A novel derivative of the natural agent deguelin for cancer chemoprevention and therapy.
The natural compound deguelin has promising preventive and therapeutic activity against diverse cancers by directly binding to heat shock protein-90 and thus suppressing its function. Potential side effects of deguelin over a certain dose, however, could be a substantial obstacle to its clinical use. To develop a derivative(s) of deguelin with reduced potential side effects, we synthesized five deguelin analogues (SH-02, SH-03, SH-09, SH-14, and SH-15) and compared them with the parent compound and each other for structural and biochemical features; solubility; and antiproliferative effects on normal, premalignant, and malignant human bronchial epithelial (HBE) and non-small-cell lung cancer (NSCLC) cell lines. Four derivatives destabilized hypoxia-inducible factor-1alpha as potently as did deguelin. Reverse-phase protein array (RPPA) analysis in H460 NSCLC cells revealed that deguelin and the derivatives suppressed expression of a number of proteins including heat shock protein-90 clients and proteins involved in the phosphoinositide 3-kinase/Akt pathway. One derivative, SH-14, showed several features of potential superiority for clinical use: the highest apoptotic activity; no detectable influence on Src/signal transducer and activator of transcription signaling, which can promote cancer progression and is closely related to pathogenesis of Parkinson's disease (deguelin, SH-02 and SH-03 strongly activated this signaling); better aqueous solubility; and less cytotoxicity to immortalized HBE cells (versus deguelin) at a dose (1 micromol/L) that induced apoptotic activity in most premalignant and malignant HBE and NSCLC cell lines. These collective results suggest that the novel derivative SH-14 has strong potential for cancer chemoprevention and therapy, with equivalent efficacy and lesser toxicity (versus deguelin).


Determinants of biosecurity behaviour of British cattle and sheep farmers-a behavioural economics analysis.
The paper analyses the impact of a priori determinants of biosecurity behaviour of farmers in Great Britain. We use a dataset collected through a stratified telephone survey of 900 cattle and sheep farmers in Great Britain (400 in England and a further 250 in Wales and Scotland respectively) which took place between 25 March 2010 and 18 June 2010. The survey was stratified by farm type, farm size and region. To test the influence of a priori determinants on biosecurity behaviour we used a behavioural economics method, structural equation modelling (SEM) with observed and latent variables. SEM is a statistical technique for testing and estimating causal relationships amongst variables, some of which may be latent using a combination of statistical data and qualitative causal assumptions. Thirteen latent variables were identified and extracted, expressing the behaviour and the underlying determining factors. The variables were: experience, economic factors, organic certification of farm, membership in a cattle/sheep health scheme, perceived usefulness of biosecurity information sources, knowledge about biosecurity measures, perceived importance of specific biosecurity strategies, perceived effect (on farm business in the past five years) of welfare/health regulation, perceived effect of severe outbreaks of animal diseases, attitudes towards livestock biosecurity, attitudes towards animal welfare, influence on decision to apply biosecurity measures and biosecurity behaviour. The SEM model applied on the Great Britain sample has an adequate fit according to the measures of absolute, incremental and parsimonious fit. The results suggest that farmers' perceived importance of specific biosecurity strategies, organic certification of farm, knowledge about biosecurity measures, attitudes towards animal welfare, perceived usefulness of biosecurity information sources, perceived effect on business during the past five years of severe outbreaks of animal diseases, membership in a cattle/sheep health scheme, attitudes towards livestock biosecurity, influence on decision to apply biosecurity measures, experience and economic factors are significantly influencing behaviour (overall explaining 64% of the variance in behaviour). Three other models were run for the individual regions (England, Scotland and Wales). A smaller number of variables were included in each model to account for the smaller sample sizes. Results show lower but still high levels of variance explained for the individual models (about 40% for each country). The individual models' results are consistent with those of the total sample model. The results might suggest that ways to achieve behavioural change could include ensuring increased access of farmers to biosecurity information and advice sources.


