Weathering the storm: hurricanes and birth outcomes.
A growing literature suggests that stressful events in pregnancy can have negative effects on birth outcomes. Some of the estimates in this literature may be affected by small samples, omitted variables, endogenous mobility in response to disasters, and errors in the measurement of gestation, as well as by a mechanical correlation between longer gestation and the probability of having been exposed. We use millions of individual birth records to examine the effects of exposure to hurricanes during pregnancy, and the sensitivity of the estimates to these econometric problems. We find that exposure to a hurricane during pregnancy increases the probability of abnormal conditions of the newborn such as being on a ventilator more than 30min and meconium aspiration syndrome (MAS). Although we are able to reproduce previous estimates of effects on birth weight and gestation, our results suggest that measured effects of stressful events on these outcomes are sensitive to specification and it is preferable to use more sensitive indicators of newborn health.


Tissue-specific metabolic reprogramming drives nutrient flux in diabetic complications.
Diabetes is associated with altered cellular metabolism, but how altered metabolism contributes to the development of diabetic complications is unknown. We used the BKS db/db diabetic mouse model to investigate changes in carbohydrate and lipid metabolism in kidney cortex, peripheral nerve, and retina. A systems approach using transcriptomics, metabolomics, and metabolic flux analysis identified tissue-specific differences, with increased glucose and fatty acid metabolism in the kidney, a moderate increase in the retina, and a decrease in the nerve. In the kidney, increased metabolism was associated with enhanced protein acetylation and mitochondrial dysfunction. To confirm these findings in human disease, we analyzed diabetic kidney transcriptomic data and urinary metabolites from a cohort of Southwestern American Indians. The urinary findings were replicated in 2 independent patient cohorts, the Finnish Diabetic Nephropathy and the Family Investigation of Nephropathy and Diabetes studies. Increased concentrations of TCA cycle metabolites in urine, but not in plasma, predicted progression of diabetic kidney disease, and there was an enrichment of pathways involved in glycolysis and fatty acid and amino acid metabolism. Our findings highlight tissue-specific changes in metabolism in complication-prone tissues in diabetes and suggest that urinary TCA cycle intermediates are potential prognostic biomarkers of diabetic kidney disease progression.


Asymmetric synthesis of (+)-17-epi-methoxy-kauran-3-one through tandem oxidative polycyclization-pinacol process.
A synthesis of (+)-17-epi-methoxy-kauran-3-one, an O-methylated isomer of the natural diterpene 17-hydroxy-kauran-3-one, has been achieved. The strategy is based on a diastereoselective oxidative polycyclization-pinacol tandem process consisting of transforming a functionalized phenol into a compact and complex tetracycle, which represents the main core of kaurane family members. The synthesis also includes an enantioselective Yamamoto's allylation, a diastereoselective Ru-catalyzed hydrocyanation, a ring-closing metathesis and a reductive isomerization process as key steps. The structure of our synthetic substrate was determined through comparison with an O-methylated derivative of the natural compound.


The energy required to produce materials: constraints on energy-intensity improvements, parameters of demand.
In this paper, we review the energy requirements to make materials on a global scale by focusing on the five construction materials that dominate energy used in material production: steel, cement, paper, plastics and aluminium. We then estimate the possibility of reducing absolute material production energy by half, while doubling production from the present to 2050. The goal therefore is a 75 per cent reduction in energy intensity. Four technology-based strategies are investigated, regardless of cost: (i) widespread application of best available technology (BAT), (ii) BAT to cutting-edge technologies, (iii) aggressive recycling and finally, and (iv) significant improvements in recycling technologies. Taken together, these aggressive strategies could produce impressive gains, of the order of a 50-56 per cent reduction in energy intensity, but this is still short of our goal of a 75 per cent reduction. Ultimately, we face fundamental thermodynamic as well as practical constraints on our ability to improve the energy intensity of material production. A strategy to reduce demand by providing material services with less material (called 'material efficiency') is outlined as an approach to solving this dilemma.


An evaluation of an aftermarket local exhaust ventilation device for suppressing respirable dust and respirable crystalline silica dust from powered saws.
The objective of this study was to quantify the respirable dust and respirable silica exposures of roofing workers using an electric-powered circular saw with an aftermarket local exhaust ventilation attachment to cut concrete roofing tiles. The study was conducted to determine whether the local exhaust ventilation attachment was able to control respirable dust and respirable silica exposure below occupational exposure limits (OELs). Time-integrated filter samples and direct reading respirable dust concentrations were evaluated. The local exhaust ventilation consisted of a shroud attached to the cutting plane of the saw; the shroud was then connected to a small electric axial fan, which is intended to collect dust at the point of generation. All sampling was conducted with the control in use. Roofers are defined as those individuals who only lay tiles. Cutters/roofers are defined as those workers who operate the powered saw to cut tiles and also lay tiles. Respirable dust from this evaluation ranged from 0.13 to 6.59 milligrams per cubic meter (mg/m(3)) with a geometric mean of 0.38 mg/m(3) for roofers and from 0.45 to 3.82 mg/m(3) with a geometric mean of 1.84 mg/m(3) for cutters/roofers. Cutters/roofers usually handle areas close to crevices, edges, or tips of the roof whereas roofers handle areas where complete tiles can be placed. The respirable dust exposures for all cutters/roofers indicated concentrations exceeding the Occupational Safety and Health Administration's (OSHA) permissible exposure limit (PEL) for respirable dust containing silica; it was also exceeded for some of the roofers. The respirable silica concentrations ranged from 0.04 to 0.15 mg/m(3) with a geometric mean of 0.09 mg/m(3) for roofers, and from 0.13 to 1.21 mg/m(3) with a geometric mean of 0.48 mg/m(3) for cutters/roofers. As with respirable dust, the respirable silica exposures for cutters/roofers were higher than the exposures for roofers.


Signaling through the T1/ST2 molecule is not necessary for Th2 differentiation but is important for the regulation of type 1 responses in nonhealing Leishmania major infection.
T1/ST2 is a stable cell surface marker selectively expressed on type 2 T helper (Th2) effector cells. Since nonhealing Leishmania major infections in susceptible BALB/c mice have been ascribed to a polarized Th2 response, we used an anti-T1/ST2 monoclonal antibody (MAb) or a T1-Fc fusion protein to investigate the role of CD4+ T1/ST2(+) Th2 cells in experimental leishmaniasis. We show that interfering with T1/ST2 signaling had no effect on lesion development or parasite replication; however, it induced a significantly higher type 1 response and an enhanced capacity of CD4+ T cells to respond to interleukin 12 (IL-12). Surprisingly, even in the presence of an elevated Th1 response, the production of antigen-specific type 2 cytokines was not altered in the group of mice treated with the anti-T1/ST2 MAb or the T1-Fc fusion protein. To characterize further this Th2 response, we assessed the cytokine profile of CD4+ T cells and found that interfering with T1/ST2 signaling did not alter the cytokine profile of CD4+ T1/ST2(+) T cells. These results show that T1/ST2 signaling is not necessary for the differentiation of naive CD4+ T cells into antigen-specific CD4+ T1/ST2(+) Th2 cells. In addition to CD4+ T1/ST2(+) T cells, we detected another subpopulation of CD4+ Th2 cells, negative for the expression of T1/ST2, that could differentiate in vivo in response to L. major infection. Taken together, our results suggest that CD4+ T1/ST2(+) Th2 cells but not CD4+ T1/ST2(-) Th2 cells can downregulate the Th1 response during the course of a nonhealing L. major infection through a mechanism that is independent of IL-4 or IL-10.


Effect of different levels of briefly sustained ventricular pressure on arrhythmia in the isolated working rat heart.
1. The effect of different levels of ventricular pressure upon the prevalence of ventricular arrhythmias has been studied in 42 rat hearts by using the isolated working heart model. 2. The results have shown that there is an increased prevalence of arrhythmia at the highest levels of pressure tested in hearts from both normal and hypertensive rats. In hearts from normal rats total ectopic counts were 304 at 140 mmHg and 150 at 60 mmHg (P less than 0.05). In hearts from hypertensive rats total ectopic counts were 4217 at 180 mmHg and 2179 at 100 mmHg (P less than 0.05). 3. Hearts from hypertensive rats demonstrated significantly more ectopic activity than hearts from normal rats at all test pressures. Median values for ectopic counts for each study period were 68 in hearts from hypertensive rats and seven in hearts from normotensive rats (P less than 0.001). 4. There was evidence that the increased arrhythmia observed was not due to ischaemia or fatigue, and it seems likely that these effects of raised pressure are due to its effect on ventricular wall stress. The differences between various pressures were relatively small, but do suggest that a sustained increase in ventricular wall stress is arrhythmogenic. 5. This small arrhythmogenic effect over the relatively short period of these studies may be of much greater significance when considered in the context of the prolonged periods for which the diseased heart may be subjected to raised wall stress.


Perceived need for and use of mental health services by women living with or at risk of human immunodeficiency virus infection.
To determine prevalence of and factors associated with perceived need for and use of mental health services and use of psychotherapeutic medications among women in the Human Immunodeficiency Virus Epidemiology Research Study (HERS). We interviewed 871 HIV-seropositive and 439 demographically and behaviorally similar seronegative women at baseline regarding demographics, psychosocial measures, substance use, medical history, and use of health care services and medications. Thirty-eight percent of HIV-seropositive women and 35% of seronegative women (p = 0.33) reported needing mental health services in the prior six months. Women who were older, were white or Latina (compared with African American), had some college education, had less social support, and reported recent negative life events or had CES-D scores higher than 15 (suggesting depression) were more likely to report needing mental health services. Sixty-seven percent of seropositive and 65% of seronegative women who reported needing services (p = 0.69) reported obtaining services. Women who were white (compared with Latina or African American) and had more education, more social support, and health insurance were more likely to obtain services. Eighteen percent of seropositive and 13% of seronegative women (p = 0.006) reported current use of psychotherapeutic medications, especially antidepressants and antianxiety medications. Women who reported use of medications were more likely to report recent negative life events; were older, white, or Latina; had more education, less social support, CES-D scores higher than 15, and health insurance. Women both living with or at risk of HIV perceived a need for and used mental health services in association with negative life events, social isolation, and depressive symptoms. Need for and use of services differed by race/ethnicity, education, and availability of social support.


