Functional heterogeneity of human CD34(+) cells isolated in subcompartments of the G0 /G1 phase of the cell cycle.
Using simultaneous Hoechst 33342 (Hst) and Pyronin Y (PY) staining for determination of DNA and RNA content, respectively, human CD34(+) cells were isolated in subcompartments of the G0 /G1 phase of the cell cycle by flow cytometric cell sorting. In both bone marrow (BM) and mobilized peripheral blood (MPB) CD34(+) cells, primitive long-term hematopoietic culture-initiating cell (LTHC-IC) activity was higher in CD34(+) cells isolated in G0 (G0CD34(+) cells) than in those residing in G1 (G1CD34(+) cells). However, as MPB CD34(+) cells displayed a more homogeneous cell-cycle status within the G0 /G1 phase and a relative absence of cells in late G1 , DNA/RNA fractionation was less effective in segregating LTHC-IC in MPB than in BM. BM CD34(+) cells belonging to four subcompartments of increasing RNA content within the G0 /G1 phase were evaluated in functional assays. The persistence of CD34 expression in suspension culture was inversely correlated with the initial RNA content of test cells. Multipotential progenitors were present in G0 or early G1 subcompartments, while lineage-restricted granulomonocytic progenitors were more abundant in late G1 . In vitro hematopoiesis was maintained for up to 6 weeks with G0CD34(+) cells, whereas production of clonogenic progenitors was more limited in cultures initiated with G1CD34(+) cells. To test the hypothesis that primitive LTHC-ICs would reenter a state of relative quiescence after in vitro division, BM CD34(+) cells proliferating in ex vivo cultures were identified from their quiescent counterparts by a relative loss of membrane intercalating dye PKH2, and were further fractionated with Hst and PY. The same functional hierarchy was documented within the PKH2(dim) population whereby LTHC-IC frequency was higher for CD34(+) cells reselected in G0 after in vitro division than for CD34(+) cells reisolated in G1 or in S/G2 + M. However, the highest LTHC-IC frequency was found in quiescent PKH2(bright) CD34(+) cells. Together, these results support the concept that cells with distinct hematopoietic capabilities follow different pathways during the G0 /G1 phase of the cell cycle both in vivo and during ex vivo culture.


Amplified oligonucleotide sensing in microliter volumes containing copper ions by solution streaming.
We present a simple, cost-effective design for amplifying oligodeoxynucleotide (ODN) sensing, in microliter ODN volumes containing copper ions, by solution streaming (bubbling). The inert gas streaming (bubbling) at a constant pressure of 0.04 bar drives the motion of a 30-microL ODN droplet containing a three-electrode circuit (inverted drop microcell), and in the presence of copper ions offers an approximately 50-times improvement in the detection of ODN samples. The detection of ODNs at the carbon paste electrode is based on the enhancement of the oxidation peaks of purine bases (adenine and guanine) by the anodic stripping of the electrochemically accumulated complex of Cu(I) with purine base residues of acid hydrolyzed ODN samples (Cu(I)-ahODN complex). We used the proposed method for (i) the determination of the percentage content of adenine and guanine units within analyzed ODN samples at subnanomolar concentrations (related to monomer content) and (ii) the detection of the (TTC)n triplet expansion using magnetic DNA hybridization with reporter probes containing guanine units (the TTC trinucleotide repeat expansion is associated with serious hereditary diseases, including Friedreich ataxia).


Coaching Caregivers of Children who are Deaf or Hard of Hearing.
This study explored the impact of a caregiver training intervention on caregivers' use of individually selected facilitative language techniques (FLTs) with their children (ranging from 14 to 27 months of age) who were deaf or hard of hearing. In addition, the study sought to identify real-time relationships between caregiver input and child output. Finally, the Language ENvironment Analysis (LENA) system was used to assess generalization of intervention gains to the natural environment. Intervention was delivered via a telehealth approach to train four caregivers to utilize a targeted FLT with their child. Pre-baseline LENA recordings guided selection of treatment targets, and post-intervention LENA recordings were used to assess generalization to the natural environment. All caregivers demonstrated increases in use of their targeted FLT in structured intervention sessions and maintained them 2-6 weeks after the conclusion of intervention. All children demonstrated an increase in their developmentally appropriate vocalizations in tandem with caregiver FLT increases. Results indicate that coaching can be used to increase caregiver responsiveness, changes which were found to carry over into the home environment.


Prediction of rupture risk in abdominal aortic aneurysm during observation: wall stress versus diameter.
We previously showed that peak abdominal aortic aneurysm (AAA) wall stress calculated for aneurysms in vivo is higher at rupture than at elective repair. The purpose of this study was to analyze rupture risk over time in patients under observation. Computed tomography (CT) scans were analyzed for patients with AAA when observation was planned for at least 6 months. AAA wall stress distribution was computationally determined in vivo with CT data, three-dimensional computer modeling, finite element analysis (nonlinear hyperelastic model depicting aneurysm wall behavior), and blood pressure during observation. Analysis included 103 patients and 159 CT scans (mean follow-up, 14 +/- 2 months per CT). Forty-two patients were observed with no intervention for at least 1 year (mean follow-up, 28 +/- 3 months). Elective repair was performed within 1 year in 39 patients, and emergent repair was performed in 22 patients (mean, 6 +/- 1 month after CT) for rupture (n = 14) or acute severe pain. Significant differences were found for initial diameter (observation, 4.9 +/-.1 cm; elective repair, 5.9 +/-.1 cm; emergent repair, 6.1 +/-.2 cm; P <.0001) and initial peak wall stress (38 +/- 1 N/cm(2), 42 +/- 2 n/cm(2), 58 +/- 4 N/cm(2), respectively; P <.0001), but peak wall stress appeared to better differentiate patients who later required emergent repair (elective vs emergent repair: diameter, 3% difference, P =.5; stress, 38% difference, P <.0001). Receiver operating characteristic (ROC) curves for predicting rupture were better for peak wall stress (sensitivity, 94%; specificity,81%; accuracy, 85% [with 44 N/cm(2) threshold]) than for diameter (81%, 70%, 73%, respectively [with optimal 5.5 cm threshold). With proportional hazards analysis, peak wall stress (relative risk, 25x) and gender (relative risk, 3x) were the only significant independent predictors of rupture. For AAAs under observation, peak AAA wall stress seems superior to diameter in differentiating patients who will experience catastrophic outcome. Elevated wall stress associated with rupture is not simply an acute event near the time of rupture.


Health of children in Jamaica: the new health realities.
The World Health Organization developed a mathematical technique, which discount the life expectancy by the years spent in disability or morbidity. Health, therefore, must be more than morbidity as it expands to quality of life. It is within this framework that a study on childhood health in Jamaica is of vital importance. To (1) expand the health literature in Jamaica and by extension the Caribbean, (2) understand the status of child health outside of mortality, (3) aid public health practitioners with research, upon which they are able to further improve the quality of life of children by adding quality to their lived years, (4) investigate the age at with children in Jamaica become influenced by chronic disease, it typology, and (5) evaluate the subjective wellbeing of children as is done for the general populace and elderly. The current study extracted a sample of 8,373 and 2,104 children 0-14 years from two surveys. Descriptive statistics were used to analyze the socio-demographic characteristics of the sample. Data were analyzed using Chi-square and analysis of variance and independent sample t-test. The current study found that a shift in health condition was noticed in 2007 over 2002. The number of children who had diarrhea fell by 84.2% in 2007 over 2002, and a similar reduction was observed for those with asthma (42.1% in 2002 and 19.7% in 2007). Another critical finding was that 1.2% of children, in 2007, had diabetes mellitus. Public health now has an epidemiological profile of health conditions of children and the demographic shifts, which are occurring, and this can be used for effective management and planning of the new health reality of the Jamaican child.


Depression symptom outcomes and re-engagement among VA patients who discontinue care while symptomatic.
Evaluate outcomes of Veterans who discontinued treatment with at least moderate ongoing depressive symptoms. Veterans with elevated depression symptoms from 29 Department of Veterans Affairs facilities completed baseline surveys and follow-up assessments for one year. Analyses examined rates and predictors of treatment discontinuation, treatment re-engagement, and subsequent symptoms among patients who remained out of care. A total of 242 (17.8%; n = 1359) participants discontinued treatment while symptomatic, with Black participants, participants with less severe depression, and participants receiving only psychotherapy (versus combined psychotherapy and antidepressant medications) discontinuing at higher rates. Among all participants who discontinued treatment (n = 445), 45.8% re-engaged within the following six months with participants receiving combined treatment re-engaging at higher rates. Of participants who discontinued while symptomatic within the first 6 months of the study and did not return to care (n = 112), 68.8% remained symptomatic at 12 months. Lower baseline treatment expectancy and greater depression symptom severity were associated with remaining symptomatic while untreated. Black race, lower symptom severity, and treatment modality may help identify patients at higher risk for discontinuing care while symptomatic, whereas patients with lower treatment expectations may be at greater risk for remaining out of care despite continuing symptoms.


Mycobacterium smegmatis expressing a chimeric protein MPT64-proteolipid protein (PLP) 139-151 reorganizes the PLP-specific T cell repertoire favoring a CD8-mediated response and induces a relapsing experimental autoimmune encephalomyelitis.
We infected SJL mice with a recombinant Mycobacterium smegmatis expressing a chimeric protein containing the self-epitope of proteolipid protein 139-151 (p139) fused to MPT64, a secreted protein of Mycobacterium tuberculosis (rMS(p139)). Infected mice developed a relapsing experimental autoimmune encephalomyelitis (EAE), showing a prevailing demyelination of the CNS, and disease severity was significantly lower in comparison with the one that follows immunization with p139. rMS(p139) was not detected in lymph node or spleen in the course of clinical disease development or in the CNS during relapse. Infection with rMS(p139) modified the p139-specific T cell repertoire, recruiting the spontaneous p139-specific repertoire and activating CD4(+) T cells carrying the BV4 semiprivate rearrangement. T cells carrying the public BV10 rearrangement that are consistently found in the CNS during flares of disease were not activated by infection with rMS(p139) because lymph node APCs infected with rMS(p139) selectively fail to present the epitope for which BV10 cells are specific. Simultaneously, rMS(p139) expanded p139-specific CD8(+) cells more efficiently than immunization with peptide in adjuvant. SJL mice vaccinated against the CDR3 sequence of the BV10 public rearrangement reduced usage of the BV10 cells and displayed reduced symptoms during bouts of EAE. Thus, transient peripheral infection with a CNS-cross-reactive nonpathogenic Mycobacterium induces a relapsing EAE that continues long after clearance of the infectious agent. The composition of the self-reactive repertoire activated determines severity and histology of the resulting disease.