Interactions between Soluble Species of β-Amyloid and α-Synuclein Promote Oligomerization while Inhibiting Fibrillization.
Aggregations of β-amyloid (Aβ) and α-synuclein (αS) into oligomeric and fibrillar assemblies are the pathological hallmarks of Alzheimer's and Parkinson's diseases, respectively. Although Aβ and αS affect different regions of the brain and are separated at the cellular level, there is evidence of their eventual interaction in the pathology of both disorders. Characterization of interactions of Aβ and αS at various stages of their aggregation pathways could reveal mechanisms and therapeutic targets for the prevention and cure of these neurodegenerative diseases. In this study, we comprehensively examined the interactions and their molecular manifestations using an array of characterization tools. We show for the first time that αS monomers and oligomers, but not αS fibrils, inhibit Aβ fibrillization while promoting oligomerization of Aβ monomers and stabilizing preformed Aβ oligomers via coassembly, as judged by Thioflavin T fluorescence, transmission electron microscopy, and SDS- and native-PAGE with fluorescently labeled peptides/proteins. In contrast, soluble Aβ species, such as monomers and oligomers, aggregate into fibrils, when incubated alone under the otherwise same condition. Our study provides evidence that the interactions with αS soluble species, responsible for the effects, are mediated primarily by the C-terminus of Aβ, when judged by competitive immunoassays using antibodies recognizing various fragments of Aβ. We also show that the C-terminus of Aβ is a primary site for its interaction with αS fibrils. Collectively, these data demonstrate aggregation state-specific interactions between αS and Aβ and offer insight into a molecular basis of synergistic biological effects between the two polypeptides.


Sex pheromone of the queen butterfly: biology.
Males of the queen butterfly Danaus gilippus berenice, deprived of the two extrusible brushlike "hairpencils" at the rear of their abdomen, are capable of courting females but incapable of seducing them. In normal courtship, an aphrodisiac secretion associated with the hairpencils is transferred by way of tiny cuticular "dust" particles to the antennae of the females. Of the two substances identified from the secretion, one (the ketone) acts as the chemical messenger that induces the females to mate. The only known function of the other compound (the diol) is to serve as a glue that sticks the dust to the female. Males were reared under conditions in which they produced subnormal amounts of ketone and showed reduced seductive capacity. Under certain experimental circumstances, the competence of these males was restored by addition of synthetic ketone.


Cellulose microfibrils: visualization of biosynthetic and orienting complexes in association with the plasma membrane.
Cellulose microfibril biosynthesis, assembly, and orientation in the unicellular green alga, Oocystis, is visualized in association with a linear enzyme complex embedded in the B face of the plasma membrane. Granule bands of the A face and complementary ridges of the B face are postulated to assist in the orientation of recently synthesized microfibrils. A model for microfibril synthesis and orientation is proposed and correlated with current hypotheses regarding cellulose biosynthesis in higher plants.


Invasive black spiny-tailed iguanas (Ctenosaura similis) on Gasparilla Island, Florida, USA.
The native range of Ctenosaura similis extends from southern Mexico through Panama. From an initial introduction of 3 animals in 1979, the species now numbers in the thousands on Gasparilla Island in southwest Florida. In response to complaints of property damage from residents and threats to native species, local officials and the US Department of Agriculture Wildlife Services began a removal program in 2008. Through 2011, trappers removed 9467 ctenosaurs. The number removed declined from 32 iguanas/day in 2008 to 1.9 iguanas/day in 2011 despite no easing of the control effort. We necropsied 2757 ctenosaurs to document aspects of their natural history. Females outnumbered males overall, although the largest size class (>300 mm snout-vent length) included 32 males and just 2 females. Reproduction was seasonal. We found oviducal eggs in females from early Apr to early Jun, approximately 2 months later than C. similis in its native range. We trapped hatchlings from late Jul to early Oct coincident with the summer rainy season. Clutch size increased with female body size, with 62 being the largest clutch size recorded. In general, the biology of the invasive population on Gasparilla Island resembles native C. similis populations in Central America, except for the lack of large individuals. We suggest that shorter day length and colder temperatures create environmental conditions that are suboptimal for individual growth compared to those in the native range.


Pathogenesis of skin ulcers: lessons from the Mycobacterium ulcerans and Leishmania spp. pathogens.
Skin ulcers are most commonly due to circulatory or metabolic disorders and are a major public health concern. In developed countries, chronic wounds affect more than 1 % of the population and their incidence is expected to follow those observed for diabetes and obesity. In tropical and subtropical countries, an additional issue is the occurrence of ulcers of infectious origins with diverse etiologies. While the severity of cutaneous Leishmaniasis correlates with protective immune responses, Buruli ulcers caused by Mycobacterium ulcerans develop in the absence of major inflammation. Based on these two examples, this review aims to demonstrate how studies on microorganism-provoked wounds can provide insight into the molecular mechanisms controlling skin integrity. We highlight the potential interest of a mouse model of non-inflammatory skin ulceration caused by intradermal injection of mycolactone, an original lipid toxin with ulcerative and immunosuppressive properties produced by M. ulcerans.


