Microfluidics for cryopreservation.
Minimizing cell damage throughout the cryopreservation process is critical to enhance the overall outcome. Osmotic shock sustained during the loading and unloading of cryoprotectants (CPAs) is a major source of cell damage during the cryopreservation process. We introduce a microfluidic approach to minimize osmotic shock to cells during cryopreservation. This approach allows us to control the loading and unloading of CPAs in microfluidic channels using diffusion and laminar flow. We provide a theoretical explanation of how the microfluidic approach minimizes osmotic shock in comparison to conventional cryopreservation protocols via cell membrane transport modeling. Finally, we show that biological experiments are consistent with the proposed mathematical model. The results indicate that our novel microfluidic-based approach improves post-thaw cell survivability by up to 25% on average over conventional cryopreservation protocols. The method developed in this study provides a platform to cryopreserve cells with higher viability, functionality, and minimal inter-technician variability. This method introduces microfluidic technologies to the field of biopreservation, opening the door to future advancements at the interface of these fields.


Projector-Based Embedding Eliminates Density Functional Dependence for QM/MM Calculations of Reactions in Enzymes and Solution.
Combined quantum mechanics/molecular mechanics (QM/MM) methods are increasingly widely utilized in studies of reactions in enzymes and other large systems. Here, we apply a range of QM/MM methods to investigate the Claisen rearrangement of chorismate to prephenate, in solution, and in the enzyme chorismate mutase. Using projector-based embedding in a QM/MM framework, we apply treatments up to the CCSD(T) level. We test a range of density functional QM/MM methods and QM region sizes. The results show that the calculated reaction energetics are significantly more sensitive to the choice of density functional than they are to the size of the QM region in these systems. Projector-based embedding of a wave function method in DFT reduced the 13 kcal/mol spread in barrier heights calculated at the DFT/MM level to a spread of just 0.3 kcal/mol, essentially eliminating dependence on the functional. Projector-based embedding of correlated ab initio methods provides a practical method for achieving high accuracy for energy profiles derived from DFT and DFT/MM calculations for reactions in condensed phases.


Occupational exposures and lung cancer risk among Minnesota taconite mining workers.
To examine the association between employment duration, elongate mineral particle (EMP) exposure, silica exposure and the risk of lung cancer in the taconite mining industry. We conducted a nested case-control study of lung cancer within a cohort of Minnesota taconite iron mining workers employed by any of the mining companies in operation in 1983. Lung cancer cases were identified by vital records and cancer registry data through 2010. Two age-matched controls were selected from risk sets of cohort members alive and lung cancer free at the time of case diagnosis. Calendar time-specific exposure estimates were made for every job and were used to estimate workers' cumulative exposures. ORs and 95% CIs were estimated using conditional logistic regression. We evaluated total lung cancer risk and risk of histological subtype by total work duration and by cumulative EMP, and silica exposure by quartile of the exposure distribution. A total of 1706 cases and 3381 controls were included in the analysis. After adjusting for work in haematite mining, asbestos exposure and sex, the OR for total duration of employment was 0.99 (95% CI 0.96 to 1.01). The ORs for quartile 4 versus 1 of EMP and silica exposure were 0.82 (95% CI 0.57 to 1.19) and 0.97 (95% CI 0.70 to 1.35), respectively. The risk of each histological subtype of lung cancer did not change with increasing exposure. This study suggests that the estimated taconite mining exposures do not increase the risk of developing lung cancer.


Anti-biofilm properties of chitosan-coated surfaces.
Surfaces coated with the naturally-occurring polysaccharide chitosan (partially deacetylated poly N-acetyl glucosamine) resisted biofilm formation by bacteria and yeast. Reductions in biofilm viable cell numbers ranging from 95% to 99.9997% were demonstrated for Staphylococcus epidermidis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Candida albicans on chitosan-coated surfaces over a 54-h experiment in comparison to controls. For instance, chitosan-coated surfaces reduced S. epidermidis surface-associated growth more than 5.5 (10)log units (99.9997%) compared to a control surface. As a comparison, coatings containing a combination of the antibiotics minocycline and rifampin reduced S. epidermidis growth by 3.9 (10)log units (99.99%) and coatings containing the antiseptic chlorhexidine did not significantly reduce S. epidermidis surface associated growth as compared to controls. The chitosan effects were confirmed with microscopy. Using time-lapse fluorescence microscopy and fluorescent-dye-loaded S. epidermidis, the permeabilization of these cells was observed as they alighted on chitosan-coated surfaces. This suggests chitosan disrupts cell membranes as microbes settle on the surface. Chitosan offers a flexible, biocompatible platform for designing coatings to protect surfaces from infection.


A vanadium-doped ZnO nanosheets-polymer composite for flexible piezoelectric nanogenerators.
We report high performance flexible piezoelectric nanogenerators (PENGs) by employing vanadium (V)-doped ZnO nanosheets (NSs) and the polydimethylsiloxane (PDMS) composite structure. The V-doped ZnO NSs were synthesized to overcome the inherently low piezoelectric properties of intrinsic ZnO. Ferroelectric phase transition induced in the V-doped ZnO NSs contributed to significantly improve the performance of the PENGs after the poling process. Consequently, the PENGs exhibited high output voltage and current up to ∼32 V and ∼6.2 μA, respectively, under the applied strain, which are sufficient to directly turn on a number of light emitting diodes (LEDs). The composite approach for PENG fabrication is scalable, robust, and reproducible during periodic bending/releasing over extended cycles. The approach introduced here extends the performance limits of ZnO-based PENGs and demonstrates their potential as energy harvesting devices.


Acute care and long-term mortality among elderly patients with intracerebral hemorrhage who undergo chronic life-sustaining procedures.
Little is known about patients with intracerebral hemorrhage (ICH) who undergo chronic life-sustaining procedures. We sought to explore variations in treatment, Medicare payments, and mortality among elderly patients with ICH who received a feeding tube, a tracheostomy, or neither chronic life-sustaining procedure. Medicare Provider Analysis and Review files from 2004 linked to Center for Medicaid and Medicare Services denominator files through January 2005 were analyzed. Patients over age 65 years with a primary diagnosis of ICH based on discharge code (ICD-9-CM 431) were divided into those who underwent tracheostomy, those who underwent feeding tube placement but not tracheostomy, and those who underwent neither procedure. Thirty-day and 1-year survival rates were estimated using Kaplan-Meier methods. Among the 32,210 patients studied, 6% underwent feeding tube placement, and 2.5% underwent tracheostomy. Compared with the patients who did not undergo a chronic life-sustaining procedure, those who underwent tracheostomy had a longer length of stay (median, 25 days vs 4 days; P < .01) and greater Medicare spending (median, $81,479 vs $6,008; P < .01) during their initial hospitalization. The 30-day and 1-year cumulative mortality risks were 47% and 59%, respectively, in patients who did not undergo a chronic life-sustaining procedure, 21% and 53% in patients who underwent feeding tube placement, and 19% and 65% in those who underwent tracheostomy (P < .01, log-rank test across the 3 groups). Our findings show high 1-year mortality among elderly patients with ICH, even in those who undergo chronic life-sustaining procedures. Medicare payments for patients who undergo tracheostomy are substantial. More information about functional outcomes is needed.


Polishing ponds for post-treatment of digested sewage. Part 1: Flow-through ponds.
Polishing ponds are used to improve the quality of effluents from efficient anaerobic sewage treatment plants like UASB reactors, so that the final effluent quality becomes compatible with legal or desired standards. The residual organic material and suspended solids concentrations in the digested sewage are reduced, but often the main objective of polishing ponds is to improve the hygienic quality, measured by the concentration of two indicator organisms: helminth eggs and faecal coliforms (FC). The FC removal is normally the slowest process and for that reason becomes the main design criterion for a polishing pond. By contrast in conventional waste stabilisation pond (WSP) systems the organic material removal is the governing design parameter. The feasibility of operating a single polishing pond for the post-treatment of UASB effluent is shown in this paper and the final effluent quality as a function of the retention time is discussed. Even under the most adverse weather conditions (several weeks of rain) the population of algae remained stable and produced enough oxygen to maintain a predominantly aerobic environment. The final effluent TSS and BOD concentrations were not very low for retention times of less than 1 week, but this could be attributed to the presence of algae in the final effluent. Filtered effluent BOD and TSS concentrations were very low. For retention times of more than 1 week algae were efficiently removed from the liquid phase by the action of predators and algae flocculation and settling, so that a final effluent with a very low BOD and TSS concentrations was produced. To maximise the FC removal efficiency the polishing pond was constructed with the objective of approaching a plug flow regime. However, the observed efficiency was well below the expected value for all retention times, which was attributable to imperfections of the flow regime. From tracer studies it was established that the dispersion number was in the range of 0.14 to 0.16, which means that moderate mixing occurred, even though the pond was designed to avoid mixing as much as possible. Also the dead volume fraction ranging from 0.12 to 0.15 was quite considerable. Due to these imperfections the required retention time for an effluent to be used in unrestricted irrigation was produced for a retention time of about 10 days, twice the value of the minimum retention time for a batch or true plug flow pond. Although the plug flow regime could not be approached in practice, the required retention time of 10 days is still very much shorter than the value used in conventional WSPs (20 to 30 days).


Rise and fall of the Beringian steppe bison.
The widespread extinctions of large mammals at the end of the Pleistocene epoch have often been attributed to the depredations of humans; here we present genetic evidence that questions this assumption. We used ancient DNA and Bayesian techniques to reconstruct a detailed genetic history of bison throughout the late Pleistocene and Holocene epochs. Our analyses depict a large diverse population living throughout Beringia until around 37,000 years before the present, when the population's genetic diversity began to decline dramatically. The timing of this decline correlates with environmental changes associated with the onset of the last glacial cycle, whereas archaeological evidence does not support the presence of large populations of humans in Eastern Beringia until more than 15,000 years later.


Fullerene growth from encapsulated graphene flakes.
The direct in situ observation of fullerene formation encapsulated within a graphene ridge has been made possible using an aberration corrected transmission electron microscope (AC-TEM). An atom-by-atom mechanism was proposed based on in situ AC-TEM observations. First principle calculations found a continuous energy decrease upon the addition of carbon atoms to the edge of the graphene flakes, which mimics the fullerene growth steps and supports the atom-by-atom mechanism. The ridged graphene structure worked as a container for pinning small graphene flakes and capturing carbon atoms, which increased the growth probability of the fullerene structure within the small encapsulated space.


