Computation-Assisted Molecularly Imprinted Polymer Synthesis for Extraction of Naltrexone from Urine Using Experimental Design and Determination by UPLC-DAD.
To design a molecularly imprinted polymer (MIP) for naltrexone, calculations were performed using Gaussian 03 software, and the interaction energy (ΔE) of template-monomer complexes was estimated using the density functional theory method with the B3LYP function and 6-311G (d) basis set. The effect of different solvents in the polymerization process was studied using the polarizable continuum model. It was shown that five molecules of methacrylic acid gave the largest ΔE with tetrahydrofuran as the polymerization solvent. Effective factors of the removal efficiency of naltrexone by the MIP were selected using a central composite design, and thereafter, the optimization of significant factors was performed by response surface methodology. The results predicted through these models showed good agreement with experimental values. The adsorption amount, selectivity distribution coefficient, and selectivity coefficient were found to be 11.60 mg/g, 35.31, and 2.27, respectively. Experiments of naltrexone adsorption onto the MIP were in accordance with the first-order and Langmuir-Freundlich adsorption models. By applying the data to the Scatchard equation, the KD and Qmax were determined as 526.31 mg/L and 19.47 mg/g, respectively.


Structural interpretation of the 31P NMR chemical shifts in thiophosphate and phosphate: key effects due to spin-orbit and explicit solvent.
Structural interpretation of the 31P NMR shifts measured in O,O-diethyl thiophosphate (PT), 5,5-dimethyl-2-mercapto-1,3,2-dioxaphosphorinane 2-oxide (cPT), diethylphosphate (P) and 5,5-dimethyl-2-hydroxy-1,3,2-dioxaphosphinane 2-oxide (cP) was obtained by means of theoretical calculations including the effects of geometry, molecular dynamics, and solvent, relativistic effects and the effect of NMR reference. NMR calculations employed the B3LYP, BP86, BPW91, M06-2X, PBE0, MP2, and HF methods, the Iglo-n (n = II, III), cc-pVnZ (n = D, T, Q, 5), and pcS-n (n = 0, 1, 2, 3, 4) Gaussian-type basis sets and the Slater-type QZ4P atomic basis. Water solvent was described explicitly and/or implicitly. The effects due to molecular dynamics were calculated using molecular dynamics simulations with the GAFF force field and the TIP3P water molecules, and alternatively by means of the zero-point ro-vibrational averaging. Relativistic effects included the spin-orbit calculated within the two-component zero-order relativistic approximation and the effect with the four-component DFT method. Optimal geometries and large-amplitude dynamical motions within the "opened" PT and P molecules contrasted with notably different geometries and confined dynamical motions within the cPT and cP "closed" molecules. These structure-dynamical differences together with the different chemical structures of thiophosphate and phosphate due to a non-esterified sulphur or oxygen atom within the group considerably affected the magnitudes of 31P NMR shifts. The theoretical calculations enabled accurate and reliable structure-dynamical interpretation of the measured 31P NMR shifts. The effects due to explicit solvent and relativity turned out to be indispensable for obtaining accurate 31P NMR shifts particularly in the thiophosphates. Replacement of the non-esterified oxygen atom in the phosphate with sulphur makes NMR shielding of the phosphorus atom qualitatively different as compared to the NMR shielding of the phosphorus atom in phosphate, H3PO4 and PH3.


Multiple functions for Drosophila Mcm10 suggested through analysis of two Mcm10 mutant alleles.
DNA replication and the correct packaging of DNA into different states of chromatin are both essential processes in all eukaryotic cells. High-fidelity replication of DNA is essential for the transmission of genetic material to cells. Likewise the maintenance of the epigenetic chromatin states is essential to the faithful reproduction of the transcriptional state of the cell. It is becoming more apparent that these two processes are linked through interactions between DNA replication proteins and chromatin-associated proteins. In addition, more proteins are being discovered that have dual roles in both DNA replication and the maintenance of epigenetic states. We present an analysis of two Drosophila mutants in the conserved DNA replication protein Mcm10. A hypomorphic mutant demonstrates that Mcm10 has a role in heterochromatic silencing and chromosome condensation, while the analysis of a novel C-terminal truncation allele of Mcm10 suggests that an interaction with Mcm2 is not required for chromosome condensation and heterochromatic silencing but is important for DNA replication.


Total isovolumic time relates to exercise capacity in patients with transposition of the great arteries late after atrial switch procedures.
Systemic right ventricular systolic dysfunction is common late after atrial switch surgery for transposition of the great arteries. Total isovolumic time is the time that the ventricle is neither ejecting nor filling and is calculated without relying on geometric assumptions. We assessed resting total isovolumic time in this population and its relationship to exercise capacity. A total of 40 adult patients with transposition of the great arteries after atrial switch - and 10 healthy controls - underwent transthoracic echocardiography and cardiopulmonary exercise testing from January, 2006 to January, 2009. Resting total isovolumic time was measured in seconds per minute: 60 minus total ejection time plus total filling time. The mean age was 31.6 plus or minus 7.6 years, and 38.0% were men. There were 16 patients (40%) who had more than or equal to moderate systolic dysfunction of the right ventricle. Intra- and inter-observer agreement was good for total isovolumic time, which was significantly prolonged in patients compared with controls (12.0 plus or minus 3.9 seconds per minute versus 6.0 plus or minus 1.8 seconds per minute, p-value less than 0.001) and correlated significantly with peak oxygen consumption (r equals minus 0.63, p-value less than 0.001). The correlation strengthened (r equals minus 0.73, p-value less than 0.001) after excluding seven patients with exercise-induced cyanosis. No relationship was found between exercise capacity and right ventricular ejection fraction or long-axis amplitude. Resting isovolumic time is prolonged after atrial switch for patients with transposition of the great arteries. It is highly reproducible and relates well to exercise capacity.


Changes in the force-sharing pattern induced by modifications of visual feedback during force production by a set of fingers.
We investigated force-sharing among three fingers which acted in parallel and produced ramp profiles of total force from zero to the maximal voluntary force. The feedback to the subject was provided by a visual signal on the monitor and could correspond to the sum of forces of all the fingers or to the sum of forces of two fingers, while the force of the third finger was added with a coefficient 2 or 0.5. If the subjects did not know about the distorted feedback, they showed a template-sharing pattern within the whole range of total force values. This pattern did not depend on which finger force was multiplied and by which coefficient. If the subjects knew in advance how the feedback signal would be calculated, they tried to perform the task using either only the finger whose force was multiplied by 2 or two fingers when the force of the third one was multiplied by 0.5. Further into the trial, however, they were unable to track the ramp pattern using only one or two fingers and demonstrated a search activity that could continue until the end of the trial or lead eventually to a three-finger sharing pattern similar to the template pattern used in cases of undistorted feedback. We conclude that the limited number of preferred sharing pattern within the studied task reflects an organization of the fingers into a structural unit (involving one, two, or all three fingers) by the central nervous system. The availability of structural units defines the presence of stable solutions available for the system.


Information needs and prostate cancer: the development of a systematic means of identification.
To design, from first principles, a valid and reliable scale for assessing the importance of specific items of information needed by patients with prostate cancer that would be straightforward to use in clinical settings, as despite its prevalence, there is little research focusing specifically on the information needs associated with prostate cancer. Several stages of consultation and modification were used to inform the development of a scale which was then piloted on 96 patients with prostate cancer. Respondents were asked to rate the importance they placed on a range of prostate cancer-related topics of information, and the extent to which they felt these information needs had been met. The construct and content validity of the instrument were established and an exploratory factor analysis used to guide restructuring of the tool. Internal consistency/reliability was calculated using Cronbach's alpha. Using the scale showed that men with prostate cancer placed considerable importance on a broad range of information needs, most of which had been inadequately met. Age had a significant influence on the overall importance attributed to information, with younger patients having more need for information. The construct and content validity of the instrument were established. The factor analysis revealed four discrete factors which together explained > 68% of the variance, termed 'basics of prostate cancer care', 'disease management', 'physical well-being' and 'self-help'. Internal consistency/reliability was satisfactory (alpha = 0.91). The basis of a tool capable of ascertaining the information needs of patients with prostate cancer was developed; it may offer clinicians a valid means of ascertaining information preferences and hence potentially enhance the quality of service provided. Further research is now required to refine the tool and test the effect of its longitudinal use in clinical practice on patient satisfaction and outcome.


[Ultrastructure of Clostridium tetani. I. Structural features of the surface structures of vegetative cells].
Crest-like structures formed by internal layer of cell wall and cytoplasmic membrane were revealed in G1.tetani 471 by electron microscopy with the use of negative contrasting, ultrathin sections and freezing-etching. The transverse section of these crest-like structures was 56.3 nm and they were localized 4 to 6 in one row girdling the protoplast in different directions. Ring-like subunits located in rows with the periodicity of 5.9 nm, perpendicularly to the long axis of the cell, were revealed on the surface of the cell wall.


How can philosophy be a true cognitive science discipline?
Although philosophy has been only a minor contributor to cognitive science to date, this paper describes two projects in naturalistic philosophy of mind and one in naturalistic philosophy of science that have been pursued during the past 30 years and that can make theoretical and methodological contributions to cognitive science. First, stances on the mind-body problem (identity theory, functionalism, and heuristic identity theory) are relevant to cognitive science as it negotiates its relation to neuroscience and cognitive neuroscience. Second, analyses of mental representations address both their vehicles and their contents; new approaches to characterizing how representations have content are particularly relevant to understanding the relation of cognitive agents to their environments. Third, the recently formulated accounts of mechanistic explanation in philosophy of science both provide perspective on the explanatory project of cognitive science and may offer normative guidance to cognitive science (e.g., by providing perspective on how multiple disciplinary perspectives can be integrated in understanding a given mechanism).


Evaluation of the potential biological toxicities of aqueous extracts from red tide phytoplankton cultures in in vitro and in vivo systems.
The biological toxic potentials of aqueous extracts from the dinophycean flagellates Gymnodinium impudicum and Alexandrium affine and the raphidophycean flagellate Chattonella ovata were examined in both in vitro and in vivo systems. Interestingly, the extract from A. affine was the only one that showed potent cytotoxicities towards HeLa, Vero, and Neuro-2a cells in a concentration-dependent manner. Mice given intraperitoneal injections of the extracts revealed that none of the extracts exhibited serious toxicities in mice. However, temporal body weight loss was observed in the mice injected with the extract from A. affine during the early stage, and the dramatic enlargement of spleens was also observed in the mice on the 7th day after injection. Since A. affine extract showed potent hemolytic activity in vitro towards mouse erythrocytes, hemolytic anemia may be a possible mechanism responsible for the splenomegaly in the mice injected with A. affine extract. Similar marginal effects were observed in the mice injected with the extract from C. ovata; however, no significant toxic or detrimental effects were detected in the mice injected with the extract from G. impudicum. These results suggest that the extract from G. impudicum may not be contaminated with detectable levels of biologically hazardous compounds and may be relatively safe compared with the other two extracts.


