Lingos, finite state machines, and fast similarity searching.
We apply a recently published method of text-based molecular similarity searching (LINGO) to standard data sets for the purpose of quantifying the accuracy of the approach. Our implementation is based on a pattern-matching finite state machine (FSM) which results in fast search times. The accuracy of LINGO is demonstrated to be comparable to that of a path-based fingerprint and offers a simple yet effective method for similarity searching.


[Combined ROLL and sentinel lymph node: a new strategy in radio-guided surgery].
Radioguided surgery for the treatment of breast cancer is becoming the gold standard for both diagnosis and therapy. The main rule in using ROLL is perfect localization of non-palpable lesions and minimal invasiveness of excision. The same criteria apply to the sentinel lymph node technique. Clinically occult breast lesions, which require an exact histological diagnosis, are most frequently detected as the result of mammographic screening. The authors show that the ROLL technique for histological diagnosis may alter the subdermal lymphatic drainage so that the sentinel node cannot be found at later surgery. The aim of this study was to report the authors' experience with a combined ROLL and sentinel lymph node biopsy technique for the diagnosis and treatment of occult cancer of the breast.


Up-regulation of luciferase gene expression with antisense oligonucleotides: implications and applications in functional assay development.
HeLa Tet-Off cells were transfected transiently as well as stably with a recombinant plasmid (pLuc/705) carrying the luciferase gene interrupted by a mutated human beta-globin intron 2 (IVS2-705). The mutation in the intron causes aberrant splicing of luciferase pre-mRNA, preventing translation of luciferase. However, treatment of the cells with a 2'-O-methyl-oligoribonucleotide targeted to the aberrant splice sites induces correct splicing, restoring luciferase activity. The effects are sequence-specific, depend on the concentration of the oligonucleotide, and can be modulated by the pretreatment of the cell line, Luc/705, with tetracycline. Thus, the cell line provides, among others, a novel functional assay system superior to other procedures that are based on protein down-regulation. In particular, the system would be ideal in assessing the cellular delivery efficiency of antisense oligonucleotides.


Indolylmethanols as Reactants in Catalytic Asymmetric Reactions.
Indolylmethanols have emerged as versatile reactants in catalytic asymmetric reactions because of the facile generation of reactive intermediates from these compounds. The most common indolylmethanols are 3-indolylmethanols and 2-indolylmethanols, and the reaction types applicable to these heterocycles mainly include catalytic enantioselective substitutions and cyclizations. Using indolylmethanols as reactants in catalytic asymmetric reactions enables the enantioselective construction of a variety of indole-containing frameworks.


The Respiratory Exchange Ratio is Associated with Fitness Indicators Both in Trained and Untrained Men: A Possible Application for People with Reduced Exercise Tolerance.
The respiratory exchange ratio (RER) indirectly shows the muscle's oxidative capacity to get energy. Sedentarism, exercise and physically active lifestyles modify it. For that reason, this study evaluates the associations between RER during sub-maximum exercise and other well established fitness indicators (body fat, maximum heart rate, maximum O(2) uptake, workload, and lactate threshold), in physically active trained and untrained men. The RER, O(2) uptake and blood lactate were measured in eight endurance trained and eight untrained men (age, 22.9 ± 4.5 vs. 21.9 ± 2.8 years; body mass, 67.1 ± 5.4 vs. 72.2 ± 7.7 kg; body fat, 10.6 ± 2.4% vs. 16.6 ± 3.8% and maximum O(2) uptake, 68.9 ± 6.3 vs. 51.6 ± 5.8 ml•kg(-1)•min(-1)), during maximum exercise test and during three different sub-maximum exercises at fixed workload: below, within or above the lactate threshold. Endurance trained men presented higher O(2) uptake, lower blood lactate concentrations and lower RER values than those in untrained men at the three similar relative workloads. Even though with these differences in RER, a strong association (p < 0.05) of RER during sub-maximum exercise with the other well established fitness indicators was observed, and both maximum O(2) uptake and lactate threshold determined more than 57% of its variance (p < 0.05). These data demonstrate that RER measurement under sub-maximum exercise conditions was well correlated with other established physical fitness indicators, despite training condition. Furthermore, the results suggest that RER could help obtain an easy approach of fitness status under low exercise intensity and could be utilized in subjects with reduced exercise tolerance.


A novel muscle biopsy clamp yields accurate in vivo sarcomere length values.
The measurement of in vivo muscle sarcomere length facilitates the definition of in vivo muscle functional properties and comparison of muscle designs amongst functional muscle groups. In vivo sarcomere lengths are available for just a handful of human muscles, largely due to the technical challenges associated with their measurement. The purpose of this report was to develop and test a muscle biopsy clamp that can quickly and accurately measure in vivo muscle sarcomere length. To test the device, muscle biopsies (n=23) were removed from the tibialis anterior muscles of New Zealand White rabbits immediately after sarcomere length measurements were made using laser diffraction. The muscle biopsy contained within the clamp was immediately fixed in Formalin for subsequent sarcomere length measurement. Comparisons of clamp-based and diffraction-based sarcomere lengths demonstrated excellent agreement between the two techniques, especially when the biopsy was obtained at relatively long lengths (above 2.6 microm). Given the intraoperative speed and simplicity of this technique and the relatively low-cost of the biopsy clamp, this method of measuring muscle sarcomere length should help investigators generate much-needed in vivo muscle structural and functional data.


Support Issues and Nursing Doctoral Students' Intent to Leave.
The purpose of this study was to explore, analyze, and describe nursing doctoral students' perceptions of factors that impacted them during their doctoral program. Answers to an open-ended question, in a national descriptive survey study of nursing doctoral students across the United States, were analyzed using Colaizzi procedural methods as a guideline. The content analysis identified issues unique to doctor of nursing practice (DNP) students that related to lack of clarity for the DNP degree and student concerns within academic settings. Recommendations include having DNP faculty on DNP project committees and standardization of guidelines to ensure rigor and consistency.


Environmental and socio-economic impacts of new plant breeding technologies: A case study of root chicory for inulin production.
In Europe, root chicory and other plants are cultivated for their prebiotic food fiber, inulin, which boosts the growth of beneficial gut bacteria and stimulates the human immune system. CHIC, a H2020 project, develops new chicory variants which produce more and reported to be healthier inulin as well as medicinal terpenes. This paper presents an environmental and socio-economic assessment of the whole value chain of the new chicory variants and their derived products using a case study based in the Netherlands. Two scenarios based on new chicory variants using new plant breeding technologies (NPBT) are analyzed and impacts thereof are compared to the reference scenario; the current commercial inulin process from conventional chicory. Both scenarios show higher inulin content, but the inulin adsorption process differs. While one aims to optimize inulin yield, the other one explores the potential of a multipurpose use, yielding inulin and health beneficial terpenes. Methodologically, we employ multi-regional input-output (MRIO) analysis to estimate additional economic benefits, added value and job creation, while by means of life cycle assessment (LCA) effects on greenhouse gas (GHG) emissions and primary energy demand are derived. Both methods, MRIO and LCA, are well suited to analyze the raised issues and draw on the same data. Generally, the results highlight the importance of inulin production at a national and EU-level in the reference scenario. In case of the two scenarios, we find that the related socio-economic impacts are much higher than in the reference scenario and thus highlight their ability to boost economic activity and increase competiveness of the EU, i.e. over 80% of the generated value added stays in the EU. In terms of environmental impacts, the two scenarios show lower GHG emissions and primary energy demand due to the higher efficiencies of the process in the scenarios compared to the reference inulin process. Additionally, regarding the goal of climate neutral production, we find that the majority of GHG emissions stem from the electricity mix and natural gas demand. Replacing these sources of energy with more renewable ones will contribute to this goal.


Strategies of rehabilitation in the Day Hospital.
A correct and fruitful rehabilitation strategy should consider the following points: (1) the achievement of a psychic equilibrium with a reduction of the psychopathological symptoms; (2) the evaluation of the psychological handicap; (3) a profile of the abilities of the subject that undergoes rehabilitation. You can easily achieve the first two points during hospitalization. The third point can be achieved through the study of the interaction of the subject with his or her environment. This study can be carried out in the Day Hospital where, through groups of psychotherapy, family counseling and various sociotherapeutic activities, it is possible to increase and improve the ability of the subject to relate with the others and to cope adequately with his or her environment. In this paper we present the philosophy and the therapeutic strategies of the Day Hospital of the Psychiatric Department of the University of Milan.


Creatine kinase isoenzymes in cerebrospinal fluid in a case of brain damage.
We present a case of a 11/2-year-old boy with toxic enteritis, consecutive consumption coagulopathy, and sever brain damage. During the acute phase we found high activity of the BB isoenzyme of creatine kinase in cerebrospinal fluid, but not in the serum. Isoenzyme MM could also be found in the spinal fluid (37.9% of the total activity). We conclude that analysis for creatine kinase isoenzymes in spinal fluid is of clinical importance.


Zika virus: A public health threat.
The resurgence of Zika virus as public health emergency of an international concern with increased incidence of microcephaly has drawn attention of scientific community for its detailed understanding with regard to virus evolution, epidemiology, geographical spread, pathogenesis, etc. The scope of the present review is to discuss the detailed updated information in respect of Zika virus evolution since its inception.


Aggregation of prion protein with insertion mutations is proportional to the number of inserts.
Mutation in the prion gene, PRNP, accounts for approx. 10-15% of human prion diseases. However, little is known about the mechanisms by which a mutant prion protein (PrP) causes disease. We compared the biochemical properties of a wild-type human prion protein, rPrP(C) (recombinant wild-type PrP), which has five octapeptide-repeats, with two recombinant human prion proteins with insertion mutations, one with three more octapeptide repeats, rPrP(8OR), and the other with five more octapeptide repeats, rPrP(10OR). We found that the insertion mutant proteins are more prone to aggregate, and the degree and kinetics of aggregation are proportional to the number of inserts. The octapeptide-repeat and alpha-helix 1 regions are important in aggregate formation, because aggregation is inhibited with monoclonal antibodies that are specific for epitopes in these regions. We also showed that a small amount of mutant protein could enhance the formation of mixed aggregates that are composed of mutant protein and wild-type rPrP(C). Accordingly, rPrP(10OR) is also more efficient in promoting the aggregation of rPrP(C) than rPrP(8OR). These findings provide a biochemical explanation for the clinical observations that the severity of the disease in patients with insertion mutations is proportional to the number of inserts, and thus have implications for the pathogenesis of inherited human prion disease.