Horizontal Transposon Transfer and Its Implications for the Ancestral Ecology of Hydrophiine Snakes.
Transposable elements (TEs), also known as jumping genes, are sequences able to move or copy themselves within a genome. As TEs move throughout genomes they often act as a source of genetic novelty, hence understanding TE evolution within lineages may help in understanding environmental adaptation. Studies into the TE content of lineages of mammals such as bats have uncovered horizontal transposon transfer (HTT) into these lineages, with squamates often also containing the same TEs. Despite the repeated finding of HTT into squamates, little comparative research has examined the evolution of TEs within squamates. Here we examine a diverse family of Australo-Melanesian snakes (Hydrophiinae) to examine if the previously identified, order-wide pattern of variable TE content and activity holds true on a smaller scale. Hydrophiinae diverged from Asian elapids ~30 Mya and have since rapidly diversified into six amphibious, ~60 marine and ~100 terrestrial species that fill a broad range of ecological niches. We find TE diversity and expansion differs between hydrophiines and their Asian relatives and identify multiple HTTs into Hydrophiinae, including three likely transferred into the ancestral hydrophiine from fish. These HTT events provide the first tangible evidence that Hydrophiinae reached Australia from Asia via a marine route.


Gliotransmission of D-serine promotes thirst-directed behaviors in Drosophila.
Thirst emerges from a range of cellular changes that ultimately motivate an animal to consume water. Although thirst-responsive neuronal signals have been reported, the full complement of brain responses is unclear. Here, we identify molecular and cellular adaptations in the brain using single-cell sequencing of water-deprived Drosophila. Water deficiency primarily altered the glial transcriptome. Screening the regulated genes revealed astrocytic expression of the astray-encoded phosphoserine phosphatase to bi-directionally regulate water consumption. Astray synthesizes the gliotransmitter D-serine, and vesicular release from astrocytes is required for drinking. Moreover, dietary D-serine rescues aay-dependent drinking deficits while facilitating water consumption and expression of water-seeking memory. D-serine action requires binding to neuronal NMDA-type glutamate receptors. Fly astrocytes contribute processes to tripartite synapses, and the proportion of astrocytes that are themselves activated by glutamate increases with water deprivation. We propose that thirst elevates astrocytic D-serine release, which awakens quiescent glutamatergic circuits to enhance water procurement.


CST for the grand finale of telomere replication.
Telomeric DNA at eukaryotic chromosome ends terminates with single stranded 3' G-rich overhangs. The overhang is generated by the interplay of several dynamic processes including semiconservative DNA replication, 3' end elongation by telomerase, C-strand fill-in synthesis and nucleolytic processing. The mammalian CST (CTC1-STN1-TEN1) complex is directly involved at several stages of telomere end formation. Elucidation of its structural organization and identification of interaction partners support the notion that mammalian CST is, as its yeast counterpart, a RPA-like complex. CST binding at mammalian telomere 3' overhangs increases upon their elongation by telomerase. Formation of a trimeric CST complex at telomeric 3'overhangs leads to telomerase inhibition and at the same time mediates a physical interaction with DNA polymerase-α. Thus CST seems to play critical roles in coordinating telomerase elongation and fill-in synthesis to complete telomere replication.


Probing the Cellular Size Distribution in Cell Samples Undergoing Cell Death.
A polydisperse scattering model adapted for concentrated medium, namely the polydisperse structure factor model, was examined to explain the backscatter coefficients (BSCs) measured from packed cell samples undergoing cell death. Cell samples were scanned using high-frequency ultrasound in the 10-42 MHz bandwidth. A parameter estimation procedure was proposed to estimate the volume fraction and the relative impedance contrast that could explain the changes in BSC pattern by considering the actual change in cellular size distribution. Quantitative ultrasound parameters were estimated and related to the percentage of dead cells determined by flow cytometry. The standard deviation of scatterer size distribution extracted from the polydisperse structure factor model and the spectral intercept were found to be strongly correlated to the percentage of dead cells (r2 = 0.79 and r2 = 0.72, respectively). This study contributes to the understanding of ultrasonic scattering from cells undergoing cell death toward the monitoring of cancer therapy.


The effect of lipoprotein-associated phospholipase A2 deficiency on pulmonary allergic responses in Aspergillus fumigatus sensitized mice.
Lipoprotein-associated phospholipase A2 (Lp-PLA2)/platelet-activating factor acetylhydrolase (PAF-AH) has been implicated in the pathogenesis of cardiovascular disease. A therapeutic targeting of this enzyme was challenged by the concern that increased circulating platelet activating factor (PAF) may predispose to or increase the severity of the allergic airway response. The aim of this study was to investigate whether Lp-PLA2 gene deficiency increases the risk of PAF and IgE-mediated inflammatory responses in vitro and in vivo using mouse models. Lp-PLA2-/- mice were generated and back crossed to the C57BL/6 background. PAF-AH activity was measured using a hydrolysis assay in serum and bronchoalveolar lavage (BAL) samples obtained from mice. Aspergillus fumigatus (Af)-specific serum was prepared for passive allergic sensitization of mice in vivo and mast cells in vitro. β- hexosaminidase release was studied in bone marrow derived mast cells sensitized with Af-specific serum or DNP-IgE and challenged with Af or DNP, respectively. Mice were treated with lipopolysaccharide (LPS) and PAF intratracheally and studied 24 hours later. Mice were sensitized either passively or actively against Af and were studied 48 hours after a single intranasal Af challenge. Airway responsiveness to methacholine, inflammatory cell influx in the lung tissue and BAL, immunoglobulin (ELISA) and cytokine (Luminex) profiles were compared between the wild type (WT) and Lp-PLA2-/- mice. PAF-AH activity was reduced but not completely abolished in Lp-PLA2-/- serum or by in vitro treatment of serum samples with a high saturating concentration of the selective Lp-PLA2 inhibitor, SB-435495. PAF inhalation significantly enhanced airway inflammation of LPS treated WT and Lp-PLA2-/- mice to a similar extent. Sensitized WT and Lp-PLA2-/- bone-marrow derived mast cells released β-hexosaminidase following stimulation by allergen or IgE crosslinking to equivalent levels. Wild type and Lp-PLA2-/- mice responded to passive or active allergic sensitization by significant IgE production, airway inflammation and hyperresponsiveness after Af challenge. BAL cell influx was not different between these strains while IL-4, IL-5, IL-6 and eotaxin release was attenuated in Lp-PLA2-/- mice. There were no differences in the amount of total IgE levels in the Af sensitized WT and Lp-PLA2-/- mice. We conclude that Lp-PLA2 deficiency in C57BL/6 mice did not result in a heightened airway inflammation or hyperresponsiveness after PAF/LPS treatment or passive or active allergic sensitization and challenge.


Nurses and Twitter: the good, the bad, and the reluctant.
Nurses and other health professionals are adopting social media to network with health care professionals and organizations, support health education, deliver health promotion messages, enhance professional development and employment opportunities, and communicate within political forums. This paper explores the growing use of social media, and examines the current dynamics of Twitter as an example of the uptake of social media. This paper also offers practical guidance for new Twitter users who are interested in using this social media approach in clinical or educational settings, and for professional development.


A Gapless, Unambiguous Genome Sequence of the Enterohemorrhagic Escherichia coli O157:H7 Strain EDL933.
Escherichia coli EDL933 is the prototypic strain for enterohemorrhagic E. coli serotype O157:H7, associated with deadly food-borne outbreaks. Because the publicly available sequence of the EDL933 genome has gaps and >6,000 ambiguous base calls, we here present an updated high-quality, unambiguous genome sequence with no assembly gaps.


Persistence of transmitted HIV-1 drug resistance mutations associated with fitness costs and viral genetic backgrounds.
Transmission of drug-resistant pathogens presents an almost-universal challenge for fighting infectious diseases. Transmitted drug resistance mutations (TDRM) can persist in the absence of drugs for considerable time. It is generally believed that differential TDRM-persistence is caused, at least partially, by variations in TDRM-fitness-costs. However, in vivo epidemiological evidence for the impact of fitness costs on TDRM-persistence is rare. Here, we studied the persistence of TDRM in HIV-1 using longitudinally-sampled nucleotide sequences from the Swiss-HIV-Cohort-Study (SHCS). All treatment-naïve individuals with TDRM at baseline were included. Persistence of TDRM was quantified via reversion rates (RR) determined with interval-censored survival models. Fitness costs of TDRM were estimated in the genetic background in which they occurred using a previously published and validated machine-learning algorithm (based on in vitro replicative capacities) and were included in the survival models as explanatory variables. In 857 sequential samples from 168 treatment-naïve patients, 17 TDRM were analyzed. RR varied substantially and ranged from 174.0/100-person-years;CI=[51.4, 588.8] (for 184V) to 2.7/100-person-years;[0.7, 10.9] (for 215D). RR increased significantly with fitness cost (increase by 1.6[1.3,2.0] per standard deviation of fitness costs). When subdividing fitness costs into the average fitness cost of a given mutation and the deviation from the average fitness cost of a mutation in a given genetic background, we found that both components were significantly associated with reversion-rates. Our results show that the substantial variations of TDRM persistence in the absence of drugs are associated with fitness-cost differences both among mutations and among different genetic backgrounds for the same mutation.


Cross relaxation in nitroxide spin labels.
Cross relaxation, and mI-dependence of the intrinsic electron spin-lattice relaxation rate We, are incorporated explicitly into the rate equations for the electron-spin population differences that govern the saturation behaviour of 14N- and 15N-nitroxide spin labels. Both prove important in spin-label EPR and ELDOR, particularly for saturation recovery studies. Neither for saturation recovery, nor for CW-saturation EPR and CW-ELDOR, can cross relaxation be described simply by increasing the value of We, the intrinsic spin-lattice relaxation rate. Independence of the saturation recovery rates from the hyperfine line pumped or observed follows directly from solution of the rate equations including cross relaxation, even when the intrinsic spin-lattice relaxation rate We is mI-dependent.


