Direct participation of Sam68, the 68-kilodalton Src-associated protein in mitosis, in the CRM1-mediated Rev nuclear export pathway.
Human immunodeficiency virus type 1 (HIV-1) replication requires efficient nuclear export of incompletely spliced and unspliced HIV-1 mRNA transcripts, which is achieved by Rev expression at an early stage of the viral life cycle. We have recently shown that expression of Sam68, the 68-kDa Src-associated protein in mitosis, is able to alleviate Rev function block in astrocytes by promoting Rev nuclear export. In the present study, we utilized an antisense RNA expression strategy to down-modulate constitutive Sam68 expression and examined its effect on Rev function, HIV-1 gene expression, and viral replication. These results showed that down-modulation of constitutive Sam68 expression markedly inhibited HIV-1 production in 293T cells and viral replication in T lymphocytes such as Jurkat and CEM cells, as well as human peripheral blood mononuclear cells (PBMCs). Rev-dependent in trans complementation and reporter gene assays further demonstrated that inhibition of HIV-1 gene expression by Sam68 down-modulation was due to impeded Rev activity. Moreover, digital fluorescence microscopic imaging revealed that down-modulation of Sam68 expression caused exclusive nuclear retention and colocalization of both Rev and CRM1. Taken together, these data suggest that adequate Sam68 expression is required for Rev function and, thereby, for HIV-1 gene expression and viral replication, and they support the notion that Sam68 is directly involved in the CRM1-mediated Rev nuclear export pathway.


Historical perspective on surfactant therapy: Transforming hyaline membrane disease to respiratory distress syndrome.
Lung surfactant is the first drug so far designed for the special needs of the newborn. In 1929, Von Neergard described lung hysteresis and proposed the role of surface forces. In 1955-1956, Pattle and Clements found direct evidence of lung surfactant. In 1959, Avery discovered that the airway's lining material was not surface-active in hyaline membrane disease (HMD). Patrick Bouvier Kennedy's death, among half-million other HMD-victims in 1963, stimulated surfactant research. The first large surfactant treatment trial failed in 1967, but by 1973, prediction of respiratory distress syndrome using surfactant biomarkers and promising data on experimental surfactant treatment were reported. After experimental studies on surfactant treatment provided insight in lung surfactant biology and pharmacodynamics, the first trials of surfactant treatment conducted in the 1980s showed a striking amelioration of severe HMD and its related deaths. In the 1990s, the first synthetic and natural surfactants were accepted for treatment of infants. Meta-analyses and further discoveries confirmed and extended these results. Surfactant development continues as a success-story of neonatal research.


Lidar measurements of solid rocket propellant fire particle plumes.
This paper presents the first, to our knowledge, direct measurement of aerosol produced by an aluminized solid rocket propellant (SRP) fire on the ground. Such fires produce aluminum oxide particles small enough to loft high into the atmosphere and disperse over a wide area. These results can be applied to spacecraft launchpad accidents that expose spacecraft to such fires; during these fires, there is concern that some of the plutonium from the spacecraft power system will be carried with the aerosols. Accident-related lofting of this material would be the net result of many contributing processes that are currently being evaluated. To resolve the complexity of fire processes, a self-consistent model of the ground-level and upper-level parts of the plume was determined by merging ground-level optical measurements of the fire with lidar measurements of the aerosol plume at height during a series of SRP fire tests that simulated propellant fire accident scenarios. On the basis of the measurements and model results, the Johns Hopkins University Applied Physics Laboratory (JHU/APL) team was able to estimate the amount of aluminum oxide (alumina) lofted into the atmosphere above the fire. The quantification of this ratio is critical for a complete understanding of accident scenarios, because contaminants are transported through the plume. This paper provides an estimate for the mass of alumina lofted into the air.


Tumor cell entry into the lymph node is controlled by CCL1 chemokine expressed by lymph node lymphatic sinuses.
Lymphatic vessels are thought to contribute to metastasis primarily by serving as a transportation system. It is widely believed that tumor cells enter lymph nodes passively by the flow of lymph. We demonstrate that lymph node lymphatic sinuses control tumor cell entry into the lymph node, which requires active tumor cell migration. In human and mouse tissues, CCL1 protein is detected in lymph node lymphatic sinuses but not in the peripheral lymphatics. CCR8, the receptor for CCL1, is strongly expressed by human malignant melanoma. Tumor cell migration to lymphatic endothelial cells (LECs) in vitro is inhibited by blocking CCR8 or CCL1, and recombinant CCL1 promotes migration of CCR8(+) tumor cells. The proinflammatory mediators TNF, IL-1β, and LPS increase CCL1 production by LECs and tumor cell migration to LECs. In a mouse model, blocking CCR8 with the soluble antagonist or knockdown with shRNA significantly decreased lymph node metastasis. Notably, inhibition of CCR8 led to the arrest of tumor cells in the collecting lymphatic vessels at the junction with the lymph node subcapsular sinus. These data identify a novel function for CCL1-CCR8 in metastasis and lymph node LECs as a critical checkpoint for the entry of metastases into the lymph nodes.


Divergent pharmacological effects of three calmodulin antagonists, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), chlorpromazine and calmidazolium, on isometric tension development and myosin light chain phosphorylation in intact bovine tracheal smooth muscle.
To investigate the usefulness of calmodulin antagonists in intact cell systems, effects of three calmodulin antagonists, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), chlorpromazine (CPZ) and calmidazolium on isometric tension development and myosin light chain (P-LC) phosphorylation in bovine tracheal smooth muscle strips were compared to inhibition of purified myosin light chain kinase activity. These antagonists inhibited the Ca++-calmodulin-induced activation of myosin light chain kinase in a concentration-dependent manner, with IC50 values of 1.0 (calmidazolium), 25 (W-7) and 65 microM (CPZ), respectively. Inhibitory effects of these antagonists were abolished with increasing concentrations of calmodulin. However, when these antagonists were used in intact smooth muscle strips, the gradation of potencies did not parallel the anticalmodulin activities. W-7 (100 microM) exhibited a similar extent of antagonism between the contractile responses to carbachol and KCl. The increase in P-LC phosphate content in response to 1-min stimulation with 10(-5) M carbachol was inhibited by W-7. CPZ exhibited an unexpectedly potent antagonism on carbachol-induced isometric tension development and P-LC phosphorylation. Atropine showed an antagonism similar to CPZ. CPZ and verapamil had similar antagonistic effects on KCI-induced contractions. Calmidazolium (50 microM) produced no significant inhibition on carbachol-induced isometric tension development and P-LC phosphorylation in intact smooth muscle strips. It may be concluded that 1) W-7 antagonizes the smooth muscle contraction through the inhibition of the initial increase in the P-LC phosphorylation; 2) CPZ produces effects other than calmodulin antagonism; and 3) calmidazolium is not effective in intact smooth muscle strips.(ABSTRACT TRUNCATED AT 250 WORDS)


Optical compressive change and motion detection.
Localization information of moving and changing objects, as commonly extracted from video sequences, is typically very sparse with respect to the full data frames, thus fulfilling one of the basic conditions of compressive sensing theory. Motivated by this observation, we developed an optical compressive change and motion-sensing technique that detects the location of moving objects by using a significantly fewer samples than conventionally taken. We present examples of motion detection and motion tracking with over two orders of magnitude fewer samples than required with conventional systems.


Biochemical basis of permethrin resistance in Anopheles arabiensis from Lower Moshi, north-eastern Tanzania.
Development of resistance to different classes of insecticides is a potential threat to malaria control. With the increasing coverage of long-lasting insecticide-treated nets in Tanzania, the continued monitoring of resistance in vector populations is crucial. It may facilitate the development of novel strategies to prevent or minimize the spread of resistance. In this study, metabolic-based mechanisms conferring permethrin (pyrethroid) resistance were investigated in Anopheles arabiensis of Lower Moshi, Kilimanjaro region of north-eastern Tanzania. WHO susceptibility test kits were used to detect resistance to permethrin in An. arabiensis. The levels and mechanisms of permethrin resistance were determined using CDC bottle bioassays and microplate (biochemical) assays. In bottle bioassays, piperonyl butoxide (PBO) and s,s,s-tributyl phosphorotrithioate (DEF) were used as synergists to inhibit mixed function oxidases and non-specific esterases respectively. Biochemical assays were carried out in individual mosquitoes to detect any increase in the activity of enzymes typically involved in insecticide metabolism (mixed function oxidases, alpha- and beta-esterases). Anopheles arabiensis from the study area was found to be partially resistant to permethrin, giving only 87% mortality in WHO test kits. Resistance ratios at KT50 and KT95 were 4.0 and 4.3 respectively. The permethrin resistance was partially synergized by DEF and by PBO when these were mixed with permethrin in bottle bioassays and was fully synergized when DEF and PBO were used together. The levels of oxidase and beta-esterase activity were significantly higher in An. arabiensis from Lower Moshi than in the laboratory susceptible strain. There was no difference in alpha-esterase activity between the two strains. Elevated levels of mixed function oxidases and beta-esterases play a role in detoxification of permethrin in the resistant An. arabiensis population of Lower Moshi.


Remote sensing for assessing the zone of benefit where deep drains improve productivity of land affected by shallow saline groundwater.
The installation of deep drains is an engineering approach to remediate land salinised by the influence of shallow groundwater. It is a costly treatment and its economic viability is, in part, dependent on the lateral extent to which the drain increases biological productivity by lowering water tables and soil salinity (referred to as the drains' zone of benefit). Such zones may be determined by assessing the biological productivity response of adjacent vegetation over time. We tested a multi-temporal satellite remote sensing method to analyse temporal and spatial changes in vegetation condition surrounding deep drainage sites at five locations in the Western Australian wheatbelt affected by dryland salinity-Morawa, Pithara, Beacon, Narembeen and Dumbleyung. Vegetation condition as a surrogate for biological productivity was assessed by Normalised Difference Vegetation Index (NDVI) during the peak growing season. Analysis was at the site scale within a 1000 m buffer zone from the drains. There was clear evidence of NDVI increasing with elevation, slope and distance from the drain. After accounting for elevation, slope and distance from the drain, there was a significant increase in NDVI across the five locations after installation of deep drains. Changes in NDVI after drainage were broadly consistent with measured changes at each site in groundwater levels after installation of the deep drains. However, this study assessed the lateral extent of benefit for biological productivity and gave a measure of the area of benefit along the entire length of the drain. The method demonstrated the utility of spring NDVI images for rapid and relatively simple assessment of the change in site condition after implementation of drainage, but approaches for further improvement of the procedure were identified.