Loss of rat liver microsomal cytochrome P-450 during methimazole metabolism. Role of flavin-containing monooxygenase.
The role of flavin-containing monooxygenase (FMO) in the decrease in cytochrome P-450 content during the microsomal metabolism of methimazole (N-methyl-2-mercaptoimidazole) was investigated by heat inactivation of FMO. Incubation of liver microsomes from untreated Fischer 344 rats with NADPH and methimazole resulted in a 25% loss of cytochrome P-450 detectable as its ferrous-carbon monoxide complex. The same extent of cytochrome P-450 loss was observed with 1 and 20 mM methimazole, suggesting saturation of the process. There was no significant loss of cytochrome P-450 when microsomal FMO was heat-inactivated prior to incubation with NADPH and methimazole. Heat pretreatment of the microsomes did not affect cytochrome P-450 concentrations and cytochrome P-420 was not observed. These results indicate that FMO-catalyzed metabolism of methimazole is necessary for the loss of cytochrome P-450 in microsomes from untreated rats. Sulfite and N-methylimidazole, the ultimate products of methimazole metabolism, did not cause a significant loss of cytochrome P-450. There was no loss of cytochrome P-450 when glutathione was included in the incubation with methimazole, suggesting that cytochrome P-450 loss was due to an interaction with oxygenated metabolites of methimazole formed by FMO. Losses of cytochrome P-450 were also observed after incubation of microsomes from phenobarbital- (31%) of beta-naphthoflavone-pretreated rats (44%) with NADPH and methimazole. In contrast to microsomes from untreated rats, heat inactivation of FMO did not prevent the loss of cytochrome P-450 in microsomes from the pretreated rats. These results indicate that both phenobarbital and beta-naphthoflavone induce isozymes of cytochrome P-450 capable of directly activating methimazole.


Exploring the uncertainties in cancer risk assessment using the integrated probabilistic risk assessment (IPRA) approach.
Current methods for cancer risk assessment result in single values, without any quantitative information on the uncertainties in these values. Therefore, single risk values could easily be overinterpreted. In this study, we discuss a full probabilistic cancer risk assessment approach in which all the generally recognized uncertainties in both exposure and hazard assessment are quantitatively characterized and probabilistically evaluated, resulting in a confidence interval for the final risk estimate. The methodology is applied to three example chemicals (aflatoxin, N-nitrosodimethylamine, and methyleugenol). These examples illustrate that the uncertainty in a cancer risk estimate may be huge, making single value estimates of cancer risk meaningless. Further, a risk based on linear extrapolation tends to be lower than the upper 95% confidence limit of a probabilistic risk estimate, and in that sense it is not conservative. Our conceptual analysis showed that there are two possible basic approaches for cancer risk assessment, depending on the interpretation of the dose-incidence data measured in animals. However, it remains unclear which of the two interpretations is the more adequate one, adding an additional uncertainty to the already huge confidence intervals for cancer risk estimates.


KLB dysregulation mediates disrupted muscle development in intrauterine growth restriction.
Intrauterine growth restriction (IUGR) is a leading cause of neonatal morbidity and mortality in humans and domestic animals. Developmental adaptations of skeletal muscle in IUGR lead to increased risk of premature muscle loss and metabolic disease in later life. Here, we identified β-Klotho (KLB), a fibroblast growth factor 21 (FGF21) co-receptor, as a novel regulator of muscle development in IUGR. Using the pig as a naturally-occurring disease model, we performed transcriptome-wide profiling of fetal muscle (day 90 of pregnancy) from IUGR and normal-weight (NW) littermates. We found that, alongside large-scale transcriptional changes comprising multiple developmental, tissue injury and metabolic gene pathways, KLB was increased in IUGR muscle. Moreover, FGF21 concentrations were increased in plasma in IUGR fetuses. Using cultures of fetal muscle progenitor cells (MPCs), we showed reduced myogenic capacity of IUGR compared to NW muscle in vitro, as evidenced by differences in fusion indices and myogenic transcript levels, as well as mechanistic target of rapamycin (mTOR) activity. Moreover, transfection of MPCs with KLB small interfering RNA promoted myogenesis and mTOR activation, whereas treatment with FGF21 had opposite and dose-dependent effects in porcine and also in human fetal MPCs. In conclusion, our results identify KLB as a novel and potentially critical mediator of impaired muscle development in IUGR, through conserved mechanisms in pigs and humans. Our data shed new light onto the pathogenesis of IUGR, a significant cause of lifelong ill-health in humans and animals. KEY POINTS: Intrauterine growth restriction (IUGR) is associated with large-scale transcriptional changes in developmental, tissue injury and metabolic gene pathways in fetal skeletal muscle. Levels of the fibroblast growth factor 21 (FGF21) co-receptor, β-Klotho (KLB) are increased in IUGR fetal muscle, and FGF21 concentrations are increased in IUGR fetal plasma. KLB mediates a reduction in muscle development through inhibition of mechanistic target of rapamycin signalling. These effects of KLB on muscle cells are conserved in pig and human, suggesting a vital role of this protein in the regulation of muscle development and function in mammals.


Betaine lowers elevated s-adenosylhomocysteine levels in hepatocytes from ethanol-fed rats.
Previous studies showed that chronic ethanol administration inhibits methionine synthase activity, resulting in impaired homocysteine remethylation to form methionine. This defect in homocysteine remethylation was shown to increase plasma homocysteine and to interfere with the production of hepatic S-adenosylmethionine (SAM) in ethanol-fed rats. These changes were shown to be reversed by the administration of betaine, an alternative methylating agent. This study was undertaken to determine additional effects of ethanol on methionine metabolism and their functional consequences. The influences of methionine loading and betaine supplementation were also evaluated. Adult Wistar rats were fed ethanol or a control Lieber-DeCarli liquid diet for 4 wk, and metabolites of the methionine cycle were measured in vitro in isolated hepatocytes under basal and methionine-supplemented conditions. S-Adenosylhomocysteine (SAH) concentrations were elevated in hepatocytes isolated from ethanol-fed rats compared with controls and in hepatocytes from both groups when supplemented with methionine. The addition of betaine to the methionine-supplemented incubation media reduced the elevated SAH levels. The decrease in the intracellular SAH:SAM ratio due to ethanol consumption inhibited the activity of the liver-specific SAM-dependent methyltransferase, phosphatidylethanolamine methyltransferase. Our data indicate that betaine, by remethylating homocysteine and removing SAH, overcomes the detrimental effects of ethanol consumption on methionine metabolism and may be effective in correcting methylation defects and treating liver diseases.


Personalized Risk Prediction in Clinical Oncology Research: Applications and Practical Issues Using Survival Trees and Random Forests.
A crucial component of making individualized treatment decisions is to accurately predict each patient's disease risk. In clinical oncology, disease risks are often measured through time-to-event data, such as overall survival and progression/recurrence-free survival, and are often subject to censoring. Risk prediction models based on recursive partitioning methods are becoming increasingly popular largely due to their ability to handle nonlinear relationships, higher-order interactions, and/or high-dimensional covariates. The most popular recursive partitioning methods are versions of the Classification and Regression Tree (CART) algorithm, which builds a simple interpretable tree structured model. With the aim of increasing prediction accuracy, the random forest algorithm averages multiple CART trees, creating a flexible risk prediction model. Risk prediction models used in clinical oncology commonly use both traditional demographic and tumor pathological factors as well as high-dimensional genetic markers and treatment parameters from multimodality treatments. In this article, we describe the most commonly used extensions of the CART and random forest algorithms to right-censored outcomes. We focus on how they differ from the methods for noncensored outcomes, and how the different splitting rules and methods for cost-complexity pruning impact these algorithms. We demonstrate these algorithms by analyzing a randomized Phase III clinical trial of breast cancer. We also conduct Monte Carlo simulations to compare the prediction accuracy of survival forests with more commonly used regression models under various scenarios. These simulation studies aim to evaluate how sensitive the prediction accuracy is to the underlying model specifications, the choice of tuning parameters, and the degrees of missing covariates.


[Organization of tuberculosis-controlling work and its efficiency in the penitentiaries of the Ivanovo Region].
The incidence rate of tuberculosis is high in the penitentiaries of the Ivanovo Region and in 2000 it was 32 times greater than that in the Region (3122 versus 96.7 per 100,000). Fluorography remains to be the main method of detection of pulmonary tuberculosis. It is performed on day 3 of admission of jail, then every 6 months. Fluorography allows one to identify 85-90% of the patients with pulmonary tuberculosis. In the fluorgraphic interval, the sputum from patients with symptoms of suspected tuberculosis is tested by the Ziehl-Neelsen procedure, which makes it possible to detect more than 15% of the tuberculosis patients who isolate bacteria. The patients are treated with the routine regimens including 4 first-line drugs. In 1997-2000, the rates of bacterial isolation session and decay cavity closure were in the range of 82.1-93 and 66.7-87.5%, respectively. Integration with civil tuberculosis service, the social security administration of the region, and the regional branch of the red-cross society has proved to be highly effective. The proportion of patients with tuberculosis who continued their treatment in tuberculosis facilities after release increased from 51.1% in 2000 to 80% in 2001.


[Optimization of nutrient medium for cultivation of Bacillus subtilis IMV V-7023].
Nutrient medium for cultivation of Bacillus subtilis IMV V-7023 was optimized by the method of the orthogonal Latin rectangles. Optimum concentrations were investigated in the medium of carbon source (15.0 g/l of molasses), nitrogen (2.0 g/l of corn extract) and content of phosphorus-containing inorganic salts (0.4 g/l). When growing this strain in periodic conditions at 28 degrees C, the value of oxygen mass transport in the liquid (0.4-0.6 g O2/l per 1 h) and the initial number of cells of each bacterial species was 1 x 10(6) cells/ml. The numbers of bacteria reached the maximum after 24 hours of cultivation and corresponded to 1.3 x 10(10) cells/ml. Nutrient medium that was optimized can be recommended for cultivation of B.subtilis IMV V-7023 in production conditions. The paper is presented in Russian.