Advantages of Foxp3(+) regulatory T cell depletion using DEREG mice.
Several mechanisms enable immunological self-tolerance. Regulatory T cells (Tregs) are a specialized T cell subset that prevents autoimmunity and excessive immune responses, but can also mediate detrimental tolerance to tumors and pathogens in a Foxp3-dependent manner. Genetic tools exploiting the foxp3 locus including bacterial artificial chromosome (BAC)-transgenic DEREG mice have provided essential information on Treg biology and the potential therapeutic modulation of tolerance. In DEREG mice, Foxp3(+) Tregs selectively express eGFP and diphtheria toxin (DT) receptor, allowing for the specific depletion of Tregs through DT administration. We here provide a detailed overview about important considerations such as DT toxicity, which affects any mouse strain treated with DT, and Treg rebound after depletion. Additionally, we point out the specific advantages of BAC-transgenic DEREG mice including their suitability to study organ-specific autoimmunity such as type I diabetes. Moreover, we discuss recent insights into the role of Tregs in viral infections. In summary, DEREG mice are an important tool to study Treg-mediated tolerance and its therapeutic circumvention.


Adolescents' Perceptions About Cancer and Preferences for Cancer Education.
Cancer is one of the leading causes of death within the United States. Adolescence remains a critical stage of development in which new cognitive skills and lifestyle factors related to cancer are acquired. It is critical to understand adolescents' knowledge of cancer and preferences for receiving clear and easy-to-comprehend information about cancer and cancer prevention. The purpose of this study was to explore adolescents' perceptions of cancer and cancer risk factors and identify their preferences for receiving cancer prevention education. The survey used in this study was based on the Health Belief Model framework, the Youth Risk Behavior Survey, and the Cancer, Clear & Simple (CC&S) Curriculum. The survey evaluated adolescents' knowledge, attitudes, and health-related behaviors regarding cancer and cancer prevention among adolescents. Three Midwestern middle and high schools were recruited, and 235 students completed the survey. Survey responses were analyzed using descriptive statistics. Study participants had some basic knowledge of cancer; however, only 66% recognized that individuals have control over their risk of developing cancer. Participants (95%) reported that cancer prevention was important, yet only 37.3% stated they knew how to lower their risk of getting cancer and 50% indicated that they currently try to make decisions that lower their chances of getting cancer. About 82% stated that an educational game would be a useful tool to learn about cancer and prevention. Study findings indicate the need for an increase in adolescent knowledge of cancer, specifically how they can control their cancer risk, and prevention strategies through development of game-based education to teach adolescents about cancer-related topics.


BMP signalling regulates the pre-implantation development of extra-embryonic cell lineages in the mouse embryo.
Pre-implantation development requires the specification and organization of embryonic and extra-embryonic lineages. The separation of these lineages takes place when asymmetric divisions generate inside and outside cells that differ in polarity, position and fate. Here we assess the global transcriptional identities of these precursor cells to gain insight into the molecular mechanisms regulating lineage segregation. Unexpectedly, this reveals that complementary components of the bone morphogenetic protein (BMP) signalling pathway are already differentially expressed after the first wave of asymmetric divisions. We investigate the role of BMP signalling by expressing dominant negative forms of Smad4 and Bmpr2, by downregulating the pathway using RNA interference against BMP ligands and by applying three different BMP inhibitors at distinct stages. This reveals that BMP signalling regulates the correct development of both extra-embryonic lineages, primitive endoderm and trophectoderm, but not the embryonic lineage, before implantation. Together, these findings indicate multiple roles of BMP signalling in the early mouse embryo.


Characterization of the Synergistic Antioxidant Activity of Epigallocatechin Gallate (EGCG) and Kaempferol.
Epigallocatechin gallate (EGCG) and kaempferol exhibit cellular antioxidant activity; however, their interactive effects in terms of antioxidant actions and underlying mechanisms remain unclear. In this study, their cytoprotective effects were examined against 2,2-azobis (2-amidinopropane) dihydrochloride solution (ABAP)-induced oxidative stress in HepG2 cells. The results showed that the median effective dose (EC50) of the EGCG and kaempferol (6:1.5, c/c) combination was 3.4 ± 0.1 μg/mL, with a combination index (CIavg) value of 0.54, which represented a significant synergistic effect. Further experiments proved that the combined pretreatment with EGCG and kaempferol exerted protective effects by suppressing reactive oxygen species (ROS) generation, upregulating cellular antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px)) in a dose-dependent manner. The mechanism of synergistic antioxidant effects of EGCG combined with kaempferol may be due to the up-regulation of higher antioxidant enzyme activities that improve the antioxidant capacities and balance the cell oxidative stress. The synergistic antioxidant effect of EGCG and kaempferol can provide a theoretical basis for the development of formulas of functional food ingredients.


HIV and antiretroviral treatment knowledge gaps and psychosocial burden among persons living with HIV in Lima, Peru.
This study aims to describe knowledge on HIV and antiretroviral (ARV) treatment and psychosocial factors among people living with HIV (PLWH) in Lima, Perú, to explore characteristics associated to this knowledge, and determine its impact on sustained viral suppression. A cross-sectional survey was conducted among 171 PLWH at the largest referral health care center in Lima. The psychosocial factors measured were depression, risk of alcoholism, use of illegal drugs and disclosure. A participant had "poor knowledge" when less than 80% of replies were correct. Sustained viral suppression was defined as two consecutive viral loads under 50 copies/mL. A total of 49% and 43% had poor HIV and ARV knowledge respectively; 48% of the study population screened positive for depression and 27% reported feeling unsupported by the person they disclosed to. The largest gaps in HIV and ARV knowledge were among 98 (57%) that did not recognize that HIV increased the risk of cancer and among 57 (33%) participants that did not disagree with the statement that taking a double dose of ARV if they missed one. Moderate depression was significantly associated to poor HIV and ARV knowledge. Non-disclosure and being on ARVs for less than 6 months were associated with not achieving sustained viral suppression. Our findings highlight important HIV and ARV knowledge gaps of PLWH and a high burden of psychosocial problems, especially of depression, among PLWH in Lima, Peru. Increasing knowledge and addressing depression and disclosure could improve care of PLWH.


An ab Initio Study of Intramolecular Hydrogen Bondings in α-Hydroxy Ketomethylene Dipeptide Isostere.
Ab initio calculations of the representative α-hydroxy ketomethylene dipeptide isostere (2S,5S)-5-amino-2-hydroxy-4-oxohexanoic acid (1) are described. All calculations including full geometry optimizations were performed at the MP2/6-31G* level. In the gas phase, 12 low-energy conformers are located by minimizing geometries assembled from stable molecular fragments. Among these conformers, six structurally similar conformers, in which the 2-hydroxyl group forms hydrogen bondings with both the O atom of the 4-carbonyl group in 1,3-fashion and the O atom of 1-carboxylic acid in 1,2-fashion simultaneously, are found to be particularly stable. Thus, the conformational preference of 1 appears to be governed by arrangements and strength of intramolecular hydrogen bondings. To examine conformational natures of 1 in solutions more accurately, we corrected the thermochemical properties and carried out self-consistent reaction field calculations. Going from the gas phase to solutions, the basic features of the conformational preferences in 1 also appear to be maintained in solutions including a highly polar aqueous medium, despite slight changes in the population of each conformer.


Genome-wide landscape of position effects on heterogeneous gene expression in Saccharomyces cerevisiae.
Integration of heterogeneous genes is widely applied in synthetic biology and metabolic engineering. However, knowledge about the effect of integrative position on gene expression remains limited. We established a genome-wide landscape of position effect on gene expression in Saccharomyces cerevisiae. The expression cassette of red fluorescence protein (RFP) gene was constructed and inserted at 1044 loci, which were scattered uniformly in the yeast genome. Due to the different integrative loci on the genome, the maximum relative intensity of RFP is more than 13-fold over the minimum. Plots of the number of strains to RFP relative intensity showed normal distribution, indicating significant position effect on gene expression in yeast. Furthermore, changing the promoters or reporter genes, as well as carbon sources, revealed little consequences on reporter gene expression, indicating chromosomal location is the major determinant of reporter gene expression. We have examined the position effects to integration genes expression in large number loci around whole genome in S. cerevisiae. The results could guide the design of integration loci for exogenous genes and pathways to maximize their expression in metabolic engineering.


Gut metabolites of anthocyanins, gallic acid, 3-O-methylgallic acid, and 2,4,6-trihydroxybenzaldehyde, inhibit cell proliferation of Caco-2 cells.
Gut microflora metabolize anthocyanins to phenolic acids and aldehydes. These metabolites may explain the relationship between anthocyanin consumption and reduced incidence of colon cancer. Here, all six major metabolites, along with a Cabernet Sauvignon anthocyanin extract, were incubated with Caco-2 cells at concentrations of 0-1000 microM over 72 h to determine effects on cell proliferation and for 24 h to assess cytotoxicity effects and at 140 microM for 24 h to measure induction of apoptosis. These measurements were based on colorimetric methods. Gallic acid and 3-O-methylgallic acid inhibited cell proliferation and lacked cytotoxicity at low concentrations. The aldehyde metabolite and anthocyanin extract also inhibited cell proliferation at low concentrations and had low cytotoxicity at a wide range of concentrations. Of the four substances that effectively reduced cell proliferation, the aldehyde was the best inducer of apoptosis. In addition, these same four treatments degraded quickly in growth media, suggesting the involvement of subsequent oxidation products in the reduction of cell viability. These results indicate that the anthocyanin microfloral metabolites gallic acid, 3-O-methylgallic acid, and 2,4,6-trihydroxybenzaldehyde reduce cell proliferation in Caco-2 cells more effectively than anthocyanins and may offer protection against colon cancer after their formation in the gut.


Methane to Chloromethane by Mechanochemical Activation: A Selective Radical Pathway.
State-of-the-art processes to directly convert methane into CH3Cl are run under corrosive conditions and typically yield a mixture of chloromethanes requiring subsequent separation. We report a mechanochemical strategy to selectively convert methane to chloromethane under overall benign conditions, employing trichloroisocyanuric acid (TCCA) as a cheap and noncorrosive solid chlorinating agent. TCCA is shown to release active chlorine species upon milling with Lewis acids such as alumina and ceria to functionalize methane at moderate temperatures (<150 °C). A thorough parameter optimization led to a maximum methane chlorination rate of 0.8 μmol(CH4,conv) (g(catalyst) s)-1. Findings were compared to the thermal reaction of methane with TCCA and evidenced that mechanochemical activation permitted significantly lower reaction temperatures (90 vs 200 °C) at a drastically improved CH3Cl selectivity (95% vs 66% at 30% conversion). Considering the characterization of the interaction between TCCA and Lewis acids as well as the in-depth analysis of byproducts, we suggest a plausible reaction mechanism and a possible regeneration of the chlorinating agent.