Is it possible to extrapolate the rates of resistance of Escherichia coli from asymptomatic bacteriuria in pregnant women to those of E. coli in uncomplicated community-acquired UTI?
Treatment of uncomplicated urinary tract infections in primary care is generally empirical without requesting urine culture and based on biased resistance data collected from selected patients, most of them having risk factors for the isolation of resistant microorganisms. In order to overcome the lack of information on the real resistance rates in uncomplicated UTI, we compared antimicrobial phenotype and genotype of Escherichia coli isolated from pregnant women with asymptomatic bacteriuria (culture always performed) with those from women with uncomplicated acute cystitis (culture rarely performed) of different age groups. Between September 2017 and March 2018, 103 urines were randomly collected from pregnant women aged between 16 and 47 with asymptomatic bacteriuria (AB) (n=42), not hospitalized women in the same age range with uncomplicated acute cystitis (UAC) (n=31) and women older than 47 not hospitalized with UAC (n=30). Bacteria identification was performed using mass spectrometry and the antibiogram by broth microdilution. Genetic typification was carried out by pulsed-field gel electrophoresis. There are no significant differences between the groups of patients in the antibiotic susceptibility. Likewise, as expected, a wide genetic diversity is observed among the strains of E. coli studied without significant differences between the three groups. We propose a simple model that could provide better guidance for selection of empirical antimicrobial therapy for non-pregnant women with UAC than do generic hospital antibiogram data based on reliably extrapolating the susceptibility data of strains isolated from pregnant women with AB as representation of women with community-acquired UAC.


[Verotoxin-producing Escherichia coli: quantitative model of exposure and risk scenarios in cattle carcasses in Argentina].
Quantify contamination by verotoxin-producing Escherichia coli associated with hemolytic uremic syndrome (VTEC-HUS) in cattle carcasses and generate estimates of exposure in three likely scenarios. A model was constructed of the frequency and magnitude of VTEC-HUS contamination from primary production to the removal of the carcasses from cold storage, based on the published scientific information, epidemiological data, and information from local experts. The probability distributions that best described each step in the process and scenarios were input to the @Risk program with multiple simulations using Monte Carlo analysis. Pearson s correlation test was used for the sensitivity analysis. The estimated frequency of carcasses with VTEC-HUS was 0.37 (95% CI: 0.26 to 0.58) and the final load of VTEC-HUS was 0.47 log CFU/carcass (95% CI: -2.46 to 3.62). The most closely related variables were the fattening system (r = -0.681) and the theoretical concentration of VTEC-HUS on the cattle's skin (r = 0.702). Vaccinating the animals reduced the frequency of VTEC-HUS in the carcasses by 54.1%, although there were no significant changes in the final VTEC-HUS load. Washing the carcasses reduced the final load by 0.42 log CFU/carcass compared with the baseline model, without any change in the frequency. A 50%-60% increase in the percentage of animals fattened in pens would increase the frequency of carcasses contaminated with VTEC-HUS by 15%-23%. Vaccinating the animals was the most effective scenario for reducing introduction of the bacteria in the beef production chain. Intensifying livestock production will increase the public health risk due to greater exposure to VTEC-HUS.


Development of postural control in healthy children: a functional approach.
From a set of experimental studies showing how intersegmental coordination develops during childhood in various posturokinetic tasks, we have established a repertoire of equilibrium strategies in the course of ontogenesis. The experimental data demonstrate that the first reference frame used for the organization of balance control during locomotion is the pelvis, especially in young children. Head stabilization during posturokinetic activities, particularly locomotion, constitutes a complex motor skill requiring a long time to develop during childhood. When studying the emergence of postural strategies, it is essential to distinguish between results that can be explained by biomechanical reasons strictly and those reflecting the maturation of the central nervous system (CNS). To address this problem, we have studied our young subjects in situations requiring various types of adaptation. The studies dealing with adaptation of postural strategies aimed at testing short and long-term adaptation capacity of the CNS during imposed transient external biomechanical constraints in healthy children, and during chronic internal constraints in children with skeletal pathologies. In addition to maintenance of balance, another function of posture is to ensure the orientation of a body segment. It appears that the control of orientation and the control of balance both require the trunk as an initial reference frame involving a development from egocentric to exocentric postural control. It is concluded that the first step for children consists in building a repertoire of postural strategies, and the second step consists in learning to select the most appropriate postural strategy, depending on the ability to anticipate the consequence of the movement in order to maintain balance control and the efficiency of the task.