Effects of isosorbide mononitrate on the restoration of injured artery in mice in vivo.
The pharmacological basis of isosorbide mononitrate (ISMN), a widely used drug for cardiovascular diseases, is that it is metabolized to nitric oxide (NO). However, NO is a double-edged sword that results in either beneficial or detrimental effect. Vascular injury is the common consequence of many cardiovascular diseases, but it is not determined whether ISMN influences the restoration of injured artery in vivo. Carotid artery injury was induced by electric stimulation in mice. Vasoconstriction and endothelium-dependent and -independent relaxation were recorded by a multichannel acquisition and analysis system. ISMN (10 mg/kg, p.o.) treatment for 1 week and 1 month had no effect on reendothelialization, histology and function of carotid artery injured by electric stimulation. L-arginine (500 mg/kg, p.o.) and Nomega-nitro-L-arginine methyl ester (L-NAME) (50 mg/kg, p.o.) treatment for 1 week did not affect the reendothelialization process, but L-NAME treatment induced neointimal hyperplasia and inhibited endothelium-dependent relaxation in electrically injured artery. These results suggest that supplement of exogenous or endogenous NO has no effect on the restoration of injured artery, but inhibition of endogenous NO induces neointimal hyperplasia in injured artery. ISMN treatment does not affect the restoration of injured artery.


Advances in TERS (tip-enhanced Raman scattering) for biochemical applications.
TERS (tip-enhanced Raman scattering) provides exceptional spatial resolution without any need for labelling and has become a versatile tool for biochemical analysis. Two examples will be highlighted here. On the one hand, TERS measurements on a single mitochondrion are discussed, monitoring the oxidation state of the central iron ion of cytochrome c, leading towards a single protein characterization scheme in a natural environment. On the other hand, a novel approach of single molecule analysis is discussed, again based on TERS experiments on DNA and RNA, further highlighting the resolution capabilities of this method.


4-octyl itaconate ameliorates ventilator-induced lung injury.
Ventilator-induced lung injury (VILI) disturbs the disordered immune system and causes persistent inflammatory damage. 4-octyl itaconate (OI) is a synthetic cell-permeable itaconate derivative with antioxidant and anti-inflammatory effects. In this study, we assessed whether OI protects against VILI. OI was intraperitoneally injected for three days before mechanical ventilation (MV; 20 ml/kg at 70 breaths/min) for 2 h. Mouse lung vascular endothelial cells (MLVECs) were pretreated with OI (62.5, 125, and 250 μM) prior to cyclic stretch for 4 h. We found that OI attenuated VILI and inflammatory response. OI also increased superoxide dismutase, nuclear factor E2-related factor 2, and heme oxygenase-1 levels, and decreased reactive oxygen species and malondialdehyde levels. Furthermore, OI inhibited the expression of NLR family pyrin domain-containing 3 (NLRP3), caspase-1 p20, apoptosis-associated speck-like protein containing a CARD, and N-terminal fragment of gasdermin D. Therefore, OI attenuates VILI, potentially by suppressing oxidative stress and NLRP3 activation.


Transcranial direct current stimulation of the dorsolateral prefrontal cortex modulates repetition suppression to unfamiliar faces: an ERP study.
Repeated visual processing of an unfamiliar face suppresses neural activity in face-specific areas of the occipito-temporal cortex. This "repetition suppression" (RS) is a primitive mechanism involved in learning of unfamiliar faces, which can be detected through amplitude reduction of the N170 event-related potential (ERP). The dorsolateral prefrontal cortex (DLPFC) exerts top-down influence on early visual processing. However, its contribution to N170 RS and learning of unfamiliar faces remains unclear. Transcranial direct current stimulation (tDCS) transiently increases or decreases cortical excitability, as a function of polarity. We hypothesized that DLPFC excitability modulation by tDCS would cause polarity-dependent modulations of N170 RS during encoding of unfamiliar faces. tDCS-induced N170 RS enhancement would improve long-term recognition reaction time (RT) and/or accuracy rates, whereas N170 RS impairment would compromise recognition ability. Participants underwent three tDCS conditions in random order at ∼72 hour intervals: right anodal/left cathodal, right cathodal/left anodal and sham. Immediately following tDCS conditions, an EEG was recorded during encoding of unfamiliar faces for assessment of P100 and N170 visual ERPs. The P3a component was analyzed to detect prefrontal function modulation. Recognition tasks were administered ∼72 hours following encoding. Results indicate the right anodal/left cathodal condition facilitated N170 RS and induced larger P3a amplitudes, leading to faster recognition RT. Conversely, the right cathodal/left anodal condition caused N170 amplitude and RTs to increase, and a delay in P3a latency. These data demonstrate that DLPFC excitability modulation can influence early visual encoding of unfamiliar faces, highlighting the importance of DLPFC in basic learning mechanisms.


Reweaving the tapestry: a supertree of birds.
Our knowledge of the avian tree of life remains uncertain, particularly at deeper levels due to the rapid diversification early in their evolutionary history. They are the most abundant land vertebrate on the planet and have been of great historical interest to systematists. Birds are also economically and ecologically important and as a result are intensively studied, yet despite their importance and interest to humans around 13% of taxa currently on the endangered species list perhaps as a result of human activity. Despite all this no comprehensive phylogeny that includes both extinct and extant species currently exists. Here we present a species-level supertree, constructed using the Matrix Representation with Parsimony method, of Aves containing approximately two thirds of all species from nearly 1000 source phylogenies with a broad taxonomic coverage. The source data for the tree were collected and processed according to a strict protocol to ensure robust and accurate data handling. The resulting tree topology is largely consistent with molecular hypotheses of avian phylogeny. We identify areas that are in broad agreement with current views on avian systematics and also those that require further work. We also highlight the need for leaf-based support measures to enable the identification of rogue taxa in supertrees. This is a first attempt at a supertree of both extinct and extant birds, it is not intended to be utilised in an overhaul of avian systematics or as a basis for taxonomic re-classification but provides a strong basis on which to base further studies on macroevolution, conservation, biodiversity, comparative biology and character evolution, in particular the inclusion of fossils will allow the study of bird evolution and diversification throughout deep time.


Sensorimotor Convergence in Circuitry of the Motor Cortex.
Scientists and philosophers have long appreciated that active somatosensation requires the sensory and motor systems to exchange information about body the body's movements as well as touch in order to accurately interpret incoming somatosensory information and plan future movements. However, the circuitry underlying this sensory and motor integration is complicated and is difficult to study without tools to label specific cellular components in the various brain regions involved. Here, I review the general pathways that convey ascending sensory and descending motor information, using the rodent whisker system as a model to take advantage of the cell type specificity possible in this model. I then detail the circuits in motor cortex in which incoming information from somatosensory cortex and thalamus is integrated. I close with a brief description of changes in these circuits during motor learning.


Quantification of mouse mammary tumor virus structural proteins in hormone-induced mammary tumors of low mammary tumor mouse strains.
The expression of the mouse mammary tumor virus (MMTV) in hormone-induced mammary tumors was investigated by means of a radioimmunoassay for two major MMTV proteins, gp52 and p27. MMTV proteins were isolated on lectin affinity- and ion-exchange chromatography columns. The purified viral proteins were electrophoretically homogeneous and retained immunoreactivity after labelling with 125iodine. Standard competition assays showed that group-specific antigenic determinants were reacting. Mammary tumors were induced in three strains of mice with a low natural incidence of mammary tumors, C57BL, O20 and C3Hf, by a combined hormone treatment, consisting of hypophyseal isografts and administration of progesterone and estrone. Mammary tumors and mammary glands of hormone-treated animals were extracted and used for competition radioimmunoassays. In general, the tumorigenic hormone treatment resulted in enhanced amounts of MMTV proteins in the mammary glands, compared to the amounts found in lactating mammary glands of untreated animals. The levels of MMTV proteins in the mammary tumors were lower than in the mammary glands.


Antagonistic interplay of Swi1 and Tup1 on filamentous growth of Candida albicans.
Candida albicans is a polymorphic human opportunistic pathogen in which the Swi-Snf complex functions as an activator whereas Tup1 acts as a general repressor during the yeast-hyphae transition. In Saccharomyces cerevisiae, the interplay between the Swi-Snf complex and the Tup1-Ssn6 repressive complex regulates the balance between active and repressed chromatin structures of a number of genes. To study the interplay between Candida albicans Swi1 and Tup1 and their effects on morphogenesis, we analyzed phenotypes of swi1/swi1, tup1/tup1 and swi1/swi1 tup1/tup1 mutants under various growth conditions. The swi1/swi1 mutant failed to form true hyphae, whereas the tup1/tup1 mutant exhibited constitutive filamentous growth. Deletion of SWI1 in the tup1/tup1 mutant completely blocked hyphal growth under all the conditions examined. Under aerobic conditions, the swi1/swi1 tup1/tup1 mutant most resembled the swi1/swi1 mutant in phenotype, actin polarization and gene expression pattern. In invaded agar, the double mutant showed similar phenotypes as the swi1/swi1 mutant, while under embedded conditions, it grew as a pseudohypha-like form different from that of the wild-type strain, swi1/swi1 or tup1/tup1 mutants. These results suggest that Swi1 may play a dominant role by antagonizing the repressive effect of the Tup1 on hyphal development in C. albicans.


Epithelial cell binding of group A streptococci by lipoteichoic acid on fimbriae denuded of M protein.
Group A streptococci were treated with various enzymatic and chemical agents in an attempt to dissociate the type-specific M protein from intact surface "fimbriae." Mild peptic digestion at pH 5.8, which was previously shown to extract serologically active M antigen from intact streptococci had little visible effect on the fimbriae even though virtually all of the M protein was removed as demonstrated by (a) increased susceptibility to phagocytosis, (b) lack of opsonic effect of homologous M antibody on the treated streptococci, and (c) loss of HCl-extractable M protein. These fimbriated streptococci which lacked M protein adhered to human oral mucosal cells equally as well as untreated, fimbriated organisms which retained their M protein. Removal of both fimbriae and M protein by digesting organisms with HCL at pH 2.0 at 94 degrees C. or with trypsin abolished their ability to bind mucosal cells. Electron microscopy of streptococci bound to epithelial cells demonstrated fimbriae radiating from the surface of the organisms to the membrane of the epithelial cells. It is apparent, therefore, that the determinants of streptococcal fimbriae involved in resistance to phagocytosis can be dissociated from those involved in epithelial cell binding. These results are consistent with our previous studies which suggested that fatty acids ester linked with glycerol teichoic acid rather than M protein of streptococci binds the organisms to epithelial cells.


