups as severe (n=84) and non-severe functional mitral regurgitation (n=198). Patients" medical histories, demographic characteristics, echocardiographic evaluations, and findings of right heart catheterization were recorded. The two groups were similar in terms of left ventricular ejection fraction, the New York Heart Association functional class, Interagency Registry for Mechanically Assisted Circulatory Support profile, and the duration of heart failure (p>0.05). Both groups were also similar with respect to tricuspid annular plane systolic excursion and right ventricular stroke work index. Functional mitral regurgitation was the only statistically significant variable in the univariate analysis for tricuspid annular plane systolic excursion (odds ratio [OR]: 0.58; 95% confidence interval [CI] 0.34-0.97; p=0.04), with no significant effect in the multivariate analysis. In the univariate analysis for right ventricular stroke work index, pulmonary arterial systolic pressure (OR: 0.77; 95% CI 0.67-0.88; p<0.001) was a significant variable and also had a significant effect in the multivariate analysis (OR: 0.92; 95% CI 0.87-0.97; p=0.003). In the tertile analyses, there were no significant differences between the two groups with respect to tricuspid annular plane systolic excursion and right ventricular stroke work index. We found no significant difference in right ventricular functions between the severe and non-severe functional mitral regurgitation groups in patients with advanced heart failure who had relatively short follow-up. Right ventricle can maintain its normal function at early stage. Adaptive remodeling of right ventricle may have an effect on these findings. Severe functional mitral regurgitation may be associated with adverse effects on advanced heart failure by increasing the right ventricular afterload.


Economic issues relating to access to medications.
Millions of uninsured, low-income workers (a disproportionate number of them black) are financially unable to obtain prescription drugs for treatment of conditions such as hypertension. As a result, funds spent to diagnose their underlying illnesses may be wasted, and they are likely to suffer complications that require much more costly care. The tendency for income levels to be inversely correlated with blood pressure means that those least able to pay for antihypertensive medication are those most susceptible to complications. Since current programs are inadequate to help most of these patients, there is a need for a joint initiative by the federal government and the pharmaceutical industry to fund programs to make medications easily accessible for the medically indigent population.


International comparison of contributions to psychosocial research on survivors of childhood cancer: past and future considerations.
To compare the methods and designs, constructs, publication rate, and location of studies in the past 15 years of pediatric psycho-oncology research, across geographic areas and over time. Three recently published reviews on psychological consequences of surviving childhood cancer were reanalyzed according to (1) inclusion of young or older survivors, (2) 5-year periods of publication, (3) country/area (North America, Europe, or Australia), (4) journal type, (5) number of survivors, (6) design of the study, (7) inclusion of a control group, (8) diagnosis, and (9) the use of standardized instruments on different domains. The comparisons show some descriptive differences depending on the origin of study, as well as difference over time. Over the years an increase in research from Europe/Australia over that from North America is shown. In most studies more constructs are measured during the more recent publication period. If we are to conduct exemplary work in this area, it is vital to establish national and international organizations for psychologists to parallel those already established for physicians. More attention should be focused on development of interventions for survivors in the future.


Sex and spouse conditions influence symptoms of anxiety, depression, and posttraumatic stress disorder in both patients admitted to intensive care units and their spouses.
To assess the effect of sex and spouse condition on symptoms of anxiety, depression and posttraumatic stress symptoms in patients and their spouses. A prospective study conducted in a 22-bed adult mixed intensive care unit in a tertiary hospital in São Paulo, Brazil. Patients and their spouses were enrolled 2 days after intensive care unit admission. They were interviewed while in the intensive care unit using the Hospital Anxiety and Depression Scale. At 30 and 90 days after intensive care unit discharge, they completed the Impact of Event Scale and Hospital Anxiety and Depression Scale by phone. From March 2011 to March 2013, we analyzed 118 patients and their spouses. We observed that female sex was associated with higher scores than male sex was in terms of the anxiety Hospital Anxiety and Depression Scale - subscale (p = 0.032) and depression (p = 0.034). There was no association between sex and posttraumatic stress disorder symptoms. However, spouses had higher Impact of Event Scale points compared with patients (p = 0.001). Female sex was associated with anxiety and depression, and spouses were more vulnerable to post-traumatic stress symptoms than the patients were. Increasing age and a later time of assessment were also associated with lower scores on the Impact of Event Scale.


The cell biology of lignification in higher plants.
Lignin is a polyphenolic polymer that strengthens and waterproofs the cell wall of specialized plant cell types. Lignification is part of the normal differentiation programme and functioning of specific cell types, but can also be triggered as a response to various biotic and abiotic stresses in cells that would not otherwise be lignifying. Cell wall lignification exhibits specific characteristics depending on the cell type being considered. These characteristics include the timing of lignification during cell differentiation, the palette of associated enzymes and substrates, the sub-cellular deposition sites, the monomeric composition and the cellular autonomy for lignin monomer production. This review provides an overview of the current understanding of lignin biosynthesis and polymerization at the cell biology level. The lignification process ranges from full autonomy to complete co-operation depending on the cell type. The different roles of lignin for the function of each specific plant cell type are clearly illustrated by the multiple phenotypic defects exhibited by knock-out mutants in lignin synthesis, which may explain why no general mechanism for lignification has yet been defined. The range of phenotypic effects observed include altered xylem sap transport, loss of mechanical support, reduced seed protection and dispersion, and/or increased pest and disease susceptibility.


A (five-)level playing field for mental health conditions?: exploratory analysis of EQ-5D-5L-derived utility values.
Economic evaluations of mental health interventions often measure health benefit in terms of utility values derived from the EQ-5D. For the five-level version of the EQ-5D, there are two methods of estimating utility [crosswalk and stated preference (5L-SP)]. This paper explores potential impacts for researchers and decision-makers when comparing utility values derived from either method in the specific context of mental health. Baseline EQ-5D-5L data from three large randomised controlled trials of interventions for mental health conditions were analysed. Utility values were generated using each method. Mean utility values were compared using a series of t tests on pooled data and subgroups. Scenario analyses explored potential impacts on cost-effectiveness decisions. EQ-5D data were available for 1399 participants. The mean utility value for each trial was approximately 0.08 higher when estimated using the 5L-SP approach compared to crosswalk (p < 0.0001). The difference was greatest among people reporting extreme anxiety/depression (mean utility 5L-SP 0.309, crosswalk 0.084; difference = 0.225; p < 0.0001). Identical improvements in health status were associated with higher costs to gain one QALY with the 5L-SP approach; this is more pronounced when improvements are across all domains compared to improvements on the anxiety/depression domain only. The two approaches produce significantly different utility values in people with mental health conditions. Resulting differences in cost per QALY estimates suggest that thresholds of cost-effectiveness may also need to be reviewed. Researchers and decision-makers should exercise caution when comparing or synthesising data from trials of mental health interventions using different utility estimation approaches.


Clinical use of homograft stored by vitrification.
Using 20% DMSO and 6% propylene glycol in Kreb' Ringer phosphate solution as cryopotective agent for homograft vitrification storage, the viability of stored homograft (79.2%) was higher than that of slow cooling storage (59.7%). About 540000 cm2 of vitrified homograft were used to cover the wounds after excision of burn eschar in 135 patients with major burns. The take-rate was over 94%. The cryopreservation of skin by vitrification can improve the viability and quality of skin and save time and cost.


The quorum-quenching lactonase from Geobacillus caldoxylosilyticus: purification, characterization, crystallization and crystallographic analysis.
Lactonases are enzymes that are capable of hydrolyzing various lactones such as aliphatic lactones or acyl-homoserine lactones (AHLs), with the latter being used as chemical signaling molecules by numerous Gram-negative bacteria. Lactonases therefore have the ability to quench the chemical communication, also known as quorum sensing, of numerous bacteria, and in particular to inhibit behaviors that are regulated by this system, such as the expression of virulence factors or the production of biofilms. A novel representative from the metallo-β-lactamase superfamily, dubbed GcL, was isolated from the thermophilic bacterium Geobacillus caldoxylosilyticus. Because of its thermophilic origin, GcL may constitute an interesting candidate for the development of biocontrol agents. Here, we show that GcL is a thermostable enzyme with a half-life at 75°C of 152.5 ± 10 min. Remarkably, it is also shown that GcL is among the most active lactonases characterized to date, with catalytic efficiencies (kcat/Km) against AHLs of greater than 10(6) M(-1) s(-1). The structure of GcL is expected to shed light on the catalytic mechanism of the enzyme and the molecular determinants for the substrate specificity in this class of lactonases. Here, the expression, purification, characterization, crystallization and X-ray diffraction data collection to 1.6 Å resolution of GcL are reported.


[Some data of the lipid changes disturbing of blood, heart and lungs after lead exposition].
Lipid changes after lead exposition were investigated. The experiment was made on "Wistar" line full-grown rats. Diving two months the experimental animals were given acetic acid lead diluted with water--evaluated in kilogram weight pure lead 0,3 (I group) and 1,9 (II group) milligrams during the rained. Low-level lead exposition causes disturbing the lipid changes of the erythrocytes, heart muscle and lungs. The changes are in the correlation with the loading intensity the body with the lead--as more lead ions are used in the experiment, as deeper is the destruction changes.