Effects of napping on cognitive function modulation in elderly adults with a morning chronotype: A nationwide survey.
Circadian rhythms and napping habits have been associated with cognitive function; however, little is known about the interaction effects on cognitive function. The present study examined the moderating effects of napping habits on the relationship between chronotypes and cognitive function decline in elderly adults. This cross-sectional study analysed data from the 2009 Taiwan National Health Interview Survey. A total of 1,724 elderly adults (aged ≥65 years) were included. Cognitive function was determined by the Mini-Mental State Examination. Participants' chronotypes were assessed using the midpoint of sleep. Napping habits were assessed by self-report questions. Demographic data, health-related factors and sleep characteristics of the participants were also assessed as potential confounding factors. A hierarchical regression will be used to test the moderating role of napping habits in the relationship between chronotype and cognitive function level after adjustment for potential confounders. A significant interaction effect between chronotype and napping habits was observed with the MMSE score (b = -0.29, SE = 0.14, p = 0.03). Morning-type elderly patients with nap durations >60 min exhibited significantly higher MMSE scores than non-nappers (F = 7.48; p = 0.00). This nationwide survey confirms that in morning-type elderly adults, nap durations >60 min are associated with less cognitive decline; however, napping habits are not associated with cognitive function level in evening-type and intermediate-type elderly adults.


Mithramycin downregulates proinflammatory cytokine-induced matrix metalloproteinase gene expression in articular chondrocytes.
Interleukin-1 (IL-1), IL-17 and tumor necrosis factor alpha (TNF-alpha) are the main proinflammatory cytokines implicated in cartilage breakdown by matrix metalloproteinase (MMPs) in arthritic joints. We studied the impact of an anti-neoplastic antibiotic, mithramycin, on the induction of MMPs in chondrocytes. MMP-3 and MMP-13 gene expression induced by IL-1beta, TNF-alpha and IL-17 was downregulated by mithramycin in human chondrosarcoma SW1353 cells and in primary human and bovine femoral head chondrocytes. Constitutive and IL-1-stimulated MMP-13 levels in bovine and human cartilage explants were also suppressed. Mithramycin did not significantly affect the phosphorylation of the mitogen-activated protein kinases, extracellular signal-regulated kinase, p38 and c-Jun N-terminal kinase. Despite effective inhibition of MMP expression by mithramycin and its potential to reduce cartilage degeneration, the agent might work through multiple unidentified mechanisms.


Hepcidin, a key regulator of iron metabolism, is transcriptionally activated by p53.
Hepcidin is an iron-regulatory protein that is upregulated in response to increased iron or inflammatory stimuli. Hepcidin reduces serum iron and induces iron sequestration in the reticuloendothelial macrophages - the hallmark of anaemia of inflammation. Iron deprivation is used as a defense mechanism against infection, and it also has a beneficial effect on the control of cancer. The tumour-suppressor p53 transcriptionally regulates genes involved in growth arrest, apoptosis and DNA repair, and perturbation of p53 pathways is a hallmark of the majority of human cancers. This study inspected a role of p53 in the transcriptional regulation of hepcidin. Based on preliminary bioinformatics analysis, we identified a putative p53 response-element (p53RE) contained in the hepcidin gene (HAMP) promoter. Chromatin immunoprecipitation (ChIP), reporter assays and a temperature sensitive p53 cell-line system were used to demonstrate p53 binding and activation of the hepcidin promoter. p53 bound to hepcidin p53RE in vivo, andthis p53RE could confer p53-dependent transcriptional activation. Activation of p53 increased hepcidin expression, while silencing of p53 resulted in decreased hepcidin expression in human hepatoma cells. Taken together, these results define HAMP as a novel transcriptional target of p53. We hypothesise that hepcidin upregulation by p53 is part of a defence mechanism against cancer, through iron deprivation. Hepcidin induction by p53 might be involved in the pathogenesis of anaemia accompanying cancer.


