Simultaneous control of glycemic, blood pressure, and lipid significantly reduce the risk of renal progression in diabetes patients.
Hyperglycemic, hypertension, and lipid abnormalities are risk factors for diabetic kidney disease However, no study has discussed the association of the simultaneous control of glycemic, blood pressure, and lipids with renal function among diabetes patients. Thus, this study examined the interactive effects of the intensive control of all 3 conditions on the progression of renal function. The study population was derived from eight hospitals in Taiwan from October 2008 to April 2015. Demographic characteristics were collected using structured questionnaires. Clinical variables were obtained from medical chart review. The renal progression was defined as a decline in the eGFR by more than 25% according to the baseline eGFR. Total of 1602 diabetes patients were included in the study analysis, the mean age was 63.03±10.98years, 55.56% were men. Compared to the simultaneous control of glycemic, blood pressure and lipid group, the poor control of all three diseases had the highest risk of renal progression, with an adjusted OR of 2.21 (95% CI, 1.26-3.86). Even if the patients with an intensive control of lipid, the result showed that the poor control of both glycemic and hypertension was associated with the increased risk of renal progression than the reference group. This study demonstrated that the simultaneous poor control of glycemic, blood pressure, and lipid had the highest risk of renal progression. Thus, patients with type 2 diabetes should not only control glycemic but also manage their blood pressure and lipid.


Stability and Performance of CsPbI2Br Thin Films and Solar Cell Devices.
In this manuscript, the inorganic perovskite CsPbI2Br is investigated as a photovoltaic material that offers higher stability than the organic-inorganic hybrid perovskite materials. It is demonstrated that CsPbI2Br does not irreversibly degrade to its component salts as in the case of methylammonium lead iodide but instead is induced (by water vapor) to transform from its metastable brown cubic (1.92 eV band gap) phase to a yellow phase having a higher band gap (2.85 eV). This is easily reversed by heating to 350 °C in a dry environment. Similarly, exposure of unencapsulated photovoltaic devices to water vapor causes current (JSC) loss as the absorber transforms to its more transparent (yellow) form, but this is also reversible by moderate heating, with over 100% recovery of the original device performance. NMR and thermal analysis show that the high band gap yellow phase does not contain detectable levels of water, implying that water induces the transformation but is not incorporated as a major component. Performances of devices with best efficiencies of 9.08% (VOC = 1.05 V, JSC = 12.7 mA cm-2 and FF = 68.4%) using a device structure comprising glass/ITO/c-TiO2/CsPbI2Br/Spiro-OMeTAD/Au are presented, and further results demonstrating the dependence of the performance on the preparation temperature of the solution processed CsPbI2Br films are shown. We conclude that encapsulation of CsPbI2Br to exclude water vapor should be sufficient to stabilize the cubic brown phase, making the material of interest for use in practical PV devices.


Electric field effects on nuclear magnetic shielding of the 1:1 and 2:1 (homo and heterochiral) complexes of XOOX' (X, X' = H, CH3) with lithium cation and their chiral discrimination.
The set of 1:1 and 2:1 complexes of XOOX' (X, X' = H, CH(3)) with lithium cation has been studied to determine if they are suitable candidates for chiral discrimination in an isotropic medium via nuclear magnetic resonance spectroscopy. Conventional nuclear magnetic resonance is unable to distinguish between enantiomers in the absence of a chiral solvent. The criterion for experimental detection is valuated by the isotropic part of nuclear shielding polarisability tensors, related to a pseudoscalar of opposite sign for two enantiomers. The study includes calculations at coupled Hartree-Fock and density functional theory schemes for (17)O nucleus in each compound. Additional calculations for (1)H are also included for some compounds. A huge static homogeneous electric field, perpendicular to the magnetic field of the spectromer, as big as ≈1.7 × 10(8) V m(-1) should be applied to observe a shift of ≈1 ppm for (17)O magnetic shielding in the proposed set of complexes.


A new bronchoscopic method to measure airway size.
Bronchoscopic evaluation of stenosis is limited due to radial distortion of bronchoscopic images and the unknown distance between the endoscope and the stenotic area. The purpose of this study was the development and validation of a method for measuring cross-sectional areas in large airways. Distance measurements were performed using a laser probe inserted into the working channel of a bronchoscope. The laser probe was positioned to the locus of interest in the airway, a ring of light (helium/neon) projected on to the luminal wall and the images acquired using an electronic bronchoscope. The images taken were distortion-corrected by means of a computer program. The method was validated by simulating airways using tubes of known diameter. Additionally, distortion-corrected bronchoscopic images were compared with distortion-free videoscopic image analysis of tracheal slices taken from pigs. In the case of the plastic tubes, Pearson's correlation coefficient (r) as well as the intraclass correlation coefficient (ICC) were slightly higher (r=0.99, p<0.01, ICC=0.97) than the correlation of cross-sectional areas between bronchoscopic and videoscopic images of tracheal slices (r=0.88, p<0.01, ICC=0.87). This concept allows accurate and reproducible determination of cross-sectional areas in large airways.


Secondary Electrostatic Interaction Model Revised: Prediction Comes Mainly from Measuring Charge Accumulation in Hydrogen-Bonded Monomers.
The secondary electrostatic interaction (SEI) model is often used to predict and explain relative hydrogen bond strengths of self-assembled systems. The SEI model oversimplifies the hydrogen-bonding mechanisms by viewing them as interacting point charges, but nevertheless experimental binding strengths are often in line with the model's predictions. To understand how this rudimentary model can be predictive, we computationally studied two tautomeric quadruple hydrogen-bonded systems, DDAA-AADD and DADA-ADAD. Our results reveal that when the proton donors D (which are electron-donating) and the proton acceptors A (which are electron-withdrawing) are grouped together as in DDAA, there is a larger accumulation of charge around the frontier atoms than when the proton donor and acceptor groups are alternating as in DADA. This accumulation of charge makes the proton donors more positive and the proton acceptors more negative, which enhances both the electrostatic and covalent interactions in the DDAA dimer. The SEI model is thus predictive because it provides a measure for the charge accumulation in hydrogen-bonded monomers. Our findings can be understood from simple physical organic chemistry principles and provide supramolecular chemists with meaningful understanding for tuning hydrogen bond strengths and thus for controlling the properties of self-assembled systems.


Three faces of recombination activating gene 1 (RAG1) mutations.
Severe combined immune deficiency (SCID) is a group of genetic disorder associated with development of T- and/or B-lymphocytes. Recombination-activating genes (RAG1/2) play a critical role on VDJ recombination process that leads to the production of a broad T-cell receptor (TCR) and B-cell receptor (BCR) repertoire in the development of T and B cells. RAG1/2 genes mutations result in various forms of primary immunodeficiency, ranging from classic SCID to Omenn syndrome (OS) to atypical SCID with such as granuloma formation and autoimmunity. Herein, we reported 4 patients with RAG1 deficiency: classic SCID was seen in two patients who presented with recurrent pneumonia and chronic diarrhoea, and failure to thrive. OS was observed in one patient who presented with chronic diarrhoea, skin rash, recurrent lower respiratory infections, and atypical SCID was seen in one patient who presented with Pyoderma gangrenosum (PG) and had novel RAG1 mutation.


Large orb-webs adapted to maximise total biomass not rare, large prey.
Spider orb-webs are the ultimate anti-ballistic devices, capable of dissipating the relatively massive kinetic energy of flying prey. Increased web size and prey stopping capacity have co-evolved in a number orb-web taxa, but the selective forces driving web size and performance increases are under debate. The rare, large prey hypothesis maintains that the energetic benefits of rare, very large prey are so much greater than the gains from smaller, more common prey that smaller prey are irrelevant for reproduction. Here, we integrate biophysical and ecological data and models to test a major prediction of the rare, large prey hypothesis, that selection should favour webs with increased stopping capacity and that large prey should comprise a significant proportion of prey stopped by a web. We find that larger webs indeed have a greater capacity to stop large prey. However, based on prey ecology, we also find that these large prey make up a tiny fraction of the total biomass (=energy) potentially captured. We conclude that large webs are adapted to stop more total biomass, and that the capacity to stop rare, but very large, prey is an incidental consequence of the longer radial silks that scale with web size.


Advancing translational research with the Semantic Web.
A fundamental goal of the U.S. National Institute of Health (NIH) "Roadmap" is to strengthen Translational Research, defined as the movement of discoveries in basic research to application at the clinical level. A significant barrier to translational research is the lack of uniformly structured data across related biomedical domains. The Semantic Web is an extension of the current Web that enables navigation and meaningful use of digital resources by automatic processes. It is based on common formats that support aggregation and integration of data drawn from diverse sources. A variety of technologies have been built on this foundation that, together, support identifying, representing, and reasoning across a wide range of biomedical data. The Semantic Web Health Care and Life Sciences Interest Group (HCLSIG), set up within the framework of the World Wide Web Consortium, was launched to explore the application of these technologies in a variety of areas. Subgroups focus on making biomedical data available in RDF, working with biomedical ontologies, prototyping clinical decision support systems, working on drug safety and efficacy communication, and supporting disease researchers navigating and annotating the large amount of potentially relevant literature. We present a scenario that shows the value of the information environment the Semantic Web can support for aiding neuroscience researchers. We then report on several projects by members of the HCLSIG, in the process illustrating the range of Semantic Web technologies that have applications in areas of biomedicine. Semantic Web technologies present both promise and challenges. Current tools and standards are already adequate to implement components of the bench-to-bedside vision. On the other hand, these technologies are young. Gaps in standards and implementations still exist and adoption is limited by typical problems with early technology, such as the need for a critical mass of practitioners and installed base, and growing pains as the technology is scaled up. Still, the potential of interoperable knowledge sources for biomedicine, at the scale of the World Wide Web, merits continued work.


Scales and drivers of seasonal pCO2 dynamics and net ecosystem exchange along the coastal waters of southeastern Arabian Sea.
The impact of seasonal coastal upwelling on the dynamics of dissolved inorganic carbon (DIC) and sea-air fluxes of CO2 along the coastal waters of Kochi was investigated during 2015, as a part of Ecosystem Modelling Project. The surface water pCO2 varied from 396 to 630μatm during the study period. Significant inter-seasonal variations were found in the distribution of physico-chemical variables and surface pCO2. An increase of 102.1μatm of pCO2 was noticed over a two-decade period with a rate of 5.3μatmy-1. There was an agreement between the fluxes of CO2 and net ecosystem production (NEP) with respect to the trophic status while NEP was higher than CO2 fluxes by a factor of 3.9. The annual net ecosystem exchange (NEE) was estimated to be 15.02mmolCm-2d-1 indicating that the coastal waters of Kochi are highly heterotrophic in nature.