Feeling anxious: a twin study of panic/somatic ratings, anxiety sensitivity and heartbeat perception in children.
Little is known about mechanisms of genetic influence on panic, particularly in childhood. Cognitive theories of panic disorder highlight threatening interpretations of physical sensations, and increased awareness of such sensations. Specifically, anxiety sensitivity (AS) and heartbeat perception (HBP) have been associated with panic in adults and children. We examined genetic and environmental influences on childhood AS, HBP, panic/somatic ratings, and their associations. Self-ratings of AS and DSM-based anxiety (including panic/somatic items) were obtained from 300 eight-year-old twin pairs (600 individuals), selected for mother-rated child anxiety at age 7. HBP was also assessed. Panic/somatic ratings were significantly correlated with both AS (r = .55) and continuous HBP error scores (r = -.13). AS and HBP scores showed significantly greater correlations with panic/somatic ratings than with all other anxiety scales, except for HBP and school anxiety. Genetic influences on panic/somatic ratings were modest (15%), and moderate for both AS (37%), and HBP (30%). Non-shared environmental influences were substantial. The genetic correlations between panic/somatic ratings and both AS and HBP error scores were .98 (95% CI: .74-1.00) and -.46 (95% CI: -1.00-1.00) respectively. Self-ratings of panic and AS overlap genetically. Future research should consider whether AS mediates genetic risk for panic disorder.


Property Decoupling across the Embryonic Nucleus-Melt Interface during Polymer Crystal Nucleation.
Spatial distributions are presented that quantitatively capture how polymer properties (e.g., segment alignment, density, and potential energy) vary with distance from nascent polymer crystals (nuclei) in prototypical polyethylene melts. It is revealed that the spatial extent of nuclei and their interfaces is metric-dependent as is the extent to which nucleus interiors are solid-like. As distance from a nucleus increases, some properties, such as density, decay to melt-like behavior more rapidly than polymer segment alignment, indicating that a polymer nucleus resides in a nematic-like droplet. This nematic-like droplet region coincides with enhanced formation of ordered polymer segments that are not part of the nucleus. It is more favorable to find nonconstituent ordered polymer segments near a nucleus than in the surrounding metastable melt, pointing to the possibility of one nucleus inducing the formation of other nuclei. In this vein, there is also a second region of enhanced ordering that lies along the nematic director of a nucleus, but beyond its nematic droplet and fold regions. These results indicate that crystal stacking, a key characteristic of lamellae in semicrystalline polymeric materials, begins to emerge during the earliest stages of polymer crystallization (i.e., crystal nucleation). More generally, the findings of this study provide a conceptual bridge between polymer crystal nucleation under nonflow and flow conditions and are used to rationalize previous results.


Formation and role of exosomes in cancer.
Exosomes offer new insight into cancer biology with both diagnostic and therapeutic implications. Because of their cell-to-cell communication, exosomes influence tumor progression, metastasis, and therapeutic efficacy. They can be isolated from blood and other bodily fluids to reveal disease processes occurring within the body, including cancerous growth. In addition to being a reservoir of cancer biomarkers, they can be re-engineered to reinstate tumor immunity. Tumor exosomes interact with various cells of the microenvironment to confer tumor-advantageous changes that are responsible for stromal activation, induction of the angiogenic switch, increased vascular permeability, and immune escape. Exosomes also contribute to metastasis by aiding in the epithelial-to-mesenchymal transition and formation of the pre-metastatic niche. Furthermore, exosomes protect tumor cells from the cytotoxic effects of chemotherapy drugs and transfer chemoresistance properties to nearby cells. Thus, exosomes are essential to many lethal elements of cancer and it is important to understand their biogenesis and role in cancer.


Cryptochrome-dependent and -independent circadian entrainment circuits in Drosophila.
Entrainment to environmental light/dark (LD) cycles is a central function of circadian clocks. In Drosophila, entrainment is achieved by Cryptochrome (CRY) and input from the visual system. During activation by brief light pulses, CRY triggers the degradation of TIMELESS and subsequent shift in circadian phase. This is less important for LD entrainment, leading to questions regarding light input circuits and mechanisms from the visual system. Recent studies show that different subsets of brain pacemaker clock neurons, the morning (M) and evening (E) oscillators, have distinct functions in light entrainment. However, the role of CRY in M and E oscillators for entrainment to LD cycles is unknown. Here, we address this question by selectively expressing CRY in different subsets of clock neurons in a cry-null (cry(0)) mutant background. We were able to rescue the light entrainment deficits of cry(0) mutants by expressing CRY in E oscillators but not in any other clock neurons. Par domain protein 1 molecular oscillations in the E, but not M, cells of cry(0) mutants still responded to the LD phase delay. This residual light response was stemming from the visual system because it disappeared when all external photoreceptors were ablated genetically. We concluded that the E oscillators are the targets of light input via CRY and the visual system and are required for normal light entrainment.


Shared Genetic Factors Underlie Migraine and Depression.
Migraine frequently co-occurs with depression. Using a large sample of Australian twin pairs, we aimed to characterize the extent to which shared genetic factors underlie these two disorders. Migraine was classified using three diagnostic measures, including self-reported migraine, the ID migraine™ screening tool, or migraine without aura (MO) and migraine with aura (MA) based on International Headache Society (IHS) diagnostic criteria. Major depressive disorder (MDD) and minor depressive disorder (MiDD) were classified using the Diagnostic and Statistical Manual of Mental Disorders (DSM) criteria. Univariate and bivariate twin models, with and without sex-limitation, were constructed to estimate the univariate and bivariate variance components and genetic correlation for migraine and depression. The univariate heritability of broad migraine (self-reported, ID migraine, or IHS MO/MA) and broad depression (MiDD or MDD) was estimated at 56% (95% confidence interval [CI]: 53-60%) and 42% (95% CI: 37-46%), respectively. A significant additive genetic correlation (r G = 0.36, 95% CI: 0.29-0.43) and bivariate heritability (h 2 = 5.5%, 95% CI: 3.6-7.8%) was observed between broad migraine and depression using the bivariate Cholesky model. Notably, both the bivariate h 2 (13.3%, 95% CI: 7.0-24.5%) and r G (0.51, 95% CI: 0.37-0.69) estimates significantly increased when analyzing the more narrow clinically accepted diagnoses of IHS MO/MA and MDD. Our results indicate that for both broad and narrow definitions, the observed comorbidity between migraine and depression can be explained almost entirely by shared underlying genetically determined disease mechanisms.


Differential requirement for beta-catenin in epithelial and fiber cells during lens development.
Recent studies implicate Wnt/beta-catenin signaling in lens differentiation (Stump, R. J., et al., 2003. A role for Wnt/beta-catenin signaling in lens epithelial differentiation. Dev Biol;259:48-61). Beta-catenin is a component of adherens junctions and functions as a transcriptional activator in canonical Wnt signaling. We investigated the effects of Cre/LoxP-mediated deletion of beta-catenin during lens development using two Cre lines that specifically deleted beta-catenin in whole lens or only in differentiated fibers, from E13.5. We found that beta-catenin was required in lens epithelium and during early fiber differentiation but appeared to be redundant in differentiated fiber cells. Complete loss of beta-catenin resulted in an abnormal and deficient epithelial layer with loss of E-cadherin and Pax6 expression as well as abnormal expression of c-Maf and p57(kip2) but not Prox1. There was also disrupted fiber cell differentiation, characterized by poor cell elongation, decreased beta-crystallin expression, epithelial cell cycle arrest at G(1)-S transition and premature cell cycle exit. Despite cell cycle arrest there was no induction of apoptosis. Mutant fiber cells displayed altered apical-basal polarity as evidenced by altered distribution of the tight junction protein, ZO1, disruption of apical actin filaments and abnormal deposition of extracellular matrix, resulting in a deficient lens capsule. Loss of beta-catenin also affected the formation of adhesion junctions as evidenced by dissociation of N-cadherin and F-actin localization in differentiating fiber cells. However, loss of beta-catenin from terminally differentiating fibers had no apparent effects on adhesion junctions between adjacent embryonic fibers. These data indicate that beta-catenin plays distinct functions during lens fiber differentiation and is involved in both Wnt signaling and adhesion-related mechanisms that regulate lens epithelium and early fiber differentiation.


Dentin permeability after toothbrushing with different toothpastes.
To evaluate the interaction of smear layer produced during brushing and dentifrice particles. Dentin disks were obtained from extracted human third molars. Dentin permeability was evaluated using a hydraulic pressure apparatus working at 1 psi of pressure. After preparation, each sample was connected to the hydraulic pressure apparatus to evaluate the permeability of dentin with the smear layer produced during specimen preparation. After 5 min of measurements dentin was etched with 37% phosphoric acid gel for 2 min, washed and gently dried with cotton to measure the permeability after smear layer removal. This was done to measure the maximum permeability of each specimen (expressed as 100%). Then a second smear layer was produced using a #400 carbide paper under water for 1 min. Dentin permeability of the smear layer covered dentin was then measured, and expressed as a percent of the maximum permeability of that specimen, permitting each specimen to serve as its own control. The three toothpastes used (Merfluan powder, Merfluan paste and Fluorigard) were applied on dentin surfaces using a small sponge to completely cover the dentin surface. After 5 s, each sample was connected to a mechanical device and brushed under water for 3 min with constant pressure of 250 gr using a Colgate medium toothbrush. After this treatment, each sample was gently washed with tap water, gently air dried for 3-5 s and connected with the pressure apparatus to remeasure the permeability after brushing. For SEM examination of dentin before and after treatment with toothpaste and brushing, each dentin and enamel sample was fixed in 2% buffered glutaraldehyde. Dentin permeability proved unaffected by dentifrice treatments. SEM observations demonstrated the presence of dentifrice particles on dentin surface and inside dentin tubules. Smear plugs produced during dentin brushing were not removed by dentifrices.


Scalable Image Coding Based on Epitomes.
In this paper, we propose a novel scheme for scalable image coding based on the concept of epitome. An epitome can be seen as a factorized representation of an image. Focusing on spatial scalability, the enhancement layer of the proposed scheme contains only the epitome of the input image. The pixels of the enhancement layer not contained in the epitome are then restored using two approaches inspired from local learning-based super-resolution methods. In the first method, a locally linear embedding model is learned on base layer patches and then applied to the corresponding epitome patches to reconstruct the enhancement layer. The second approach learns linear mappings between pairs of co-located base layer and epitome patches. Experiments have shown that the significant improvement of the rate-distortion performances can be achieved compared with the Scalable extension of HEVC (SHVC).