Adapting the "Chester step test" to predict peak oxygen uptake in children.
Maximal exercise testing may be difficult to perform in clinical practice, especially in obese children who have low cardiorespiratory fitness and exercise tolerance. We aimed to elaborate a model predicting peak oxygen consumption (VO2) in lean and obese children with use of the submaximal Chester step test. We performed a maximal step test, which consisted of 2-minute stages with increasing intensity to exhaustion, in 169 lean and obese children (age range: 7-16 years). VO2 was measured with indirect calorimetry. A statistical Tobit model was used to predict VO2 from age, gender, body mass index (BMI) z-score and intensity levels. Estimated VO2peak was then determined from the heart rate-VO2 linear relationship extrapolated to maximal heart rate (220 minus age, in beats.min-1). VO2 (ml/kg/min) can be predicted using the following equation: VO2 = 22.82 - [0.68*BMI z-score] - [0.46*age (years)] - [0.93*gender (male = 0; female = 1)] + [4.07*intensity level (stage 1, 2, 3 etc.)] - [0.24*BMI z-score *intensity level] - [0.34*gender*intensity level]. VO2 was lower in participants with high BMI z-scores and in female subjects. The Chester step test can assess cardiorespiratory fitness in lean and obese children in clinical settings. Our adapted equation allows the Chester step test to be used to estimate peak aerobic capacity in children.


Randomised trial comparing buprenorphine and diamorphine for chest pain in suspected myocardial infarction.
Buprenorphine, a new powerful analgesic agent, was used to treat chest pain in patients with suspected myocardial infarction. Initial studies showed no significant changes in systemic or pulmonary artery blood pressure or in heart rate after intravenous buprenorphine. Sublingual buprenorphine also appeared effective in relieving pain, but its onset of action was considerably delayed compared with the intravenous route. A randomised double-blind controlled trial of equivalent doses of buprenorphine and diamorphine showed no significant difference between the drugs in terms of pain relief and duration of action. The occurrence of nausea, vomiting, and other side effects was similar in the two groups. The onset of action of buprenorphine was slightly but significantly slower than that of diamorphine. Since buprenorphine seems to be comparable with diamorphine in action and is not a controlled drug, it may prove useful in both general and hospital practice.


Telephone-Delivered Dietary Intervention in Patients with Age-Related Macular Degeneration: 3-Month Post-Intervention Findings of a Randomised Controlled Trial.
There is an evidence-practice gap between the dietary recommendations for age-related macular degeneration (AMD) presented in the literature and those practiced by patients. This study reports on the 3-month post-intervention results of a randomised controlled trial (RCT) evaluating telephone-delivered counselling to improve dietary behaviours among AMD patients. A total of 155 AMD patients (57% female, aged 78 ± 8 years; control: 78, intervention: 77), primarily residing in New South Wales, Australia, were recruited. Participants completed a baseline questionnaire and a short dietary questionnaire (SDQ-AMD). The intervention included an evidence-based nutrition resource and four monthly calls with a dietitian. Immediately post-intervention, intervention participants repeated the SDQ-AMD and completed a feedback form. At 3 months post-intervention, both study arms repeated the SDQ-AMD. Statistical analyses included t-tests and McNemar's test. Intervention participants reported satisfaction with the tailored phone calls, nutrition resource and nutrition education provided. At 3 months post-intervention, there was no statistically significant difference between study arms in the proportion of participants meeting the dietary goals nor in intake (mean servings ± SE) of total vegetables (primary outcome) and other key food groups; however, there was a significantly higher intake of nuts (secondary outcome) (3.96 ± 0.51 vs. 2.71 ± 0.32; p = 0.04) among participants in the intervention versus control group. Within the intervention arm, there were also significant improvements in intakes of the following secondary outcomes: dark green leafy vegetables (0.99 ± 0.17 vs. 1.71 ± 0.22; p = 0.003) and legumes (0.69 ± 0.10 vs. 1.12 ± 0.16; p = 0.02) and intake of sweets and processed/prepared foods (8.31 ± 0.76 vs. 6.54 ± 0.58, p = 0.01). In summary, although there were few dietary differences between study arms at 3 months post-intervention, the intervention involving four monthly calls was acceptable and helpful to the participants. This type of intervention therefore has the potential to provide people with AMD the needed support for improving their nutrition knowledge and dietary practices, especially if continued over a longer period.


Investigating the locus of transposed-phoneme effects using cross-modal priming.
In this study, we examined whether the facilitatory priming effect found when auditory primes and targets are related by a phoneme transposition (e.g., /ʀͻb/-/bͻʀ/: Dufour & Grainger, 2019, 2020) is also observed under cross-modal presentation. In two experiments using the same materials as in the previous studies, we found no evidence for a facilitatory priming effect when the targets were presented visually rather than auditorily. On the contrary, an inhibitory priming effect was found when both unrelated words (Experiment 1; e.g., /mas/-/bͻR/) and vowel overlap words (Experiment 2; e.g., /vͻl/-/bͻʀ/) were used as control conditions. In Experiment 2, this inhibitory effect was found to be equivalent in size whether the target words were of higher or lower frequency than the prime words (e.g. /ʀͻb/-/bͻʀ/ vs. /bͻʀ/-/ʀͻb/). We interpret this pattern of effects as reflecting the greater impact of word-level inhibition in cross-modal priming, and the parallel influence of prime-target relative frequency on bottom-up phoneme-to-word facilitation and word-level inhibition. Therefore, the facilitatory priming effect previously observed with auditory primes and targets would mainly reflect bottom-up activation of the target word representation during prime word processing.


Prescribers' Knowledge, Attitudes and Behaviors on Antibiotics, Antibiotic Use and Antibiotic Resistance in Jordan.
More research is needed on the drivers of irrational antibiotic prescribing among healthcare professionals and to ensure effective prescribing and an adequate understanding of the issue of antibiotic resistance. This study aimed at evaluating prescribers' knowledge, attitudes and behaviors about antibiotic use and antibiotic resistance. A cross-sectional study was conducted utilizing an online questionnaire and included physicians and dentists from all sectors in Jordan. A total of 613 prescribers were included (physicians n = 409, dentists n = 204). Respondents' knowledge on effective use, unnecessary use or associated side effects of antibiotics was high (>90%), compared with their knowledge on the spread of antibiotic resistance (62.2%). For ease of access to the required guidelines on managing infections, and to materials that advise on prudent antibiotic use and antibiotic resistance, prescribers agreed in 62% and 46.1% of cases, respectively. 28.4% of respondents had prescribed antibiotics when they would have preferred not to do so more than once a day or more than once a week. Among respondents who prescribed antibiotics, 63.4% would never or rarely give out resources on prudent use of antibiotics for infections. The findings are of importance to inform antibiotic stewardships about relevant interventions aimed at changing prescribers' behaviors and improving antibiotic prescribing practices.


Metalloenzyme-Mediated Thiol-Yne Addition Towards Photoisomerizable Fluorescent Dyes.
Proteins are able to irreversibly assemble biologically active ligands from building blocks bearing complementary reactive functions due their spatial proximity, through a kinetic target-guided synthetic process (also named in situ click chemistry). Although linkages thus formed are mostly passive, some of them have shown to significantly contribute to the protein binding through for instance hydrogen bonding and stacking interactions. Biocompatible reactions and click chemistry are a formidable source of inspiration for developing such new protein-directed ligations. This study reports a proximity-induced thiol-yne synthesis of carbonic anhydrase inhibitors. Not only this example widens the arsenal of kinetic target-guided synthesis (KTGS) eligible reactions, but the obtained product displayed unsuspected photophysical properties. The corresponding vinyl sulfide linkage conjugated to a coumarin core proved to be engaged in a monodirectional Z to E photoisomerization process. Further investigations guided by theoretical calculations showed that fine-tuning of the nature of the substituents on the coumarin moiety allows to obtain a bidirectional photochemical process, thus discovering a new photoswitching moiety, displaying moreover fluorescence properties. Due to the spectral tunability of coumarin derivatives, this work should open new opportunities for the design of vinyl sulfide-based photoswitch systems with modular photophysical properties.


Statistical analysis of diffusion coefficient determination by fluorescence correlation spectroscopy.
Fluorescence correlation spectroscopy (FCS) has become an important and widely used technique for many applications in physics, chemistry, and biology. The parameter most frequently addressed by FCS is the diffusion of molecules in solution. Due to the highly non-linear connection between the diffusion coefficient and a measured autocorrelation function, it is extremely difficult to analyse the accuracy of the diffusion-coefficient determination in a FCS experiment. Here, we present a simplified analysis based on some general maximum-likelihood considerations, and numerical result are given for the dependence of the accuracy of the diffusion-coefficient determination on sample concentration, brightness, and measurement time. Optimal concentration values for performing FCS are found.