Concentrations of metals in water, sediment, biofilm, benthic macroinvertebrates, and fish in the Boulder River watershed, Montana, and the role of colloids in metal uptake.
To characterize the partitioning of metals in a stream ecosystem, concentrations of trace metals including As, Cd, Cu, Pb, and Zn were measured in water, colloids, sediment, biofilm (also referred to as aufwuchs), macroinvertebrates, and fish collected from the Boulder River watershed, Montana. Median concentrations of Cd, Cu, and Zn in water throughout the watershed exceeded the U.S. EPA acute and chronic criteria for protection of aquatic life. Concentrations of As, Cd, Cu, Pb, and Zn in sediment were sufficient in the tributaries to cause invertebrate toxicity. The concentrations of As, Cu, Cd, Pb, and Zn in invertebrates from lower Cataract Creek (63, 339, 59, 34, and 2,410 microg/g dry wt, respectively) were greater than the concentrations in invertebrates from the Clark Fork River watershed, Montana (19, 174, 2.3, 15, and 648 microg/g, respectively), that were associated with reduced survival, growth, and health of cutthroat trout fed diets composed of those invertebrates. Colloids and biofilm seem to play a critical role in the pathway of metals into the food chain and concentrations of As, Cu, Pb, and Zn in these two components are significantly correlated. We suggest that transfer of metals associated with Fe colloids to biological components of biofilm is an important pathway where metals associated with abiotic components are first available to biotic components. The significant correlations suggest that Cd, Cu, and Zn may move independently to biota (biofilm, invertebrates, or fish tissues) from water and sediment. The possibility exists that Cd, Cu, and Zn concentrations increase in fish tissues as a result of direct contact with water and sediment and indirect exposure through the food chain. However, uptake through the food chain to fish may be more important for As. Although As concentrations in colloids and biofilm were significantly correlated with As water concentrations, As concentrations in fish tissues were not correlated with water. The pathway for Pb into biological components seems to begin with sediment because concentrations of Pb in water were not significantly correlated with any other component and because concentrations of Pb in the water were often below detection limits.


Quantitative Multiple-Reaction Monitoring Proteomic Analysis of Gβ and Gγ Subunits in C57Bl6/J Brain Synaptosomes.
Gβγ dimers are one of the essential signaling units of activated G protein-coupled receptors (GPCRs). There are five Gβ and 12 Gγ subunits in humans; numerous studies have demonstrated that different Gβ and Gγ subunits selectively interact to form unique Gβγ dimers, which in turn may target specific receptors and effectors. Perturbation of Gβγ signaling can lead to impaired physiological responses. Moreover, previous targeted multiple-reaction monitoring (MRM) studies of Gβ and Gγ subunits have shown distinct regional and subcellular localization patterns in four brain regions. Nevertheless, no studies have quantified or compared their individual protein levels. In this study, we have developed a quantitative MRM method not only to quantify but also to compare the protein abundance of neuronal Gβ and Gγ subunits. In whole and fractionated crude synaptosomes, we were able to identify the most abundant neuronal Gβ and Gγ subunits and their subcellular localizations. For example, Gβ1 was mostly localized at the membrane while Gβ2 was evenly distributed throughout synaptosomal fractions. The protein expression levels and subcellular localizations of Gβ and Gγ subunits may affect the Gβγ dimerization and Gβγ-effector interactions. This study offers not only a new tool for quantifying and comparing Gβ and Gγ subunits but also new insights into the in vivo distribution of Gβ and Gγ subunits, and Gβγ dimer assembly in normal brain function.


Endogenous opioids in wound-site neutrophils of sternotomy patients.
Postoperative pain management is a critical aspect of patient care. The inflammatory state of the post-sternotomy surgical wound sensitizes nerve endings, causing pain. Unrelieved or improperly managed pain compromises wound healing. Peripheral opioid receptors play a major role in analgesia, particularly under inflammatory conditions where both opioid receptor expression and efficacy are increased. Leukocytic opioid peptides include β-endorphin (END), met-enkephalin (ENK), and dynorphin-A (DYN), with END and ENK being predominant. This work represents the first study of inflammatory cells collected from post-sternotomy wounds of patients undergoing cardiac surgery including coronary artery bypass grafting (CABG). Wound fluid (WF) and cells were collected from sternal wounds using a JP Blake drain at 24, 48, and 72 hours post sternum closure. Anti-CD15 staining and flow cytometry revealed that polymorphonuclear neutrophils (PMN) are the predominant cells present in wound fluid collected post-surgery. Compared to peripheral blood (PB) derived PMN, significant increases in CD177+/CD66b+ PMN were observed suggesting activation of wound-site PMN. Such activation was associated with higher levels of opioid peptide expression in PMN derived from WF. Indeed, increased level of opioid peptides in sternal wound environment was noted 72 h post-surgery. We demonstrate that WF contains factors that can significantly induce POMC transcription in human PMNs. IL-10 and IL-4 were abundant in WF and both cytokines significantly induced POMC gene expression suggesting that WF factors such as IL-10 and IL-4 contribute towards increased opioid peptide expression in wound-site PMN. This approach provided a unique opportunity to study the cross-talk between inflammation and opioid peptides in PMN at a sternotomy wound-site. Wound-site PMN exhibited induction of END and ENK. In addition, sternal wound fluid significantly induced END expression in PMN. Taken together, these data constitute first clinical evidence that human wound-site PMNs are direct contributors of opioids at the sternal wound-site.


A survey of the management, control, and complications of diabetes mellitus in patients attending a diabetes clinic in Blantyre, Malawi, an area of high HIV prevalence.
The aim of this study was to describe the current status of diabetes care in an urban diabetes clinic in Malawi and the prevalence of human immunodeficiency virus (HIV) in this population, investigating possible associations between HIV and diabetes. A systematic prospective survey of patients attending the diabetes clinic at a teaching hospital in Blantyre, Malawi was conducted. Six hundred twenty patients were assessed. Seventy-four percent had glycosylated hemoglobin (HbA1C) > 7.5%. Systolic blood pressure was > 140 mm Hg in 52% of patients. Hypertension was more common in patients with raised creatinine (P < 0.003), retinopathy (P = 0.01), and stroke (P < 0.0002). Microvascular complication rates were high, specifically nephropathy (34.7%), retinopathy (34.7%), and neuropathy (46.4%). HIV seroprevalence was 13.7%. HIV-positive subjects had a lower body mass index (BMI) and lower fasting blood sugar, and they were more likely to have albuminuria (48.0% versus 33.3%; P < 0.05). Control of glycemia and hypertension were poor, and microvascular complications were common. Nephropathy in diabetic patients may be affected by HIV status.


Brain gene expression in relation to fearfulness in female red junglefowl (Gallus gallus).
The biology of fear is central to animal welfare and has been a major target for selection during domestication. Fear responses were studied in female red junglefowl (RJF), the ancestor of domesticated chickens. A total of 31 females were tested in a ground predator test, an aerial predator test and a tonic immobility (TI) test, in order to assess their level of fearfulness across different situations. Two to six variables from each test were entered into a principal component (PC) analysis, which showed one major fearfulness component (explaining 27% of the variance). Based on the PC scores, four high- and four low-fearful birds were then selected for gene expression analysis. From each of these birds, the midbrain region (including thalamus, hypothalamus, pituitary, mesencephalon, pons, nucleus tractus solitarii and medulla oblongata), was collected and global gene expression compared between groups using a 14k chicken cDNA microarray. There were 13 significantly differentially expressed (DE) genes (based on M > 1 and B > 0; FDR-adjusted P < 0.05) between the fearful and non-fearful females. Among the DE genes, we identified the neuroprotein Axin1, two potential DNA/RNA regulating proteins and a retrotransposon transcript situated in a well-studied quantitative trait loci (QTL) region on chromosome 1, known to affect several domestication-related traits. The differentially expressed genes may be part of a possible molecular mechanism controlling fear responses in fowl.


Abdominal Angiostrongyliasis: A Presentation of Eosinophilic Granulomatous Colitis.
We present a case of a 4-year-old girl with abdominal angiostrongyliasis who presented with persistent fevers, hepatosplenomegaly, acute abdominal pain, and eosinophilia. Computed tomography scan identified thickening of the ascending colon with a narrowed lumen. Endoscopic evaluation revealed ulcerations and erythema in the ascending colon. The microscopic findings in biopsies included active chronic inflammation with prominent eosinophils and granulomas. A subset of granulomas contained the eggs of Angiostrongylus costaricensis. The definitive method of diagnosing A costaricensis is histology; peripheral blood serology has low specificity and the stool from infected patients does not contain eggs or larvae. Pathologists from endemic regions (Central and South America) are familiar with the typical histologic changes; however, because of increasing global travel, all pathologists should become familiar with A costaricensis, which may mimic common gastrointestinal diseases such as Crohn's disease, appendicitis, and Meckel's diverticulum.


Protective role of Nigella sativa in diabetic nephropathy: A randomized clinical trial.
This study aims to evaluate efficacy and safety of Nigella sativa oil supplementation in patients with chronic kidney disease Stage 3 and 4 due to diabetic nephropathy. It was a prospective, comparative, and open-label study. Patients were randomized into two groups. Group 1 (Control) received conservative management of diabetic nephropathy, whereas Group 2 (Test) received N. sativa oil (2.5 mL, once daily and per orally) along with conservative management for 12 weeks. Blood glucose, hemogram, and kidney function test were done at 0, 6, and 12 weeks of treatment. Significance of differences between pre- and post-treatment values in each group was assessed using Student's paired t-test and between the groups using unpaired t-test. We found a drop in blood glucose, serum creatinine, blood urea, and 24 h total urinary protein levels and a rise in glomerular filtration rate, 24 h total urinary volume, and hemoglobin level in the treatment group compared to the control group.