The challenge of measuring community values in ways appropriate for setting health care priorities.
The move from a notion that community values ought to play a role in health care decision making to the creation of health care policies that in some way reflect such values is a challenging one. No single method will adequately measure community values in a way appropriate for setting health care priorities. Consequently, multiple methods to measure community values should be employed, thereby allowing the strengths and weaknesses of the various methods to complement each other. A preliminary research agenda to bring together empirical research on community values with more traditional research on health care ethics is outlined, with the goal of identifying and measuring acceptable community values that are relatively consistent across measurement methods and, ultimately, developing ways to incorporate these values into health care priority decision making.


Cytoplasmic tail phosphorylation of the alpha-factor receptor is required for its ubiquitination and internalization.
G protein-coupled (GPC) receptors are phosphorylated in response to ligand binding, a modification that promotes receptor desensitization or downregulation. The alpha-factor pheromone receptor (Ste2p) of Saccharomyces cerevisiae is a GPC receptor that is hyperphosphorylated and ubiquitinated upon binding alpha-factor. Ubiquitination triggers Ste2p internalization into the endocytic pathway. Here we demonstrate that phosphorylation of Ste2p promotes downregulation by positively regulating ubiquitination and internalization. Serines and a lysine are essential elements of the Ste2p SINNDAKSS internalization signal that can mediate both constitutive and ligand-stimulated endocytosis. The SINNDAKSS serines are required for receptor phosphorylation which, in turn, facilitates ubiquitination of the neighboring lysine. Constitutive phosphorylation is required to promote constitutive internalization, and is also a prerequisite for ligand-induced phosphorylation at or near the SINNDAKSS sequence. Mutants defective in yeast casein kinase I homologues are unable to internalize alpha-factor, and do not phosphorylate or ubiquitinate the receptor, indicating that these kinases play a direct or indirect role in phosphorylating the receptor. Finally, we provide evidence that the primary function of phosphorylation controlled by the SINNDAKSS sequence is to trigger receptor internalization, demonstrating that phosphorylation-dependent endocytosis is an important mechanism for the downregulation of GPC receptor activity.


Dexmedetomidine Inhibits Phenylephrine-induced Contractions via Alpha-1 Adrenoceptor Blockade and Nitric Oxide Release in Isolated Rat Aortae.
The goal of this in vitro study was to examine the effect of the alpha-2 adrenoceptor agonist dexmedetomidine on phenylephrine (alpha-1 adrenoceptor agonist)-induced contraction in isolated rat aortae and to elucidate the associated cellular mechanisms, with a particular focus on alpha-1 adrenoceptor antagonism. Dexmedetomidine dose-response curves were generated in isolated endothelium-intact and endothelium-denuded rat aortae precontracted with phenylephrine or 5-hydroxytryptamine. Endothelium-denuded aortic rings were pretreated with either dexmedetomidine or the reversible alpha-1 adrenoceptor antagonist phentolamine, followed by post-treatment with the irreversible alpha-1 adrenoceptor blocker phenoxybenzamine. Control rings were treated with phenoxybenzamine alone. All rings were repeatedly washed with Krebs solution to remove all pretreatment drugs, including phenoxybenzamine, phentolamine and dexmedetomidine. Phenylephrine dose-response curves were then generated. The effect of rauwolscine on the dexmedetomidine-mediated change in phenylephrine-induced endothelial nitric oxide synthase (eNOS) phosphorylation in human umbilical vein endothelial cells was examined using western blotting. The magnitude of the dexmedetomidine-mediated inhibition of phenylephrine-induced contraction was higher in endothelium-intact aortae than in endothelium-denuded aortae or endothelium-intact aortae treated with Nω-nitro-L-arginine methyl ester. However, dexmedetomidine did not significantly alter 5-hydroxytryptamine-induced contraction. In further experiments, prazosin attenuated dexmedetomidine-induced contraction. Additionally, pretreatment with either dexmedetomidine plus phenoxybenzamine or phentolamine plus phenoxybenzamine produced greater phenylephrine-induced contraction than phenoxybenzamine alone, suggesting that dexmedetomidine protects aortae from the alpha-1 adrenoceptor blockade induced by phenoxybenzamine. Rauwolscine attenuated the dexmedetomidine-mediated enhancement of phenylephrine-induced eNOS phosphorylation. Taken together, these results suggest that dexmedetomidine attenuates phenylephrine-induced contractions via alpha-1 adrenoceptor blockade and endothelial nitric oxide release in the isolated rat aorta.


Properties of a rat monoclonal antibody reactive with both the mannan of Candida species and the O-antigen 6,7 polysaccharide of serogroup C1 salmonellae.
Monoclonal antibody MASC1-MR9 of isotype immunoglobulin M was generated in LOU/C rats by immunization with heat-killed Salmonella thompson (serogroup C1, O-antigen 6,7 lipopolysaccharide) bacteria which had been further enriched for O-antigen by coating with homologous lipopolysaccharide. Eight monoclonal antibodies were selected by screening against the lipopolysaccharide of S. thompson. In a subsequent test against Candida mannan, only one antibody, MASC1-MR9, was reactive. Immunofluorescence microscopy of various yeasts and bacteria with MASC1-MR9 showed that this antibody bound to the surfaces of 142 of 148 Candida strains of 10 different species tested but failed to bind to the surface of Saccharomyces cerevisiae. The Candida strains which failed to bind MASC1-MR9 were all five strains of Candida krusei and the single strain of Candida utilis tested. Several (11 of 33) Salmonella strains belonging to serogroup C1 reacted with this antibody, as expected; however, 1 of 11 strains of Morganella morganii and 4 of 64 strains of Escherichia coli were also reactive. Serum-free supernatants of MASC1-MR9 agglutinated S. thompson, one strain of Salmonella choleraesuis, and 109 of 110 Candida strains tested. The immunochemical properties of MASC1-MR9 as studied by enzyme-linked immunosorbent inhibition assays show that it recognizes an epitope present in the mannan of Candida species as well as in the O-6,7 antigen of Salmonella species.


[Glaucoma, tonometry and anaesthesia (author's transl)].
Definition and classification of glaucomas. Indication of drugs contraindicated in glaucomas. Discussion of the problem of tonometry in anaesthesia in congenital glaucoma suspicion with special regard to factors influencing intraocular pressure. Discussion of correlation between intraocular pressure and systemic blood pressure. Report on own experiences with Ketamine and discussion of advantage and disadvantage of the drug for tonometry in anaesthesia in infants. Explanation of the method of impression--and applanation tonometry with regard to Glaucotest, a new borderline--tonometer suitable for the anaesthesiologist.


Striatal and extrastriatal dysfunction in Parkinson's disease with dementia: a 6-[18F]fluoro-L-dopa PET study.
We investigated the relative differences in dopaminergic function through the whole brain in patients with Parkinson's disease without dementia (PD) and with dementia (PDD) using 6-[18F]fluoro-L-dopa (18F-dopa) PET and a voxel-by-voxel analysis. The 10 PD and 10 PDD patients were equivalently disabled, having mean scores of 3.2 +/- 0.6 and 3.2 +/- 0.7, respectively, on the Hoehn and Yahr rating scale. 18F-dopa influx constant (Ki) images of those patients and 15 normal age-matched subjects were transformed into standard stereotactic space. The significant differences between the groups (expressed in mean regional Ki values) were localized with statistical parametric mapping (SPM) on a voxel-by-voxel basis. Compared with the normal group, SPM localized declines of the 18F-dopa Ki bilaterally in the putamen, the right caudate nucleus and the left ventral midbrain for the PD group (P < 0.01, corrected). Compared with the normal group, the PDD group showed reduced 18F-dopa Ki bilaterally in the striatum, midbrain and anterior cingulate area (P < 0.01, corrected). A relative difference in 18F-dopa uptake between PD and PDD was the bilateral decline in the anterior cingulate area and ventral striatum and in the right caudate nucleus in the PDD group (P < 0.001, corrected). Accordingly, we conclude that dementia in PD is associated with impaired mesolimbic and caudate dopaminergic function.


Alternative complement pathway in the pathogenesis of disease mediated by anti-neutrophil cytoplasmic autoantibodies.
Clinical and experimental data indicate that anti-neutrophil cytoplasmic autoantibodies (ANCAs) cause glomerulonephritis and vasculitis. Here we report the first evidence that complement is an important mediator of ANCA disease. Transfer of anti-myeloperoxidase (MPO) IgG into wild-type mice or anti-MPO splenocytes into immune-deficient mice caused crescentic glomerulonephritis that could be completely blocked by complement depletion. The role of specific complement activation pathways was investigated using mice with knockout of the common pathway component C5, classic and lectin binding pathway component C4, and alternative pathway component factor B. After injection of anti-MPO IgG, C4-/- mice developed disease comparable with wild-type disease; however, C5-/- and factor B-/- mice developed no disease. To substantiate a role for complement in human ANCA disease, IgG was isolated from patients with myeloperoxidase ANCA (MPO-ANCA) or proteinase 3 ANCA (PR3-ANCA) and from controls. Incubation of MPO-ANCA or PR3-ANCA IgG with human neutrophils caused release of factors that activated complement. IgG from healthy controls did not produce this effect. The findings suggest that stimulation of neutrophils by ANCA causes release of factors that activate complement via the alternative pathway, thus initiating an inflammatory amplification loop that mediates the severe necrotizing inflammation of ANCA disease.