Constitutive activation of the Wnt canonical pathway in mantle cell lymphoma.
Aberrations of the Wnt canonical pathway (WCP) are known to contribute to the pathogenesis of various types of cancer. We hypothesize that these defects may exist in mantle cell lymphoma (MCL). Both the upstream and downstream aspects of WCP were examined in MCL cell lines and tumors. Using WCP-specific oligonucleotide arrays, we found that MCL highly and consistently expressed Wnt3 and Wnt10. beta-catenin, a transcriptional factor that is a downstream target of WCP, is localized to the nucleus and transcriptionally active in all 3 MCL cell lines examined. By immunohistochemistry, 33 (52%) of 64 MCL tumors showed nuclear localization of beta-catenin, which significantly correlated with the expression of the phosphorylated/inactive form of GSK3beta (p-GSK3beta; P = .011, Fisher). GSK3beta inactivation is directly linked to WCP stimulation, since addition of recombinant sFRP proteins (a naturally occurring decoy for the Wnt receptors) resulted in a significant decrease in p-GSK3beta. Down-regulation of DvL-2 (an upstream signaling protein in WCP) by siRNA or selective inhibition of beta-catenin using quercetin significantly decreased cell growth in MCL cell lines. To conclude, WCP is constitutively activated in a subset of MCL and it appears to promote tumorigenesis in MCL.


The health of rectal cancer patients in the community.
Surviving rectal cancer patients living in five health districts were identified through cancer registrations. Four hundred and twenty (91%) were interviewed at home by health visitors to assess their physical, social and psychological health. In most respects, the health of rectal cancer patients overall was similar to that of the general population of the same age, sex and social circumstances. However, those with a colostomy (63%) were more likely to be severely depressed, socially isolated and stigmatized. They also suffered more from physical problems, particularly urinary symptoms, flatus and offensive bowel motions. The pressure of a colostomy was associated with reduced sexual capacity, though not with lack of sexual interest. The results confirm the high psychological and social costs of a permanent colostomy.


Arrhythmogenic right ventricular cardiomyopathy caused by deletions in plakophilin-2 and plakoglobin (Naxos disease) in families from Greece and Cyprus: genotype-phenotype relations, diagnostic features and prognosis.
To evaluate clinical disease expression, non-invasive diagnosis, and prognosis in families with dominant vs. recessive arrhythmogenic right ventricular cardiomyopathy (ARVC) due to mutations in related desmosomal proteins plakophilin-2 (PKP2) and plakoglobin (JUP), respectively. One hundred and eighty-seven individuals belonging to ARVC families, four with dominant PKP2 mutations and 12 with recessive JUP mutation underwent serial non-invasive cardiac assessment. Survival and arrhythmic events were evaluated prospectively up to 21 years (median 8.5 years). Sixteen of 22 PKP2 carriers and all 26 homozygous JUP carriers fulfilled the diagnostic criteria for ARVC, the youngest by the age of 13 years. Clinical disease expression did not differ significantly between PKP2 and JUP carriers. T-wave inversion in leads V1-V3, right ventricular wall motion abnormalities, and frequent ventricular extrasystoles were the most sensitive/specific markers for identification of mutation carriers. QRS dispersion > or =40 ms was an independent predictor of syncope but not of sudden death. Mutations in PKP2 and JUP express similar cardiac phenotype. Non-invasive family screening may largely be based on T-wave inversion, right ventricular wall motion abnormalities, and frequent ventricular extrasystoles to identify mutation carriers.


Importance of relationship quality and communication on foodservice for the elderly.
In order to promote foodservice for the elderly, foodservice managers in Continuing Care Retirement Communities (CCRCs) must identify the main factors to enhance the satisfaction and behavioral intentions with food service. The purpose of this study was to investigate the relationships between relationship quality (consisting of trust, commitment, and satisfaction) and communication in the formation of elderly's behavioral intentions with food services at CCRCs. A survey was administered to residents in two CCRCs and a total of 327 residents participated. A tested structural equation model exhibited good model fit and explanatory power of the study construct. Satisfaction directly influenced word-of-mouth and service quality has an influence on commitment. Commitment was a significant determinant of behavioral intentions to eat more often in the dining room. Also, communication showed positive association with trust. The results provided strong evidence for the importance of satisfaction and communication as a consequence of relationship marketing efforts. Suggestions for future research to better understand the elderly' behavioral intention judgments were given.


Signal amplification and transduction in phytochrome photosensors.
Sensory proteins must relay structural signals from the sensory site over large distances to regulatory output domains. Phytochromes are a major family of red-light-sensing kinases that control diverse cellular functions in plants, bacteria and fungi. Bacterial phytochromes consist of a photosensory core and a carboxy-terminal regulatory domain. Structures of photosensory cores are reported in the resting state and conformational responses to light activation have been proposed in the vicinity of the chromophore. However, the structure of the signalling state and the mechanism of downstream signal relay through the photosensory core remain elusive. Here we report crystal and solution structures of the resting and activated states of the photosensory core of the bacteriophytochrome from Deinococcus radiodurans. The structures show an open and closed form of the dimeric protein for the activated and resting states, respectively. This nanometre-scale rearrangement is controlled by refolding of an evolutionarily conserved 'tongue', which is in contact with the chromophore. The findings reveal an unusual mechanism in which atomic-scale conformational changes around the chromophore are first amplified into an ångstrom-scale distance change in the tongue, and further grow into a nanometre-scale conformational signal. The structural mechanism is a blueprint for understanding how phytochromes connect to the cellular signalling network.


Isolation and properties of a puromycin acetyltransferase from puromycin-producing Streptomyces alboniger.
Puromycin 2"-N-acetyltransferase was isolated from cell extracts of puromycin-producing Streptomyces alboniger KCC S-0309 by ammonium sulfate fractionation, heat treatment to eliminate contaminant proteins and chromatography on DEAE-Toyopearl 650S. After PAGE (polyacrylamide gel electrophoresis) of the final fraction, a single protein band corresponding to puromycin 2"-N-acetyltransferase was detected. The molecular weight of the enzyme determined by SDS-PAGE and Sephadex G-150 chromatography was about 21,000 and 85,000, respectively, suggesting that the enzyme consisted of four subunits. The isoelectric point and the optimum pH for reaction were 6.2 and 7.7, respectively. The Km values for puromycin and acetyl coenzyme A were 40 microM and 67 microM, respectively. The enzyme was thermostable up to 70 degrees C for 12 minutes. It was shown, by using an in vitro protein synthesizing system from a puromycin-susceptible organism S. flavotricini subsp. pseudochromogenes V-13-1, that the isolated puromycin 2"-N-acetyltransferase could protect polyphenylalanine synthesis from inhibition by puromycin.


Energy-level alignment at organic heterointerfaces.
Today's champion organic (opto-)electronic devices comprise an ever-increasing number of different organic-semiconductor layers. The functionality of these complex heterostructures largely derives from the relative alignment of the frontier molecular-orbital energies in each layer with respect to those in all others. Despite the technological relevance of the energy-level alignment at organic heterointerfaces, and despite continued scientific interest, a reliable model that can quantitatively predict the full range of phenomena observed at such interfaces is notably absent. We identify the limitations of previous attempts to formulate such a model and highlight inconsistencies in the interpretation of the experimental data they were based on. We then develop a theoretical framework, which we demonstrate to accurately reproduce experiment. Applying this theory, a comprehensive overview of all possible energy-level alignment scenarios that can be encountered at organic heterojunctions is finally given. These results will help focus future efforts on developing functional organic interfaces for superior device performance.


Beta-thalassaemia in south-western Spain: high frequency of G-->A (IVS I-1) mutation.
Huelva province is situated in southwestern Spain; its historic and geographic characteristics contribute to the existence of erythrocyte genetic anomalies, such as beta thalassaemia. We have carried out a prenatal study of microcytosis, with a preliminary beta-gene analysis. Our findings show a beta-thalassaemia trait prevalence of 0.81%. In the molecular research there was a high frequency of G-->A (IVS I-1) injury: 55%. By comparing the rates of this mutation neighbouring Cáceres province in western Spain (47%) and Algarve in south Portugal (32%), it demonstrates that this beta-gene defect has a high frequency in the southwest of the Iberian Peninsula.


Cytoskeleton and mechanotransduction in the pathophysiology of ventilator-induced lung injury.
Although mechanical ventilation is an important therapy, it can result in complications. One major complication is ventilator-induced lung injury, which is caused by alveolar hyperdistension, leading to an inflammatory process, with neutrophilic infiltration, hyaline membrane formation, fibrogenesis and impaired gas exchange. In this process, cellular mechanotransduction of the overstretching stimulus is mediated by means of the cytoskeleton and its cell-cell and cell-extracellular matrix interactions, in such a way that the mechanical stimulus of ventilation is translated into an intracellular biochemical signal, inducing endothelial activation, pulmonary vascular permeability, leukocyte chemotaxis, cytokine production and, possibly, distal organ failure. Clinical studies have shown the relationship between pulmonary distension and mortality in patients with ventilator-induced lung injury. However, although the cytoskeleton plays a fundamental role in the pathogenesis of ventilator-induced lung injury, there have been few in vivo studies of alterations in the cytoskeleton and in cytoskeleton-associated proteins during this pathological process.


[Aging and health in Sub-Saharan Africa: an urgent agenda for international cooperation].
The Sub-Saharan part of the African continent is the area that has the highest disease burden in the world and is the only region of the planet where it is expected that the number of poor people will increase in the coming decades. The countries of this region, to different degrees, experience slow process of population aging but at the same time, it is the are where the elderly population grows fastest in absolute numbers. Based on a review of the literature, an attempt was made to highlight the social and demographic situation in which the elderly live in the Sub-Saharan region and the main challenges faced by local governments to overcome the complex problems affecting society as a whole. It was found that public policies geared to this segment of the population in the region do not represent a priority and, consequently, are unlikely to be included in the current agenda of international cooperation.


