Expanding role of beta-blockade in the management of chronic heart failure.
Although recent advances have been made in the treatment of heart failure, this disease continues to result in significant morbidity and mortality. Among the negative effects associated with progression of heart failure are decline in myocardial reserve, decreased exercise tolerance, decreased contractile function, and altered cardiac gene expression. Guidelines recommend neurohormonal antagonists for treatment and stress the importance of angiotensin-converting enzyme inhibition and beta-blockade in reversing the cardiac remodeling process. beta-Blockade slows or reverses the adverse effects resulting from chronic adrenergic stimulation. Traditionally, beta-blockers were reserved for mild-to-moderate heart failure, based on evidence from large, randomized clinical trials showing their positive effects on myocardial function and clinical outcomes. More recently, clinical data reveal that the agents can be expanded to patients with severe heart failure and those with left ventricular systolic dysfunction after myocardial infarction. Individual beta-blocking agents vary in their pharmacology and dosing requirements. These variations may influence treatment decisions and affect clinical measurements of left ventricular function and ventricular remodeling.


Global selection on sucrose synthase haplotypes during a century of wheat breeding.
Spike number per unit area, number of grains per spike, and thousand kernel weight (TKW) are important yield components. In China, increases in wheat (Triticum aestivum) yields are mainly due to increases in grain number per spike and TKW. TKW mainly depends on starch content, as starch accounts for about 70% of the grain endosperm. Sucrose synthase catalysis is the first step in the conversion of sucrose to starch, that is, the conversion of sucrose to fructose and UDP-glucose by the wheat sucrose synthase genes (TaSus1 and TaSus2) that are located on chromosomes 7A/7B/7D and 2A/2B/2D, respectively. A total of 1,520 wheat accessions were genotyped at the six loci. Two, two, five, and two haplotypes were identified at the TaSus2-2A, TaSus2-2B, TaSus1-7A, and TaSus1-7B loci, respectively. Their main variations were detected within the introns. Significant differences between the haplotypes correlated with TKW differences among 348 modern Chinese cultivars from the core collection. Frequency changes for favored haplotypes showed gradual increases in cultivars released since beginning of the last century in China, Europe, and North America. Geographic distributions and time changes of favored haplotypes were characterized in six major wheat production regions worldwide. Strong selection bottlenecks to haplotype variations occurred at polyploidization and domestication and during breeding of wheat. Genetic-effect differences between haplotypes at the same locus influence the selection time and intensity. This work shows that the endosperm starch synthesis pathway is a major target of indirect selection in global wheat breeding for higher yield.


Intrinsic, management, and nutritional factors associated with equine motor neuron disease.
To identify intrinsic, management, nutritional, and environmental risk factors associated with equine motor neuron disease (EMND) and to determine whether epidemiologic evidence supports oxidative stress as a risk factor for developing EMND. Case-control study. 87 horses with EMND and 259 control horses. Information concerning each horse's history of exposure to multiple environmental factors prior to developing EMND was obtained by means of a questionnaire or personal interview. Exposure histories of horses with EMND and control horses were compared, and the association of each risk factor with EMND was evaluated, using logistic regression analysis. Factors significantly associated with risk of developing EMND included age, breed of horse, duration of residence at the farm, not vaccinating against rabies, and certain feeding practices. Horses that were exercised on green pasture or in grass paddocks were less likely to develop EMND, compared with horses that were exercised in dirt pad-docks. Feeding complete pelleted feed as the only source of concentrate or combined with sweet feed was associated with a significant increase in the risk of EMND. Supplementary feeding of vitamin and mineral mixtures not formulated to provide vitamin E or selenium was associated with increased risk of EMND. Horses with a history of cribbing or coprophagia were also at higher risk of developing EMND. Several husbandry practices and intrinsic characteristics of horses appear to modify the risk of EMND. The relationship of specific nutritional factors to EMND supports the hypothesis that a deficiency of vitamin E contributes to the disease.


Efficacy of dexmedetomidine on postoperative shivering: a meta-analysis of clinical trials.
Shivering is a frequent complication in the postoperative period. The aim of the current meta-analysis was to assess the efficacy of dexmedetomidine on postoperative shivering. Two researchers independently searched PubMed, EMBASE and the Cochrane Central Register of Controlled Trials for controlled clinical trials. The meta-analysis was performed by Review Manager. Thirty-nine trials with 2,478 patients were included in this meta-analysis. Dexmedetomidine reduced postoperative shivering compared with placebo (risk ratio [RR] = 0.26; 95% confidence interval [CI]: 0.20 to 0.34), with a minimum effective dose of 0.5 µg·kg(-1) (RR = 0.36; 95% CI: 0.21 to 0.60). The anti-shivering effect can be achieved both intravenously and epidurally when administered within two hours prior to the end of surgery. The efficacy of dexmedetomidine was similar to widely used anti-shivering agents, such as fentanyl, meperidine, tramadol, clonidine and so on; however, dexmedetomidine may increase the incidence of sedation, hypotension, bradycardia and dry mouth. The present meta-analysis indicates that dexmedetomidine shows superiority over placebo, but not over other anti-shivering agents. Therefore, considering its high price and potential adverse events, dexmedetomidine may not be appropriate solely for the purpose of the prevention of postoperative shivering.


Secondary glioblastoma multiforme in a child with disseminated juvenile pilocytic astrocytoma.
Secondary glioblastoma multiforme (sGBM) can occur after a long latency period following radiation treatment of various diseases including brain tumors, leukemia, and more benign disorders like tinea capitis. Outcomes of radiation-induced sGBM remain poor in both children and adults. We report a case of a 16-year-old girl with a history of disseminated juvenile pilocytic astrocytoma treated with chemotherapy and craniospinal radiation 9 years prior who developed sGBM in the absence of a tumor predisposition syndrome. She presented with a several-week history of headaches and no acute findings on computed tomography compared to baseline neuroimaging 3 months prior. Repeat computed tomography performed just 3 weeks later for worsening headaches revealed a new large posterior fossa tumor where pathology confirmed the diagnosis of sGBM. In spite of maximal surgical resection, reirradiation, and adjuvant chemotherapy, she died 1 year postdiagnosis. Our case highlights the potential late effects of high-dose cranial radiation, how symptomatology may precede neuroimaging findings, and the rapid formation of sGBM that mirrors that of de novo Glioblastoma Multiforme.


Angiography with fluorescein-labeled dextrans in a primate model of uveitis.
Sequential fluorescein angiography, using fluorescein-labeled dextran molecules of several different sizes, was carried out in monkeys with intraocular inflammation induced with interphotoreceptor retinoid-binding protein. The vascular leakage seen with the dextrans was compared with that seen with standard fluorescein sodium angiography. The angiograms demonstrated that different-sized leaks appear in the retinal vessels in adjacent areas during the course of the inflammation. Most retinal vessels leaked only fluorescein sodium and no dextran of any size, suggesting that it is the unbound fluorescein that leaks out of these vessels and not fluorescein bound to plasma albumin. It was not possible to tell by clinical examination which areas would leak the larger-molecular weight tracers. Ultrastructural studies of the veins leaking the dextrans revealed areas of abnormal endothelial tight junctions, whereas the tight junctions were normal in areas where leakage occurred with fluorescein alone.


Sequential hand hygiene promotion contributes to a reduced nosocomial bloodstream infection rate among very low-birth weight infants: an interrupted time series over a 10-year period.
Sustained high compliance with hand hygiene (HH) is needed to reduce nosocomial bloodstream infections (NBSIs). However, over time, a wash out effect often occurs. We studied the long-term effect of sequential HH-promoting interventions. An observational study with an interrupted time series analysis of the occurrence of NBSI was performed in very low-birth weight (VLBW) infants. Interventions consisted of an education program, gain-framed screen saver messages, and an infection prevention week with an introduction on consistent glove use. A total of 1,964 VLBW infants admitted between January 1, 2002, and December 31, 2011, were studied. The proportion of infants with ≥1 NBSI decreased from 47.6%-21.2% (P < .01); the number of NBSIs per 1,000 patient days decreased from 16.8-8.9 (P < .01). Preintervention, the number of NBSIs per 1,000 patient days significantly increased by 0.74 per quartile (95% confidence interval [CI], 0.27-1.22). The first intervention was followed by a significantly declining trend in NBSIs of -1.27 per quartile (95% CI, -2.04 to -0.49). The next interventions were followed by a neutral trend change. The relative contributions of coagulase-negative staphylococci and Staphylococcus aureus as causative pathogens decreased significantly over time. Sequential HH promotion seems to contribute to a sustained low NBSI rate.


Magnetic induction tomography: comparison of the image quality using different types of receivers.
Magnetic induction tomography is used to image the electrical properties inside a region of interest. The systems differ in the construction of the receiver channels which can be composed of coils or gradiometers. We will compare and discuss the image quality subject to two different types of receivers, different arrangements for the exciters and receivers and different signal-to-noise ratios. In order to evaluate the image quality, the point-spread function (PSF) was determined which is used for the calculation of the resolution and the correctness of the location of a perturbation. The results show that the PSF depends on (a) the location inside the object, (b) the type of receivers and (c) the configuration used, especially the location of the receiving and excitation channels. According to this, the local resolution is changed and has the maximum near the border of the object and decreases towards the centre of the object. In addition, the evaluation of the PSF shows a dislocation with respect to the underlying point-source position.


Cascades, thermalization, and eddy viscosity in helical Galerkin truncated Euler flows.
The dynamics of the truncated Euler equations with helical initial conditions are studied. Transient energy and helicity cascades leading to Kraichnan helical absolute equilibrium at small scales, including a linear scaling of the relative helicity spectrum are obtained. Strong helicity effects are found using initial data concentrated at high wave numbers. Using low-wave-number initial conditions, the results of Cichowlas et.al. [Phys. Rev. Lett. 95, 264502 (2005)] are extended to helical flows. Similarities between the turbulent transient evolution of the ideal (time-reversible) system and viscous helical flows are found. Using an argument in the manner of Frisch et. al. [Phys. Rev. Lett. 101, 144501 (2008)], the excess of relative helicity found at small scales in the viscous run is related to the thermalization of the ideal flow. The observed differences in the behavior of truncated Euler and (constant viscosity) Navier-Stokes are qualitatively understood using the concept of eddy viscosity. The large scales of truncated Euler equations are then shown to follow quantitatively an effective Navier-Stokes dynamics based on a variable (scale dependent) eddy viscosity.