In Situ Transplantation of Alginate Bioencapsulated Adipose Tissues Derived Stem Cells (ADSCs) via Hepatic Injection in a Mouse Model.
Adipose tissue derived stem cells (ADSCs) transplantation has recently gained widespread enthusiasm, particularly in the perspective to use them as potential alternative cell sources for hepatocytes in cell based therapy, mainly because of their capability of hepatogenic differentiation in vitro and in vivo. But some challenges remain to be addressed, including whether ADSCs can be provided effectively to the target organ and whether subsequent proliferation of transplanted cells can be achieved. To date, intrasplenic injection is the conventional method to deliver ADSCs into the liver; however, a number of donor cells retained in the spleen has been reported. In this study, our objective is to evaluate a novel route to transplant ADSCs specifically to the liver. We aimed to test the feasibility of in situ transplantation of ADSCs by injecting bioencapsulated ADSCs into the liver in mouse model. The ADSCs isolated from human alpha 1 antitrypsin (M-hAAT) transgenic mice were used to allow delivered ADSCs be readily identified in the liver of recipient mice, and alginate was selected as a cell carrier. We first evaluated whether alginate microspheres are implantable into the liver tissue by injection and whether ADSCs could migrate from alginate microspheres (study one). Once proven, we then examined the in vivo fate of ADSCs loaded microspheres in the liver. Specifically, we evaluated whether transplanted, undifferentiated ASDCs could be induced by the local microenvironment toward hepatogenic differentiation and the distribution of surviving ADSCs in major tissue organs (study two). Our results indicated ADSCs loaded alginate microspheres were implantable into the liver. Both degraded and residual alginate microspheres were observed in the liver up to three weeks. The viable ADSCs were detectable surrounding degraded and residual alginate microspheres in the liver and other major organs such as bone marrow and the lungs. Importantly, transplanted ADSCs underwent hepatogenic differentiation to become cells expressing albumin in the liver. These findings improve our understanding of the interplay between ADSCs (donor cells), alginate (biomaterial), and local microenvironment in a hepatectomized mouse model, and might improve the strategy of in situ transplantation of ADSCs in treating liver diseases.


Urine albumin excretion and subclinical cardiovascular disease. The Multi-Ethnic Study of Atherosclerosis.
We examined the association between urine albumin excretion (UAE) and common and internal carotid artery intima-media thickness (IMT), end-diastolic left ventricular (LV) mass, and coronary artery calcification (CAC) scores using data from the Multi-Ethnic Study of Atherosclerosis (MESA), a population-based study of 6814 adults aged 45 to 85 years without clinical cardiovascular disease (CVD). The mean age of the MESA participants was 62.7 years, 47% were male, and 15% had diabetes mellitus (DM). Sex-specific spot urine albumin/creatinine ratios were used to define 4 UAE categories: normal, high normal, microalbuminuria, and macroalbuminuria. CAC scores were log-transformed after adding 1 to all scores. Mean values of subclinical CVD measures were computed by level of UAE after adjustment for blood pressure, DM, and other covariates. After adjustment for all covariates, geometric mean CAC scores were higher among participants with high normal UAE (8.8; P=0.07), microalbuminuria (9.9; P=0.002), and macroalbuminuria (13.1; P=0.02) compared with normal UAE (7.4), but only microalbuminuria reached statistical significance. Mean LV mass (g/m2.7) was significantly higher in participants with high normal UAE (37.0; P=0.001), microalbuminuria (38.3; P< or =0.0001), and macroalbuminuria (42.3; P< or =0.0001) compared with normal UAE (36.0) after adjustment for all covariates. No significant difference in mean carotid IMT was found after adjustment for all covariates. Similar results were noted in MESA participants with and without DM. In conclusion, higher UAE, including levels below microalbuminuria, may reflect the presence of subclinical CVD among adults without established CVD.


Three-dimensional mapping of temporo-limbic regions and the lateral ventricles in schizophrenia: gender effects.
Local alterations in morphological parameters are poorly characterized in several brain regions widely implicated in schizophrenia neuropathology. Surface-based anatomical modeling was applied to magnetic resonance data to obtain three-dimensional (3D) average anatomical maps and measures of location, shape, asymmetry, and volume for the lateral ventricles, hippocampus, amygdala, and superior temporal gyrus in schizophrenic (n = 25; 15 male) and normal subjects (n = 28; 15 male) matched for demographic variables. For all regions, intra-group variability was visualized and group differences assessed statistically to discriminate local alterations in anatomy across sex and diagnosis. Posterior hippocampal volumes, lengths, and widths were reduced in patients. The right amygdala showed volume increases in schizophrenia patients versus controls. Ventricular enlargements, pronounced in the left hemisphere, occurred in the superior and lateral dimensions in patients, and these effects interacted with gender. Superior horn anterior extremes, inferior horn volumes, and hippocampal asymmetries exhibited gender effects. Significant group differences were absent in superior temporal gyrus parameters. Finally, regional variability profiles differed across groups. Clear morphometric differences of the lateral ventricles, hippocampus, and amygdala indicate regional displacements and shape distortions in several functional systems in schizophrenia. Alterations in these structures as mapped in 3D may provide the foundation for establishing brain abnormalities not previously defined at such a local level.


Circulating 25-hydroxyvitamin D and risk of pancreatic cancer: Cohort Consortium Vitamin D Pooling Project of Rarer Cancers.
Results from epidemiologic studies examining pancreatic cancer risk and vitamin D intake or 25-hydroxyvitamin D (25(OH)D) concentrations (the best indicator of vitamin D derived from diet and sun) have been inconsistent. Therefore, the authors conducted a pooled nested case-control study of participants from 8 cohorts within the Cohort Consortium Vitamin D Pooling Project of Rarer Cancers (VDPP) (1974-2006) to evaluate whether prediagnostic circulating 25(OH)D concentrations were associated with the development of pancreatic cancer. In total, 952 incident pancreatic adenocarcinoma cases occurred among participants (median follow-up, 6.5 years). Controls (n = 1,333) were matched to each case by cohort, age, sex, race/ethnicity, date of blood draw, and follow-up time. Conditional logistic regression analysis was used to calculate smoking-, body mass index-, and diabetes-adjusted odds ratios and 95% confidence intervals for pancreatic cancer. Clinically relevant 25(OH)D cutpoints were compared with a referent category of 50-<75 nmol/L. No significant associations were observed for participants with lower 25(OH)D status. However, a high 25(OH)D concentration (> or =100 nmol/L) was associated with a statistically significant 2-fold increase in pancreatic cancer risk overall (odds ratio = 2.12, 95% confidence interval: 1.23, 3.64). Given this result, recommendations to increase vitamin D concentrations in healthy persons for the prevention of cancer should be carefully considered.


Double-barrelled resuscitation: A feasibility and simulation study of dual-intraosseous needles into a single humerus.
Resuscitation can be delayed, or impaired, by insufficient vascular access. This study examines whether dual-intraosseous needles, inserted into a single porcine humerus, can facilitate rapid and concomitant fluid and medication delivery. After inserting one- and then two-intraosseous needles into the same porcine humerus, we determined the rate of fluid administration using (i) an infusion pump set to 999mL/h, and (ii) a standard pressure-bag set to 300mmHg. Next, we concomitantly infused blood, crystalloid and medications into the same medullary canal, using the two-needle set-up. Humeri were inspected for fluid-leakage, needle-displacement, and bone damage. Using an infusion pump, the mean normal-saline infusion-rate was significantly higher with dual-intraosseous needles compared to a single-intraosseous needle: the infusion-rate was 16mL/min using dual-needles versus 8mL/min for a single needle set-up (p<0.001). In contrast, using the pneumatic pressure-bag, the infusion rate was not statistically different when comparing dual-intraosseous needles versus single-intraosseous: the infusion-rate was 22mL/min versus 21ml/min (p=0.4) for 500mL, and 22ml/min versus 21ml/min (p=0.64) for one-litre, respectively. Blood product could be infused at a mean rate of 20mL/min through one needle while tranexamic acid was simultaneously infused through a second. There were no complications with a dual-intraosseous set-up (no fluid leakage; no needle-displacement; no high-pressure alarms, and no external bone-fractures or internal macrohistological damage) during any of our simulated resuscitation scenarios. This is the first published study evaluating dual-intraosseous needles in a single bone. Despite limitations, this preliminary study (using a porcine humerus) suggests that dual-intraosseous needles are feasible. For critically-ill patients with limited insertion sites, dual-intraosseous (a.k.a. 'double-barrelled resuscitation') may facilitate rapid and concurrent resuscitation.


Mechanisms underlying the anti-proliferative effects of berry components in in vitro models of colon cancer.
Consumption of fruit and vegetables is associated with a decreased risk of several cancers, particularly colorectal cancer, possibly linked to their phytochemical content, which is of interest due to several proposed health benefits, including potential anticancer activity. Epidemiological data suggests that cancers of the digestive tract are most susceptible to dietary modification, possibly due to being in direct contact with bioactive food constituents and therefore investigating the effects of these bioactive compounds on the prevalent colorectal cancer is feasible. Berries are a common element of Western diets, with members of the Rubus, Fragria, Sorbus, Ribes and Vaccinum genus featuring in desserts, preserves, yoghurts and juices. These soft fruit are rich in bioactive phytochemicals including several classes of phenolic compounds such as flavonoids (anthocyanins, flavonols and flavanols) and phenolic acids (hydroxybenzoic and hydroxycinnamic acids). Whilst there is little data linking berry consumption to reduced risk of colorectal cancer, in vitro evidence from models representing colorectal cancer suggests that berry polyphenols may modulate cellular processes essential for cancer cell survival, such as proliferation and apoptosis. The exact mechanisms and berry constituents responsible for these potential anticancer activities remain unknown, but use of in vitro models provides a means to elucidate these matters.