Types of mucosal metaplasia in relation to the histogenesis of gastric carcinoma.
One hundred thirty-nine surgically resected stomachs and 16 stomachs obtained at necropsy in China were studied histochemically to evaluate the types and distribution of mucosal metaplasia in relation to benign and malignant disease. Three classes of mucin were distinguished: small-intestinal (N-acylated sialomucin); colonic (sulfated and O-acylated sialomucin); and gastric (neutral mucin). Small-intestinal type of metaplasia (SIM) occurred in 95% of instances of atrophic gastritis, 36% of cases of superficial gastritis, 60% of cases of gastric ulcers, and 39% of cases of duodenal ulcers. Colonic type of metaplasia (CIM) was less common and was found in only 11% of benign lesions. It was located in the antrum, particularly on the lesser curvature. Mucin was present in 66 of 84 carcinomas. The stomach tumors fell into three histochemical groups: colonic (acid mucin), 32; gastric (neutral mucin), 19; and mixed (acidic and neutral), 15. In 94% (30/32) of colonic type of gastric carcinoma, CIM was present. Thus, CIM is related to development of colonic type of gastric carcinoma, whereas SIM is a nonspecific reaction to mucosal damage.


Pseudo-equilibrium equations for calcium phosphate precipitation with multi-unit particles.
The chemistry underlying bone mineral formation in vertebrates is the reaction of calcium phosphate precipitation. In a near-neutral solution, an amorphous phase and hydroxyapatite nanoparticles appear successively, and the reaction system containing either of the two kinds of precipitates is in a non-equilibrium state. Here, we propose a pseudo-equilibrium approach to the solution chemistry of the precipitation reactions. We employed two series of reaction systems, collected samples at various stages, and analyzed the solution chemistry data on the basis of a simplified model of reaction. We derived two types of pseudo-equilibrium equations from the two series, respectively. These equations reveal the existence of multiple structural units in a precipitate particle and correlate the ionic product with the surface proportion per structural unit (m). The surface proportion, in turn, is related to the whole particle through a particle-surface equation. Notably, the two types of pseudo-equilibrium constants have the common expression of "Kd = ionic product" if the number of the structural units (u) is large enough. Together, these findings have revealed some aspects of the non-equilibrium thermodynamics of precipitation reactions, indicating the solution chemistry route to the equilibrium state. The concept of the multi-unit particle may shed new light on the study of precipitation reactions of other slightly soluble electrolytes. And the relationship between the ionic product and the surface proportion of a structural unit is not only fundamental in chemistry, but may also apply to non-equilibrium systems in nature and biology, such as marine sedimentation, human vascular calcification, and bone mineral metabolism.


Bioactivity of a radical scavenger bis(pyrazolium p-toluenesulphonate) on ctDNA and certain microbes: a combined experimental and theoretical analysis.
A small organic molecule, bis(pyrazolium p-toluenesulphonate) (BPPTS), was crystallized, characterized and used to scavenge free radicals in biological systems. SXRD and spectroscopic analyses were used to confirm the structure of BPPTS. Methanolic and ethanolic solutions of BPPTS were used to assess the stability of the proposed drug using the UV-vis spectrophotometric technique. Optimization of the molecular structure was carried out by DFT with B3LYP/6-311++G(d,p) level of basis set. MEP and Fukui functions that elaborate theoretically the predominant electrophilic, nucleophilic and radical sites in BPPTS were correlated with experimental biological screening. BPPTS exhibits strong activity against Bacillus subtilis and Escherichia coli, comparable with all other analyzed pathogens. The free radical scavenging activity of BPPTS was assessed by both experimental studies and theoretical calculations. The binding sites of DPPH, which can bind to BPPTS, were also predicted by Fukui functions. DNA binding of BPPTS in UV-vis studies revealed the groove mode of binding due to the occurrence of hyperchromism. The phenomenon of hyperchromism was established by the Hirshfeld surface analysis of BPPTS, which confirmed the presence of π···π interactions (2.4%). Molecular docking established a positive correlation between experimental bio-screening reports and simulated data. ADMET properties were also calculated.


New analysis reexamines the value of cancer care in the United States compared to Western Europe.
Despite sharp increases in spending on cancer treatment since 1970 in the United States compared to Western Europe, US cancer mortality rates have decreased only modestly. This has raised questions about the additional value of US cancer care derived from this additional spending. We calculated the number of US cancer deaths averted, compared to the situation in Western Europe, between 1982 and 2010 for twelve cancer types. We also assessed the value of US cancer care, compared to that in Western Europe, by estimating the ratio of additional spending on cancer to the number of quality-adjusted life-years saved. Compared to Western Europe, for three of the four costliest US cancers-breast, colorectal, and prostate-there were approximately 67,000, 265,000, and 60,000 averted US deaths, respectively, and for lung cancer there were roughly 1,120,000 excess deaths in the study period. The ratio of incremental cost to quality-adjusted life-years saved equaled $402,000 for breast cancer, $110,000 for colorectal cancer, and $1,979,000 for prostate cancer-amounts that exceed most accepted thresholds for cost-effective medical care. The United States lost quality-adjusted life-years despite additional spending for lung cancer: -$19,000 per quality-adjusted life-year saved. Our results suggest that cancer care in the United States may provide less value than corresponding cancer care in Western Europe for many leading cancers.


Induction of indoleamine 2, 3-dioxygenase in human dendritic cells by a cholera toxin B subunit-proinsulin vaccine.
Dendritic cells (DC) interact with naïve T cells to regulate the delicate balance between immunity and tolerance required to maintain immunological homeostasis. In this study, immature human dendritic cells (iDC) were inoculated with a chimeric fusion protein vaccine containing the pancreatic β-cell auto-antigen proinsulin linked to a mucosal adjuvant the cholera toxin B subunit (CTB-INS). Proteomic analysis of vaccine inoculated DCs revealed strong up-regulation of the tryptophan catabolic enzyme indoleamine 2, 3-dioxygenase (IDO1). Increased biosynthesis of the immunosuppressive enzyme was detected in DCs inoculated with the CTB-INS fusion protein but not in DCs inoculated with proinsulin, CTB, or an unlinked combination of the two proteins. Immunoblot and PCR analyses of vaccine treated DCs detected IDO1mRNA by 3 hours and IDO1 protein synthesis by 6 hours after vaccine inoculation. Determination of IDO1 activity in vaccinated DCs by measurement of tryptophan degradation products (kynurenines) showed increased tryptophan cleavage into N-formyl kynurenine. Vaccination did not interfere with monocytes differentiation into DC, suggesting the vaccine can function safely in the human immune system. Treatment of vaccinated DCs with pharmacological NF-κB inhibitors ACHP or DHMEQ significantly inhibited IDO1 biosynthesis, suggesting a role for NF-κB signaling in vaccine up-regulation of dendritic cell IDO1. Heat map analysis of the proteomic data revealed an overall down-regulation of vaccinated DC functions, suggesting vaccine suppression of DC maturation. Together, our experimental data indicate that CTB-INS vaccine induction of IDO1 biosynthesis in human DCs may result in the inhibition of DC maturation generating a durable state of immunological tolerance. Understanding how CTB-INS modulates IDO1 activity in human DCs will facilitate vaccine efficacy and safety, moving this immunosuppressive strategy closer to clinical applications for prevention of type 1 diabetes autoimmunity.


Double outlet left ventricle: diagnosis with magnetic resonance imaging.
A complex congenital cardiac malformation in a female patient was evaluated several times by angiocardiography and echocardiography in childhood but a definite diagnosis was not established. Segmental analysis of the heart and the great vessels by magnetic resonance imaging when the patient was 34, however, showed a double outlet left ventricle in which the aorta was situated anterior to and to the left of the pulmonary trunk and an associated subaortic ventricular septal defect with pulmonary valve stenosis. This is the first time that this extremely rare cardiac malformation has been diagnosed by magnetic resonance imaging.


Involvement of chemokine receptors in breast cancer metastasis.
Breast cancer is characterized by a distinct metastatic pattern involving the regional lymph nodes, bone marrow, lung and liver. Tumour cell migration and metastasis share many similarities with leukocyte trafficking, which is critically regulated by chemokines and their receptors. Here we report that the chemokine receptors CXCR4 and CCR7 are highly expressed in human breast cancer cells, malignant breast tumours and metastases. Their respective ligands CXCL12/SDF-1alpha and CCL21/6Ckine exhibit peak levels of expression in organs representing the first destinations of breast cancer metastasis. In breast cancer cells, signalling through CXCR4 or CCR7 mediates actin polymerization and pseudopodia formation, and subsequently induces chemotactic and invasive responses. In vivo, neutralizing the interactions of CXCL12/CXCR4 significantly impairs metastasis of breast cancer cells to regional lymph nodes and lung. Malignant melanoma, which has a similar metastatic pattern as breast cancer but also a high incidence of skin metastases, shows high expression levels of CCR10 in addition to CXCR4 and CCR7. Our findings indicate that chemokines and their receptors have a critical role in determining the metastatic destination of tumour cells.


Lactobacillus plantarum L9 but not Lactobacillus acidophilus LA reduces tumour necrosis factor induced bacterial translocation in Caco-2 cells.
Translocation of bacteria across the intestinal barrier is important in the pathogenesis of systemic sepsis and multiple organ dysfunction syndromes. Inflammatory cytokines increase paracellular permeability that allows increased luminal bacteria to translocate across mucosal epithelium and further deteriorate the gut barrier. In order to reduce this risk, the prophylactic use of probiotics has been recently addressed. In this paper, we investigate the protective role toward tumour necrosis factor (TNF)-α induced non-pathogenic Escherichia coli translocation across Caco-2 monolayers of Lactobacillus strains. According to our experimental data, Lactobacillus plantarum L9 and Lactobacillus acidophilus LA have good capacities to adhere to Caco-2 cells. Addition of L. plantarum L9 and L. acidophilus LA to the enterocyte monolayer surface result in significant inhibition of E. coli adhesion and cell internalisation. However, L. plantarum L9 and L. acidophilus LA did not inhibit the growth of the non-pathogenic E. coli B5 after 24 h incubation. Exposure to TNF-α for 6 h caused a dramatic increase in E. coli B5 translocation across Caco-2 cells, which was uncoupled from increases in paracellular permeability. Pretreatment with L. plantarum L9 prevent TNF-α induced transcellular bacterial translocation and IL-8 production in Caco-2 cells. L. plantarum L9 also did not affect the integrity of the monolayers, as indicated by lactate dehydrogenase release, horseradish peroxidase permeability, and transepithelial electrical resistance. L. plantarum L9 showed the potential to protect enterocytes from an acute inflammatory response and therefore could be good potential prophylactic agents in counteracting bacterial translocation.