Dynamic characteristics of blood glucose time series during the course of critical illness: effects of intensive insulin therapy and relative association with mortality.
To assess the effect of intensive insulin therapy on blood glucose amplitude variation and pattern irregularity in critically ill patients. To assess the association of these blood glucose signal characteristics with hospital mortality, independent of blood glucose level. Retrospective analysis of the databases of two previously published randomized controlled trials. University hospital, 56-bed adult surgical intensive care unit and 17-bed medical intensive care unit. One thousand five-hundred forty-eight surgical intensive care unit patients, admitted between February 2000 and January 2001, and 1200 medical intensive care unit patients, admitted between March 2002 and May 2005. In the two randomized controlled trials, patients were randomized to receive either intensive insulin therapy (targeting normoglycemia, between 4.4 and 6.1 mmol/L) or conventional insulin therapy (infusing insulin when blood glucose levels were >12 mmol/L and stopping at 10 mmol/L). Intensive insulin therapy significantly lowered mean blood glucose (5.8 vs. 8.4 mmol/L), hyperglycemic index (0.8 vs. 3.2 mmol/L), and glycemic penalty index (26 vs. 53), but it increased the mean daily difference between minimum and maximum blood glucose (mean daily delta blood glucose; 4.0 vs. 3.3 mmol/L). There was no significant effect on the standard deviation of the blood glucose measurements or on jack-knifed approximate entropy. In multivariable logistic regression analysis, corrected for baseline risk factors, blood glucose levels outside the normoglycemic range, higher mean daily delta blood glucose, higher standard deviation blood glucose, and higher jack-knifed approximate entropy were independently associated with hospital mortality. The Leuven intensive insulin therapy strategy increased mean daily delta blood glucose while not affecting standard deviation blood glucose and jack-knifed approximate entropy. Increased blood glucose amplitude variation and pattern irregularity were associated with mortality, irrespective of blood glucose level. The reduced mortality observed with intensive insulin therapy in the Leuven trials cannot be attributed to an effect on blood glucose amplitude variation or entropy. Reducing amplitude variation and entropy of the blood glucose signal, irrespective of blood glucose concentration, may produce clinical benefits.


Alterations of the lung methylome in allergic airway hyper-responsiveness.
Asthma is a chronic airway disorder characterized by recurrent attacks of breathlessness and wheezing, affecting 300 million people around the world (available at: www.who.int). To date, genetic factors associated with asthma susceptibility have been unable to explain the full etiology of asthma. Recent studies have demonstrated that the epigenetic disruption of gene expression plays an equally important role in the development of asthma through interaction with our environment. We sensitized 6-week-old C57BL/6J mice with house-dust-mite (HDM) extracts intraperitoneally followed by 5 weeks of exposure to HDM challenges (three times a week) intratracheally. HDM-exposed mice showed an increase in airway hyper-responsiveness (AHR) and inflammation together with structural remodeling of the airways. We applied methylated DNA immunoprecipitation-next generation sequencing (MeDIP-seq) for profiling of DNA methylation changes in the lungs in response to HDM. We observed about 20 million reads by a single-run of massive parallel sequencing. We performed bioinformatics and pathway analysis on the raw sequencing data to identify differentially methylated candidate genes in HDM-exposed mice. Specifically, we have revealed that the transforming growth factor beta signaling pathway is epigenetically modulated by chronic exposure to HDM. Here, we demonstrated that a specific allergen may play a role in AHR through an epigenetic mechanism by disrupting the expression of genes in lungs that might be involved in airway inflammation and remodeling. Our findings provide new insights into the potential mechanisms by which environmental allergens induce allergic asthma and such insights may assist in the development of novel preventive and therapeutic options for this debilitative disease.


Dynamic metastable long-living droplets formed by sticker-spacer proteins.
Multivalent biopolymers phase separate into membrane-less organelles (MLOs) which exhibit liquid-like behavior. Here, we explore formation of prototypical MOs from multivalent proteins on various time and length scales and show that the kinetically arrested metastable multi-droplet state is a dynamic outcome of the interplay between two competing processes: a diffusion-limited encounter between proteins, and the exhaustion of available valencies within smaller clusters. Clusters with satisfied valencies cannot coalesce readily, resulting in metastable, long-living droplets. In the regime of dense clusters akin to phase-separation, we observe co-existing assemblies, in contrast to the single, large equilibrium-like cluster. A system-spanning network encompassing all multivalent proteins was only observed at high concentrations and large interaction valencies. In the regime favoring large clusters, we observe a slow-down in the dynamics of the condensed phase, potentially resulting in loss of function. Therefore, metastability could be a hallmark of dynamic functional droplets formed by sticker-spacer proteins.


Niche-specificity factors of a marine oil-degrading bacterium Alcanivorax borkumensis SK2.
Alcanivorax borkumensis strain SK2 is a cosmopolitan hydrocarbonoclastic marine bacterium, with a specialized metabolism adapted to the degradation of petroleum oil hydrocarbons. Transposon mutagenesis was used for functional genome analysis of Alcanivorax SK2 to reveal the genetic basis of other environmentally relevant phenotypes, such as biofilm formation, adaptation to UV exposure, and to growth at either low temperature or high salinity. Forty-eight relevant transposon mutants deficient in any one of these environmentally responsive functions were isolated, and the corresponding genes interrupted by the mini-Tn5 element were sequenced using inverse PCR. Several cross connections between different phenotypes (e.g. biofilm and UV stress; biofilm and UV and osmoadaptation) on signal transduction level have been revealed, pointing at complex and tightly controlled cellular interactions involving oxygen as a primary messenger and cyclic-di-GMP as a secondary messenger required for Alcanivorax responses to environmental stresses. These results provide insights into bacterial function in a complex marine environment.


Basal and stimulated sympathetic responses after epinephrine: no evidence of augmented responses.
Delayed facilitation of norepinephrine release through the action of epinephrine (NE) at presynaptic beta-adrenoceptors has been postulated to account for the delayed hemodynamic effects of epinephrine and to be a mechanism causally related to the development of hypertension. To determine whether a short-term increase in epinephrine concentrations resulted in subsequent facilitation of sympathetic responses, 9 healthy subjects (age, 21+/-0.9 years) were studied at rest and during physiological stress on 2 occasions when they received an infusion of either saline or epinephrine (20 ng/kg per minute) in random order. Heart rate, blood pressure, forearm blood flow, epinephrine concentrations, and NE spillover were measured at rest, during mental stress (Stroop test), and during a cold pressor test. Measurements were performed before, during the 1-hour infusion of epinephrine or placebo, and 1 hour after the infusion. A radioisotope dilution method was used to measure NE spillover. Hemodynamic measurements and NE spillover were increased during the infusion of epinephrine, but 1 hour after discontinuation of epinephrine there was no significant augmentation of hemodynamic or sympathetic responses. NE spillover 1 hour after saline or epinephrine infusion was similar (0.85+/-0.2 versus 0. 87+/-0.2 microg/min; P=0.92). In addition, there was no delayed facilitation of stress-induced hemodynamic or NE responses after epinephrine. These findings do not support the hypothesis that epinephrine results in delayed facilitation of NE release.


Pulmonary tumor thrombotic microangiopathy in patients with low-grade ovarian serous neoplasm: a clinicopathologic review of 2 cases of a previously unknown association.
Pulmonary tumor thrombotic microangiopathy (PTTM) is a rare complication occurring during tumor dissemination that can lead to severe, commonly unrecognized pulmonary hypertension, right-sided heart failure, and sudden death. Histologically, it is characterized by tumor microthrombi within small arteries and arterioles and associated fibrocellular and fibromuscular intimal proliferation. Gastric adenocarcinoma is the most common tumor type with this association. Of gynecologic malignancies, a single case of ovarian clear cell carcinoma has been linked to PTTM. We report 2 patients who underwent surgery with a preoperative diagnosis of pelvic mass. After surgery, the patients had unexplained progressive respiratory failure that led to their death. Autopsy revealed typical features of PTTM with tumor lymphangitic spread and microscopic tumor emboli within the lung arteries. In both cases, the primary tumor was an ovarian serous neoplasm of low malignant potential with widespread dissemination, 1 with microinvasion and progression to low-grade serous carcinoma. In this last case, mutational analysis for KRAS and BRAF genes was performed to confirm the association between the ovarian and the extraovarian tumor and rule out other primary tumors more commonly associated with this disease. PTTM is a distinct pathologic entity with very few cases reported in the literature, especially involving ovarian tumors. We report 2 cases of low-grade ovarian serous neoplasm, not previously reported with this association.


Mutations in genes lpxL1, bamA, and pmrB impair the susceptibility of cystic fibrosis strains of Pseudomonas aeruginosa to murepavadin.
Murepavadin is a peptidomimetic exhibiting specific inhibitory activity against Pseudomonas species. In the present study, its in vitro activity was assessed on 230 cystic fibrosis (CF) strains of Pseudomonas aeruginosa isolated from 12 French hospitals, in comparison with 12 other antipseudomonal antibiotics. Although murepavadin is still in preclinical stage of development, 9.1% (n = 21) of strains had a minimum inhibitory concentration (MIC) >4 mg/L, a level at least 128-fold higher than the modal MIC value of the whole collection (≤0.06 mg/L). Whole-genome sequencing of these 21 strains along with more susceptible isogenic counterparts coexisting in the same patients revealed diverse mutations in genes involved in the synthesis (lpxL1 and lpxL2) or transport of lipopolysaccharides (bamA, lptD, and msbA), or encoding histidine kinases of two-component systems (pmrB and cbrA). Allelic replacement experiments with wild-type reference strain PAO1 confirmed that alteration of genes lpxL1, bamA, and/or pmrB can decrease the murepavadin susceptibility from 8- to 32-fold. Furthermore, we found that specific amino acid substitutions in histidine kinase PmrB (G188D, Q105P, and D45E) reduce the susceptibility of P. aeruginosa to murepavadin, colistin, and tobramycin, three antibiotics used or intended to be used (murepavadin) in aerosols to treat colonized CF patients. Whether colistin or tobramycin may select mutants resistant to murepavadin or the opposite needs to be addressed by clinical studies.