Analysis of BRCA1 variants in double-strand break repair by homologous recombination and single-strand annealing.
Missense substitutions of uncertain clinical significance in the BRCA1 gene are a vexing problem in genetic counseling for women who have a family history of breast cancer. In this study, we evaluated the functions of 29 missense substitutions of BRCA1 in two DNA repair pathways. Repair of double-strand breaks by homology-directed recombination (HDR) had been previously analyzed for 16 of these BRCA1 variants, and 13 more variants were analyzed in this study. All 29 variants were also analyzed for function in double-strand break repair by the single-strand annealing (SSA) pathway. We found that among the pathogenic mutations in BRCA1, all were defective for DNA repair by either pathway. The HDR assay was accurate because all pathogenic mutants were defective for HDR, and all nonpathogenic variants were fully functional for HDR. Repair by SSA accurately identified pathogenic mutants, but several nonpathogenic variants were scored as defective or partially defective. These results indicated that specific amino acid residues of the BRCA1 protein have different effects in the two related DNA repair pathways, and these results validate the HDR assay as highly correlative with BRCA1-associated breast cancer.


Effectiveness of an Integrated Community- and Clinic-Based Intervention on HIV Testing, HIV Knowledge, and Sexual Risk Behavior of Young Men Who Have Sex With Men in Myanmar.
Young men who have sex with men (YMSM) in Myanmar are disproportionately affected by HIV, with prevalence five times that of the general population. The Link Up project implemented an intervention using peer education and outreach providing education and counseling on health seeking around sexually transmitted infections and reproductive health, combined with focused clinic capacity building to improve the sexual and reproductive health of YMSM. This study aimed to evaluate the effectiveness and acceptability of the intervention. Using a mixed-methods approach, and employing a quasi-experimental design, we conducted two quantitative repeat cross-sectional surveys in purposively selected control (no intervention) and intervention townships, before and after implementation of the Link Up intervention. Respondent-driven sampling was used to recruit YMSM aged 15-24 years, and study participants were administered a structured questionnaire assessing intervention exposure, health service access, knowledge of HIV, and sexual risk behavior. Focus group discussions were held to elicit perspectives on the use and acceptability of the health services and peer outreach. At baseline, 314 YMSM were recruited in the intervention townships and 309 YMSM in the control townships. At end line, 267 (intervention) and 318 (control) YMSM were recruited. Coverage of the program was relatively low, with one-third of participants in the intervention townships having heard of the Link Up program by the end line. Comparing changes between baseline and end line, a greater proportion of HIV-negative or unknown status YMSM accessed HIV testing in the past 3 months in intervention townships (from 45.0% to 57.1%) compared with those in control townships (remained at 29.0%); however, this difference in the effect over time was not statistically significant in multivariate modeling (adjusted odds ratio: 1.45; 95% confidence interval: .66-3.17). Qualitative findings showed that the intervention was acceptable to YMSM. Overall, the intervention was perceived as acceptable. Although not statistically significant, results showed some trends toward improvements among YMSM in accessing HIV testing services and HIV-related knowledge. The modest coverage and short time frame of the evaluation likely limits the ability for any significant behavioral improvements.


Oral Biodisposition Study of Ritodrine after Its Buccal Administration in Rats.
In the previous study, buccal absorption of ritodrine (RD) hydrochloride was reported in vivo. As a result, buccal dosing of RD solution was found to be useful for the maintenance of effective plasma concentration. However, in order to find out the dosing schedule more clearly, it is important to clarify the in vivo drug behavior. The biodistributions of RD in oral cavity and buccal mucosal were investigated in order to understand the in vivo drug behavior after the buccal application. The pharmacokinetic parameters for 0 - infinite time and the absorption rate were calculated based on the plasma level-time profiles in the intravenous (1 mg/kg), buccal (10 mg/kg) and intragastric (10 mg/kg) dosings using rats. The drug concentrations in buccal mucosa and the remaining drug amounts in the oral cavity were examined over time after the buccal administration. From those drug distributions and drug absorption rates, the kinetic aspects were discussed. The absolute bioavailabilities of RD were 14.2% and 4.3% in buccal and intragastric (0.043) administrations, respectively. The oral cavity concentration was quickly eliminated within 0.5 h, and then decreased slowly. In both the administration site and distant region, the mucosal RD concentrations were observed at several dozen to approximately 100 μg/g during 0.5-4 h, indicating the rapid diffusion in the oral cavity. For both the mucosal parts, the buccal mucosal level reached a maximal level at 1 h, and then was slowly eliminated. The absorption rates were not related linearly to the buccal mucosal level, suggesting that the other mucosal parts such as sublingual mucosa and tongue ventral surface should be involved considerably in the absorption. The changes in RD concentration in oral cavity and oral mucosa showed the drug behavior in vivo. The present study revealed that RD is not accumulated in the buccal mucosa and transfers relatively fast from the oral mucosa to systemic circulation. It was suggested that the buccal dosing of RD should be acceptable even in the repeated manner as an alternative to the intravenous or oral administration of RD.


Phenotypic divergence of the common toad (Bufo bufo) along an altitudinal gradient: evidence for local adaptation.
Variation in the environment can induce different patterns of genetic and phenotypic differentiation among populations. Both neutral processes and selection can influence phenotypic differentiation. Altitudinal phenotypic variation is of particular interest in disentangling the interplay between neutral processes and selection in the dynamics of local adaptation processes but remains little explored. We conducted a common garden experiment to study the phenotypic divergence in larval life-history traits among nine populations of the common toad (Bufo bufo) along an altitudinal gradient in France. We further used correlation among population pairwise estimates of quantitative trait (QST) and neutral genetic divergence (FST from neutral microsatellite markers), as well as altitudinal difference, to estimate the relative role of divergent selection and neutral genetic processes in phenotypic divergence. We provided evidence for a neutral genetic differentiation resulting from both isolation by distance and difference in altitude. We found evidence for phenotypic divergence along the altitudinal gradient (faster development, lower growth rate and smaller metamorphic size). The correlation between pairwise QSTs-FSTs and altitude differences suggested that this phenotypic differentiation was most likely driven by altitude-mediated selection rather than by neutral genetic processes. Moreover, we found different divergence patterns for larval traits, suggesting that different selective agents may act on these traits and/or selection on one trait may constrain the evolution on another through genetic correlation. Our study highlighted the need to design more integrative studies on the common toad to unravel the underlying processes of phenotypic divergence and its selective agents in the context of environmental clines.


Robo4 is a vascular-specific receptor that inhibits endothelial migration.
Guidance and patterning of axons are orchestrated by cell-surface receptors and ligands that provide directional cues. Interactions between the Robo receptor and Slit ligand families of proteins initiate signaling cascades that repel axonal outgrowth. Although the vascular and nervous systems grow as parallel networks, the mechanisms by which the vascular endothelial cells are guided to their appropriate positions remain obscure. We have identified a putative Robo homologue, Robo4, based on its differential expression in mutant mice with defects in vascular sprouting. In contrast to known neuronal Robo family members, the arrangement of the extracellular domains of Robo4 diverges significantly from that of all other Robo family members. Moreover, Robo4 is specifically expressed in the vascular endothelium during murine embryonic development. We show that Robo4 binds Slit and inhibits cellular migration in a heterologous expression system, analogous to the role of known Robo receptors in the nervous system. Immunoprecipitation studies indicate that Robo4 binds to Mena, a known effector of Robo-Slit signaling. Finally, we show that Robo4 is the only Robo family member expressed in primary endothelial cells and that application of Slit inhibits their migration. These data demonstrate that Robo4 is a bona fide member of the Robo family and may provide a repulsive cue to migrating endothelial cells during vascular development.


Parkinson's disease-associated alterations of the gut microbiome predict disease-relevant changes in metabolic functions.
Parkinson's disease (PD) is a systemic disease clinically defined by the degeneration of dopaminergic neurons in the brain. While alterations in the gut microbiome composition have been reported in PD, their functional consequences remain unclear. Herein, we addressed this question by an analysis of stool samples from the Luxembourg Parkinson's Study (n = 147 typical PD cases, n = 162 controls). All individuals underwent detailed clinical assessment, including neurological examinations and neuropsychological tests followed by self-reporting questionnaires. Stool samples from these individuals were first analysed by 16S rRNA gene sequencing. Second, we predicted the potential secretion for 129 microbial metabolites through personalised metabolic modelling using the microbiome data and genome-scale metabolic reconstructions of human gut microbes. Our key results include the following. Eight genera and seven species changed significantly in their relative abundances between PD patients and healthy controls. PD-associated microbial patterns statistically depended on sex, age, BMI, and constipation. Particularly, the relative abundances of Bilophila and Paraprevotella were significantly associated with the Hoehn and Yahr staging after controlling for the disease duration. Furthermore, personalised metabolic modelling of the gut microbiomes revealed PD-associated metabolic patterns in the predicted secretion potential of nine microbial metabolites in PD, including increased methionine and cysteinylglycine. The predicted microbial pantothenic acid production potential was linked to the presence of specific non-motor symptoms. Our results suggest that PD-associated alterations of the gut microbiome can translate into substantial functional differences affecting host metabolism and disease phenotype.


mTOR independent alteration in ULK1 Ser758 phosphorylation following chronic LRRK2 kinase inhibition.
Unc-51 Like Kinase 1 (ULK1) is a critical regulator of the biogenesis of autophagosomes, the central component of the catabolic macroautophagy pathway. Regulation of ULK1 activity is dependent upon several phosphorylation events acting to repress or activate the enzymatic function of this protein. Phosphorylation of Ser758 ULK1 has been linked to repression of autophagosome biogenesis and was thought to be exclusively dependent upon mTOR complex 1 kinase activity. In the present study, a novel regulation of Ser758 ULK1 phosphorylation is reported following prolonged inhibition of the Parkinson's disease linked protein leucine rich repeat kinase 2 (LRRK2). Here, modulation of Ser758 ULK1 phosphorylation following LRRK2 inhibition is decoupled from the repression of autophagosome biogenesis and independent of mTOR complex 1 activity.