Molecular biological characterization of a highly leukaemogenic virus isolated from the mouse. III. Identity with mouse mammary tumour virus.
A highly leukaemogenic virus isolate (DMBA-LV) endogenous to the CFW/D mouse has been found to contain two viral genomes. One was closely related to the type B milk-borne mouse mammary tumour virus (MMTV) and present in tenfold excess over a type C viral genome which was only partially related to xenotropic and polytropic isolates from the CFW/D mouse as well as to the ecotropic Moloney murine leukaemia virus isolate. The thymic lymphoma cell line that produced DMBA-LV expressed high levels of MMTV viral RNA (35S and the 24S envelope mRNA). Both the virus and the virus-producing cell line expressed multiple species of type C viral RNA. Similar species of type C viral RNA were also associated with non-infectious, non-leukaemogenic viral particles present in both normal lymphoid cells and in a MMTV-free thymic lymphoma cell line established from a second chemical carcinogen-induced tumour.


Protonation and anion-binding properties of aromatic sulfonylurea derivatives.
In this work the anion-binding properties of three aromatic sulfonylurea derivatives in acetonitrile and dimethyl sulfoxide were explored by means of NMR titrations. It was found that the studied receptors effectively bind anions of low basicity (Cl-, Br-, I-, NO3 - and HSO4 -). The stoichiometry of the complexes with receptors containing one binding site was 1 : 1 exclusively, whereas in the case of the receptor containing two sulfonylurea groups 1 : 2 (receptor : anion) complexes were also detected in some cases. The presence of strongly basic anions (acetate and dihydrogen phosphate) led to the deprotonation of the sulfonylurea moiety. This completely hindered its anion-binding properties in DMSO and only proton transfer occurred upon the addition of basic anions to the studied receptors. In MeCN, a complex system of equilibria including both ligand deprotonation and anion binding was established. Since ionisation of receptors was proven to be a decisive factor defining the behaviour of the sulfonylurea receptors, their pK a values were determined using several deprotonation agents in both solvents. The results were interpreted in the context of receptor structures and solvent properties and applied for the identification of the interactions with basic anions.


An adult system versus a Bain system: comparative ability to deliver minute ventilation to an infant lung model with pressure-limited ventilation.
We compared the efficacy of an adult circle system versus a Bain system to deliver minute ventilation (V(E)) to an infant test lung model using pressure-limited ventilation. To simulate a wide variety of potential infant clinical states, V(E) was measured with two compliances: at peak inspiratory pressures (PIP) of 20, 30, 40, and 50 cm H2O and at respiratory rates (RR) of 20, 30, 40, and 50 breaths/min. Each measurement was made three times, and their average was used for analysis. Data were analyzed using the multiple regression technique. In both normal and low-compliance lung models, V(E) was nearly identical between adult circle and Bain systems (P = 0.67 for normal compliance model, P = 0.89 for low-compliance model). V(E) positively correlated with RR (P < 0.001), PIP (P < 0.001), and lung compliance (P < 0.001). Very high PIP or RR were required to deliver V(E) to the low-compliance lung model. The adult circle system is equivalent to the Bain system in its ability to ventilate an infant test lung over a wide range of RR, PIP, and two compliances during pressure-limited ventilation. V(E) is dependent of PIP, RR, and lung compliance. With low-compliance lungs, both systems require a high PIP. We conclude that both anesthetic systems deliver ventilation over a wide range of respiratory variables during pressure-limited ventilation in infants. We obtained results from this infant test lung study that indicate that either an adult circle breathing system or the Bain system can reliably deliver ventilation over a wide range of respiratory variables during pressure-limited ventilation in infants.


Promoting treatment for hepatitis C in people who inject drugs: A review of the barriers and opportunities.
People who inject drugs (PWIDs) comprise a significant amount of the population who are also positive for hepatitis C virus (HCV) around the world. Even though there is evidence that patients who currently inject drugs do not display altered treatment adherence or medication effectiveness, health care providers are still hesitant to treat this patient population based on perceived threats and barriers. This literature review informs of the perceived barriers associated with PWID in receiving HCV treatment and supports recommendations to address these barriers. For this review, eight scholarly articles rated levels I A through II B using John Hopkins Evidence-Based Practicing ratings consisting of randomized controlled trials, systematic reviews, and meta-analyses were selected. Both qualitative and quantitative data contributed to identifying perceived barriers and suggested course of action that should be taken to increase HCV treatment uptake among PWID. The three main perceived barriers are evidence of barriers to treatment in the PWID populations and include increased risk of reinfection, low adherence to treatment, and decreased response to treatment. Removal of these barriers by increasing education about HCV disease and treatment options to both patients and health care workers and changing current policy in health care settings to provide enhanced access to HCV treatment for PWID can promote an opportunity for successful treatment of these patients. There is a significant need for HCV treatment among PWID. Opportunities for successful treatment exist and should be adopted.


Microarray analysis indicates an important role for FABP5 and putative novel FABPs on a Western-type diet.
Liver parenchymal cells play a dominant role in hepatic metabolism and thereby total body cholesterol homeostasis. To gain insight into the specific pathways and genes involved in the response of liver parenchymal cells to increased dietary lipid levels under atherogenic conditions, changes in parenchymal cell gene expression upon feeding a Western-type diet for 0, 2, 4, and 6 weeks were determined using microarray analysis in LDL receptor-deficient mice, an established atherosclerotic animal model. Using ABI Mouse Genome Survey Arrays, we were able to detect 7,507 genes (28% of the total number on an array) that were expressed in parenchymal cells isolated from livers of LDL receptor-deficient mice at every time point investigated. Time-dependent gene expression profiling identified fatty acid binding protein 5 (FABP5) and four novel FABP5-like transcripts located on chromosomes 2, 8, and 18 as important proteins in the primary response of liver parenchymal cells to Western-type diet feeding, because their expression was 16- to 22-fold increased within the first 2 weeks on the Western-type diet. The rapid substantial increase in gene expression suggests that these FABPs may play an important role in the primary protection against the cellular toxicity of cholesterol, free fatty acids, and/or lipid oxidants. Furthermore, as a secondary response to the Western-type diet, liver parenchymal cells of LDL receptor-deficient mice stimulated glycolysis and lipogenesis pathways, resulting in a steady, more atherogenic serum lipoprotein profile (increased VLDL/LDL).


The majority of intestinal IgA+ and IgG+ plasmablasts in the human gut are antigen-specific.
Mucosal antibody responses play a major role in mediating homeostasis with the intestinal flora. It has been suggested that imbalance in the IgA+ and IgG+ intestinal B cell repertoire may be associated with the development of diseases such as inflammatory bowel disease. Despite this, little is known about the antibody specificity of human intestinal plasmablasts. Here, we have determined the reactivity profile of single isolated IgA+ and IgG+ plasmablasts from human terminal ileum using antibody cloning and in vitro expression. We found that approximately 25% of intestinal IgA and IgG plasmablast antibodies were polyreactive; the majority were antigen-specific. Antigen specificity was not only directed against enteropathogenic microbes but also against commensal microbes and self antigens. Regardless of their reactivity, all intestinal antibodies were somatically mutated and showed signs of antigen-mediated selection, suggesting that they developed from antigen-specific B cell responses. Together, our data indicate that antigen-specific immune responses to intestinal microbes are largely responsible for the maintenance of intestinal homeostasis and thus provide a basis for understanding the deregulated immune responses observed in patients with inflammatory bowel disease.


Aqueous humor fibrinolytic activity in dogs with cataracts.
To investigate fibrinolytic activity in aqueous humor (AH) of healthy and sick dogs, with and without cataracts. Prospective observational clinical study. A total of 45 dogs were included in the study. Physical and ophthalmic examinations, complete blood cell count (CBC) and serum biochemistry panel were performed in all animals. According to the ocular and systemic diagnoses, animals were classified into three groups: sick dogs without cataracts (20 dogs; 40 eyes), diabetic dogs with cataracts (11 dogs; 22 eyes), and healthy dogs with cataracts (14 dogs; 25 eyes). Bilateral AH and blood samples were collected during intraocular surgery (25 dogs; 47 eyes), or immediately after euthanasia (20 dogs; 40 eyes). Citrated samples were centrifuged and stored at -81 °C until analysis. Plasma and AH D-dimer concentration were determined using a quantitative immunoturbidimetric latex agglutination assay. A total of 108 canine samples (45 plasma and 87 AH samples) were obtained. D-dimer concentration in log-scale, in AH of eyes with diabetic cataract was significantly higher than AH of eyes with nondiabetic cataract, with a difference of 0.9 ng/mL 95% confidence interval (CI) (0.2; 1.6) P = 0.0116 and higher than that of sick animals with healthy eyes, with a estimated difference of -0.89 ng/mL 95% CI (-1.52; -0.25) P = 0.0061. Plasma D-dimer concentration was significantly higher in the group of animals with systemic disease [median 606 ng/mL, Interquartil Range (IQR) 145-1956 ng/mL] than in healthy dogs (median 47.5 ng/mL, IQR 4-250 ng/mL) (P = 0.002) and diabetic dogs (median 60.5 ng/mL, IQR 0-147.5 ng/mL) (P < 0.001). AH fibrinolysis is present in dogs, being significantly higher in animals with diabetic cataracts than in those without cataracts, and those with nondiabetic cataracts.


Serine/arginine protein-specific kinase 2 promotes leukemia cell proliferation by phosphorylating acinus and regulating cyclin A1.
Serine/arginine (SR) protein-specific kinase (SRPK), a family of cell cycle-regulated protein kinases, phosphorylate SR domain-containing proteins in nuclear speckles and mediate the pre-mRNA splicing. However, the physiologic roles of this event in cell cycle are incompletely understood. Here, we show that SRPK2 binds and phosphorylates acinus, an SR protein essential for RNA splicing, and redistributes it from the nuclear speckles to the nucleoplasm, resulting in cyclin A1 but not A2 up-regulation. Acinus S422D, an SRPK2 phosphorylation mimetic, enhances cyclin A1 transcription, whereas acinus S422A, an unphosphorylatable mutant, blocks the stimulatory effect of SRPK2. Ablation of acinus or SRPK2 abrogates cyclin A1 expression in leukemia cells and arrest cells at G(1) phase. Overexpression of acinus or SRPK2 increases leukemia cell proliferation. Furthermore, both SRPK2 and acinus are overexpressed in some human acute myelogenous leukemia patients and correlate with elevated cyclin A1 expression levels, fitting with the oncogenic activity of cyclin A1 in leukemia. Thus, our findings establish a molecular mechanism by which SR splicing machinery regulates cell cycle and contributes to leukemia tumorigenesis.