Catalytic Cycle of Haloalkane Dehalogenases Toward Unnatural Substrates Explored by Computational Modeling.
The anthropogenic toxic compound 1,2,3-trichloropropane is poorly degradable by natural enzymes. We have previously constructed the haloalkane dehalogenase DhaA31 by focused directed evolution ( Pavlova, M. et al. Nat. Chem. Biol. 2009 , 5 , 727 - 733 ), which is 32 times more active than the wild-type enzyme and is currently the most active variant known against that substrate. Recent evidence has shown that the structural basis responsible for the higher activity of DhaA31 was poorly understood. Here we have undertaken a comprehensive computational study of the main steps involved in the biocatalytic hydrolysis of 1,2,3-trichloropropane to decipher the structural basis for such enhancements. Using molecular dynamics and quantum mechanics approaches we have surveyed (i) the substrate binding, (ii) the formation of the reactive complex, (iii) the chemical step, and (iv) the release of the products. We showed that the binding of the substrate and its transport through the molecular tunnel to the active site is a relatively fast process. The cleavage of the carbon-halogen bond was previously identified as the rate-limiting step in the wild-type. Here we demonstrate that this step was enhanced in DhaA31 due to a significantly higher number of reactive configurations of the substrate and a decrease of the energy barrier to the SN2 reaction. C176Y and V245F were identified as the key mutations responsible for most of those improvements. The release of the alcohol product was found to be the rate-limiting step in DhaA31 primarily due to the C176Y mutation. Mutational dissection of DhaA31 and kinetic analysis of the intermediate mutants confirmed the theoretical observations. Overall, our comprehensive computational approach has unveiled mechanistic details of the catalytic cycle which will enable a balanced design of more efficient enzymes. This approach is applicable to deepen the biochemical knowledge of a large number of other systems and may contribute to robust strategies in the development of new biocatalysts.


Amnion epithelial cells - a novel therapy for ischemic stroke?
Stroke is a leading cause of death and disability and new therapies are desperately needed. Given the complex nature of ischemic brain injury, it has been postulated that cell-based therapies may be useful. However, cell resources, invasive extraction procedures, immunological rejection, tumorigenesis and ethical challenges make it unlikely that many stem cell types could serve as a practical source for therapy. By contrast, these issues do not pertain to human amnion epithelial cells (hAECs), which are placenta-derived stem cells. We recently assessed the effects of systemically delivered hAECs on stroke outcome using four animal models of stroke. We demonstrated that when injected intravenously after ischemia onset, hAECs migrate preferentially to the spleen and injured brain to limit apoptosis and inflammation, and attenuate early brain infiltration of immune cells, progression of infarction and systemic immunosuppression and to ultimately ameliorate functional deficits. When administration of hAECs is delayed by 1-3 days post-stroke, long-term functional recovery can still be enhanced in young and aged mice of either sex. Moreover, our proof-of-principle findings suggest that hAECs are effective at limiting post-stroke infarct development in non-human primates. Overall, the results suggest that hAECs could be a viable clinical stroke therapy.


Carbohydrate specificity of sea urchin sperm bindin: a cell surface lectin mediating sperm-egg adhesion.
We have examined the carbohydrate specificity of bindin, a sperm protein responsible for the adhesion of sea urchin sperm to eggs, by investigating the interaction of a number of polysaccharides and glycoconjugates with isolated bindin. Several of these polysaccharides inhibit the agglutination of eggs by bindin particles. An egg surface polysaccharide was found to be the most potent inhibitor of bindin-mediated egg agglutination. Fucoidin, a sulfated fucose heteropolysaccharide, was the next most potent inhibitor, followed by the egg jelly fucan, a sulfated fucose homopolysaccharide, and xylan, a beta(1 leads to 4) linked xylose polysaccharide. A wide variety of other polysaccharides and glycoconjugates were found to have no effect on egg agglutination. We also report that isolated bindin has a soluble lectinlike activity which is assayed by agglutination of erythrocytes. The bindin lectin activity is inhibited by the same polysaccharides that inhibit egg agglutination by particulate bindin. This suggests that the egg adhesion activity of bindin is directly related to its lectin activity. We have established that fucoidin binds specifically to bindin particles with a high apparent affinity (Kd = 5.5 X 10(-8) M). The other polysaccharides that inhibit egg agglutination also inhibit the binding of 125I-fucoidin to bindin particles, suggesting that they compete for the same site on bindin. The observation that polysaccharides of different composition and linkage type interact with bindin suggests that the critical structural features required for binding may reside at a higher level of organization. Together, these findings strengthen the hypothesis that sperm-egg adhesion in sea urchins is mediated by a lectin-polysaccharide type of interaction.


The Gynecologist Has a Unique Role in Providing Oncofertility Care to Young Cancer Patients.
Facing a cancer diagnosis at any age is devastating. However, young cancer patients have the added burden that life-preserving cancer treatments, including surgery, chemotherapy, and radiotherapy, may compromise their future fertility. The possibility of reproductive dysfunction as a consequence of cancer treatment has a negative impact on the quality of life of cancer survivors. The field of oncofertility, which merges the clinical specialties of oncology and reproductive endocrinology, was developed to explore and expand fertility preservation options and to better manage the reproductive status of cancer patients. Fertility preservation for females has proved to be a particular challenge because mature female gametes are rare and difficult to acquire. The purpose of this article is to provide the gynecologist with a comprehensive overview of how cancer treatments affect the female reproductive axis, delineate the diverse fertility preservation options that are currently available or being developed for young women, and describe current measures of ovarian reserve that can be used pre- and post-cancer treatment. As a primary care provider, the gynecologist will likely interact with patients throughout the cancer care continuum. Thus, the gynecologist is in a unique position to join the oncofertility team in providing young cancer patients with up-to-date fertility preservation information and referrals to specialists.


A psychometric cut-point to separate emergently suicidal depressed patients from stable depressed outpatients.
The design of safe clinical trials targeting suicidal ideation requires operational definitions of what degree of suicidal ideation is too excessive to allow safe participation. We examined the Scale for Suicide Ideation (SSI) to develop a psychometric cut-point that would identify patients having a suicidal emergency. The Emergency Department (ED) and the out-patient clinic of a university hospital. We used the SSI to contrast 23 stable, depressed adult out-patients versus 11 depressed adult ED patients awaiting psychiatric admission for a suicidal emergency. The performance of the SSI was examined with nominal logistic regression. ED patients were older than out-patients (P<0.001), with proportionally more men (P<0.05), and were more ethnically diverse than the outpatients (P<0.005). Compared to out-patients, ED patients were more depressed (Patient Health Questionnaire-9 score 23.1±3.8 vs. 11.7±7.3, P<0.005) and reported a greater degree of suicidal ideation (SSI scores 25.7±7.3 vs. 4.2±8.4, P<0.0001). Nominal logistic regression for the univariate model of SSI score and group yielded a score of 16 (P<0.0001) as the best cut-point in separating groups, with a corresponding Receiver Operating Characteristic Area Under the Curve = 0.94. Of 34 patients in the total sample, only two were misclassified by SSI score = 16, with both of these being false positive for ED status. Thus, the sensitivity of the cut-point was 100% with specificity of 91%. When the model was expanded to include SSI along with age, gender, ethnicity, sedative-hypnotic use, and over-the-counter use, only SSI score remained significant as a predictor. A SSI score ≥16 may be useful as an exclusion criterion for out-patients in depression clinical trials.


Umbilical cord endothelial cells expressing large T antigen: comparison with primary cultures and effect of cell age.
A number of human endothelial cell lines from umbilical cord cells (HUVECs) have been generated by transfection with SV40 large T and small t antigen sequences. Comparison of these lines with primary cultures of HUVECs has been carried out by monitoring the expression of a number of endothelial cell markers with specific regard to cell age. The secreted levels of the protein plasminogen activator inhibitor (PAI) was found to be significantly reduced in SV40-transfected cells when compared to untransfected controls. Tissue plasminogen activator (tPA) and urokinase (uPA) levels were unchanged. As cells entered crisis, there was a rapid and significant increase in the levels of tPA, uPA, and PAl and this was observed for all clones screened. The endothelial cell marker von Willebrand Factor (vWF) was found intracellularly and was also secreted into the medium. The levels were not altered between transfected and untransfected cells. Angiotensin converting enzyme (ACE) activity was maintained in cell lines at levels found in nonimmortalized HUVECs. Both isoforms (alpha and beta) of IL-1 (interleukin-1) increased as cells approached crisis, and the presence of these cytokines may be responsible for the increased levels of tPA, PAI, and uPA. With one exception, the ability of the transfected cells to produce prostacyclin (PGI2) was lost by all clones.


pTransgenesis: a cross-species, modular transgenesis resource.
As studies aim increasingly to understand key, evolutionarily conserved properties of biological systems, the ability to move transgenesis experiments efficiently between organisms becomes essential. DNA constructions used in transgenesis usually contain four elements, including sequences that facilitate transgene genome integration, a selectable marker and promoter elements driving a coding gene. Linking these four elements in a DNA construction, however, can be a rate-limiting step in the design and creation of transgenic organisms. In order to expedite the construction process and to facilitate cross-species collaborations, we have incorporated the four common elements of transgenesis into a modular, recombination-based cloning system called pTransgenesis. Within this framework, we created a library of useful coding sequences, such as various fluorescent protein, Gal4, Cre-recombinase and dominant-negative receptor constructs, which are designed to be coupled to modular, species-compatible selectable markers, promoters and transgenesis facilitation sequences. Using pTransgenesis in Xenopus, we demonstrate Gal4-UAS binary expression, Cre-loxP-mediated fate-mapping and the establishment of novel, tissue-specific transgenic lines. Importantly, we show that the pTransgenesis resource is also compatible with transgenesis in Drosophila, zebrafish and mammalian cell models. Thus, the pTransgenesis resource fosters a cross-model standardization of commonly used transgenesis elements, streamlines DNA construct creation and facilitates collaboration between researchers working on different model organisms.