Flow characteristics of the proximal pulmonary arteries and vena cava in patients with chronic thromboembolic pulmonary hypertension: correlation between 3.0 T phase-contrast MRI and right heart catheterization.
We aimed to determine the correlation between flow characteristics of the proximal pulmonary arteries and vena cava obtained by 3.0 T phase-contrast magnetic resonance imaging (MRI) and hemodynamic characteristics by right heart catheterization in patients with chronic thromboembolic pulmonary hypertension. Twenty consecutive patients with chronic thromboembolic pulmonary hypertension and 20 sex- and age-matched healthy volunteers were included prospectively. All patients and controls underwent phase-contrast MRI to determine the flow characteristics including peak velocity, mean velocity, and mean blood flow of the proximal pulmonary artery and vena cava. All patients underwent right heart catheterization to determine the hemodynamics. Peak velocity and mean velocity of the proximal pulmonary artery were significantly lower in the patient group. In patients, both peak velocity and mean blood flow were sequentially decreased in the main pulmonary artery, left and right pulmonary arteries, and left and right interlobar pulmonary arteries. Inferior vena cava had higher peak velocity, mean velocity, and mean blood flow than superior vena cava. Peak velocity of the main pulmonary artery correlated with mean and diastolic pulmonary artery pressure. Peak velocity of both inferior and superior vena cava strongly correlated with the pulmonary vascular resistance index (PVRI) (r=-0.68, P < 0.001 and r=-0.74, P < 0.001, respectively). Mean velocity of the main pulmonary artery and right pulmonary artery strongly correlated with PVRI and mean pulmonary artery pressure. Mean velocity of the superior vena cava and mean blood flow of the main pulmonary artery strongly correlated with PVRI and right cardiac work index. Blood flow in the proximal pulmonary artery and vena cava evaluated by phase-contrast MRI correlate with hemodynamic parameters of right heart catheterization and can be used to noninvasively evaluate the severity of chronic thromboembolic pulmonary hypertension and, potentially, to follow up the treatment response.


Predictive models for mortality after ruptured aortic aneurysm repair do not predict futility and are not useful for clinical decision making.
The clinical decision-making utility of scoring algorithms for predicting mortality after ruptured abdominal aortic aneurysms (rAAAs) remains unknown. We sought to determine the clinical utility of the algorithms compared with our clinical decision making and outcomes for management of rAAA during a 10-year period. Patients admitted with a diagnosis rAAA at a large university hospital were identified from 2005 to 2014. The Glasgow Aneurysm Score, Hardman Index, Vancouver Score, Edinburgh Ruptured Aneurysm Score, University of Washington Ruptured Aneurysm Score, Vascular Study Group of New England rAAA Risk Score, and the Artificial Neural Network Score were analyzed for accuracy in predicting mortality. Among patients quantified into the highest-risk group (predicted mortality >80%-85%), we compared the predicted with the actual outcome to determine how well these scores predicted futility. The cohort comprised 64 patients. Of those, 24 (38%) underwent open repair, 36 (56%) underwent endovascular repair, and 4 (6%) received only comfort care. Overall mortality was 30% (open repair, 26%; endovascular repair, 24%; no repair, 100%). As assessed by the scoring systems, 5% to 35% of patients were categorized as high-mortality risk. Intersystem agreement was poor, with κ values ranging from 0.06 to 0.79. Actual mortality was lower than the predicted mortality (50%-70% vs 78%-100%) for all scoring systems, with each scoring system overestimating mortality by 10% to 50%. Mortality rates for patients not designated into the high-risk cohort were dramatically lower, ranging from 7% to 29%. Futility, defined as 100% mortality, was predicted in five of 63 patients with the Hardman Index and in two of 63 of the University of Washington score. Of these, surgery was not offered to one of five and one of two patients, respectively. If one of these two models were used to withhold operative intervention, the mortality of these patients would have been 100%. The actual mortality for these patients was 60% and 50%, respectively. Clinical algorithms for predicting mortality after rAAA were not useful for predicting futility. Most patients with rAAA were not classified in the highest-risk group by the clinical decision models. Among patients identified as highest risk, predicted mortality was overestimated compared with actual mortality. The data from this study support the limited value to surgeons of the currently published algorithms.


Microscopic lithography with pixelate diffraction of a digital micro-mirror device for micro-lens fabrication.
The irradiance in microscopic lithography using a digital micro-mirror device (DMD) as a virtual digital mask generator is influenced by diffraction effects that have been exploited to fabricate microstructures. Based on the established model, the theoretical analysis and simulation of DMD diffraction characteristics has been studied. A novel method without masking to fabricate a micro-lens by pixilation of micro-mirrors inside the DMDs used in microscopic lithography has been proposed. It is a method of precise control of photon-induced curing behavior of photoresist by full use of diffraction effects and verification of the feasibility of the fabrication method based on diffraction. The introduced method provides an option for accurate and flexible micro-fabrication of microstructures.


Extremes of cerebral blood flow during hypercapnic squat-stand maneuvers.
Squat-stand maneuvers (SSMs) are a popular method of inducing blood pressure (BP) oscillations to reliably assess dynamic cerebral autoregulation (dCA), but their effects on the cerebral circulation remain controversial. We designed a protocol whereby participants would perform SSMs under hypercapnic conditions. Alarmingly high values of cerebral blood flow velocity (CBFV) were recorded, leading to early study termination after the recruitment of a single participant. One healthy subject underwent recordings at rest (5 min sitting, 5 min standing) and during two SSMs (fixed and random frequency). Two sets of recordings were collected; one while breathing room air, one while breathing 5% CO2 . Continuous recordings of bilateral CBFV (transcranial Doppler), heart rate (ECG), BP (Finometer), and end-tidal CO2 (capnography) were collected. Peak values of systolic CBFV were significantly higher during hypercapnia (p < 0.01), and maximal values exceeded 200 cm.s-1 . Estimates of dCA (ARI) during hypercapnia were impaired relative to poikilocapnia (p = 0.03). The phase was significantly reduced under hypercapnic conditions (p = 0.03). Here we report extremely high values of CBFV in response to repeated SSMs during induced hypercapnia, in an otherwise healthy subject. Our findings suggest that protocols performing hypercapnic SSMs are potentially dangerous. We, therefore, urge caution if other research groups plan to undertake similar protocols.


Concentration-dependent differential effects of udenafil on viability, proliferation, and apoptosis in vascular endothelial and smooth muscle cells.
Local strategies directed against vascular smooth muscle cell (VSMC) proliferation, such as drug-eluting stents (DES), reduce the occurrence of restenosis. However, these approaches may also inhibit vascular endothelial cell (VEC) proliferation and impair reendothelialization, and hence, increase susceptibility to late thrombosis. In this study we examined the differential effects of various concentrations of the type 5 phosphodiesterase (PDE-5) inhibitor, udenafil, on viability, proliferation, and apoptosis of VEC and VSMC, in order to identify the optimal concentration of udenafil that minimizes inhibition of VEC survival and growth, and maximizes inhibition of VSMC survival and growth. VEC from human umbilical veins and VSMC from human aorta were exposed to various concentrations of udenafil (1, 10, and 100 μmol/l and 1 mmol/l) for 24 h, and its effects on cell viability, proliferation, and apoptosis were studied using 5-bromo-2'- deoxyuridine (BrdU), methylthiazoletetrazolium (MTT) assay, trypan blue dye exclusion, and flow cytometry. Udenafil inhibited the survival and growth of VEC and VSMC in a concentration-dependent manner over a range of concentrations. At 100 μmol/l, udenafil, inhibited VEC proliferation significantly less than VSMC proliferation (P < 0.05), and could significantly induce VEC apoptosis less than VSMC apoptosis (P < 0.05). Udenafil has a differential effect on survival and growth in VEC and VSMC. The maximal differential effect, with minimal inhibition of VEC and maximal inhibition of VSMC, occurs at 100 μmol/l. This characteristic suggests that udenafil is a promising agent for use in DES.


Influence of female sports dietitians' physical characteristics on athlete perception of effectiveness.
Research in sports-science disciplines such as sport psychology has demonstrated that practitioners' physical characteristics influence clients' perceptions of their effectiveness, potentially mediating the efficacy of subsequent interventions. However, very little research has been directed toward this issue for sports dietitians (SDs), the health professionals whom athletes are likely to engage to assist with manipulation of traits of physique. Therefore, the purpose of this investigation was to determine whether SDs' phenotype, specifically body-mass index (BMI), and type of dress influence potential clients' preference to consult them for dietetic support and if this affects their perceived effectiveness. One hundred volunteers (mean age 18.7 ± 0 .8 years) all participating in regular competitive sport, classified by gender (male, n = 55, or female, n = 45) and competitive standard (elite/subelite, n = 68, or club/recreational, n = 32) viewed slides representing four concurrently presented computer-generated images of the same female SD manipulated to represent different BMIs and dress types. Participants were asked to rank the SDs in order of their preference to work with them and, second, to rate their perceived effectiveness of each of the SDs. Key findings included the observation of a significant BMI main effect F(6, 91) = 387.39, p < .001 (effect size .96), with participants' ranking of preference and rating of perceived effectiveness of female SDs decreasing with increasing BMI. SDs should consider their physical appearance when meeting with athletes, as this may affect their perceived efficacy.


Intraseptal procaine abolishes hypothalamic stimulation-induced wheel-running and hippocampal theta field activity in rats.
Rats were implanted chronically with hippocampal recording electrodes, a microinfusion guide cannula aimed at the medial septal nucleus, and an electrode for electrical stimulation of the posterior hypothalamic nucleus (PH). PH stimulation elicited running in rats placed in a wheel and simultaneously occurring hippocampal theta field activity (HPC-theta). In the preprocaine (PRE) testing condition, a positive linear relationship was demonstrated among the intensity of electrical stimulation of the PH, wheel-running speed, and the peak frequency of HPC-theta. HPC-theta amplitude reached an asymptote at the lowest levels of electrical stimulation of the PH. Procaine hydrochloride (1.5 microliters, 20% solution), a local anesthetic, was then infused into the medial septal nucleus (MS). Five minutes after the infusion, PH stimulation no longer induced wheel-running behavior or HPC-theta, and the remaining irregular field activity was significantly reduced in amplitude. Fifteen minutes after the procaine infusion, PH stimulation still did not elicit HPC-theta or running behavior in the majority of animals but did evoke large-amplitude sharp-waves. Thirty minutes after the procaine infusion, PH stimulation again elicited HPC-theta and running behavior, but HPC-theta peak frequency and running speeds were both significantly reduced compared with PRE values. Forty-five minutes after the infusion, HPC-theta amplitude had recovered to PRE values, but HPC-theta frequency and running speeds elicited by PH stimulation were still significantly reduced. By 60 min after procaine administration, the amplitude and frequency of HPC-theta and the running speeds elicited by PH stimulation recovered to PRE values. Multiple regression analysis revealed that the recovery pattern of running behavior reflected the frequency rather than the amplitude of HPC-theta. Neither saline control infusions into the MS nor procaine infusions into the lateral septum and paraventricular thalamic nucleus affected HPC-theta or running behavior. These findings are consistent with the notion that both the locomotor activity and "movement-related" HPC-theta frequency induced by electrically stimulating the PH were attributable to ascending activation of a hypothalamo-septal pathway and not to activation of descending brainstem or peripheral motor systems.