Nutrition management for critically and acutely unwell hospitalised patients with coronavirus disease 2019 (COVID-19) in Australia and New Zealand.
Coronavirus disease 2019 (COVID-19) results from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The clinical features and subsequent medical treatment, combined with the impact of a global pandemic, require specific nutritional therapy in hospitalised adults. This document aims to provide Australian and New Zealand clinicians with guidance on managing critically and acutely unwell adult patients hospitalised with COVID-19. These recommendations were developed using expert consensus, incorporating the documented clinical signs and metabolic processes associated with COVID-19, the literature from other respiratory illnesses, in particular acute respiratory distress syndrome, and published guidelines for medical management of COVID-19 and general nutrition and intensive care. Patients hospitalised with COVID-19 are likely to have preexisting comorbidities, and the ensuing inflammatory response may result in increased metabolic demands, protein catabolism, and poor glycaemic control. Common medical interventions, including deep sedation, early mechanical ventilation, fluid restriction, and management in the prone position, may exacerbate gastrointestinal dysfunction and affect nutritional intake. Nutrition care should be tailored to pandemic capacity, with early gastric feeding commenced using an algorithm to provide nutrition for the first 5-7 days in lower-nutritional-risk patients and individualised care for high-nutritional-risk patients where capacity allows. Indirect calorimetry should be avoided owing to potential aerosol exposure and therefore infection risk to healthcare providers. Use of a volume-controlled, higher-protein enteral formula and gastric residual volume monitoring should be initiated. Careful monitoring, particularly after intensive care unit stay, is required to ensure appropriate nutrition delivery to prevent muscle deconditioning and aid recovery. The infectious nature of SARS-CoV-2 and the expected high volume of patient admissions will require contingency planning to optimise staffing resources including upskilling, ensure adequate nutrition supplies, facilitate remote consultations, and optimise food service management. These guidelines provide recommendations on how to manage the aforementioned aspects when providing nutrition support to patients during the SARS-CoV-2 pandemic.


Participation in policy discourse: new form of exclusion for seniors with disabilities?
Recent discourses on aging emphasize the value of older people's social participation. How participation is defined in policy, however, may not correspond with seniors' realities. This article reports on the results of a critical discourse analysis conducted on aging policy in Quebec between 2005 and 2011. Results indicate that participation definitions can be problematic recommendations, standards, and expectations. Over time, participation increasingly came to be defined as productivity. The participation context also changed from collective responsibility to community adjustment and personal choice. Finally, policy texts reflected a polarization between activity and a loss of autonomy that linked participation with health status. Results suggest that, although innovative in the Canadian context, articulation of participation in Quebec's recent policies on aging lacks the politics from which to discuss difference, otherness, and access to participative opportunities. The case of older people's aging with disabilities illustrates the challenges of the new participatory agenda.


Repertoire of bovine miRNA and miRNA-like small regulatory RNAs expressed upon viral infection.
MicroRNA (miRNA) and other types of small regulatory RNAs play a crucial role in the regulation of gene expression in eukaryotes. Several distinct classes of small regulatory RNAs have been discovered in recent years. To extend the repertoire of small RNAs characterized in mammals and to examine relationship between host miRNA expression and viral infection we used Illumina's ultrahigh throughput sequencing approach. We sequenced three small RNA libraries prepared from cell line derived from the adult bovine kidney under normal conditions and upon infection of the cell line with Bovine herpesvirus 1. We used a bioinformatics approach to distinguish authentic mature miRNA sequences from other classes of small RNAs and short RNA fragments represented in the sequencing data. Using this approach we detected 219 out of 356 known bovine miRNAs and 115 respective miRNA* sequences. In addition we identified five new bovine orthologs of known mammalian miRNAs and discovered 268 new cow miRNAs many of which are not identifiable in other mammalian genomes and thus might be specific to the ruminant lineage. In addition we found seven new bovine mirtron candidates. We also discovered 10 small nucleolar RNA (snoRNA) loci that give rise to small RNA with possible miRNA-like function. Results presented in this study extend our knowledge of the biology and evolution of small regulatory RNAs in mammals and illuminate mechanisms of small RNA biogenesis and function. New miRNA sequences and the original sequencing data have been submitted to miRNA repository (miRBase) and NCBI GEO archive respectively. We envisage that these resources will facilitate functional annotation of the bovine genome and promote further functional and comparative genomics studies of small regulatory RNA in mammals.


Using null models to infer microbial co-occurrence networks.
Although microbial communities are ubiquitous in nature, relatively little is known about the structural and functional roles of their constituent organisms' underlying interactions. A common approach to study such questions begins with extracting a network of statistically significant pairwise co-occurrences from a matrix of observed operational taxonomic unit (OTU) abundances across sites. The structure of this network is assumed to encode information about ecological interactions and processes, resistance to perturbation, and the identity of keystone species. However, common methods for identifying these pairwise interactions can contaminate the network with spurious patterns that obscure true ecological signals. Here, we describe this problem in detail and develop a solution that incorporates null models to distinguish ecological signals from statistical noise. We apply these methods to the initial OTU abundance matrix and to the extracted network. We demonstrate this approach by applying it to a large soil microbiome data set and show that many previously reported patterns for these data are statistical artifacts. In contrast, we find the frequency of three-way interactions among microbial OTUs to be highly statistically significant. These results demonstrate the importance of using appropriate null models when studying observational microbiome data, and suggest that extracting and characterizing three-way interactions among OTUs is a promising direction for unraveling the structure and function of microbial ecosystems.


12.0-W continuous-wave diode-end-pumped Nd:GdVO(4) laser with high brightness operating at 912-nm.
We present a diode-pumped high-power continuous-wave (cw) laser at 912 nm based on the quasi-three-level (4)F(3/2)-->(4)4I(9/2) transition in Nd:GdVO(4) crystal. By using a 5-mm-long conventional bulk crystal with 0.2at.% Nd(3+)-doped concentration, the maximum output power of 12.0 W is obtained, with a slope efficiency of 29.3%. To the best of our knowledge, this is the highest output at 912 nm generated by diode-pumped Nd:GdVO(4) lasers. Furthermore, 3.2 W 456 nm deep-blue light is acquired by frequency doubling, resulting in an optical-to-optical efficiency of 8.0%. The short-term power instability of the blue laser is less than 3%.


Affinity binding of inclusion bodies on supermacroporous monolithic cryogels using labeling with specific antibodies.
A new chromatographic method based on affinity supermacroporous monolithic cryogels is developed for binding and analyzing inclusion bodies during fermentation. The work demonstrated that it is possible to bind specific IgG and IgY antibodies to the 15 and 17 amino acids at the terminus ends of a 33 kDa target protein aggregated as inclusion bodies. The antibody treated inclusion bodies from lysed fermentation broth can be specifically retained in protein A and pseudo-biospecific ligand sulfamethazine modified supermacroporous cryogels. The degree of binding of IgG and IgY treated inclusion bodies to the Protein A and sulfamethazine gels are investigated, as well as the influence of pH on the sulfamethazine ligand. Optimum binding of 78 and 72% was observed on both protein A and sulfamethazine modified cryogel columns, respectively, using IgG labeling of the inclusion bodies. The antibody treated inclusion bodies pass through unretained in the sulfamethazine supermacroporous gel at pH that does not favour the binding between the ligand on the gel and the antibodies on the surface of inclusion bodies. Also the unlabeled inclusion bodies went through the gel unretained, showing no non-specific binding or trapping within the gel. These findings may very well be the foundation for the building of a powerful analytical tool during fermentation of inclusion bodies as well as a convenient way to purify them from fermentation broth. These results also support our earlier findings [Kumar, A., Plieva, F.M., Galaev, I.Yu., Mattiasson, B., 2003. Affinity fractionation of lymphocytes using a monolithic cyogel. J. Immunol. Methods 283, 185-194] with mammalian cells that were surface labeled with specific antibodies and recognized on protein A supermacroporous gels. A general binding and separation system can be established on antibody binding cryogel affinity matrices.


Regulation of the DNA replication fork: a way to fight genomic instability.
DNA replication is a complex mechanism that functions due to the coordinated interplay of many factors. In the last few years, numerous studies have suggested that DNA replication factors are closely implicated in several DNA transaction events that maintain the integrity of the genome. Therefore, DNA replication fork factors have to be considered as part of a general process that aims to protect and replicate the genome in order to allow correct functioning of a cell and its eventual daughter cells. This is illustrated by the numerous factors that have a well-defined function at the DNA replication fork, but also play crucial roles in different DNA repair pathways such as base excision repair, nucleotide excision repair, double-strand break repair, and mismatch repair. Moreover, several of the replisome proteins have also been shown to be essential in sensing and transducing DNA damages through the checkpoint cascade pathways, including the recently characterised alternative clamps and clamp-loaders. In this review we present DNA replication factors that are involved in different DNA transaction and checkpoint regulation pathways, with emphasis on the link between DNA replication and maintenance of genomic stability.


Composite Polyelectrolyte Multilayer and Mesoporous Bioactive Glass Nanoparticle Coating on 316L Stainless Steel for Controlled Antibiotic Release and Biocompatibility.
Bacterial infection in wounds or implants can cause osteomyelitis, eventually leading to orthopedic implant failure. In this study, polyelectrolyte multilayer (PEM) coating comprising collagen as the cationic layer, chitosan as the barrier layer and γ-poly-glutamic acid as the anionic layer were fabricated onto a 316L stainless steel substrate by spin coating technique. Tetracycline-loaded 57S mesoporous bioactive glass nanoparticles (57S MBG, SiO₂:CaO:P₂O5 = 57:33:10 by wt%) were introduced into the γ-poly-glutamic acid layers. Herein, 57S MBG nanoparticles were successfully incorporated into the PEMs with a total thickness of ∼53 μm on 316L stainless steel (SS-PEMs-57S), which exhibited good hydrophilicity with a contact angle of 18.71°. The hardness of SS-PEMs-57S was 0.66 GPa while the Young's modulus was 11.5 GPa; these values are similar to those for the cortical bone. The surface roughness of MBG nanoparticle-incorporated PEMs increased from 231 to 384 nm. Controlled release of tetracycline loaded in MBG nanoparticles resulted in sustained antibacterial effect for up to 7 days, with higher release efficacy at low pH, which may be induced by inflammation or infection. Tetracycline loaded in SS-PEMs-57S showed good bacterial inhibition and maintained good cell viability in rat bone marrow mesenchymal stem cells (BMSCs) in the MTT assay. Moreover, SS-PEMs-57S also promoted mineralization of BMSCs. Therefore, this surface modification technology has great potential for endowing orthopedic implants with antibacterial and osteoconductive properties.