Cocoa procyanidins modulate transcriptional pathways linked to inflammation and metabolism in human dendritic cells.
Foods rich in polyphenols such as procyanidins (PC) have been proposed to have anti-inflammatory properties, and we have previously reported inhibition of lipopolysaccharide (LPS)-induced inflammatory cytokine secretion in human dendritic cells (DCs) by PC derived from cocoa. To explore the mechanistic basis of this inhibition, here we conducted transcriptomic analysis on DCs cultured with either LPS or LPS combined with oligomeric cocoa PC. Procyanidins suppressed a number of genes encoding cytokines and chemokines such as CXCL1, but also genes involved in the cGMP pathway such as GUCY1A3 (encoding guanylate cyclase soluble subunit alpha-3). Upregulated genes were involved in diverse metabolic pathways, but notably two of the four most upregulated genes (NMB, encoding neuromedin B and ADCY3, encoding adenyl cyclase type 3) were involved in the cAMP signalling pathway. Gene-set enrichment analysis demonstrated that upregulated gene pathways were primarily involved in nutrient transport, carbohydrate metabolism and lysosome function, whereas down-regulated gene pathways involved cell cycle, signal transduction and gene transcription, as well as immune function. qPCR analysis verified differential expression of GUCY1A3, ADCY3, NMB as well as a number of other genes, and marked suppression of LPS-induced CXCL1 and IL-23 protein secretion was also observed. Thus, our results confirm a marked anti-inflammatory effect of PC in human DCs, which may be related mechanistically to second-messenger function and metabolic activity. Our results provide a foundation to further investigate metabolic pathways altered by PC during intestinal inflammation, and further encourage investigation of the health-promoting potential of PC-rich functional foods.


Dynamics of a microorganism in a sheared viscoelastic liquid.
In this paper, we investigate the dynamics of a model spherical microorganism, called squirmer, suspended in a viscoelastic fluid undergoing unconfined shear flow. The effect of the interplay of shear flow, fluid viscoelasticity, and self-propulsion on the orientational dynamics is addressed. In the limit of weak viscoelasticity, quantified by the Deborah number, an analytical expression for the squirmer angular velocity is derived by means of the generalized reciprocity theorem. Direct finite element simulations are carried out to study the squirmer dynamics at larger Deborah numbers. Our results show that the orientational dynamics of active microorganisms in a sheared viscoelastic fluid greatly differs from that observed in Newtonian suspensions. Fluid viscoelasticity leads to a drift of the particle orientation vector towards the vorticity axis or the flow-gradient plane depending on the Deborah number, the relative weight between the self-propulsion velocity and the flow characteristic velocity, and the type of swimming. Generally, pullers and pushers show an opposite equilibrium orientation. The results reported in the present paper could be helpful in designing devices where separation of microorganisms, based on their self-propulsion mechanism, is obtained.


Anticipatory and compensatory postural adjustments in people with low back pain: a systematic review and meta-analysis.
Despite altered anticipatory (APAs) and compensatory postural adjustments (CPAs) being hypothesized to contribute to the onset and persistence of low back pain (LBP), results from studies comparing people with and without LBP are conflicting. This systematic review aimed to determine whether APAs or CPAs are altered in the presence of acute and chronic LBP. A systematic review of studies was carried out. No patient sample was required. Between group standardized mean differences and 95% confidence intervals for APAs ad CPAs METHODS: A comprehensive search was conducted for articles comparing people with LBP (acute or chronic) to healthy controls for the onset or amplitude of muscle activity, center of pressure (COP), or kinematic responses to expected or unexpected perturbations. Two independent reviewers extracted data and assessed the methodological quality of relevant studies. Differences between people with and without LBP were calculated as standardized mean differences, and included in a meta-analysis if outcomes were homogeneous. Otherwise, a narrative synthesis was conducted. Twenty-seven studies were included, of which the majority examined muscle onsets in response to expected and unexpected perturbations. Only two studies compared people with and without acute LBP, and results for these studies were conflicting. The results show delayed muscle onsets in response to expected and unexpected perturbations for people with chronic LBP when compared with healthy controls. No conclusive evidence for differences between people with and without chronic LBP for COP or kinematic responses. There is currently no convincing evidence of differences between people with and without acute LBP for APAs or CPAs. Conversely, delayed muscle onsets in people with chronic LBP suggest APAs and CPAs are altered in this population. However, the functional relevance of these delayed muscle onsets (eg, COP and kinematics) is unknown.


Effect of age on visual acuity after cataract extraction.
The best Snellen visual acuity in the three months after cataract extraction was found to decline with the age of the patient. The relationship was identical in 111 patients who had extracapsular extraction and a Pearce tripod posterior chamber intraocular lens and in 50 patients who had intracapsular extraction with spectacle correction. Vision after operation varied from a mean value of 6/5 at 50 years to 6/12 at 90 years, a decline of 1 line per 13.4 years. In the intracapsular group, over a mean follow-up period of 14 years, the rate of fall in acuity with increasing age after operation was found to be statistically similar to that of the early postoperative acuity plotted against age for both types of operation. This suggests that the rate of decline with age is unchanged after a prolonged period of aphakia with presumed increased exposure to ultraviolet and blue light. The data were found to be similar to the decline in the neurosensory elements of vision with age measured experimentally in 20 phakic subjects by laser interferometry. This method of assessment of contrast sensitivity threshold effectively bypasses changes in the optical media. The findings indicate that the previously recognised drop in visual acuity with age is not related to changes in the aging crystalline lens and support the view that there is a decline in the neurosensory elements of vision. It is important to recognise this deterioration so that results of surgery or other treatment are adjusted to allow for the age of the patients.


Tinnitus school: an educational approach to tinnitus management based on a stress-reaction tinnitus model.
Stress is a significant factor influencing the clinical course of tinnitus. The auditory system is particularly sensitive to the effects of various stress factors (chemical, oxidative, emotional, etc.). Different stages of reaction (alarm, resistance, exhaustion) lead to different characteristics of tinnitus and to different therapeutic approaches. Individual characteristics of stress reaction may explain different aspects of tinnitus in various patients with different responses to treatment, despite similar audiological and etiological factors. A model based on individual reactions to stress factors (stress-reaction tinnitus model, or SRTM) could explain tinnitus as an alarm signal. In each patient, stressors have to be identified during the alarm phase to prevent an evolution toward the resistance and exhaustion phases. In the exhaustion phase, chronic tinnitus is due to the organization of a paradoxical auditory memory and a pathologically shifted attention to tinnitus. The aim of our study is to describe a therapeutic proposal based on the SRTM by taking an educational approach to management of chronic tinnitus. The educational aspect is emphasized; thus, we named our approach tinnitus school. Selection of appropriate patients and follow-up is based on psychometrics of tinnitus and stress questionnaires, including a tinnitus reaction questionnaire, a tinnitus cognitive questionnaire, and a 20-item perceived stress questionnaire. Tinnitus school is a three-phase program: counseling, training, and home training. Training is based on a tinnitus-fitted physiotherapeutic protocol.


Impaired humoral immunity and tolerance in K14-VEGFR-3-Ig mice that lack dermal lymphatic drainage.
Lymphatic vessels transport interstitial fluid, soluble Ag, and immune cells from peripheral tissues to lymph nodes (LNs), yet the contribution of peripheral lymphatic drainage to adaptive immunity remains poorly understood. We examined immune responses to dermal vaccination and contact hypersensitivity (CHS) challenge in K14-VEGFR-3-Ig mice, which lack dermal lymphatic capillaries and experience markedly depressed transport of solutes and dendritic cells from the skin to draining LNs. In response to dermal immunization, K14-VEGFR-3-Ig mice produced lower Ab titers. In contrast, although delayed, T cell responses were robust after 21 d, including high levels of Ag-specific CD8+ T cells and production of IFN-γ, IL-4, and IL-10 upon restimulation. T cell-mediated CHS responses were strong in K14-VEGFR-3-Ig mice, but importantly, their ability to induce CHS tolerance in the skin was impaired. In addition, 1-y-old mice displayed multiple signs of autoimmunity. These data suggest that lymphatic drainage plays more important roles in regulating humoral immunity and peripheral tolerance than in effector T cell immunity.


One-hour plasma glucose identifies insulin resistance and beta-cell dysfunction in individuals with normal glucose tolerance: cross-sectional data from the Relationship between Insulin Sensitivity and Cardiovascular Risk (RISC) study.
Some individuals with normal glucose tolerance (NGT) exhibit a 1-h excursion of plasma glucose during oral glucose tolerance testing as high as that of individuals with impaired glucose tolerance (IGT). The aim of this study was to characterize their metabolic phenotype. A total of 1,205 healthy volunteers (aged 29-61 years) underwent assessment of 1) oral glucose tolerance and 2) insulin sensitivity (standardized euglycemic-hyperinsulinemic clamp), as part of the Relationship between Insulin Sensitivity and Cardiovascular Risk (RISC) study. One-hour plasma glucose correlated better than 2-h plasma glucose with total insulin secretion (r = 0.43), beta-cell glucose sensitivity (r = -0.46), and beta-cell rate sensitivity (r = -0.18). Receiver operating characteristic analysis identified 8.95 mmol/l as the best cutoff value for prediction of IGT from 1-h plasma glucose (sensitivity 77% and specificity 80%). Participants with NGT with 1-h plasma glucose >8.95 mmol/l had larger waist circumference, higher BMI, lower insulin sensitivity, higher fasting glucose, and higher insulin secretion than their counterparts with 1-h plasma glucose <or=8.95 mmol/l (P < 0.001 for all comparisons). Moreover, they exhibited lower beta-cell glucose sensitivity (P < 0.001), beta-cell rate sensitivity (P < 0.001), and potentiation factor (P = 0.026). When compared with conventionally defined IGT, they were not different in waist circumference and BMI, hepatic insulin extraction, beta-cell glucose sensitivity, beta-cell rate sensitivity, and potentiation factor but did have greater insulin sensitivity along with reduced basal (P = 0.001) and total insulin secretion (P = 0.002). Higher values of 1-h plasma glucose may identify an intermediate condition between NGT and IGT characterized by greater insulin resistance, reduced beta-cell glucose sensitivity, and reduced beta-cell rate sensitivity.