The role of competition - colonization tradeoffs and spatial heterogeneity in promoting trematode coexistence.
Competition - colonization tradeoffs occur in many systems, and theory predicts that they can strongly promote species coexistence. However, there is little empirical evidence that observed competition- colonization tradeoffs are strong enough to maintain diversity in natural systems. This is due in part to a mismatch between theoretical assumptions and biological reality in some systems. We tested whether a competition - colonization tradeoff explains how a diverse trematode guild coexists in California horn snail populations, a system that meets the requisite criteria for the tradeoff to promote coexistence. A field experiment showed that subordinate trematode species tended to have higher colonization rates than dominant species. This tradeoff promoted coexistence in parameterized models but did not fully explain trematode diversity and abundance, suggesting a role of additional diversity maintenance mechanisms. Spatial heterogeneity is an alternative way to promote coexistence if it isolates competing species. We used scale transition theory to expand the competition - colonization tradeoff model to include spatial variation. The parameterized model showed that spatial variation in trematode prevalence did not isolate most species sufficiently to explain the overall high diversity, but could benefit some rare species. Together, the results suggest that several mechanisms combine to maintain diversity, even when a competition - colonization tradeoff occurs.


KinView: a visual comparative sequence analysis tool for integrated kinome research.
Multiple sequence alignments (MSAs) are a fundamental analysis tool used throughout biology to investigate relationships between protein sequence, structure, function, evolutionary history, and patterns of disease-associated variants. However, their widespread application in systems biology research is currently hindered by the lack of user-friendly tools to simultaneously visualize, manipulate and query the information conceptualized in large sequence alignments, and the challenges in integrating MSAs with multiple orthogonal data such as cancer variants and post-translational modifications, which are often stored in heterogeneous data sources and formats. Here, we present the Multiple Sequence Alignment Ontology (MSAOnt), which represents a profile or consensus alignment in an ontological format. Subsets of the alignment are easily selected through the SPARQL Protocol and RDF Query Language for downstream statistical analysis or visualization. We have also created the Kinome Viewer (KinView), an interactive integrative visualization that places eukaryotic protein kinase cancer variants in the context of natural sequence variation and experimentally determined post-translational modifications, which play central roles in the regulation of cellular signaling pathways. Using KinView, we identified differential phosphorylation patterns between tyrosine and serine/threonine kinases in the activation segment, a major kinase regulatory region that is often mutated in proliferative diseases. We discuss cancer variants that disrupt phosphorylation sites in the activation segment, and show how KinView can be used as a comparative tool to identify differences and similarities in natural variation, cancer variants and post-translational modifications between kinase groups, families and subfamilies. Based on KinView comparisons, we identify and experimentally characterize a regulatory tyrosine (Y177PLK4) in the PLK4 C-terminal activation segment region termed the P+1 loop. To further demonstrate the application of KinView in hypothesis generation and testing, we formulate and validate a hypothesis explaining a novel predicted loss-of-function variant (D523NPKCβ) in the regulatory spine of PKCβ, a recently identified tumor suppressor kinase. KinView provides a novel, extensible interface for performing comparative analyses between subsets of kinases and for integrating multiple types of residue specific annotations in user friendly formats.


Identification of the myotoxic site of the Lys49 phospholipase A(2) from Agkistrodon piscivorus piscivorus snake venom: synthetic C-terminal peptides from Lys49, but not from Asp49 myotoxins, exert membrane-damaging activities.
Group II phospholipase A(2) (PLA(2)) myotoxins found in the venoms of Crotalidae snakes can be divided into 'Asp49' and 'Lys49' isoforms, the latter being considered catalytically-inactive variants. Previous studies on one Lys49 isoform, myotoxin II from Bothrops asper, indicated that its myotoxic activity is due to the presence of a short cationic/hydrophobic sequence (115-129) near its C-terminus, which displays membrane-damaging properties. Since the C-terminal region of different group II PLA(2) myotoxins presents considerable sequence variability, synthetic peptides homologous to region 115-129 of myotoxin II, but corresponding to B. asper myotoxin III (Asp49), Agkistrodon piscivorus piscivorus Asp49 PLA(2) and Lys49 PLA(2), were studied to determine the possible functional relevance of such region for the toxic activities of these proteins. Results showed that both Lys49-derived peptides (p-BaK49 and p-AppK49) were able to lyse skeletal muscle C2C12 cells in culture, and to induce edema in the mouse footpad assay. Moreover, p-AppK49, which showed a markedly stronger cytotoxic potency than p-BaK49, additionally induced skeletal muscle necrosis when injected into mice. These observations unequivocally identify the sequence 115-129 (KKYKAYFKLKCKK) of the Lys49 PLA(2) of A. p. piscivorus as containing the key structural determinants needed for myotoxicity, and represent the first report of an unmodified, PLA(2)-derived short synthetic peptide with the ability to reproduce this effect of a parent toxin in vivo. On the other hand, the two Asp49-derived peptides did not show any toxic effects in vitro or in vivo, even at high concentrations. These findings suggests that Lys49 and Asp49 group II PLA(2)s might exert their myotoxic actions through different molecular mechanisms, by implying that the latter may not utilize their C-terminal regions as main membrane-destabilizing elements.


Spatio-temporal pattern of vestibular information processing after brief caloric stimulation.
Processing of vestibular information at the cortical and subcortical level is essential for head and body orientation in space and self-motion perception, but little is known about the neural dynamics of the brain regions of the vestibular system involved in this task. Neuroimaging studies using both galvanic and caloric stimulation have shown that several distinct cortical and subcortical structures can be activated during vestibular information processing. The insular cortex has been often targeted and presented as the central hub of the vestibular cortical system. Since very short pulses of cold water ear irrigation can generate a strong and prolonged vestibular response and a nystagmus, we explored the effects of this type of caloric stimulation for assessing the blood-oxygen-level-dependent (BOLD) dynamics of neural vestibular processing in a whole-brain event-related functional magnetic resonance imaging (fMRI) experiment. We evaluated the spatial layout and the temporal dynamics of the activated cortical and subcortical regions in time-locking with the instant of injection and were able to extract a robust pattern of neural activity involving the contra-lateral insular cortex, the thalamus, the brainstem and the cerebellum. No significant correlation with the temporal envelope of the nystagmus was found. The temporal analysis of the activation profiles highlighted a significantly longer duration of the evoked BOLD activity in the brainstem compared to the insular cortex suggesting a functional de-coupling between cortical and subcortical activity during the vestibular response.


Effect of growth factors on the proliferation and gene expression of human meibomian gland epithelial cells.
We hypothesize that growth factors, including epidermal growth factor (EGF) and bovine pituitary extract (BPE), induce proliferation, but not differentiation (e.g., lipid accumulation), of human meibomian gland epithelial cells. We also hypothesize that these actions involve a significant upregulation of genes linked to cell cycle processes, and a significant downregulation of genes associated with differentiation. Our objective was to test these hypotheses. Immortalized human meibomian gland and conjunctival epithelial cells were cultured for varying time periods in the presence or absence of EGF, BPE, EGF + BPE, or serum, followed by cell counting, neutral lipid staining, or RNA isolation for molecular biological procedures. Our studies show that growth factors stimulate a significant, time-dependent proliferation of human meibomian gland epithelial cells. These effects are associated with a significant upregulation of genes linked to cell cycle, DNA replication, ribosomes, and translation, and a significant decrease in those related to cell differentiation, tissue development, lipid metabolic processes, and peroxisome proliferator-activated receptor signaling. Serum-induced differentiation, but not growth factor-related proliferation, elicits a pronounced lipid accumulation in human meibomian gland epithelial cells. This lipogenic response is unique, and is not duplicated by human conjunctival epithelial cells. Our results demonstrate that EGF and BPE stimulate human meibomian gland epithelial cells to proliferate. Further, our findings show that action is associated with an upregulation of cell cycle and translation ontologies, and a downregulation of genetic pathways linked to differentiation and lipid biosynthesis.


Conserving plants within and beyond protected areas - still problematic and future uncertain.
Against a background of continuing loss of biodiversity, it is argued that for the successful conservation of threatened plant species we need to ensure the more effective integration of the various conservation actions employed, clarify the wording of the CBD targets and provide clearer operational guidance as to how they are to be implemented and their implementation monitored. The role and effectiveness of protected areas in conserving biodiversity and in particular plant species in situ are discussed as are recent proposals for a massive increase of their extent. The need for much greater effort and investment in the conservation or protection of threatened species outside protected areas where most plant diversity occurs is highlighted. The difficulties involved in implementing effective in situ conservation of plant diversity both at an area- and species/population-based level are discussed. The widespread neglect of species recovery for plants is noted and the desirability of making a clearer distinction between species recovery and reintroduction is emphasized. Key messages from a global overview of species recovery are outlined and recommendations made, including the desirability of each country preparing a national species recovery strategy. The projected impacts of global change on protected areas and on species conservation and recovery, and ways of addressing them are discussed.


Risk factors for post-neonatal intensive care unit discharge mortality among extremely low birth weight infants.
The study goal was to evaluate maternal and neonatal risk factors associated with post-neonatal intensive care unit (NICU) discharge mortality among extremely low birth weight (ELBW) infants. This is a retrospective analysis of ELBW (<1000 g) and <27 weeks' gestational age infants born in the Eunice Kennedy Shriver National Institute of Child Health and Human Development Neonatal Research Network sites between January 2000 and June 2007. Infants were tracked until death or 18 to 22 months' corrected age. Infants who died between NICU discharge and the 18- to 22-month follow-up visit were classified as post-NICU discharge mortality (P-NDM). Association of maternal and infant risk factors with P-NDM was determined using logistic regression analysis. A prediction model with 6 significant predictors was developed and validated. There were 5364 infants who survived to NICU discharge; 557 (10%) infants were lost to follow-up, and 107 infants died following NICU discharge. P-NDM rate was 22.3 per 1000 ELBW infants. In the prediction model, African American race, unknown maternal health insurance, and hospital stay ≥ 120 days significantly increased risk, and maternal exposure to intrapartum antibiotics was associated with decreased risk of P-NDM. We identified African American race, unknown medical insurance, and prolonged NICU stay as risk factors associated with P-NDM among ELBW infants.