Cooperation of the DnaK and GroE chaperone systems in the folding pathway of plant ferredoxin-NADP+ reductase expressed in Escherichia coli.
The DnaK system is required for the productive folding of pea chloroplast ferredoxin-NADP+ reductase (FNR) expressed in Escherichia coli. The formation of a mature active enzyme was severely impaired in E. coli dnaK, dnaJ or grpE mutants expressing either the cytosolic precursor of the reductase (preFNR) or the mature apoenzyme, and these forms aggregated extensively in these cells. Coexpression of dnaK from a multicopy plasmid in the dnaK-null mutants restored preFNR processing and folding of FNR, rendering a mature-sized active enzyme. Overexpression of GroESL chaperonins failed to prevent preFNR aggregation, but it restored productive folding of FNR in dnaK-null mutants expressing the mature enzyme. Expression of preFNR in OmpT-protease-deficient E. coli cells resulted in the accumulation of the unprocessed precursor in the soluble fraction of the cells. The interaction of this soluble preFNR, but not the mature reductase, with DnaK and GroEL was evidenced by immunoprecipitation studies. We conclude that, in addition to the GroE chaperonins [Carrillo, N., Ceccarelli, E. A., Krapp, A. R., Boggio, S., Ferreyra, R. G. & Viale, A. M. (1992) J. Biol. Chem. 267, 15537-15541], the DnaK chaperone system plays a crucial role in the folding pathway of FNR.


[Use of Pseudomonas bacteria strains for determining water-assimilable carbon].
The ability of 12 Pseudomonas putida and Pseudomonas fluorescens strains for growth in the tap water has been studied. All of them grew well in pasteurized Kyiv tap water at 500 colony-forming units--CFU/ml inoculum reaching the maximal titre 1.7 x 10(4)-4.6 x 10(6) CFU/ml after 240 hours of incubation. The most intensive growth was observed in strain P. putida UCM B-143, capable to assimilation of 84 different sources of carbon nutrition. The linear dependence between strain 143 growth and acetate carbon concentration in the interval of 10 to 60 microg/l has been demonstrated in deionized water with addition of necessary mineral salts; its yield was 4.5 x 10(6) CFU/microg of assimilable acetate carbon (AOC). Determination of AOC of water using standard methods, standard strain P. fluorescens UCM B-385 (P17, ATCC 49642) and selected strain P. putida B-143 gave the similar values of AOC quantities in the Kyiv tap water (536 and 600 microg of AOC in 11, accordingly). Viability of strain 143 grown in the artesian water decreased more quickly than that of the standard strain which is possibly connected with origin of both cultures: strain 143 was isolated from wheat rhizosphere while strain P17--from the drinking water.


Role of the oligopeptide permease ABC Transporter of Moraxella catarrhalis in nutrient acquisition and persistence in the respiratory tract.
Moraxella catarrhalis is a strict human pathogen that causes otitis media in children and exacerbations of chronic obstructive pulmonary disease in adults, resulting in significant worldwide morbidity and mortality. M. catarrhalis has a growth requirement for arginine; thus, acquiring arginine is important for fitness and survival. M. catarrhalis has a putative oligopeptide permease ABC transport operon (opp) consisting of five genes (oppB, oppC, oppD, oppF, and oppA), encoding two permeases, two ATPases, and a substrate binding protein. Thermal shift assays showed that the purified recombinant substrate binding protein OppA binds to peptides 3 to 16 amino acid residues in length regardless of the amino acid composition. A mutant in which the oppBCDFA gene cluster is knocked out showed impaired growth in minimal medium where the only source of arginine came from a peptide 5 to 10 amino acid residues in length. Whether methylated arginine supports growth of M. catarrhalis is important in understanding fitness in the respiratory tract because methylated arginine is abundant in host tissues. No growth of wild-type M. catarrhalis was observed in minimal medium in which arginine was present only in methylated form, indicating that the bacterium requires l-arginine. An oppA knockout mutant showed marked impairment in its capacity to persist in the respiratory tract compared to the wild type in a mouse pulmonary clearance model. We conclude that the Opp system mediates both uptake of peptides and fitness in the respiratory tract.


Ileal-lymphoid-nodular hyperplasia, non-specific colitis, and pervasive developmental disorder in children.
We investigated a consecutive series of children with chronic enterocolitis and regressive developmental disorder. 12 children (mean age 6 years [range 3-10], 11 boys) were referred to a paediatric gastroenterology unit with a history of normal development followed by loss of acquired skills, including language, together with diarrhoea and abdominal pain. Children underwent gastroenterological, neurological, and developmental assessment and review of developmental records. Ileocolonoscopy and biopsy sampling, magnetic-resonance imaging (MRI), electroencephalography (EEG), and lumbar puncture were done under sedation. Barium follow-through radiography was done where possible. Biochemical, haematological, and immunological profiles were examined. Onset of behavioural symptoms was associated, by the parents, with measles, mumps, and rubella vaccination in eight of the 12 children, with measles infection in one child, and otitis media in another. All 12 children had intestinal abnormalities, ranging from lymphoid nodular hyperplasia to aphthoid ulceration. Histology showed patchy chronic inflammation in the colon in 11 children and reactive ileal lymphoid hyperplasia in seven, but no granulomas. Behavioural disorders included autism (nine), disintegrative psychosis (one), and possible postviral or vaccinal encephalitis (two). There were no focal neurological abnormalities and MRI and EEG tests were normal. Abnormal laboratory results were significantly raised urinary methylmalonic acid compared with age-matched controls (p=0.003), low haemoglobin in four children, and a low serum IgA in four children. We identified associated gastrointestinal disease and developmental regression in a group of previously normal children, which was generally associated in time with possible environmental triggers.


Proteomics in pediatric cystic craniopharyngioma.
Adamantinomatous craniopharyngioma (ACP) is still often burdened by a poor prognosis in children as far as the risk of recurrence and the quality of life are concerned. Therefore, many efforts are now dedicated to investigate the molecular characteristics of this tumor aiming at finding new therapeutic options. ACP is prevalently a cystic lesion so that an increasing number of researches are focused on the analysis of its cystic content. In the present article, the main results of the current proteomic analysis (PA) on the ACP fluid are summarized. Both "bottom-up" and "top-down" approaches have been utilized. In the bottom-up approach, proteins and peptides are enzymatically or chemically digested prior to liquid chromatography and mass spectrometry analyses. The bottom-up approach pointed out several proteins of the inflammation (namely, α2-HS-glycoprotein, α1-antichymotrypsin and apolipoproteins) as possibly involved in the genesis and growth of the cystic component of ACP. The top-down strategy analyzes proteins and peptides in the intact state, making it particularly suitable for the identification of peptides and low molecular weight proteins and for the characterization of their possible isoforms and post-translational modifications. The top-down approach disclosed the presence of the thymosin β family. Thymosin β4, in particular, which is involved in the cytoskeleton organization and migration of several tumors, could play a role in the progression of ACP. Finally, PA was utilized to investigate alterations in cyst fluid character after treatment with interferon-α. The analyzed samples showed a progressive reduction of the levels of α-defensins (proteins involved in the inflammatory-mediated response) after the intracystic injection of interferon-α, thus reinforcing the hypothesis that inflammation contributes to ACP cyst pathogenesis. Additional studies on the solid component of ACP are still necessary to further validate the previous results and to identify possible markers for targeted therapy.


Inhibition of human immunodeficiency virus type I reverse transcriptase by suramin-related compounds.
Ninety analogues of suramin have been examined for their ability to inhibit the exogenous reverse transcriptase (RT) of human immunodeficiency virus type I (HIV-I). Of these compounds, 57 inhibited the poly(rC).oligo(dG)-dependent RT activity. Three classes of dose-response curves could be discriminated. Allocation of a compound to one class did not correspond with obvious structural features. Twenty-four substances were superior to suramin in our RT inhibition assay. The RT-inhibitory activity of these compounds did not correlate with their effect against filariae or trypanosomes. Preliminary antiviral evaluation in susceptible human T cells inoculated with HIV-I demonstrated in vitro therapeutic efficacy for some compounds with lower drug-related cellular toxicity than suramin. Certain structural features relevant for the RT-inhibitory effect of these compounds were recognized. Predictions are made for the design of more effective RT inhibitors. Such compounds will help to understand the molecular mechanism of reverse transcription and might be useful in the therapy of retroviral infections.


Type I interferons (alpha/beta) in immunity and autoimmunity.
The significance of type I interferons (IFN-alpha/beta) in biology and medicine renders research on their activities continuously relevant to our understanding of normal and abnormal (auto) immune responses. This relevance is bolstered by discoveries that unambiguously establish IFN-alpha/beta, among the multitude of cytokines, as dominant in defining qualitative and quantitative characteristics of innate and adaptive immune processes. Recent advances elucidating the biology of these key cytokines include better definition of their complex signaling pathways, determination of their importance in modifying the effects of other cytokines, the role of Toll-like receptors in their induction, their major cellular producers, and their broad and diverse impact on both cellular and humoral immune responses. Consequently, the role of IFN-alpha/beta in the pathogenesis of autoimmunity remains at the forefront of scientific inquiry and has begun to illuminate the mechanisms by which these molecules promote or inhibit systemic and organ-specific autoimmune diseases.


Further characterization of a type III secretion system (T3SS) and of a new effector protein from a clinical isolate of Aeromonas hydrophila--part I.
A type III secretion system (T3SS)-associated cytotoxin, AexT, with ADP-ribosyltransferase activity and homology to Pseudomonas aeruginosa bifuncational toxins ExoT/S, was recently identified from a fish pathogen Aeromonas salmonicida. In this study, we reported the molecular characterization of an aexT-like toxin gene (designated as aexU) from a diarrheal isolate SSU of A. hydrophila. The aexU gene was 1539bp in length and encoded a protein of 512 amino acid (aa) residues. The NH(2)-terminus of AexU (aa residues 1-231) exhibited a 67% homology with the NH(2)-terminus of AexT from A. salmonicida. Importantly, its COOH-terminus (aa residues 232-512) had no homology with any known functional proteins in the database; however, the full-length AexU retained ADP-ribosyltransferase activity. The expression and subsequent secretion of AexU was T3SS dependent, as inactivation of the ascV gene that codes for an inner-membrane component of the T3SS channel from the wild-type (WT) bacterium, blocked translocation of AexU in HT-29 human colonic epithelial cells. We provided evidence that inactivation of acrV and axsE genes (homologs of lcrV and exsE in Yersinia species and P. aeruginosa, respectively) from A. hydrophila SSU, altered expression and/or secretion of AexU. We deleted an aexU gene from the WT, as well as from the DeltaaopB mutant, of A. hydrophila, generating a single knockout (DeltaaexU) and a double knockout mutant, DeltaaopB/DeltaaexU. Increased phagocytosis was observed in RAW264.7 murine macrophages infected with the DeltaaopB/DeltaaexU mutant, as compared to macrophages when infected with the parental DeltaaopB strain. Further, mice infected with the DeltaaexU mutant had a 60% survival rate, compared to animals infected with the WT or the DeltaaexU-complemented strain that caused 90-100% of the animals to die at a 2-3 LD(50s) dose. Immunization of mice with the recombinant AexU protected them from subsequent lethal challenge dose by the WT bacterium. Finally, we detected specific anti-AexU antibodies in the sera of mice that survived challenge by the WT bacterium, which may indicate that AexU plays an important role in the pathogenesis of Aeromonas infections.