Molecular cloning of complementary DNA from a pneumopathic strain of bovine viral diarrhea virus and its diagnostic application.
A pneumopathic strain of bovine viral diarrhea virus was grown in cell culture and purified. Genomic ribonucleic acid was extracted, polyadenylated at the 3' end, and copied into complementary DNA after oligo-dT priming. Complementary DNA was male double stranded and cloned into the pUC9 plasmid. Approximately 200 complementary DNA clones varying in length from 0.5 to 2.5 kilobases were obtained. Hybridization assays indicated that the sequences isolated were specific for bovine viral diarrhea virus and that at least 5.5 kilobases of bovine viral diarrhea virus genome was represented in the library of complementary DNA clones, the majority of which may have originated from the 3' end of the virus genome. One cloned complementary DNA sequence was used as a 32P-labelled hybridization probe for bovine viral diarrhea virus detection. The probe hybridized with all cytopathic and noncytopathic strains of bovine viral diarrhea virus tested and was 100 times more sensitive than infectivity assays for the detection of bovine viral diarrhea virus. Hybridization did not occur with nucleic acids from bovine coronavirus, bluetongue virus, bovine adenovirus or uninfected cell cultures. Native plasmid DNA sequences, labelled with 32P, did not hybridize with bovine viral diarrhea virus ribonucleic acid.


The relationship of bleeding patterns to daily reproductive hormones in women approaching menopause.
To discover early hormonal predictors of menopause and the stages of the menopausal transition and to understand the hormonal basis behind the bleeding abnormalities common in the menopausal transition. A cohort of 804 women aged 42-52 collected first void urine samples daily for one complete menstrual cycle or 50 days (whichever came first) once a year for 3 years. Urine was assayed for excreted levels of follicle-stimulating hormone, luteinizing hormone, estrogen metabolites, and progesterone metabolites, which were normalized for creatinine concentration. Anovulation was defined by an algorithm based on progesterone secretion. Menstrual bleeding parameters were derived from daily calendars. Correlations among bleeding characteristics, hormone concentrations, and other potential clinical predictors were analyzed using multivariable logistic regression models. An ethnically diverse population of women (mean age of 47) with a majority in the early perimenopause was studied. Approximately 20% of all cycles were anovulatory. Short cycle intervals (fewer than 21 days) were common early in the menopause transition and were associated with anovulation (44%). Long cycle intervals (more than 36 days) also were associated with anovulatory cycles (65%). Both short (1-3 days) and long (more than 8 days) duration of menstrual bleeding were associated with anovulation (18% and 23%, respectively). Women with anovulatory cycles were less likely to report heavy menstrual bleeding as compared with those with ovulatory cycles. Heavy bleeding was not associated with steroid hormone concentrations but was associated with obesity and with the self-reported presence of leiomyomata. Among women in the early menopause transition, abnormalities in timing of menstrual bleeding (cycle intervals or bleeding duration) have a hormonal basis and are frequently associated with anovulation. In contrast, abnormally heavy periods do not appear to have a steroid hormonal basis and are less likely after anovulatory cycles. Heavy periods are associated with obesity and leiomyomata. II.


Sex differences in parking are affected by biological and social factors.
The stereotype of women's limited parking skills is deeply anchored in modern culture. Although laboratory tests prove men's average superiority in visuospatial tasks and parking requires complex, spatial skills, underlying mechanisms remain unexplored. Here, we investigated performance of beginners (nine women, eight men) and more experienced drivers (21 women, 27 men) at different parking manoeuvres. Furthermore, subjects conducted the mental rotation test and self-assessed their parking skills. We show that men park more accurately and especially faster than women. Performance is related to mental rotation skills and self-assessment in beginners, but only to self-assessment in more experienced drivers. We assume that, due to differential feedback, self-assessment incrementally replaces the controlling influence of mental rotation, as parking is trained with increasing experience. Results suggest that sex differences in spatial cognition persist in real-life situations, but that socio-psychological factors modulate the biological causes of sex differences.


Accelerated skeletal muscle recovery after in vivo polyphenol administration.
Acute skeletal muscle damage results in fiber disruption, oxidative stress and inflammation. We investigated cell-specific contributions to the regeneration process after contusion-induced damage (rat gastrocnemius muscle) with or without chronic grape seed-derived proanthocyanidolic oligomer (PCO) administration. In this placebo-controlled study, male Wistar rats were subjected to PCO administration for 2 weeks, after which they were subjected to a standardised contusion injury. Supplementation was continued after injury. Immune and satellite cell responses were assessed, as well as oxygen radical absorption capacity and muscle regeneration. PCO administration resulted in a rapid satellite cell response with an earlier peak in activation (Pax7⁺, CD56⁺, at 4 h post-contusion) vs. placebo groups (PLA) (P<.001: CD56⁺ on Day 5 and Pax7⁺ on Day 7). Specific immune-cell responses in PLA followed expected time courses (neutrophil elevation on Day 1; sustained macrophage elevation from Days 3 to 5). PCO dramatically decreased neutrophil elevation to nonsignificant, while macrophage responses were normal in extent, but significantly earlier (peak between Days 1 and 3) and completely resolved by Day 5. Anti-inflammatory cytokine, IL-10, increased significantly only in PCO (Day 3). Muscle fiber regeneration (MHC(f) content and central nuclei) started earlier and was complete by Day 14 in PCO, but not in PLA. Thus, responses by three crucial cell types involved in muscle recovery were affected by in vivo administration of a specific purified polyphenol in magnitude (neutrophil), time course (macrophages), or time course and activation state (satellite cell), explaining faster effective regeneration in the presence of proanthocyanidolic oligomers.


Effectiveness of educational interventions to improve food safety practices among older adults.
The purpose of the study was to develop and evaluate the effectiveness of using Web-based and print materials for improving food safety practices to reduce the risk of foodborne illness among older adults. The study used a randomized controlled design, with participants assigned to an intervention group or control group. Although we observed small improvements in both groups, the difference in the changes between the two groups was nonsignificant, suggesting the educational materials did not impact participant behavior. We did, however, observe a trend improvement in one measure: the recommendation to avoid eating cold (not reheated) deli meats. The lack of program impact may be attributable to limitations of the evaluation (e.g., measurement effects) or the intervention (e.g., lack of personal contact). Based on the survey findings, improvements in older adults' food safety practices regarding reheating deli meats to steaming hot and cooking eggs until the yolks and whites are firm are needed. The current study and previous research suggest that current cohorts of older adults may be more receptive to print materials than Web-based materials. To improve retention and adoption of recommended food safety practices among older adults, future educational interventions should focus on a limited number of practices and combine print materials with personal contact.


Development and validation of the Social Reward Questionnaire-Early Childhood.
Social interactions like group inclusion, receiving praise, or treating others kindly can be motivating and enjoyable. Social reward sensitivity, including motivation and enjoyment, varies between individuals. In early childhood, this variation may relate to differences in social experience and development. Social reward questionnaires have been developed to measure individual differences in social enjoyment for adolescents and adults, but no early childhood measure currently exists. Here, we describe the development and validation of the parent/caregiver report Social Reward Questionnaire-Early Childhood (SRQ-EC) for children aged 3-7 years. The SRQ-EC was developed to quantify both wanting (motivation) and liking (enjoyment) of social rewards, which were considered in separate factor models. For wanting and liking models, exploratory (N = 126) and confirmatory (N = 344) factor analyses identified that three subscales best represented early childhood social reward sensitivity, which were: Sociability (large groups), Admiration (praise and positive attention), and Prosocial Interactions and Compliance (kindness and rule following). SRQ-EC subscales were internally consistent (ω = 0.76-0.91, α = 0.75-0.88, mean interitem correlations = 0.38-0.60) with high test-retest reliability over 2-weeks (r = 0.66-0.85, all p < .001). Subscales differentially associated with other social behavior and personality measures, suggesting construct validity. SRQ-EC subscale scores further showed differential and significant associations with autistic-like traits in nonautistic children. These results suggest that SRQ-EC subscale scores are reliable for assessing social reward sensitivity during early childhood, which could offer key developmental insight regarding interindividual variation in early social behavior. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Determinants of cluster size in large, population-based molecular epidemiology study of tuberculosis, northern Malawi.
Tuberculosis patients with identical strains of Mycobacterium tuberculosis are described as clustered. Cluster size may depend on patient or strain characteristics. In a 7-year population-based study of tuberculosis in Karonga District, Malawi, clusters were defined by using IS6110 restriction fragment length polymorphism, excluding patterns with <5 bands. Spoligotyping was used to compare strains with an international database. Among 682 clustered patients, cluster size ranged from 2 to 37. Male patients, young adults, and town residents were over-represented in large clusters. Cluster size was not associated with HIV status or death from tuberculosis. Spoligotypes from 9 (90%) of 10 large cluster strains were identical or very similar (1 spacer different) to common spoligotypes found elsewhere, compared with 37 (66%) of 56 of those from nonclustered patients (p = 0.3). Large clusters were associated with factors likely to be related to social mixing, but spoligotypes of common strains in this setting were also common types elsewhere, consistent with strain differences in transmissibility.


Sclerotinia Rot of Windflower (Pulsatilla koreana Nakai) Caused by Sclerotinia nivalis in China.
Windflowers (Pulsatilla spp.) are perennial medicinal plants in the family Ranunculaceae with high economic as well as medicinal value in China. It is a commonly used traditional Chinese medicine (1). In 2012, a stem rot disease was observed on windflower (P. koreana Nakai) at flowering stages in fields of Liaoning Province, China. Disease incidence ranged from 10 to 65% (average of 45%) and resulted in approximately 25 to 60% yield loss. The infected area of plants initially takes on a dark green or brown water-soaked appearance and then becomes paler. Soon after, plants turn brown or black and die. If moisture conditions remain conducive, a cottony mycelium cover was observed on the affected area. Later, the mycelium usually produces numerous black sclerotia that were up to 1 cm long in affected plant parts. Diseased tissue were surface sterilized for 1 min in 1% NaOCl (v/v), rinsed with sterilized distilled water, and plated on potato dextrose agar (PDA). Isolates were mesophilic, with an optimum growth temperature of around 20°C. Colonies on PDA were white, with abundant aerial mycelium. Some mycelium in the colony center, especially that submerged in the medium, was light brown. Sclerotia were spherical to subspherical, elongated or fused to form irregular shapes, 2.5 to 9.0 × 2.0 to 6.8 mm. They were tightly attached to the agar surface by their under surface, which could be seen through the bottom of the petri dishes. These characteristics were consistent with Sclerotinia nivalis (2,3). Two isolates were selected for molecular identification. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (4) and sequenced. The obtained sequences (GenBank Accession Nos. JX206828 and JX424615) showed 99 and 100% homology with the sequences of S. nivalis in GenBank (Accession No. AB516670). To demonstrate pathogenicity, mycelial blocks of the isolates grown on PDA were placed on the base of the stems of ten 1-month-old Pulsatilla koreana plants. The same numbers of control plants were treated with PDA plugs as a control. The inoculated plants were incubated at 25°C with a 12-h photoperiod. After 4 days, the initiation of stem necrosis was observed, and 9 days after inoculation, the plants collapsed and died. Control plants had no symptoms. The same fungus was reisolated from all inoculated plants, satisfying Koch's postulates. To our knowledge, this is the first confirmed report of windflower as a natural host for S. nivalis in China. References: (1) S. C. Bang et al. J. Nat. Prod. 68:268, 2005. (2) G. Q. Li et al. Mycol. Res. 104:232, 2000. (3) I. Saito. Mycoscience. 38:227, 1997. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.