Persistent regional and racial disparities in nondirected living kidney donation.
Nondirected living donors (NDLDs) are an important and growing source of kidneys to help reduce the organ shortage. In its infancy, NDLD transplantation was clustered at a few transplant centers and rarely benefited African American (AA) recipients. However, NDLDs have increased 9.4-fold since 2000, and now are often used to initiate kidney paired donation chains. Therefore, we hypothesized that the initial geographic clustering and racial disparities may have improved. We used Scientific Registry of Transplant Recipients data to compare NDLDs and their recipients between 2008-2015 and 2000-2007. We found that NDLD increased an average of 12% per year, from 20 in 2000 to 188 in 2015 (IRR: 1.12, 95% CI: 1.11-1.13, P < .001). In 2000-2007, 18.3% of recipients of NDLD kidneys were AA; this decreased in 2008-2015 to 15.7%. NDLD transplants initially became more evenly distributed across centers (Gini 0.91 in 2000 to Gini 0.69 in 2011), but then became more clustered at fewer transplant centers (Gini 0.75 in 2015). Despite the increased number of NDLDs, racial disparities have worsened and the center-level distribution of NDLD transplants has narrowed in recent years.


Kinetics-Controlled Amphiphile Self-Assembly Processes.
Amphiphile self-assembly is an essential bottom-up approach of fabricating advanced functional materials. Self-assembled materials with desired structures are often obtained through thermodynamic control. Here, we demonstrate that the selection of kinetic pathways can lead to drastically different self-assembled structures, underlining the significance of kinetic control in self-assembly. By constructing kinetic network models from large-scale molecular dynamics simulations, we show that two largely similar amphiphiles, 1-[11-oxo-11-(pyren-1-ylmethoxy)-undecyl]pyridinium bromide (PYR) and 1-(11-((5a1,8a-dihydropyren-1-yl)methylamino)-11-oxoundecyl)pyridinium bromide (PYN), prefer distinct kinetic assembly pathways. While PYR prefers an incremental growth mechanism and forms a nanotube, PYN favors a hopping growth pathway leading to a vesicle. Such preference was found to originate from the subtle difference in the distributions of hydrophobic and hydrophilic groups in their chemical structures, which leads to different rates of the adhesion process among the aggregating micelles. Our results are in good agreement with experimental results, and accentuate the role of kinetics in the rational design of amphiphile self-assembly.


α-Expansin EXPA4 Positively Regulates Abiotic Stress Tolerance but Negatively Regulates Pathogen Resistance in Nicotiana tabacum.
Since they function as cell wall-loosening proteins, expansins can affect plant growth, developmental processes and environmental stress responses. Our previous study demonstrated that changes in Nicotiana tabacum α-expansin 4 (EXPA4) expression affect the sensitivity of tobacco to Tobacco mosaic virus [recombinant TMV encoding green fluorescent protein (TMV-GFP)] infection by Agrobacterium-mediated transient expression. In this study, to characterize the function of tobacco EXPA4 further, EXPA4 RNA interfernce (RNAi) mutants and overexpression lines were generated and assayed for their tolerance to abiotic stress and resistance to pathogens. First, the differential phenotypes and histomorphology of transgenic plants with altered EXPA4 expression indicated that EXPA4 is essential for normal tobacco growth and development. By utilizing tobacco EXPA4 mutants with abiotic stress, it was demonstrated that RNAi mutants have increased hypersensitivity to salt and drought stress. In contrast, the overexpression of EXPA4 in tobacco conferred greater tolerance to salt and drought stress, as indicated by less cell damage, higher fresh weight, higher soluble sugar and proline accumulation, and higher expression levels of several stress-responsive genes. In addition, the overexpression lines were more susceptible to the viral pathogen TMV-GFP when compared with the wild type or RNAi mutants. The induction of the antioxidant system, several defense-associated phytohormones and gene expression was down-regulated in overexpression lines but up-regulated in RNAi mutants when compared with the wild type following TMV-GFP infection. In addition, EXPA4 overexpression also accelerated the disease development of Pseudomonas syringae DC3000 on tobacco. Taken together, these results suggested that EXPA4 appears to be important in tobacco growth and responses to abiotic and biotic stress.


Bioseguridad in Mexico: Pursuing Security between Local and Global Biologies.
In the aftermath of the 2009 outbreak of H1N1 influenza, scientists in Mexico sought to develop bioseguridad, that is, to protect biological life in Mexico by safely conducting research on infectious disease. Drawing on ethnographic research in laboratories and with scientists in Mexico, I look at how scientists make claims about local differences in regulations, infrastructure, bodies, and culture. The scientists working with infectious microbes sought to establish how different microbial ecologies, human immune systems, and political and regulatory systems made the risks of research different in Mexico from other countries. In developing bioseguridad, the idea of globalized biology that animates many public health projects was undermined as scientists attended to the elements of place that affected human health and safety. Scientists argued for the importance of local biologies, generating tension with global public health projects and regulations premised on the universality of biology.


[The participation of humoral factors released by the endothelium in the development of reactive hyperemia].
In anesthetized dogs and on test-preparations of isolated vascular rings of femoral artery used for detection of appearance of vasoactive substances in venous blood, the reactive hyperemia was accompanied by the appearance of vasodilation substances in blood, the concentration of which increased with the occlusion duration. Chemical injury of endothelium by saponin, inhibition of lipoxygenase by quercetin and guanylate cyclase by methylene blue decreased the reactive hyperemia and the relaxation of test-preparation. Cyclooxygenase inhibition by indomethacin did not affect the reactive hyperemia or release of vasoactive substances. The endothelium seems to play an important role in development of reactive hyperemia induced by endothelium-derived relaxing factor release.


Structure and function of a family 10 beta-xylanase chimera of Streptomyces olivaceoviridis E-86 FXYN and Cellulomonas fimi Cex.
The catalytic domain of xylanases belonging to glycoside hydrolase family 10 (GH10) can be divided into 22 modules (M1 to M22; Sato, Y., Niimura, Y., Yura, K., and Go, M. (1999) Gene (Amst.) 238, 93-101). Inspection of the crystal structure of a GH10 xylanase from Streptomyces olivaceoviridis E-86 (SoXyn10A) revealed that the catalytic domain of GH10 xylanases can be dissected into two parts, an N-terminal larger region and C-terminal smaller region, by the substrate binding cleft, corresponding to the module border between M14 and M15. It has been suggested that the topology of the substrate binding clefts of GH10 xylanases are not conserved (Charnock, S. J., Spurway, T. D., Xie, H., Beylot, M. H., Virden, R., Warren, R. A. J., Hazlewood, G. P., and Gilbert, H. J. (1998) J. Biol. Chem. 273, 32187-32199). To facilitate a greater understanding of the structure-function relationship of the substrate binding cleft of GH10 xylanases, a chimeric xylanase between SoXyn10A and Xyn10A from Cellulomonas fimi (CfXyn10A) was constructed, and the topology of the hybrid substrate binding cleft established. At the three-dimensional level, SoXyn10A and CfXyn10A appear to possess 5 subsites, with the amino acid residues comprising subsites -3 to +1 being well conserved, although the +2 subsites are quite different. Biochemical analyses of the chimeric enzyme along with SoXyn10A and CfXyn10A indicated that differences in the structure of subsite +2 influence bond cleavage frequencies and the catalytic efficiency of xylooligosaccharide hydrolysis. The hybrid enzyme constructed in this study displays fascinating biochemistry, with an interesting combination of properties from the parent enzymes, resulting in a low production of xylose.


Atomic force microscope observation of athletes' hemoglobin imaging.
Hemoglobin (Hb) is a protein and its functional form has a tetrameric structure. This structure is the result of a combination of four sub-units called globin and indicates the dynamic interaction between them. Each subunit has a ring-shaped organic molecule called a heme that contains an iron atom; Heme is a group that mediates the reversible binding of oxygen by hemoglobin. This research was performed to observe the image of Hb by an atomic force microscope (AFM) and measure the physical function of athletes. For this purpose, based on the principle of AFM imaging, the hemoglobin crosslinking method was used to measure the morphology and size of cross-linked Hb, glutaraldehyde and Hb diameter were detected to prepare cross-linked Hb samples with different molar ratio, the activity of peroxidase was detected by Trinder reaction. The AFM was used to detect the influence of physiological environment changes such as pH, temperature, oxygen partial pressure and osmotic pressure on the absorption spectrum of Hb imaging. Results showed that the size of the uncrosslinked Hb was 6.64 nm. With the increase of the molar ratio of glutaraldehyde to Hb, the number of Hb molecules involved in the crosslinking increased, and the molecular size increased. During the crosslinking process, the aggregation of the cross-linked molecules would make the particle size of some Hb molecules reach 80-100 nm. The peak height, peak position and peak shape of the characteristic absorption peaks of pH to hemoglobin at 550 and 589 nm occurred. When the temperature changes continuously in the range of 30-55℃, the peak height of Hb absorption spectrum of normal red blood cells at 550 nm and 589 nm decreases gradually with the increase of temperature, and the peak shape at 610 nm changes obviously at 42℃, which indicates that the molecular structure of Hb changes; the absorption spectrum curve of deoxygenation disappears at 500 nm, the oxygen-binding capacity of Hb is very low, and the oxygen affinity and oxygenated hemoglobin are low (The concentration of HbO2) decreased, the osmotic pressure increased, the RBC dehydrated, the volume decreased, and the concentration of Hb increased. Conclusion: It is more accurate and comprehensive to use AFM to observe athletes' hemoglobin.


Outcomes of Esophageal Replacement: Gastric Pull-Up and Colonic Interposition Procedures.
No consensus exists about the optimal surgical technique for esophageal replacement. This study reports the surgical outcomes for the gastric pull-up and the colonic interposition procedures. A retrospective review of children undergoing esophageal replacement surgery between January 2001 and June 2015 across four different pediatric surgery centers was conducted. Data collected included indications, epidemiology, surgical technique, complications, and outcomes. Patients were divided into group A, those that had a gastric pull-up procedure and group B, those that had a colonic interposition procedure. In total, 50 patients were included; 29 in group A and 21 in group B. Indications included esophageal atresia, caustic ingestion, and infective esophageal stricture. The median age at the time of surgery was 13 months. The mean length of follow-up was 5.2 years. Three patients died giving a mortality rate of 6%; 2 in group A and 1 in group B.In both groups, early postoperative complications included infective complications, such as wound infections, sepsis, and pneumonia (11), anastomotic leak (7), and respiratory complications (7). Late complications included adhesive bowel obstruction (2), anastomotic strictures (4), redundancy (1), and jejunostomy problems (1). Septic complications and anastomotic strictures occurred more frequently in group B. Further surgery was needed in eight patients; this was significantly higher in group B. Full oral feeding was achieved within 6 months in 91.5%. The gastric pull-up and colonic interposition have comparable mortality and outcomes. The colonic interposition was associated with a higher rate of early septic complications, anastomotic strictures, and need for further surgery.