Monitoring activation of the antiviral pattern recognition receptors RIG-I and PKR by limited protease digestion and native PAGE.
Host defenses to virus infection are dependent on a rapid detection by pattern recognition receptors (PRRs) of the innate immune system. In the cytoplasm, the PRRs RIG-I and PKR bind to specific viral RNA ligands. This first mediates conformational switching and oligomerization, and then enables activation of an antiviral interferon response. While methods to measure antiviral host gene expression are well established, methods to directly monitor the activation states of RIG-I and PKR are only partially and less well established. Here, we describe two methods to monitor RIG-I and PKR stimulation upon infection with an established interferon inducer, the Rift Valley fever virus mutant clone 13 (Cl 13). Limited trypsin digestion allows to analyze alterations in protease sensitivity, indicating conformational changes of the PRRs. Trypsin digestion of lysates from mock infected cells results in a rapid degradation of RIG-I and PKR, whereas Cl 13 infection leads to the emergence of a protease-resistant RIG-I fragment. Also PKR shows a virus-induced partial resistance to trypsin digestion, which coincides with its hallmark phosphorylation at Thr 446. The formation of RIG-I and PKR oligomers was validated by native polyacrylamide gel electrophoresis (PAGE). Upon infection, there is a strong accumulation of RIG-I and PKR oligomeric complexes, whereas these proteins remained as monomers in mock infected samples. Limited protease digestion and native PAGE, both coupled to western blot analysis, allow a sensitive and direct measurement of two diverse steps of RIG-I and PKR activation. These techniques are relatively easy and quick to perform and do not require expensive equipment.


Discriminative Sparse Neighbor Approximation for Imbalanced Learning.
Data imbalance is common in many vision tasks where one or more classes are rare. Without addressing this issue, conventional methods tend to be biased toward the majority class with poor predictive accuracy for the minority class. These methods further deteriorate on small, imbalanced data that have a large degree of class overlap. In this paper, we propose a novel discriminative sparse neighbor approximation (DSNA) method to ameliorate the effect of class-imbalance during prediction. Specifically, given a test sample, we first traverse it through a cost-sensitive decision forest to collect a good subset of training examples in its local neighborhood. Then, we generate from this subset several class-discriminating but overlapping clusters and model each as an affine subspace. From these subspaces, the proposed DSNA iteratively seeks an optimal approximation of the test sample and outputs an unbiased prediction. We show that our method not only effectively mitigates the imbalance issue, but also allows the prediction to extrapolate to unseen data. The latter capability is crucial for achieving accurate prediction on small data set with limited samples. The proposed imbalanced learning method can be applied to both classification and regression tasks at a wide range of imbalance levels. It significantly outperforms the state-of-the-art methods that do not possess an imbalance handling mechanism, and is found to perform comparably or even better than recent deep learning methods by using hand-crafted features only.


The antiretroviral drug pipeline: prospects and implications for future treatment research.
A number of investigational antiretroviral drugs in clinical development could alter the future treatment landscape for resource-limited settings and contribute to optimized therapy for HIV infection. Several nucleoside reverse transcriptase inhibitors (NRTIs) are in development, including festinavir (BMS-986001), a thymidine analogue similar to stavudine but with reduced potential for toxicity, CMX-157, a hexadecyloxypropyl conjugate of tenofovir and tenofovir alafenamide (GS-7340), a prodrug of tenofovir achieving much higher intracellular triphosphate concentrations with a lower dose than tenofovir disoproxil fumarate. MK-1439 is a well tolerated once-daily non-NRTI (NNRTI) with activity against most NNRTI-associated resistance mutations. Albuvirtide is a long-acting parenteral fusion inhibitor related to enfuvirtide, and BMS-663068 is an oral HIV attachment/entry inhibitor. Ibalizumab (formerly TNX-355) is an mAb that binds to CD4 and lowers HIV plasma viral RNA in infected patients. Cenicriviroc is a CCR5-antagonist that also has activity against the inflammatory chemokine CCR2. The integrase strand transfer inhibitor (InSTI) dolutegravir was recently approved in the U.S. and is an attractive component of future regimens because of efficacy, tolerability, apparent safety and once-daily dosing; it also maintains some activity against raltegravir and elvitegravir-resistant mutants. The dolutegravir analogue GSK-1265744 appears to be equipotent and is being developed as a long-lasting injectable parenteral agent. The selective cytochrome P450 3A4 inhibitor cobicistat is a better tolerated alternative to ritonavir for pharmacokinetic 'boosting', but may also result in clinically undesirable drug interactions. There are several investigational antiretroviral drugs with significant promise for inclusion in future primary and secondary combination regimens.


[Effect of fluoroquinolones made in China in the treatment of acute bacillary dysentery. A report of 212 cases].
212 cases of acute bacillary dysentery were treated with enoxacin and norfloxacin, another 15 cases were treated by chujunsheng. The cure rate in enoxacin group was 99.1%, in norfloxacin group 98.04% and in chujunsheng group 63.64%. The isolated pathogens were tested for susceptibility with disk diffusion and MIC determination. The results showed that both enoxacin and norfloxacin are better than gentamycin, ampicillin, chloramphenicol and trimethoprim-sulfamethoxazole. Side effects of enoxacin group and norfloxacin were noted in are 2.7% and 3.0% of the patients respectively. This study suggests that enoxacin and norfloxacin are highly effective, quite convenient for use and have few adverse reactions. They are considered to be drugs of choice for the treatment of acute bacillary dysentery.


@neuLink: a service-oriented application for biomedical knowledge discovery.
We introduce the architecture of @neuLink, a service-oriented environment for biomedical knowledge discovery which has been developed in the course of EU Integrated Project @neurIST. The application integrates data from databases with information extracted from unstructured text sources. Moreover, @neuLink supports the analysis of primary biomolecular data associated with individual patients and thus enables the interpretation of molecular data inside a clinical research environment. Based on an assembly of data services, @neuLink interacts with the complex @neurIST grid infrastructure through a dedicated data access and data mediation service. Data types integrated by @neuLink are covering the entire span of biomolecular entities: from gene names in text to entries in EntrezGene; from mentions of drugs to Drugbank, from information on allelic variants in scientific literature to entries in dbSNP. The architecture of @neuLink allows easy integration of other webservice-based applications and thus the spectrum of analysis capabilities of @neuLink can be extended following the requirements of the users of the @neurIST system.


Ghrelin: new insight to mechanisms and treatment of postoperative gastric ileus.
Postoperative gastric ileus develops after abdominal surgery and if prolonged leads to longer hospitalization times. Besides discomfort for the patient such as abdominal bloating and pain, this condition is associated with a great increase in healthcare costs. In order to develop new and effective treatment strategies to alleviate the ileus, a good understanding of the pathophysiogical underlying mechanisms is necessary. Postoperative gastric ileus consists of two phases, a first neural/humoral phase and a second inflammatory phase. The present review will focus on the role of the orexigenic and prokinetic hormone ghrelin in the development of postoperative ileus. Although ghrelin and ghrelin mimetics have been used during the recent years in pre-clinical and a first clinical trial to alleviate symptoms of postoperative ileus, the regulation of this hormone under conditions of abdominal surgery has just recently begun to be explored. Growing evidence indicates that postoperative restoration of ghrelin signaling leads to an orexigenic effect whereas the gastroprokinetic actions seem to require supraphysiological doses. Since the expression of ghrelin-O-acyltransferase (GOAT), the only ghrelin acylating enzyme, is reduced after abdominal surgery, the stimulation of GOAT signaling may also be a new promising approach to restore levels of acyl ghrelin under these conditions and a potential useful addition to ghrelin mimetics.


On wakefulness fluctuations as a source of BOLD functional connectivity dynamics.
Human brain dynamics and functional connectivity fluctuate over a range of temporal scales in coordination with internal states and environmental demands. However, the neurobiological significance and consequences of functional connectivity dynamics during rest have not yet been established. We show that the coarse-grained clustering of whole-brain dynamic connectivity measured with magnetic resonance imaging reveals discrete patterns (dynamic connectivity states) associated with wakefulness and sleep. We validate this using EEG in healthy subjects and patients with narcolepsy and by matching our results with previous findings in a large collaborative database. We also show that drowsiness may account for previous reports of metastable connectivity states associated with different levels of functional integration. This implies that future studies of transient functional connectivity must independently monitor wakefulness. We conclude that a possible neurobiological significance of dynamic connectivity states, computed at a sufficiently coarse temporal scale, is that of fluctuations in wakefulness.


[Postgraduate medical education in Iceland; medical students' and residents' attitude].
Various parties have expressed interest in establishing formal postgraduate medical education programs. The interest of residents and medical students to such programs in Iceland has not been evaluated before. A questionnaire was sent to 146 interns and residents and 84 senior medical students. The following variables were analyzed: Gender, attitude towards postgraduate training in Iceland, interest to participate in such training, preferred specialty and which factors might influence their decision. 100 subjects completed the questionnaire (response rate 45%), 61 interns and residents and 39 medical students. Of those completing the questionnaire, most interns and residents (97%) and medical students (87%) agreed or agreed strongly with the concept of postgraduate training in Iceland. The majority of responders wanted to undergo part of their postgraduate training in Iceland if this option would be available. Those who preferred postgraduate training in Iceland did so for the following reasons: The importance of "hands-on" training, availability of consultants, favorable social setting and organized teaching. Those who wanted to do all their postgraduate training abroad rated case variability, "hands-on" training, organized teaching and research opportunities as the main factors influencing their decision. Interns, residents and medical students are in favor of undergoing part of their postgraduate training in Iceland. Those who chose to train abroad rated case variability and research opportunities higher than those in favor of training in Iceland.


Simple confirmation methods for rare but impaired variants of human flavin-containing monooxygenase 3 (FMO3) found in an updated genome resource databank.
Forty-seven new nonsense or missense human flavin-containing monooxygenase 3 (FMO3) variants were recently identified in an updated Japanese population reference panel. Of these, 20 rare single-nucleotide substitutions resulted in moderately or severely impaired FMO3 activity. To easily identify these 20 FMO3 variants (2 stop codon mutations, 2 frameshifts, and 16 amino-acid substitutions) in the clinical setting, simple confirmation methods for impaired FMO3 variants are proposed using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) or allele-specific PCR methods. Using PCR-RFLP, FMO3 variants p.Arg51Gly, p.Met66Lys, p.Asn80Lys, p.Val151Glu, p.Val187fsTer25, p.Gly193Arg, p.Val283Ala, p.Asp286His, p.Val382Ala, and p.Phe451Leu were digested by the designated restriction enzymes and confirmed using reference cDNAs. In contrast, the FMO3 variants p.Gly39Val, p.Arg238Ter, p.Arg387Cys, p.Arg387His, p.Leu457Trp, and p.Met497Arg were not digested, whereas the wild type was digested. FMO3 variants p.Gly11Asp, p.Lys416fsTer72, p.Gln427Ter, and p.Thr453Pro were confirmed using allele-specific PCR systems. The previously identified FMO3 p.Arg500Ter variant has a relatively high frequency and was differentiated from p.Arg500Gln in two steps, i.e., enzyme restriction followed by allele-specific PCR, similar to the method for p.Arg387Cys and p.Arg387His. These systems should facilitate easy detection in the clinical setting of FMO3 variants in Japanese subjects susceptible to low drug clearance possibly caused by impaired FMO3 function.