Tissue expansion in the reconstruction of Tessier craniofacial clefts: a series of 17 patients.
Tessier craniofacial clefts are among the most surgically challenging examples of craniofacial dysmorphology. These clefts are characterized by hypoplasia of soft-tissue and skeletal elements throughout the three-dimensional extent of the cleft. Whereas bone grafting and craniofacial osteotomies have been successful toward correcting the underlying skeletal abnormalities, the ultimate success of these reconstructions has been limited by the deficiency of skin and soft tissue. This deficiency demands reconstruction ideally with tissue of like texture, consistency, and, especially in the face, color. Craniofacial tissue expansion was used toward reconstructing these facial clefts with like-quality tissue, allowing for tension-free reconstruction after osteotomy and bone grafting. Seventeen patients with Tessier craniofacial clefts underwent preoperative craniofacial soft-tissue expansion in the surgical management of their clefts. Tissue expansion was used in the primary correction of facial clefts in eight patients, with nine patients undergoing expansion before secondary surgery. In this series, tissue expansion has evolved as an important element in overcoming the skin and soft-tissue deficiency associated with these clefts, allowing for tension-free closure and improved aesthetic results in these surgically challenging patients.


Optimizing Preseason Training Loads in Australian Football.
To investigate whether preseason training plans for Australian football can be computer generated using current training-load guidelines to optimize injury-risk reduction and performance improvement. A constrained optimization problem was defined for daily total and sprint distance, using the preseason schedule of an elite Australian football team as a template. Maximizing total training volume and maximizing Banister-model-projected performance were both considered optimization objectives. Cumulative workload and acute:chronic workload-ratio constraints were placed on training programs to reflect current guidelines on relative and absolute training loads for injury-risk reduction. Optimization software was then used to generate preseason training plans. The optimization framework was able to generate training plans that satisfied relative and absolute workload constraints. Increasing the off-season chronic training loads enabled the optimization algorithm to prescribe higher amounts of "safe" training and attain higher projected performance levels. Simulations showed that using a Banister-model objective led to plans that included a taper in training load prior to competition to minimize fatigue and maximize projected performance. In contrast, when the objective was to maximize total training volume, more frequent training was prescribed to accumulate as much load as possible. Feasible training plans that maximize projected performance and satisfy injury-risk constraints can be automatically generated by an optimization problem for Australian football. The optimization methods allow for individualized training-plan design and the ability to adapt to changing training objectives and different training-load metrics.


Technical Note: Evaluation of kV CBCT enhancement using a liver-specific contrast agent for stereotactic body radiation therapy image guidance.
To evaluate possible use for cone-beam computed tomography (CBCT) guidance, this phantom study evaluated the contrast enhancement provided by Gadoxetate Disodium (Primovist® CAN/EU, or Eovist® USA, Bayer Healthcare, Leverkusen, Germany), a contrast agent that is taken up selectively by liver cells and is retained for up to an hour. Image quality from CBCT was benchmarked against helical fan-beam computed tomography for two phantom geometries. Concentrations were diluted to 0.0125-0.1 mmol per kilogram of body weight (mmol/kg) corresponding to expected physiological concentrations in the liver. Kilovoltage CBCT imaging parameters of x-ray tube potential, current, and filtration were investigated using clinically available options on a TrueBeam STx linear accelerator CBCT platform. Two phantoms were created, a cylindrical idealized imaging geometry and an ellipsoidal more realistic abdominal geometry. All parameters were optimized according to the contrast-to-noise ratio (CNR) image quality metric, as a function of concentration, following the Rose criterion for CNR. Acceptable CNR was defined as greater than or equal to three, in accordance with the Rose criterion for CNR. These were found in a range of expected liver concentrations of 0.025-0.1 mmol/kg for a tube potential of 100 kVp, half-fan bowtie filtration and tube currents giving exposures between 2025 and 5085 mAs. Linear correlations were found for all CNR as a function of concentration, in agreement with the literature. Based on this phantom study, with appropriate selection of imaging protocol, Gadoxetate Disodium may provide useful liver CBCT enhancement at physiologically achievable liver concentrations.


Overt Infection with Chronic Bee Paralysis Virus (CBPV) in Two Honey Bee Colonies.
Chronic Bee Paralysis Virus (CBPV), a widespread honey bee RNA virus, causes massive worker bee losses, mostly in strong colonies. Two different syndromes, with paralysis, ataxia and flight incapacity on one hand and black hairless individuals with shortened abdomens on the other, can affect a colony simultaneously. This case report presents two Apis mellifera carnica colonies with symptoms of paralysis and hairless black syndrome in 2019. Via RT-PCR, a highly positive result for CBPV was detected in both samples. Further problems, such as a Nosema infection and Varroa infestation, were present in these colonies. Therapy methods were applied to colony 1 comprising queen replacement, shook swarm method and Varroa control, whereas colony 2 was asphyxiated after queen loss and colony weakening. After therapy, colony 1 was wintered without symptoms. Beekeeping and sanitary measures can save a CBPV-infected colony, while further complications result in total colony loss.


Recycling, channeling and heterogeneous protein turnover estimation using a model of whole-body protein turnover based on leucine kinetics in rodents.
In the companion paper, a whole-body, mechanistic model of protein turnover in a rodent was described and evaluated with independent data sets that used the flooding dose method. On the basis of fitted fluxes, the model was able to predict specific radioactivity changes in the protein and free leucine pools and whole-body protein fractional synthesis rate (FSR). In this paper, results of model simulations of specific radioactivity changes in the flooding dose, pulse dose and continuous infusion methods were compared and the influence of recycling, channeling and multiple protein pools on model behavior were analyzed. For all methods, the percentage of channeling must be estimated to determine whether the extracellular or intracellular pool specific radioactivities better approximate the aminoacyl tRNA pool specific radioactivity. Recycling also affects the specific radioactivity of the aminoacyl-tRNA pool and therefore must be estimated. An analysis of fits of the flooding dose data indicated that 100% channeling was occurring, but the percentage of recycling could not be determined. Multiple protein pools turning over at different rates overestimated FSR by 2-3% at early time points (5 min) and underestimated FSR by 3-6% at 60 min in the flooding dose method. For the pulse dose method, FSR was underestimated by 40-50% at 5 min and underestimated by 9-10% at 60 min. An increase in time to measure FSR caused a decrease in the estimate of FSR (18% over 3 h) for the flooding dose method and an increase in the estimate of FSR (144% over 3 h) for the pulse dose method.


Structure of an IkappaBalpha/NF-kappaB complex.
The inhibitory protein, IkappaBalpha, sequesters the transcription factor, NF-kappaB, as an inactive complex in the cytoplasm. The structure of the IkappaBalpha ankyrin repeat domain, bound to a partially truncated NF-kappaB heterodimer (p50/ p65), has been determined by X-ray crystallography at 2.7 A resolution. It shows a stack of six IkappaBalpha ankyrin repeats facing the C-terminal domains of the NF-kappaB Rel homology regions. Contacts occur in discontinuous patches, suggesting a combinatorial quality for ankyrin repeat specificity. The first two repeats cover an alpha helically ordered segment containing the p65 nuclear localization signal. The position of the sixth ankyrin repeat shows that full-length IkappaBalpha will occlude the NF-kappaB DNA-binding cleft. The orientation of IkappaBalpha in the complex places its N- and C-terminal regions in appropriate locations for their known regulatory functions.


Hyperviscosity-related retinopathy and serous macular detachment in Waldenström's macroglobulinemia: A mortal case in 5 years.
To present a 5-year mortal case of Waldenström's macroglobulinemia-related retinopathy and serous macular detachment. A 63-year-old man, with unremarkable medical history, presented with bilateral decreased vision for 2 months. Fundus examination revealed bilateral scattered retinal hemorrhages, exudates, venous tortuosity, and serous macular detachment. Hematologic and biochemistry profiles showed pancytopenia with blood smear demonstrating erythrocyte rouleaux formation. Hyperviscosity syndrome was suspected and later Waldenström's macroglobulinemia was diagnosed by bone marrow biopsy and high concentration of serum IgM. Plasmapheresis and subsequent chemotherapy were arranged. In spite of resolution of most retinal abnormalities, his visual acuity still showed no improvement with a persistent bilateral macular detachment. The patient then died 5 years after the diagnosis of Waldenström's macroglobulinemia. Long-term toxicity of IgM to the retinal pigment epithelium may impede the resolution of the persistent serous macular detachment, resulting in an inability of recovery in his vision. Therefore, early diagnosis and timely reduction of serum paraproteins by plasmapheresis and chemotherapy is critical for preventing permanent damages to patients' health and vision.


Interfacial Rheology of Sterically Stabilized Colloids at Liquid Interfaces and Its Effect on the Stability of Pickering Emulsions.
Particle-laden interfaces can be used to stabilize a variety of high-interface systems, from foams over emulsions to polymer blends. The relation between the particle interactions, the structure and rheology of the interface, and the stability of the system remains unclear. In the present work, we experimentally investigate how micron-sized, near-hard-sphere-like particles affect the mechanical properties of liquid interfaces. In particular, by comparing dried and undried samples, we investigate the effect of aggregation state on the properties of the particle-laden liquid interface and its relation to the stability of the corresponding Pickering emulsions. Partially aggregated suspensions give rise to a soft-solid-like response under shear, whereas for stable PMMA particulate layers a liquid-like behavior is observed. For interfacial creep-recovery measurements, we present an empirical method to correct for the combined effect of the subphase drag and the compliance of the double-wall ring geometry, which makes a significant contribution to the apparent elasticity of weak interfaces. We further demonstrate that both undried and dried PMMA particles can stabilize emulsions for months, dispelling the notion that particle aggregation, in bulk or at the interface, is required to create stable Pickering emulsions. Our results indicate that shear rheology is a sensitive probe of colloidal interactions but is not necessarily a predictor of the stability of interfaces, e.g., in quiescent Pickering emulsions, as in the latter the response to dilatational deformations can be of prime importance.