Allosteric regulators of the proteasome: potential drugs and a novel approach for drug design.
The proteasome recently gained an exceptional attention as a novel drug target, therefore its inhibitors became important subjects for rational drug design. A synthetic competitive inhibitor Velcade was lately approved in a fast-track process to treat multiple myeloma and is tested with other types of cancers. The proteasome is a major proteolytic assembly in eukaryotic cells responsible for the degradation of most intracellular proteins, including proteins crucial to cell cycle regulation and apoptosis. The ubiquitin-proteasome pathway has been implicated in many diseases such as cancer, autoimmune diseases, inflammation, and stroke. The activity of the proteasome can be blocked for therapeutic purposes with competitive inhibitors like Velcade, which trigger apoptosis in target cells. However, much more versatile outcomes and a true control of the proteasome can be achieved with allosteric regulators. Certain natural proteins and peptides bind to the catalytic core of the proteasome and allosterically induce a wide array of effects ranging from changes in product size to substrate-specific inhibition. Designing small synthetic compounds allosterically interacting with the proteasome represents a novel approach that has enormous potential for the treatment of a wide range of diseases. Below we provide a review of current knowledge about proteasomal allosteric ligands.


Daclatasvir: the first of a new class of drugs targeted against hepatitis C virus NS5A.
Hepatitis C virus (HCV) infection affects about 160 million people worldwide. It is treated with pegylatedinterferon (peg-IFN) and ribavirin, and in the case of patients affected by genotype 1, also with a protease inhibitor (telaprevir or boceprevir). Despite a good success rate, IFN-based combinations are contraindicated in several patients (e.g. decompensated cirrhosis, patients with psychiatric disorders, severe heart diseases or autoimmune disorders) and are associated with frequent adverse events that ultimately reduce their use. Numerous oral drugs are in an advanced phase of clinical development, and in some cases, in IFN-free combinations. This review focuses on preclinical and clinical data regarding daclatasvir (BMS-790052), which is a highly selective HCV NS5A replication complex inhibitor effective against HCV genotypes 1, 2, 3 and 4. In vitro data show that daclatasvir exerts a very potent antiviral effect against several HCV genotypes. Its pharmacokinetics is optimal and allows once-a-day oral administration. Its adverse event profile is good. Clinical data regarding its efficacy in combination with peg-IFN, ribavirin or other direct antiviral agents are impressive (rates of sustained virological response range between 60% and 100% in treatment-naïve patients). The only drawback of this drug appears to be a relatively low genetic barrier to resistance. In conclusion, daclatasvir, especially in combinations with other antiviral agents, is a very promising drug for the treatment of chronic hepatitis C.


Powerful and tunable operation of a 1-2-kHz repetition-rate gain-switched Cr:forsterite laser and its frequency doubling.
We present a comprehensive study of the optimum operating regime in gain-switched Cr:forsterite lasers pumped at kilohertz repetition rates, comparing five crystals of similar quality but different dopant levels. The optimization of the cavity design includes selection of the proper pump fluence to account for excited-state absorption, optimum matching of the pump and laser modes, and consideration of thermal effects. As a result >1-W average output power is demonstrated at 2 kHz. The maximum conversion efficiencies achieved at 1 kHz are 24.2% (slope) and 20% (absolute). Narrow-band operation of this laser is possible with a birefringent filter, which is a prerequisite for efficient frequency doubling to cover the 585-660-nm part of the visible spectral range. Tunable second-harmonic generation in a temperature-tuned noncritical scheme that employs LiB(3)O(5) produces 60 mW of average power near 619 nm with 13.5% conversion efficiency.


Temperament and character in eating disorders: ten years of studies.
In recent years a number of studies of personality have been performed in subjects with Eating Disorders (EDs) to investigate the clinical differences between controls and ED patients and among EDs subtypes, and its role in the development and course of symptoms. The Tridimensional Personality Questionnaire (TPQ) and the Temperament and Character Inventory (TCI) have been widely used at this purpose, allowing the description of specific temperament and character profiles for EDs. High Harm Avoidance (HA) and low Self-Directedness (SD) are shared by all EDs. Slight differences on some facets have been found among ED subgroups. Nevertheless, HA is influenced by mood and both high HA and low SD are personality traits shared by many mental disorders, whose specificity is rather low. Restrictor anorectics are characterized by high Persistence (P) and a relatively higher SD, and bulimics by higher Novelty Seeking (NS) and the lowest SD, while binge/purging and purging anorectics share some traits with anorexia and some with bulimia. Though current data justify the discrimination among anorexia subtypes, they are not in contrast with the thesis of a continuum in ED personality traits. Since some personality traits display a prognostic value with regard to therapy and clinical outcome, further studies are needed on treatments and prognostic factors in EDs. Moreover, studies attempting to define the neurobiological and genetic correlates of temperament should be supported by clinical pharmacological trials.


Decreased benzodiazepine receptor binding in prefrontal cortex in combat-related posttraumatic stress disorder.
Animals exposed to stress exhibit a decrease in benzodiazepine receptor binding in the frontal cortex. No studies have examined central benzodiazepine receptor binding in patients with posttraumatic stress disorder (PTSD). The purpose of this study was to examine measures of benzodiazepine receptor binding in PTSD. From 13 patients with Vietnam combat-related PTSD and 13 case-matched healthy comparison subjects, a quantitative measure related to benzodiazepine receptor binding (distribution volume) was obtained with single photon emission computed tomography (SPECT) imaging of [(123)I]iomazenil binding and measurement of radioligand concentration in plasma. Distribution volume image data were analyzed by means of statistical parametric mapping. Lower distribution volumes were found in the prefrontal cortex (Brodmann's area 9) of PTSD patients than in comparison subjects. These findings of lower values for the benzodiazepine receptor binding measure of distribution volume are consistent with fewer benzodiazepine receptors and/or reduced affinity of receptor binding in the medial prefrontal cortex in patients with PTSD. Alterations in benzodiazepine receptor function in this area may underlie many of the symptoms of PTSD.


A novel tri-enzyme system in combination with laser-driven NMR enables efficient nuclear polarization of biomolecules in solution.
NMR is an extremely powerful, yet insensitive technique. Many available nuclear polarization methods that address sensitivity are not directly applicable to low-concentration biomolecules in liquids and are often too invasive. Photochemically induced dynamic nuclear polarization (photo-CIDNP) is no exception. It needs high-power laser irradiation, which often leads to sample degradation, and photosensitizer reduction. Here, we introduce a novel tri-enzyme system that significantly overcomes the above challenges, rendering photo-CIDNP a practically applicable technique for NMR sensitivity enhancement in solution. The specificity of the nitrate reductase (NR) enzyme is exploited to selectively in situ reoxidize the reduced photo-CIDNP dye FMNH2. At the same time, the oxygen-scavenging ability of glucose oxidase (GO) and catalase (CAT) is synergistically employed to prevent sample photodegradation. The resulting tri-enzyme system (NR-GO-CAT) enables prolonged sensitivity-enhanced data collection in 1D and 2D heteronuclear NMR, leading to the highest photo-CIDNP sensitivity enhancement (48-fold relative to SE-HSQC) achieved to date for amino acids and polypeptides in solution. NR-GO-CAT extends the concentration limit of photo-CIDNP NMR down to the low micromolar range. In addition, sensitivity (relative to the reference SE-HSQC) is found to be inversely proportional to sample concentration, paving the way for the future analysis of even more diluted samples.


The effects of acute L-carnitine supplementation on endurance performance of athletes.
This study examined the effect of acute L-carnitine loading on the endurance performance of footballers. Measurements were performed on 26 candidate professional footballers who volunteered to take part in the study. Athletes were given a glass of fruit juice 1 hour before applying L-carnitine with the double-blind method. Then, 12 participants were given 3 g of L-carnitine (LK-3) and the remaining 14 were given 4 g (LK-4). Athletes began the exercise test at a running speed of 8 km·h and then continued at 10 km·h. The speed was increased 1 km·h every 3 minutes, and the test continued until the subject chose to quit. Heart rate was registered using a portable telemetric heart rate monitor during the test. Blood samples were taken from the earlobes of the footballers both before the test and before the speed increase (during the 1-minute interval), and the lactate (La) concentration was measured electroenzymatically. The test was repeated after 1 week as a group of placebos (P-3 and P-4). The result showed that the running speeds corresponding to specific La concentrations were increased, and La and heart rate responses to the running speeds were decreased in both supplemented groups compared with placebos (p ≤ 0.05). A significant reduction in heart rate was found in LK-4 and P-4 (p ≤ 0.05). When the Borg responses to the running speeds were analyzed, a significant difference was found in both supplemented groups (p ≤ 0.05). The results show that 3 or 4 g of L-carnitine taken before physical exercise prolonged exhaustion.


Beneficial effects of potassium on human health.
Until recently, humans consumed a diet high in potassium. However, with the increasing consumption of processed food, which has potassium removed, combined with a reduction in the consumption of fruits and vegetables, there has been a large decrease in potassium intake which now, in most developed countries, averages around 70 mmol day-1, i.e. only one third of our evolutionary intake. Much evidence shows that increasing potassium intake has beneficial effects on human health. Epidemiological and clinical studies show that a high-potassium diet lowers blood pressure in individuals with both raised blood pressure and average population blood pressure. Prospective cohort studies and outcome trials show that increasing potassium intake reduces cardiovascular disease mortality. This is mainly attributable to the blood pressure-lowering effect and may also be partially because of the direct effects of potassium on the cardiovascular system. A high-potassium diet may also prevent or at least slow the progression of renal disease. An increased potassium intake lowers urinary calcium excretion and plays an important role in the management of hypercalciuria and kidney stones and is likely to decrease the risk of osteoporosis. Low serum potassium is strongly related to glucose intolerance, and increasing potassium intake may prevent the development of diabetes that occurs with prolonged treatment with thiazide diuretics. Reduced serum potassium increases the risk of lethal ventricular arrhythmias in patients with ischaemic heart disease, heart failure and left ventricular hypertrophy, and increasing potassium intake may prevent this. The best way to increase potassium intake is to increase the consumption of fruits and vegetables.