Differential effect of FHM2 mutation on synaptic plasticity in distinct hippocampal regions.
Familial hemiplegic migraine 2 is a pathology linked to mutation of the ATP1A2 gene producing loss of function of the α2 Na+/K+-ATPase (NKA). W887R/+ knock-in (KI) mice are used to model the familial hemiplegic migraine 2 condition and are characterized by 50% reduced NKA expression in the brain and reduced rate of K+ and glutamate clearance by astrocytes. These alterations might, in turn, produce synaptic changes in synaptic transmission and plasticity. Memory and learning deficits observed in familial hemiplegic migraine patients could be ascribed to a possible alteration of hippocampal neuronal plasticity and measuring possible changes of long-term potentiation in familial hemiplegic migraine 2 KI mice might provide insights to strengthen this link. Here we have investigated synaptic plasticity in distinct hippocampal regions in familial hemiplegic migraine 2 KI mice. We show that the dentate gyrus long-term potentiation of familial hemiplegic migraine 2 mice is abnormally increased in comparison with control animals. Conversely, in the CA1 area, KI and WT mice express long-term potentiation of similar amplitude. The familial hemiplegic migraine 2 KI mice show region-dependent hippocampal plasticity abnormality, which might underlie some of the memory deficits observed in familial migraine.


Phenotypic and genotypic characterization of lactic acid bacteria from traditional cheese in Khorramabad city of Iran with probiotic potential.
Lactic acid bacteria (LAB) with proteolitic activity are used as aromatic and antibacterial substances, cholesterol reduces, bile salt hydrolyses, and probiotic. The aims of this project were to isolate and identify natural LAB flora involved in traditional fermentation in cheeses of Khoramabad city and also to survey their probiotic potential. In order to achieve this goal, LAB were isolated and characterized using phenotypic and genotypic methods (PCR-sequencing); in the next stage, they were analyzed lowering cholesterol medium, hydrolysis of the bile, resistance to bile-resistant PH acidic stomach. At the end of the study, 88 cocci and 3 bacill were found: 58 Enterococcus faecium, 16 Enterococcus hirae, 5 Lactococcus lactis, 3 Lactobacillus plantarum, and 9 undetermined. The probiotic results of the bacteria had effects on the reduction of cholesterol, resistance to stomach acid, had relative antibacterial effects, and some strains had effects on hydrolyzing the bile. For further identification, the PCR method and the application of 16s-DNA-ITS genes and its sequencing were found useful. This study showed that lactic acid bacteria in the traditional cheese of the Khorramabad city have relative probiotic effect and that these lactic acid bacteria in fermented milk are suitable.


Posturally induced microvascular constriction in patients with different stages of leg ischaemia: effect of local skin heating.
1. Skin microcirculation was investigated in 12 asymptomatic subjects and 76 patients, grouped according to their ankle-to-brachial systolic blood pressure index, in order to evaluate to what extent posturally induced microvascular constriction is dependent on the stage of leg ischaemia at different local skin temperatures. 2. Skin microcirculation was assessed in the supine and sitting position by using laser Doppler fluxmetry at unheated skin temperature and at 36 degrees C, and transcutaneous oximetry at 37 degrees C and 44 degrees C. 3. Skin perfusion and oxygenation diminished with decreasing ankle-to-brachial systolic blood pressure index. In healthy control subjects, perfusion and oxygenation were reduced when changing from the supine to the sitting position, but were enhanced in patients with severe leg ischaemia (ankle-to-brachial systolic blood pressure less than 30%), indicating disturbed posturally induced vasoconstriction. 4. Increasing the local skin temperature resulted in a higher perfusion and masked the posturally induced vasoconstriction in healthy subjects. In patients with severe leg ischaemia, however, perfusion was unaltered by the temperature increase, apparently because the microvessels were already maximally dilated. The induction of reactive hyperaemia produced no additional increase in perfusion or oxygenation. 5. It is concluded that posturally induced microvascular constriction in the skin is disturbed in patients with severe leg ischaemia (ankle-to-brachial systolic blood pressure index less than 30%). Disturbed microvascular constriction upon dependency was also seen in healthy subjects after local skin heating. This suggests that posturally induced vasoconstriction is mainly regulated by local mechanisms.


Nuclear architecture of resting and LPS-stimulated porcine neutrophils by 3D FISH.
Neutrophils are essential components of the innate immune system due to their ability to kill and phagocytose invading microbes. They possess a lobulated nucleus and are capable of extensive and complex changes in response to bacterial stimulation. The aim of our study was to investigate whether the 3D nuclear organization of porcine neutrophils was modified upon stimulation. The organization of centromeres, telomeres, and chromosome territories (chromosomes 2, 3, 7, 8, 12, 13, and 17) was studied on structurally preserved nuclei using 3D fluorescence in situ hybridization, confocal microscopy, and image analysis. By differential labeling of centromeres of acrocentric and metacentric/submetacentric chromosomes, we showed that centromeres associated to form chromocenters but did so preferentially between chromosomes with the same morphology. Upon activation, some of these chromocenters dispersed. Telomeres were also found to form clusters, but their number remained unchanged in lipopolysaccharide-stimulated neutrophils. The analysis of the position of chromocenters and telomere clusters showed a more internal location of the latter compared to the former. The analysis of chromosome territories revealed that homologs were distributed randomly among lobes whatever the cell's status. The volume of these territories was not proportional to chromosome length, and some chromosomes (chr 3, 12, 13, and 17) were more prone to decondensation when neutrophils were stimulated. Thus, our study demonstrated that activation of neutrophils resulted in several modifications of their nuclear architecture: a decrease in the number of non-acrocentric chromocenters and the decondensation of several chromosomes.


Nonfatal systemic air embolism complicating percutaneous CT-guided transthoracic needle biopsy: four cases from a single institution.
Systemic air embolism is recognized as a potentially fatal but extremely rare complication following percutaneous transthoracic needle biopsy. However, its incidence might be underestimated by missing systemic air in patients without cardiac or cerebral symptoms. This study was based on four cases (one man and three women; age range, 54 to 75 years) of systemic air embolism complicating CT scan-guided transthoracic needle biopsy, which were encountered among 1,010 procedures performed at our institution from April 1999 to December 2006. The target lesion was a lung tumor in three patients, and a mediastinal tumor in one patient. The procedure was performed percutaneously under CT scan-fluoroscopic guidance by using a coaxial biopsy needle system. In all four patients, a specimen was successfully obtained from the lesions. During or immediately after the procedure, all patients experienced paroxysms of coughing. In three patients without cardiac or cerebral symptoms, the presence of systemic air was confirmed on postprocedural CT scan images; it was resolved without causing morbidity after the immediate therapy. The presence of systemic air was missed in one initially asymptomatic patient, resulting in a subsequent neurologic deficit. Systemic air embolism following CT scan-guided transthoracic needle biopsy was encountered more frequently than would be expected. The considerable attention we gave to this complication enabled us to recognize it in patients without cardiac or cerebral symptoms. No sequelae were observed in the three patients in whom systemic air embolism was detected, and the therapy was initiated immediately, whereas missing systemic air led to cerebral embolism in one patient in our four cases.


Low-coverage vaccination strategies for the conservation of endangered species.
The conventional objective of vaccination programmes is to eliminate infection by reducing the reproduction number of an infectious agent to less than one, which generally requires vaccination of the majority of individuals. In populations of endangered wildlife, the intervention required to deliver such coverage can be undesirable and impractical; however, endangered populations are increasingly threatened by outbreaks of infectious disease for which effective vaccines exist. As an alternative, wildlife epidemiologists could adopt a vaccination strategy that protects a population from the consequences of only the largest outbreaks of disease. Here we provide a successful example of this strategy in the Ethiopian wolf, the world's rarest canid, which persists in small subpopulations threatened by repeated outbreaks of rabies introduced by domestic dogs. On the basis of data from past outbreaks, we propose an approach that controls the spread of disease through habitat corridors between subpopulations and that requires only low vaccination coverage. This approach reduces the extent of rabies outbreaks and should significantly enhance the long-term persistence of the population. Our study shows that vaccination used to enhance metapopulation persistence through elimination of the largest outbreaks of disease requires lower coverage than the conventional objective of reducing the reproduction number of an infectious agent to less than one.


Phase II evaluations of cilengitide in asymptomatic patients with androgen-independent prostate cancer: scientific rationale and study design.
Two randomized trials demonstrated an improvement in survival with docetaxel-based chemotherapy for patients with metastatic, androgen-independent prostate disease. However, the effect of current therapy is suboptimal in that it is complicated by toxicities and has no curative potential. Cilengitide (EMD121974; NSC 707544), is a potent selective alphavbeta3 and alphavbeta5 integrin antagonist. Integrins are cell surface receptors that mediate a variety of cell activities including endothelial cell proliferation and migration. Blocking the ligation of integrins by antagonists promotes apoptosis of proliferative angiogenic cells, thereby suspending new blood vessel formation, which is essential for the growth of malignant disease. In prostate cancer specifically, integrins are known to be involved in metastases with differential expression on tumor cells. Tumors and vascular endothelial cells produce factors, such as vascular endothelial growth factor and basic fibroblast growth factor, that promote neovascularization, which has been implicated in prostate cancer progression. Cilengitide has been shown to inhibit alphavbeta3- and alphavbeta5-mediated cell adhesion and block in vitro endothelial cell migration. In vivo experiments demonstrated that cilengitide inhibited cytokine-induced basic fibroblast growth factor- and vascular endothelial growth factor-mediated angiogenesis in a dose-dependent manner. Cilengitide also inhibited tumor growth in various in vivo systems. Two Cancer Therapy Evaluation Program-sponsored, multicenter, phase II trials are designed to evaluate the safety and efficacy of this agent in patients with androgen-independent prostate cancer. National Cancer Institute trial 6735 is evaluating cilengitide at 2000 mg in patients with nonmetastatic androgen-independent prostate cancer, and National Cancer Institute trial 6372 is evaluating 2 dose levels of cilengitide, 500 mg or 2000 mg, intravenously twice weekly in patients with metastatic prostate cancer.