ATR and ATM regulate the timing of DNA replication origin firing.
Timing of DNA replication initiation is dependent on S-phase-promoting kinase (SPK) activity at discrete origins and the simultaneous function of many replicons. DNA damage prevents origin firing through the ATM- and ATR-dependent inhibition of Cdk2 and Cdc7 SPKs. Here, we establish that modulation of ATM- and ATR-signalling pathways controls origin firing in the absence of DNA damage. Inhibition of ATM and ATR with caffeine or specific neutralizing antibodies, or upregulation of Cdk2 or Cdc7, promoted rapid and synchronous origin firing; conversely, inhibition of Cdc25A slowed DNA replication. Cdk2 was in equilibrium between active and inactive states, and the concentration of replication protein A (RPA)-bound single-stranded DNA (ssDNA) correlated with Chk1 activation and inhibition of origin firing. Furthermore, ATM was transiently activated during ongoing replication. We propose that ATR and ATM regulate SPK activity through a feedback mechanism originating at active replicons. Our observations establish that ATM- and ATR-signalling pathways operate during an unperturbed cell cycle to regulate initiation and progression of DNA synthesis, and are therefore poised to halt replication in the presence of DNA damage.


Glucose, glycation, and RAGE: implications for amplification of cellular dysfunction in diabetic nephropathy.
Receptor for advanced glycation endproducts (RAGE) is a multi-ligand member of the immunoglobulin superfamily of cell surface molecules. Driven by rapid accumulation and expression of key ligands such as advanced glycation endproducts (AGE) and S100/calgranulins in diabetic tissues, upregulation and activation of RAGE magnifies cellular perturbation in tissues affected by hyperglycemia, such as the large blood vessels and the kidney. In the diabetic glomerulus, RAGE is expressed principally by glomerular visceral epithelial cells (podocytes). Blockade of RAGE in the hyperglycemic db/db mouse suppresses functional and structural alterations in the kidney, in the absence of alterations in blood glucose. Recent studies in homozygous RAGE null mice support a key role for RAGE in glomerular perturbation in diabetes. Importantly, beyond diabetes, studies in other settings of glomerulopathies support a critical RAGE-dependent pathway in podocytes linked to albuminuria, mesangial expansion, and glomerular sclerosis. A new paradigm is proposed in glomerular injury, and it is suggested that blockade of the RAGE axis may provide a novel means to prevent irreparable glomerular injury in diabetes and other sclerosing glomerulopathies.


Myotendinous Thermoregulation in National Level Sprinters after a Unilateral Fatigue Acute Bout-A Descriptive Study.
In the last decade there has been a growing interest in infrared thermography in the field of sports medicine in order to elucidate the mechanisms of thermoregulation. The aim of this study was to describe bilateral variations in skin temperature of the anterior thigh and patellar tendon in healthy athletes and to provide a model of baseline tendon and muscle thermoregulation in healthy sprinters following a unilateral isokinetic fatigue protocol. Fifteen healthy national-level sprinters (eleven men and four women), with at least 3 years of athletic training experience of 10-12 h/week and competing in national-level competitions, underwent unilateral isokinetic force testing and electrostimulation in which their body temperature was measured before, during, and after the protocol using an infrared thermographic camera. ANOVA detected a significant difference in the time × side interaction for patellar temperature changes (p ≤ 0.001) and a significant difference in the time/side interaction for quadriceps temperature changes (p ≤ 0.001). The thermal challenge produces homogeneous changes evident in quadriceps areas, but not homogeneous in tendon areas. These data show that metabolic and blood flow changes may depend on the physical and mechanical properties of each tissue. Future research could be conducted to evaluate the predictive value of neuromuscular fatigue in the patellar tendon and quadriceps after exercise in order to optimize post-exercise recovery strategies.


Three years follow-up of pamidronate therapy in two brothers with osteoporosis-pseudoglioma syndrome (OPPG) carrying an LRP5 mutation.
Osteoporosis-pseudoglioma (OPPG) is a rare syndrome characterized by severe osteoporosis and ocular defects caused by homozygotic inactivation mutations in the LRP5 gene. Bisphosphonate has been demonstrated to improve bone mineral density (BMD) in children with OPPG. We present here a 3 years follow-up of two brothers with OPPG carrying a novel mutation in the LRP5 gene, who were treated with intravenous pamidronate. We looked for a mutation in the LRP5 gene in two brothers (12 and 4 years old) with clinical features of OPPG (blindness, low BMD and fragility fractures) and in their consanguineous parents to confirm the diagnosis of OPPG. The patients were treated with bisphosphonate for 3 years. They received 1 mg/kg/day of pamidronate for 2 consecutive days, every 3 months during the first year, and every 4 months in subsequent years. Calcium, phosphorus, total alkaline phosphatase, parathyroid hormone, hepatic transaminases, creatinine and hemogram tests were performed before each infusion. Bone densitometry was performed at baseline and at the end of the follow-up. The affected brothers carry a missense mutation in the third codon of exon 8 (AAT-->ATT) that led to the exchange of an asparagine for an isoleucine (N531I). Both parents were found to be heterozygous for this mutation. The intravenous pamidronate therapy was safe for up to 3 years of use. Moreover, increased BMD and decreased fracture rate were observed in our patients with OPPG.


The Establishment Risk of Lycorma delicatula (Hemiptera: Fulgoridae) in the United States and Globally.
Native to Asia, the spotted lanternfly, Lycorma delicatula (White), is an emerging pest of many commercially important plants in Korea, Japan, and the United States. Determining its potential distribution is important for proactive measures to protect commercially important commodities. The objective of this study was to assess the establishment risk of L. delicatula globally and in the United States using the ecological niche model MAXENT, with a focus on Washington State (WA), where large fruit industries exist. The MAXENT model predicted highly suitable areas for L. delicatula in Asia, Oceania, South America, North America, Africa, and Europe, but also predicted that tropical habitats are not suitable for its establishment, contrary to published information. Within the United States, the MAXENT model predicted that L. delicatula can establish in most of New England and the mid-Atlantic states, the central United States and the Pacific Coast states, including WA. If introduced, L. delicatula is likely to establish in fruit-growing regions of the Pacific Northwest. The most important environmental variables for predicting the potential distribution of L. delicatula were mean temperature of driest quarter, elevation, degree-days with a lower developmental threshold value of 11°C, isothermality, and precipitation of coldest quarter. Results of this study can be used by regulatory agencies to guide L. delicatula surveys and prioritize management interventions for this pest.


[Correction of physical performance of mice with the derivatives of 3-oxypyridine and pyridine].
Effects of 21 phenyl ethyl-substituted derivatives of 3-oxypyridine (3-OP), 6 adamantine derivatives of pyridine (PD) and actoprotectors bemithyl and bromanthane on physical performance (PP) were evaluated during treadmill testing of mice in ordinary conditions and following vibration. It was stated that in ordinary conditions PP was improved by eleven 3-OP derivatives and 3 adamantine PDs; nine 3-OP derivatives and 3 adamantine PDs had a positive effect following vibration. The actoprotective effect of the compounds depends on their chemical composition, dose and a background. A phenyl ethyl-substituted 3-OP derivative (code CK-132) administered in ordinary conditions and following vibration surpassed significantly the other compounds tested or compared with (bemithyl and bromanthane) in this experiment with regard to the spectrum of effective doses (5-100 mg/kg) and the actoprotective activity (PP improved by 32 to 110%). The compound (code CK-132) can be further test validated as an actoprotector against the impacts of vibration.


Properties of LINE-1 proteins and repeat element expression in the context of amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease involving loss of motor neurons and having no known cure and uncertain etiology. Several studies have drawn connections between altered retrotransposon expression and ALS. Certain features of the LINE-1 (L1) retrotransposon-encoded ORF1 protein (ORF1p) are analogous to those of neurodegeneration-associated RNA-binding proteins, including formation of cytoplasmic aggregates. In this study we explore these features and consider possible links between L1 expression and ALS. We first considered factors that modulate aggregation and subcellular distribution of LINE-1 ORF1p, including nuclear localization. Changes to some ORF1p amino acid residues alter both retrotransposition efficiency and protein aggregation dynamics, and we found that one such polymorphism is present in endogenous L1s abundant in the human genome. We failed, however, to identify CRM1-mediated nuclear export signals in ORF1p nor strict involvement of cell cycle in endogenous ORF1p nuclear localization in human 2102Ep germline teratocarcinoma cells. Some proteins linked with ALS bind and colocalize with L1 ORF1p ribonucleoprotein particles in cytoplasmic RNA granules. Increased expression of several ALS-associated proteins, including TAR DNA Binding Protein (TDP-43), strongly limits cell culture retrotransposition, while some disease-related mutations modify these effects. Using quantitative reverse transcription PCR (RT-qPCR) of ALS tissues and reanalysis of publicly available RNA-Seq datasets, we asked if changes in expression of retrotransposons are associated with ALS. We found minimal altered expression in sporadic ALS tissues but confirmed a previous report of differential expression of many repeat subfamilies in C9orf72 gene-mutated ALS patients. Here we extended understanding of the subcellular localization dynamics of the aggregation-prone LINE-1 ORF1p RNA-binding protein. However, we failed to find compelling evidence for misregulation of LINE-1 retrotransposons in sporadic ALS nor a clear effect of ALS-associated TDP-43 protein on L1 expression. In sum, our study reveals that the interplay of active retrotransposons and the molecular features of ALS are more complex than anticipated. Thus, the potential consequences of altered retrotransposon activity for ALS and other neurodegenerative disorders are worthy of continued investigation.