Safe Distance of Virus Quantitative Analysis and Simulation of the Trajectory of Pathogen-Containing Droplets in the Air Respiratory Airways.
Droplets of mucosalivary ejecta emitted by sneezing or coughing are a major carrier of numerous types of bacterial and viral diseases. This study develops a numerical model to estimate the spread distance for inhalable droplets (1-50 μm) in the air, the inhalability of the particles, and the trajectory as well as velocity of these pathogen-containing droplets in human respiratory airways. Moreover, particularly for droplets with diameters of 1 μm, 5 μm, 10 μm, and 50 μm, specific comparisons between their inhalability and transmission velocities are made. Data extracted from previous experiments proceeded by other researchers discussing the visualization of sneeze ejecta and deposition features of inhaled drops were used to obtain parameters to fit the model prediction of this work. Currently, research on similar topics was mostly based on either experiments or theoretical calculations only on one specific clan of pathogen, while the novel contribution of this paper is the combination and comparison of these two distinct methodologies that can be applied to solve a general practical problem aiming to all types of viruses by considering the pathogen-containing droplets as a whole entity.


Assessment of Sport Nutrition Knowledge, Dietary Practices, and Sources of Nutrition Information in NCAA Division III Collegiate Athletes.
Nutrition knowledge is a critical component of meeting sport nutrition guidelines. The present study aimed to evaluate the sport nutrition knowledge of National Collegiate Athletic Association (NCAA) Division III (DIII) athletes using a validated questionnaire, and to assess the dietary practices and sources of nutrition information in this population. A total of 331 student-athletes (n = 149 males, n = 181 females, n = 1 no sex indicated) completed the questionnaire. The mean score for total sport nutrition knowledge was 6.49 ± 8.9 (range -49 to 49) with a mean percent (%) correct score of 36.9 ± 19.1%. Athletes who had a previous college-level nutrition course (n = 62) had significantly higher (p < 0.05) total sport nutrition, carbohydrate, and hydration knowledge compared to those who did not (n = 268). Individual sport athletes (n = 90) scored significantly higher (p < 0.05) on hydration and micronutrients knowledge than team sport athletes (n = 237), while females scored higher than males for hydration knowledge (p < 0.05). The majority of athletes reported sensible dietary habits, such as not frequently skipping meals and eating carbohydrate and protein foods peri-workout. Athletes also reported their primary sources of nutrition information, the top three sources being social media, coaches, and athletic trainers, despite most frequently rating registered dietitians/nutritionists as "extremely knowledgeable". Despite low sport nutrition knowledge, NCAA DIII collegiate athletes practiced seemingly prudent dietary habits but lacked exposure to high-quality sources of nutrition information.


Contribution of small and medium enterprises to economic development and quality of life in Turkey.
All successful cities are the most developed and productive cities in their regions and countries in terms of economy and quality of life. In addition, it is observed that there are strong industries and enterprises in these regions and cities. Therefore, this study tries to evaluate the quality of life and the development of the industry together. The proposed method can be considered as a combination of operational research and GIS (Geographical Information System). The results of both approaches support each other. In addition, the innovative approaches of the sectors are evaluated by TOPSIS (Technique for Order Preference by Similarity to Ideal Solution). Also, the article recommends SMEs (Small and Medium-Sized Enterprises) for sustainable economic growth. According to the results, the quality of life and socio-economic development has a very close relationship with the existence of investments and manufacturing companies. If the current development policy continues, the regions with low quality of life will continue to lag behind the other cities in terms of development. The study indicates that there are remarkable differences between eastern and western Turkey in terms of living standards and the contribution of SMEs to economic development.


Search for an association between the human CYP1A2 genotype and CYP1A2 metabolic phenotype.
The genotype responsible for more than 60-fold interindividual differences in human hepatic CYP1A2 constitutive expression is not understood. Resequencing the human CYP1A1_CYP1A2 locus (39.6 kb) in five major geographically isolated subgroups recently led to the identification of 85 single nucleotide polymorphisms (SNPs), 57 of which were double-hit SNPs. Here, we attempted to correlate the CYP1A2 genotype with a metabolic phenotype. We chose 16 SNPs (all having a minor allele frequency > or =0.05 in Caucasians) to genotype 32 DNA samples (26 Caucasians, six Ethiopians) in which CYP1A2 metabolism had previously been determined. From 280 subjects (five locations worldwide) that had been CYP1A2-phenotyped, we genotyped the 10 highest, 14 lowest and eight intermediate DNA samples. Although no SNP was significant (P<0.05), possibly due to the small sample size, we found a trend for several of the six SNPs across the CYP1A2 linkage disequilibrium block associated with the trait. Five CYP1A2 haplotypes were inferred, two of which had not previously been reported; haplotype 1A2H10 showed the greatest association with CYP1A2 activity. Regulatory sequences responsible for the large interindividual differences in hepatic CYP1A2 gene basal expression might reside, in part, with some of these CYP1A2 SNPS but, in large part, might be located either cis (in nearby sequences not yet haplotyped) or trans in that they are not linked to the gene. We conclude that no SNP or haplotype in the CYP1A2 gene has yet been identified that can unequivocally be used to predict the metabolic phenotype in any individual patient.


Antiaging effects of dietary supplements and natural products.
Aging is an inevitable process influenced by genetics, lifestyles, and environments. With the rapid social and economic development in recent decades, the proportion of the elderly has increased rapidly worldwide, and many aging-related diseases have shown an upward trend, including nervous system diseases, cardiovascular diseases, metabolic diseases, and cancer. The rising burden of aging-related diseases has become an urgent global health challenge and requires immediate attention and solutions. Natural products have been used for a long time to treat various human diseases. The primary cellular pathways that mediate the longevity-extending effects of natural products involve nutrient-sensing pathways. Among them, the sirtuin, AMP-activated protein kinase, mammalian target of rapamycin, p53, and insulin/insulin-like growth factor-1 signaling pathways are most widely studied. Several studies have reviewed the effects of individual natural compounds on aging and aging-related diseases along with the underlying mechanisms. Natural products from food sources, such as polyphenols, saponins, alkaloids, and polysaccharides, are classified as antiaging compounds that promote health and prolong life via various mechanisms. In this article, we have reviewed several recently identified natural products with potential antiaging properties and have highlighted their cellular and molecular mechanisms. The discovery and use of dietary supplements and natural products that can prevent and treat multiple aging-related diseases in humans will be beneficial. Thus, this review provides theoretical background for existing dietary supplements and natural products as potential antiaging agents.


Intrinsically disordered domains: Sequence ➔ disorder ➔ function relationships.
Disordered domains are long regions of intrinsic disorder that ideally have conserved sequences, conserved disorder, and conserved functions. These domains were first noticed in protein-protein interactions that are distinct from the interactions between two structured domains and the interactions between structured domains and linear motifs or molecular recognition features (MoRFs). So far, disordered domains have not been systematically characterized. Here, we present a bioinformatics investigation of the sequence-disorder-function relationships for a set of probable disordered domains (PDDs) identified from the Pfam database. All the Pfam seed proteins from those domains with at least one PDD sequence were collected. Most often, if a set contains one PDD sequence, then all members of the set are PDDs or nearly so. However, many seed sets have sequence collections that exhibit diverse proportions of predicted disorder and structure, thus giving the completely unexpected result that conserved sequences can vary substantially in predicted disorder and structure. In addition to the induction of structure by binding to protein partners, disordered domains are also induced to form structure by disulfide bond formation, by ion binding, and by complex formation with RNA or DNA. The two new findings, (a) that conserved sequences can vary substantially in their predicted disorder content and (b) that homologues from a single domain can evolve from structure to disorder (or vice versa), enrich our understanding of the sequence ➔ disorder ensemble ➔ function paradigm.


The signalling conformation of the insulin receptor ectodomain.
Understanding the structural biology of the insulin receptor and how it signals is of key importance in the development of insulin analogs to treat diabetes. We report here a cryo-electron microscopy structure of a single insulin bound to a physiologically relevant, high-affinity version of the receptor ectodomain, the latter generated through attachment of C-terminal leucine zipper elements to overcome the conformational flexibility associated with ectodomain truncation. The resolution of the cryo-electron microscopy maps is 3.2 Å in the insulin-binding region and 4.2 Å in the membrane-proximal region. The structure reveals how the membrane proximal domains of the receptor come together to effect signalling and how insulin's negative cooperativity of binding likely arises. Our structure further provides insight into the high affinity of certain super-mitogenic insulins. Together, these findings provide a new platform for insulin analog investigation and design.