DNA methylation at the Igf2/H19 imprinting control region is associated with cerebellum mass in outbred mice.
Insulin-like growth factor 2 (Igf2) is a paternally expressed imprinted gene regulating fetal growth, playing an integral role in the development of many tissues including the brain. The parent-of-origin specific expression of Igf2 is largely controlled by allele-specific DNA methylation at CTCF-binding sites in the imprinting control region (ICR), located immediately upstream of the neighboring H19 gene. Previously we reported evidence of a negative correlation between DNA methylation in this region and cerebellum weight in humans. We quantified cerebellar DNA methylation across all four CTCF binding sites spanning the murine Igf2/H19 ICR in an outbred population of Heterogeneous Stock (HS) mice (n = 48). We observe that DNA methylation at the second and third CTCF binding sites in the Igf2/H19 ICR shows a negative relationship with cerebellar mass, reflecting the association observed in human post-mortem cerebellum tissue. Given the important role of the cerebellum in motor control and cognition, and the link between structural cerebellar abnormalities and neuropsychiatric phenotypes, the identification of epigenetic factors associated with cerebellum growth and development may provide important insights about the etiology of psychiatric disorders.


Ensembl 2013.
The Ensembl project (http://www.ensembl.org) provides genome information for sequenced chordate genomes with a particular focus on human, mouse, zebrafish and rat. Our resources include evidenced-based gene sets for all supported species; large-scale whole genome multiple species alignments across vertebrates and clade-specific alignments for eutherian mammals, primates, birds and fish; variation data resources for 17 species and regulation annotations based on ENCODE and other data sets. Ensembl data are accessible through the genome browser at http://www.ensembl.org and through other tools and programmatic interfaces.


"Black" responses dominate macaque primary visual cortex v1.
Achromatic visual information is transferred from the retina to the brain through two parallel channels: ON-center cells carry "white" information and OFF-center cells "black" information (Nelson et al., 1978; Schiller, 1982; Schiller et al., 1986). Responses of ON and OFF retinal and thalamic neurons are approximately equal in magnitude (Krüger and Fischer, 1975; Kremers et al., 1993), but psychophysical studies have shown that humans detect light decrements (black) better and faster than increments (white) (Blackwell, 1946; Short, 1966; Krauskopf, 1980; Whittle, 1986; Bowen et al., 1989; Chan and Tyler, 1992; Kontsevich and Tyler, 1999; Chubb and Nam, 2000; Dannemiller and Stephens, 2001). From recordings of single-cell activity in the macaque monkey's primary visual cortex (V1), we found that black-dominant neurons substantially outnumbered white-dominant neurons in the corticocortical output layers 2/3, but the numbers of black- and white-dominant neurons were nearly equal in the thalamocortical input layer 4c. These results strongly suggest that the black-over-white preference is generated or greatly amplified in V1. The predominance of OFF neurons in layers 2/3 of V1, which provide visual input to higher cortical areas, may explain why human subjects detect black more easily than white. Furthermore, our results agree with human EEG and fMRI findings that V1 responses to decrements are stronger than to increments, though the OFF/ON imbalance we found in layers 2/3 of macaque V1 is much larger than in the whole V1 population in the human V1 experiments (Zemon et al., 1988, 1995; Olman et al., 2008).


The pattern of femoral diaphyseal fractures in children admitted in Sarawak General Hospital.
Trend towards changing the face of management for pediatric femoral fractures tends to advocate operative treatment. This study was undertaken to review our current practice in the wake of recent progress in the management of pediatric femoral fractures. Fifty patients with femoral diaphyseal fracture treated in Sarawak General Hospital were reviewed retrospectively after an average follow-up of 2.6 years. There were 36 boys and 14 girls, with a mean age of 6.2 years (range five months to 14 years). Children under six years of age constituted the majority of the patients. Half of the fractures were caused by road traffic accident. Nine patients had associated injuries. The most common site of fracture was at the middle third (N=31). The treatment regimens were delayed hip spica (DHS) in 16, immediate hip spica (IHS) in 24, plate osteosynthesis (PO) in five, titanium elastic nailing (TEN) in five, and external fixation (EF) in one. The minimum hospital stay was two days, and the maximum 33 days (mean, 9.7 days). Malunion was the commonest complication. Conservative treatment is the preferred option for children under six years of age. It is cost-effective with minimal complication. The other treatment options are reserved for specific indication in older children. Diaphyseal fractures of the femur in children can be adequately managed non-operatively.


Breast Cancer Therapy-Related Cardiac Dysfunction in Adult Women Treated in Routine Clinical Practice: A Population-Based Cohort Study.
Most women diagnosed with breast cancer are younger than 65 years of age. Population-based studies on cancer therapy-related cardiotoxicity have focused on older women. We sought to determine the risk of cardiotoxicity with breast cancer therapy in women with an age distribution representative of routine clinical practice. This was a population-based retrospective cohort study including 14 regional cancer centers in Ontario, Canada. Adult women receiving chemotherapy for stage I to III breast cancer between 2007 and 2012 were included. Cancer treatment was categorized as follows: anthracycline-based chemotherapy without trastuzumab, trastuzumab with nonanthracycline chemotherapy, anthracyclines followed by trastuzumab (sequential therapy), and chemotherapy without anthracycline/trastuzumab (other chemotherapy). The primary outcome was a composite of hospitalization or emergency room visit for congestive heart failure (CHF), outpatient diagnosis of CHF, or cardiovascular death. A sensitivity analysis limited the outcomes to hospital-based CHF events. Cause-specific hazard models were used accounting for the competing risk of noncardiovascular death. Of 18,540 women included (median age, 54 years; interquartile range, 47 to 63 years), 79% were younger than age 65 years. The cumulative incidence of the primary outcome was 3.08% (95% CI, 2.81% to 3.36%) by 3 years of follow-up, whereas in an age-matched sample of Ontario women (n = 92,700) without breast cancer, it was 0.96% (95% CI, 0.89% to 1.04%). Compared with those receiving other chemotherapy, patients receiving trastuzumab with nonanthracycline chemotherapy and sequential therapy were at a higher risk of cardiotoxicity (hazard ratio, 1.76 [95% CI, 1.19 to 2.60] and 3.96 [95% CI, 3.01 to 5.22], respectively). Hospital-based CHF events were only increased with sequential therapy (hazard ratio, 1.86; 95% CI, 1.07 to 3.22). In women with breast cancer and an age distribution representative of routine clinical practice, trastuzumab-based regimens, including those without anthracyclines, were associated with an increased risk of cardiotoxicity. Sequential therapy increased the risk of hospital-based CHF events.


Effect of dietary fiber content on nutrient digestibility and fecal microbiota composition in growing-finishing pigs.
Microbial population in the gastrointestinal tract plays a central role in health and nutrient digestion. The objective of the present study was to investigate the relationships between microbiota and apparent digestibility coefficients with respect to age and diet. Pigs from Large-White, Duroc or Pietrain breeds were raised under the same housing conditions and fed alternately a low-fiber (LF) and a high-fiber diet (HF) during 4 successive 3-week periods. Data collection for digestibility measurements was achieved during the last week of each period. At the end of each period, fecal microbiota was collected for 16S rRNA gene sequencing. The microbiota remained stable across periods whereas digestibility of energy, crude proteins and cell wall components increased. The microbiota was resilient to diet effect and pigs fed the LF diet were discriminated to those fed the HF diet using 31 predicting OTUs with a mean classification error-rate of 3.9%. Clostridiaceae and Turicibacter were negatively correlated whereas Lactobacillus was positively correlated with protein and energy digestibility coefficients in the LF group. In addition, Lachnospiraceae and Prevotella were negatively correlated with cell wall component digestibility. In contrast, no significant correlation was found between microbiota composition and digestibility coefficients when pigs were fed the HF diet. Interestingly, it was also no longer possible to distinguish animals from different breeds once the animals were fed a HF diet, so that the microbiota could only trace the breed origin in the first period and in the LF group. In our experimental conditions, 3 weeks of adaptation to a new diet seems to be sufficient to observe resilience in growing pigs' microbiota. We demonstrated that fecal microbiota can be used to classify pigs according to their dietary treatment. Some bacteria are favorable or unfavorable to digestibility. This suggests that manipulations of bacterial populations can improve digestibility and feed efficiency.


How should we measure maternal mortality in the developing world? A comparison of household deaths and sibling history approaches.
A reduction in the maternal mortality ratio (MMR) is one of six health-related Millennium Development Goals (MDGs). However, there is no consensus about how to measure MMR in the many countries that do not have complete registration of deaths and accurate ascertainment of cause of death. In this study, we compared estimates of pregnancy-related deaths and maternal mortality in a developing country from three different household survey measurement approaches: a module collecting information on deaths of respondents' sisters; collection of information about recent household deaths with a time-of-death definition of maternal deaths; and a verbal autopsy instrument to identify maternal deaths. We used data from a very large nationally-representative household sample survey conducted in Bangladesh in 2001. A total of 104 323 households were selected for participation, and 99 202 households (95.1% of selected households, 98.8% of contacted households) were successfully interviewed. The sisterhood and household death approaches gave very similar estimates of all-cause and pregnancy-related mortality; verbal autopsy gave an estimate of maternal deaths that was about 15% lower than the pregnancy-related deaths. Even with a very large sample size, however, confidence intervals around mortality estimates were similar for all approaches and exceeded +/- 15%. Our findings suggest that with improved training for survey data collectors, both the sisterhood and household deaths methods are viable approaches for measuring pregnancy-related mortality. However, wide confidence intervals around the estimates indicate that routine sample surveys cannot provide the information needed to monitor progress towards the MDG target. Other approaches, such as inclusion of questions about household deaths in population censuses, should be considered.