Monte Carlo characterizations mapping of the (γ,n) and (n,γ) photonuclear reactions in the high energy X-ray radiation therapy.
The aim of this work was to map the characteristics of (n,γ) and (γ,n) reactions in a high energy photon radiation therapy. Photoneutrons produced in the high energy X-Ray radiation therapy may damage patients and staff. It is due to high RBE of the produced neutrons according to their energy and isotropic emission. Characterization of the photoneutrons can help us in appropriate shielding. This study focused on the photoneutron and capture gamma ray phenomena. Characteristics such as dose value, fluence and spectra of both the neutrons and the by produced prompt gamma ray were described. Neutron and prompt gamma spectra in different points showed the neutrons to be thermalized when increasing the distance from the linac. Energy of the neutrons changed from about 0.6 MeV at the isocentre to around 10(-08) MeV at the outer door position. Although the neutrons were found as fast neutrons, their spectra showed they were thermal neutrons at the outer door position. Additionally, it was seen that the energy of the gamma rays is higher than the scattered X-ray energy. The energy of gamma rays was seen to be up to 10 MeV while the linac photons had energy lower than 1 MeV. Neutron source strength obtained in this work was in good agreement with the published data, which may be a confirmation of our simulation accuracy. The study showed that the Monte Carlo simulation can be applied in the radiotherapy and industrial radiation works as a useful and precise estimator. We also concluded that the dose from the prompt gamma ray at the outer door location is higher than the scattered radiation from the linac and should be considered in the shielding.


Epicardial Adipose Tissue: Another Tassel in the Complex Fabric of Atherosclerosis.
Atherosclerosis affects the majority of adult individuals in industrialized nations and it is beginning to affect even traditionally spared populations. The classic view has been that the precipitating events are intraluminal. However, good evidence supports the possibility that at least part of the atherosclerosis burden may be the consequence of extra-luminal noxious stimuli. Additionally, the epidemic of obesity, insulin resistance and diabetes mellitus has generated a strong interest in the potential role of visceral adipose tissue as an extra-luminal promoter of atherosclerosis. The epicardial space is filled with adipose tissue with an embryological origin similar to that of abdominal visceral fat. Both fats are highly inflamed in obese patients, patients with the metabolic syndrome and in those with established coronary artery disease. Additionally they are capable of secreting large quantities of pro-inflammatory cytokines and free fatty acids but also anti-inflammatory adipokines like adiponectin. In this manuscript we review the current evidence supporting the role of epicardial adipose tissue in the development of atherosclerosis and its complications.


[Sex difference of functional and structural alterations in the myocardium of rats with hypothyroidism].
In experiments on sexually mature rats we studied specific cholinergic regulations of the heart and the degree of its structural damage in hypothyroidism, depending on gender and hormone-productive activity of the gonads. Hypothyroidism in sexually mature males and females was modelled with mercazolil intragastric administration (75 mg/kg) daily during 15 days. We also studied the intensity of bradycardia, which occurred in response to electrical stimulation of vagus nerve and intravenous acetylcholine administration. The degree of structural heart damage was assessed by the percentage of damaged cardiomyocytes in the ventricles of myocardium. It was found that one of the mechanisms of bradycardia in merkazolil-induced hypothyroidism is an increase of the sensitivity of sinus node cholinergic receptors and release of more quanta of acetylcholine from stimulated nerves vagus endings, what was more intense in females. The intensity of bradycardia in hypothyroidism was more significant in gonadectomized animals than in individuals with preserved gonads. The mechanisms of its occurrence in males consist of release of greater amount of acetylcholine from the endings of the nerves vagus, and in females it was the result of significant increase of the sensitivity of sinus node cholinergic receptors. Regardless of the gonads activity, structural damage of the myocardium in merkazolil-induced hypothyroidism was more intensive in female rats.


Perspectives on molecular and cellular exercise physiology.
A challenge to applied physiologists is to continue to apply new methods to their field. The purpose of this review is to speculate how recent advances in the fields of molecular and cell biology might be applied to exercise physiology to provide greater insights into the mechanisms underlying adaptations to a single bout of exercise and to repeated bouts of exercise (training). The review is organized to consider the actions occurring outside the cell, through the cellular membrane and cytoplasm to the nucleus. These topics are as follows: blood-borne signals, polypeptide growth factors, membrane receptors (insulin receptor, adrenergic receptors, acetylcholine receptors, phosphorylation of receptors, and mutant receptors), transduction through the plasmalemma (glucose transporters and G proteins), second messengers (phosphatidylinositol phosphates and calcium), cis sequences required for muscle-specific gene expression, transcriptional regulation by physiological signals, and gene expression. Exercise physiology is a challenging discipline that integrates molecules, cell-to-cell interaction, tissues, and the whole organism during the physiological stress of exercise by the live, unanesthetized animal.


Activation of the Intrinsic Pain Inhibitory Circuit from the Midcingulate Cg2 to Zona Incerta Alleviates Neuropathic Pain.
Neuropathic pain is one of the most common and notorious neurological diseases. The changes in cerebral structures after nerve injury and the corresponding contributions to neuropathic pain are not well understood. Here we found that the majority of glutamatergic neurons in the area 2 of midcingulate cortex (MCC Cg2Glu) were inhibited by painful stimulation in male mice. Optogenetic manipulation revealed that these neurons were tonically involved in the inhibitory modulation of multimodal nociception. We further identified the projections to GABAergic neurons in the zona incerta (ZIGABA) mediated the pain inhibitory role. However, MCC Cg2Glu became hypoactive after nerve injury. Although a brief activation of the MCC Cg2Glu to ZIGABA circuit was able to relieve the aversiveness associated with spontaneous ongoing pain, consecutive activation of the circuit was required to alleviate neuropathic allodynia. In contrast, glutamatergic neurons in the area 1 of MCC played opposite roles in pain modulation. They became hyperactive after nerve injury and only consecutive inhibition of their activity relieved allodynia. These results demonstrate that MCC Cg2Glu constitute a component of intrinsic pain inhibitory circuitry and their hypoactivity underlies neuropathic pain. We propose that selective and persistent activation of the MCC Cg2Glu to ZIGABA circuit may serve as a potential therapeutic strategy for this disease.SIGNIFICANCE STATEMENT Glutamatergic neurons in the area 2 of midcingulate cortex (MCC Cg2Glu) are tonically involved in the intrinsic pain inhibition via projecting to GABAergic neurons in the zona incerta. They are hypoactive after nerve injury. Selective activation of the circuit compensates the reduction of its analgesic strength and relieves neuropathic pain. Therefore, MCC Cg2Glu and the related analgesic circuit may serve as therapeutic targets for neuropathic pain. In contrast, MCC Cg1Glu have an opposite role in pain modulation and become hyperactive after nerve injury. The present study provides novel evidence for the concept that neuropathic pain is associated with the dysfunction of endogenous pain modulatory system and new perspective on the treatment of neuropathic pain.


Ledipasvir/sofosbuvir for treatment of hepatitis C virus in sofosbuvir-experienced, NS5A treatment-naïve patients: Findings from two randomized trials.
We report data from two similarly designed studies that evaluated the efficacy, safety, and optimal duration of ledipasvir/sofosbuvir (LDV/SOF) ± ribavirin (RBV) for retreatment of chronic hepatitis C virus (HCV) in individuals who failed to achieve sustained virological response (SVR) with prior SOF-based, non-NS5A inhibitor-containing regimens. The RESCUE study enrolled HCV mono-infected adults with genotype (GT) 1 or 4. Non-cirrhotic participants were randomized to 12 weeks of LDV/SOF or LDV/SOF + RBV. Compensated cirrhotic participants were randomized to LDV/SOF + RBV (12 weeks) or LDV/SOF (24 weeks). The AIDS Clinical Trials Group A5348 study randomized genotype 1 adults with HCV/HIV co-infection to LDV/SOF + RBV (12 weeks) or LDV/SOF (24 weeks). Both studies used SVR at 12 weeks post-treatment (SVR12) as the primary endpoint. In the RESCUE study, 82 participants were randomized and treated, and all completed treatment. Overall, SVR12 was 88% (72/82); 81-100% in non-cirrhotic participants treated with LDV/SOF or LDV/SOF + RBV for 12 weeks and 80-92% in cirrhotic participants treated with LDV/SOF + RBV for 12 weeks or LDV/SOF for 24 weeks. Adverse events (AEs), mostly mild-to-moderate in severity, were experienced by 78% of participants, with headache and fatigue most frequently reported. One serious AE, not related to treatment, was observed. No premature discontinuations of study drug, or deaths occurred. In the A5348 study, seven participants were randomized (cirrhotic n = 1; GT1a n = 5) and all attained SVR12, with no serious AEs or premature discontinuations. In this SOF-experienced, NS5A inhibitor-naïve population, which included participants with cirrhosis or HCV/HIV co-infection, high SVR12 rates were achieved.


Management of travelers with fever and exanthema, notably dengue and chikungunya infections.
Febrile exanthema frequently presents as an acute condition in travelers. Sixty-two travelers who presented with febrile exanthema were prospectively included over a 20-month period. Diagnostic tests were performed according to clinical presentation and risk exposures. Symptoms occurred after return in 56% of these travelers, and the median interval between return and symptom onset was 2 days. The 3 main travel destinations were the Indian Ocean (35%), Africa (21%), and Asia (18%). The 3 main etiologies were chikungunya (35%), dengue (26%), and African tick bite fever (ATBF) (10%). Travel to the Indian Ocean and South Africa was significantly associated with respectively chikungunya and ATBF. Arthralgias were significantly more frequent in chikungunya than in dengue, whereas leucopenia, neutropenia, and thrombopenia were significantly more frequent in dengue. Travelers presenting with febrile exanthema should therefore be screened for arboviral infections according to the area visited.


Dehydration of Seeds in Intact Tomato Fruits.
Although the moisture content of the placenta and pericarp of tomato fruits older than 15 days never dropped below about 94 percent, the seeds dehydrated to approximately 50 percent moisture as these organs developed and matured. There was a loss in the total amount, as well as the percentage, of water per seed. Part of this dehydration was mediated by the seeds themselves and part appeared to be due to other portions of the fruit.