Circulating lysophosphatidylcholines are markers of a metabolically benign nonalcoholic fatty liver.
Nonalcoholic fatty liver (NAFL) is thought to contribute to insulin resistance and its metabolic complications. However, some individuals with NAFL remain insulin sensitive. Mechanisms involved in the susceptibility to develop insulin resistance in humans with NAFL are largely unknown. We investigated circulating markers and mechanisms of a metabolically benign and malignant NAFL by applying a metabolomic approach. A total of 265 metabolites were analyzed before and after a 9-month lifestyle intervention in plasma from 20 insulin-sensitive and 20 insulin-resistant subjects with NAFL. The relevant plasma metabolites were then tested for relationships with insulin sensitivity in 17 subjects without NAFL and in plasma from 29 subjects with liver tissue samples. The best separation of the insulin-sensitive from the insulin-resistant NAFL group was achieved by a metabolite pattern including the branched-chain amino acids leucine and isoleucine, ornithine, the acylcarnitines C3:0-, C16:0-, and C18:0-carnitine, and lysophosphatidylcholine (lyso-PC) C16:0 (area under the ROC curve, 0.77 [P = 0.00023] at baseline and 0.80 [P = 0.000019] at follow-up). Among the individual metabolites, predominantly higher levels of lyso-PC C16:0, both at baseline (P = 0.0039) and at follow-up (P = 0.001), were found in the insulin-sensitive compared with the insulin-resistant subjects. In the non-NAFL groups, no differences in lyso-PC C16:0 levels were found between the insulin-sensitive and insulin-resistant subjects, and these relationships were replicated in plasma from subjects with liver tissue samples. From a plasma metabolomic pattern, particularly lyso-PCs are able to separate metabolically benign from malignant NAFL in humans and may highlight important pathways in the pathogenesis of fatty liver-induced insulin resistance.


Bone marrow stroma in humans: anti-nerve growth factor receptor antibodies selectively stain reticular cells in vivo and in vitro.
Two anti-nerve growth factor receptor (LNGFR or p75NGFR) antibodies, Me20.4 and Me8211, label stromal cells with dendritic features in fresh smears and in formalin-fixed, paraffin-embedded human bone marrow (BM). The LNGFR+ cells have an oval nucleus, a scanty cytoplasm with long dendrites that intermingle with the hematopoietic cells, line the abluminal side of sinus endothelial cells, and provide the scaffold for the hematopoietic marrow. At the electron microscopy level, the immunogold tag labels the body and the long branching dendrites of fibroblast-like cells with scanty cytoplasm containing mitochondria, endoplasmic reticulum, and dense bodies. The LNGFR+ cells are positive for alkaline phosphatase, reticulin, collagen III, vimentin, TE-7, and CD13 but negative for endothelial (vWF, CD34, Pal-E), neural (CD56, neurofilament) and leukocyte markers (CD45, CD68). The LNGFR+ stromal cells appear in the fetal BM before the hematopoietic activity begins, originate from the vessel adventitia, and radiate in the Bm cavity. Long-term BM culture (LTBMC) in vitro contain LNGFR+ stromal cells. We document the presence of RNA message for the low- (LNGFR) and the high-affinity NGF receptor (NTRK1) by using RT-PCR on fresh BM aspirate and on LTBMC. BM biopsies from patients with hematologic fibrogenic diseases and in cytokine-treated cancer patients are evaluated for LNGFR+ cells: the amount of stained cells is correlated with the traditional reticulin stain in cases of myelofibrosis, therapy-related myelodysplasia, leukemia, and detected an increase of stromal cells in cytokine-treated patients. The anti-LNGFR antibodies represent a specific membrane marker for the adventitial reticular cells (ARC) of the human marrow and allow precise evaluation and quantitation of this important BM microenvironment component in vivo and in vitro.


Reduced Cost-sharing for Preventive Drugs Preferentially Benefits Low-income Patients With Diabetes in High Deductible Health Plans With Health Savings Accounts.
High deductible health plans linked to Health Savings Accounts (HSA-HDHPs) must include all care under the deductible except for select preventive services. Some employers and insurers have adopted Preventive Drug Lists (PDLs) that exempt specific classes of medications from deductibles. The objective of this study was to examine the association between shifts to PDL coverage and medication utilization among patients with diabetes in HSA-HDHPs. A natural experiment comparing pre-post changes in monthly and annual outcomes in matched study groups. The intervention group included 1744 commercially-insured HSA-HDHP patients with diabetes age 12-64 years switched by employers to PDL coverage; the control group included 3349 propensity-matched HSA-HDHP patients whose employers offered no PDL. Outcomes were out-of-pocket (OOP) costs for medications and the number of pharmacy fills converted to 30-day equivalents. Transition to the PDL was associated with a relative pre-post decrease of $612 (-35%, P<0.001) per member OOP medication expenditures; OOP reductions were higher for key classes of antidiabetic and cardiovascular medicines listed on the PDL; the policy did not affect unlisted classes. The PDL group experienced relative increases in medication use of 6.0 30-day fills per person during the year (+11.2%, P<0.001); the increase was more than twice as large for lower-income (+6.6 fills, +12.6%, P<0.001) than higher-income (+3.0 fills, +5.1%, P=0.024) patients. Transition to a PDL which covers important classes of medication to manage diabetes and cardiovascular conditions is associated with substantial annual OOP cost savings for patients with diabetes and increased utilization of important classes of medications, especially for lower-income patients.


"I don't have to know why it snows, I just have to shovel it!": Addiction Recovery, Genetic Frameworks, and Biological Citizenship.
The gene has infiltrated the way citizens perceive themselves and their health. However, there is scant research that explores the ways genetic conceptions infiltrate individuals' understanding of their own health as it relates to a behavioral trait, like addiction. Do people seeking treatment for addiction ground their self-perception in biology in a way that shapes their experiences? We interviewed 63 participants in addiction treatment programs, asking how they make meaning of a genetic understanding of addiction in the context of their recovery, and in dealing with the stigma of addiction. About two-thirds of people in our sample did not find a genetic conception of addiction personally useful to them in treatment, instead believing that the cause was irrelevant to their daily struggle to remain abstinent. One-third of respondents believed that an individualized confirmation of a genetic predisposition to addiction would facilitate their dealing with feelings of shame and accept treatment. The vast majority of our sample believed that a genetic understanding of addiction would reduce the stigma associated with addiction, which demonstrates the perceived power of genetic explanations in U.S. society. Our results indicate that respondents (unevenly) ground their self-perception of themselves as an addicted individual in biology.


Design criteria for transparent single-wall carbon nanotube thin-film transistors.
A study based on two-dimensional percolation theory yielding quantitative parameters for optimum connectivity of transparent single-wall carbon nanotube (SWNT) thin films is reported. Optimum SWNT concentration in the filtrated solution was found to be 0.1 mg/L with a volume of 30 mL. Such parameters lead to SWNT fractions in the films of approximately Phi = 1.8 x 10(-3), much below the metallic percolation threshold, which is found to be approximately PhiC = 5.5 x 10(-3). Therefore, the performance of transparent carbon nanotube thin-film transistors is limited by the metallic SWNTs, even below their percolation threshold. We show how this effect is related to hopping or tunneling between neighboring metallic tubes.


Fundoplication versus medical management of gastroesophageal reflux disease: systematic review and meta-analysis.
Medical, endoscopic, and open/laparoscopic surgical methods are used to treat gastroesophageal reflux disease (GERD). This study aimed to perform a systematic review of randomized controlled trials comparing medical and surgical treatments of GERD in adult patients. For the study, MEDLINE and EMBASE (1980-2012) were searched. Two reviewers independently assessed methodologic aspects and extracted data from eligible studies, focusing on patient-relevant outcomes. The primary outcomes were health-related and GERD-specific quality-of-life aspects. Standardized mean differences (SMDs) between treatment groups were calculated and combined using random-effect meta-analysis. The study identified 11 publications reporting on 7 trials comparing surgical (open or laparoscopic) and medical treatment of GERD. Meta-analysis of both quality-of-life aspects showed a pooled-effect estimate in favor of fundoplication (SMD 0.18; 95 % confidence interval [CI] 0.01-0.35; SMD 0.33; 95 % CI 0.13-0.54). Heartburn and regurgitation were less frequent after surgical intervention. However, a considerable proportion of patients still needed antireflux medication after fundoplication. Nevertheless, the surgical patients were significantly more satisfied with their symptom control and showed higher satisfaction with the treatment received. This systematic review showed that surgical management of GERD is more effective than medical management with respect to patient-relevant outcomes in the short and medium term. However, long-term studies still are needed to determine whether antireflux surgery is an equivalent alternative to lifelong medical treatment.


Evolution of bract development and B-class MADS box gene expression in petaloid bracts of Cornus s. l. (Cornaceae).
Despite increasing interest in the molecular mechanisms of floral diversity, few studies have investigated the developmental and genetic bases of petaloid bracts. This study examined morphological patterns of bract initiation and expression patterns of B-class MADS-box genes in bracts of several Cornus species. We suggest that petaloid bracts in this genus may not share a single evolutionary origin. Developmental pathways of bracts and spatiotemporal expression of B-class genes in bracts and flowers were examined for four closely related dogwood species. Divergent morphological progressions and gene expression patterns were found in the two sister lineages with petaloid bracts, represented by Cornus florida and Cornus canadensis. Phylogeny-based analysis identified developmental and gene expression changes that are correlated with the evolution of petaloid bracts in C. florida and C. canadensis. Our data support the existence of independent evolutionary origins of petaloid bracts in C. canadensis and C. florida. Additionally, we suggest that functional transference within B-class gene families may have contributed to the origin of bract petaloidy in C. florida. However, the underlying mechanisms of petaloid bract development likely differ between C. florida and C. canadensis. In the future this hypothesis can be tested by functional analyses of Cornus B-class genes.