Photodynamic therapy inactivates extracellular matrix-basic fibroblast growth factor: insights to its effect on the vascular wall.
Photodynamic therapy (PDT), the light activation of photosensitizer dyes for the production of oxygen and other free radical moieties without the generation of heat, has been shown to inhibit the development of experimentally induced intimal hyperplasia. The host response to PDT, a form of vascular injury that results in complete vascular wall cell eradication, is devoid of inflammation and proliferation and promotes favorable vascular wall healing. These effects do not result in intimal hyperplasia and are suggestive of PDT-induced changes in the extracellular matrix (ECM). As a model to better understand the biologic consequences of PDT on the vascular wall matrix proteins, the effect of PDT was studied on the powerful matrix-resident mitogen basic fibroblast factor (bFGF) in vitro. PDT (5 to 200 J/cm2, 100 mW/cm2, 675 nm) was used with the photosensitizer chloroaluminum sulfonated phthalocyanine (5 micrograms/ml) to inactivate bFGF in vitro while 100 J/cm2 of irradiation was administered 24 hours after 5 mg/ml of the photosensitizer was used in vivo. PDT was used on bFGF in solution and on endothelial cell-derived ECM. Enzyme-linked immunosorbent assay was used to quantitate bFGF in solution after PDT treatment or after extraction from the ECM by collagenase and heparin. Functional activity of matrix-associated bFGF was assessed by smooth muscle cell mitogenesis by 3H-thymidine incorporation. To demonstrate the in vivo relevance of these observations, immunohistochemical analysis of PDT-treated rat carotid arteries was undertaken. PDT eliminated detectable levels of bFGF in solution. PDT of ECM significantly reduced matrix-bound bFGF (1.0 +/- 0.6 vs 27.5 +/- 1.3 pg/ml; p < 0.0001). This reduction in bFGF after PDT of the ECM was associated with a decrease in vascular smooth muscle cell mitogenesis (52.4% +/- 4.6%; p < 0.0001) when plated on PDT-treated matrix compared with nontreated matrix. Quantitative replenishment of exogenous bFGF to PDT-treated matrix restored proliferation to baseline levels. PDT of rat carotid arteries demonstrated a loss of bFGF staining compared with control nontreated arteries. PDT inactivation of matrix-resident bFGF and possibly other bioactive molecules can provide a mechanism by which PDT suppresses smooth muscle cell proliferation in the vessel wall. This free radical-mediated alteration of matrix may contribute to favorable vascular healing when PDT is used for the inhibition of injury-induced intimal hyperplasia.


A Review of and Algorithmic Approach to Soft Palate Reconstruction.
The soft palate contributes to deglutition, articulation, and respiration. Current reconstructive techniques focus on restoration of both form and function. The unique challenges of soft palate reconstruction include maintenance of complex upper aerodigestive tract function, with minimal local or donor site morbidity. To review the literature on soft palate reconstruction and present an algorithm on how to approach soft palate defects based on this review. A review of the literature for articles reporting studies on and that described concepts related to soft palate reconstruction was conducted in March 2017. In all, 1804 candidate titles and abstracts were independently reviewed. English-language articles that discussed acquired soft palate defect reconstruction were included. Non-English language studies without available translations, studies on primary soft palate defect reconstruction (ie, cleft palate repair) and primary cleft palate repair, studies in which the soft palate was not the focus of the article, and studies involving animals were excluded. The following observations were made from the review of 92 included articles. Soft palate anatomy is a complex interplay of multiple structures working in a 3-dimensional area. Three of the authors created an initial algorithmic framework based on the selected studies. After this, a round table discussion among 3 authors considered experts was used to refine the algorithm based on their expert opinion. The 4 most important factors were determined to be defect size, defect extension to other subsites, defect thickness, and history of radiotherapy or planned radiotherapy. This algorithm includes both surgical and nonsurgical options. Defects in the soft palate not only affect the size and shape of the organ but, more critically, the function. The reconstructive ladder is used to help maximize the remaining soft palate functional tissue and minimize the effect of nonfunctional implanted tissue. Partial-thickness defects or defects less than one-fourth of the soft palate may not require locoregional tissue transfer. Patients with a history of radiotherapy or defects of up to 75% of the soft palate may require locoregional tissue transfer. Defects greater than 75% of the soft palate, defects that include exposure of the neck vasculature, or defects that include significant portions of the hard palate or adjacent oropharyngeal subsites may require free tissue transfer. Obturation should be considered a second-line option in most cases. Ideal reconstruction of the soft palate relies on a comprehensive understanding of soft palate anatomy, a full consideration of the armamentarium of surgical techniques, consideration for adjacent subsite deficits, and a detailed knowledge of various intrinsic and extrinsic patient factors to optimize speech, swallowing, and airway outcomes. The included algorithm may serve as a useful starting point for the surgeon when considering reconstruction.


Surgical treatment of temporal lobe epilepsy: clinical, radiological, and histopathological findings in 178 patients.
The surgical treatment of pharmacoresistant temporal lobe epilepsy is increasing rapidly. The correlation of preoperative MRI, histopathological findings, and postoperative seizure control is reported for 178 patients with chronic medically intractable temporal lobe epilepsy who were operated on between November 1987 and January 1993. Histopathologically there were distinct structural abnormalities in 97.2% of the surgical specimens. Signal abnormalities on MRI were present in 98.7% of patients with neoplastic lesions (n = 79), 76.6% of patients with non-neoplastic focal lesions (n = 55), and 69.2% of patients with Ammon's horn sclerosis (n = 39). Overall, structural abnormalities were detected by MRI in 82.7% of all patients. The mean postoperative follow up period was three years. Some 92% of the patients benefited from surgery: 103 patients (61.7%) were seizure free, 26 (15.5%) had no more than two seizures a year, and 24 (14.4%) showed a reduction of seizure frequency of at least 75%. Fourteen patients (8.4%) had a < 75% reduction of seizure frequency. The percentage of patients who were completely free of seizures after operation was 68.5% for patients with neoplastic lesions, 66.7% for Ammon's horn sclerosis, and 54.0% for patients with non-neoplastic focal lesions. By contrast, none of the patients in whom histopathological findings were normal became seizure free postoperatively. The data show that the presence of focal lesions or Ammon's horn sclerosis as determined by histopathological examination is associated with improved postoperative seizure control compared with patients without specific pathological findings. Brain MRI was very sensitive in detecting neoplasms; however, its sensitivity and specificity were limited with respect to non-neoplastic focal lesions and Ammon's horn sclerosis. Improvement of imaging techniques may provide a more precise definition of structural lesions in these cases and facilitate limited surgical resections of the epileptogenic area rather than standardised anatomical resections.


Biological effect of tissue plasminogen activator (t-PA) and DNase intrapleural delivery in pleural infection patients.
Pleural infection (PI) is a major global disease with an increasing incidence, and pleural fluid (PF) drainage is essential for the successful treatment. The MIST2 study demonstrated that intrapleural administration of tissue plasminogen activator (t-PA) and DNase, or t-PA alone increased the volume of drained PF. Mouse model studies have suggested that the volume increase is due to the interaction of the pleura with the t-PA via the monocyte chemoattractant protein 1 (MCP-1) pathway. We designed a study to determine the time frame of drained PF volume induction on intrapleural delivery of t-PA±DNase in humans, and to test the hypothesis that the induction is mediated by the MCP-1 pathway. Data and samples from the MIST2 study were used (210 PI patients randomised to receive for 3 days either: t-PA and DNase, t-PA and placebo, DNase and placebo or double placebo). PF MCP-1 levels were measured by ELISA. One-way and two-way analysis of variance (ANOVA) with Tukey's post hoc tests were used to estimate statistical significance. Pearson's correlation coefficient was used to assess linear correlation. Intrapleural administration of t-PA±DNase stimulated a statistically significant rise in the volume of drained PF during the treatment period (days 1-3). No significant difference was detected between any groups during the post-treatment period (days 5-7). Intrapleural administration of t-PA increased MCP-1 PF levels during treatment; however, no statistically significant difference was detected between patients who received t-PA and those who did not. PF MCP-1 expression was not correlated to the drug given nor the volume of drained PF. We conclude that the PF volume drainage increment seen with the administration of t-PA does not appear to act solely via activation of the MCP-1 pathway.


Asimadoline in the treatment of irritable bowel syndrome.
Irritable bowel syndrome (IBS) represents one of the most common gastrointestinal disorders, with the diarrhea-predominant form (D-IBS) representing an area of high unmet medical need. One difficulty in identifying suitable treatments for IBS is that although there are animal models for the components of IBS, it is not clear whether animals are afflicted by the disease. In a recently completed Phase II study, the kappa-opioid agonist, asimadoline, was shown to be efficacious in a prospectively defined subgroup of D-IBS patients. This study confirmed a good safety profile for asimadoline. The chemistry, pharmacology, pharmacokinetics, pharmacodynamics and clinical safety and efficacy of asimadoline in relationship to IBS is reviewed. Papers were searched over the past 20 years using the PubMed database and key words 'asimadoline', 'kappa-opioid agonist' and 'irritable bowel syndrome'. Abstracts were reviewed and appropriate full papers were then evaluated. The reader will gain an appreciation of kappa-opioid agonists as IBS treatments. In a prospectively defined, clinically relevant patient subgroup, asimadoline shows efficacy in the treatment of D-IBS.


Annealing of gold nanostructures sputtered on polytetrafluoroethylene.
Gold nanolayers sputtered on polytetrafluoroethylene (PTFE) surface and their changes induced by post-deposition annealing at 100°C to 300°C are studied. Changes in surface morphology and roughness are examined by atomic force microscopy, electrical sheet resistance by two point technique, zeta potential by electrokinetic analysis and chemical composition by X-ray photoelectron spectroscopy (XPS) in dependence on the gold layer thickness. Transition from discontinuous to continuous gold coverage takes place at the layer thicknesses 10 to 15 nm and this threshold remains practically unchanged after the annealing at the temperatures below 200°C. The annealing at 300°C, however, leads to significant rearrangement of the gold layer and the transition threshold increases to 70 nm. Significant carbon contamination and the presence of oxidized structures on gold-coated samples are observed in XPS spectra. Gold coating leads to a decrease in the sample surface roughness. Annealing at 300°C of pristine PTFE and gold-coated PTFE results in significant increase of the sample surface roughness.


The distribution of ocular Chlamydia prevalence across Tanzanian communities where trachoma is declining.
Mathematical models predict an exponential distribution of infection prevalence across communities where a disease is disappearing. Trachoma control programs offer an opportunity to test this hypothesis, as the World Health Organization has targeted trachoma for elimination as a public health concern by the year 2020. Local programs may benefit if a single survey could reveal whether infection was headed towards elimination. Using data from a previously-published 2009 survey, we test the hypothesis that Chlamydia trachomatis prevalence across 75 Tanzanian communities where trachoma had been documented to be disappearing is exponentially distributed. We fit multiple continuous distributions to the Tanzanian data and found the exponential gave the best approximation. Model selection by Akaike Information Criteria (AICc) suggested the exponential distribution had the most parsimonious fit to the data. Those distributions which do not include the exponential as a special or limiting case had much lower likelihoods of fitting the observed data. 95% confidence intervals for shape parameter estimates of those distributions which do include the exponential as a special or limiting case were consistent with the exponential. Lastly, goodness-of-fit testing was unable to reject the hypothesis that the prevalence data came from an exponential distribution. Models correctly predict that infection prevalence across communities where a disease is disappearing is best described by an exponential distribution. In Tanzanian communities where local control efforts had reduced the clinical signs of trachoma by 80% over 10 years, an exponential distribution gave the best fit to prevalence data. An exponential distribution has a relatively heavy tail, thus occasional high-prevalence communities are to be expected even when infection is disappearing. A single cross-sectional survey may be able to reveal whether elimination efforts are on-track.