Evolution of polyploidy and the diversification of plant-pollinator interactions.
One of the major mechanisms of plant diversification has been the evolution of polyploid populations that differ from their diploid progenitors in morphology, physiology, and environmental tolerances. Recent studies have indicated that polyploidy may also have major effects on ecological interactions with herbivores and pollinators. We evaluated pollination of sympatric diploid and tetraploid plants of the rhizomatous herb Heuchera grossulariifolia (Saxifragaceae) along the Selway and Salmon Rivers of northern Idaho, USA, during four consecutive years. Previous molecular and ecological analyses had indicated that the tetraploid populations along these two river systems are independently derived and differ from each other in multiple traits. In each region, we evaluated floral visitation rate by all insect visitors, pollination efficacy of all major visitors, and relative contribution of all major pollinators to seed set. In both regions, diploid and tetraploid plants attracted different suites of floral visitors. Most pollination was attributable to several bee species and the moth Greya politella. Lasioglossum bees preferentially visited diploid plants at Lower Salmon but not at Upper Selway, queen Bombus centralis preferentially visited tetraploids at both sites, and worker B. centralis differed between sites in their cytotype preference. Hence, diploid and autotetraploid H. grossulariifolia plants act essentially as separate ecological species and may experience partial reproductive isolation through differential visitation and pollination by their major floral visitors. Overall the results, together with recent results from other studies, suggest that the repeated evolution of polyploidy in plants may contribute importantly to the structure and diversification of ecological interactions in terrestrial communities.


How is research publishing going to progress in the next 20 years?: transcription of session for editors, associate editors, publishers and others with an interest in scientific publishing held at IADR meeting in Seattle on Wednesday, 20 March 2013.
On March 20th 2013, a one-hour session for Editors, Associate Editors, Publishers and others with an interest in scientific publishing was held at the IADR International Session in Seattle. Organised by Kenneth Eaton and Christopher Lynch (Chair and Secretary, respectively, of the British Dental Editors Forum), the meeting sought to bring together leading international experts in dental publishing, as well as authors, reviewers and students engaged in research. The meeting was an overwhelming success, with more than 100 attendees. A panel involving four leading dental editors led a discussion on anticipated developments in publishing dental research with much involvement and contribution from audience members. This was the third such meeting held at the IADR for Editors, Associate Editors, Publishers and others with an interest in scientific publishing. A follow-up session will take place in Cape Town on 25 June 2014 as part of the annual IADR meeting. The transcript of the Seattle meeting is reproduced in this article. Where possible speakers are identified by name. At the first time of mention their role/position is also stated, thereafter only their name appears. We are grateful to Stephen Hancocks Ltd. for their generous sponsorship of this event. For those who were not able to attend the authors hope this article gives a flavour of the discussions and will encourage colleagues to attend future events. Involvement is open to Editors, Associate Editors, Publishers and others with an interest in scientific publishing. It is a very open group and all those with an interest will be welcome to join in.


Effectiveness of physical activity interventions in achieving behaviour change maintenance in young and middle aged adults: A systematic review and meta-analysis.
Physical activity (PA) interventions are generally effective in supporting short-term behaviour change, but increases are not always maintained. This review examined the effectiveness of PA interventions for behaviour change maintenance in young and middle-aged adults, and investigated which Behaviour Change Techniques (BCTs) and other intervention features were associated with maintenance. Six databases (Medline, EMBASE, PsycINFO, Cochrane Database of Systematic Reviews, CINAHL, Web of Science) were systematically searched. Eligibility criteria were controlled trials investigating the effectiveness of PA interventions with adult (mean age 18-64 years) non-clinical populations using validated measures of PA behaviour at baseline and ≥six months' post-baseline. Results were pooled in meta-analyses using standardised mean differences (SMD) at five time intervals (6-9, 9-15, 15-21, 21-24, >24 months). Moderator analyses investigated the influence of sample and intervention characteristics on PA maintenance at 6-9 months. Sixty-two studies were included. PA interventions had a significant effect on behaviour maintenance 6-15 months post-baseline relative to controls. Interventions had a larger effect on maintenance at 6-9 months (SMD = 0.28; 95% CI: 0.20, 0.35; I2 = 73%) compared to 9-15 months (SMD = 0.20; 95% CI: 0.13, 0.26; I2 = 70%). Beyond 15 months, PA measurements were infrequent with little evidence supporting maintenance. Moderator analyses showed some BCTs and intervention settings moderated PA outcomes at 6-9 months. A multivariable meta-regression model showed interventions using the BCTs 'Prompt self-monitoring of behavioural outcome' (b = 1.46, p < 0.01) and 'Use of follow-up prompts' (b = 0.38, p < 0.01) demonstrated greater effectiveness at promoting PA maintenance at 6-9 months. Interventions implemented in primary care (versus community or workplace/university) settings (b = -0.13, p = 0.10) tended to demonstrate less effectiveness. This review provides evidence of some effective BCTs for maintaining behaviour to 15 months. Greater consideration must be given to how future interventions encourage and measure maintenance of changes, and investigate broader psychological, social and environmental influences of PA behaviour. PROSPERO 2015:CRD42015025462.


Frequent emergence of porin-deficient subpopulations with reduced carbapenem susceptibility in ESBL-producing Escherichia coli during exposure to ertapenem in an in vitro pharmacokinetic model.
Ertapenem resistance is increasing in Enterobacteriaceae. The production of extended-spectrum β-lactamases (ESBLs) and reduced expression of outer membrane porins are major mechanisms of resistance in ertapenem-resistant Klebsiella pneumoniae. Less is known of ertapenem resistance in Escherichia coli. The aim of this study was to explore the impact of ESBL production in E. coli on the antibacterial activity of ertapenem. Two E. coli strains, with and without ESBL production, were exposed to ertapenem in vitro for 48 h at concentrations simulating human pharmacokinetics with conventional and higher dosages. Isolates with non-susceptibility to ertapenem (MICs 0.75-1.5 mg/L) were detected after five of nine time-kill experiments with the ESBL-producing strain. All of these isolates had ompR mutations, which reduce the expression of outer membrane porins OmpF and OmpC. Higher dosage did not prevent selection of porin-deficient subpopulations. No mutants were detected after experiments with the non-ESBL-producing strain. Compared with other experiments, experiments with ompR mutants detected in endpoint samples showed significantly less bacterial killing after the second dose of ertapenem. Impaired antibacterial activity against E. coli with ESBL production and ompR mutation was also demonstrated in time-kill experiments with static antibiotic concentrations. The combination of ESBL production and porin loss in E. coli can result in reduced susceptibility to ertapenem. Porin-deficient subpopulations frequently emerged in ESBL-producing E. coli during exposure to ertapenem at concentrations simulating human pharmacokinetics. Inappropriate use of ertapenem should be avoided to minimize the risk of selection of ESBL-producing bacteria with reduced susceptibility to carbapenems.


[Rethoracotomy because of hemorrhage in the early postoperative period after operations on the lungs and mediastinal organs].
During the period from 1967 till 1994 2760 operations were performed on the lungs and mediastinum organs. In 75 patients (2,72%) intrapleural bleedings took place at the early postoperative period which led to rethoracotomies. Based on an analysis of their material the authors made a conclusion that in all the cases in which the intrapleural bleeding was diagnosed rethoracotomy was necessary. All the rethoracotomies were divided into urgent which should be performed as soon as diagnosis of continuing massive intrapleural bleeding was made, and postponed ones when the bleeding was not threatening during the first hours after operation, AP did not decrease, there was no considerable drop of Hb and Ht. From the 75 patients ten patients died after rethoracotomy and 65 patients recovered.


[Medulloblastomas: clinical, epidemiological and pathological findings in 28 cases].
We report the clinical, epidemiological and pathological findings of 28 patients with medulloblastoma: 22 were male; age ranged from 1 to 50 years, with a mean of 15 years. The most frequent symptoms and signs were headache (64%) and vomiting (64%). Only one patient showed a desmoplastic medulloblastoma variant, the others showed classical medulloblastomas. Regarding treatment, most patients were submitted to total resection (n=10) or partial tumorectomy (n=7). A low rate of tumoral recurrence was observed, occurring in 21% of the patients (n=6) after 4 years of follow-up. Chemotherapy seemed to contribute to a lower recurrence rate amongst our patients. Our findings are similar to those reported in literature, thus helping to understand the biological behavior of this type of tumor.


Effect of chronic treatment with atenolol and prazosin in streptozotocin induced diabetic rats.
Diabetes-mellitus was induced in rats by single intravenous injection of (45 mg/kg) streptozotocin (STZ). STZ diabetic rats showed hypertension, decreased cardiac functions, cardiomyopathy and hypercholesterolemia observed at the end of six weeks. Chronic treatment with atenolol (10 mg/kg) for six weeks in the diabetic rats reduced the elevated blood pressure, but failed to prevent STZ induced other complications. Chronic treatment with prazosin (1 mg/kg, po) in the diabetic rats, reduced the elevated blood pressure and also partially prevented hypercholesterolemia, cardiac dysfunctions and in particular the cardiomyopathy. The results suggest that prazosin may be a better option as compared to atenolol in hypertension when it is associated with diabetes mellitus.


[Regulation of free radical oxidation of pulmonary surfactant lipids].
Surface-active substances of lung tissue, covering the alveoli, resembled the biological membranes in their structure. Their composition was controlled, among the other factors, also by free radical oxidation reactions. Under conditions of circulatory hypoxia the activity of antioxidative enzymes correlated with the alterations in antioxidative activity, with the rates of spontaneous and inducible lipid peroxidation in lung surfactants. As surfactant exhibited a limited antioxidative activity important for resistance of alveolar pneumocytes, their functions may be controlled by various exogenous antioxidants.


Clinical impacts of additive use of olmesartan in hypertensive patients with chronic heart failure: the supplemental benefit of an angiotensin receptor blocker in hypertensive patients with stable heart failure using olmesartan (SUPPORT) trial.
We examined whether an additive treatment with an angiotensin receptor blocker, olmesartan, reduces the mortality and morbidity in hypertensive patients with chronic heart failure (CHF) treated with angiotensin-converting enzyme (ACE) inhibitors, β-blockers, or both. In this prospective, randomized, open-label, blinded endpoint study, a total of 1147 hypertensive patients with symptomatic CHF (mean age 66 years, 75% male) were randomized to the addition of olmesartan (n = 578) to baseline therapy vs. control (n = 569). The primary endpoint was a composite of all-cause death, non-fatal acute myocardial infarction, non-fatal stroke, and hospitalization for worsening heart failure. During a median follow-up of 4.4 years, the primary endpoint occurred in 192 patients (33.2%) in the olmesartan group and in 166 patients (29.2%) in the control group [hazard ratio (HR) 1.18; 95% confidence interval (CI), 0.96-1.46, P = 0.112], while renal dysfunction developed more frequently in the olmesartan group (16.8 vs. 10.7%, HR 1.64; 95% CI 1.19-2.26, P = 0.003). Subgroup analysis revealed that addition of olmesartan to combination of ACE inhibitors and β-blockers was associated with increased incidence of the primary endpoint (38.1 vs. 28.2%, HR 1.47; 95% CI 1.11-1.95, P = 0.006), all-cause death (19.4 vs. 13.5%, HR 1.50; 95% CI 1.01-2.23, P = 0.046), and renal dysfunction (21.1 vs. 12.5%, HR 1.85; 95% CI 1.24-2.76, P = 0.003). Additive use of olmesartan did not improve clinical outcomes but worsened renal function in hypertensive CHF patients treated with evidence-based medications. Particularly, the triple combination therapy with olmesartan, ACE inhibitors and β-blockers was associated with increased adverse cardiac events. This study is registered at clinicaltrials.gov-NCT00417222.