High-speed multispectral videography with a periscope array in a spectral shaper.
We present a simple method for continuous snapshot multispectral imaging or multispectral videography that achieves high-speed spectral video recording without the need for mechanical scanning and much computation for datacube construction. The enabling component of this method is an array of periscopes placed in a prism-based spectral shaper that spectrally separates the image without image deformation. As a proof-of-principle demonstration, we show five-color multispectral video recording in the visible range (200×200 pixels per spectral image frame) at a record high frame rate of at least 2800 frames per second. Our experimental results indicate that this method holds promise for various industrial and biomedical applications such as remote sensing, food inspection, and endoscopy.


An evidence-based health workforce model for primary and community care.
The delivery of best practice care can markedly improve clinical outcomes in patients with chronic disease. While the provision of a skilled, multidisciplinary team is pivotal to the delivery of best practice care, the occupational or skill mix required to deliver this care is unclear; it is also uncertain whether such a team would have the capacity to adequately address the complex needs of the clinic population. This is the role of needs-based health workforce planning. The objective of this article is to describe the development of an evidence-informed, needs-based health workforce model to support the delivery of best-practice interdisciplinary chronic disease management in the primary and community care setting using diabetes as a case exemplar. Development of the workforce model was informed by a strategic review of the literature, critical appraisal of clinical practice guidelines, and a consensus elicitation technique using expert multidisciplinary clinical panels. Twenty-four distinct patient attributes that require unique clinical competencies for the management of diabetes in the primary care setting were identified. Patient attributes were grouped into four major themes and developed into a conceptual model: the Workforce Evidence-Based (WEB) planning model. The four levels of the WEB model are (1) promotion, prevention, and screening of the general or high-risk population; (2) type or stage of disease; (3) complications; and (4) threats to self-care capacity. Given the number of potential combinations of attributes, the model can account for literally millions of individual patient types, each with a distinct clinical team need, which can be used to estimate the total health workforce requirement. The WEB model was developed in a way that is not only reflective of the diversity in the community and clinic populations but also parsimonious and clear to present and operationalize. A key feature of the model is the classification of subpopulations, which gives attention to the particular care needs of disadvantaged groups by incorporating threats to self-care capacity. The model can be used for clinical, health services, and health workforce planning.


From Ideal to Real: Attachment Orientations Guide Preference for an Autonomous Leadership Style.
Autonomy is a key characteristic of attachment relations that varies as a function of attachment orientations and is also a key personality characteristic of leadership perceptions. In the presented research, we reasoned that the relationship between attachment and autonomy-related preference for specific leaders and leadership behavior would be a function of individuals' insecure attachment strategies. We tested our hypotheses in two studies. Study 1 used Multiple Indicators Multiple Causes (MIMIC) modeling to test expectations based on a cross-sectional design, while Study 2 utilized a vignette-based experimental design. We find that anxious individuals attributed less positive evaluations to an autonomous leadership style (Study 1), while avoidant persons attributed higher leader competence to an autonomous leader description (Study 2). Compared to less anxious participants, highly anxious participants attributed lower competence to the autonomous leader description. By examining how individual differences in attachment orientations can indirectly influence the ideal leader categorization process, the present set of studies lends support to the importance of attachment orientations and related working models in leader perception and contribute to the literature on leader-follower fit. Using a survey and experimental approach, we examine how followers' attachment schemas can shape the leader influence process, specifically concerning a preference for an autonomous leadership style.


Different Approaches to Meaning in Primate Gestural and Vocal Communication.
In searching for the roots of human language, comparative researchers investigate whether precursors to language are already present in our closest relatives, the non-human primates. As the majority of studies into primates' communication use a unimodal approach with focus on one signal type only, researchers investigate very different aspects depending on whether they are interested in vocal, gestural, or facial communication. Here, we focus on two signal types and discuss how meaning is created in the gestural (visual, tactile/auditory) as compared to the vocal modality in non-human primates, to highlight the different research foci across these modalities. First, we briefly describe the defining features of meaning in human language and introduce some debates concerning meaning in non-human communication. Second, with focus on these features, we summarize the current evidence for meaningful communication in gestural as compared to vocal communication and demonstrate that meaning is operationalized very differently by researchers in these two fields. As a result, it is currently not possible to generalize findings across these modalities. Rather than arguing for or against the occurrence of semantic communication in non-human primates, we aim at pointing to gaps of knowledge in studying meaning in our closest relatives, and these gaps might be closed.


Chronic isolation of carotid sinus baroreceptor region in conscious normotensive and hypertensive rats.
We have developed a chronic technique to isolate the carotid sinus baroreceptor region in the conscious rat model. Our technique, when used in conjunction with other methods, allows for the study of the control of arterial pressure, heart rate, and cardiac output by the carotid sinus baroreceptor reflex in conscious, unrestrained rats. The performance of our technique was evaluated in two strains: normotensive Sprague-Dawley (SD) rats and spontaneously hypertensive rats (SHR). Each rat was instrumented with an aortic flow probe and a catheter placed in the right femoral artery to monitor cardiac output and arterial pressure, respectively. The cervical sympathetic trunk and aortic depressor nerve were ligated and cut bilaterally, leaving vagus nerves intact. The right and left carotid sinuses were isolated using our new technique. We tested the open-loop function of the carotid sinus baroreceptor reflex system in the conscious rat after recovery from the isolation surgery. We found that changes in nonpulsatile carotid sinus pressure caused significant changes in arterial pressure, heart rate, and total peripheral resistance in both rat strains. However, the cardiac output responses differed dramatically between strains. Significant changes were seen in the cardiac output response of SHR, whereas no significant changes were observed in normotensive SD rats. We have found this technique to be a highly reliable tool for the study of the carotid sinus baroreceptor reflex system in the conscious rat.


Effect of displacement and distortion of potential energy surfaces and overlapping resonances of electronic transitions on surface-enhanced Raman scattering: models and ab initio theoretical calculation.
A previously developed theory for the temperature-dependent resonance Raman scattering is used to study the surface-enhanced Raman scattering. Two models, the displaced oscillator model and the displaced-distorted oscillator model, based on the harmonic potential energy surfaces are carried out to calculate the surface-enhanced Raman scattering excitation profiles of the pyridine molecule adsorbed on a silver electrode, for which the density functional theory method is applied to evaluate the potential energy surfaces of the adsorption structure. In this framework, the distortion effect on the surface-enhanced Raman scattering will be discussed by comparing both models. The overlapping resonance of multiexcited electronic transitions is also studied, in which the interference between electronic transitions has been taken into account. It will be used to study the abnormal band at 1005.6 cm(-1) with the exciting radiation 457.9 nm.


Coffee intake is associated with lower risk of symptomatic gallstone disease in women.
Metabolic studies have shown that coffee affects several hepatobiliary processes that are involved in cholesterol lithogenesis. We previously showed that coffee drinking was associated with a lower risk of symptomatic gallstone disease in men. We prospectively examined the association between coffee drinking and cholecystectomy, a surrogate of symptomatic gallstone disease, in a cohort of 80,898 women age 34-59 years in 1980 who had no history of gallstone disease. Coffee consumption and cholecystectomy were reported by participants on biennial mailed questionnaires. During 20 years of follow-up to the year 2000, 7,811 women reported a cholecystectomy. Compared with women who consistently reported consuming no caffeinated coffee, the multivariate relative risks (adjusting for risk factors for gallstone disease) of cholecystectomy comparing increasing categories of consistent intake of caffeinated coffee (0, 1, 2-3, and > or =4 cups/day) were 1.0, 0.91, 0.78, and 0.72 (95% confidence interval comparing extreme categories, 0.62-0.84; P value of test for trend < 0.0001). Caffeine intake from beverages and dietary sources was also inversely associated with risk of cholecystectomy. The multivariate relative risks comparing increasing categories of caffeine intake (< or =25, 26-100, 101-200, 201-400, 401-800, and >800 mg/day) were 1.0, 1.03, 1.01, 0.94, 0.85, and 0.85 (95% confidence interval comparing extreme categories, 0.74-0.96; P value of test for trend < 0.0001). In contrast, decaffeinated coffee was not associated with risk. These data suggest that consumption of caffeinated coffee may play a role in the prevention of symptomatic gallstone disease in women.


Infections of the fascial spaces of the neck.
Infections of fascial spaces of the neck continue to be life-threatening events. The frequency of these infections has been drastically reduced by antibiotics and good dental care. Intravenous drug use and immunosuppressed states have altered the modern presentation of these infections. This paper represents a review of these conditions, the anatomical and microbiological factors involved and the treatment. Attention is given to the use of modern imaging and contemporary antibiotic management. A clear understanding of regional anatomy remains critical to the proper resolution of these conditions.