An in vitro analysis of the effects of various topical antimicrobial agents on methicillin-resistant and methicillin-sensitive strains of Staphylococcus aureus.
Infections of acute and chronic wounds have a substantial negative impact on patient outcomes. Because bacterial resistance to traditional antimicrobials continues to increase, an in vitro study was conducted to examine current sensitivities of various methicillin-resistant and methicillin-sensitive strains of Staphylococcus aureus (MRSA and MSSA) to commonly used topical antimicrobial agents. Using fresh cultures of eight strains of MRSA and MSSA, the area of the zone of inhibition produced by various topical antimicrobials, including an aminoglycoside antibiotic, monocarboxylic acid antibiotic, pleuromutilin antibiotic, triple antibiotic ointment, and petrolatum ointment, was examined. Six culture plates per antimicrobial were prepared using the Kirby Bauer method; soy blood culture plates were inoculated with the bacteria, incubated for 24 hours at 37˚ C, and their zones of inhibition measured and calculated. Data were analyzed using ANOVA testing. Mupirocin treatment was the most effective antimicrobial, with areas of inhibition ranging from 30.34 cm2 to 61.70 cm2 (P <0.05), as compared to the next most effective, retapamulin, with areas of inhibition ranging from 11.97 cm2 to 23.54 cm2. This study provides current scientific data to help the development of a thoughtful rationale for the use of topical antimicrobials in wounds. Additional in vivo studies to substantiate these findings are needed.


Spectroscopic signatures of topological and diatom-vacancy defects in single-walled carbon nanotubes.
The optical properties, including UV-vis spectra and resonance Raman profiles, of pristine and defected single-walled carbon nanotubes (SWCNTs) are computed using state-of-the-art time-dependent density functional theory (TDDFT) as implemented using the Liouville-Lanczos approach to linear-response TDDFT. The CNT defects were of the form of Stone-Wales and diatom-vacancies. Our results are in very good agreement with experimental results where defects were introduced into a part of defect-free CNTs. In particular, we show that the first and second π-π* excitation energies are barely shifted due to the defects and associated with a relatively small reduction in the maxima of the absorption bands. In contrast, the resonance Raman spectra show close to an order of magnitude reduction in intensities, offering a means to distinguish between pristine and defected SWCNTs even at low defect concentrations.


Protein-mediated RNA folding governs sequence-specific interactions between rotavirus genome segments.
Segmented RNA viruses are ubiquitous pathogens, which include influenza viruses and rotaviruses. A major challenge in understanding their assembly is the combinatorial problem of a non-random selection of a full genomic set of distinct RNAs. This process involves complex RNA-RNA and protein-RNA interactions, which are often obscured by non-specific binding at concentrations approaching in vivo assembly conditions. Here, we present direct experimental evidence of sequence-specific inter-segment interactions between rotavirus RNAs, taking place in a complex RNA- and protein-rich milieu. We show that binding of the rotavirus-encoded non-structural protein NSP2 to viral ssRNAs results in the remodeling of RNA, which is conducive to formation of stable inter-segment contacts. To identify the sites of these interactions, we have developed an RNA-RNA SELEX approach for mapping the sequences involved in inter-segment base-pairing. Our findings elucidate the molecular basis underlying inter-segment interactions in rotaviruses, paving the way for delineating similar RNA-RNA interactions that govern assembly of other segmented RNA viruses.


Comparing the Performance of the S-TOFHLA and NVS Among and Between English and Spanish Speakers.
Given the growing body of evidence demonstrating the significant implications of health literacy on a myriad of outcomes, researchers continue to incorporate health literacy metrics in studies. With this proliferation in measurement of health literacy in research, it has become increasingly important to understand how various health literacy tools perform in specific populations. Our objective was to compare the performance of two widely used tests, the Short Test of Functional Health Literacy in Adults (S-TOFHLA) and the Newest Vital Sign (NVS) among and between a sample of English and Spanish-speaking patients. Adults (N = 402) ages 50-75 years participating in a trial to promote colorectal cancer screening completed in-person interviews which included both measures of health literacy. In the full sample, the tests were moderately correlated (r = 0.69, p < .0001); however, there was a stronger correlation among those completing the test in Spanish (r = 0.83) as compared with English (r = 0.58, p < .0001). English speakers more often were categorized as having adequate literacy by the S-TOFHLA as compared with the NVS, whereas Spanish speakers scored consistently low on both instruments. These findings indicate that the categorization of participants into levels of literacy is likely to vary, depending on whether the NVS or S-TOFHLA is used for assessment, a factor which researchers should be aware of when selecting literacy assessments.


Direct binding of CEP85 to STIL ensures robust PLK4 activation and efficient centriole assembly.
Centrosomes are required for faithful chromosome segregation during mitosis. They are composed of a centriole pair that recruits and organizes the microtubule-nucleating pericentriolar material. Centriole duplication is tightly controlled in vivo and aberrations in this process are associated with several human diseases, including cancer and microcephaly. Although factors essential for centriole assembly, such as STIL and PLK4, have been identified, the underlying molecular mechanisms that drive this process are incompletely understood. Combining protein proximity mapping with high-resolution structural methods, we identify CEP85 as a centriole duplication factor that directly interacts with STIL through a highly conserved interaction interface involving a previously uncharacterised domain of STIL. Structure-guided mutational analyses in vivo demonstrate that this interaction is essential for efficient centriolar targeting of STIL, PLK4 activation and faithful daughter centriole assembly. Taken together, our results illuminate a molecular mechanism underpinning the spatiotemporal regulation of the early stages of centriole duplication.


Structural analysis of synthetic homo- and copolyesters by electrospray ionization on a fourier transform ion cyclotron resonance mass spectrometer
The molecular structure of a series of homo- and copolyesters was studied using sustained off-resonance irradiation collisionally activation dissociation on a Fourier transform ion cyclotron resonance mass spectrometer. Electrospray ionization was used as an ionization technique. The most important fragmentation pathways of the homopolyesters poly(dipropoxylated bisphenol-A/adipic acid) and poly(dipropoxylated bisphenol-A/isophthalic acid) were studied. Six different dissociation mechanisms were observed which are very similar to the mechanisms found to occur during pyrolysis of these compounds. Four of these mechanisms are a result of cleavages of the ester bond and the others are due to cleavages of the ether bond or bisphenol-A unit. Some of the fragments expected are not present in the spectrum, indicating that each fragment has a specific sodium affinity. Sequence-specific fragments of two of the three copolyester sequences that theoretically can exist were experimentally observed. Fragments that originate from the third sequence are not unique and can also be formed from other sequences. Therefore, it was not possible to determine the presence of the third sequence. Copyright 2000 John Wiley & Sons, Ltd.


Controlled Release of Dexamethasone From an Intravitreal Delivery System Using Porous Silicon Dioxide.
The current study aims to evaluate a porous silicon-based drug delivery system meant for sustained delivery of dexamethasone (Dex) to the vitreous and retina. Dexamethasone was grafted covalently into the pore walls of fully oxidized porous silicon particles (pSiO2-COO-Dex), which then was evaluated for the pharmacological effect of the payload on cultured ARPE19 cells before intravitreal injection. The Dex release profile was investigated in a custom designed dynamic dissolution chamber to mimic the turnover of vitreous fluid in rabbit eyes. Ocular safety, in vivo release, and pharmacodynamics were evaluated in rabbit eyes, and the human VEGF-induced rabbit retinal vascular permeability model. Loading efficiency of Dex was 69 ± 9 μg per 1 mg of the pSiO2-COO-Dex particles. Dynamic in vitro release demonstrated a sustained mode when compared to free Dex, with the drug half-life extended by 5 times. The released Dex was unaltered and biologically active. In vivo drug release in rabbit eyes revealed a mode similar to the release seen in vitro, with a vitreous half-life of 11 days. At 2 and 4 weeks after a single intravitreal injection of pSiO2-COO-Dex particles (mean 2.71 ± 0.47 mg), intravitreal 500 ng of VEGF did not induce significant retinal vessel dilation or fluorescein leakage, while these events were observed in the eyes injected with empty pSiO2 particles or with free Dex. The retinal vessel score from fluorescein angiography for the control eyes was double the score for the eyes injected with pSiO2-COO-Dex. No adverse reaction was observed for the eyes injected with drug-loaded pSi particles during the course of the study. The porous silicon-based Dex delivery system (pSiO2-COO-Dex) can be administered safely into vitreous without toxicity. Dex release from the porous silicon particles was sustained for 2 months and was effective against VEGF-induced retinal vessel reaction.


Tracking human migrations by the analysis of the distribution of HLA alleles, lineages and haplotypes in closed and open populations.
The human leucocyte antigen (HLA) system shows extensive variation in the number and function of loci and the number of alleles present at any one locus. Allele distribution has been analysed in many populations through the course of several decades, and the implementation of molecular typing has significantly increased the level of diversity revealing that many serotypes have multiple functional variants. While the degree of diversity in many populations is equivalent and may result from functional polymorphism(s) in peptide presentation, homogeneous and heterogeneous populations present contrasting numbers of alleles and lineages at the loci with high-density expression products. In spite of these differences, the homozygosity levels are comparable in almost all of them. The balanced distribution of HLA alleles is consistent with overdominant selection. The genetic distances between outbred populations correlate with their geographical locations; the formal genetic distance measurements are larger than expected between inbred populations in the same region. The latter present many unique alleles grouped in a few lineages consistent with limited founder polymorphism in which any novel allele may have been positively selected to enlarge the communal peptide-binding repertoire of a given population. On the other hand, it has been observed that some alleles are found in multiple populations with distinctive haplotypic associations suggesting that convergent evolution events may have taken place as well. It appears that the HLA system has been under strong selection, probably owing to its fundamental role in varying immune responses. Therefore, allelic diversity in HLA should be analysed in conjunction with other genetic markers to accurately track the migrations of modern humans.


Pyrimidine metabolism and sugar nucleotide synthesis in rat liver.
With radioactive precursors, the labelling kinetics of the soluble pyrimidine nucleotides and of RNA were measured in rat liver to determine the contribution of the metabolic flows through synthesis de novo and the salvage pathway. To separate and quantify all pyrimidine nucleotides, an h.p.l.c. technique was developed using anion-exchange chromatography and reversed-phase chromatography. The concentrations of cytidine nucleotides were in the range of 30-45 nmol/g wet weight, and the concentrations of the uridine phosphates and of the UDP-sugars were approx. 6 and 20 times higher respectively. After a single injection of [14C]orotic acid and of [3H]cytidine, the specific radioactivities were determined as a function of time. The 14C/3H ratio was calculated and gave a good indication of the involvement of the different flows. It could be concluded that UTP derived from synthesis de novo and from the salvage pathway is not completely mixed before being utilized. The flow of the salvage pathway is relatively more directed to RNA synthesis in the nucleus and that of synthesis de novo to cytoplasmic processes. For CTP it could also be concluded that the flow of the salvage pathway was relatively more directed to RNA synthesis in the nucleus. Because of the nuclear localization of the enzyme CMP-NeuAc (N-acetylneuraminate) synthase, special attention was paid to CMP-NeuAc. However, a conclusion about a location about the synthesis of CMP-NeuAc could not unequivocally be drawn, because of the small differences in 14C/3H ratio and the different values for the CDP-lipids.