Hand strengths in carpal tunnel syndrome.
The purpose of this study was to determine whether a measurable decrease in isokinetic (dynamic) and isometric (static) hand strengths occurs in carpal tunnel syndrome (CTS) patients. Eighteen CTS patients and 20 healthy controls were included in the study. Isokinetic (eccentric and concentric) and isometric grip and pinch strengths were measured with a Biodex System 3 dynamometer (Biodex Medical System, Inc. New York). All strength measurements, except isometric and isokinetic (concentric/eccentric) three-point pinch and isokinetic (concentric) tip pinch, revealed statistically significant differences between CTS patients and controls. Measurable decrease in hand strengths may exist in CTS despite normal manual assessments. Although both isokinetic (dynamic) and isometric (static) dynamometers are capable of detecting this decrease, neither technique seems better than the other.


Chiral recognition by nucleosides and nucleotides: resolution of helicenes by high-performance liquid chromatography.
Chiral recognition by nucleosides and nucleotides coated on silica gel was studied by high-performance liquid chromatography. Helicenes, which are chiral polyaromatic hydrocarbons, were used as probes. Stereoselectivity was detected when the nucleobase was a purine (adenosine 3', 5'-monophosphate, an guanosine), but was not detected with the pyrimidine derivative uridine. For a given nucleobase (adenine), all changes in the ribose moiety affected the resolution factors, which ranged between 1.03 and 1.074. These results might be relevant to the enantioselectivity of carcinogenic metabolites of polyaromatic hydrocarbons.


Visual- and saccade-related signals in the primate inferior colliculus.
The inferior colliculus (IC) is normally thought of as a predominantly auditory structure because of its early position in the ascending auditory pathway just before the auditory thalamus. Here, we show that a majority of IC neurons (64% of 180 neurons) in awake monkeys carry visual- and/or saccade-related signals in addition to their auditory responses (P < 0.05). The response patterns involve primarily excitatory visual responses, but also increased activity time-locked to the saccade, slow rises in activity time-locked to the onset of the visual stimulus, and inhibitory responses. The presence of these visual-related signals suggests that the IC plays a role in integrating visual and auditory information. More broadly, our results show that interactions between sensory pathways can occur at very early points in sensory processing streams, which implies that multisensory integration may be a low-level rather than an exclusively high-level process.


Confocal microwave imaging for breast cancer detection: delay-multiply-and-sum image reconstruction algorithm.
A new image reconstruction algorithm, termed as delay-multiply-and-sum (DMAS), for breast cancer detection using an ultra-wideband confocal microwave imaging technique is proposed. In DMAS algorithm, the backscattered signals received from numerical breast phantoms simulated using the finite-difference time-domain method are time shifted, multiplied in pair, and the products are summed to form a synthetic focal point. The effectiveness of the DMAS algorithm is shown by applying it to backscattered signals received from a variety of numerical breast phantoms. The reconstructed images illustrate improvement in identification of embedded malignant tumors over the delay-and-sum algorithm. Successful detection and localization of tumors as small as 2 mm in diameter are also demonstrated.


Effect of inherited abnormalities of calcium regulation on human neuromuscular transmission.
Synaptotagmins are abundant synaptic proteins that represent the best candidate for the calcium sensor at the nerve terminal. The pore-forming, voltage-sensing transmembrane alpha-1 subunit of the P/Q voltage-gated calcium channel (or Ca(v)2.1) encoded by the CACNA1A gene is another major component of the process of action potential-evoked exocytosis at the adult mammalian neuromuscular junction. Defects of these proteins, in nonhuman species, result in severe disruption of rapid synaptic transmission. This paper investigates the molecular bases of inherited presynaptic deficits of neuromuscular transmission in humans. Patients with congenital presynaptic failure, including two patients with episodic ataxia type 2 (EA-2) due to CACNA1A mutations, were studied with muscle biopsy, microelectrode studies, electron microscopy, DNA amplification, and sequencing. All patients, including EA-2 patients, showed selective failure of the action potential-dependent release without reduction of the spontaneous release of neurotransmitter. In addition, patients with EA-2 showed partial blockade of neuromuscular transmission with the N-type blocker omega-conotoxin not seen in controls. The EM showed a varied degree of increased complexity of postsynaptic folds. Mutational analysis in candidate genes, including human synaptotagmin II, syntaxin 1A, synaptobrevin I, SNAP 25, CACNA1A, CACNB2, and Rab3A, was unrevealing. Although no mutations in candidate genes were found in patients with inborn presynaptic failure, functional and structural similarities between this group and patients with EA-2 due to CACNA1A mutations suggest a common pathogenic mechanism.


Food safety in the domestic environment: the effect of consumer risk information on human disease risks.
The improvement of food safety in the domestic environment requires a transdisciplinary approach, involving interaction between both the social and natural sciences. This approach is applied in a study on risks associated with Campylobacter on broiler meat. First, some web-based information interventions were designed and tested on participant motivation and intentions to cook more safely. Based on these self-reported measures, the intervention supported by the emotion "disgust" was selected as the most promising information intervention. Its effect on microbial cross-contamination was tested by recruiting a set of participants who prepared a salad with chicken breast fillet carrying a known amount of tracer bacteria. The amount of tracer that could be recovered from the salad revealed the transfer and survival of Campylobacter and was used as a measure of hygiene. This was introduced into an existing risk model on Campylobacter in the Netherlands to assess the effect of the information intervention both at the level of exposure and the level of human disease risk. We showed that the information intervention supported by the emotion "disgust" alone had no measurable effect on the health risk. However, when a behavioral cue was embedded within the instruction for the salad preparation, the risk decreased sharply. It is shown that a transdisciplinary approach, involving research on risk perception, microbiology, and risk assessment, is successful in evaluating the efficacy of an information intervention in terms of human health risks. The approach offers a novel tool for science-based risk management in the area of food safety.


[Interhemispheric motor asymmetry of conditioned feedback].
The interhemispheric asymmetry and lateralization of motor functions, as applied to backward conditioning, was studied in two dogs that afterwards turned to be a right-hander and a left-hander. In the process of food-procuring instrumental conditioning it was revealed that one of the dogs predominantly used the right paw (with the dominance of the left hemisphere) and the second animal used the left paw (the right brain hemisphere dominated). The earliest signs of lateralization of the motor functions were observed in the backward conditioned connections (which had been formed earlier than the direct connections or direct conditioned reflexes). Consequently, the presence and sign of asymmetry can be predicted on the basis of manifestation of the backward connections observed at the early stages of acquisition of food-procuring behavior. The backward connections are strictly specific and have the same lateralization as the direct ones.


A novel approach to identify the miRNA-mRNA causal regulatory modules in Cancer.
MicroRNAs (miRNAs) play an essential role in many biological processes by regulating the target genes, especially in the initiation and development of cancers. Therefore, the identification of the miRNA-mRNA regulatory modules is important for understanding the regulatory mechanisms. Most computational methods only used statistical correlations in predicting miRNA-mRNA modules, and neglected the fact there are causal relationships between miRNAs and their target genes. In this paper, we propose a novel approach called CALM(the causal regulatory modules) to identify the miRNA-mRNA regulatory modules through integrating the causal interactions and statistical correlations between the miRNAs and their target genes. Our algorithm largely consists of three steps: it first forms the causal regulatory relationships of miRNAs and genes from gene expression profiles and detects the miRNA clusters according to the GO function information of their target genes, then expands each miRNA cluster by greedy adding(discarding) the target genes to maximize the modularity score. To show the performance of our method, we apply CALM on four datasets including EMT, breast, ovarian, thyroid cancer and validate our results. The experiment results show that our method can not only outperform the compared method, but also achieve ideal overall performance in terms of the functional enrichment.


Immunolocalization of estrogen-producing and metabolizing enzymes in benign breast disease: comparison with normal breast and breast carcinoma.
It is well known that estrogens play important roles in the cell proliferation of breast carcinoma. Benign breast disease (BBD) contains a wide spectrum of diseases, and some are considered an important risk factor for subsequent breast carcinoma development. However, the significance of estrogens in BBD has remained largely unknown. Therefore, in this study, we examined tissue concentrations of estrogens and immunolocalization of estrogen-producing/metabolizing enzymes in BBD, and compared these findings with those in the normal breast and ductal carcinoma in situ (DCIS). Tissue concentration of estradiol in BBD (n = 9) was significantly (3.4-fold) higher than normal breast (n = 9) and nearly the same (0.7-fold) as in DCIS (n = 9). Immunoreactivity of estrogen sulfotransferase in BBD was significantly lower (n = 82) than normal breast (n = 28) but was not significantly different from DCIS (n = 28). Aromatase and steroid sulfatase immunoreactivities tended to be higher (P = 0.07) in BBD than in normal breast, and 17β-hydroxysteroid dehydrogenase type 1 immunoreactivity was significantly higher in BBD than normal breast in the postmenopausal tissues. Immunoreactivity of estrogen and progesterone receptors was also significantly higher in BBD than normal breast. These results suggest that tissue concentration of estradiol is increased in BBD at a level similar to DCIS, which is considered mainly due to loss of estrogen sulfotransferase expression. Increased local estradiol concentration in BBD due to aberrant expression of estrogen-producing/metabolizing enzymes may play important roles in the accumulation of estradiol-mediated growth and/or subsequent development of breast carcinoma.