Application of DNA barcoding to the identification of Hymenoptera parasitoids from the soybean aphid (Aphis glycines) in China.
Aphis glycines Matsumura is an important pest of soybean in Asia and North America. Hymenoptera parasitoids play a key role in the control of the soybean aphid. The correct identification of parasitoids is a critical step that precedes the assessment of their potential biological control agents. Accurate identification of the majority of the species attacking the soybean aphid often requires elaborate specimen preparation and expert taxonomic knowledge. In this study, we facilitated the identification of soybean aphid parasitoids by applying a DNA barcoding approach following a preliminary morphological identification. We generated DNA sequence data from the mitochondrial COI gene and the D2 region of 28S rDNA to assess the genetic variation within and between parasitoid species emerging from the soybean aphid in China. Fifteen Hymenoptera parasitoid species belonging to 10 genera of five families were identified with little intra-specific variation (0.09% ± 0.06% for 28S and 0.36% ± 0.18% for COI) and large inter-specific divergence (30.46% ± 3.42% for 28S and 20.4% ± 1.20% for COI).


Cardiopathy and ocular abnormalities in Noonan syndrome.
We describe three children with Noonan syndrome with cardiopathy. One female child had cardiopathy and ocular abnormalities. The other two male children had congenital heart disease of which one had uncommon association of tricuspid valve dysplasia with regurgitation associated with endocardial cushion defect. Karyotypes of the female and one of the male children were normal. The growth hormone and thyroid hormone studies in the first and second male children were normal. All the three children were managed conservatively and followed-up.


Quantification of cholinergic and select non-cholinergic mesopontine neuronal populations in the human brain.
The pars compacta and pars dissipata of the pedunculopontine nucleus contain cholinergic cell group Ch5, and the laterodorsal tegmental nucleus contains cholinergic cell group Ch6. The pedunculopontine nucleus has been implicated in a variety of functions, including mediation of rapid eye movement sleep and in extrapyramidal motor function, although the role of cholinergic and non-cholinergic neurons is unclear. Quantitative neuroanatomical techniques were used to map the distribution of cholinergic neurons in the mesopontine nuclei of the adult human brain. In addition, the number and distribution of comparably sized non-cholinergic neurons at selected anatomical levels were compared. An antibody raised against human choline acetyltransferase was used to stain immunohistochemically the mesopontine neurons in six brains, ranging in age from 28 to 60 years. The rostrocaudal length of the Cb5/Ch6 cell complex was approximately 10 mm. The estimated total number of cells was similar for all brains, and varied by less than 7%. The estimated average number of cholinergic cells in the combined pedunculopontine and laterodorsal tegmental nuclei was approximately 20,000, with 30% of the cells in the pedunculopontine nucleus pars compacta, 57% in the pedunculopontine nucleus pars dissipata and 13% in the laterodorsal tegmental nucleus. There was no correlation between cell number and age. Within areas of mesopontine tegmentum occupied by the Ch5 cholinergic neurons, there were often more noncholinergic neurons than comparably sized cholinergic neurons. The present study provides detailed maps of the distribution and number of mesopontine cholinergic neurons in the normal human brain. Many non-cholinergic neurons are intermixed with the cholinergic pedunculopontine neurons. One region of the pedunculopontine nucleus pars dissipata containing few cholinergic neurons, located adjacent to the ventral border of the pedunculopontine nucleus pars compacta, may correspond to the midbrain-extrapyramidal area as defined previously in rodent and in non-human primate. These data will be useful for quantitative neuropathological studies concerning the role of both cholinergic and non-cholinergic mesopontine neurons in diseases proposed to affect these neurons, including Parkinson's disease, schizophrenia and progressive supranuclear palsy.


Venous bypass in clinical liver transplantation.
A venous bypass technique (BP) that does not require the use of systemic anticoagulation is used routinely at our institution in all adult patients during the anhepatic phase of liver transplantation (LT). Complete cardiopulmonary profiles were obtained in a subset of 28 consecutive cases. During the anhepatic phase while on bypass, mean arterial pressure, central venous pressure, and pulmonary arterial wedge pressure were maintained at prehepatectomy levels. Oxygen consumption fell secondary to a decrease in temperature and the removal of the liver. Consequently, cardiac index fell without an increase in arterial-venous O2 content difference, reflecting adequate tissue oxygenation. Compared with 63 patients in a previous series given LT without bypass (NBP), the 57 total BP patients experienced better postoperative renal function (p less than 0.001), required less blood use during surgery (p less than 0.01), and had better survival 30 days after LT. The equivalency of 90-day survival in these groups results from the lack of effect of BP on the long-term survival of patients considered at high risk for metabolic reasons. BP patients at high risk for technical considerations, however, survived LT whereas NBP patients did not. BP offers other advantages important in establishing LT as a service-oriented procedure.


Serological diagnosis of paracoccidioidomycosis: high rate of inter-laboratorial variability among medical mycology reference centers.
Serological tests have long been established as rapid, simple and inexpensive tools for the diagnosis and follow-up of PCM. However, different protocols and antigen preparations are used and the few attempts to standardize the routine serological methods have not succeeded. We compared the performance of six Brazilian reference centers for serological diagnosis of PCM. Each center provided 30 sera of PCM patients, with positive high, intermediate and low titers, which were defined as the "reference" titers. Each center then applied its own antigen preparation and serological routine test, either semiquantitative double immunodifusion or counterimmmunoelectrophoresis, in the 150 sera from the other five centers blindly as regard to the "reference" titers. Titers were transformed into scores: 0 (negative), 1 (healing titers), 2 (active disease, low titers) and 3 (active disease, high titers) according to each center's criteria. Major discordances were considered between scores indicating active disease and scores indicating negative or healing titers; such discordance when associated with proper clinical and other laboratorial data, may correspond to different approaches to the patient's treatment. Surprisingly, all centers exhibited a high rate of "major" discordances with a mean of 31 (20%) discordant scores. Alternatively, when the scores given by one center to their own sera were compared with the scores given to their sera by the remaining five other centers, a high rate of major discordances was also found, with a mean number of 14.8 sera in 30 presenting a discordance with at least one other center. The data also suggest that centers that used CIE and pool of isolates for antigen preparation performed better. There are inconsistencies among the laboratories that are strong enough to result in conflicting information regarding the patients' treatment. Renewed efforts should be promoted to improve standardization of the serological diagnosis of PCM.


Evaluation of single whole inclusion serum test for IgG antibody to Chlamydia trachomatis in asymptomatic women.
IgG antibody titres against Chlamydia trachomatis were measured using a whole inclusion assay (Electro-Nucleonics Laboratories, Bethesda, Maryland, USA) in single serum samples of patients who presented for termination of pregnancy with evidence of infection with this organism (case group) and of 30 women with no evidence of infection (control group). Case and control patients were matched for age, race, marital status, history of sexually transmitted diseases, number of lifetime sexual partners, and whether they had had a new sexual partner in the preceding month. Fifty eight of 60 (97%) serum samples tested had chlamydial antibodies present at a titre of 1/8 or more. There was no significant difference between the geometric mean titres of the 30 case patients and the 28 controls with chlamydial antibodies (1/161.3 and 1/77.4 respectively). Significantly more case patients than controls, however, had a titre of 1/128 or more (p less than 0.01). The sensitivity, specificity, and positive and negative predictive values of a single serum antibody titre of 1/128 or more as an indicator of recent infection compared with isolation of the organism in cycloheximide treated McCoy cells or antigen detection by Chlamydiazyme (Abbot Laboratories; North Chicago, Illinois, USA), or both, were each 67%. The value of the antibody assay seems to be questionable as high levels of antibodies failed to correlate with isolation or antigen detection results, or both, in our population.


Flagella help Salmonella typhimurium survive within murine macrophages.
In this study, we evaluated how flagella enhance the pathogenicity of Salmonella typhimurium in strain C57BL/6J mice. When mice were infected orally with flagellated or nonflagellated S. typhimurium, equivalent numbers of bacteria colonized the gastrointestinal tracts of the animals, but the number of flagellated organisms increased faster once colonization began in the spleens and livers. To evaluate this differential rate of Salmonella growth, the rate of blood clearance, and the kinetics of net multiplication of salmonellae in splenic tissue after intravenous challenge, the two groups of mice were compared. We found that clearance of bacteria from the blood was the same for flagellated or nonflagellated strains. However, the number of flagellated bacteria in the spleen increased logarithmically until the death of the animals, whereas the number of nonflagellated salmonellae increased only slightly. In contrast, both flagellated and nonflagellated strains grew exponentially in the spleens of mice pretreated with silica, a macrophage toxic agent. In an in vitro macrophage assay, flagellated salmonellae survived longer than nonflagellated organisms. These results indicate that flagella either protect S. typhimurium from the intracellular killing mechanisms of murine macrophages or that flagella enhance the ability of S. typhimurium to multiply within murine macrophages.