Is MPV a Predictive Marker for Neonatal Sepsis? A Pilot Study.
Neonatal sepsis (NS) continues to be a diagnostic challenge and a prime cause of mortality. Forage for a lucid, cost-effective yet highly sensitive and specific marker in diagnosing this entity is an incessant task. This study aimed to evaluate the predictive value of mean platelet volume (MPV) in diagnosing NS. Neonates diagnosed with sepsis from January 2016 to March 2016 were included in the study. The subjects were stratified into the following: (i) culture-proven sepsis (group I); (ii) culture-negative clinical sepsis (group II); and (iii) control group (group III). Several hematologic markers such as hemoglobin, total leukocyte count, platelet count, MPV, plateletcrit, platelet distribution width, immature-to-mature neutrophil ratio, toxic change, serum urea, bilirubin, and C-reactive protein were analyzed. The results were compared among the groups, and their efficacy in diagnosing NS was appraised. The study involved 210 neonates, of which, groups I, II, and III constituted 64, 75, and 71 cases, respectively. The mean MPV among groups I, II, and III was 9.56, 8.86, and 8.58 fL, respectively (P<0.05). Strikingly higher values of platelet count, immature-to-mature neutrophil ratio, MPV, plateletcrit, and C-reactive protein were found in group I in contrast to those in groups II and III (P<0.05). The baseline MPV of patients with culture-proven sepsis was comparatively higher than controls and was found to be statistically significant. Hence, MPV can be a simple, economical, and specific predictor of NS.


How much energy does the breast fed infant consume and expend?
Energy intake in breast fed infants is uncertain. The doubly labelled water method was used to measure, simultaneously and non-invasively, energy expenditure, energy intake, milk volume intake, energy deposition, and the energy content of breast milk in 12 "free living" breast fed babies at 5 and 11 weeks of age. The validity of this new approach was assessed in a parallel study in 12 formula fed infants. The babies who were exclusively breast fed expended 1.28 and 1.68 MJ/day at five and 11 weeks and had intakes of 1.81 and 2.22 MJ/day; these intakes were associated with normal growth but were well below those recommended previously. At five and 11 weeks the calculated energy content of breast milk was 0.24 and 0.25 MJ/100 ml, which is substantially lower than that commonly reported in milk obtained unphysiologically by expression of the breast. These data cast doubt on the widely used published standards for infant feeding.


Shining light on the antenna chromophore in lanthanide based dyes.
Lanthanide based dyes and assays exploit the antenna effect, where a sensitiser-chromophore is used as a light harvesting antenna and subsequent excited state energy transfer populates the emitting lanthanide centred excited state. A rudimentary understanding of the design criteria for designing efficient dyes and assays based on the antenna effect is in place. By preparing kinetically inert lanthanide complexes based on the DO3A scaffold, we are able to study the excited state energy transfer from a 7-methoxy-coumarin antenna chromophore to europium(iii) and terbium(iii) centred excited states. By contrasting the photophysical properties of complexes of metal centres with and without accessible excited states, we are able to separate the contributions from the heavy atom effect, photoinduced electron transfer quenching, excited state energy transfer and molecular conformations. Furthermore, by studying the photophysical properties of the antenna chromophore, we can directly monitor the solution structure and are able to conclude that excited state energy transfer from the chromophore singlet state to the lanthanide centre does occur.


Semicentralised supply and treatment systems: integrated infrastructure solutions for fast growing urban areas.
Currently, the development of the world population is characterised by two trends: absolute population growth and rapid urbanisation. Especially rapid urbanisation, taking place in Asia, Latin America and Africa, poses major pressure on the affected regions. The development of e.g. Asian countries today is stamped by a combination of urbanisation with high economic growth rates. Conventional centralised infrastructure of supply, treatment and disposal of water is not able to cope with the new challenges arising from these, in history incomparable, high growth rates. Therefore new approaches to infrastructure supply and treatment systems are required - for ecological, sociocultural and economic reasons. The semicentralised approach, focusing on integrated water supply and treatment structures for wastewater and waste on the neighbourhood level, offers one possible solution to the challenges imposed by rapid urbanisation and growing resource needs. The change from centralised to semicentralised supply and treatment systems will minimise the grave discrepancy between the rapid urban growth and the provision of supply and treatment infrastructure. Integrated semicentralised supply and treatment systems face the challenge of growing amounts of wastewater and solid waste combined with rising needs of water for private households and industrial use. The semicentralised approach offers a wide range of flexibility in implementation, energy self-sufficient operation, enormous saving potentials in water demands through intra-urban water reuse and further more advantages in comparison to centralised sectored solutions as practised today.


Pharmacologic treatment of dilated cardiomyopathy with special reference to the role of beta-blockers.
The pharmacologic treatment of dilated cardiomyopathy encompasses measures such as temporary administration of amino acids and calories for those who may be undernourished and avoidance of drugs such as alcohol, nicotine, amphetamines and high doses of tricyclic antidepressants. Anticoagulants should be used in patients with persistent failure or marked dilatation of the cardiac chambers since about 20% suffer from major embolic complications. In patients with ventricular tachycardia, R-on-T, or multifocal ventricular ectopic beats, class I antiarrhythmic drugs, or if unsuccessful, amiodarone should be administered since the risk of sudden death is substantial. In addition to digitalis and diuretics, the use of vasodilators is generally accepted to be of clinical value for reduction of symptoms in heart failure. Reports concerned with long-term clinical and hemodynamic effects with drugs such as prazosin, captopril and the combination of hydralazine and isosorbide dinitrate are conflicting and, so far, there is no evidence that any vasodilator prolongs survival. The beta-agonists, such as prenalterol, are limited in their usefulness by tachycardia induction, a tendency to provoke serious arrhythmias, tremor and anxiety as well as hemodynamic tolerance development. While the noncatecholamine inotropics such as milrinone may provide symptomatic relief in severe congestive heart failure, they also increase heart rate, induce ventricular arrhythmias and are associated with high one-year mortality rates. The beneficial effect of long-term beta-blockade, which we first reported in 1975 to include a decrease in functional class, increase in exercise tolerance, improvement in physical findings, increase in ejection fraction and enhanced survival, has been met with scepticism.(ABSTRACT TRUNCATED AT 250 WORDS)


Psychosocial interventions for women enrolled in alcohol treatment during pregnancy.
Excessive alcohol use during pregnancy has been associated with adverse maternal and neonatal effects. It is therefore important to develop and evaluate effective interventions during this important time in a woman's life. To our knowledge there have been no systematic reviews of randomised control trials (RCT) in this population. To evaluate the effectiveness of psychosocial interventions in pregnant women enrolled in alcohol treatment programs for improving birth and neonatal outcomes, maternal abstinence and treatment retention. We searched the Cochrane Drugs and Alcohol Group's Trial register (December 2007); MEDLINE (1950 to 2007); PsycINFO (1806 to 2007); EMBASE (1974 to 2007); CINAHL (1982 to 2007) We sought to include randomised or quasi-randomised studies comparing any psychosocial intervention versus pharmacological interventions or placebo or non-intervention or another psychosocial intervention for treating alcohol dependence in pregnancy. Three review authors independently assessed trials for inclusion in review. Studies were to be assessed using standardized data extraction and quality assessment forms. No suitable trials were identified. The search strategy identified 958 citations. 17 citations were deemed relevant for full text review, an additional 9 articles were retrieved through hand searching references, for a total of 26 articles. Following full text review no articles met the inclusion criteria. Data extraction and assessment of methodological quality were therefore not possible. The review question remains unanswered as there were no randomised control trials found relevant to the topic. There is a need for high quality randomised controlled trials to determine the effectiveness of psychosocial interventions in pregnant women enrolled in alcohol treatment programs.


All-in-one platform for salivary cotinine detection integrated with a microfluidic channel and an electrochemical biosensor.
Medical disorders caused by second-hand smoke are a major public health concern worldwide. To estimate the level of second-hand smoke exposure, salivary diagnostics for cotinine analysis is a compelling alternative in conventional diagnostics using bio-fluids, such as blood and urine, owing to its simple and non-invasive collection method. However, there are several critical issues, such as tedious multisteps, demand for expertise, and field unavailability to collect and transport the purified saliva for further analysis. Here, an all-in-one platform is presented to simply collect real human saliva and directly deliver it onto the biosensing surface. The platform consists of a commercial cotton-swab-type collector, 3D-printed housing, and microfluidic channel integrated with an electrochemical competitive immunosensor to evaluate the level of salivary cotinine. The immunosensor is based on a competitive binding assay between cotinine-conjugated horseradish peroxidase (C-HRP) and cotinine for anti-cotinine binding sites. The current responses obtained from the HRP-thionine-H2O2 system decreased proportionally to the cotinine concentration. This immunosensor successfully detected its target over a range of 1 × 10-1 to 1 × 104 pg ml-1 with a low limit of detection of 6 × 10-2 pg ml-1 and a limit of quantification of 1 × 10-1 pg ml-1. In addition, the platform is applicable to various commercial cotton-swab-type saliva collectors and can successfully transfer the saliva in wide flow rates ranging from 0.1 to 30 ml min-1 without leakage or damage to the sensing surface. Furthermore, the practicality of the proposed platform was evaluated by measuring cotinine in real human saliva from eight non-smokers. The concentration of cotinine was from 45.7 to 890.8 pg ml-1, which was in good agreement with that measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The introduced all-in-one platform represented a reliable performance delivering simple and practical steps in salivary diagnostics.


Validation of a portable pneumatic captive bolt device as a one-step method of euthanasia for use in depopulation of feedlot cattle.
To validate the effectiveness of a penetrating captive bolt device with a built-in low-pressure air channel pithing mechanism (PCBD) as a 1-step method for euthanasia of cattle. Clinical trial. 66 feedlot steers and heifers (weight, 227 to 500 kg [500 to 1,100 lb]) that were not expected to survive or finish the feeding period with their cohorts. Cattle were transported to a university facility and euthanized with the PCBD. For each calf, clinical variables were monitored and recorded immediately before and for at least 10 minutes after application of the PCBD. Following euthanasia, the head of each calf was removed and trauma to the brain and skull was assessed and scored. Death was successfully achieved with the PCBD without application of an ancillary technique in all 66 cattle; however, 4 (6%) cattle required a second or third shot from the PCBD because of technical errors in its placement. All shots from the PCBD that entered the cranial vault successfully rendered cattle unconscious without a return to sensibility. Results indicated that the PCBD was an effective 1-step method of euthanasia for use in mass depopulation of feedlot cattle.