Purinergic signaling in the gastrointestinal tract.
Geoffrey Burnstock completed a BSc at King's College London and a PhD at University College London. He held postdoctoral fellowships with Wilhelm Feldberg (National Institute for Medical Research), Edith Bülbring (University of Oxford) and C. Ladd Prosser (University of Illinois). He was appointed to a Senior Lectureship in Melbourne University in 1959 and became Professor and Chairman of Zoology in 1964. In 1975 he became Head of Department of Anatomy and Developmental Biology at UCL and Convenor of the Center of Neuroscience. He has been Director of the Autonomic Neuroscience Institute at the Royal Free Hospital School of Medicine since 1997. He was elected to the Australian Academy of Sciences in 1971, the Royal Society in 1986, the Academy of Medical Sciences in 1998 and an Honorary Fellow of the Royal College of Surgeons and the Royal College of Physicians in 1999 and 2000. He was awarded the Royal Society Gold Medal in 2000. He is editor-in-chief of the journals Autonomic Neuroscience and Purinergic Signalling and on the editorial boards of many other journals. Geoffrey Burnstock's major research interest has been autonomic neurotransmission and he is best known for his seminal discovery of purinergic transmission and receptors, their signaling pathways and functional relevance. He has supervised over 100 PhD and MD students and published over 1400 original papers, re-views and books. He was first in the Institute of Scientific Information list of most cited scientists in Pharmacology and Toxicology from 1994-2004 [59.083 citations (March 2011) and an h-index of 109].


Adalimumab or infliximab as monotherapy, or in combination with an immunomodulator, in the treatment of Crohn's disease.
The comparative efficacy of adalimumab (ADA) and infliximab (IFX) in Crohn's disease, and the benefit of initial combotherapy with an immunomodulator, are debated. To assess the best anti-TNF treatment regimens in Crohn's disease. We included 906 biologic-naïve Crohn's disease patients [median age, 31 years (24-41)] and performed a retrospective analysis of 1284 therapeutic exposures to ADA (n = 521) or IFX (n = 763) between 2006 and 2015. An immunomodulator was associated during the first 4-6 months (initial combotherapy) during 706 therapeutic exposures (55%). Median duration of anti-TNF therapy was 39 months (IQR 17-67). Primary outcomes were 6-month and 2-year response rates and drug survival. Logistic regression with propensity scoring and Cox proportional hazard analysis determined variables associated with outcomes. The response rates at 6 months and 2 years were 64% and 44% on ADA mono, 86% and 70% on ADA combo, 72% and 45% on IFX mono, and 84% and 68% on IFX combotherapy, respectively. Differences between ADA and IFX were not significant, whereas combotherapy was superior to monotherapy (P < 0.001). Drug survival was longer with combotherapy vs. monotherapy [adjusted hazard ratio 2.17 (1.72-2.70)] and not significantly different between ADA and IFX. During subsequent anti-TNF exposures, IFX combotherapy fared better than other groups regarding response rates, drug survival, disease activity, hospitalisations and abdominal surgery. In this retrospective analysis of a large tertiary centre cohort of Crohn's disease patients, ADA and IFX had similar efficacy as first line treatment, while initial combotherapy with an immunomodulator improved all outcome measures.


Carcinosarcoma prostate with osteosarcomatous differentiation: a rare de novo presentation.
Carcinosarcoma is a rare histological event in the history of prostatic malignancies. Historically aggressive tumours with dismal outcomes reported in scarce literature available so far. Very few recent studies suggest good outcomes with modern era surgery and radiotherapy techniques in localised disease. The case presented here had no history of known risk factors like prior adenocarcinoma or prior radiation therapy. This case presented with obstructive urinary symptoms with no prostate-specific antigen elevation, diagnosed with imaging, managed aggressively with robotic surgery. Detailed immunohistochemistry and pathological review suggested diagnosis as carcinosarcoma with osteosarcomatous differentiation. Very rare such cases were reported in the past with complete clinical, radiological, pathological details and managed aggressively with good outcomes. The patient is disease free after 6 months of follow-up.


The mannose cap of mycobacterial lipoarabinomannan does not dominate the Mycobacterium-host interaction.
Pathogenic mycobacteria have the ability to persist in phagocytic cells and to suppress the immune system. The glycolipid lipoarabinomannan (LAM), in particular its mannose cap, has been shown to inhibit phagolysosome fusion and to induce immunosuppressive IL-10 production via interaction with the mannose receptor or DC-SIGN. Hence, the current paradigm is that the mannose cap of LAM is a crucial factor in mycobacterial virulence. However, the above studies were performed with purified LAM, never with live bacteria. Here we evaluate the biological properties of capless mutants of Mycobacterium marinum and M. bovis BCG, made by inactivating homologues of Rv1635c. We show that its gene product is an undecaprenyl phosphomannose-dependent mannosyltransferase. Compared with parent strain, capless M. marinum induced slightly less uptake by and slightly more phagolysosome fusion in infected macrophages but this did not lead to decreased survival of the bacteria in vitro, nor in vivo in zebra fish. Loss of caps in M. bovis BCG resulted in a sometimes decreased binding to human dendritic cells or DC-SIGN-transfected Raji cells, but no differences in IL-10 induction were observed. In mice, capless M. bovis BCG did not survive less well in lung, spleen or liver and induced a similar cytokine profile. Our data contradict the current paradigm and demonstrate that mannose-capped LAM does not dominate the Mycobacterium-host interaction.


Building kit for metal cation binding sites in proteins.
Starting with conformations of calcium-binding sites in parvalbumin and integrin (representative structures of EF-hand and calcium blade zones, respectively) we introduce four new different local Ca2+-recognition units in proteins: a one-residue unit type I (ORI); a three-residue unit type I (TRI); a one-residue unit type II (ORII) and a three-residue unit type II (TRII). Based on the amount and nature of variable atoms, the type I and II units theoretically can have four and twelve variants, respectively. Analysis of known "Ca2+-bound functional niches" in proteins revealed presence of almost all possible variants of Ca2+-recognition units in actual structures. Parvalbumin, integrin alpha-IIb and sixteen other proteins with different Ca2+-bound functional niches contain various consecutively joined combinations of OR(I/II) and TR(I/II) units. Such a OR(I/II)+TR(I/II) joint unit forms a tripeptide, which uses three main-chain atoms for metal binding: nitrogenn (Donor), oxygenn (Acceptor) and nitrogenn+2 (Donor). Thus, taken together, the described ORI, TRI, ORII and TRII units can serve as elementary blocks to construct more complex calcium recognizing substructures in a variety of calcium binding sites of unrelated proteins.


Lipopolysaccharide-induced sickness behavior in lactating rats decreases ultrasonic vocalizations and exacerbates immune system activity in male offspring.
The present study analyzed the effects of lipopolysaccharide (LPS) on maternal behavior during lactation and possible correlations with changes in emotional and immune responses in offspring. Lactating rats received 100 μg/kg LPS, and the control group received saline solution on lactation day (LD) 3. Maternal general activity and maternal behavior were observed on LD5 (i.e. the day that the peak of fever occurred). In male pups, hematological parameters and ultrasonic vocalizations (USVs) were assessed on LD5. At weaning, an additional dose of LPS (50 µg/kg, i.p.) was administered in male pups, and open-field behavior, oxidative burst and phagocytosis were evaluated. A reduction in the time in which dams retrieved the pups was observed, whereas no effects on maternal aggressive behavior were found. On LD5, a reduction of the frequency of USVs was observed in pups, but no signs of inflammation were found. At weaning, an increase in immune system activity was observed, but no differences in open-field behavior were found. These results indicate that inflammation in lactating mothers disrupted mother/pup interactions and may have produced short- and long-term effects on pup behavior as well as biological pathways that modulate inflammatory responses to bacterial endotoxin challenge in pups.


Photophysical and Cellular Imaging Studies of Brightly Luminescent Osmium(II) Pyridyltriazole Complexes.
The series of complexes [Os(bpy)3- n(pytz) n][PF6]2 (bpy = 2,2'-bipyridyl, pytz = 1-benzyl-4-(pyrid-2-yl)-1,2,3-triazole, 1 n = 0, 2 n = 1, 3 n = 2, 4 n = 3) were prepared and characterized and are rare examples of luminescent 1,2,3-triazole-based osmium(II) complexes. For 3 we present an attractive and particularly mild preparative route via an osmium(II) η6-arene precursor circumventing the harsh conditions that are usually required. Because of the high spin-orbit coupling constant associated with the Os(II) center the absorption spectra of the complexes all display absorption bands of appreciable intensity in the range of 500-700 nm corresponding to spin-forbidden ground-state-to-3MLCT transitions (MLCT = metal-to-ligand charge transfer), which occur at significantly lower energies than the corresponding spin-allowed 1MLCT transitions. The homoleptic complex 4 is a bright emitter (λmaxem = 614 nm) with a relatively high quantum yield of emission of ∼40% in deoxygenated acetonitrile solutions at room temperature. Water-soluble chloride salts of 1-4 were also prepared, all of which remain emissive in aerated aqueous solutions at room temperature. The complexes were investigated for their potential as phosphorescent cellular imaging agents, whereby efficient excitation into the 3MLCT absorption bands at the red side of the visible range circumvents autofluorescence from biological specimens, which do not absorb in this region of the spectrum. Confocal microscopy reveals 4 to be readily taken up by cancer cell lines (HeLa and EJ) with apparent lysosomal and endosomal localization, while toxicity assays reveal that the compounds have low dark and light toxicity. These complexes therefore provide an excellent platform for the development of efficient luminescent cellular imaging agents with advantageous photophysical properties that enable excitation and emission in the biologically transparent region of the optical spectrum.