What Counts is not Falling … but Landing: Strategic Analysis: An Adapted Model for Implementation Evaluation.
Implementation evaluations, also called process evaluations, involve studying the development of programmes, and identifying and understanding their strengths and weaknesses. Undertaking an implementation evaluation offers insights into evaluation objectives, but does not help the researcher develop a research strategy. During the implementation analysis of the UNAIDS drug access initiative in Chile, the strategic analysis model developed by Crozier and Friedberg was used. However, a major incompatibility was noted between the procedure put forward by Crozier and Friedberg and the specific characteristics of the programme being evaluated. In this article, an adapted strategic analysis model for programme evaluation is proposed.


Microbial thiosulphate reaction arrays: the interactive roles of Fe(III), O2 and microbial strain on disproportionation and oxidation pathways.
In this work, we experimentally evaluate pH and SO4(2-) dynamics associated with abiotic and microbial S2O3(2-) oxidation under varying [O2], [Fe(III)] and microbial strain/consortia (two pure strains, Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, their consortia, and two enrichments from an acidic environmental system, Moose Lake 2002 and Moose Lake 2003). Results of the batch experiments demonstrate highly active microbial processing of S2O3(2-) while abiotic controls under identical experimental conditions remain static with no pH decrease. When abiotic controls were manually titrated with acid to achieve similar pH decreases to those occurring in the microbial treatments, different S pathways were involved. In particular, disproportionation is a substantial component of initial microbial S2O3(2-) processing, and is accelerated by the presence of Fe(III), indicating that recycling of S through intermediate oxidation states is likely to be widespread in acidic mine environments where high [Fe(III)] is common. Furthermore, the microbially mediated S reaction pathways were dependent on both environmental conditions and microbial strain/consortia, indicating that microbial community structure also plays a key role. Collectively, these results highlight the importance of microbial activity, their poor representation by abiotic S models, the likelihood that Fe(III), rather than O2, is a key control on microbial S processing in acid environments and the need to identify the microbial community/strain involved.


Electromyographic evidence of selective fatigue during the eccentric phase of stretch/shortening cycles in man.
Ten male subjects were tested to determine the effects of muscle fatigue upon the activation pattern of the two main ankle extensor muscles, the 'slow-twitch' soleus (SOL) and the relatively 'fast-twitch' medial gastrocnemius (MG), during a fatiguing 60-s trial of hopping to maximal height. The myoelectric signals from SOL and MG were recorded together with the vertical ground reaction force signal and analysed by means of a computer-aided electromyograph (EMG) contour analysis, i.e. two-dimensional frequency distributions were obtained relating the activation patterns of the two synergists. The EMGs were also full-wave rectified and integrated (IEMG) according to three phases of the hopping movement (PRE, pre-activation phase; ECC, eccentric phase; CON, concentric phase). Results indicated that there were significant decreases (P less than 0.01) in the peak ground reaction force, the height of hopping and the mechanical power per unit body weight at the end of the fatiguing contractions. These decreases in mechanical parameters were accompanied by significant (P less than 0.01) decreases in all three phases of MG IEMG while SOL IEMG showed no such significant declines, except in the CON phase. Thus, the decreased mechanical parameters could in large part be accounted for by the substantial and selective decline of the excitation level of the relatively fast-twitch MG muscle. Our data suggest that the centrally mediated pre-activation of the fatiguable MG muscle as well as the MG activation during the eccentric phase, which is largely controlled by supraspinal inputs and stretch-reflex modulation, are most affected by fatigue changes during repeated maximal stretch/shortening cycles of the ankle extensors.


Unique genetic profiles from cerebrospinal fluid cell-free DNA in leptomeningeal metastases of EGFR-mutant non-small-cell lung cancer: a new medium of liquid biopsy.
Leptomeningeal metastases (LM) are more frequent in non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutations. Due to limited access to leptomeningeal lesions, the purpose of this study was to explore the potential role of cerebrospinal fluid (CSF) as a source of liquid biopsy in patients with LM. Primary tumor, CSF, and plasma in NSCLC with LM were tested by next-generation sequencing. In total, 45 patients with suspected LM underwent lumbar puncture, and those with EGFR mutations diagnosed with LM were enrolled. A total of 28 patients were enrolled in this cohort; CSF and plasma were available in 26 patients, respectively. Driver genes were detected in 100% (26/26), 84.6% (22/26), and 73.1% (19/26) of samples comprising CSF cell-free DNA (cfDNA), CSF precipitates, and plasma, respectively; 92.3% (24/26) of patients had much higher allele fractions in CSF cfDNA than the other two media. Unique genetic profiles were captured in CSF cfDNA compared with those in plasma and primary tissue. Multiple copy number variations (CNVs) were mainly identified in CSF cfDNA, and MET copy number gain identified in 47.8% (11/23) of patients was the most frequent one, while other CNVs included ERBB2, KRAS, ALK, and MYC. Moreover, loss of heterozygosity (LOH) of TP53 was identified in 73.1% (19/26) CSF cfDNA, which was much higher than that in plasma (2/26, 7.7%; P < 0.001). There was a trend towards a higher frequency of concomitant resistance mutations in patients with TP53 LOH than those without (70.6% versus 33.3%; P = 0.162). EGFR T790M was identified in CSF cfDNA of 30.4% (7/23) of patients who experienced TKI progression. CSF cfDNA could reveal the unique genetic profiles of LM and should be considered as the most representative liquid biopsy medium for LM in EGFR-mutant NSCLC.


Effects of blood viscosity on plasma renin activity and renal hemodynamics.
The effects of alterations in apparent blood viscosity on renal hemodynamics and plasma renin activity (PRA) were studied in dogs anesthetized with sodium pentobarbital. Blood viscosity was altered isovolemically either by changes in hematocrit (Hct) or by an increase in plasma viscosity (dextran administration). Arterial blood pressure and renal blood flow (RBF) remained relatively constant when apparent blood viscosity was elevated by changes in Hct or plasma viscosity. Thus the hyperviscosity of blood was associated with a decrease of renal vascular hindrance, resulting in an essentially unchanged renal flow resistance. The decrease in renal vascular hindrance may result from renal vasodilation. In hyperviscosity induced with dextran, the increase in PRA correlates linearly with the decrease in renal vascular hindrance, with a coefficient of correlation of 0.968 (P less than 0.005). The increase in PRA that resulted when Hct was raised from 25 to 55% also can be correlated linearly with the decrease in renal vascular hindrance, with a coefficient of correlation of 0.953 (P less than 0.005). These results suggest that the decrease in renal vascular hindrance in response to a rise in apparent blood viscosity leads to an increase in PRA.


Quantification of the esthetics of dentists' before and after photographs.
The purpose of this study was to measure dentists' before and after restorative treatment photographs to identify four metrics to describe the esthetic improvement: (a) central incisor width-to-height ratio; (b) central-to-lateral incisor width ratio; (c) lateral incisor-to-canine width ratio; and (d) lateral incisor percentage offset. Internet searches for "before after" and "veneers" and "prosthodontic" were used to obtain 198 before and after photographs of patient treatments showing the restoration of the six maxillary anterior teeth with porcelain veneers, crowns or a combination. The four metrics were measured using Adobe Photoshop. Groups were compared with repeated measures ANOVA followed by a post hoc Tukey-Kramer test with the variables of "Before or After Treatment"; "Treatment Type" (veneers, crowns or a combination) and "General Dentist or Prosthodontist". Mean central incisor width-to-height ratio was 91.7% before treatment, 80.8% after; mean central-to-lateral incisor width ratios were 69.9% and 64.7%; mean lateral incisor-to-canine width ratios were 85.3% and 81.4% and lateral incisor percentage offsets were 9.6% and 8.7%. There were significant (p<0.05) differences for before and after treatment for all variables except central-to-lateral incisor width ratio. Differences between specialist and general dentist were not statistically significant. On average, esthetic prosthetic treatment resulted in reduced central incisor width-to-height ratio, increased proportional width of the mesially positioned tooth of adjacent anterior teeth and reduced lateral incisor offset. The mean values of treatments by GPs and prosthodontists were not statistically significantly different. Knowledge of dentists' optimal restorative treatments provides insight on the esthetic outcome of extensive prosthodontic therapy.


Flow and wake characteristics associated with large wood to inform river restoration.
Wood is an integral part of a river ecosystem and the number of restoration projects using log placements is increasing. Physical model tests were used to explore how the wood position and submergence level (discharge) affect wake structure, and hence the resulting habitat. We observed a von-Kármán vortex street (VS) for emergent logs placed at the channel center, while no VS formed for submerged logs, because the flow entering the wake from above the log (sweeping flow) inhibited VS formation. As a result, emergent logs placed at the channel center resulted in ten times higher turbulent kinetic energy compared to submerged logs. In addition, both spatial variation in time-mean velocity and turbulence level increased with increasing log length and decreasing submergence level. Submerged logs and logs placed at the channel side created a greater velocity deficit and a longer recirculation zone, both of which can increase the residence time in the wake and deposition of organic matter and nutrients. The results demonstrate that variation in log size and degree of submergence can be used as a tool to vary habitat suitability for different fish preferences. To maximize habitat diversity in rivers, we suggest a diverse large wood placement.