Relationship between depression and health-related quality of life in older Korean patients with chronic obstructive pulmonary disease.
The aims are to determine the prevalence of depression and examine its impact on the health-related quality of life (HRQoL) among older Korean patients with chronic obstructive pulmonary disease (COPD). Realising the effects of depression on HRQoL, many studies have reported the prevalence of depression and its negative impacts on HRQoL in COPD patients. However, the majority of the studies were performed in developed countries, with very few studies occurring in developing or non-developed countries. Survey with the comparison of HRQoL between the COPD patients with and without depression. A total of 91 COPD patients completed the St. George respiratory questionnaire (SGRQ), Medical outcomes short form-36 (SF-36) and Beck depression inventory (BDI). The prevalence of depression was 15.4%. The SGRQ total and impact subscores were significantly worse in the patients with depression (p < 0.05). The physical functioning, bodily pain, vitality and general health dimensions from the SF-36 were statistically poor in the patients with depression (p < 0.05). The patients with severe and very severe lung function defects showed significantly worse total, activity and impact scores of the SGRQ and significantly poor physical functioning, role physical, bodily pain, social functioning, mental health, role emotional and general health scores on the SF-36 (p < 0.05). In South Korea, the prevalence of depression is considerable, and depression has a negative impact on the HRQoL among older COPD patients. The study results suggest that depression is prevalent in older COPD patients in South Korea. The presence of depression negatively impacts on the HRQoL of the COPD patients. Considering the importance of the relationship between psychological problems and HRQoL in caring for and promoting the welfare of COPD patients, this study provides fundamental information and a basis for further evaluation of this issue in developing and non-developed countries.


Management of flu-like syndrome with cetirizine in patients with relapsing-remitting multiple sclerosis during therapy with interferon beta: Results of a randomized, cross-over, placebo-controlled pilot study.
Flu-like syndrome (FLS) is a common adverse event experienced by patients with relapsing-remitting multiple sclerosis (RRMS) treated with interferon beta (IFNβ). FLS can lead to poor treatment adherence and early IFNβ discontinuation. The involvement of interleukin-6 (IL-6) in the occurrence of FLS has been suggested. We hypothesized that cetirizine, a second-generation histamine H1 receptor antagonist able to reduce the levels of IL-6, might improve IFNβ-induced FLS. We conducted a pilot, cross-over, randomized, placebo-controlled, double-blind study to evaluate the efficacy of cetirizine 10 mg added after each IFNβ injection to the standard of care for FLS (acetaminophen or nonsteroidal anti-inflammatory drugs) on FLS in patients with RRMS treated with IFNβ. Patients were randomized to two treatment sequences: 1) 4-week treatment with placebo added to the standard treatment for FLS, followed by 4-week treatment with cetirizine added to the standard of care, and 2) first addition of cetirizine, then of placebo. The primary efficacy endpoint was the mean change of FLS severity [11-point visual analog scale (VAS)] after 4 weeks of treatment within each sequence. Forty-five patients (71.1% female, mean age 39.1 years, mean time from RRMS diagnosis 5.8 years) were randomized to treatment sequences 1 and 2. The differences between cetirizine and placebo in the intensity of FLS were not statistically significant: total mean VAS scores at 4 hours from IFNβ injection were 3.57 and 3.42 for cetirizine and placebo, respectively (difference -0.15; 95% confidence interval: from -0.74 to 0.44; p = 0.6029). The two treatments were similar also with regard to other efficacy measures considered and to the safety/tolerability profile. The addition of cetirizine to the standard of care for IFNβ-induced FLS in patients with RRMS does not seem to provide significant benefits compared with placebo. Further effort is required to understand the mechanisms underlying IFNβ-induced FLS. EudraCT 2013-001055-12.


MATERNALLY EXPRESSED PAB C-TERMINAL, a novel imprinted gene in Arabidopsis, encodes the conserved C-terminal domain of polyadenylate binding proteins.
Parental imprinting is important for seed development, but few imprinted genes have been identified in plants. The four known imprinted genes in Arabidopsis thaliana encode transcriptional regulators. Here, we describe a novel imprinted gene, MATERNALLY EXPRESSED PAB C-TERMINAL (MPC), which encodes the C-terminal domain of poly(A) binding proteins (PABPs). PABPs play roles in mRNA stability and translation. MPC interacts with proteins that also interact with the C-terminal domain of typical PABPs, suggesting that MPC may regulate translation by modulating PABP activity. In the endosperm, MPC is expressed only from the maternal allele. Reduction of MPC expression affects seed development. In dna methyltransferase1 (met1) mutants, MPC is ectopically expressed, and the paternal allele is active in the endosperm. CGs in the 5' flanking region and gene body of MPC lose methylation in a met1 background. Both regions are required to confer imprinted reporter expression, suggesting that the gene body contains imprinting control region elements. In Arabidopsis, DEMETER (DME) activates expression of maternal alleles. MPC expression is reduced in flowers and seeds in a dme-4 mutant but only after fertilization in dme-1. We conclude that other factors along with DME promote MPC expression and that DME has indirect effects on imprinted gene expression in endosperm.


Cell cycle regulation of myosin-V by calcium/calmodulin-dependent protein kinase II.
Organelle transport by myosin-V is down-regulated during mitosis, presumably by myosin-V phosphorylation. We used mass spectrometry phosphopeptide mapping to show that the tail of myosin-V was phosphorylated in mitotic Xenopus egg extract on a single serine residue localized in the carboxyl-terminal organelle-binding domain. Phosphorylation resulted in the release of the motor from the organelle. The phosphorylation site matched the consensus sequence of calcium/calmodulin-dependent protein kinase II (CaMKII), and inhibitors of CaMKII prevented myosin-V release. The modulation of cargo binding by phosphorylation is likely to represent a general mechanism regulating organelle transport by myosin-V.


A glimpse into smartphone screen reader use among blind teenagers in rural Nepal.
Access to appropriate assistive technology is a challenge worldwide and especially in low GDP-per-capita countries. Nepal is one example of a country with several coinciding challenges: some claim having a high rate of blindness in the general population, a low-GDP-per capita and some studies claim it has a low literacy rate, especially in rural areas. Without appropriate assistive technology, some disabled youth may not get full access to education. To gain insight into assistive technology use in rural Nepal, five blind teenagers in a mixed secondary school with disabled and non-disabled students in rural Nepal were interviewed about their daily smartphone use. The results show that all the participants used screen readers on donated smartphones. None of the participants had received formal training in using smartphone screen readers and therefore lacked knowledge about basic and essential operational aspects of the devices as well as misguided expectations about the technology. One implication of the findings is that smartphone accessibility features training material needs to be made easily available to schools and all disabled youth worldwide, as smartphones are increasingly becoming available in low-income remote regions with low literacy rates. The built-in accessibility features of smartphones promise disabled youth a non-stigmatizing platform for social participation and access to the information society.Implications for RehabilitationThe built-in accessibility features of smartphones provide disabled youth a non-stigmatizing practical platform for social participation and access to education.Training material on how to use smartphone accessibility features needs to be made easily available to schools and all disabled youth worldwide.


Zygosity determination in hairless mice by PCR based on Hr(hr) gene analysis.
We analyzed the Hr gene of a hairless mouse strain of unknown origin (HR strain, http://animal.nibio.go.jp/e_hr.html) to determine whether the strain shares a mutation with other hairless strains, such as HRS/J and Skh:HR-1, both of which have an Hr(hr) allele. Using PCR with multiple pairs of primers designed to amplify multiple overlapping regions covering the entire Hr gene, we found an insertion mutation in intron 6 of mutant Hr genes in HR mice. The DNA sequence flanking the mutation indicated that the mutation in HR mice was the same as that of Hr(hr) in the HRS/J strain. Based on the sequence, we developed a genotyping method using PCR to determine zygosities. Three primers were designed: S776 (GGTCTCGCTGGTCCTTGA), S607 (TCTGGAACCAGAGTGACAGACAGCTA), and R850 (TGGGCCACCATGGCCAGATTTAACACA). The S776 and R850 primers detected the Hr(hr) allele (275-bp amplicon), and S607 and R850 identified the wild-type Hr allele (244-bp amplicon). Applying PCR using these three primers, we confirmed that it is possible to differentiate among homozygous Hr(hr) (longer amplicons only), homozygous wild-type Hr(shorter amplicons only), and heterozygous (both amplicons) in HR and Hos:HR-1 mice. Our genomic analysis indicated that the HR, HRS/J, and Hos:HR-1 strains, and possibly Skh:HR-1 (an ancestor of Hos:HR-1) strain share the same Hr(hr) gene mutation. Our genotyping method will facilitate further research using hairless mice, and especially immature mice, because pups can be genotyped before their phenotype (hair coat loss) appears at about 2 weeks of age.


Vaccinia virus intracellular mature virions contain only one lipid membrane.
Vaccinia virus (VV) morphogenesis commences with the formation of lipid crescents that grow into spherical immature virus (IV) and then infectious intracellular mature virus (IMV) particles. Early studies proposed that the lipid crescents were synthesized de novo and matured into IMV particles that contained a single lipid bilayer (S. Dales and E. H. Mosbach, Virology 35:564-583, 1968), but a more recent study reported that the lipid crescent was derived from membranes of the intermediate compartment (IC) and contained a double lipid bilayer (B. Sodiek et al., J. Cell Biol. 121:521-541, 1993). In the present study, we used high-resolution electron microscopy to reinvestigate the structures of the lipid crescents, IV, and IMV particles in order to determine if they contain one or two membranes. Examination of thin sections of Epon-embedded, VV-infected cells by use of a high-angular-tilt series of single sections, serial-section analysis, and high-resolution digital-image analysis detected only a single, 5-nm-thick lipid bilayer in virus crescents, IV, and IMV particles that is covered by a 8-nm-thick protein coat. In contrast, it was possible to discern tightly apposed cellular membranes, each 5 nm thick, in junctions between cells and in the myelin sheath of Schwann cells around neurons. Serial-section analysis and angular tilt analysis of sections detected no continuity between virus lipid crescents or IV particles and cellular membrane cisternae. Moreover, crescents were found to form at sites remote from IC membranes-namely, within the center of virus factories and within the nucleus-demonstrating that crescent formation can occur independently of IC membranes. These data leave unexplained the mechanism of single-membrane formation, but they have important implications with regard to the mechanism of entry of IMV and extracellular enveloped virus into cells; topologically, a one-to-one membrane fusion suffices for delivery of the IMV core into the cytoplasm. Consistent with this, we have demonstrated previously by confocal microscopy that uncoated virus cores within the cytoplasm lack the IMV surface protein D8L, and we show here that intracellular cores lack the surface protein coat and lipid membrane.