Robot-Assisted Laparoscopic Donor Nephrectomy vs Standard Laparoscopic Donor Nephrectomy: A Prospective Randomized Comparative Study.
The goal of this randomized controlled trial was to compare the outcomes of robot-assisted laparoscopic donor nephrectomy (RDN) with standard laparoscopic donor nephrectomy (LDN). Forty-five voluntary kidney donors (27 for right subgroup and 18 for left subgroup) who met inclusion and exclusion criteria were randomized into 2 groups, RDN and LDN in 1:2 ratio. Primary endpoints were visual analogue scale (VAS) pain scores, analgesic requirement, and hospital stay of donors. Secondary endpoints were donor's intraoperative and postoperative parameters, graft outcomes, and donor surgeon's difficulty scores. All procedures were completed without any intraoperative complications. VAS pain scores at 6, 24, and 48 hours (p = 0.00), analgesic requirement (p = 0.00), and hospital stay (p = 0.00) were less in RDN than in LDN. Longer graft arterial length could be preserved with robotic approach on right side (p = 0.03) but not on left side (p = 0.77). The RDN group required more number of ports (p = 0.00), longer retrieval time (p = 0.00), and warm ischemia time (WIT) (p = 0.01). Total operative time (p = 0.14), hemoglobin drop (p = 0.97), postoperative donor complications (p = 0.97), and the recipient estimated glomerular filtration rate at 9 months (p = 0.64) were similar in both groups. Difficulty scores of console surgeon were less in most steps on right side but not on left side. Patient-side surgeon in RDN had higher difficulty scores for retrieval. RDN is safe and is associated with better morbidity profile than LDN. Robotic approach provides technical ease and facilitates preservation of longer length of renal artery on right side. Left RDN is associated with longer WIT; however, this does not translate into poor graft outcome.


Rapid Assembly of Magnetoplasmonic Photonic Arrays for Brilliant, Noniridescent, and Stimuli-Responsive Structural Colors.
There are usually trade-offs between maximizing the color saturation and brightness and minimizing the angle-dependent effect in structural colors. Here, a magnetic field-induced assembly for the rapid formation of scalable, uniform amorphous photonic arrays (APAs) featuring unique structural colors is demonstrated. The magnetic field plays a fundamental role in photonic film formation, making this assembly technology versatile for developing structural color patterns on arbitrary substrates. The synergistic combination of surface plasmonic resonance of the Ag core and broadband light absorption of high refractive index (RI) Fe3 O4 shell in hybrid magnetoplasmonic nanoparticles (MagPlas NPs) enables breaking the trade-offs to produce brilliant, noniridescent structural colors with high tunability and responsiveness. These features enable the fabrication of various types of highly sensitive and reliable colorimetric sensors for naked-eye detection without sophisticated instruments. Furthermore, large-scale structural color patterns are effortlessly achieved, demonstrating the high potential of the present approach for full-spectrum displays, active coatings, and rewritable papers.


Traffic, air pollution, and health.
This paper discusses emerging information on exposure to air pollution from traffic and health and raises two key questions with regard to the impact of traffic on public health: 1) is there sufficient evidence to infer that traffic is causing adverse health effects, and; 2) what is the magnitude of the adverse impact of traffic on health? Topics addressed include characterization of exposure to traffic-related pollutants; the findings of epidemiological studies to date; and investigative approaches and the related challenges - including bias, model-based conclusions, and sample size issues--in characterizing the health effects of air pollution from traffic. Also considered are the known health effects of two of the major pollutants produced by vehicle exhaust--particles and ozone. The evidence points to traffic as a threat to public health that will be managed with great difficulty; however, more research is needed to refine our understanding of the health consequences of traffic exposures and as a basis for formulating mitigation policies. This paper discusses emerging information on exposure to air pollution from traffic and health and raises two key questions with regard to the impact of traffic on public health: 1) is there sufficient evidence to infer that traffic is causing adverse health effects, and; 2) what is the magnitude of the adverse impact of traffic on health? Topics addressed include characterization of exposure to traffic-related pollutants; the findings of epidemiological studies to date; and investigative approaches and the related challenges - including bias, model-based conclusions, and sample size issues - in characterizing the health effects of air pollution from traffic. Also considered are the known health effects of two of the major pollutants produced by vehicle exhaust - particles and ozone. The evidence points to traffic as a threat to public health that will be managed with great difficulty; however, more research is needed to refine our understanding of the health consequences of traffic exposures and as a basis for formulating mitigation policies.


Site-specific loss of acetylation upon phosphorylation of histone H3.
Post-translational modification of histones is a central aspect of gene regulation. Emerging data indicate that modification at one site can influence modification of a second site. As one example, histone H3 phosphorylation at serine 10 (Ser(10)) facilitates acetylation of lysine 14 (Lys(14)) by Gcn5 in vitro (, ). In vivo, phosphorylation of H3 precedes acetylation at certain promoters. Whether H3 phosphorylation globally affects acetylation, or whether it affects all acetylation sites in H3 equally, is not known. We have taken a genetic approach to this question by mutating Ser(10) in H3 to fix either a negative or a neutral charge at this position, followed by analysis of the acetylation states of the mutant histones using site-specific antibodies. Surprisingly, we find that conversion of Ser(10) to glutamate (S10E) or aspartate (S10D) causes almost complete loss of H3 acetylation at lysine 9 (Lys(9)) in vivo. Acetylation of Lys(9) is also significantly reduced in cells bearing mutations in the Glc7 phosphatase that increase H3 phosphorylation levels. Mutation of Ser(10) in H3 and the concomitant loss of Lys(9) acetylation has minimal effects on expression of a Gcn5-dependent reporter gene. However, synergistic growth defects are observed upon loss of GCN5 in cells bearing H3 Ser(10) mutations that are reminiscent of delays in G(2)/M progression caused by combined loss of GCN5 and acetylation site mutations. Together these results demonstrate that H3 phosphorylation directly causes site-specific and opposite changes in acetylation levels of two residues within this histone, Lys(9) and Lys(14), and they highlight the importance of these histone modifications to normal cell functions.


Metabolism of vertebrate amino sugars with N-glycolyl groups: incorporation of N-glycolylhexosamines into mammalian glycans by feeding N-glycolylgalactosamine.
The outermost positions of mammalian cell-surface glycans are predominantly occupied by the sialic acids N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). To date, hydroxylation of CMP-Neu5Ac resulting in the conversion into CMP-Neu5Gc is the only known enzymatic reaction in mammals to synthesize a monosaccharide carrying an N-glycolyl group. In our accompanying paper (Bergfeld, A. K., Pearce, O. M., Diaz, S. L., Pham, T., and Varki, A. (2012) J. Biol. Chem. 287, jbc.M112.363549), we report a metabolic pathway for degradation of Neu5Gc, demonstrating that N-acetylhexosamine pathways are tolerant toward the N-glycolyl substituent of Neu5Gc breakdown products. In this study, we show that exogenously added N-glycolylgalactosamine (GalNGc) serves as a precursor for Neu5Gc de novo biosynthesis, potentially involving seven distinct mammalian enzymes. Following the GalNAc salvage pathway, UDP-GalNGc is epimerized to UDP-GlcNGc, which might compete with the endogenous UDP-GlcNAc for the sialic acid biosynthetic pathway. Using UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase-deficient cells, we confirm that conversion of GalNGc into Neu5Gc depends on this key enzyme of sialic acid biosynthesis. Furthermore, we demonstrate by mass spectrometry that the metabolic intermediates UDP-GalNGc and UDP-GlcNGc serve as substrates for assembly of most major classes of cellular glycans. We show for the first time incorporation of GalNGc and GlcNGc into chondroitin/dermatan sulfates and heparan sulfates, respectively. As demonstrated by structural analysis, N-glycolylated hexosamines were found in cellular gangliosides and incorporated into Chinese hamster ovary cell O-glycans. Remarkably, GalNAc derivatives altered the overall O-glycosylation pattern as indicated by the occurrence of novel O-glycan structures. This study demonstrates that mammalian N-acetylhexosamine pathways and glycan assembly are surprisingly tolerant toward the N-glycolyl substituent.


Crystal structure of the R-protein of the multisubunit ATP-dependent restriction endonuclease NgoAVII.
The restriction endonuclease (REase) NgoAVII is composed of two proteins, R.NgoAVII and N.NgoAVII, and shares features of both Type II restriction enzymes and Type I/III ATP-dependent restriction enzymes (see accompanying paper Zaremba et al., 2014). Here we present crystal structures of the R.NgoAVII apo-protein and the R.NgoAVII C-terminal domain bound to a specific DNA. R.NgoAVII is composed of two domains: an N-terminal nucleolytic PLD domain; and a C-terminal B3-like DNA-binding domain identified previously in BfiI and EcoRII REases, and in plant transcription factors. Structural comparison of the B3-like domains of R.NgoAVII, EcoRII, BfiI and the plant transcription factors revealed a conserved DNA-binding surface comprised of N- and C-arms that together grip the DNA. The C-arms of R.NgoAVII, EcoRII, BfiI and plant B3 domains are similar in size, but the R.NgoAVII N-arm which makes the majority of the contacts to the target site is much longer. The overall structures of R.NgoAVII and BfiI are similar; however, whilst BfiI has stand-alone catalytic activity, R.NgoAVII requires an auxiliary cognate N.NgoAVII protein and ATP hydrolysis in order to cleave DNA at the target site. The structures we present will help formulate future experiments to explore the molecular mechanisms of intersubunit crosstalk that control DNA cleavage by R.NgoAVII and related endonucleases.


Clinical and prostate multiparametric magnetic resonance imaging findings as predictors of general and clinically significant prostate cancer risk: A retrospective single-center study.
To evaluate the predictive values of Prostate Imaging Reporting and Data System version 2 (PI-RADS v2), prostate-specific antigen (PSA) level, PSA density (PSAD), digital rectal examination findings, and prostate volume, individually and in combination, for the detection of prostate cancer (PCa) in biopsy-naive patients. We retrospectively analyzed 630 patients who underwent transrectal systematic prostate biopsy following prostate multiparametric magnetic resonance imaging. A standard 12-core biopsy procedure was performed. Univariate and multivariate analyses were performed to determine the significant predictors of clinically significant cancer but not PCa. The median age, PSA level, and PSAD were 70 years, 8.6 ng/mL, and 0.18 ng/mL/mL, respectively. A total of 374 (59.4%) of 630 patients were biopsy-positive for PCa, and 241 (64.4%) of 374 were diagnosed with clinically significant PCa (csPCa). The PI-RADS v2 score and PSAD were independent predictors of PCa and csPCa. The PI-RADS v2 score of 5 regardless of the PSAD value, or PI-RADS v2 score of 4 plus a PSAD of <0.3 ng/mL/mL, was associated with the highest csPCa detection rate (36.1%-82.1%). Instead, the PI-RADS v2 score of <3 and PSAD of <0.3 ng/mL/mL yielded the lowest risk of csPCa. The combination of the PI-RADS v2 score and PSAD could prove to be a helpful and reliable diagnostic tool before performing prostate biopsies. Patients with a PI-RADS v2 score of <3 and PSAD of <0.3 ng/mL/mL could potentially avoid a prostate biopsy.