Greater mortality risk among patients with delayed follow-up after implantable cardioverter defibrillator procedures.
Substantial numbers of deaths occur among implantable cardioverter defibrillator (ICD) recipients. Timely follow-up after ICD implantation might improve patient outcomes. Our objective was to examine follow-up care and outcomes among patients undergoing ICD implantation. We compared patient characteristics and outcomes of those with late (> 12 weeks) vs early defibrillator clinic follow-up after ICD implantation in the Ontario ICD Database from 2007 to 2011. We examined the effect of ICD clinic follow-up visits after implant compared with primary care physician tracer on mortality outcomes using time-varying covariate analyses. Among 8096 ICD recipients (age 64.6 ± 12.6 years), 1145 (14%) received delayed follow-up. Patients with early ICD clinic follow-up experienced reduced risk of all-cause mortality, with a hazard ratio [HR] of 0.69 (95% confidence interval [CI], 0.50-0.95; P = 0.023) adjusted for clinical factors, primary care visits after discharge, and distance to follow-up centre. Early ICD clinic follow-up was associated with a reduction in out-of-hospital death with adjusted HR of 0.52 (95% CI, 0.36-0.76; P = 0.001) but not with a significant change in cardiovascular hospitalizations. In a tracer analysis, early primary care physician follow-up was associated with a nonsignificant trend toward increased mortality with an adjusted HR of 1.48 (95% CI, 0.97-2.25; P = 0.072). Reduced estimated glomerular filtration rate, secondary prevention or replacement devices, myocardial ischemia, smoking, and greater geographic distance to the implanting centre increased the odds of delayed follow-up. After device implantation, early defibrillator clinic follow-up was associated with reduced risk of all-cause and out-of-hospital death compared with those experiencing delayed follow-up.


Molecular pathways: PI3K pathway phosphatases as biomarkers for cancer prognosis and therapy.
Cancer research has seen tremendous changes over the past decade. Fast progress in sequencing technology has afforded us with landmark genetic alterations, which had immediate impact on clinical science and practice by pointing to new kinase targets, such as phosphoinositide 3-kinase (PI3K), the EGF receptor, or BRAF. The PI3K pathway for growth control has emerged as a prime example for both oncogene activation and tumor suppressor loss in cancer. Here, we discuss how therapy using PI3K pathway inhibitors could benefit from information on specific phosphatases, which naturally antagonize the kinase targets. This PI3K pathway is found mutated in most cancer types, including prostate, breast, colon, and brain tumors. The tumor-suppressing phosphatases operate at two levels. Lipid-level phosphatases, such as PTEN and INPP4B, revert PI3K activity to keep the lipid second messengers inactive. At the protein level, PHLPP1/2 protein phosphatases inactivate AKT kinase, thus antagonizing mTOR complex 2 activity. However, in contrast with their kinase counterparts the phosphatases are unlikely drug targets. They would need to be stimulated by therapy and are commonly deleted and mutated in cancer. Yet, because they occupy critical nodes in preventing cancer initiation and progression, the information on their status has tremendous potential in outcome prediction, and in matching the available kinase inhibitor repertoire with the right patients. Clin Cancer Res; 20(12); 3057-63. ©2014 AACR.


Long-term efficacy of dietary weight loss in sleep apnoea/hypopnoea syndrome.
Weight loss is associated with clinical improvement in sleep apnoea/ hypopnoea syndrome (SAHS). The aim of this study was to ascertain whether the therapeutic efficacy of dietary weight loss is maintained in the long-term. From a total of 216 overweight SAHS patients treated by only a weight reduction programme, 24 cured by this method (apnoea/hypopnoea index (AHI) at diagnosis 443+/-27.8, after weight loss 3+/-3.1) were re-evaluated after a mean (+/-SD) period of 94.3+/-27.4 months post-cure. No correlation was found between changes in AHI and body mass index experienced by each patient in the two phases of the study (diagnosis to cure and cure to long-term follow-up), r=0.29, p=0.156, demonstrating a marked intra-individual variability. Six of the 13 patients who maintained their weight presented recurrence of SAHS (AHI 40.5+/-24.1) as did eight of the 11 who had regained weight (AHI 44.3+/-23.). Weight maintenance was more frequent among patients who had continued to attend periodic appointments, 10/11 versus 3/13 (p<0.001). In conclusion, weight-loss efficacy is maintained in the long-term in some sleep apnoea/hypopnoea syndrome patients. This study indicates the need for periodic follow-up of these patients as a reinforcement for weight maintenance and for early detection of the reappearance of sleep apnoea/hypopnoea syndrome.


[An experience with stenting the esophagus in patients with inoperable cancer].
An experience with treatment of 21 patients with inoperable cancer of the esophagus allowed determination of indications and contraindications, development of the method and technique of endoscopic implantation of a domestic meshed self-expanding stent. The complications resulting from the process of mounting and the stent itself are analyzed. The methods of eliminating the complications are shown. The immediate and nearest results of endoprosthesing are assessed. Recommendations are given concerned with the cases of ingrowth of tumor into the stent.


Tuning the mechanical properties of nanoporous hydrogel particles via polymer cross-linking.
Soft hydrogel particles with tunable mechanical properties are promising for next-generation therapeutic applications. This is due to the increasingly proven role that physicochemical properties play in particulate-based delivery vectors, both in vitro and in vivo. The ability to understand and quantify the mechanical properties of such systems is therefore essential to optimize function and performance. We report control over the mechanical properties of poly(methacrylic acid) (PMA) hydrogel particles based on a mesoporous silica templating method. The mechanical properties of the obtained particles can be finely tuned through variation of the cross-linker concentration, which is hereby quantified using a cross-linking polymer with a fluorescent tag. We demonstrate that the mechanical properties of the particles can be elucidated using an atomic force microscopy (AFM) force spectroscopy method, which additionally allows for the study of hydrogel material properties at the nanoscale through high-resolution force mapping. Young's modulus and stiffness of the particles were tuned between 0.04 and 2.53 MPa and between 1.6 and 28.4 mN m(-1), respectively, through control over the cross-linker concentration. The relationship between the concentration of the cross-linker added and the amount of adsorbed polymer was observed to follow a Langmuir isotherm, and this relationship was found to correlate linearly with the particle mechanical properties.


Time trend by region of suicides and suicidal thoughts among Greenland Inuit.
Suicides remain a major public health problem in Greenland. Their increase coincides with the modernization since 1950. Serious suicidal thoughts are reported by a significant proportion of participants in countrywide surveys. To analyze the time trend by region of suicides and suicidal thoughts among the Inuit in Greenland. Data included the Greenland registry of causes of death for 1970-2011 and 2 cross-sectional health surveys carried out in 1993-1994 and 2005-2010 with 1,580 and 3,102 Inuit participants, respectively. Suicide rates were higher among men than women while the prevalence of suicidal thoughts was higher among women. Suicide rates for men and women together increased from 1960 to 1980 and have remained around 100 per 100,000 person-years since then. The regional pattern of time trend for suicide rates varied with an early peak in the capital, a continued increase to very high rates in remote East and North Greenland and a slow increase in villages relative to towns on the West Coast. Suicidal thoughts followed the regional pattern for completed suicides. Especially for women there was a noticeable increasing trend in the villages. The relative risk for suicide was highest among those who reported suicidal thoughts, but most suicides happened outside this high-risk group. Suicide rates and the prevalence of suicidal thoughts remain high in Greenland but different regional trends point towards an increased marginalization between towns on the central West Coast, villages and East and North Greenland. Different temporal patterns call for different regional strategies of prevention.


Kinetics of early and late spleen colony development.
The kinetics of spleen colony development were studied with special emphasis upon the self-replicative capacity of CFU-S, which are responsible for the early- or late-appearing spleen colonies, and upon CFU-S migration in the days following transplantation. It was found that, while the spleen colonies had a much lower daughter CFU-S content at eight days than at 11 or 12 days, many of the 8-day colonies remained in the spleen to become 12-day colonies which had many daughter CFU-S. In addition, it was found that large numbers of CFU-S were present in the splenic space between the colonies. Evidence was developed that these "intercolonial space" CFU-S were the founders of many of the late-appearing spleen colonies and that many or all of them were transported from the bone marrow to the spleen via the blood at varying times after the transplantation. Approximately one-half of the day-12 colony founders were found to arrive in the spleen after the third posttransplantation day.


An assay for selection of sera with circulating Toxoplasma gondii antigens.
We have developed an ELISA that employs monoclonal anti-Toxoplasma SAG1 (p30) as the capture antibody to detect T. gondii circulating antigens in patients' serum samples. Using serum spiked with Toxoplasma soluble and with SAG1 recombinant proteins, the detection limits were 31.25 ng/mL and 62.50 ng/mL, respectively. We obtained positive results in 28% (21/75) and 11% (23/206) of probable active and chronic toxoplasmosis serum samples, respectively. Western blot analysis on pooled antigen-positive serum samples showed antigenic bands of molecular weights 25 and 75 kDa from sera of probable active infection and five antigenic bands ranging in size from 26 to 33 kDa from chronic infection sera. This assay would be useful as an initial serum selection step in developing a Toxoplasma antigen detection test and for characterization studies.


Properties of the 40 kDa antigen of Mycobacterium tuberculosis, a functional L-alanine dehydrogenase.
The 40 kDa antigen of Mycobacterium tuberculosis is the first antigen reported to be present in the pathogenic M. tuberculosis, but not in the vaccine strain Mycobacterium bovis BCG. It is a functional L-alanine dehydrogenase (EC 22.214.171.124) and hence one of the few antigens possessing an enzymic activity. This makes the 40 kDa antigen attractive for potential diagnostic and therapeutic interventions. Recently, we developed a strategy to purify quantities of the recombinant protein in active form, and here we describe the biochemical properties of this enzyme. In the oxidative-deamination reaction, the enzyme showed K(m) values of 13. 8 mM and 0.31 mM for L-alanine and NAD(+), respectively, in a random-ordered mechanism. K(m, app) values in the reductive-amination reaction are 35.4 mM, 1.45 mM and 98.2 microM for ammonium, pyruvate and NADH, respectively. The enzyme is highly specific for all of its substrates in both directions. The pH profile indicates that oxidative deamination virtually may not occur at physiological pH. Hence L-alanine most likely is the product of the reaction catalysed in vivo. The enzyme is heat-stable, losing practically no activity at 60 degrees C for several hours.