Impaired wound healing in mice deficient in a matricellular protein SPARC (osteonectin, BM-40).
SPARC is a matricellular protein involved in cell-matrix interactions. From expression patterns at the wound site and in vitro studies, SPARC has been implicated in the control of wound healing. Here we examined the function of SPARC in cutaneous wound healing using SPARC-null mice and dermal fibroblasts derived from them. In large (25 mm) wounds, SPARC-null mice showed a significant delay in healing as compared to wild-type mice (31 days versus 24 days). Granulation tissue formation and extracellular matrix protein production were delayed in small 6 mm SPARC-null wounds initially but were resolved by day 6. In in vitro wound-healing assays, while wild-type primary dermal fibroblasts showed essentially complete wound closure at 11 hours, wound closure of SPARC-null cells was incomplete even at 31 hours. Addition of purified SPARC restored the normal time course of wound closure. Treatment of SPARC-null cells with mitomycin C to analyze cell migration without cell proliferation showed that wound repair remained incomplete after 31 hours. Cell proliferation as measured by 3H-thymidine incorporation and collagen gel contraction by SPARC-null cells were not compromised. A significant delay in healing large excisional wounds and setback in granulation tissue formation and extracellular matrix protein production in small wounds establish that SPARC is required for granulation tissue formation during normal repair of skin wounds in mice. A defect in wound closure in vitro indicates that SPARC regulates cell migration. We conclude that SPARC plays a role in wound repair by promoting fibroblast migration and thus granulation tissue formation.


NIST System for Measuring the Directivity Index of Hearing Aids under Simulated Real-Ear Conditions.
The directivity index is a parameter that is commonly used to characterize the performance of directional hearing aids, and is determined from the measured directional response. Since this response is different for a hearing aid worn on a person as compared to when it is in a free field, directivity index measurements of hearing aids are usually done under simulated real-ear conditions. Details are provided regarding the NIST system for measuring the hearing aid directivity index under these conditions and how this system is used to implement a standardized procedure for performing such measurements. This procedure involves a sampling method that utilizes sound source locations distributed in a semi-aligned zone array on an imaginary spherical surface surrounding a standardized acoustical test manikin. The capabilities of the system were demonstrated over the frequency range of one-third-octave bands with center frequencies from 200 Hz to 8000 Hz through NIST participation in an interlaboratory comparison. This comparison was conducted between eight different laboratories of members of Working Group S3/WG48, Hearing Aids, established by Accredited Standards Committee S3, Bioacoustics, which is administered by the Acoustical Society of America and accredited by the American National Standards Institute. Directivity measurements were made for a total of six programmed memories in two different hearing aids and for the unaided manikin with the manikin right pinna accompanying the aids. Omnidirectional, cardioid, and bidirectional response patterns were measured. Results are presented comparing the NIST data with the reference values calculated from the data reported by all participating laboratories.


Wild bird surveillance around outbreaks of highly pathogenic avian influenza A(H5N8) virus in the Netherlands, 2014, within the context of global flyways.
Highly pathogenic avian influenza (HPAI) A(H5N8) viruses that emerged in poultry in east Asia since 2010 spread to Europe and North America by late 2014. Despite detections in migrating birds, the role of free-living wild birds in the global dispersal of H5N8 virus is unclear. Here, wild bird sampling activities in response to the H5N8 virus outbreaks in poultry in the Netherlands are summarised along with a review on ring recoveries. HPAI H5N8 virus was detected exclusively in two samples from ducks of the Eurasian wigeon species, among 4,018 birds sampled within a three months period from mid-November 2014. The H5N8 viruses isolated from wild birds in the Netherlands were genetically closely related to and had the same gene constellation as H5N8 viruses detected elsewhere in Europe, in Asia and in North America, suggesting a common origin. Ring recoveries of migratory duck species from which H5N8 viruses have been isolated overall provide evidence for indirect migratory connections between East Asia and Western Europe and between East Asia and North America. This study is useful for better understanding the role of wild birds in the global epidemiology of H5N8 viruses. The need for sampling large numbers of wild birds for the detection of H5N8 virus and H5N8-virus-specific antibodies in a variety of species globally is highlighted, with specific emphasis in north-eastern Europe, Russia and northern China.


Epidemiology and impact of a multifaceted approach in controlling central venous catheter associated blood stream infections outside the intensive care unit.
Outside ICUs, CVC-ABSIs epidemiology and the results of strategies for their prevention are not well known. The aim of this study was to investigate the epidemiology and the impact of a multifaceted "bundle" approach in controlling CVC-ABSIs outside ICU. From 1991 we performed prevalence studies of device and parenteral nutrition use, and prospective surveillance of all episodes of CVC-ABSIs in a 350-bed teaching hospital. CVC-ABSIs incidence/1,000 inpatient-days was calculated. An estimated CVC-ABSIs incidence/1,000 catheter-days was calculated based on the prevalence rates of catheter use and the total number of inpatient-days in each year. On november 2008, an education programme was instituted for care of catheter lines: reinforcing instructions in aseptic insertion technique, after care and hand-washing; in order to assess the adherence to these measures the quantity of alcohol-based hand-rub consumption/1,000 patient-days was quoted in litres. From January 2009, a checklist intervention for CVC insertion in ICU was started: hand hygiene, using full barrier precautions, cleaning the skin with alcoholic chlorhexidine, avoiding femoral access and removing unnecessary catheters. Compliance with the central line insertion checklist was measured by real-time audits and was achieved in 80% of cases. Prevalence of use of CVC and parenteral nutrition was similar throughout the study. We followed-up 309 CVC-ABSIs cases. Estimated CVC-ABSIs rate progressively increased to 15.1/1,000 catheter-days in 2008 (0.36/1,000 inpatient-days). After the intervention, the alcohol-based hand-rub consumption increased slightly and estimated CVC-ABSIs rate fell to 10.1 /1,000 catheter-days in last three years (0.19/1,000 inpatient-days), showing a 32.9% decrease. The infection rates achieved were lower in Internal Medicine wards: decreased from 14.1/1,000 catheter-days (0.17/patient-days) in 2008 to 5.2/1,000 catheter-days (0.05/1,000 inpatient-days) in last three years, showing a 63.1% decrease. In 2009, the estimated CVC-ABSIs incidence rate was significantly lower in the Internal Medicine ward compared to the Surgery ward: rate ratio (RR) = 0.14, 95%CI: 0.03-0.60), and within the Internal Medicine ward, the estimated CVC-ABSIs incidence rate was significantly lower in 2009 compared to 2008 (RR = 0.20, 95%CI: 0.04-0.91). The rate of CVC-ABSIs increased outside-ICU, and the implementation of multifaceted infection control programme decreased their clinical impact.


Impossible meals? The food and meal situation of flight attendants in Scandinavia - A qualitative interview study.
The working conditions of flight attendants (FAs) often involve extended and irregular working hours, short rest periods, difficulties in planning for breaks and high demands of service provision. Moreover, work schedules including early check-in, shifts during circadian low and time-zone transitions imply constant exposure to alterations in circadian systems and related health risks. The aim of this explorative study was to investigate how the organisation of work, time and place influence the food and meal situation of FAs when at work, focusing on patterns, form and social context of meals. The research questions posed were how food and meals at work were characterised and perceived among the FAs, and what strategies were adopted to manage the food and meal situation. Qualitative, semi-structured interviews were conducted with fourteen FAs working in Scandinavia. The results indicated that the organisation of work, time and place have a major influence on the meal situation at work, and how food and meals are perceived and managed by FAs. The work was defined as fragmented and inconsistent regarding time and place resulting in scattered meals and a more snack-based form of eating. The meal situation was characterised by irregularity as well as unpredictability. Eating took place when food was available and when there was enough time to eat, rather than being guided by hunger or social context. Various strategies such as eating in prevention, using emergency food, avoiding certain food and drinks or eating little or nothing at all were used to manage the unpredictability of the meal situation as well as the gap between organisational and individual times. The findings demonstrated the individual responsibility to solve the meal at work, e.g. to solve organisational times.


Tissue oxidative metabolism after extreme hemodilution with PEG-conjugated hemoglobin.
NADH-localized fluorometry was used as a noninvasive technique to monitor changes in the energy state of intact tissue (muscle and connective tissue), without anesthesia, as a function of blood plasma O(2)-carrying capacity in the hamster window chamber model. Acute moderate isovolemic hemodilution was induced by two isovolemic hemodilution steps: in the first step, 6% 70-kDa dextran (Dex70) was used to induce an acute anemic state (18% Hct); in the second step, exchange transfusion of polyethylene glycol (PEG) maleimide-conjugated Hb (4 g/dl, PEG-Hb) or Dex70 (6 g/dl) was used to reduce erythrocytes to 75% of baseline (11% Hct). PEG-Hb had six copies of PEG (5 kDa) conjugated to each human Hb (0.48 g PEG/g Hb) through extension arm-facilitated chemistry. Systemic parameters, microvascular perfusion, functional capillary density, intravascular and interstitial Po(2), and intracellular NADH fluorescence were monitored. Mean arterial blood pressure after extreme hemodilution was statistically significantly reduced for Dex70 compared with PEG-Hb. The presence of PEG-Hb in the circulation maintained positive acid-base balance. While microvascular blood flows were not different, functional capillary density was significantly higher for PEG-Hb than Dex70. Arteriolar Po(2) was higher in the presence of PEG-Hb than Dex70, but tissue and venular Po(2) were not different. Cellular energy metabolism (intracellular O(2)) in the tissues was improved with PEG-Hb. Moderate hemodilution to 18% Hct (6.4 g Hb/dl) brings tissue O(2) delivery to the verge of inadequacy. Extreme hemodilution to 11% Hct (3.7 g Hb/dl) produces tissue anoxia, and high-O(2)-affinity PEG-Hb (Po(2) at which blood is 50% saturated with O(2) = 4 Torr, 1.1 g Hb/dl) only partially decreases anaerobic metabolism without increasing tissue Po(2).