Degradation and leaching of fluroxypyr after application to railway tracks.
Fluroxypyr is an auxin-type herbicide used for postemergent control of broad-leaved weeds in agriculture and in nonagricultural environments such as railways. The overall aim of this study was to assess the potential environmental impact from fluroxypyr application to railway tracks and to elucidate some of the factors that control its environmental fate. In laboratory studies, we examined the degradation of fluroxypyr and the formation of its metabolites fluroxypyr-methoxypyridine (F-MP) and fluroxypyr-pyridinol (F-P) in soil from two Swedish railways. We also investigated the degradation and leaching of fluroxypyr in three different railway plots treated with fluroxypyr (360 g ae ha). The half-life of fluroxypyr in soil samples ranged between 28 and 78 d. An estimated mean 48.6 ± 20% of the fluroxypyr was converted into F-P and 8.0 ± 2% into F-MP. The main metabolite, F-P, was rapidly degraded, with an average half-life of 10 ± 5 d. However, F-MP was not degraded to a significant degree in any sample, resulting in slowly increasing concentrations throughout the experiment. This pattern of relatively rapid degradation of F-P and slow accumulation of F-MP was also observed in the field. The persistent nature of F-MP may be of concern if fluroxypyr is used repeatedly at the same location. Fluroxypyr was detected in the groundwater beneath the track at all three locations studied in concentrations exceeding the EU limit of 0.1 μg L for pesticides in drinking water, and F-P was detected in the groundwater at two of three locations. The most important factor controlling fluroxypyr degradation rate in soil was the soil water content, which modulated microbial activity and presumably also fluroxypyr availability to microorganisms. Our findings imply that fluroxypyr may not be a suitable herbicide for weed control on railway tracks.


Magnitude Estimation with Noisy Integrators Linked by an Adaptive Reference.
Judgments of physical stimuli show characteristic biases; relatively small stimuli are overestimated whereas relatively large stimuli are underestimated (regression effect). Such biases likely result from a strategy that seeks to minimize errors given noisy estimates about stimuli that itself are drawn from a distribution, i.e., the statistics of the environment. While being conceptually well described, it is unclear how such a strategy could be implemented neurally. The present paper aims toward answering this question. A theoretical approach is introduced that describes magnitude estimation as two successive stages of noisy (neural) integration. Both stages are linked by a reference memory that is updated with every new stimulus. The model reproduces the behavioral characteristics of magnitude estimation and makes several experimentally testable predictions. Moreover, the model identifies the regression effect as a means of minimizing estimation errors and explains how this optimality strategy depends on the subject's discrimination abilities and on the stimulus statistics. The latter influence predicts another property of magnitude estimation, the so-called range effect. Beyond being successful in describing decision-making, the present work suggests that noisy integration may also be important in processing magnitudes.


Sequence and structure-activity relationship of a scorpion venom toxin with nitrergic activity in rabbit corpus cavernosum.
An alpha-toxin responsible for nitric oxide (NO) release in rabbit corpus cavernosum (RbCC) was isolated from Tityus serrulatus venom (TSV). The isolated peptide (molecular mass of 7427.66+/-0.15 Da) was identified as Ts3 after determination of Cys residues, N-terminal amino acid analysis, and proteolytic peptide mapping. Ts3 (30 nM) markedly relaxed the RbCC; this response was blocked by the NO synthesis inhibitor N(omega)-nitro-L-arginine methyl ester (100 microM) and the Na+ channel blocker tetrodotoxin (100 nM). Synthetic peptides based on either Ts3 (P1-16, P17-32, P33-48, P49-64, P9-24, P25-40, P41-56, YGLPDKVPTKT) or Bukatoxin (isolated from Buthus martensi Karsch scorpion venom) sequence (Buka11, Buka11-B, PDKVP, PDSEP) were assayed. These peptides slightly relaxed the RbCC, and such an effect was independent of Na+ channel activation or NO release. Our results indicate that Ts3 exerts nitrergic actions and contributes to the relaxing activity of TSV in RbCC, thus providing a valuable tool to investigate the mechanisms underlying nerve activation in erectile tissues, because NO released from nitrergic fibers plays a key role in the erectile process. Our findings revealed the key importance of the Ts3 structure three-dimensional conformation maintenance for biological activity, because linear peptide sequences neither presented substantial relaxations nor was this effect related to nitrergic activity.


Metals, macrocycles and molecular assemblies - macrocyclic complexes in metallo-supramolecular chemistry.
In this tutorial review the use of macrocyclic complexes as building blocks in a selection of supramolecular systems is discussed with emphasis on the properties, such as enhanced stabilities, that cyclic ligands and their complexes may impart on the resulting assemblies. An aim of the review is to exemplify the versatility of macrocyclic ligand complexes for use as components in a range of both discrete and polymeric systems. The use of macrocyclic systems for controlling CuI aggregation, as scaffolds for metal-cluster formation, as the cyclic components in interlocked catenane and rotaxane structures, for constructing assemblies based on macrocycle exo-coordination, for forming columnar stacks, as well as their roles as both structural and redox centres in a range of coordination polymer types are all presented.


A Recurrent CNN for Automatic Detection and Classification of Coronary Artery Plaque and Stenosis in Coronary CT Angiography.
Various types of atherosclerotic plaque and varying grades of stenosis could lead to different management of patients with a coronary artery disease. Therefore, it is crucial to detect and classify the type of coronary artery plaque, as well as to detect and determine the degree of coronary artery stenosis. This paper includes retrospectively collected clinically obtained coronary CT angiography (CCTA) scans of 163 patients. In these, the centerlines of the coronary arteries were extracted and used to reconstruct multi-planar reformatted (MPR) images for the coronary arteries. To define the reference standard, the presence and the type of plaque in the coronary arteries (no plaque, non-calcified, mixed, calcified), as well as the presence and the anatomical significance of coronary stenosis (no stenosis, non-significant, i.e., <50% luminal narrowing, and significant, i.e., ≥50% luminal narrowing) were manually annotated in the MPR images by identifying the start- and end-points of the segment of the artery affected by the plaque. To perform an automatic analysis, a multi-task recurrent convolutional neural network is applied on coronary artery MPR images. First, a 3D convolutional neural network is utilized to extract features along the coronary artery. Subsequently, the extracted features are aggregated by a recurrent neural network that performs two simultaneous multi-class classification tasks. In the first task, the network detects and characterizes the type of the coronary artery plaque. In the second task, the network detects and determines the anatomical significance of the coronary artery stenosis. The network was trained and tested using the CCTA images of 98 and 65 patients, respectively. For detection and characterization of coronary plaque, the method was achieved an accuracy of 0.77. For detection of stenosis and determination of its anatomical significance, the method was achieved an accuracy of 0.80. The results demonstrate that automatic detection and classification of coronary artery plaque and stenosis are feasible. This may enable automated triage of patients to those without coronary plaque and those with coronary plaque and stenosis in need for further cardiovascular workup.


Foamy changes of placental cells in probable beta glucuronidase deficiency associated with hydrops fetalis.
Mucopolysaccharidosis type VII (MPS VII, beta glucuronidase deficiency) has been described in association with non-immune hydrops fetalis. Three consecutive pregnancies in an itinerant family, which resulted in stillbirths caused by non-immune hydrops are described. The parents were closely related and there was a strong family history of storage disorders. The main clue to the diagnosis, however, came from the presence of pronounced foamy cytoplasmic change in the villous Hofbauer cells of the placenta. This raised the possibility of an inherited metabolic storage disorder. The parents were subsequently shown to have beta glucuronidase activities in the heterozygous range in leucocytes and fibroblasts which suggested that the non-immune hydrops was caused by beta glucuronidase deficiency.


Heat/mortality sensitivities in Los Angeles during winter: a unique phenomenon in the United States.
Extreme heat is often associated with elevated levels of human mortality, particularly across the mid-latitudes. Los Angeles, CA exhibits a unique, highly variable winter climate, with brief periods of intense heat caused by downsloping winds commonly known as Santa Ana winds. The goal is to determine if Los Angeles County is susceptible to heat-related mortality during the winter season. This is the first study to specifically evaluate heat-related mortality during the winter for a U.S. city. Utilizing the Spatial Synoptic Classification system in Los Angeles County from 1979 through 2010, we first relate daily human mortality to synoptic air mass type during the winter season (December, January, February) using Welch's t-tests. However, this methodology is only somewhat effective at controlling for important inter- and intra-annual trends in human mortality unrelated to heat such as influenza outbreaks. As a result, we use distributed lag nonlinear modeling (DLNM) to evaluate if the relative risk of human mortality increases during higher temperatures in Los Angeles, as the DLNM is more effective at controlling for variability at multiple temporal scales within the human mortality dataset. Significantly higher human mortality is uncovered in winter when dry tropical air is present in Los Angeles, particularly among those 65 years and older (p < 0.001). The DLNM reveals the relative risk of human mortality increases when above average temperatures are present. Results are especially pronounced for maximum and mean temperatures, along with total mortality and those 65 + . The discovery of heat-related mortality in winter is a unique finding in the United States, and we recommend stakeholders consider warning and intervention techniques to mitigate the role of winter heat on human health in the County.


Low levels of the omega-3 index are associated with sudden cardiac arrest and remain stable in survivors in the subacute phase.
In previous studies, low blood levels of n-3 fatty acids (FA) have been associated with increased risk of cardiac death, and the omega-3 index (red blood cell (RBC) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) expressed as weight percentage of total FA) has recently been proposed as a new risk factor for death from coronary artery disease, especially following sudden cardiac arrest (SCA). As blood samples often haven been harvested after the event, the aim of our study was to evaluate the stability of RBC fatty acids following SCA. The total FA profile, including the omega-3 index, was measured three times during the first 48 h in 25 survivors of out-of-hospital cardiac arrest (OHCA), in 15 patients with a myocardial infarction (MI) without SCA and in 5 healthy subjects. We could not demonstrate significant changes in the FA measurements in any of the groups, this also applied to the omega-6/omega-3 ratio and the arachidonic acid (AA)/EPA ratio. Furthermore, we compared the omega-3 index in 14 OHCA-patients suffering their first MI with that of 185 first-time MI-patients without SCA; mean values being 4.59% and 6.48%, respectively (p = 0.002). In a multivariate logistic regression analysis, a 1% increase of the omega-3 index was associated with a 58% (95% CI: 0.25-0.76%) reduction in risk of ventricular fibrillation (VF). In conclusion, the omega-3 index remained stable after an event of SCA and predicted the risk of VF.


Two-center interference in high-order harmonic generation from heteronuclear diatomic molecules.
Two-center interference for heteronuclear diatomic molecules (HeDM) is investigated. The minimum in the high-order harmonic spectrum, as a consequence of the destructive interference, is shifted to lower harmonic orders compared with that in a homonuclear case. This phenomenon is explained by performing phase analysis. It is found that, for an HeDM, the high harmonic spectrum contains information not only on the internuclear separation but also on the properties of the two separate centers, which implies the potential application of estimating the asymmetry of molecules and judging the linear combination of atomic orbitals (LCAO) for the highest occupied molecular orbital (HOMO). Moreover, the possibility to monitor the evolution of HOMO itself in molecular dynamics is also promised.