Chemokine contribution to neuropathic pain: respective induction of CXCL1 and CXCR2 in spinal cord astrocytes and neurons.
Recent studies have indicated an important role of chemokines such as CCL2 in the development of chronic pain. However, the distinct roles of different chemokines in the development and maintenance of neuropathic pain and in their interactions with neurons have not been clearly elucidated. We found that spinal nerve ligation (SNL) not only induced persistent neuropathic pain symptoms, including mechanical allodynia and heat hyperalgesia, but also produced sustained CXCL1 upregulation in the spinal cord. Double staining of immunofluorescence and in situ hybridization revealed that CXCL1 was primarily induced in spinal astrocytes. In cultured astrocytes, tumor necrosis factor-α induced robust CXCL1 expression via the activation of the c-jun N-terminal kinase. Intrathecal administration of CXCL1 neutralizing antibody transiently reduced SNL-induced pain hypersensitivity, suggesting an essential role of CXCL1 in neuropathic pain sensitization. In particular, intraspinal delivery of CXCL1 shRNA lentiviral vectors, either before or after SNL, persistently attenuated SNL-induced pain hypersensitivity. Spinal application of CXCL1 not only elicited pain hypersensitivity but also induced rapid neuronal activation, as indicated by the expression of phosphorylated extracellular signal-regulated kinase and cAMP response element binding protein, and c-Fos in spinal cord neurons. Interestingly, CXCR2, the primary receptor of CXCL1, was upregulated in dorsal horn neurons after SNL, and the CXCR2 antagonist SB225002 completely blocked the CXCL1-induced heat hyperalgesia. SB225002 also attenuated SNL-induced pain hypersensitivity. Collectively, our results have demonstrated a novel form of chemokine-mediated glial-neuronal interaction in the spinal cord that can drive neuropathic pain. Inhibition of the CXCL1-CXCR2 signaling may offer a new therapy for neuropathic pain management.


Explicit dynamical electron-proton correlation in the nuclear-electronic orbital framework.
A method that includes explicit electron-proton correlation directly into the nuclear-electronic orbital self-consistent-field framework is presented. This nuclear-electronic orbital explicitly correlated Hartree-Fock (NEO-XCHF) scheme is formulated using Gaussian basis functions for the electrons and the quantum nuclei in conjunction with Gaussian-type geminal functions. The NEO approach is designed for the quantum treatment of a relatively small number of nuclei, such as the hydrogen nuclei involved in key hydrogen bonding interactions or hydrogen transfer reactions. The conventional nuclear-electronic-orbital-based methods produce nuclear wave functions that are too localized, leading to severe overestimations of hydrogen vibrational frequencies, as well as inaccuracies in geometries, isotope effects, couplings, and tunneling splittings. The application of the NEO-XCHF approach to a model system illustrates that the description of the nuclear wave function is significantly improved by the inclusion of explicit electron-proton correlation. In contrast to the NEO-HF method, the NEO-XCHF method leads to hydrogen vibrational stretch frequencies that are in excellent agreement with those calculated from grid-based methods. This approach is computationally practical for many-electron systems because only a relatively small number of nuclei are treated quantum mechanically and only electron-proton correlation is treated explicitly. Electron-electron dynamical correlation can be included with density functional theory or perturbation theory methods.


The autoimmune contrivance: genetics in the mouse model.
Autoimmunity and inheritance of complex characters behold an explosive interest in biology over the last 15 years. Research in the genetics of autoimmunity has been impelled by the isolation of genetic markers allowing tracing of heredity. The annotation and sequencing of the human and mouse genomes provide with the potential for further advancements, through the development of new technologies. This review aims to summarize advances made in the autoimmunity field, centered in type 1 diabetes in the NOD mouse model. It also aims to demonstrate that animal models, albeit some phenotypic and genetic dissimilarities with the human diseases, still remain the best way to move towards an understanding of the molecular mechanisms involved in autoimmunity. Assessing the current state of research in this field together with the increasing potential of novel biotechnology advancements, new insights to disease pathogenesis and discovery of molecular targets for intervention strategies are anticipated in the coming years.


Effect of prostaglandin E on immune function in multiple animal models.
The immunosuppression seen following major trauma and burns has long been attributed in part to prostaglandin E (PGE). This has been due primarily to the demonstration that PGE levels are elevated following burns and that when PGE is added to leukocyte cultures, it impairs multiple types of leukocyte functions. We investigated the effect of a new long-acting PGE derivative, 16,16-dimethyl-PGE, on immune function in multiple animal models. The PGE derivative had no effect on mortality in burn sepsis models but improved mean survival times in an Escherichia coli peritonitis model. The PGE derivative impaired neutrophil migration into burn wounds at lower dosages. In a rat burn model, when PGE was administered parenterally, it failed to impair cell-mediated immunity at any dosage and improved lymphocyte function at certain dosages. These data indicate that PGE may not be as immunosuppressive in in vivo models as it has been shown to be in in vitro models.


Artifacts in three-dimensional transesophageal echocardiography.
Three-dimensional (3D) transesophageal echocardiography (TEE) is subject to the same types of artifacts encountered on two-dimensional TEE. However, when displayed in a 3D format, some of the artifacts appear more "realistic," whereas others are unique to image acquisition and postprocessing. Three-dimensional TEE is increasingly used in the setting of percutaneous catheter-based interventions and ablation procedures, and 3D artifacts caused by the metallic components of catheters and devices are particularly frequent. Knowledge of these artifacts is of paramount relevance to avoid misinterpretation of 3D images. Although artifacts and pitfalls on two-dimensional echocardiography are well described and classified, a systematic description of artifacts in 3D transesophageal echocardiographic images and how they affect 3D imaging is still absent. The aim of this review is to describe the most relevant artifacts on 3D TEE, with particular emphasis on those occurring during percutaneous interventions for structural heart disease and ablation procedures.


An outbreak of Chandipura virus encephalitis in the eastern districts of Gujarat state, India.
An outbreak of encephalitis with a case fatality rate of 78.3% was investigated among children in Gujarat State, India. Twenty-six cases were reported. Three patients had IgM antibodies to Chandipura virus. Virus was isolated from one patient with rhabdomyosarcoma in porcine stable cell lines and in suckling mice. Chandipura virus RNA was present in 9 of 20 acute-phase serum samples, and virus sequences from the present outbreak were closely related to prototype strain (1965) and Andhra Pradesh, India (2003) isolates. Serologic and molecular assays documented the absence of Japanese encephalitis virus, West Nile virus, dengue virus, and paramyxoviruses in clinical samples. The etiologic agent was Chandipura virus, which has become an important encephalitis-causing virus in India.


Secular growth changes in the Hellenic population in the twentieth century.
Statural growth is dependent on hereditary and environmental factors, i.e disease, nutrition. The improvement of socioeconomic conditions that took place during the 20th century resulted in a secular trend towards greater height and earlier sexual maturation. Greek society has changed dramatically from a mainly agricultural society at the beginning of the 20th century to a mainly urban one in the second half of the century, and during this period Greece became a developed country. The various studies examining the height of children living in Athens during this period show a gradual increase in the height of children, the difference of the mean height between 2001 and 1928 being 11.8 cm and 7.3 cm for 17 year old boys and girls, respectively. The difference in mean height was present at all ages. The difference in final height was mainly due to prepubertal growth. Girls at the age of 10 and boys at 11 years were about 8 cm taller in 2001 than in 1928. A growth study carried out on conscripts in 1990 found no significant difference in the height of males coming from urban or rural areas of the country, whereas such a difference was detected in 1968, rural men being significantly shorter than urban ones in 1968. There are only a few studies on the sexual maturation of Greek children. The available data suggest a secular trend towards earlier puberty in females; however, this can not be substantiated for males. Menarcheal age in Greek girls showed a positive secular change that is in agreement with the observed trend for earlier pubertal maturation in girls. In conclusion, Greek children in the 20th century experienced a positive secular trend in stature which also includes final height. A secular trend for earlier sexual maturation can be shown only for girls.


Modulation of a pre-existing conformational equilibrium tunes adenylate kinase activity.
Structural plasticity is often required for distinct microscopic steps during enzymatic reaction cycles. Adenylate kinase from Escherichia coli (AK(eco)) populates two major conformations in solution; the open (inactive) and closed (active) state, and the overall turnover rate is inversely proportional to the lifetime of the active conformation. Therefore, structural plasticity is intimately coupled to enzymatic turnover in AK(eco). Here, we probe the open to closed conformational equilibrium in the absence of bound substrate with NMR spectroscopy and molecular dynamics simulations. The conformational equilibrium in absence of substrate and, in turn, the turnover number can be modulated with mutational- and osmolyte-driven perturbations. Removal of one hydrogen bond between the ATP and AMP binding subdomains results in a population shift toward the open conformation and a resulting increase of k(cat). Addition of the osmolyte TMAO to AK(eco) results in population shift toward the closed conformation and a significant reduction of k(cat). The Michaelis constants (K(M)) scale with the change in k(cat), which follows from the influence of the population of the closed conformation for substrate binding affinity. Hence, k(cat) and K(M) are mutually dependent, and in the case of AK(eco), any perturbation that modulates k(cat) is mirrored with a proportional response in K(M). Thus, our results demonstrate that the equilibrium constant of a pre-existing conformational equilibrium directly affects enzymatic catalysis. From an evolutionary perspective, our findings suggest that, for AK(eco), there exists ample flexibility to obtain a specificity constant (k(cat)/K(M)) that commensurate with the exerted cellular selective pressure.