Role of individual surface charges of voltage-gated K channels.
Fixed charges on the extracellular surface of voltage-gated ion channels influence the gating. In previous studies of cloned voltage-gated K channels, we found evidence that the functional surface charges are located on the peptide loop between the fifth transmembrane segment and the pore region (the S5-P loop). In the present study, we determine the role of individual charges of the S5-P loop by correlating primary structure with experimentally calculated surface potentials of the previously investigated channels. The results suggest that contributions to the surface potential at the voltage sensor of the different residues varies in an oscillating pattern, with the first residue of the N-terminal end of the S5-P loop, an absolutely conserved glutamate, contributing most. An analysis yields estimates of the distance between the residues and the voltage sensor, the first N-terminal residue being located at a distance of 5-6 A. To explain the results, a structural hypothesis, comprising an alpha-helical N-terminal end of the S5-P loop, is presented.


Learning to Let Go: A Cognitive-Behavioral Model of How Psychedelic Therapy Promotes Acceptance.
The efficacy of psychedelic-assisted therapies for mental disorders has been attributed to the lasting change from experiential avoidance to acceptance that these treatments appear to facilitate. This article presents a conceptual model that specifies potential psychological mechanisms underlying such change, and that shows substantial parallels between psychedelic therapy and cognitive behavioral therapy: We propose that in the carefully controlled context of psychedelic therapy as applied in contemporary clinical research, psychedelic-induced belief relaxation can increase motivation for acceptance via operant conditioning, thus engendering episodes of relatively avoidance-free exposure to greatly intensified private events. Under these unique learning conditions, relaxed avoidance-related beliefs can be exposed to corrective information and become revised accordingly, which may explain long-term increases in acceptance and corresponding reductions in psychopathology. Open research questions and implications for clinical practice are discussed.


Epithelial expression of mRNA and protein for IL-6, IL-10 and TNF-alpha in endobronchial biopsies in horses with recurrent airway obstruction.
The aim of this study was to evaluate the contribution of bronchial epithelium to airway inflammation, with focus on mRNA and protein expression of cytokines of innate immunity IL-6, IL-10 and TNF-alpha, in horses with Recurrent Airway Obstruction (RAO) during exacerbation and in remission. Despite marked clinical and physiologic alterations between exacerbation and after remission in the RAO horses no differences were detected in either cytokine mRNA or protein levels. Moreover, the expression of investigated cytokines in RAO horses on pasture did not differ from controls. In comparing real-time PCR analysis to results of immunohistochemistry only IL-10 mRNA and protein levels in RAO horses on pasture were significantly correlated (rs = 0.893, p = 0.007). Curiously, in controls examined on pasture the TNF-alpha protein level was positively correlated to IL-10 mRNA expression (rs = 0.967, p = 0.007) and negatively correlated to IL-6 mRNA expression (rs = -0.971, p = 0.001). Given the complementary relationship of assessing cytokines directly by immunohistochemistry, or indirectly by PCR to mRNA, the lack of significant changes in either mRNA or protein levels of IL-6, IL-10 or TNF-alpha mRNA in RAO horses in exacerbation suggests that these particular cytokines in bronchial tissue may not play a substantive role in the active inflammation of this disease. To support this contention further studies examining time dependency of expression of IL-6, IL-10 or TNF-alpha are needed, as is expansion of the range of cytokines to include other key regulators of airway inflammation.


Activation of ExoU phospholipase activity requires specific C-terminal regions.
Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen that utilizes a type III secretion system to subvert host innate immunity. Of the 4 known effector proteins injected into eukaryotic cells, ExoS and ExoU are cytotoxic. The cytotoxic phenotype of ExoU depends on the enzymatic activity of the patatin-like phospholipase A(2) domain localized to the N-terminal half of the protein. Amino acid residues located within the C-terminal region of ExoU are postulated to be required for trafficking or localization to the plasma membrane of eukaryotic cells. This report describes the characterization of a transposon-based linker insertion library in ExoU. Utilizing an unbiased screening approach and sensitive methods for measuring enzymatic activity, we identified regions of ExoU that are critical for activation of the phospholipase activity by the only known cofactor, SOD1. Insertions at D572 and L618 reduced the rate of substrate cleavage. Enzymatic activity could be restored to almost parental levels when SOD1 concentrations were increased, suggesting that the linker insertion disrupted the interaction between ExoU and SOD1. An enzyme-linked immunosorbent assay (ELISA)-based binding test was developed to measure ExoU-SOD1 binding. These experiments suggest that ExoU activation by SOD1 is hampered by linker insertion. ExoU derivatives harboring minimal phospholipase activity retained biological activity in tissue culture assays. These proteins affected primarily cellular architecture in a manner similar to that of ExoT. Our studies suggest that conformational changes in ExoU are facilitated by SOD1. Importantly, the level of phospholipase activity influences the biological outcome of ExoU intoxication.


Mechanism of actin-dependent activation of nucleotidyl cyclase toxins from bacterial human pathogens.
Bacterial human pathogens secrete initially inactive nucleotidyl cyclases that become potent enzymes by binding to actin inside eukaryotic host cells. The underlying molecular mechanism of this activation is, however, unclear. Here, we report structures of ExoY from Pseudomonas aeruginosa and Vibrio vulnificus bound to their corresponding activators F-actin and profilin-G-actin. The structures reveal that in contrast to the apo-state, two flexible regions become ordered and interact strongly with actin. The specific stabilization of these regions results in an allosteric stabilization of the nucleotide binding pocket and thereby to an activation of the enzyme. Differences in the sequence and conformation of the actin-binding regions are responsible for the selective binding to either F- or G-actin. Other nucleotidyl cyclase toxins that bind to calmodulin rather than actin undergo a similar disordered-to-ordered transition during activation, suggesting that the allosteric activation-by-stabilization mechanism of ExoY is conserved in these enzymes, albeit the different activator.


Preparation and purification of polymerized actin from sea urchin egg extracts.
Isotonic extracts of the soluble cytoplasmic proteins of sea urchin eggs, containing sufficient EGTA to reduce the calcium concentration to low levels, form a dense gel on warming to 35-40 degrees C. Although this procedure is similar to that used to polymerize tubulin from mammalian brain, sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows this gel to have actin as a major component and to contain no tubulin. If such extracts are dialyzed against dilute salt solution, they no longer respond to warming, but gelation will occur if they are supplemented with 1 mM ATP and 0.020 M KCl before heating. Gelation is not temperature reversible, but the gelled material can be dissolved in 0.6-1 M KCl and these solutions contain F-actin filaments. These filaments slowly aggregate to microscopic, birefringent fibrils when 1 mM ATP is added to the solution, and this procedure provides a simple method for preparing purified actin. the supernate remaining after actin removal contains the other two components of the gel, proteins of approximately 58,000 and 220,000 mol wt. These two proteins plus actin recombine to form the original gel material when the ionic strength is reduced. This reaction is reversible at 0 degrees C, and no heating is required.


Occurrence of nitrosamines and their precursors in drinking water systems around mainland China.
N-Nitrosamines (NAs) in drinking water have attracted considerable attention in recent years due to their high carcinogenicity, frequent occurrence, and their potential regulation. During the past three years, we have collected about 164 water samples of finished water, tap water, and source water from 23 provinces, 44 cities from large cities to small towns, and 155 sampling points all over China. The occurrence of NAs in the finished and tap water was much higher in China than that in the U.S. Nine NAs were measured and NDMA had the highest concentration. The occurrence of NDMA was in 33% of the finished waters of water treatment plants and in 41% of the tap waters. The average NDMA concentration in finished and tap waters was 11 and 13 ng/L, respectively. Formation potentials (FPs) of source waters were examined with an average NDMA FP of 66 ng/L. Large variations in NA occurrence were observed geographically in China and temporally in different seasons. The Yangtze River Delta area, one sub-area in East China, had the highest concentrations of NAs, where the average NDMA concentrations in the finished and tap water were 27 and 28.5 ng/L, respectively, and the average NDMA FP in the source water was 204 ng/L. NA control may be achieved by applying breakpoint free chlorination and/or advanced treatment of ozone - granular activated carbon process to remove the NA precursors before disinfection.


Pharmacokinetics of ethylene glycol. I. Plasma disposition after single intravenous, peroral, or percutaneous doses in female Sprague-Dawley rats and CD-1 mice.
The pharmacokinetics of [1,2-14C]ethylene glycol (EG) were evaluated in female Sprague-Dawley rats and CD-1 mice to characterize the plasma disposition after intravenous (IV), peroral (PO), and percutaneous (PC) doses. Rats were given doses of 10 or 1000 mg/kg by each route, and additional PO doses of 400, 600, or 800 mg/kg. Mice were also given IV and PO (bolus gavage) doses of 10 or 1000 mg/kg, and additional PO doses of 100, 200, or 400 mg/kg. PC doses in mice were 100 or 1000 mg/kg, and both species were given a 1000 mg/kg PC dose with a 50% (w/w) aqueous solution (2 ml/kg) to simulate antifreeze exposure. Results from this study have shown that orally-administered EG is very rapidly and almost completely absorbed in both rats and mice, with a bioavailable fraction of 92-100% in rats and similar percentages at the higher doses in mice. In contrast, the absorption of cutaneously applied EG is comparatively slow in both species. A species difference in the overall absorption of PC doses was demonstrated, with higher recoveries of 14C observed after PC doses in mice than for rats and a greater penetration of 14C after applying a 50% aqueous PC dose in mice than in rats, as evidenced by quantifiable plasma 14C concentrations only in mice. The major metabolites in both rats and mice are CO2 and glycolate. Oxidative metabolic pathways are saturated at high PO doses in both species, resulting in a shift from exhaled CO2 as the major excretion route to urinary excretion. The capacity to metabolize more completely EG to CO2 at low doses seems to be greater in the mouse than in the rat, as evidenced by the absence of urinary oxalate from EG-dosed female mice, and saturation of metabolic pathways at a comparatively lower dose in mice than for rats. This evidence suggests that dose-dependent changes in EG excretion in female Sprague-Dawley rats and CD-1 mice probably resulted from capacity-limited effects on EG metabolic pathways for the production of CO2 and a compensatory urine clearance of glycolate. Results from the present study corroborate previous observations in rats for the lower doses, but demonstrate a substantial difference in single-dose pharmacokinetics for IV and PO 1000 mg/kg doses in mice vs. rats. In summary, these data indicate that mice show a nonlinear plasma disposition of total radioactivity (EG and its metabolites) as dose is increased, whereas in rats plasma kinetics were linear over the dose range evaluated, whereas excretion kinetic patterns were nonlinear in both species as dose is increased.