Recombinant human platelet-derived growth factor enhances repair of cutaneous full-thickness excision by increasing the phosphorylation of extracellular signal-regulated kinase in diabetic rat.
To investigate the possible signaling mechanisms by which recombinant human platelet-derived growth factor (rhPDGF) accelerated healing of cutaneous wound in diabetic rats. Four full-thickness skin wounds were incised in the back of 26 male Wistar diabetic rats. The wounded rats were divided into 3 groups (7 or 8 rats each group). One group without treatment was used as a control, and the other 2 groups were treated with rhPDGF at a dose of 7.0 microg/cm2 wound or vehicle (DMSO/0.9% NaCl, vol/vol 1:1) from 1 to 14 days. The wound healing was evaluated by the measurements of the wound volume and area. Immunofluorescent and immunohistochemical staining were used to examine the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and the expression of proliferative cell nuclear antigen (PCNA), respectively. Granulation tissue appeared in the bed of wound after injury. The number of blood capillary buds and fibroblasts was greater in the rhPDGF-treated group than that in the other 2 groups. A lot of inflammatory cells infiltration and collagen deposition were observed in the wound. The wound-volume in the rhPDGF-treated group was smaller than that in control group (P < 0.05). The reepithelialization rate in rhPDGF-treated group was higher than that in the other 2 groups at 7 days after injury (P < 0.05). The expression of PCNA in reparative cells was higher in rhPDGF-treated group than in control group or vehicle-treated group at 3,7 days after injury (P < 0.05). The phosphorylation of ERK1/2 was stronger in rhPDGF-treated group than that in control group or vehicle group at 7 and 14 days after injury (P < 0.05). These results suggest that rhPDGF accelerates wound healing and improves healing quality by increasing the phosphorylation of ERK1/2.


Non-Targeted LC-MS/MS Assay for Screening Over 100 Lipid Mediators from ARA, EPA, and DHA in Biological Samples Based on Mass Spectral Fragmentations.
A non-targeted strategy to simultaneously screen for over 100 lipid mediators from ω-6 ARA and ω-3 EPA and DHA fatty acids is presented. The method based on an extensive study of fragmentation patterns obtained by SPE-LC-MS/MS analysis-provided fingerprints to comprehensively elucidate and identify lipid mediators in biological samples. Many of these metabolites are associated to metabolic disorders, inflammatory, immune and oxidative stress. The methodology consisted of a three-step procedure. (1) SPE extraction of compounds from plasma and adipose tissue was followed by LC-MS/MS analysis operating in full scan mode. The methodology was validated for a group of 65 metabolites using standards. SPE recoveries ranged from 29-134% and matrix effect from 10-580%. LOD and LOQ ranged from 0.01 to 1765 ng/mL and 0.03 to 5884 ng/mL respectively, similarly than current analytical strategies based on MRM mode. (2) An extensive study of the mass spectra of a wide range of compounds was done to stablish a specific fragmentation pattern. Interestingly, illustrative fragmentations and new specific transitions to identify EPA and DHA lipid mediators have been innovatively established. (3) After analysis, 30 lipid mediators were tentatively identified in plasma and 35 in adipose tissue of rats according to the pre stablished fragmentation patterns. The hypothetical identification of compounds was validated by using reference standards. Around 85-90% of proposed identifications were correctly assigned and only 4 and 3 identifications failed in adipose tissue and plasma, respectively. The method allowed the identification of these metabolites without losing information by the use of predefined ions list. Therefore, the use of full scan mode together with the study of fragmentation patterns provided a novel and stronger analytical tool to study the complete profile of lipid mediators in biological samples than the analysis through MRM based methods. Importantly, no analytical standards were required at this qualitative screening stage and the performance and sensitivity of the assay were very similar to that of a MRM method.


Sex Differences in Cardiac Troponin I and T and the Prediction of Cardiovascular Events in the General Population.
Cardiac troponin concentrations differ in women and men, but how this influences risk prediction and whether a sex-specific approach is required is unclear. We evaluated whether sex influences the predictive ability of cardiac troponin I and T for cardiovascular events in the general population. High-sensitivity cardiac troponin (hs-cTn) I and T were measured in the Generation Scotland Scottish Family Health Study of randomly selected volunteers drawn from the general population between 2006 and 2011. Cox-regression models evaluated associations between hs-cTnI and hs-cTnT and the primary outcome of cardiovascular death, myocardial infarction, or stroke. In 19 501 (58% women, mean age 47 years) participants, the primary outcome occurred in 2.7% (306/11 375) of women and 5.1% (411/8126) of men during the median follow-up period of 7.9 (IQR, 7.1-9.2) years. Cardiac troponin I and T concentrations were lower in women than men (P < 0.001 for both), and both were more strongly associated with cardiovascular events in women than men. For example, at a hs-cTnI concentration of 10 ng/L, the hazard ratio relative to the limit of blank was 9.7 (95% CI 7.6-12.4) and 5.6 (95% CI 4.7-6.6) for women and men, respectively. The hazard ratio for hs-cTnT at a concentration of 10 ng/L relative to the limit of blank was 3.7 (95% CI 3.1-4.3) and 2.2 (95% CI 2.0-2.5) for women and men, respectively. Cardiac troponin concentrations differ in women and men and are stronger predictors of cardiovascular events in women. Sex-specific approaches are required to provide equivalent risk prediction.


Functional role of the amino-terminal mobile segment in catalysis by porcine cytosolic aspartate aminotransferase. Critical importance of Val17 and Phe18 for productive binding of substrates.
A notable feature of porcine cytosolic aspartate aminotransferase is the closure of the active site cleft by a mobile amino-terminal segment (residues 15-40) upon binding substrate. The functional roles of Val17 and Phe18, residues that are part of the mobile loop, have been studied in the site-directed mutants in which the size and hydrophobic nature of these residues have been changed. Absorption, circular dichroism spectra, susceptibility to protease 401, and thermal stability did not differ appreciably between wild type and mutant enzymes. In the overall transamination between aspartate and 2-oxoglutarate, V17A represented a typical Km mutant while V17I retained the substrate binding affinity fairly well. In contrast, replacement of Phe18 by Ala resulted in a large decrease in both catalytic rate and binding affinity for substrates. F18W, F18Y, and F18H showed a moderate decrease in kcat and a considerable increase in Km values. Single-turnover reactions with four individual substrates yielded analogous results to those obtained for the overall reaction and, in addition, revealed that k/Kd values of mutants F18A and F18H were over 10 times lower for C5 substrates (glutamate and 2-oxoglutarate) than those for C4 substrates (aspartate and oxalacetate). All mutant enzymes showed variously increased Kd values for substrate analogs such as 2-methylaspartate, succinate, and glutarate. 1H NMR observations of F18H, in which His18 served as a built-in probe, were in accord with the behavior that would be expected from the conformational transition. We conclude that, although Val17 and Phe18 may not be essential for catalysis, the presence of a bulky residue of appropriate size at each position is critical for productive binding of substrate.


[Changes of chemical compositions and sulfur dioxide residues of Gastrodiae Rhizoma with different storage times].
To study the effect of different storage time on the chemical compositions and sulfur dioxide residues of sulfur-fumigated Gastrodiae Rhizoma (GR), and provide scientific basis for solving the quality and safety issues of sulfur-fumigated traditional Chinese medicinal materials. GR, sulfur-fumigated GR and its medicinal slices were stored under the same conditions, and then 8 active ingredients and sulfur dioxide residues were measured respectively. The results showed that the content of gastrodins in sulfur-fumigated GR and its medicinal slices was significantly lower than that in the non-fumigated GR. Moreover, the content of sulfur dioxide residue in sulfur-fumigated GR was significantly higher than that in its medicinal slices. That is to say, sulfur fumigation degree had significantly higher effect on GR quality as compared with its medicinal slices. During the whole storage time (8 months), the content of the eight chemical components in GR was not changed greatly in general. However, after the storage for 4 months, the content of 8 components and sulfur dioxide residues in all of GR samples were significantly changed. In particular, the content of sulfur dioxide residue in GR medicinal materials decreased up to 50% or more.


Mechanisms of retroviral integration and mutagenesis.
Gene transfer vectors derived from oncoretroviruses or lentiviruses are the most robust and reliable tools to stably integrate therapeutic transgenes in human cells for clinical applications. Integration of these vectors in the genome may, however, have undesired effects caused by insertional deregulation of gene expression at the transcriptional or post-transcriptional level. The occurrence of severe adverse events in several clinical trials involving the transplantation of stem cells genetically corrected with retroviral vectors showed that insertional mutagenesis is not just a theoretical event, and that retroviral transgenesis is associated with a finite risk of genotoxicity. In addressing these issues, the gene therapy community offered a spectacular example of how scientific knowledge and technology can be put to work to understand the causes of unpredicted side effects, design new vectors, and develop tools and models to predict their safety and efficacy. As an added benefit, these efforts brought new basic knowledge on virus-host interactions and on the biology and dynamics of human somatic stem cells. This review summarizes the current knowledge on the interactions between retroviruses and the human genome and addresses the impact of target site selection on the safety of retroviral vector-mediated gene therapy.


Nuclear PLCbeta(1) acts as a negative regulator of p45/NF-E2 expression levels in Friend erythroleukemia cells.
It is well established that phospholipase C (PLC) beta(1) plays a role in the nuclear compartment and is involved in the signalling pathway that controls the switching of the erythroleukemia cells programming from an undifferentiated to a differentiated state. Constitutive overexpression of nuclear PLCbeta(1) has been previously shown to inhibit Friend cells differentiation. For further characterization, we investigated the localization of PLCbeta(1)a and PLCbeta(1)b in Friend cells by fusing their cDNA to enhanced green fluorescent protein (GFP). To investigate the potential target of nuclear PLCbeta(1) in Friend differentiation, we studied the expression of p45/NF-E2 transcription factor, which is an enhancer binding protein for expression of the beta-globin gene and the expression of GATA proteins that are important for the survival and differentiation of erythroid cells. Our data suggest that the overexpression of PLCbeta(1) (both 1a and 1b) only in the nuclear compartment significantly reduces the expression of p45/NF-E2. The effect observed is attributable to the specific action of nuclear PLCbeta(1) signalling given that GATA-1 and GATA-3 are not affected at all. Here we show the existence of a unique target, i.e. the transcription factor p45/NF-E2, whose expression is specifically inhibited by the nuclear signalling evoked by PLCbeta(1) forms.