Spermidine promotes stress resistance in Drosophila melanogaster through autophagy-dependent and -independent pathways.
The naturally occurring polyamine spermidine (Spd) has recently been shown to promote longevity across species in an autophagy-dependent manner. Here, we demonstrate that Spd improves both survival and locomotor activity of the fruit fly Drosophila melanogaster upon exposure to the superoxide generator and neurotoxic agent paraquat. Although survival to a high paraquat concentration (20 mM) was specifically increased in female flies only, locomotor activity and survival could be rescued in both male and female animals when exposed to lower paraquat levels (5 mM). These effects are dependent on the autophagic machinery, as Spd failed to confer resistance to paraquat-induced toxicity and locomotor impairment in flies deleted for the essential autophagic regulator ATG7 (autophagy-related gene 7). Spd treatment did also protect against mild doses of another oxidative stressor, hydrogen peroxide, but in this case in an autophagy-independent manner. Altogether, this study establishes that the protective effects of Spd can be exerted through different pathways that depending on the oxidative stress scenario do or do not involve autophagy.


Cost-effectiveness of a blended physiotherapy intervention compared to usual physiotherapy in patients with hip and/or knee osteoarthritis: a cluster randomized controlled trial.
Blended physiotherapy, in which physiotherapy sessions and an online application are integrated, might support patients in taking an active role in the management of their chronic condition and may reduce disease related costs. The aim of this study was to evaluate the cost-effectiveness of a blended physiotherapy intervention (e-Exercise) compared to usual physiotherapy in patients with osteoarthritis of hip and/or knee, from the societal as well as the healthcare perspective. This economic evaluation was conducted alongside a 12-month cluster randomized controlled trial, in which 108 patients received e-Exercise, consisting of physiotherapy sessions and a web-application, and 99 patients received usual physiotherapy. Clinical outcome measures were quality-adjusted life years (QALYs) according to the EuroQol (EQ-5D-3 L), physical functioning (HOOS/KOOS) and physical activity (Actigraph Accelerometer). Costs were measured using self-reported questionnaires. Missing data were multiply imputed and bootstrapping was used to estimate statistical uncertainty. Intervention costs and medication costs were significantly lower in e-Exercise compared to usual physiotherapy. Total societal costs and total healthcare costs did not significantly differ between groups. No significant differences in effectiveness were found between groups. For physical functioning and physical activity, the maximum probability of e-Exercise being cost-effective compared to usual physiotherapy was moderate (< 0.82) from both perspectives. For QALYs, the probability of e-Exercise being cost-effective compared to usual physiotherapy was 0.68/0.84 at a willingness to pay of 10,000 Euro and 0.70/0.80 at a willingness to pay of 80,000 Euro per gained QALY, from respectively the societal and the healthcare perspective. E-Exercise itself was significantly cheaper compared to usual physiotherapy in patients with hip and/or knee osteoarthritis, but not cost-effective from the societal- as well as healthcare perspective. The decision between both interventions can be based on the preferences of the patient and the physiotherapist. NTR4224 (25 October 2013).


Poly(ADP-ribose) polymerase regulates myocardial calcium handling in doxorubicin-induced heart failure.
Reactive oxygen and nitrogen species are overproduced in the cardiovascular system in response to the exposure to doxorubicin, a cardiotoxic anticancer compound. Oxidant-induced cell injury involves the activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) and pharmacological inhibition of PARP has recently been shown to improve myocardial contractility in doxorubicin-induced heart failure models. The current investigation, by utilizing an isolated perfused heart system capable of beat-to-beat intracellular calcium recording, addressed the following questions: (1) is intracellular calcium handling altered in hearts of rats after 6-week doxorubicin treatment, under baseline conditions, and in response to oxidative stress induced by hydrogen peroxide exposure in vitro; and (2) does pharmacological inhibition of PARP with the phenanthridinone-based PARP inhibitor PJ34 affect the changes in myocardial mechanical performance and calcium handling in doxorubicin-treated hearts under normal conditions and in response to oxidative stress. The results showed a marked elevation in intracellular calcium in the doxorubicin-treated hearts which was normalized by pharmacological inhibition of PARP. PARP inhibition also prevented the myocardial contractile disturbances and calcium overload that developed in response to hydrogen peroxide in the doxorubicin-treated hearts. We conclude that PARP activation contributes to the development of the disturbances in cellular calcium handling that develop in the myocardium in response to prolonged doxorubicin exposure.


Computational hemodynamic optimization predicts dominant aortic arch selection is driven by embryonic outflow tract orientation in the chick embryo.
In the early embryo, a series of symmetric, paired vessels, the aortic arches, surround the foregut and distribute cardiac output to the growing embryo and fetus. During embryonic development, the arch vessels undergo large-scale asymmetric morphogenesis to form species-specific adult great vessel patterns. These transformations occur within a dynamic biomechanical environment, which can play an important role in the development of normal arch configurations or the aberrant arch morphologies associated with congenital cardiac defects. Arrested migration and rotation of the embryonic outflow tract during late stages of cardiac looping has been shown to produce both outflow tract and several arch abnormalities. Here, we investigate how changes in flow distribution due to a perturbation in the angular orientation of the embryonic outflow tract impact the morphogenesis and growth of the aortic arches. Using a combination of in vivo arch morphometry with fluorescent dye injection and hemodynamics-driven bioengineering optimization-based vascular growth modeling, we demonstrate that outflow tract orientation significantly changes during development and that the associated changes in hemodynamic load can dramatically influence downstream aortic arch patterning. Optimization reveals that balancing energy expenditure with diffusive capacity leads to multiple arch vessel patterns as seen in the embryo, while minimizing energy alone led to the single arch configuration seen in the mature arch of aorta. Our model further shows the critical importance of the orientation of the outflow tract in dictating morphogenesis to the adult single arch and accurately predicts arch IV as the dominant mature arch of aorta. These results support the hypothesis that abnormal positioning of the outflow tract during early cardiac morphogenesis may lead to congenital defects of the great vessels due to altered hemodynamic loading.


Craniofacial trauma in injured motorcyclists: the impact of helmet usage.
Helmets are effective in decreasing maxillofacial trauma in motorcycle crashes. The impact, however, of motorcycle crashes on the location and patterns of craniofacial injuries among helmeted versus unhelmeted patients has not been examined. In the present study, 331 injured motorcyclists were evaluated to compare the incidence of craniofacial and spinal injury in 77 (23%) helmeted and 254 (77%) nonhelmeted patients. Nonhelmeted motorcyclists were three times more likely to suffer facial fractures (5.2% vs. 16.1%) than those wearing helmets (p < 0.01). Skull fracture occurred in only one helmeted patient (1.2%), compared with 36 (12.3%) of nonhelmeted patients (p < or = 0.01). The incidence of spinal injury was not significantly different between the two groups. Blood alcohol levels demonstrated that 12% of the helmeted group were legally intoxicated (blood alcohol level > 100 mg/dL), in contrast to 37.9% of the nonhelmeted motorcyclists (p < or = 0.01). Failure to wear a helmet resulted in a significantly higher incidence of craniofacial injury among patients involved in motorcycle crashes, but did not affect spinal injury or mortality. Alcohol usage seemed to correlate with failure to use helmets. Helmet use should be legally mandated on a national level for all motorcyclists.


[Development of a questionnaire to measure the effect of urinary incontinence on female sexuality: a reliability and validity study].
This study developed a measure of the degree of the effect of urinary incontinence on female sexuality and tested the reliability and validity of the developed measure. Effect of Urinary Incontinence on Sexuality Questionnaire (EISQ). The EISQ was developed from a thorough literature review and interviews of 20 individuals with urinary incontinence who said their incontinence had affected their dyadic relationship. These items were submitted to an analysis of content validity. This work resulted in a 28 items questionnaire that used a 4-point Likert-type response scale. The reliability and validity of the EISQ was tested with 144 women with incontinence who visited out-patient clinic in six hospitals. Cronbach's alpha as the index of internal consistency was high. The EISQ total was 0.90 and alpha ranged from 0.70 to 0.93 for the six subscales. Six factors each with an eigenvalue greater-than-or-equal-to-one were extracted from a principal components factors analysis with varimax rotation. The cumulative percentage of variance explained by this solution was 69.0%. Concurrent validity analyses confirmed our predictions that the EISQ scores were related to Patient Incontinence Stress (ISQ-P) scores. The EISQ proved to be reliable and valid as a self-administered measure for assessing the effect of urinary incontinence on female sexuality. The EISQ has demonstrated sufficient psychometric properties to warrant continued study of other samples and to explore the relationship between EISQ and other outcome variables.


Contextual effects in fine spatial discriminations.
The context in which a pattern is viewed can greatly affect its apparent contrast, a phenomenon commonly attributed to pooled contrast gain control processes. A low-contrast surround may slightly enhance apparent contrast, whereas increasing the contrast of the surround leads to a monotonic decline in contrast appearance. We ask here how the presence of a patterned surround affects the ability to perform fine, suprathreshold orientation, contrast, and spatial frequency discriminations as a function of surround contrast and phase. Our results revealed an unexpected dip in performance when center and surround were in phase and similar in contrast. These results suggest that additional processes, perhaps those involved in scene segregation, play a role in contextual effects on discrimination.


THE INCIDENCE OF BLACKHEAD AND OCCURRENCE OF HETERAKIS PAPILLOSA IN A FLOCK OF ARTIFICIALLY REARED TURKEYS.
In a flock of artificially reared turkeys originally consisting of 85 birds and reduced during the summer and fall by deaths and withdrawals for experimental purposes to 42 birds, five cases of blackhead occurred. These appeared during the months of July, September, and October. In four, Heterakis was searched for and found. In 38 birds from this flock killed for food during November and December, five harbored no Heterakis, and the rest carried light infestations. Of sixteen healthy birds withdrawn from the above flock during July and placed with a flock of older birds which had passed through this disease in former seasons, all contracted blackhead and fourteen died of the disease. The infestation with Heterakis was, as a rule, high, reaching a hundred specimens in some cases. In general, it appears that a high infestation with Heterakis is correlated with a high incidence of blackhead, a relation that had already been inferred in feeding experiments. In both of these groups no other species of worm was found in the ceca, and in instances in which examinations for coccidia were made none was found. Pheasants have been incriminated as a source of infestation with Heterakis papillosa in artificially reared flocks. In an artificially reared flock 38 birds that had never been ill, when killed in November and December, failed to show lesions of blackhead or evidence in the nature of scars that they had passed through an attack of the disease. Infectious soil that had remained unoccupied by turkeys and chickens for a period of 5 months beginning in the depth of a severe winter still harbored viable ova of Heterakis and proved highly dangerous to young poults. These experiments and observations fail to throw any light on the source of the protozoan parasite (Amoeba meleagridis) which causes the fatal lesions of blackhead.