Diagnostic Utility of Urinary LTE4 in Asthma, Allergic Rhinitis, Chronic Rhinosinusitis, Nasal Polyps, and Aspirin Sensitivity.
Urinary leukotriene E4 (LTE4) is a well-validated marker of the cysteinyl leukotriene pathway, and LTE4 elevation has been described in conditions such as asthma, aspirin sensitivity, and chronic rhinosinusitis (CRS). There have been a number of reports investigating the role of spot urine LTE4 to predict aspirin sensitivity; however, variability in urinary LTE4 may affect the accuracy of this approach. Here, we explored the utility of 24-hour urinary LTE4 in 5 clinical diagnoses of allergic rhinitis, asthma, chronic rhinosinusitis with nasal polyps (CRSwNP), CRS without nasal polyps, and aspirin sensitivity. This was a retrospective review of patients who had 24-hour quantification of urinary LTE4 by a clinically validated liquid chromatography tandem mass spectrometry method and their assigned diagnoses after assessment and clinical care. Twenty-four-hour urinary LTE4 elevations were seen in those with asthma and those with CRSwNP but influenced by underlying aspirin sensitivity. Elevation in LTE4 was significant in those with CRSwNP after adjusting for aspirin sensitivity. Allergic rhinitis was not associated with elevated LTE4 excretion. Receiver operator characteristic analysis of 24-hour urinary LTE4 showed that a cutoff value of 166 pg/mg Cr suggested the presence of history of aspirin sensitivity with 89% specificity, whereas a cutoff value of 241 pg/mg Cr discriminated "challenge-confirmed" aspirin-sensitive subjects with 92% specificity. Elevated 24-hour excretion of urinary LTE4 is a reliable and simple test to identify aspirin sensitivity in patients with respiratory diagnoses.


Measuring The Impact Of Air Pollution On Health Care Costs.
Air pollution contributes to the development of numerous adverse human health outcomes. The Environmental Protection Agency's Environmental Benefits Mapping and Analysis Program-Community Edition (BenMAP-CE) tool is widely used in estimating the health care costs of air pollution and in the development of federal and state regulations and policy. Its default features consider only the costs of hospital and emergency department admissions. A more complete accounting of the chain of costs would include ambulatory and other care. In this article we use employer health insurance claims data to infer additional costs that accompany hospitalizations but are not included in BenMAP-CE. Including additional categories increases BenMAP-CE health care cost estimates by approximately 40 percent for respiratory and cardiovascular patients. That is, for each dollar of health care costs captured by BenMAP-CE, a more complete accounting would include an additional 40 cents. These results suggest that because such air pollution costs are underestimated, the health care benefits associated with reducing air pollution may be much larger than previously estimated.


Comparison of four hemolysin-producing organisms (Escherichia coli, Serratia marcescens, Aeromonas hydrophila, and Listeria monocytogenes) for release of inflammatory mediators from various cells.
We investigated the role of various hemolysin-producing strains (Escherichia coli, Serratia marcescens, Aeromonas hydrophila, and Listeria monocytogenes) in induction of inflammatory mediators, e.g., histamine release from rat mast cells as well as the chemiluminescence response and the release of lipoxygenase transformation products from human polymorphonuclear neutrophils. Our data show that the hemolysin-positive bacteria as well as the hemolysin-positive culture supernatants were active in inducing the chemiluminescence response, leukotriene (LTB4 and LTC4) release from human granulocytes, and histamine release from rat mast cells. The degree of leukotriene release was dependent on the hemolysin type and on the expression of hemolysin activity. The E. coli alpha-hemolysin and the aerolysin-producing A. hydrophila were the most potent stimuli whether washed bacteria or bacterial supernatant was used. Bacteria expressing the S. marcescens hemolysin and the listeriolysin were only poor inducers of leukotriene generation. In contrast to leukotriene generation, all hemolysin-positive strains induced nearly the same histamine release in a dose-dependent manner. Our data suggest a potent role for various hemolysins as virulence factors in inducing the release of inflammatory mediators.


Suppression of eating by fenfluramine in patients with bulimia nervosa.
Fifteen patients with bulimia nervosa received fenfluramine (60 mg po) or placebo under double-blind, randomly ordered conditions. Two hours later food was presented. Significantly less food was eaten after fenfluramine and the quantity eaten was inversely correlated with serum fenfluramine levels. Significantly fewer patients reported bulimic symptoms during the test after fenfluramine, but no significant effect was demonstrated after leaving the ward. Fenfluramine caused drowsiness but did not reduce hunger ratings. Similarly, eating failed to reduce hunger ratings normally in the patients. These findings suggest that in patients with bulimia nervosa, hunger is reported abnormally and eating is suppressed by fenfluramine. Bulimic symptoms were probably reduced by fenfluramine, which may prove to be a useful treatment for bulimia nervosa.


Selective blockage of delta opioid receptors prevents the development of morphine tolerance and dependence in mice.
Recently, we demonstrated that delta opioid binding sites are involved in the development of morphine tolerance and dependence. In our present work, we studied the effect of the potent and selective delta antagonist, naltrindole (NTI), and its nonequilibrium analog, naltrindole 5'-isothiocyanate (5'-NTII), on the development of morphine tolerance and dependence in mice. In the acute model, mice injected with 100 mg/kg of morphine sulfate s.c. displayed acute tolerance 4 hr later as evidenced by a greater than 3-fold increase of the ED50 of morphine sulfate when compared to that of control mice. The acute tolerance was accompanied by the development of acute physical dependence as seen by the dramatic decrease in the amount of naloxone required to precipitate withdrawal jumping. Likewise, in the chronic model s.c. implantation of morphine pellets (75 mg free base) for 3 days produced tolerance and physical dependence. The ED50 of morphine sulfate in this case was increased by about 19-fold and the amount of naloxone needed to precipitate withdrawal jumping was 40 times lower than that required for acutely dependent mice. The development of acute tolerance and dependence was suppressed markedly in mice pretreated with NTI before induction of tolerance and dependence with 100 mg/kg of morphine sulfate. Multiple administration of either NTI or 5'-NTII before and during implantation with morphine base pellets also inhibited substantially the development of morphine tolerance and dependence.(ABSTRACT TRUNCATED AT 250 WORDS)


Purification and characterization of grass carp mitochondrial aldehyde dehydrogenase.
The molecular biology and enzymology of aldehyde dehydrogenase (ALDH) have been extensively investigated. However, most of the studies have been confined to the mammalian forms, while the sub-mammalian vertebrate ALDHs are relatively unexplored. In the present investigation, an ALDH was purified from the hepatopancreas of grass carp (Ctenopharygodon idellus) by affinity chromatographies on alpha-cyanocinnamate-Sepharose and Affi-gel Blue agarose. The 800-fold purified enzyme had a specific activity of 4.46 U/mg toward the oxidation of acetaldehyde at pH 9.5. It had a subunit molecular weight of 55000. Isoelectric focusing showed a single band with a pI of 5.3. N-terminal amino acid sequencing of 30 residues revealed a positional identity of approximately 70% with mammalian mitochondrial ALDH2. The kinetic properties of grass carp ALDH resembled those of mammalian ALDH2. The optimal pH for the oxidation of acetaldehyde was 9.5. The K(m) values for acetaldehyde were 0.36 and 0.31 microM at pH 7.5 and 9.5, respectively. Grass carp ALDH also possessed esterase activity which could be activated in the presence of NAD(+).


Pyoderma Gangrenosum: A Report of 27 Patients.
The aim of this study was to document the profile of pyoderma gangrenosum (PG) patients who were treated in our clinic and to compare our patients' clinical and demographic characteristics with those described in the literature to help our understanding of the total burden of PG. A total of 27 (17 women, 10 men) patients with a mean age 48.6 years at diagnosis were included in the study. Seven (25.9%) of them had 3 or more ulcers. The lower extremity was the most common site of occurrence (92.5%). During the study period, 6 patients were admitted twice, and 7 were admitted 3 or more times (range 1 to >10), and the median length of stay was 27 days. A concomitant disease was present in PG patients especially in those between 20 and 40 years of age. Systemic therapy was used in 21 (91.3%) patients, 17 patients were treated with systemic corticosteroids, either alone in 7 patients or combination with other agents in 10 patients. Three of our patients died because of disease or treatment-related complications during the study period. Despite the high (70.3%) percentage of the PG patients with improving disease at the time of discharge, 1 year after hospital discharge, one third of our 27 patients still had PG requiring readmission. In conclusion, none of the current therapies provide satisfying results in all of the patients, and our data indicate a long-standing unmet need for effective therapy for the unexpected course of PG.


[Dengue: a re-emerging disease. A clinical and epidemiological study in 57 Spanish travelers].
Dengue infection is nowadays considered a re-emergent disease. It has a worldwide tropical and subtropical distribution. The dengue virus in a member of the flavivirus family composed by 4 different serotypes. The virus is transmitted by mosquitos of the Aedes genus. With the increment of travels to the endemic areas, dengue is now observed frequently in our country. We analyzed 57 patients, 30 with imported dengue (ID) and 27 with dengue fever suffered during the trip (DDT). This series is compared with other published ones and a review of the subject is presented. Patients with ID followed a protocol as a febril syndrome returning from the tropics. Dengue was diagnosed through a compatible clinico-epidemiological history, the absence of other ferbil illness and positivity of specific serology. All patients had travelled to endemic areas (Central America 28 cases, Indian subcontinent 15, South-East Asia 10, South America 2, West Africa one, and Pacific one). The following were the most important clinical characteristics: fever and asthenia (100%), headache (98%), mialgia (84%), arthralgia (72%), morbilliform rash (61%) and retroocular pain (65%). For ID cases, the most helpful analitical results were: leucopenia (70%), reactive lymphocytes in peripheral blood smear (70%), thrombocytopenia (70%), and increased hepatic enzymes ALAT (53%), ASAT (63%) and LDH (100% in the 7 patients tested for this enzyme). Dengue must be included in differential diagnosis of fever in patients coming back to travels to tropical areas.


Traumatic dissection of the vertebral artery: a case report.
This report concerns a patient suffering from a severe neck pain on the third day after a traffic accident. This condition was followed by a lateral medullary infarction on the next day. One week later, he was transferred to this hospital and dissection of the vertebral artery was disclosed. It should be emphasized that in case of acute cervical spine injury or severe neck pain after a major trauma, vertebral artery dissection should be considered, as its early diagnosis may have crucial therapeutic implications. In addition, the advantages of the combination of magnetic resonance imaging and magnetic resonance angiography as diagnostic tools-of-choice are emphasized.