Histopathologic characteristics, recurrence patterns, and survival of 129 patients with desmoplastic melanoma.
Desmoplastic melanoma (DM) has been associated with higher local recurrence rates than other types of cutaneous melanoma. Current controversies regarding locoregional treatment strategies warrant further investigation. Retrospective review of a prospectively maintained melanoma database identified 129 patients with DM out of >12,500 melanoma patients referred for treatment from 1980 to 2003. Clinical and histopathologic characteristics, recurrence, and survival were analyzed. The median follow-up was 4.0 years. Of the 129 patients identified, 82 (63.6%) were male, and the median age was 55.2 years. American Joint Committee on Cancer staging was I, II, and III in 25.6%, 68.0%, and 6.4% of patients, respectively, and the mean tumor thickness was 4.42 mm. Overall survival was 76% at 5 years and 64% at 10 years; median survival was 13.0 years. A total of 51 patients (39.5%) experienced disease recurrence, with a median time to recurrence of 1.3 years. The first recurrence was local in 18 patients (14.0%), nodal in 18 patients (14.0%), and distant in 15 patients (11.6%), with median survivals of 6.7, 7.8, and 1.8 years, respectively. Statistically significant predictors of recurrence were a final positive margin status and stage, and predictors of overall survival were patient age and stage. Compared with other types of melanoma, DMs do demonstrate a tendency toward local recurrence, thus suggesting that narrower excision margins may not be appropriate in this population. Scrutiny of final surgical margins is critical to the local management of DM. In addition, the potential for regional nodal involvement must be considered at the time of diagnosis and during surveillance for disease recurrence.


The Current Status of Nursing Professionalism Among Nursing Faculty in Japan.
The faculty of nursing schools plays an important role in the successful execution of nursing education. Therefore, faculty behavior strongly affects the professional development of nurses. However, few studies have examined professional nursing behaviors from the perspective of nursing faculty. Members of nursing faculty in Japan were surveyed regarding their perspectives on behaviors related to professionalism. The model, Miller's Wheel of Professionalism in Nursing, was used as the theoretical framework. The Behavioral Inventory for Professionalism in Nursing (BIPN) was completed by 74 full-time nursing faculty who were currently working at 10 institutes of nursing education in Japan. The mean BIPN score for the participants was 11.56 (SD = 6.08) of a possible total of 27. The highest and lowest BIPN category scores were for "research development, use, and evaluation" and "community service," respectively. Professionalism was found to relate significantly to higher educational preparation (F = 32.17, p < .0001). The Tukey-Kramer multiple comparison test found a significant association between having a graduate degree and higher scores for professionalism (p < .0001). The Spearman correlation coefficient was significant and positive for the relationship between professionalism and both educational preparation (r = .85, p < .0001) and number of years as a nursing educator (r = .31, p = .0077). The results support the idea that a higher level of educational preparation and more years of experience as a member of a nursing faculty are associated with higher levels of nursing professionalism. The professional behavior scores suggest that "community service" is an issue that requires further improvement among Japanese nursing faculty. Awareness of extrinsic factors such as education is important to maximize nursing professionalism. The findings of this study may help nursing faculty continue their self-development.


Complete genomic RNA sequence of western equine encephalitis virus and expression of the structural genes.
The complete nucleotide sequence of the 71V-1658 strain of western equine encephalitis virus (WEE) was determined (minus 25 nucleotides from the 5' end). A 5' RACE reaction was used to sequence the 5' terminus from WEE strain CBA87. The deduced WEE genome was 11508 nucleotides in length, excluding the 5' cap nucleotide and 3' poly(A) tail. The nucleotide composition was 28% A, 25% C, 25% G and 22% U. Comparison with partial WEE sequences of strain 5614 (nsP2-nsP3 of the nonstructural region) and strain BFS1703 (26S structural region) revealed comparatively little variation; a total of 149 nucleotide differences in 8624 bases (1.7% divergence), of which only 28% (42 nucleotides) altered the encoded amino acids. Comparison of deduced nsP1 and nsP4 amino acid sequences from WEE with the corresponding proteins from eastern equine encephalitis virus (EEE) yielded identities of 84.9 and 83.8%, respectively. Previously uncharacterized stem-loop structures were identified in the nontranslated terminal regions. A cDNA clone of the 26S region encoding the structural polyprotein of WEE strain 71V-1658 was placed under the control of a cytomegalovirus promoter and transfected into tissue culture cells. The viral envelope proteins were functionally expressed in tissue culture, as determined by histochemical staining with monoclonal antibodies that recognize WEE antigens, thus, forming the initial step in the investigation of subunit vaccines to WEE.


Prospective grading of graft-versus-host disease after unrelated donor marrow transplantation: a grading algorithm versus blinded expert panel review.
In conjunction with a randomized trial of T-cell depletion versus conventional graft-versus-host disease (GVHD) prophylaxis, we assessed GVHD grading by comparing the transplant center 100-day score, a clinically calculated algorithm, and a blinded expert panel review (PR). Weekly skin, gut, and liver clinical staging; clinically verified differential diagnosis; biopsy information; cyclosporine levels; and initiation of treatment were reviewed and graded according to the consensus GVHD grading method modified by a prospectively determined grading algorithm that specified liver and gut downstaging if a differential diagnosis in that organ was identified. Transplant center (TC) determination of maximum grade was compared with the algorithm-calculated grade and the final expert PR. Of 404 patients reviewed, the TC grade concurred with the calculated algorithm grade in 72% (the algorithm upgraded 18% and downgraded 10%), whereas the TC grade agreed with the PR in 77% (the PR upgraded 12% and downgraded 11%). The calculated algorithm grade was nearly fully (92%) concordant with the final PR grade (the PR upgraded 0.7% and downgraded 7%). Blinded, duplicate reviews for quality control (n = 108) agreed with the initial review in 89% of cases. Algorithm and/or PR review reduced the TC-reported incidence of grade II (28% to 23%) and increased grade III (11% to 20%), whereas grade 0 (41% to 42%), grade I (13% to 12%), and grade IV (7% to 6%) were invariant. Recalculation of the algorithm grading without differential diagnosis downstaging reduced agreement with the TC to a small extent. The original algorithm changed 51 (13%) of 404 from grade 0 to II into grade III or IV or vice versa; calculation without the downgrade modified 44 cases (11%). Maximum acute GVHD grade had a major effect on 2-year disease-free survival, but assignment by TC, calculated algorithm, or final PR grade had little effect on survival within grades or grade categories 0 through II versus III or IV. We conclude that detailed and expert PR yields GVHD scoring that is internally consistent and reproducible with 89% concordance. Weekly recording of GVHD stage along with a calculated grading algorithm acknowledging differential diagnoses results in a final and maximum grade nearly fully concordant with the expert blinded PR. Multicenter prospective GVHD scoring using all available weekly staging and differential diagnosis data can be reliably assessed with a clinically relevant algorithm. This approach can thereby reduce investigator bias, facilitate comparison between centers, and perhaps eliminate the need for an expert PR. This technique should be used in future prospective studies of GVHD prophylaxis.


Association of asthma and bipolar disorder: a nationwide population-based study in Taiwan.
The relationship between asthma and bipolar disorder has received little research. We sought to investigate this in a large national sample. Previous studies have found mood changes after prednisone use in asthma patients, and we therefore also investigated this exposure in relation to bipolar disorder. Cases were identified from Taiwan׳s National Health Insurance Research Database with a new primary diagnosis of asthma (ICD-9:493) between 2000 and 2007. Case status required the presence of any inpatient diagnosis of asthma and/or at least one year diagnosis of asthma in outpatient service. These 46,558 cases were compared to 46,558 sex-, age-, residence- and insurance premium-matched controls and both groups were followed until the end of 2008 for first diagnosis of bipolar disorder (ICD-9 codes 296.0 to 296.16, 296.4 to 296.81 and 296.89). Competing risk adjusted Cox regression analyses were applied, adjusting for sex, age, residence, insurance premium, prednisone, hyperthyroidism, COPD (chronic obstructive pulmonary disease), Charlson comorbidity index, and hospital admission days for any disorder. Of the 93,116 subjects, 161 were ascertained as having bipolar disorder during a mean (SD) follow-up period of 5.7 (2.2) years. Asthma was an independent risk for bipolar disorder in the fully adjusted model. Higher daily dose of prednisone was a risk factor in asthma cases. The severity of asthma and bipolar disorder, and the route/duration of prednisone treatment were not evaluated. Asthma was associated with increased risk of bipolar disorder. Higher daily dose of prednisone was associated with a further increased risk.


Social network analysis predicts health behaviours and self-reported health in African villages.
The provision of healthcare in rural African communities is a highly complex and largely unsolved problem. Two main difficulties are the identification of individuals that are most likely affected by disease and the prediction of responses to health interventions. Social networks have been shown to capture health outcomes in a variety of contexts. Yet, it is an open question as to what extent social network analysis can identify and distinguish among households that are most likely to report poor health and those most likely to respond to positive behavioural influences. We use data from seven highly remote, post-conflict villages in Liberia and compare two prominent network measures: in-degree and betweenness. We define in-degree as the frequency in which members from one household are named by another household as a friends. Betweenness is defined as the proportion of shortest friendship paths between any two households in a network that traverses a particular household. We find that in-degree explains the number of ill family members, whereas betweenness explains engagement in preventative health. In-degree and betweenness independently explained self-reported health and behaviour, respectively. Further, we find that betweenness predicts susceptibility to, instead of influence over, good health behaviours. The results suggest that targeting households based on network measures rather than health status may be effective for promoting the uptake of health interventions in rural poor villages.