Arthroscopic treatment of osteochondral lesions of the distal tibia.
Osteochondral lesions of the tibia are much less frequent than those of the talus, and treatment guidelines have not been established. We hypothesized that arthroscopic treatment methods used for osteochondral lesion of the talus would also be effective for those of the distal tibia. A review of 880 consecutive ankle arthroscopies identified 23 patients (2.6%) with osteochondral lesions of the distal tibia. Four patients were excluded because of concomitant acute ankle fractures requiring open reduction and internal fixation and two were lost to followup, leaving 17 in the study. The mean age was 38 (19 to 71) years. Six (35%) had osteochondral lesions of the tibia and talus; 11 had isolated lesions of the distal tibia. Treatment included excision, curettage, and abrasion arthroplasty in all patients. Five patients had transmalleolar drilling of the lesion, two had microfracture, and two had iliac bone grafting. At last followup, patients were evaluated with a questionnaire, physical examination, and ankle radiographs. Mean followup was 44 (24 to 99) months. Preoperatively, the median American Orthopaedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot score was 52; postoperatively, it was 87. Using the Wilcoxon signed-rank test to compare preoperative and postoperative scores, there was significant improvement in the ankle-hindfoot score postoperatively (p < 0.001). Seven patients had excellent results, seven had good results, one had a fair result, and two had poor results. Osteochondral lesions of the distal tibia present a challenge to the orthopedic surgeon. Arthroscopic treatment by means of debridement, curettage, abrasion arthroplasty, and, in some patients, transmalleolar drilling, microfracture, or iliac crest bone grafting, resulted in excellent and good results in 14 of 17 patients at medium-term followup.


Polarization-dependent extraction properties of bare fiber probes.
Despite their modest spatial resolution, uncoated tapered fiber probes are now widely used by the nano-optics community for mapping, with scanning near-field optical microscopy (SNOM), the nonradiative fields at the surface of optical and plasmonic microstructures and nanostructures. Given the significant complexity of the vectorial optical phenomena associated with subwavelength structures, the correct interpretation of SNOM acquisitions requires a complete and accurate understanding of the intrinsic image-formation procedure. In this theoretical study, we show that the SNOM imaging process with uncoated tapered fiber probes is highly polarization dependent and that the dominant effect is, surprisingly, the choice of optical fiber from which the tapered probe was fabricated. We demonstrate that although a tapered monomode fiber is unable to collect the component of the vector electric field parallel to the tip axis, a tapered multimode fiber can successfully collect all the three field components. However, we show that the signal from the longitudinal field component is collected only 10% as efficiently as the signal from the two transverse field components.


Impact of mitonuclear interactions on life-history responses to diet.
Mitochondria are central to both energy metabolism and biosynthesis. Mitochondrial function could therefore influence resource allocation. Critically, mitochondrial function depends on interactions between proteins encoded by the mitochondrial and nuclear genomes. Severe incompatibilities between these genomes can have pervasive effects on both fitness and longevity. How milder deficits in mitochondrial function affect life-history trade-offs is less well understood. Here, we analyse how mitonuclear interactions affect the trade-off between fecundity and longevity in Drosophila melanogaster. We consider a panel of 10 different mitochondrial DNA haplotypes against two contrasting nuclear backgrounds (w1118 (WE) and Zim53 (ZIM)) in response to high-protein versus standard diet. We report strikingly different responses between the two nuclear backgrounds. WE females have higher fecundity and decreased longevity on high protein. ZIM females have much greater fecundity and shorter lifespan than WE flies on standard diet. High protein doubled their fecundity with no effect on longevity. Mitochondrial haplotype reflected nuclear life-history trade-offs, with a negative correlation between longevity and fecundity in WE flies and no correlation in ZIM flies. Mitonuclear interactions had substantial effects but did not reflect genetic distance between mitochondrial haplotypes. We conclude that mitonuclear interactions can have significant impact on life-history trade-offs, but their effects are not predictable by relatedness. This article is part of the theme issue 'Linking the mitochondrial genotype to phenotype: a complex endeavour'.


The crystal structure of human deoxyhaemoglobin at 1.74 A resolution.
The structure of human deoxyhaemoglobin was refined at 1.74 A resolution using data collected on film at room temperature from a synchrotron X-ray source. The crystallographic R-factor is 16.0%. The estimated error in atomic positions is 0.1 A overall, 0.14 A for main-chain atoms of internal segments, and 0.05 A for the iron atoms. The effects of intermolecular contacts on the structure were investigated; such contacts cause only highly localized distortions, as judged from the degree of molecular asymmetry that they induce. The geometry of the iron-nitrogen complex closely resembles that of the deoxymyoglobin structure of Takano (1977) and of the 5-co-ordinated model compounds of Hoard (1975) and Jameson et al. (1980). The distance of the iron from the mean plane of N(porphyrin) is 0.40(5) A and 0.36(5) A, respectively, at the alpha and beta haems, in contrast to the corresponding distance of +0.12(8) A and -0.11(8) A in oxyhaemoglobin ( Shaanan , 1983); the Fe-N epsilon (F8) bond length is 2.12(4) A and the Fe-N(porphyrin) bond length is 2.06(2) A; the last is also in good agreement with extended X-ray fluorescence spectroscopy measurements on deoxyhaemoglobin ( Eisenberger et al., 1978; Perutz et al., 1982). The haems are domed toward the proximal side; the separation between the mean planes of N(porphyrin) and C(porphyrin) being 0.16(6) A and 0.10(6) A, respectively at the alpha and beta haems. At the alpha haems, the normals to the mean pyrrole planes are tilted uniformly toward the haem centre, by about three degrees relative to the haem normal, and there is a folding of about four degrees of the haem about an axis running between the methene carbons that are between the pyrrole rings bearing like-type side-chains. At the beta haems, there is no such folding, and only pyrroles II and IV (those eclipsed by His F8) are appreciably tilted, by about eight degrees. The independence of these parameters from restraints imposed on the model was verified by unrestrained refinement of the entire molecule starting from a structure with modified haem geometry.


Adiponectin-mediated changes in effector cells involved in the pathophysiology of rheumatoid arthritis.
Rheumatoid arthritis (RA) is associated with increased production of adipokines, which are cytokine-like mediators that are produced mainly in adipose tissue but also in synovial cells. Since RA synovial fibroblasts (RASFs), lymphocytes, endothelial cells, and chondrocytes are key players in the pathophysiology of RA, this study was undertaken to analyze the effects of the key adipokine adiponectin on proinflammatory and prodestructive synovial effector cells. Lymphocytes were activated in part prior to stimulation. All cells were stimulated with adiponectin, and changes in gene and protein expression were determined by Affymetrix and protein arrays. Messenger RNA and protein levels were confirmed using semiquantitative reverse transcription-polymerase chain reaction (PCR), real-time PCR, and immunoassays. Intracellular signal transduction was evaluated using chemical signaling inhibitors. Adiponectin stimulation of human RASFs predominantly induced the secretion of chemokines, as well as proinflammatory cytokines, prostaglandin synthases, growth factors, and factors of bone metabolism and matrix remodeling. Lymphocytes, endothelial cells, and chondrocytes responded to adiponectin stimulation with enhanced synthesis of cytokines and various chemokines. Additionally, chondrocytes released increased amounts of matrix metalloproteinases. In RASFs, adiponectin-mediated effects were p38 MAPK and protein kinase C dependent. Our previous findings indicated that adiponectin was present in inflamed synovium, at sites of cartilage invasion, in lymphocyte infiltrates, and in perivascular areas. The findings of the present study indicate that adiponectin induces gene expression and protein synthesis in human RASFs, lymphocytes, endothelial cells, and chondrocytes, supporting the concept of adiponectin being involved in the pathophysiologic modulation of RA effector cells. Adiponectin promotes inflammation through cytokine synthesis, attraction of inflammatory cells to the synovium, and recruitment of prodestructive cells via chemokines, thus promoting matrix destruction at sites of cartilage invasion.


An adipoinductive role of inflammation in adipose tissue engineering: key factors in the early development of engineered soft tissues.
Tissue engineering and cell implantation therapies are gaining popularity because of their potential to repair and regenerate tissues and organs. To investigate the role of inflammatory cytokines in new tissue development in engineered tissues, we have characterized the nature and timing of cell populations forming new adipose tissue in a mouse tissue engineering chamber (TEC) and characterized the gene and protein expression of cytokines in the newly developing tissues. EGFP-labeled bone marrow transplant mice and MacGreen mice were implanted with TEC for periods ranging from 0.5 days to 6 weeks. Tissues were collected at various time points and assessed for cytokine expression through ELISA and mRNA analysis or labeled for specific cell populations in the TEC. Macrophage-derived factors, such as monocyte chemotactic protein-1 (MCP-1), appear to induce adipogenesis by recruiting macrophages and bone marrow-derived precursor cells to the TEC at early time points, with a second wave of nonbone marrow-derived progenitors. Gene expression analysis suggests that TNFα, LCN-2, and Interleukin 1β are important in early stages of neo-adipogenesis. Increasing platelet-derived growth factor and vascular endothelial cell growth factor expression at early time points correlates with preadipocyte proliferation and induction of angiogenesis. This study provides new information about key elements that are involved in early development of new adipose tissue.


Criminal Justice Involvement and Service Need among Men on Methadone who Have Perpetrated Intimate Partner Violence.
Perpetrators of male-to-female intimate partner violence (IPV) may be likely to have multiple service needs, the extent of which may vary with respect to criminal justice involvement. The salience of the criminal justice system and the potential impact on service needs due to arrest and incarceration is underscored given the association between substance use and IPV. This study utilized a sample of men in methadone treatment who perpetrated male-to-female IPV in order to examine associations between criminal justice involvement and perceived additional service need(s). Results indicate that the likelihood of having a service need(s) significantly increased as time since most recent arrest or incarceration decreased. These findings highlight the need and potential benefit that can be derived from greater coordination amongst the criminal justice, IPV prevention, and drug treatment systems and service providers.