Constructing Doctoral Leadership Scholarly Role Boundaries Through Intraprofessional Nursing Education.
We conducted an intraprofessional education pilot in which students created DNP/PhD leadership parameters through naturalistic classroom experience. Agile, participatory, action research methodology framed a project spanning 3 semesters. Students' multiple ways of knowing influenced a dynamic cycle of noticing, interpreting, acting, and reflecting. Faculty revised course content "in the moment" to meet unique student needs. Researchers analyzed data using qualitative, interpretive descriptive approach. Four key themes interpreted from intraprofessional education included (1) persistent evolutionary progression toward doctoral collaboration, (2) accelerating uptake of research- into-practice continuum, (3) intentional convergent/divergent curriculum, and (4) mutuality of leadership parameters.


Asymmetric hypsarrhythmia and infantile spasms in west syndrome.
The video encephalograms (EEGs) of 77 consecutive infantile spasms patients were evaluated for the presence of focal or asymmetric hypsarrhythmia and infantile spasms, to determine whether these findings were useful in predicting the presence of focal structural brain disease and were of any additional diagnostic or prognostic significance. Of the 77 patients with infantile spasms, 38% had focal or lateralized features present on video-EEG studies. Unilateral hypsarrhythmia and asymmetric ictal EEG changes during infantile spasms often occurred together: each always indicated the side of a focal or asymmetric structural cerebral lesion that was visible on computed tomographic or magnetic resonance imaging brain scan and was usually large. Clinically asymmetric infantile spasms were less common, always occurred in the presence of asymmetric ictal EEG changes, and did not appear to have additional localizing value. Lateralized hypsarrhythmia, with or without asymmetric infantile spasms, occurred in the presence of bilateral structural lesions that were more abnormal in the area of the greater EEG abnormality. Partial seizures also indicated symptomatic etiologies but were less localizing to visible focal lesions. Patients with symmetric hypsarrhythmia and infantile spasms rarely had focal/lateralized lesions visible on imaging studies. Although the majority of the symmetric group had structural brain disease, these brain lesions were diffuse, not lateralized. This group also included all patients who had cryptogenic etiology and normal development.


[Insult - Stroke].
A stroke is usually caused by ischaemia, less frequently by an intracerebral hemorrhage. The distinction between hemorrhagic and ischaemic strokes, as well as the identification of patients requiring immediate recanalization therapy, cannot be made by clinical examination but requires CT imaging. In most patients with an ischemic stroke, large parts of the affected brain area are functionless throughout the first hours, but not yet irreversibly damaged (penumbra tissue). These areas can be saved through early vessel recanalization. In the acute phase of ischaemic stroke, a delay of 15 minutes results in a significant worsening of the neurologic prognosis.With modern endovascular techniques, even patients with the most severe ischemic strokes have a fair chance of neurological recovery, if they are admitted to a specialised centre within the first few hours.


Suppression of Phagocytic Activity Leads to the Efficient Surface Modification of Macrophages with Liposomes for Developing a Biomimetic Drug Delivery System.
Macrophages selectively infiltrate the lesion sites of several diseases, including cancers, and, thus, have attracted attention as a biomimetic drug delivery carrier. To achieve the efficient drug loading of macrophages with minimal cytotoxicity, drugs are preferably encapsulated into nanoparticles, such as liposomes, and modified on the surface of macrophages rather than being incorporated into cells. However, liposomes are rapidly taken up by macrophages after binding to the cell surface because of their strong phagocytic activity. To overcome this, we herein attempted to modify the surface of macrophages with liposomes by suppressing their phagocytic activity using a pretreatment with anionic liposomes. We confirmed that 1,2-distearoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DSPG)- and cholesterol-rich anionic liposomes were efficiently taken up by RAW264.7 murine macrophage-like cells. Furthermore, the cellular uptake of anionic liposomes by RAW264.7 cells was higher in the absence of fetal bovine serum (FBS) than in its presence. Moreover, the viability of RAW264.7 cells was maintained above 90% when cells were incubated with anionic liposomes for 3 h, whereas viability was markedly decreased after a 24-h incubation. Based on these results, we pretreated RAW264.7 cells by an incubation with DSPG- and cholesterol-rich liposomes for 3 h in the absence of FBS. This pretreatment significantly inhibited the internalization of other liposomes, which subsequently bound to the cell surface. Therefore, we succeeded in modifying the surface of macrophages with liposomes, and liposome-modified macrophages have potential as a biomimetic active drug delivery carrier.


The liver isoform of carnitine palmitoyltransferase I is activated in neonatal rat cardiac myocytes by hypoxia.
Fatty acids are the preferred substrate of ischemic, reperfused myocardium and may account for the decreased cardiac efficiency during aerobic recovery. Neonatal cardiac myocytes in culture respond to hypoxia/serum- and glucose-free medium by a slow decline in ATP which reverses upon oxygenation. This model was employed to examine whether carnitine palmitoyltransferase I (CPT-I) modulates high rates of beta-oxidation following oxygen deprivation. After 5 h of hypoxia, ATP levels decline to 30% control values and CPT-I activity is significantly stimulated in hypoxic myocytes with no alteration in cellular carnitine content or in the release of the mitochondrial matrix marker, citrate synthase. This stimulation was attributed to an increase in the affinity of hypoxic CPT-I for carnitine, suggesting that the liver CPT-I isoform is more dominant following hypoxia. However, there was no alteration in hypoxic CPT-I inhibition by malonyl-CoA. DNP-etomoxiryl-CoA, a specific inhibitor of the liver CPT-I isoform, uncovered identical Michaelis kinetics of the muscle isoform in control and hypoxic myocytes with activation of the liver isoform. Northern blotting did not reveal any change in the relative abundance of mRNA for the liver vs. the muscle CPT-I isoforms. The tyrosine phosphatase inhibitor, pervanadate, reversed the hypoxia-induced activation of CPT-I and returned the affinity of cardiac CPT-I for carnitine to control. Reoxygenation was also associated with a return of CPT-I activity to control levels. The data demonstrate that CPT-I is activated upon ATP depletion. Lower enzyme activities are present in control and reoxygenated cells where ATP is abundant or when phosphatases are inhibited. This is the first suggestion that phosphorylation may modulate the activity of the liver CPT-I isoform in heart.


Dissipative soliton stabilization by several nonlinear gradient terms.
We study a single cubic complex Ginzburg-Landau equation with nonlinear gradient terms analytically and numerically. This single equation allows for the existence of stable dissipative solitons exclusively due to nonlinear gradient terms. We shed new light on the feedback loop, leading to dissipative solitons (DSs) by analyzing a mechanical analog as a function of the magnitude of the amplitude. In addition, we present analytic results incorporating four nonlinear gradient terms and derive necessary conditions for the existence of DSs. We also elucidate in detail for the case of the Raman contribution the scaling behavior for the limit of the vanishing Raman term.


Trends in bariatric surgical procedures.
The increasing prevalence and associated sociodemographic disparities of morbid obesity are serious public health concerns. Bariatric surgical procedures provide greater and more durable weight reduction than behavioral and pharmacological interventions for morbid obesity. To examine trends for elective bariatric surgical procedures, patient characteristics, and in-hospital complications from 1998 to 2003 in the United States. The Nationwide Inpatient Sample was used to identify bariatric surgery admissions from 1998-2002 (with preliminary data for 12 states from 2003) using International Classification of Diseases, Ninth Revision, codes for foregut surgery with a confirmatory diagnosis of obesity or by diagnosis related group code for obesity surgery. Annual estimates and trends were determined for procedures, patient characteristics, and adjusted complication rates. Trends in bariatric surgical procedures, patient characteristics, and complications. The estimated number of bariatric surgical procedures increased from 13,365 in 1998 to 72,177 in 2002 (P<.001). Based on preliminary state-level data (1998-2003), the number of bariatric surgical procedures is projected to be 102 794 in 2003. Gastric bypass procedures accounted for more than 80% of all bariatric surgical procedures. From 1998 to 2002, there were upward trends in the proportion of females (81% to 84%; P = .003), privately insured patients (75% to 83%; P = .001), patients from ZIP code areas with highest annual household income (32% to 60%, P<.001), and patients aged 50 to 64 years (15% to 24%; P<.001). Length of stay decreased from 4.5 days in 1998 to 3.3 days in 2002 (P<.001). The adjusted in-hospital mortality rate ranged from 0.1% to 0.2%. The rates of unexpected reoperations for surgical complications ranged from 6% to 9% and pulmonary complications ranged from 4% to 7%. Rates of other in-hospital complications were low. These findings suggest that use of bariatric surgical procedures increased substantially from 1998 to 2003, while rates of in-hospital complications were stable and length of stay decreased. However, disparities in the use of these procedures, with disproportionate and increasing use among women, those with private insurance, and those in wealthier ZIP code areas should be explored further.


A Young Boy with 21q21.1 Microdeletion Showing Speech Delay, Spastic Diplegia, and MRI Abnormalities: Original Case Report.
Chromosome 21q deletion syndrome is a rare disorder affecting the long arm of chromosome 21 and manifesting with wide phenotypic features depending on the size and position of the deleted region. In the syndrome, three distinct deleted regions have been distinguished: region 1, from the centromere to approximately 31.2 Mb (21q11.2-q22.11); region 2, from 31.2 to 36 Mb (21q22.11-q22.12); and region 3, from 36 to 37.5 Mb to the telomere (21q22.12-q22.3). The clinical features are highly variable manifesting with mild, poorly recognizable signs or with severe symptoms including craniofacial dysmorphism, growth failure, developmental delay, behavioral/affective abnormalities, and systemic malformations. We report here the case of a young boy with speech delay, mild spastic diplegia, and brain anomalies on magnetic resonance imaging (MRI). The genetic analysis displayed a microdeletion of the long arm of chromosome 21 approximately extending up to 1.08 Mb. Clinical presentation of the patient and cases of 21q21 deletion reported by the literature are discussed.


Nonadiabatic dynamics near metal surfaces under Floquet engineering: Floquet electronic friction vs Floquet surface hopping.
In the previous study Wang and Dou [J. Chem. Phys. 158, 224109 (2023)], we have derived a Floquet classical master equation (FCME) to treat nonadiabatic dynamics near metal surfaces under Floquet engineering. We have also proposed a trajectory surface hopping algorithm to solve the FCME. In this study, we map the FCME into a Floquet Fokker-Planck equation in the limit of fast Floquet driving and fast electron motion as compared to nuclear motion. The Fokker-Planck equation is then being solved using Langevin dynamics with explicit friction and random force from the nonadiabatic effects of hybridized electrons and Floquet states. We benchmark the Floquet electronic friction dynamics against Floquet quantum master equation and Floquet surface hopping. We find that Floquet driving results in a violation of the second fluctuation-dissipation theorem, which further gives rise to heating effects.