Altered muscle satellite cell activation following 16 wk of resistance training in young men.
Skeletal muscle satellite cells (SC) play an important role in muscle adaptation. In untrained individuals, SC content and activation status have been observed to increase in response to a single bout of exercise. Muscle fiber characteristics change considerably when resistance exercise is performed chronically, but whether training status affects the activity of SC in response to a single bout of exercise remains unknown. We examined the changes in SC content and activation status following a single bout of resistance exercise, before and following a 16-wk progressive resistance training (RT) program in 14 young (25 ± 3 yr) men. Before and after RT, percutaneous biopsies from the vastus lateralis muscle were taken before a single bout of resistance exercise and after 24 and 72 h of postexercise recovery. Muscle fiber size, capillarization, and SC response were determined by immunohistochemistry. Following RT, there was a greater activation of SC after 24 h in response to a single bout of resistance exercise (Pre, 1.4 ± 0.3; 24 h, 3.1 ± 0.3 Pax7+/MyoD+ cells per 100 fibers) compared with before RT (Pre, 1.4 ± 0.3; 24 h, 2.2 ± 0.3 Pax7+/MyoD+ cells per 100 fibers, P < 0.05); no difference was observed 72 h postexercise. Following 16 wk of RT, MyoD mRNA expression increased from basal to 24 h after the single bout of exercise (P < 0.05); this change was not observed before training. Individual capillary-to-fiber ratio (C/Fi) increased in both type I (1.8 ± 0.3 to 2.0 ± 0.3 C/Fi, P < 0.05) and type II (1.7 ± 0.3 to 2.2 ± 0.3 C/Fi, P < 0.05) fibers in response to RT. After RT, enhanced activation of SC in response to resistance exercise is accompanied by increases in muscle fiber capillarization.


Impact of ribavirin priming on viral kinetics and treatment response in chronic hepatitis C genotype 1 infection.
Ribavirin amplifies the interferon-alpha (IFN) signalling cascade. As ribavirin needs 4 weeks to reach steady state, ribavirin priming may optimize hepatic IFN sensitivity before starting a pegylated (PEG)-IFN/ribavirin combination therapy. This study investigated potential benefits of ribavirin priming prior to PEG-IFN2a/ribavirin combination therapy on viral kinetics, on-treatment and sustained virological response (SVR) in chronic hepatitis C virus (HCV) genotype 1 infection. Sixty-eight treatment naive patients were randomized 2:2:1 to ribavirin (ribavirin arm) or placebo (placebo arm) or PEG-IFN2a (PEG-IFN2a arm) for 6 weeks prior to 12 weeks of PEG-IFN2a/ribavirin combination therapy within a double-blind, placebo-controlled trial. Then, standard PEG-IFN2a/ribavirin combination therapy according to the German guidelines was continued under the responsibility of the investigators. Ribavirin was given according to body weight and PEG-IFN2a at a dose of 180 μg subcutaneously once/week. During ribavirin priming, HCV RNA showed a decline of -0.58 log10 IU/mL (P < 0.001) that was unrelated to the IL28B rs12979860 genotype (CC vs CT/TT, P = 0.244). Ribavirin priming did neither increase the PEG-IFN2a-induced first- or second-phase viral decline (P values >0.100) nor on-treatment response or SVR (HCV RNA undetectable at week 12 of combination therapy: ribavirin arm 56%, placebo arm 38%, PEG-IFN2a arm 50%; SVR: ribavirin arm 41%, placebo arm 54%, PEG-IFN2a arm 50%; P values >0.300). In conclusion, ribavirin monotherapy showed a significant antiviral activity that was not influenced by the IL28B genotype. Ribavirin priming prior to PEG-IFN2a/ribavirin combination therapy did neither increase the first- or second-phase viral decline nor on-treatment response or SVR.


INsPeCT: INtegrative Platform for Cancer Transcriptomics.
The emergence of transcriptomics, fuelled by high-throughput sequencing technologies, has changed the nature of cancer research and resulted in a massive accumulation of data. Computational analysis, integration, and data visualization are now major bottlenecks in cancer biology and translational research. Although many tools have been brought to bear on these problems, their use remains unnecessarily restricted to computational biologists, as many tools require scripting skills, data infrastructure, and powerful computational facilities. New user-friendly, integrative, and automated analytical approaches are required to make computational methods more generally useful to the research community. Here we present INsPeCT (INtegrative Platform for Cancer Transcriptomics), which allows users with basic computer skills to perform comprehensive in-silico analyses of microarray, ChIP-seq, and RNA-seq data. INsPeCT supports the selection of interesting genes for advanced functional analysis. Included in its automated workflows are (i) a novel analytical framework, RMaNI (regulatory module network inference), which supports the inference of cancer subtype-specific transcriptional module networks and the analysis of modules; and (ii) WGCNA (weighted gene co-expression network analysis), which infers modules of highly correlated genes across microarray samples, associated with sample traits, eg survival time. INsPeCT is available free of cost from Bioinformatics Resource Australia-EMBL and can be accessed at http://inspect.braembl.org.au.


Morphological and phylogenetic analyses of Pythium species in South Africa.
The genus Pythium is important in agriculture, since it contains many plant pathogenic species, as well as species that can promote plant growth and some that have biocontrol potential. In South Africa, very little is known about the diversity of Pythium species within agricultural soil, irrigation and hydroponic systems. Therefore, the aim of the study was to characterise a selection of 85 Pythium isolates collected in South Africa from 1991 through to 2007. The isolates were characterised morphologically as well as through sequence and phylogenetic analyses of the internal transcribed spacer regions (ITS) and the 5.8S gene of the nuclear ribosomal DNA. Phylogenetic analyses showed that the isolates represented ten of the 11 published Pythium clades [Lévesque & De Cock, 2004. Molecular phylogeny and taxonomy of the genus Pythium. Mycological Research 108: 1363-1383]. Characterisation of isolates in clade D and J suggested that the phylogenetic concept of Pythium acanthicum and Pythium perplexum respectively, needs further investigation in order to enable reliable species identification within these clades. Our phylogenetic analyses of Pythium species in clade B also showed that species with globose sporangia group basal within this clade, and are not dispersed within the clade as previously reported. The 85 South African isolates represented 34 known species, of which 20 species have not been reported previously in South Africa. Additionally, three isolates (PPRI 8428, 8300 and 8418) were identified that may each represent putative new species, Pythium sp. WJB-1 to WJB-3.


Quantitative Profiling of Protein S-Glutathionylation Reveals Redox-Dependent Regulation of Macrophage Function during Nanoparticle-Induced Oxidative Stress.
Engineered nanoparticles (ENPs) are increasingly utilized for commercial and medical applications; thus, understanding their potential adverse effects is an important societal issue. Herein, we investigated protein S-glutathionylation (SSG) as an underlying regulatory mechanism by which ENPs may alter macrophage innate immune functions, using a quantitative redox proteomics approach for site-specific measurement of SSG modifications. Three high-volume production ENPs (SiO2, Fe3O4, and CoO) were selected as representatives which induce low, moderate, and high propensity, respectively, to stimulate cellular reactive oxygen species (ROS) and disrupt macrophage function. The SSG modifications identified highlighted a broad set of redox sensitive proteins and specific Cys residues which correlated well with the overall level of cellular redox stress and impairment of macrophage phagocytic function (CoO > Fe3O4 ≫ SiO2). Moreover, our data revealed pathway-specific differences in susceptibility to SSG between ENPs which induce moderate versus high levels of ROS. Pathways regulating protein translation and protein stability indicative of ER stress responses and proteins involved in phagocytosis were among the most sensitive to SSG in response to ENPs that induce subcytoxic levels of redox stress. At higher levels of redox stress, the pattern of SSG modifications displayed reduced specificity and a broader set pathways involving classical stress responses and mitochondrial energetics (e.g., glycolysis) associated with apoptotic mechanisms. An important role for SSG in regulation of macrophage innate immune function was also confirmed by RNA silencing of glutaredoxin, a major enzyme which reverses SSG modifications. Our results provide unique insights into the protein signatures and pathways that serve as ROS sensors and may facilitate cellular adaption to ENPs, versus intracellular targets of ENP-induced oxidative stress that are linked to irreversible cell outcomes.


Estimating wearable motion sensor performance from personal biomechanical models and sensor data synthesis.
We present a fundamentally new approach to design and assess wearable motion systems based on biomechanical simulation and sensor data synthesis. We devise a methodology of personal biomechanical models and virtually attach sensor models to body parts, including sensor positions frequently considered for wearable devices. The simulation enables us to synthesise motion sensor data, which is subsequently considered as input for gait marker estimation algorithms. We evaluated our methodology in two case studies, including running athletes and hemiparetic patients. Our analysis shows that running speed affects gait marker estimation performance. Estimation error of stride duration varies between athletes across 834 simulated sensor positions and can soar up to 54%, i.e. 404 ms. In walking patients after stroke, we show that gait marker performance differs between affected and less-affected body sides and optimal sensor positions change over a period of movement therapy intervention. For both case studies, we observe that optimal gait marker estimation performance benefits from personally selected sensor positions and robust algorithms. Our methodology enables wearable designers and algorithm developers to rapidly analyse the design options and create personalised systems where needed, e.g. for patients with movement disorders.


Slow plasmon-polaritons in a bilayer metallic structure revealed by the lower-energy resonances of surface-enhanced Raman scattering.
Apart from the main plasmon-polariton resonance of the surface-enhanced Raman scattering (SERS) occurring at 480 - 530 nm, an additional resonance was observed for substrates with two silver layers separated by a dielectric layer which support extra plasmon modes with decreased group velocities. The novel SERS resonance is shifted towards lower energies and has comparable amplitude, its exact energy position being determined by the thickness of the dielectric interlayer. The experimental findings provide a ground for the engineering of SERS-substrates with the spectral position of the additional resonance matched with the photon energy of the pump laser over a fairly wide range of laser wavelengths.


Glyphosate and its formulation Roundup impair pig oocyte maturation.
Glyphosate, formulated as glyphosate-based herbicides (GBHs) including the best-known formulation Roundup, is the world's most widely used herbicide. During the last years, the growing and widespread use of GBHs has raised a great concern about the impact of environmental contamination on animal and human health including potential effect on reproductive systems. Using an in vitro model of pig oocyte maturation, we examined the biological impact of both glyphosate and Roundup on female gamete evaluating nuclear maturation, cytoplasmic maturation and developmental competence of oocytes, steroidogenic activity of cumulus cells as well as intracellular levels of glutathione (GSH) and ROS of oocytes. Our results indicate that although exposure to glyphosate and Roundup during in vitro maturation does not affect nuclear maturation and embryo cleavage, it does impair oocyte developmental competence in terms of blastocyst rate and cellularity. Moreover, Roundup at the same glyphosate-equivalent concentrations was shown to be more toxic than pure glyphosate, altering steroidogenesis and increasing oocyte ROS levels, thus confirming that Roundup adjuvants enhance glyphosate toxic effects and/or are biologically active in their side-effect and therefore should be considered and tested as active ingredients.