Expression and localization of vascular endothelial growth factor A (VEGFA) and its two receptors (VEGFR1/FLT1 and VEGFR2/FLK1/KDR) in the canine corpus luteum and utero-placental compartments during pregnancy and at normal and induced parturition.
VEGFA is one of the most potent known inducers of angiogenesis. However, the function of angiogenic factors in the canine corpus luteum (CL) of pregnancy and in the pregnant uterus and placenta has not yet been elucidated. Therefore, here we investigated the expression and localization of VEGFA and its receptors (VEGFR1/FLT1 and VEGFR2/FLK1/KDR) in the canine CL and utero-placental compartments (ut-pl) throughout pregnancy until prepartum luteolysis. Antigestagen-mediated effects on expression of VEGF system in ut-pl were elucidated in mid-pregnant dogs. While displaying high individual variation, the luteal VEGFA was elevated during pre-implantation and post-implantation, followed by a decrease during mid-gestation, which was more pronounced at the mRNA level, and showed constant expression afterwards. Within the uterus, it increased following implantation and during mid-gestation in ut-pl compartments, but was downregulated at prepartum luteolysis. Luteal VEGFR1 expression resembled that of VEGFA; VEGFR2 remained unaffected throughout pregnancy. In ut-pl compartments, both receptors increased gradually towards mid-gestation; a prepartum decrease was observed for VEGFR1. Antigestagen-treatment resulted in decreased expression of ut-pl VEGFR1. In the CL, VEGFA stained in luteal cells. Uterine signals of VEGFA and its two receptors were observed in epithelial and vascular compartments, and in myometrium. In placental labyrinth, additionally, trophoblast stained positively. Luteal VEGFR1 was localized to the luteal cells and tunica media of blood vessels, whereas VEGFR2 stained only in capillary endothelial cells. The upregulation of luteal and the ut-pl VEGF system during early gestational stages supports the increased vascularization rate during this time. The diminishing effects of the prepartum endocrine milieu on VEGFA function seem to be more pronounced in the ut-pl units.


Patient follow-up in primary care after behavioral health screening in an urban public hospital system: a prospective, observational study of 2686 patients.
Patients with psychiatric conditions are known to experience poor and often disparate health outcomes. To investigate one potential mechanism for this phenomenon, we examined whether patients who screen positive for psychiatric comorbidity are lost to follow-up from primary care at higher rates than screen-negative controls. Patients in a public hospital system were followed prospectively for an 18-month period after an initial routine behavioral health screening in neighborhood health centers. Screening data were linked to electronic medical record visit data, and loss to follow-up was ascertained using Cox proportional hazards modeling. A public hospital health program screened 2686 patients from March 1998 to December 2000, and their visits were counted prospectively for 18 months. Nearly one third (N = 772, 29%) screened positive for a psychiatric condition. The screen-positive group had lower rates of censoring and a shorter time-to-event than the controls, indicating a higher continuing visit rate in primary care. This relationship persisted after adjustment for demographic variables, insurance type, substance abuse, and violence exposure. Patients who screen positive for psychiatric comorbidity are not lost to follow-up at higher rates than screen-negative controls. This finding suggests that disparate outcomes for mentally ill patients in a public hospital system may not be based on reduced access to or lack of contact with primary care providers. Further study of systems or provider-related factors is needed to ascertain the pathways toward poor health for this population.


Why are hatching and emergence success low? Mercury and selenium concentrations in nesting leatherback sea turtles (Dermochelys coriacea) and their young in Florida.
Leatherback sea turtles (Dermochelys coriacea) have low hatching and emergence success compared to other sea turtle species. Postmortem examinations of hatchlings showed degeneration of heart and skeletal muscle that was similar to that found in other neonates with selenium deficient mothers. Selenium deficiency can result from elevated concentrations of bodily mercury. Ingested mercury is detoxified by the liver through mercury-selenium compound formation. In animals persistently exposed to mercury, the liver's ability to detoxify this element may decrease, especially if dietary selenium is insufficient. We measured mercury and selenium concentrations in nesting female leatherbacks and their hatchlings from Florida and compared the levels to hatching and emergence success. Both liver selenium and the liver selenium-to-mercury ratio positively correlated with leatherback hatching and emergence success. This study provides the first evidence for the roles of mercury and selenium in explaining low reproductive success in a globally imperiled species, the leatherback sea turtle.


Combined QM/MM study of the mechanism and kinetic isotope effect of the nucleophilic substitution reaction in haloalkane dehalogenase.
Combined QM/MM molecular dynamics simulations have been carried out for the dehalogenation reaction of the nucleophilic displacement of dichloroethane catalyzed by haloalkane dehalogenase. The computed chlorine kinetic isotope effects and free energies of activation in the wild-type and the Phe172Trp mutant enzyme are found to be consistent with experiment. In comparison with the uncatalyzed model reaction in water, the enzyme lowers the activation barrier by about 16 kcal/mol. The enormous enzymatic action was attributed to a combination of contributions from a change in the solvation effect and transition state stabilization. The unique features of tryptophan's ability to interact favorably with hydrophobic substrates and to form hydrogen bonds to the leaving group chloride ion at the transition state enable both factors to make significant contributions to the barrier lowering mechanism in the enzyme. This is in contrast to the reference reaction in water, in which hydrogen bonding interactions are weakened at the transition state because of dispersed charge distribution at the transition state relative to that in the reactant and product states.


Mutations in LOXHD1, a recessive-deafness locus, cause dominant late-onset Fuchs corneal dystrophy.
Fuchs corneal dystrophy (FCD) is a genetic disorder of the corneal endothelium and is the most common cause of corneal transplantation in the United States. Previously, we mapped a late-onset FCD locus, FCD2, on chromosome 18q. Here, we present next-generation sequencing of all coding exons in the FCD2 critical interval in a multigenerational pedigree in which FCD segregates as an autosomal-dominant trait. We identified a missense change in LOXHD1, a gene causing progressive hearing loss in humans, as the sole variant capable of explaining the phenotype in this pedigree. We observed LOXHD1 mRNA in cultured human corneal endothelial cells, whereas antibody staining of both human and mouse corneas showed staining in the corneal epithelium and endothelium. Corneal sections of the original proband were stained for LOXHD1 and demonstrated a distinct increase in antibody punctate staining in the endothelium and Descemet membrane; punctate staining was absent from both normal corneas and FCD corneas negative for causal LOXHD1 mutations. Subsequent interrogation of a cohort of >200 sporadic affected individuals identified another 15 heterozygous missense mutations that were absent from >800 control chromosomes. Furthermore, in silico analyses predicted that these mutations reside on the surface of the protein and are likely to affect the protein's interface and protein-protein interactions. Finally, expression of the familial LOXHD1 mutant allele as well as two sporadic mutations in cells revealed prominent cytoplasmic aggregates reminiscent of the corneal phenotype. All together, our data implicate rare alleles in LOXHD1 in the pathogenesis of FCD and highlight how different mutations in the same locus can potentially produce diverse phenotypes.


Prostate-specific antigen and transrectal ultrasound of the prostate in detection of prostate cancer.
Detection of prostate cancer can be enhanced over the level historically obtained by digital rectal examination through the combined use of serum prostate-specific antigen and digital rectal examination, followed by transrectal ultrasonic examination of the prostate and ultrasonically guided biopsy if either of the initial studies is abnormal. Use of age-specific reference ranges for prostate-specific antigen results in greater sensitivity of this serum marker in patients below the age of 60 y and greater specificity in older patients.


A high degree of homology exists between the protein encoded by SLG and the S receptor domain encoded by SRK in self-incompatible Brassica campestris L.
The sporophytic self-incompatibility system in Brassica campestris is controlled by multiple alleles of a single locus, S. We isolated and characterized cDNA clones that correspond to genes for S-glycoprotein (SLG) and S-receptor kinase (SRK) from an S9-homozygous strain. An SRK9 fragment was amplified by the polymerase chain reaction (PCR) by use of primers that corresponded to regions of the gene for the extracellular receptor domain (S-domain) and the kinase domain. The fragment was used as a probe to isolate an SRK9 cDNA clone from the cDNA library of stigmas of an S9-homozygote. An SLG9 cDNA clone was also isolated from the library by use of SLG8 cDNA as a probe. Restriction fragment length polymorphism (RFLP) linkage analysis of the F2 plants from an S8S9 hybrid demonstrated that the SRK and SLG genes were tightly linked to one another and were also linked to S-genotypes, as determined by pollination tests. The transcripts of SRK9 and SLG9 were detected in stigmas, but not in anthers or leaves. The cytoplasmic kinase domain encoded by SRK9 contained amino acid sequences that are conserved in serine/threonine kinases. The nucleotide sequence encoding the S-domain of SRK9 was 98.4% homologous to that of SLG9 at the nucleotide level. This observation suggests the existence of a mechanism for maintaining a high degree of similarity between SLG and the region that encodes the S-domain of SRK.