Solvent-dependent segmental dynamics in intrinsically disordered proteins.
Protein and water dynamics have a synergistic relationship, which is particularly important for intrinsically disordered proteins (IDPs), although the details of this coupling remain poorly understood. Here, we combine temperature-dependent molecular dynamics simulations using different water models with extensive nuclear magnetic resonance (NMR) relaxation to examine the importance of distinct modes of solvent and solute motion for the accurate reproduction of site-specific dynamics in IDPs. We find that water dynamics play a key role in motional processes internal to "segments" of IDPs, stretches of primary sequence that share dynamic properties and behave as discrete dynamic units. We identify a relationship between the time scales of intrasegment dynamics and the lifetime of hydrogen bonds in bulk water. Correct description of these motions is essential for accurate reproduction of protein relaxation. Our findings open important perspectives for understanding the role of hydration water on the behavior and function of IDPs in solution.


Improvements of growth, appetite, and physical activity in helminth-infected schoolboys 6 months after single dose of albendazole.
The effect of treatment for helminth infections on growth, appetite, and physical activity was investigated in Indonesian schoolchildren with Ascaris and Trichuris infections. Groups of schoolboys were selected for this substudy from a large study in which two groups received a single dose of 400 mg albendazole (AL, n = 86) and one group received an identical placebo (PL, n = 43). All boys were measured for parasitic infection, growth, appetite, and physical activity at baseline and 6 months after treatment. At baseline, all variables measured were not significantly different. After 6 months of treatment, the prevalence of Ascaris and Trichuris infections did not change significantly for both groups but the intensity of Ascaris and Trichuris infections significantly reduced in both groups (p<0.05) except for Trichuris in the PL group. Increases in mid-arm circumference and height-for-age, after treatment, in the AL group were significantly greater than in the PL group (p<0.05). In addition, appetite scores were higher in the AL group than in the PL group (P = 0.014). Free play activity, measured by Caltrac accelerometers, increased by 28% in the AL group after treatment (P = 0.004) and did not change in the PL group. We conclude that treatment with a single dose of albendazole may improve growth, appetite, and activity in areas with a high transmission of helminth infections.


A comprehensive compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects.
We have expanded a former compartmental model of blood glucose regulation for healthy and type 2 diabetic subjects. The former model was a detailed physiological model which considered the interactions of three substances, glucose, insulin and glucagon on regulating the blood sugar. The main drawback of the former model was its restriction on the route of glucose entrance to the body which was limited to the intravenous glucose injection. To handle the oral glucose intake, we have added a model of glucose absorption in the gastrointestinal tract to the former model to address the resultant variations of blood glucose concentrations following an oral glucose intake. Another model representing the incretins production in the gastrointestinal tract along with their hormonal effects on boosting pancreatic insulin production is also added to the former model. We have used two sets of clinical data obtained during oral glucose tolerance test and isoglycemic intravenous glucose infusion test from both type 2 diabetic and healthy subjects to estimate the model parameters and to validate the model results. The estimation of model parameters is accomplished through solving a nonlinear optimization problem. The results show acceptable precision of the estimated model parameters and demonstrate the capability of the model in accurate prediction of the body response during the clinical studies.


[Epidemiologic study of injuries in infants].
Eight hundred fifty-four injuries to children under 2 years of age were analysed. Injuries were more frequent in the second year than in younger infants. No major male predominance was observed. Most injuries were related to home accidents and mainly to mechanical trauma. Burns and poisonings were less frequent but their percentage was higher during the second year. Most lesions were benign, except in case of fall from a height (window falls, falls from baby furniture). Medical cost was rather high due to a 17% rate of hospital admissions and a 4 days average length of stay. Two fatal cases were registered. Sequelae appeared to be rare. Child abuse was seldom suspected and reason for that remains unclear. The study of accidents mechanism leads to think that teaching young children the control of hazardous situations should play a large part in prevention.


Erythema nodosum and sarcoid granulomas - letting the cat out of the bag.
A 41-year-old woman presented with a violaceous papule on the dorsum of the hand, large ipsilateral axillary lymphadenopathy, and tender, erythematous, subcutaneous nodules on the legs. Accompanying signs included fever, ankle swelling, and bilateral red eye. She recalled having a previous exposure to kittens one month before and had a positive family history for sarcoidosis. Histological examination of the hand lesion showed sarcoidal granulomas with positive Bartonella henselae DNA, whereas a biopsy done on the leg nodules was compatible with erythema nodosum. Cat scratch disease (CSD) typically presents as a tender regional lymphadenopathy preceded by an inoculation papule with spontaneous resolution occurring between 8-16 weeks. Cutaneous manifestations of CSD are rare, with erythema nodosum accompanying only 0.6% of cases. Although speculative, the background of a positive family history for sarcoidosis may explain the atypical presentation of this case, with red eye, persistent arthralgia, and associated sarcoidal granulomas.


MR imaging of the pancreas.
Magnetic resonance (MR) imaging sequences that reduce image artifact in upper abdominal examinations herald a role for MR in the investigation of pancreatic disease. Sequences that have been successful in imaging the pancreas are T1-weighted breath-hold gradient-echo imaging, which avoids phase artifact caused by respiration, and T1-weighted fat-suppressed spin-echo imaging, which reduces breathing artifact, removes chemical shift artifact, and improves dynamic range of signal intensities. Breath-hold gradient-echo sequences provide useful diagnostic information when used prior to and immediately following bolus intravenous administration of gadopentetate dimeglumine. This review addresses current clinically important roles for MR imaging and emphasizes the detection of islet cell tumors, non-organ-deforming pancreatic ductal adenocarcinomas, and the distinction between chronic pancreatitis and cancer.


Aryl hydrocarbon receptor-dependent suppression by 2,3,7, 8-tetrachlorodibenzo-p-dioxin of IgM secretion in activated B cells.
The immune system has been identified as a sensitive target for the toxic effects produced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Furthermore, the B cell has been identified as a sensitive cellular target of TCDD by previous cell-type fractionation studies from this laboratory. The mechanism responsible for the immunotoxic effects produced by TCDD is unclear; however, many of the biological effects of TCDD are thought to be mediated by the aryl hydrocarbon receptor (AhR). Here, we describe two B cell lines that differ considerably in their expression of the AhR and in their sensitivity to TCDD. Our results demonstrated a marked expression of the AhR protein in the CH12.LX B cell line but not in the BCL-1 B cell line. Transcripts for the AhR were not detected by reverse transcriptase-polymerase chain reaction in the BCL-1 cells. The AhR nuclear translocator (ARNT) protein was highly expressed in both cell lines. In addition, the AhR and ARNT are functional in CH12.LX cells as demonstrated by TCDD-induced CYP1A1 induction. TCDD did not induce CYP1A1 in BCL-1 cells. Furthermore, TCDD treatment resulted in suppression of lipopolysaccharide (LPS)-induced IgM secretion in CH12.LX cells. Conversely, TCDD-induced inhibition of IgM secretion was not demonstrated in LPS-stimulated BCL-1 cells, implicating a role for the AhR in the inhibition of B cell effector function. LPS-induced differentiation of the CH12.LX cells also resulted in a marked induction of Ahr expression which was not induced in LPS-stimulated BCL-1 cells. These studies have implicated the AhR as a critical factor in TCDD-induced inhibition of IgM secretion and have demonstrated an induction of AhR gene and protein expression after B cell activation.


Cardiac tamponade following liver transplantation after intrapericardial control of the suprahepatic vena cava.
Transabdominal intrapericardial control of the suprahepatic inferior vena cava (SIVC) is a rather uncommon procedure occasionally required in conjunction with complicated liver transplantation (LT) and hepatobiliary surgery. Experience with this technique is limited. Here we report 6 cases of LT in which transabdominal intrapericardial control of the SIVC was necessary. After institutional review board approval was obtained, a single-center, retrospective review was conducted from January 1991 to December 2013 to identify adult cases (age > 18 years) of LT in which transabdominal intrapericardial isolation of the SIVC was necessary. Among 4102 adult LT cases in the study period, 6 such cases were identified. To gain access to the pericardial space, a 6- to 9-cm vertical incision was made above the SIVC. After reperfusion, the diaphragmatic incision was partially closed and selectively drained. Pericardial tamponade developed in 1 patient, and it necessitated emergent reoperation and widespread drainage. In conclusion, transabdominal intrapericardial isolation of the SIVC is easily achieved without the need for a separate thoracic incision. However, to be effective, the pericardial incision should be only partially closed, and the pericardial sac should be drained liberally. Such patients should be carefully monitored for signs and symptoms of pericardial tamponade, the development of which should prompt an immediate return to the operating room for emergent decompression and widespread drainage.


A New Deep Learning-based Dynamic Paradigm Towards Open-World Plant Disease Detection.
Plant disease detection has made significant strides thanks to the emergence of deep learning. However, existing methods have been limited to closed-set and static learning settings, where models are trained using a specific dataset. This confinement restricts the model's adaptability when encountering samples from unseen disease categories. Additionally, there is a challenge of knowledge degradation for these static learning settings, as the acquisition of new knowledge tends to overwrite the old when learning new categories. To overcome these limitations, this study introduces a novel paradigm for plant disease detection called open-world setting. Our approach can infer disease categories that have never been seen during the model training phase and gradually learn these unseen diseases through dynamic knowledge updates in the next training phase. Specifically, we utilize a well-trained unknown-aware region proposal network to generate pseudo-labels for unknown diseases during training and employ a class-agnostic classifier to enhance the recall rate for unknown diseases. Besides, we employ a sample replay strategy to maintain recognition ability for previously learned classes. Extensive experimental evaluation and ablation studies investigate the efficacy of our method in detecting old and unknown classes. Remarkably, our method demonstrates robust generalization ability even in cross-species disease detection experiments. Overall, this open-world and dynamically updated detection method shows promising potential to become the future paradigm for plant disease detection. We discuss open issues including classification and localization, and propose promising approaches to address them. We encourage further research in the community to tackle the crucial challenges in open-world plant disease detection. The code will be released at https://github.com/JiuqingDong/OWPDD.