L1 cell adhesion molecule overexpression in hepatocellular carcinoma associates with advanced tumor progression and poor patient survival.
L1 cell adhesion molecule (L1CAM), as a member of the immunoglobulin superfamily, has recently been observed in a variety of human malignancies. However, no data of L1CAM are available for hepatocellular carcinoma (HCC). The aim of this study was to investigate the expression of L1CAM in HCC and determine its correlation with tumor progression and prognosis. One-hundred and thirty HCC patients who had undergone curative liver resection were selected and immunohistochemistry, Western blotting, and quantitative real time polymerase chain reaction (Q-PCR) were performed to analyze L1CAM expression in the respective tumors. Immunohistochemistry, Western blotting, and Q-PCR consistently confirmed the overexpression of L1CAM in HCC tissues compared with their adjacent nonneoplastic tissues at both protein and gene level (both P <0.01). Additionally, the high expression of L1CAM was significantly associated with advanced tumor stage (P = 0.02) and advanced tumor grade (P = 0.03), respectively. Moreover, HCC patients with high L1CAM expression were significantly associated with lower 5-year overall survival (P <0.01) and lower 5-year disease-free survival (P <0.01), respectively. The Cox proportional hazards model further showed that L1CAM over-expression was an independent poor prognostic factor for both 5-year disease-free survival (P = 0.02) and 5-year overall survival (P = 0.008) in HCC. Our data suggest for the first time that L1CAM expression in HCC was significantly correlated with the advanced tumor progression and was an independent poor prognostic factor for both overall survival and disease-free survival in patients with HCC. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1970024872761542.


High resolution FTIR imaging provides automated discrimination and detection of single malaria parasite infected erythrocytes on glass.
New highly sensitive tools for malaria diagnostics are urgently needed to enable the detection of infection in asymptomatic carriers and patients with low parasitemia. In pursuit of a highly sensitive diagnostic tool that can identify parasite infections at the single cell level, we have been exploring Fourier transform infrared (FTIR) microscopy using a Focal Plane Array (FPA) imaging detector. Here we report for the first time the application of a new optic configuration developed by Agilent that incorporates 25× condenser and objective Cassegrain optics with a high numerical aperture (NA = 0.81) along with additional high magnification optics within the microscope to provide 0.66 micron pixel resolution (total IR system magnification of 61×) to diagnose malaria parasites at the single cell level on a conventional glass microscope slide. The high quality images clearly resolve the parasite's digestive vacuole demonstrating sub-cellular resolution using this approach. Moreover, we have developed an algorithm that first detects the cells in the infrared image, and secondly extracts the average spectrum. The average spectrum is then run through a model based on Partial Least Squares-Discriminant Analysis (PLS-DA), which diagnoses unequivocally the infected from normal cells. The high quality images, and the fact this measurement can be achieved without a synchrotron source on a conventional glass slide, shows promise as a potential gold standard for malaria detection at the single cell level.


Conscious sedation and analgesia with rectal ketamine in the Macaca fuscata monkey.
Conscious sedation is commonly utilized in pediatric dentistry. Although opioid analgesics are often employed, patient safety would be enhanced if nonopioid drugs were used. The purpose of this study was to determine if rectal ketamine could produce plasma concentrations that would achieve both conscious sedation and analgesia to gingival needle puncture. Five 2-year-old male Macaca fuscata monkeys were given rectal ketamine at a dosage of 60 mg/kg and 90 mg/kg one week apart. Blood was drawn at selected times after administration, and vital signs, level of sedation, and consciousness were assessed. Plasma ketamine concentrations ranged from 240 to 820 ng/mL and from 390 to 3120 ng/mL after rectal administration at doses of 60 mg/kg and 90 mg/kg, respectively. Two monkeys after the high dose showed analgesia to a gingival needle puncture at plasma ketamine concentrations that ranged from 1390 to 3120 ng/mL. A good level of sedation was consistently observed in four monkeys (80%) following rectal ketamine at a dosage of 90 mg/kg, whereas one monkey showed a consistently good level at a dosage of 60 mg/kg. Sedation and dose were significantly (P less than 0.001) associated with plasma ketamine concentrations; physiologic parameters were not (P greater than 0.05). The results of this study suggest that rectal ketamine can produce plasma concentrations of the drug sufficient to achieve sedation in the monkey. The attainment of concomitant analgesia to a gingival needle puncture was not as predictable.


Modulation of Shigella virulence in response to available oxygen in vivo.
Bacteria coordinate expression of virulence determinants in response to localized microenvironments in their hosts. Here we show that Shigella flexneri, which causes dysentery, encounters varying oxygen concentrations in the gastrointestinal tract, which govern activity of its type three secretion system (T3SS). The T3SS is essential for cell invasion and virulence. In anaerobic environments (for example, the gastrointestinal tract lumen), Shigella is primed for invasion and expresses extended T3SS needles while reducing Ipa (invasion plasmid antigen) effector secretion. This is mediated by FNR (fumarate and nitrate reduction), a regulator of anaerobic metabolism that represses transcription of spa32 and spa33, virulence genes that regulate secretion through the T3SS. We demonstrate there is a zone of relative oxygenation adjacent to the gastrointestinal tract mucosa, caused by diffusion from the capillary network at the tips of villi. This would reverse the anaerobic block of Ipa secretion, allowing T3SS activation at its precise site of action, enhancing invasion and virulence.


Background Reduction in STED-FCS Using a Bivortex Phase Mask.
Fluorescence correlation spectroscopy (FCS) is a valuable tool to study the molecular dynamics in living cells. When used together with a super-resolution stimulated emission depletion (STED) microscope, STED-FCS can measure diffusion processes on the nanoscale in living cells. In two-dimensional (2D) systems like the cellular plasma membrane, a ring-shaped depletion focus is most commonly used to increase the lateral resolution, leading to more than 25-fold decrease in the observation volume, reaching the relevant scale of supramolecular arrangements. However, STED-FCS faces severe limitations when measuring diffusion in three dimensions (3D), largely due to the spurious background contributions from undepleted areas of the excitation focus that reduce the signal quality and ultimately limit the resolution. In this paper, we investigate how different STED confinement modes can mitigate this issue. By simulations as well as experiments with fluorescent probes in solution and in cells, we demonstrate that the coherent-hybrid (CH) depletion pattern created by a bivortex phase mask reduces background most efficiently and thus provides superior signal quality under comparable reduction of the observation volume. Featuring also the highest robustness to common optical aberrations, CH-STED can be considered the method of choice for reliable STED-FCS-based investigations of 3D diffusion on the subdiffraction scale.


Retrograde transpopliteal access is safe and effective-it should be added to the vascular surgeon's portfolio.
The aim of the study was to review the outcomes of superficial femoral artery (SFA) interventions using a retrograde transpopliteal access approach after failed antegrade recanalization. A database of patients undergoing endovascular treatment of the SFA between 2008 and 2011 was retrospectively queried, and those cases with transpopliteal artery retrograde access were analyzed. Time-dependent outcomes were determined by Kaplan-Meier survival analyses. A total of 16 patients (75% men; mean age 61 ± 9 years) underwent retrograde popliteal access after failed antegrade attempts. Patients had multiple cardiovascular comorbidities with a mean modified cardiac index score of 3.1 ± 1.8. The reason for intervention was lifestyle-limiting claudication in 67% of cases and critical ischemia in the remainder. Most of the lesions were Trans-Atlantic Inter-Society Consensus II C and D. Retrograde ultrasound-guided puncture of the popliteal artery was successful in all cases and there were no local site complications. Intervention was successful in 94% of cases. One uncomplicated perforation (7%) was encountered during attempted recanalization of the SFA in the thigh. There was no perioperative morbidity or 30-day mortality. The 30-day major adverse cardiovascular events rate was 6% but both 30-day major adverse limb events and the 30-day major amputation rate were 0%. There was a 40% increase in actual ankle-brachial index (ABI); 93% of patients achieved an ABI rise >0.15. On longer term follow-up, 2 patients developed restenosis and 1 an asymptomatic occlusion. Both restenosis patients required re-angioplasty. Two patients required expected toe amputations as a result of their presenting symptoms. The primary patency was 66 ± 9%, assisted patency 81 ± 9%, and secondary patency 87 ± 8% at 2 years. Limb salvage was 100%. Clinical efficacy was 63 ± 9% at 2 years. Ultrasound-guided retrograde transpopliteal access is a safe and successful technique, which extends the ability to perform endovascular interventions after failed antegrade approaches.