The development of fungal networks in complex environments.
Fungi are of fundamental importance in terrestrial ecosystems playing important roles in decomposition, nutrient cycling, plant symbiosis and pathogenesis, and have significant potential in several areas of environmental biotechnology such as biocontrol and bioremediation. In all of these contexts, the fungi are growing in environments exhibiting spatio-temporal nutritional and structural heterogeneities. In this work, a discrete mathematical model is derived that allows detailed understanding of how events at the hyphal level are influenced by the nature of various environmental heterogeneities. Mycelial growth and function is simulated in a range of environments including homogeneous conditions, nutritionally-heterogeneous conditions and structurally-heterogeneous environments, the latter emulating porous media such as soils. Our results provide further understanding of the crucial processes involved in fungal growth, nutrient translocation and concomitant functional consequences, e.g. acidification, and have implications for the biotechnological application of fungi.


Factor structure of the four motor-free scales of the Developmental Test of Visual Perception, 2nd edition (DTVP-2).
Visual-perceptual skills of children are often assessed using the Developmental Test of visual Perceptual Skills-2, 2nd Edition (DTVP-2). The factor structure of the four motor-free DTVP-2 subscales was examined. The scores from a sample of 356 healthy children (171 boys and 185 girls) ages 5 to 11 years were used to complete a principal components factor analysis of the 4 subscales. The position-in-space subscale had items load on six factors. The figure ground subscale had items load on five factors. The visual closure scale and form constancy subscales both had four factors. When the four DTVP-2 subscales were combined to form one overall motor-free visual perceptual scale, the items loaded on 21 factors. The DTVP-2 and its four motor-free subscales exhibited multidimensionality instead of the expected unidimensional visual-perceptual constructs.


[Therapy and prognosis of subarachnoid hemorrhage due to rupture of a cerebral aneurysm].
The morbidity and mortality in 100 consecutive patients, almost all admitted within one week after a proven subarachnoid hemorrhage, were investigated in a prospective study. An aneurysm was demonstrated by angiography or autopsy in 76 patients, while in 24 patients the cerebral panangiography was normal. 13.1% of the patients with a proven aneurysm died within 48 hours of the hemorrhage as a consequence of a massive initial bleeding, 23.6% died from preoperative rebleeding or brain infarcts due to cerebral vascular spasms, 2.6% from direct operative complications and 2.6% from brain infarcts due to postoperative cerebral vascular spasms. The total case mortality was 41.9%. This high mortality was due to massive initial hemorrhage, early rebleedings and in particular to pre- and postoperative brain infarcts due to cerebral vascular spasms. The prognosis can only be improved significantly by early diagnosis and admission and effective prophylaxis and therapy of the vasospasm, but not by further perfecting of surgical technique.


The effect of Ting point (tendinomuscular meridians) electroacupuncture on thermal pain: a model for studying the neuronal mechanism of acupuncture analgesia.
The aim of this study was to characterize the role of Ting points (TP) in acute pain management and its potential use in functional imaging studies by quantitatively assessing: (1) the change in peripheral thermal thresholds before and after the electroacupuncture (EA); and (2) the corresponding behavioral feedback of thermal pain stimulation and the de qi sensation of EA. The study design was prospective. Healthy subjects were recruited for the study at the University of California, San Diego Medical Center. Thirteen (13) healthy subjects were studied. Baseline thermal thresholds (cold and warm sensations and cold and hot pain) were measured at premarked testing sites along the medial aspects of bilateral lower extremities. Five (5) seconds of hot pain (HP) was delivered to the testing sites and the corresponding pain visual analog scale (VAS) scores were recorded. Thirty (30) seconds of EA was delivered via the SP1 and LR1 on the left lower extremities at 5 Hz via a 6-V square-wave stimulator. The VAS scores of the HP and de qi sensation (tingling) during the EA were recorded. The thermal thresholds and VAS scores for the HP and de qi were obtained immediately and both 30 and 60 minutes after the EA. Adaptation testing was also carried out to assess the change in thermal thresholds and the VAS scores of HP without EA. The warm thresholds of bilateral medial calves significantly increased (p < 0.01) after 30 seconds of EA stimulation. The HP VAS score reduced significantly at the ipsilateral calf during EA in comparison to preacupuncture and postacupuncture (p < 0.01) measurements. No significant change in thermal thresholds was noted in the adaptation paradigm. EA at the TP has an inhibitory effect on the C-fiber afferents. The analgesic benefit observed is most likely A-delta afferent mediated. Further correlation studies in functional imaging may provide defining data for the observed analgesic mechanism.


Developing assessment of emergency nurse practitioner competence--a pilot study.
This paper reports a study evaluating baseline competence levels among Emergency Nurse Practitioners using an Objective Structured Clinical Examination. The study also aimed to document change in competence over time and following an educational intervention. Emergency Nurse Practitioners form an increasingly established part of the minor injury service workforce in the United Kingdom (UK). At present, there is no national requirement for them to undergo formal preparation, gain a formal qualification, have continuing professional development or revalidate. Given that it is certain that the use of these professionals to deliver unplanned emergency care will increase in the UK, skill maintenance and extension of skills need to be considered in order to ensure the continued delivery of a quality service. Objective Structured Clinical Examinations aim to test a wide range of knowledge and skills in an objective fashion and can be developed to serve a number of educational requirements. A competence assessment was planned using an Objective Structured Clinical Examination. It was developed following questionnaire consultation with Emergency Nurse Practitioners and also in conjunction with Emergency Department specialists and experts in medical education. The Objective Structured Clinical Examination was piloted with Emergency Department junior doctors. Baseline Objective Structured Clinical Examination assessment was conducted in 2001. An educational intervention followed and then a further Objective Structured Clinical Examination assessment took place in 2002. A total of 17 of 20 Emergency Nurse Practitioners consented to involvement in the study. Comparison of their performance showed a statistically significant overall improvement in performance between the two assessment periods (P < 0.05). Participant feedback on the experience of undertaking an Objective Structured Clinical Examination assessment was very positive. The assessment process worked well, and provides a framework for competence assessment that can be compared over time, between practitioners and between departments.


A role for human Sp alpha as a pattern recognition receptor.
Human Sp alpha is a soluble protein belonging to group B of the scavenger receptor cysteine-rich (SRCR) superfamily for which little functional information is available. It is expressed by macrophages present in lymphoid tissues (spleen, lymph node, thymus, and bone marrow), and it binds to myelomonocytic and lymphoid cells, which suggests that it may play an important role in the regulation of the innate and adaptive immune systems. In the present study we show that recombinant human Sp alpha (rSp alpha) binds to the surface of several gram-positive and gram-negative bacterial strains. Competition studies indicated that such binding is mediated by the recognition of lipoteichoic acid (LTA) and lipopolysaccharide (LPS), respectively, through nonoverlapping sites on the Sp alpha molecule. The most conserved part of LPS (2-keto-3-deoxyoctulosonic acid and lipid A) was shown to be involved in the recognition by Sp alpha. Bacterial binding studies using the SRCR domain 1 of Sp alpha showed that this domain retains both the LPS and LTA binding activities, indicating that both bacterial interacting sites are retained in a single SRCR domain. Furthermore, rSp alpha induced aggregation of gram-positive and gram-negative bacteria strains. On the other hand, rSp alpha inhibited tumor necrosis factor-alpha secretion by human monocytes stimulated with LPS or LTA. Binding of Sp alpha to conserved components of bacterial surfaces and modulation of the monocyte response indicate that this molecule is an active constituent of the innate immune response of the host.


Pulsed-Mode Metalorganic Vapor-Phase Epitaxy of GaN on Graphene-Coated c-Sapphire for Freestanding GaN Thin Films.
We report the growth of high-quality GaN epitaxial thin films on graphene-coated c-sapphire substrates using pulsed-mode metalorganic vapor-phase epitaxy, together with the fabrication of freestanding GaN films by simple mechanical exfoliation for transferable light-emitting diodes (LEDs). High-quality GaN films grown on the graphene-coated sapphire substrates were easily lifted off by using thermal release tape and transferred onto foreign substrates. Furthermore, we revealed that the pulsed operation of ammonia flow during GaN growth was a critical factor for the fabrication of high-quality freestanding GaN films. These films, exhibiting excellent single crystallinity, were utilized to fabricate transferable GaN LEDs by heteroepitaxially growing InxGa1-xN/GaN multiple quantum wells and a p-GaN layer on the GaN films, showing their potential application in advanced optoelectronic devices.


AIDS-related malignancies: state of the art and therapeutic challenges.
Despite the impact of combination antiretroviral therapy (cART) on HIV-related mortality, malignancy remains an important cause of death in the current era. Although the advent of cART has resulted in reductions in the incidence of Kaposi's sarcoma and non-Hodgkin's lymphoma, non-AIDS-defining malignancies present an increased risk for HIV-infected patients, characterized by some common clinical features, generally with a more aggressive behavior and a more advanced disease at diagnosis, which is responsible for poorer patient outcomes. Specific therapeutic recommendations are lacking for these new nonopportunistic malignancies, such as Hodgkin's lymphoma, anal cancer, lung cancer, hepatocarcinoma, and many others. Antiretroviral agents have a propensity for causing drug interactions as a result of their ability to either inhibit or induce the cytochrome P450 (CYP) enzyme system. Because many antineoplastic drugs are also metabolized by the CYP system, coadministration with cART could result in either drug accumulation with increased toxicity, or decreased efficacy of one or both classes of drugs. Further research delineating the combined safety and pharmacokinetics of antiretrovirals and antineoplastic therapy is necessary. Special considerations of these AIDS-related and non-AIDS-related malignancies and their clinical and therapeutic aspects constitute the subject of this review.


Introducing DEPDOSE, a Tool to Calculate Dose Coefficients to Members of the Public for Radioactive Aerosols.
This paper presents DEPDOSE, an open-source computer application that combines the KDEP respiratory tract deposition fractions for inhaled aerosols with DC_PAK committed equivalent dose coefficients for a unit deposition in each region of the respiratory tract. DEPDOSE allows the user to rapidly produce tables of dose coefficients for workers and members of the public inhaling precisely defined, user-specified aerosols using the ICRP Publication 60 methodology. Combined with a plume dispersion modeling system, such as the Quick Urban & Industrial Complex (QUIC) Dispersion Modeling System, this makes it possible to predict radiation doses downstream from an accidental or intentional release of radioactive materials. For this work, a radioactive plume was calculated to members of the public downstream from a dirty bomb in Chicago. DEPDOSE is published under an open source license, and can be downloaded at https://github.com/lanl/DEPDOSE .