Plasminogen activator inhibitor -1 gene 4G/5G polymorphism in patients with breast cancer.
The plasminogen activator inhibitor-1 (PAI-1) plays an important role in the development of tumor invasion and metastasis. The 4G allele of 4G/5G insertion / deletion polymorphism located in the promoter region 675 bp upstream from the transcription start sequence of PAI-1 gene is responsible for the higher plasma PAI-1 level. The aim of the present study was to evaluate the association between PAI-1 gene 4G / 5G polymorphism and breast cancer. Two groups were investigated: the first group(group 1)was composed of 34 patients with breast cancer, the second group (group 2) consisted of 90 unrelated healthy women without history of malignancy. Genomic DNA isolation was performed from peripheral venous blood by standard phenol-chloroform extraction and polymerase chain reaction of the PAI-1 4G/5G polymorphism was performed. The prevalence of 4G/4G or 4G/5G genotype in group 1 and 2 was 97.1% and 78.8%, respectively. The distribution of the PAI-1 4G/5G genotypes was significantly different between the two groups (p<0.05). We suggest that this polymorphism may contribute to an inherited predisposition to the development of breast cancer, however further studies with larger series from diverse ethnic populations are needed to confirm our results.


Sleep quality and associated factors in Iranian inflammatory bowel disease patients.
Sleep is essential in maintaining health and quality of life. Inflammatory bowel disease (IBD) patients suffer from poor sleep quality. This study aimed to investigate the prevalence of sleep disturbances in Iranian IBD patients as well as the variables which can be attributed to the quality of sleep in IBD patients. Seventy-one patients filled in Pittsburg Sleep Quality Index (PSQI) questionnaire and a sociodemographic questionnaire. Disease activity was assessed by Crohn's Disease (CD) Activity Index and Ulcerative Colitis (UC) Activity Index. Regression analysis was used to identify the association between sociodemographic and disease characteristics with sleep quality. We found that 32.4% of all patients, 23.1% of patients with "in remission to mild" disease, and 66.7% of patients with "moderate" disease, had poor sleep quality. CD patients were more likely to have poor sleep quality comparing UC ones in crude (odds ratio [OR] =2.14; 95% confidence interval [CI] 1.14-4.04) and adjusted (OR = 6.19; 95% CI 1.13, 34.07) models. Patients with good quality of sleep had lower systolic blood pressure and diastolic blood pressure (P = 0.09 and 0.035 respectively). Notable percentage of IBD patients suffer from poor sleep quality even in the remission phase. Treatment of sleep disturbances, especially in CD patients, is recommended in the IBD patient-care program.


BAP Position Statement: Off-label prescribing of psychotropic medication to children and adolescents.
The off-label use of medicines for children and adolescents remains a common and important issue for prescribing practice across child and adolescent psychiatry, paediatrics and primary care. This editorial focusses on psychotropic drug treatment, which plays an essential part in the comprehensive management of a range of child and adolescent psychiatric disorders. Despite a growing evidence base for drug treatment in child and adolescent psychiatric disorders, much psychotropic medication continues to be prescribed off-label (i.e. outside the limits of the marketing authorisation or product license). The reasons for and implications of off-label prescribing, including the potential clinical benefits/risks and medico-legal implications, are often poorly understood by both patients and prescribers. An important unintended consequence of the uncertainties and confusion surrounding the status of off-label prescribing for children and adolescents may be that effective drug treatments are being withheld or underused. This BAP Position Statement aims to clarify these issues, challenge some of the myths surrounding off-label prescribing for children and adolescents and offer practical guidance for prescribers.


Employees' perception of lighting conditions in manufacturing plants: associations with illuminance measurements.
The aims of this study were to evaluate the employees' subjective assessments of different aspects of lighting condition as well as task area illuminance in manufacturing plants. This field study was conducted between March and May 2013, in three packing plants (Saveh, central Iran). Data were collected by questionnaire and measurement of the task area illuminance levels. Data were analysed using contingency coefficient test, Spearman's correlation analysis and non-parametric Friedman tests. The recommended illuminance levels were not met in 46.9% of the work areas. This finding was in agreement with the employees' perception of light level, and with low satisfaction with lighting in the work environment. Adverse effects of lighting condition on job performance, changing posture for better viewing of the work area and eye tiredness were reported as 64%, 33% and 31% of the employees, respectively. Satisfaction with lighting was negatively correlated with the age of respondents (r=-0.229; P<0.010). The employees' satisfaction with lighting was also highly correlated with the employees' subjective assessments of the light level (r=0.779; P<0.001), type of artificial light sources (r=0.591; P<0.001), light colour (r=0.50; P<0.001) and use of daylight (r=0.254; P<0.004). The type of artificial light sources was correlated with job performance (r=0.311, P<0.001) and eye tiredness (r=0.273; P<0.002). The findings highlight the potential usefulness of subjective assessments to supplement objective measures and provide a more holistic approach to lighting design and planning.


A study of clinico-haematological profiles of pancytopenia in children.
We report on the clinico-haematological profile of pancytopenia in children from the Departments of Pediatrics and Pathology, Institute of Medical Sciences, Banaras Hindu University, India, over a period of 30 months. Pancytopenia was defined as: haemoglobin <10 g/dL, absolute neutrophil count 1.5 x 10(9)/L and platelet count <100 x 10(9)/L. A detailed history, clinical examination and haematological parameters were recorded. Bone marrow aspiration and trephine biopsy were carried out in all cases. One hundred and five cases aged 1.5-18 years, with a mean age of 8.6 years, were included in the study. Aplastic anaemia was the most common cause of pancytopenia (43%) followed by acute leukaemia (25%). Infections were the third most common cause of pancytopenia of which kala azar was the most common. Megaloblastic anaemia was seen in 6.7%.


Effects of intraoperative steroid injection on the outcome of pterygium surgery.
To evaluate the effects of intraoperative triamcinolone injection on the outcome of pterygium surgery. This prospective study included 54 eyes with primary nasal pterygia that underwent pterygium surgery with a bare-sclera technique and intraoperative mitomycin C application. Patients were randomized into two groups; the steroid group that received subconjunctival injection of 12 mg triamcinolone acetonide at the end of surgery, and the control group that did not receive such steroid injection. Main outcome measures included presence of conjunctival inflammation at 1 month postoperatively as well as recurrence of pterygium. Twelve-month follow-up was completed in 48 eyes (23 in the steroid group and 25 in the control group). At 1 month postoperatively, different grades of conjunctival inflammation were present in 11 (47.8%) of the steroid group and in 14 (56%) of the control group (P=0.39). For eyes with moderate or severe postoperative inflammation, subconjunctival triamcinolone was injected; these included 6 (26.1%) and 9 (36%) in the steroid and control groups, respectively (P=0.54). During follow-up, surgical area showed fine episcleral vessels without fibrous tissue in 1 (4.3%) of the steroid group and 3 (12.0%) of the control group (P=0.33), which all regressed after triamcinolone injection. Conjunctival recurrence of pterygium was seen in 2 (8.7%) of the steroid group and in 1 (4.0%) of the control group (P=0.47). No eye developed corneal recurrence in either group. In pterygium surgery with a bare-sclera technique and mitomycin C application, intraoperative triamcinolone injection did not significantly reduce postoperative conjunctival inflammation or pterygium recurrence.


Infrapopliteal bypass grafting: use of upper limb vein alone and in autogenous composite grafts.
Thirty-seven consecutive vein grafts to the infrapopliteal arteries are reviewed. Upper limb vein was used in each case, either for the entire graft (27%) or in combination with saphenous vein (73%). The grafts were inserted during a 10-year period and constituted 8.6% of the lower limb grafts carried out during this time. There was one postoperative death (2.7%). The patency rate at 3 years was 58 +/- 9.5% and at 5 years 49.1 +/- 11.5%. Indications for the use of upper limb vein in these situations are considered, and the results are compared with those obtainable with other materials. Upper limb vein can provide a satisfactory arterial substitute in the lower limb, and it may occasionally be the material of choice.


Transition of metabolic phenotypes and risk of subclinical atherosclerosis according to BMI: a prospective study.
The cardiometabolic risk associated with metabolically healthy obesity (MHO) remains the subject of debate. It is unclear whether MHO is a transient condition that affects subclinical atherosclerosis risk. In this study, we aimed to investigate the association of MHO and its transition over time with incident subclinical atherosclerosis. A prospective study was conducted with 6220 Chinese adults who were free of cardiovascular disease (CVD) at baseline. Obesity was defined as BMI ≥25.0 kg/m2. Metabolic health was defined as an individual having fewer than two of the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (NCEP ATP III) criteria for components of the metabolic syndrome (excluding waist circumference). Subclinical atherosclerosis was measured by brachial-ankle pulse wave velocity, pulse pressure and albuminuria, separately or combined. Participants were cross-classified by BMI categories and by metabolic health status and its transition during follow-up. Inverse probability weighted logistic regression models were used to estimate ORs and 95% CIs for subclinical atherosclerosis. The MHO phenotype accounted for 16.3% of the total population and 32.8% of the population with obesity at baseline. Baseline MHO was not significantly associated with incident subclinical atherosclerosis. During a follow-up period of 4.4 years, 46.8% of individuals with MHO developed a metabolically unhealthy status. Those with transient MHO had an increased risk of composite subclinical atherosclerosis compared with those in the metabolically healthy non-obesity reference group (OR 2.52 [95% CI 1.89, 3.36]). A transition from metabolically unhealthy to healthy status was shown to decrease the outcome risk. The relationship between BMI and subclinical atherosclerosis was partly mediated by BP and plasma glucose. MHO is not a stable condition and transient MHO conferred an increased risk of subclinical atherosclerosis, the early stage of CVD. Hence, individuals may benefit from early behavioural or medical management in order to avoid a deterioration of metabolic status and prevent atherosclerosis and CVD.