Bronchial reactivity to inhaled histamine and annual rate of decline in FEV1 in male smokers and ex-smokers.
We examined the relations between bronchial reactivity, baseline FEV1, and annual decline of height corrected FEV1 (delta FEV1/ht3) over 7.5 years in 227 men (117 smokers, 71 ex-smokers, and 39 non-smokers). Men with a clinical diagnosis of asthma or receiving bronchodilator treatment were excluded. Bronchial reactivity was determined as the provocation concentration (PC20) of inhaled histamine sufficient to reduce FEV1 by 20%; subjects were divided into reactors (PC20 less than or equal to 16 mg/ml) and non-reactors (PC20 greater than 16 mg/ml). Thirty per cent of smokers, 24% of ex-smokers, and 5% of non-smokers were reactors. When smokers who were reactors were compared with non-reactors, the reactors showed a lower baseline FEV1 as percentage predicted in 1981-2 (85% v 108%), and a faster delta FEV1/ht3 (14.1 v 9.2 ml/y/m3). Baseline FEV1 correlated with PC20 in both smokers (rs = 0.51) and ex-smokers (rs = 0.61), and all 15 subjects with an FEV1 under 80% of the predicted value were reactors. In ex-smokers delta FEV1/ht3 was similar in reactors and non-reactors (m 9.0 v 7.4 ml/y/m3), despite significant differences in baseline FEV1. When analysis was confined to men with a baseline FEV1 over 80% predicted, the prevalence of reactors was significantly increased among smokers and slightly increased among ex-smokers compared with non-smokers, though the mean FEV1 was higher in the non-smokers. Bronchial reactivity was not increased in smokers aged 35 years or less. In smokers delta FEV1/ht3 was faster in those with a personal history of allergy (usually allergic rhinitis), but was not related to a family history of allergic disease, total serum immunoglobulin E level, absolute blood eosinophil count, or skinprick test score. delta FEV1/ht3 was also faster in all subjects taking beta blocker drugs. Thus increased bronchial reactivity was associated with accelerated decline of FEV1 in smokers. Although the association could be a consequence of a lower lower baseline FEV1, a trend towards increased reactivity was found in smokers with normal baseline FEV1 and delta FEV1/ht3 was dissociated from increased reactivity in ex-smokers. These findings are compatible with the "Dutch hypothesis," but the association between allergic features and accelerated delta FEV1/ht3 was relatively weak, and increased reactivity may follow rather than precede the onset of smoking.


First Report of Soybean Neocosmospora Stem Rot Caused by Neocosmospora vasinfecta var. vasinfecta in China.
During October 2009, the occurrence of a disease on soybean (Glycine max) was observed in several fields in Boluo County and Zengcheng City, Guangdong Province. Top leaves of infected plants initially turned yellow and plants eventually were defoliated, while stems and roots became black and rotted. The stem lesions sometimes extended 10 to 15 cm upward from the soil surface. Orange-to-brown spherical fruiting bodies, which were very similar with those of the soybean red crown rot pathogen, scattered or congregated on the stem lesions. Plants with symptoms were sampled from fields. Fruiting bodies were excised from diseased tissues. Microscopic examination revealed that they were perithecia, globose to pyriform, ostiolate with a short neck, and measured 160 to 298 × 151 to 235 μm. Under gentle pressure, asci and ascospores were exuded from these perithecia. Asci were eight spored, narrowly cylindrical to clavate, thin walled, with a short stalk, and measured 58 to 124 μm long and 8 to 15 μm in diameter. Ascospores were uniseriately arranged, globose to ellipsoid, thick walled, one celled, hyaline to pale, and measured 14 to 17 × 8 to 12 μm. Isolation was made from stem tissues at the edge of disease lesions on potato dextrose agar (PDA) amended with streptomycin sulfate and incubated at 25°C. Mycelia were white and floccose. Conidia were cylindrical to oblong-ellipsoidal, hyaline, one celled, and measured 6 to 22 × 2 to 5 μm, aggregating in a slimy mass on the apex of the conidiogenous cell. Abundant orange-to-brown spherical perithecia were produced on the colony. Ascospores had walls with a rugose ornamentation that could be clearly seen under a scanning electron microscope. The fungus was identified as Neocosmospora vasinfecta var. vasinfecta (anamorph Acremonium sp.) (1). The internal transcribed spacer (ITS) region of rDNA of two isolates were amplified with universal primers ITS1/ITS4 and sequenced (GenBank Accession No. JF705861 and JF705862), and comparisons with GenBank accessions showed 99% similarity with N. vasinfecta strain Pec070 (Accession No. FJ940902) and strain NRRL22497 (Accession No. AY381142). Pathogenicity tests were conducted. Five, 3-week-old seedlings of soybean cv. Huaxia No. 3 planted in plastic pots (20 cm in diameter) were wounded with a needle at the base of the stem below the soil line and near the root system, and then inoculated by drenching the soil with a conidial suspension (105 per ml). Control plants were inoculated with sterile water. There were six replicates for each treatment. The treated plants were incubated at 25 ± 2°C in a greenhouse. All inoculated plants exhibited symptoms of leaf yellowing and black rot of stems and roots 3 weeks after inoculation. N. vasinfecta var. vasinfecta was reisolated from the diseased plants. All control plants remained healthy. To our knowledge, this is the first observation of Neocosmospora stem rot of soybean in China. The pathogen could pose a threat to soybean, which is a major crop in China. This disease has been previously reported in the United States though the anamorph of the pathogen has either not been identified or has been identified as a Cylindrocarpon sp. (2,4). This fungus is also associated with human infections (3). References: (1) P. F. Cannon and D. L. Hawksworth. Trans. Br. Mycol. Soc. 82:673, 1984. (2) F. A. Gray et al. Plant Dis. 64:321, 1980. (3) P. Manikandan et al. Med. Mycol. 46:279, 2008. (4) D. V. Phillips. Phytopathology 62:612, 1972.


Cryoprotection and banking of living cells in a 3D multiple emulsion-based carrier.
The ability to preserve stem cells/cells with minimal damage for short and long periods of time is essential for advancements in biomedical therapies and biotechnology. New methods of cell banking are continuously needed to provide effective damage prevention to cells. This paper puts forward a solution to the problem of the low viability of cells during cryopreservation in a traditional suspension and storage by developing innovative multiple emulsion-based carriers for the encapsulation and cryopreservation of cells. During freezing-thawing processes, irreversible damage to cells occurs as a result of the formation of ice crystals, cell dehydration, and the toxicity of cryoprotectant. The proposed method was effective due to the "flexible" protective structure of multiple emulsions, which was proven by a high cell survival rate, above 90%. Results make new contributions in the fields of cell engineering and biotechnology and contribute to the development of methods for banking biological material.


Spray-Dried Hierarchical Aggregates of Iron Oxide Nanoparticles and Their Functionalization for Downstream Processing in Biotechnology.
In this work, the structuring of iron oxide nanoparticles via spray-drying (SD) of aqueous suspensions is investigated, leading to micrometer-sized aggregates with saturation magnetization comparable to that of the individual nanoparticles. Interestingly, the superparamagnetic behavior is retained despite the multicore structure. Modification of the aggregates via the addition of silica nanoparticles to the suspension allows for control of the resulting magnetization by adjusting the iron oxide content. Moreover, the morphology of the produced aggregates is gradually shifted from irregular inflated-like shapes in case of pure iron oxide aggregates to reach spherical structures when bringing the silica content to only 20%. The aggregates with different magnetization can be effectively separated in a simple column with an attached permanent magnet. Functionalization of pure iron oxide aggregates with a previously coupled ligand holding a nitrilotriacetic acid (NTA)-like moiety and subsequent loading with Ni2+ ions leads to the ability to bind 6-histidine (His6)-tagged target proteins via chelation complexes for magnetic separation. The application of the presented system for the purification of recombinant protein A in multiple cycles is shown. The recyclability of the separation system in combination with the high degree of magnetic separation is promising for future applications in the field of preparative in situ protein purification.


Hospitalization of abused and neglected children.
To describe the use of inpatient hospitalization for abused and neglected children living in a metropolitan area. Retrospective record review of abused and neglected children admitted in 1992 and 1993 to hospitals with 87% of metropolitan area pediatric admissions; comparison of these data with population, crisis nursery, and child protective services data. Thirty-four abused and neglected children were admitted to hospital, representing 0.3% (34/11,066; 95% confidence interval, 0%-1.2%) of pediatric admissions and 0.2% (34/19,950; 95% confidence interval, 0%-0.6%) of child protective services reports. This represents a rate of hospitalization for child abuse of 10 children (95% confidence interval, 0-46) per 100,000 child population per year. Seven hundred fifteen children were admitted to the crisis nursery by child protective services. Of those admitted to the hospital, 12 needed intensive care, 5 of whom died. Only 3 of 34 hospital-admitted children had private health insurance; 19 of 34 were younger than 1 year. Inpatient hospitalization for abuse represented a small fraction of total pediatric admissions and of child protective services reports. Comprehensive medical care for most abused children and medical education about child abuse must occur in outpatient settings.


Single Nucleotide Polymorphisms in Regulator-Encoding Genes Have an Additive Effect on Virulence Gene Expression in a Vibrio cholerae Clinical Isolate.
Vibrio cholerae is the etiological agent of the infectious disease cholera, which is characterized by vomiting and severe watery diarrhea. Recently, V. cholerae clinical isolates have demonstrated increased virulence capabilities, causing more severe symptoms with a much higher rate of disease progression than previously observed. We have identified single nucleotide polymorphisms (SNPs) in four virulence-regulatory genes (hapR, hns, luxO, and vieA) of a hypervirulent V. cholerae clinical isolate, MQ1795. Herein, all SNPs and SNP combinations of interest were introduced into the prototypical El Tor reference strain N16961, and the effects on the production of numerous virulence-related factors, including cholera toxin (CT), the toxin-coregulated pilus (TCP), and ToxT, were analyzed. Our data show that triple-SNP (hapR hns luxO and hns luxO vieA) and quadruple-SNP combinations produced the greatest increases in CT, TCP, and ToxT production. The hns and hns luxO SNP combinations were sufficient for increased TCP and ToxT production. Notably, the hns luxO vieA triple-SNP combination strain produced TCP and ToxT levels similar to those of MQ1795. Certain SNP combinations (hapR and hapR vieA) had the opposite effect on CT, TCP, and ToxT expression. Interestingly, the hns vieA double-SNP combination strain increased TCP production while decreasing CT production. Our findings suggest that SNPs identified in the four regulatory genes, in various combinations, are associated with increased virulence capabilities observed in V. cholerae clinical isolates. These studies provide insight into the evolution of highly virulent strains. IMPORTANCE Cholera, an infectious disease of the small intestine caused by the aquatic bacterium Vibrio cholerae, often results in vomiting and acute watery diarrhea. If left untreated or if the response is too slow, the symptoms can quickly lead to extreme dehydration and ultimately death of the patient. Recent anecdotal evidence of cholera patients suffering from increasingly severe symptoms and of disease progression at a much higher rate than previously observed has emerged. As recent cholera outbreaks caused by increasingly virulent strains have resulted in higher mortality rates, the need to investigate the mechanism(s) allowing this observed increased virulence is apparent. The significance of our research is in identifying the mechanism for increased virulence capabilities, which will allow the development of a model that will greatly enhance our understanding of cholera disease and V. cholerae pathogenesis, leading to broader biomedical impacts, as cholera serves as a model for other enteric diarrheal diseases.