Antibiotic susceptibility among Staphylococcus epidermidis isolated from prosthetic joint infections with special focus on rifampicin and variability of the rpoB gene.
Staphylococcus epidermidis is the most important pathogen in infections related to implanted foreign materials, especially prosthetic joint infections (PJIs). The aim of this study was to investigate the antimicrobial activities of 16 antibiotics against S. epidermidis isolated from PJIs, with special focus on rifampicin and rpoB variability. Ninety-one per cent of the isolates were multiresistant (i.e. resistant to members of more than three classes of antibiotics). Thirty-nine per cent were resistant to rifampicin, associated with one or two single-nucleotide polymorphisms (SNPs) in rpoB. Using IsoSensitest agar with supplements, 61% were resistant to oxacillin, and using Mueller-Hinton II agar with supplement, 84% were resistant. Using the Etest, 58% were resistant to cefoxitin, and using the disk diffusion test, 91% were resistant. The mecA gene was detected in 85% of the isolates. Regarding recently available antibiotics, all isolates were susceptible to tigecycline and linezolid, and 97% were susceptible to daptomycin. In addition, two novel antibiotics, dalbavancin and ceftobiprole, were tested, although not yet available for routine use. The MIC(50) and MIC(90) values of these novel antibiotics were 0.032 and 0.047 mg/L and 0.5 and 1.5 mg/L, respectively. Among the other antibiotics, the rates of resistance varied between 0% (vancomycin) and 82% (trimethoprim-sulphamethoxazole). S. epidermidis strains causing PJIs often show multiresistance, including resistance to rifampicin, which is mainly caused by one or two SNPs. Some of the newer antimicrobial agents may provide alternatives for monotherapy or combination therapy with rifampicin. Detection of mecA is necessary before initiating treatment of infections due to S. epidermidis when it displays intermediate susceptibility to cefoxitin.


Disease resistance of pacu Piaractus mesopotamicus (Holmberg, 1887) fed with β-glucan.
Effects of β-glucan on innate immune responses and survival were studied in pacu experimentally infected with Aeromonas hydrophila. Fish fed diets containing 0, 0.1% and 1% β-glucan were injected with A. hydrophila. β-glucan enhanced fish survival in both treated groups (26.7% and 21.2% of the control, respectively). Leukocyte respiratory burst and alternative complement pathway activities were elevated after bacterial challenge regardless the β-glucan concentration. Lysozyme activity was higher after infection and showed a gradual increase as β-glucan concentration increased. A significant elevation in WBC count was observed either after bacterial challenge or by influence of β-glucan separately. The same response was observed in the number of thrombocytes, lymphocytes, eosinophils, LG-PAS positive cell and monocytes. It can be concluded that feeding pacu with β-glucan can increase protection against A. hydrophila, due to changes in non-specific immune responses.


Computer-assisted quantitation of choroidal neovascularization for clinical trials.
To develop a computer-assisted method for the quantitation of choroidal neovascularization (CNV) for the support of clinical trials. Fluorescein angiographic images were selected from 5 patients enrolled in a clinical trial for which three follow-up visits were available. Thirty- and 600-second images were digitized at 1000 dots/in and registered (aligned) with polynomial warping algorithms. Custom-developed software allowed for coarse, automated identification of CNV. An easy-to-use graphical user interface facilitated supervision and refinement of the lesion boundaries by a skilled reader based on standard stereoscopic viewing of the fluorescein angiography study. Capabilities for boundary delineation in both early and late phases, and animation to allow for image correlation and evaluation of temporal changes in fluorescence of spatially corresponding pixels, were included. Two metrics for CNV characterization were generated. First, the lesion area based on the lesion boundaries was identified after supervision. Second, an integrated lesion intensity (ILI) reflecting the integrated, normalized lesion hyperfluorescence was calculated. Area and ILI measures were calculated for each of 5 patients for three or more visits. Facile supervision based on the stereoscopic angiogram permitted arbitrarily close concordance with CNV identification using standard methods. Changes in area and ILI measurements between visits correlated closely with clinically observed changes in each case. Interactive image processing permits efficient, accurate, computer-assisted CNV quantitation that may be useful for the support of clinical trials and preclinical studies.


Playing the odds: lung cancer surveillance after curative surgery.
After 'curative' resection, many patients are still at risk for further lung cancer, either as a recurrence or a new metachronous primary. In theory, close follow-up should improve survival by catching relapse early - but in reality, many experts feel that surveillance for recurrence is of uncertain value. In this article, we explore the reasons behind the controversy, what the current guidelines recommend, and what future solutions are in development that may ultimately resolve this debate. Although postoperative surveillance for a new lung cancer may impart a survival advantage, this benefit does not appear to extend to the phenomenon of recurrence. Nevertheless, close radiographic follow-up after curative resection is still recommended by most professional societies, with more frequent scanning in the first 2 years, and then annual screening thereafter. Given the radiation risk, however, low-dose and minimal-dose computed tomography options are under investigation, as well as timing scans around expected peaks of recurrence rather than a set schedule. Applying the same surveillance algorithm to all lung cancer patients after curative resection may not be cost-effective or reasonable, especially if there is no demonstrable mortality benefit. Therefore, future research should focus on finding safer nonradiographic screening options, such as blood or breath biomarkers, or developing nomograms for predicting which patients will relapse and require closer follow-up. Ultimately, however, better tools for surveillance may be moot until we develop better treatment options for lung cancer recurrence.


The role of EDTA in lead transport and accumulation by indian mustard
Indian mustard (Brassica juncea) plants exposed to Pb and EDTA in hydroponic solution were able to accumulate up to 55 mmol kg-1 Pb in dry shoot tissue (1.1% [w/w]). This represents a 75-fold concentration of Pb in shoot tissue over that in solution. A threshold concentration of EDTA (0.25 mm) was found to be required to stimulate this dramatic accumulation of both Pb and EDTA in shoots. Below this threshold concentration, EDTA also accumulated in shoots but at a reduced rate. Direct measurement of a complex of Pb and EDTA (Pb-EDTA) in xylem exudate of Indian mustard confirmed that the majority of Pb in these plants is transported in coordination with EDTA. The accumulation of EDTA in shoot tissue was also observed to be directly correlated with the accumulation of Pb. Exposure of Indian mustard to high concentrations of Pb and EDTA caused reductions in both the transpiration rate and the shoot water content. The onset of these symptoms was correlated with the presence of free protonated EDTA (H-EDTA) in the hydroponic solution, suggesting that free H-EDTA is more phytotoxic than Pb-EDTA. These studies clearly demonstrate that coordination of Pb transport by EDTA enhances the mobility within the plants of this otherwise insoluble metal ion, allowing plants to accumulate high concentrations of Pb in shoots. The finding that both H-EDTA and Pb-EDTA are mobile within plants also has important implications for the use of metal chelates in plant nutritional research.


Comparative study of scientific publications in orthopedics journals originating from USA, Japan and China (2000-2012).
To compare orthopedics publications from USA, Japan and China. Scientific papers belong to ''Orthopedics'' category of Science Citation Index Expanded subject categories were retrieved from the "PubMed'' and ''Web of Knowledge'' online databases. In the field of orthopedics, the annual number increased significantly from 2000 to 2012 in the three countries (p<0.001). The share of articles increased significantly in China, but decreased significantly in Japan and USA (p<0.05). In 2012, USA contributed 35.3% of the total world output in orthopedics field and ranked 1st; Japan contributed 5.9% and ranked 4th; China contributed 5.2% and ranked 5th. Publications from USA had the highest accumulated IFs and the highest total citations of articles (USA > Japan > China, p<0.001). Average IF from USA was much higher than Japan and China (p<0.001). USA published the most articles in the top ten orthopedics journals (USA (14355) > Japan (1702) > China (487), p<0.01). Although China has undergone significant increase in annual number and percentage of scientific publication in orthopedics journals, it still lags far behind USA and Japan in the field of orthopedics in terms of quantity and quality.


Protective Effects of L-902,688, a Prostanoid EP4 Receptor Agonist, against Acute Blood-Brain Barrier Damage in Experimental Ischemic Stroke.
Ischemic stroke occurs when a clot forms in the brain vasculature that starves downstream tissue of oxygen and nutrients resulting in cell death. The tissue immediately downstream of the blockage, the core, dies within minutes, but the surrounding tissue, the penumbra is potentially salvageable. Prostaglandin E2 binds to four different G-protein coupled membrane receptors EP1-EP4 mediating different and sometimes opposing responses. Pharmacological activation of the EP4 receptor has already been established as neuroprotective in stroke, but the mechanism(s) of protection are not well-characterized. In this study, we hypothesized that EP4 receptor activation reduces ischemic brain injury by reducing matrix metalloproteinase (MMP)-3/-9 production and blood-brain barrier (BBB) damage. Rats underwent transient ischemic stroke for 90 min. Animals received an intravenous injection of either the vehicle or L-902,688, a highly specific EP4 agonist, at the onset of reperfusion. Brain tissue was harvested at 24 h. We established a dose-response curve and used the optimal dose that resulted in the greatest infarct reduction to analyze BBB integrity compared to vehicle-treated rats. The presence of IgG, a blood protein, in the brain parenchyma is a marker of BBB damage, and L-902,688 (1 mg/kg; i.v.) dramatically reduced IgG extravasation (P < 0.05). Consistent with these data, we assessed zona occludens-1 and occludin, tight junction proteins integral to the maintenance of the BBB, and found reduced degradation with L-902,688 administration. With immunoblotting, qRT-PCR, and/or a fluorescence resonance energy transfer (FRET)-based activity assay, we next measured MMP-3/-9 since they are key effectors of BBB breakdown in stroke. In the cerebral cortex, not only was MMP-3 activity significantly decreased (P < 0.05), but L-902,688 treatment also reduced MMP-9 mRNA, protein, and enzymatic activity (P < 0.001). In addition, post-ischemic administration of the EP4 agonist significantly reduced pro-inflammatory cytokines IL-1β (P < 0.05) and IL-6 (P < 0.01) in the ischemic cerebral cortex. Most importantly, one injection of L-902,688 (1 mg/kg; i.v) at the onset of reperfusion significantly reduces neurological deficits up to 3 weeks later (P < 0.05). Our data show for the first time that pharmacological activation of EP4 with L-902,688 is neuroprotective in ischemic stroke by reducing MMP-3/-9 and BBB damage.