Effect of infrared analyzer homogenization efficiency on repeatability of uncorrected fat A and fat B signals.
Poor repeatability by infrared milk analyzers may be caused by inefficient homogenization as a result of light scattering and the Christiansen effect. The objectives of this study were to identify instruments with good and poor homogenization efficiency and to determine if a difference exists in repeatability performance between instruments with good vs poor homogenization efficiency. Unhomogenized and homogenized portions of the same milk were tested 20 times consecutively on 22 instruments. An instrument was considered to have poor homogenization efficiency if the mean difference in the uncorrected signal between unhomogenized and homogenized portions of the same milk was > or = 1.43% of the fat test (i.e., > or = 0.05% at 3.5% fat). Instruments were evaluated for repeatability by calculating the sample standard deviation and the range of the latter 19 uncorrected readings for unhomogenized and homogenized milks. When repeatability was evaluated as a function of homogenization efficiency, there was a significant (p = 0.001) correlation between poor homogenization efficiency and poor repeatability when testing unhomogenized milk but not when testing homogenized milk. Improved homogenizer performance within infrared milk analyzers is needed to improve the repeatability of raw milk testing.


A panel of real-time PCR assays for the detection of Bourbon virus, Heartland virus, West Nile virus, and Trypanosoma cruzi in major disease-transmitting vectors.
Vector-borne pathogens, such as Bourbon virus (BRBV), Heartland virus (HRTV), West Nile virus (WNV), and Trypanosoma cruzi (TCZ) are a great threat to public health and animal health. We developed a panel of TaqMan real-time PCR assays for pathogen surveillance. PCR targets were selected based on nucleic acid sequences deposited in GenBank. Primers and probes were either designed de novo or selected from publications. The coverages and specificities of the primers and probes were extensively evaluated by performing BLAST searches. Synthetic DNA or RNA fragments (gBlocks) were used as PCR templates in initial assay development and PCR positive controls in subsequent assay validation. For operational efficiency, the same thermocycling profile was used in BRBV, HRTV, and WNV reverse-transcription quantitative PCR (RT-qPCR) assays, and a similar thermocycling profile without the initial reverse-transcription step was used in TCZ qPCR. The assays were optimized by titrating primer and probe concentrations. The analytical sensitivities were 100, 100, 10, and 10 copies of gBlock per reaction for BRBV (Cq = 36.0 ± 0.7), HRTV (Cq = 36.6 ± 0.9), WNV (Cq = 35.5 ± 0.4), and TCZ (Cq = 38.8 ± 0.3), respectively. PCR sensitivities for vector genomic DNA or RNA spiked with gBlock reached 100, 100, 10, and 10 copies per reaction for BRBV, HRTV, WNV, and TCZ, respectively. PCR specificity evaluated against a panel of non-target pathogens showed no significant cross-reactivity. Our BRBV, HRTV, WNV, and TCZ PCR panel could support epidemiologic studies and pathogen surveillance.


Potential Impact of Fluopyram on the Frequency of the D123E Mutation in Alternaria solani.
Succinate dehydrogenase-inhibiting (SDHI) fungicides have been widely applied in commercial potato (Solanum tuberosum L.) fields for the control of early blight, caused by Alternaria solani Sorauer. Five-point mutations on three AsSdh genes in A. solani have been identified as conferring resistance to SDHI fungicides. Recent work in our laboratory determined that A. solani isolates possessing the D123E mutation, or the substitution of aspartic acid for glutamic acid at position 123 in the AsSdhD gene, were collected at successively higher frequencies throughout a 3-year survey. In total, 118 A. solani isolates previously characterized as possessing the D123E mutation were evaluated in vitro for boscalid and fluopyram sensitivity. Over 80% of A. solani isolates with the D123E mutation evaluated were determined to be highly resistant to boscalid in vitro. However, effective concentration at which the fungal growth is inhibited by 50% values of isolates with the D123E mutation to fluopyram, ranging from 0.2 to 3 µg/ml, were sensitive and only slightly higher than those of baseline isolates to fluopyram, which ranged from 0.1 to 0.6 µg/ml. Five A. solani isolates with the D123E mutation were further evaluated in vivo for percent disease control obtained from boscalid and fluopyram compared with two wild-type isolates, three isolates possessing the F129L mutation, two isolates possessing the H134R mutation, two isolates possessing the H133R mutation, and one isolate with the H278R mutation. Relative area under the dose response curve values for boscalid and fluopyram were significantly lower for all five D123E-mutant isolates, demonstrating reduced disease control in vivo. In field trials, the frequency of A. solani isolates with the D123E mutation recovered from treatments receiving an in-furrow application of fluopyram ranged from 5 to 37%, which was significantly higher compared with treatments receiving foliar applications of standard protectants, in which the frequency of the D123E mutation in isolates ranged from 0 to 2.5%. Results suggest that A. solani isolates possessing the D123E mutation have a selective advantage under the application of fluopyram compared with SDHI-sensitive isolates, as well as isolates possessing other mutations conferring SDHI resistance. These data illustrate the importance of implementing fungicide resistance management strategies and cautions the use of fluopyram for in-furrow applications that target other pathogens of potato.


[Military medicine from the aspect of medical informatics].
The aggression and the war that has been imposed on Bosnia and Herzegovina is an excellent confirmation for the necessity of information as a resource of today. To be more specific, the information became a resource more important than the material or the energy, since it was exactly through the freedom of the information system and communications that the aggressor has managed to maximally engage and use all the other resources, and to considerably block our own and put it out of function. The health system, as the most important social system after the Defence Forces and the state authorities, had to create its own information system immediately at the beginning of the war and put it into function of health protection of the population in general, and at the disposal of the BH Army soldiers. On the other hand the indicators of the aggression level and all of its negative attributes (killed soldiers, murdered or injured civilians, mode, character and the level of brutality during injuring the victims and the like) are significantly repercussioned towards ours, the aggressors' and the foreign media and the structures from which depends further outcome of the war. And, finally, all the actions within the system of health activities, from offering healts protection, through the application of methods, techniques, technologies, resources, medicaments etc, also have its educative quality and value, furthermore, since the war which still lasts on these grounds, according to its character and specificity, has been so far unregistered in human history of the world.(ABSTRACT TRUNCATED AT 250 WORDS)


Mechanical Ventilation and Extracorporeal Membrane Oxygenation as a Bridging Strategy to Lung Transplantation: Significant Gains in Survival.
Mechanical ventilation (MV) and extracorporeal membrane oxygenation (ECMO) are increasingly used to bridge patients to lung transplantation. We investigated the impact of using MV, with or without ECMO, before lung transplantation on survival after transplantation by performing a retrospective analysis of 826 patients who underwent transplantation at our high-volume center. Recipient characteristics and posttransplant outcomes were analyzed. Most lung transplant recipients (729 patients) did not require bridging; 194 of these patients were propensity matched with patients who were bridged using MV alone (48 patients) or MV and ECMO (49 patients). There was no difference in overall survival between the MV and MV+ECMO groups (p = 0.07). The MV+ECMO group had significantly higher survival conditioned on surviving to 1 year (median 1,811 days ([MV] vs. not reached ([MV+ECMO], p = 0.01). Recipients in the MV+ECMO group, however, were more likely to require ECMO after lung transplantation (16.7% MV vs. 57.1% MV+ECMO, p < 0.001). There were no differences in duration of postoperative MV, hospital stay, graft survival, or the incidence of acute rejection, renal failure, bleeding requiring reoperation, or airway complications. In this contemporary series, the combination of MV and ECMO was a viable bridging strategy to lung transplantation that led to acceptable patient outcomes.


Development of a highly sensitive probe for measuring oxygen in biological tissues.
The development of a new highly sensitive electron paramagnetic resonance probe suitable for measuring low oxygen concentrations in biological cells and tissues is reported. This probe consists of a char prepared from glucose. This char exhibits an EPR spectrum with marked oxygen-dependent linewidth broadening with a linewidth of 0.145 G under anaerobic conditions and a marked oxygen-dependent linewidth broadening of 14.5 G per percent oxygen. This marked oxygen-dependent broadening allows oxygen tensions to be measured down to mTorr values which was not previously possible with any other technique. This oximetry probe was applied to perform measurements to determine the severity of hypoxia in the ischemic rat heart. It was observed that after 10 minutes of ischemia, the oxygen concentration was 440 mTorr and it continued to fall over a 70 minute period to a value of 120 mTorr. Thus, absolute anoxia did not occur in the ischemic heart. The glucose char oximetry probe enables the measurement of oxygen in hypoxic cells and tissues with higher sensitivity than previously possible and therefore it can serve as a new important tool in characterizing the effects of low oxygen tensions in metabolic control and cellular injury.


Gastric reflux measurements during duodenal infusion with saline, acid and fat.
Duodeno-gastric reflux measured in 18 patients without gastrointestinal complaints showed a wide variation of regurgitation rates. Duodeno-gastric reflux was significantly diminished during duodenal acidification. The effect was more intense and lasted longer after olive oil infused intraduodenally, leading practically to a closed pylorus.


Ultra-thick inkjet-printed quantum dots layer for full-color micro-LED displays.
Micro-LEDs have promising development potential in display applications because of their outstanding performance. Achieving a full-color display based on micro-LEDs is one of the most important issues in commercial applications. In this paper, an effective method based on quantum dots and blue micro-LEDs was developed. Using an etching method, a thick black matrix was fabricated to reduce crosstalk and form a thick bank for quantum dots. Quantum dots were deposited in a thick black matrix using inkjet printing technology. With blue micro-LEDs, inkjet-printed quantum dot films can realize effective color conversion. The integrated blue micro-LEDs and red/green quantum dot films can achieve full-color displays without color filters, because the blue light leakage in the color conversion film can be reduced by the quantum dots themselves. The results suggest that inkjet-printed quantum dots are a promising way to achieve full-color micro-LED displays.


Oscillations in a patchy environment disease model.
For a single patch SIRS model with a period of immunity of fixed length, recruitment-death demographics, disease related deaths and mass action incidence, the basic reproduction number R(0) is identified. It is shown that the disease-free equilibrium is globally asymptotically stable if R(0)<1. For R(0)>1, local stability of the endemic equilibrium and Hopf bifurcation analysis about this equilibrium are carried out. Moreover, a practical numerical approach to locate the bifurcation values for a characteristic equation with delay-dependent coefficients is provided. For a two patch SIRS model with travel, it is shown that there are several threshold quantities determining its dynamic behavior and that travel can reduce oscillations in both patches; travel may enhance oscillations in both patches; or travel can switch oscillations from one patch to another.