Psychometric Analysis and Effectiveness of the Psychological Readiness of Injured Athlete to Return to Sport (PRIA-RS) Questionnaire on Injured Soccer Players.
The decision-making process about when an athlete may safely return to training and competition after an injury is a difficult decision. Safe return to training and competition is characterised by physical and psychological readiness to return to the sport. The objectives of this study are (1) to assess the measurement properties of the Psychological Readiness of Injured Athlete to Return to Sport questionnaire (PRIA-RS), and (2) to analyse the effectiveness which the PRIA-RS questionnaire possesses when applied during four consecutive seasons on professional soccer players. One hundred and nine male soccer players from the Albacete Soccer Club (Spain) were involved during four consecutive seasons for the current study: 2012-2013, 2013-2014, 2014-2015 and 2015-2016. Psychometric analysis (validity, reliability, internal consistency and effectiveness) and external psychometric analysis (evaluating measures of patient-reported outcomes (EMPRO)) were confirmed and supported. The main results of the study reveal that the psychometric properties of this questionnaire are optimum for their application in a professional sports context.


Development of a Patient-Reported Outcome Instrument for Patients With Severe Lower Extremity Trauma (LIMB-Q): Protocol for a Multiphase Mixed Methods Study.
A current limitation in the care of patients with severe lower extremity traumatic injuries is the lack of a rigorously developed patient-reported outcome (PRO) instrument specific to lower extremity trauma patients. This mixed methods protocol aims to describe phases I and II of the development of a PRO instrument for lower extremity trauma patients, following international PRO development guidelines. The phase I study follows an interpretive description approach. Development of the PRO instrument begins with identifying the concepts that are important to patients, after which a preliminary conceptual framework is devised from a systematic literature review and used to generate an interview guide. Patients aged 18 years or above with limb-threatening lower extremity traumatic injuries resulting in reconstruction, amputation, or amputation after failed reconstruction will be recruited. The subjects will participate in semistructured, in-depth qualitative interviews to identify all important concepts of interest. The qualitative interview data will be coded with top-level domains, themes, and subthemes. The codes will then be utilized to refine the conceptual framework and generate preliminary items and a set of scales. The preliminary scales will be further refined via a process of conducting cognitive debriefing interviews with lower extremity trauma patients and soliciting expert opinions. Phase III will include a large-scale field test, using Rasch measurement theory to analyze the psychometric properties of the instrument; shortening and finalizing the scales; and determining the reliability, validity, and responsiveness of the instrument. Phases I and II of this study have been funded. Phase I of this study has been completed, and phase II began in January 2019 and is expected to be completed in November 2019. Phase III will begin following the completion of phase II. This protocol describes the initial phases of development of a novel PRO instrument for use in lower extremity trauma patients. DERR1-10.2196/14397.


Assessing subunit dependency of the Plasmodium proteasome using small molecule inhibitors and active site probes.
The ubiquitin-proteasome system (UPS) is a potential pathway for therapeutic intervention for pathogens such as Plasmodium, the causative agent of malaria. However, due to the essential nature of this proteolytic pathway, proteasome inhibitors must avoid inhibition of the host enzyme complex to prevent toxic side effects. The Plasmodium proteasome is poorly characterized, making rational design of inhibitors that induce selective parasite killing difficult. In this study, we developed a chemical probe that labels all catalytic sites of the Plasmodium proteasome. Using this probe, we identified several subunit selective small molecule inhibitors of the parasite enzyme complex. Treatment with an inhibitor that is specific for the β5 subunit during blood stage schizogony led to a dramatic decrease in parasite replication while short-term inhibition of the β2 subunit did not affect viability. Interestingly, coinhibition of both the β2 and β5 catalytic subunits resulted in enhanced parasite killing at all stages of the blood stage life cycle and reduced parasite levels in vivo to barely detectable levels. Parasite killing was achieved with overall low host toxicity, something that has not been possible with existing proteasome inhibitors. Our results highlight differences in the subunit dependency of the parasite and human proteasome, thus providing a strategy for development of potent antimalarial drugs with overall low host toxicity.


[Hormonal dependence of the initial stages of heparin clearance during immobilization stress in the rat].
The 30-min immobilization stress reduces intensity of initial stages of heparin clearance in rats due to increased secretion of ACTH: in blockade of ACTH release the intensity of heparin clearance is not reduced whereas in intensifying the immobilization stress with ACTH administration it is reduced more than in immobilization alone. The stress does not accelerate heparin clearance in adrenalectomized animals. Substitutional administration of hydrocortisone normalizes the clearance in these animals. This suggests a permissive role of glycocorticoids in regulation of heparin clearance intensity. Detainment of heparin in the blood stream in stress is causally related to activation of anticoagulating system and to increased non-enzymatic fibrinolytie activity of the blood due to intensified formation of complex heparin compounds.


Targeted disruption of beta1-integrin in a transgenic mouse model of human breast cancer reveals an essential role in mammary tumor induction.
Despite evidence demonstrating the role of beta1-integrin in the regulation of cancer cell proliferation in vitro, the importance of this cell adhesion receptor during the initiation and progression of epithelial tumors in vivo remains unclear. Here we have used the Cre/LoxP1 recombination system to disrupt beta1-integrin function in the mammary epithelium of a transgenic mouse model of human breast cancer. Using this approach, we show that beta1-integrin expression is critical for the initiation of mammary tumorigenesis in vivo, and for maintaining the proliferative capacity of late-stage tumor cells. These observations provide a direct demonstration that beta1-integrin plays a critical role in both the initiation and maintenance of mammary tumor growth in vivo.


Liquid chromatography used in diagnosis of a rare hemoglobin combination: hemoglobin S/LeporeBoston.
"High-performance" liquid chromatography (HPLC), applied to hemoglobin analysis, is decidely more sensitive and gives better resolution than do routine electrophoretic methods. Here we present a case with a rare double heterozygote hemoglobin S/LeporeBoston, originally diagnosed as homozygous hemoglobin S by routine electrophoretic methods. Using a gradient elution weak cation-exchange HPLC technique, we could separate hemoglobin S and hemoglobin LeporeBoston and make the correct diagnosis. This case demonstrates how HPLC can be a useful adjunct to routine electrophoresis.


Healthy Dietary Patterns and Oxidative Stress as Measured by Fluorescent Oxidation Products in Nurses' Health Study.
Healthy diets may lower oxidative stress and risk of chronic diseases. However, no previous studies examined associations between diet and fluorescent oxidation products (FlOP), a global marker of oxidative stress. We evaluated associations between healthy eating patterns (Alternative Healthy Eating Index (AHEI), Dietary Approach to Stop Hypertension (DASH), and Alternate Mediterranean Diet (aMED)) and FlOP, measured at three excitation/emission wavelengths (FlOP_360, FlOP_320, FlOP_400) from 2021 blood samples collected from 1688 women within the Nurses' Health Study. AHEI, DASH, and aMED scores were significantly positively associated with FlOP_360 and FlOP_320 concentrations (p-trend ≤ 0.04), but not associated with FlOP_400. Among specific food groups that contribute to these diet scores, significantly positive associations were observed with legumes and vegetables for FlOP_360, vegetables and fruits for FlOP_320, and legumes and alcohol for FlOP_400. Inverse associations were observed with nuts, sweets or desserts, and olive oil for FlOP_360, nuts for FlOP_320 and sweets or desserts for FlOP_400 (all p-trend ≤ 0.05). However, FlOP variation due to diet was small compared to overall FlOP variation. In conclusion, AHEI, DASH, and aMED scores were unexpectedly positively, but weakly, associated with FlOP_360 and FlOP_320. However, these findings should be interpreted cautiously as the determinants of FlOP concentrations are not fully understood.


Relationships between Antioxidants and Quality Characteristics from Velvet Antlers of Formosan Sambar Deer.
The quality characteristics of velvet antlers obtained from Formosan sambar deer (Cervus unicolor Swinhoi) (SDVA), harvested from 63 to 81 d during the velvet antler growth period, were evaluated by investigating the relationships between antioxidant levels; including content, activity, and content/activity ratios, and physical properties; including shear force values, color, and Ca content. The hardness of samples from base velvet antler sections increased, and that the color of these samples tended to become reddish-yellow (redder and more yellow), suggesting that the Ca content in the base section of the sample was not ossified yet. Samples from the upper sections of velvet antler showed higher superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) content (3.91 to 1.50 mg/mL, 2.53 to 0.90 mg/mL, and 3.95 to 1.58 mg/mL, respectively) than did samples from the middle and base sections (p<0.05). The activity and content/activity ratios of GPX measured in the upper section were also found to be significantly greater than in the middle and base sections (p<0.05). We further observed that the content and activity of GPX was significantly and negatively correlated with Ca content, shear force values, and the content/activity ratio of this antioxidant (p<0.01). The study findings may serve as a reference index for quality evaluations of velvet antlers of Formosan sambar deer in future.


Approaches to urinary incontinence in a rural population: a comparison of physician assistants, nurse practitioners, and family physicians.
Although urinary incontinence is a challenge and a burden to older patients, many clinicians fail to query older patients about incontinence symptoms or, even when aware of a problem, fail to diagnose the underlying cause or recommend treatment. We wanted to compare the approaches of physician assistants, nurse practitioners, and family physicians to detection, diagnosis, and initial management of urinary incontinence in older adults seen in rural primary care practices. One male and 2 female simulated patients portrayed otherwise healthy patients with urinary incontinence, including urge or obstruction-overflow type for the man, and stress or urge type for the women. The 3 simulated patients saw 3 physician assistants, 3 nurse practitioners, and 3 family physicians each, for a total of 27 visits during which they posed as new patients seeking primary care. Health professionals spontaneously asked about incontinence in only 18 percent of visits (33 percent for physician assistants, 11 percent each for nurse practitioners and family physicians). When incontinence was discussed (spontaneously or by patient prompting), queries were made about potential precipitants (ie, coughing, caffeine consumption) in 63 percent of visits. Questions about other urinary symptoms (eg, dysuria) were asked in 59 percent of visits. Rectal examinations were performed in 68 percent of the male simulated patient's visits but in none of the female simulated patients' visits. No pelvic examinations were performed. No attempts or recommendations were made to measure postvoiding residual volume. Tentative diagnoses were made in 48 percent of visits; some form of therapy was discussed in 52 percent of visits. Asking about incontinence was uncommon, and potentially important questions about precipitants and associated symptoms were often omitted. The providers examined areas potentially relating to incontinence and recommended supplementary assessments and specialized testing infrequently. Commonly, they made diagnoses and offered therapy at the end of an initial visit despite minimal history taking and examinations and lack of any additional assessment or testing.