Decisions on multiple sclerosis immunotherapy: new treatment complexities urge patient engagement.
For patients with multiple sclerosis (MS) involvement in treatment decisions becomes ever more imperative. Recently new therapeutic options have become available for the treatment of MS and more will be licensed in the near future. Although more efficacious and easier to administer, the new drugs pose increased risks of severe side effects. Also, new diagnostic criteria lead to more and earlier MS diagnoses. Facing increasingly complex decisions, patients need up-to-date evidence-based information and decision support systems in order to make informed decision together with physicians based on their autonomy preferences. This article summarizes recently terminated and ongoing trials on MS patient education and decision aids conducted by the authors' study groups. Programs on relapse management, immunotherapy, and for patients with suspected and early MS have been developed and evaluated in randomized controlled clinical trials. It could be shown that the programs successfully increase knowledge and allow patients to make informed decisions based on their preferences. For the near future, we aim to develop a modular program for all relevant decisions in MS to increase patients' self-management and empower patients to develop their individual approach with the disease. Faced by a disease with many uncertainties, this should enhance patients' sense of control. Still, it remains a challenge to adequately assess decision quality. Therefore, a study in six European and one Australian centers will start soon aiming to establish adequate tools to assess decision-making quality.


Ontogenetic changes in cutaneous and branchial ionocytes and morphology in yellowfin tuna (Thunnus albacares) larvae.
The development of osmoregulatory and gas exchange organs was studied in larval yellowfin tuna (Thunnus albacares) from 2 to 25 days post-hatching (2.9-24.5 mm standard length, SL). Cutaneous and branchial ionocytes were identified using Na+/K+-ATPase immunostaining and scanning electron microscopy. Cutaneous ionocyte abundance significantly increased with SL, but a reduction in ionocyte size and density resulted in a significant decrease in relative ionocyte area. Cutaneous ionocytes in preflexion larvae had a wide apical opening with extended microvilli; however, microvilli retracted into an apical pit from flexion onward. Lamellae in the gill and pseudobranch were first detected ~ 3.3 mm SL. Ionocytes were always present on the gill arch, first appeared in the filaments and lamellae of the pseudobranch at 3.4 mm SL, and later in gill filaments at 4.2 mm SL, but were never observed in the gill lamellae. Unlike the cutaneous ionocytes, gill and pseudobranch ionocytes had a wide apical opening with extended microvilli throughout larval development. The interlamellar fusion, a specialized gill structure binding the lamellae of ram-ventilating fish, began forming by ~ 24.5 mm SL and contained ionocytes, a localization never before reported. Ionocytes were retained on the lamellar fusions and also found on the filament fusions of larger sub-adult yellowfin tuna; however, sub-adult gill ionocytes had apical pits. These results indicate a shift in gas exchange and NaCl secretion from the skin to branchial organs around the flexion stage, and reveal novel aspects of ionocyte localization and morphology in ram-ventilating fishes.


Paclitaxel-eluting stents show superiority to balloon angioplasty and bare metal stents in femoropopliteal disease: twelve-month Zilver PTX randomized study results.
Sustained benefits of drug-eluting stents in femoropopliteal arteries have not been demonstrated. This prospective, multinational, randomized study was designed to compare the 12-month safety and effectiveness of a polymer-free, paclitaxel-coated nitinol drug-eluting stent (DES) with percutaneous transluminal angioplasty (PTA) and provisional bare metal stent (BMS) placement in patients with femoropopliteal peripheral artery disease. Patients were randomly assigned to primary DES implantation (n=236) or PTA (n=238). Demographics and lesion characteristics were similar between groups (eg, average lesion length, approximately 65±40 mm). One hundred twenty patients had acute PTA failure and underwent secondary random assignment to provisional DES (n=61) or BMS (n=59). Primary end points were the 12-month rates of event-free survival and patency in the primary DES and PTA groups. Compared with the PTA group, the primary DES group exhibited superior 12-month event-free survival (90.4% versus 82.6%; P=0.004) and primary patency (83.1% versus 32.8%; P<0.001), satisfying the primary hypotheses. In the secondary evaluations, (1) the primary DES group exhibited superior clinical benefit compared with the PTA group (88.3% versus 75.8%; P<0.001), (2) the provisional DES group exhibited superior primary patency (89.9% versus 73.0%; P=0.01) and superior clinical benefit (90.5% and 72.3%, P=0.009) compared with the provisional BMS group, and (3) the stent fracture rate (both DES and BMS) was 0.9% (4/457). Femoropopliteal peripheral artery disease treatment with the paclitaxel-eluting stent was associated with superior 12-month outcomes compared with PTA and provisional BMS placement.


Cerebral mitochondrial electron transport chain dysfunction in multiple system atrophy and Parkinson's disease.
Multiple system atrophy (MSA) is a neurodegenerative disease characterised by glial cytoplasmic inclusions (GCIs), containing α-synuclein. Mutated COQ2, encoding an enzyme essential for co-enzyme Q10 (CoQ10) biosynthesis, has been associated with MSA. CoQ10 is an electron carrier in the mitochondrial electron transport chain (ETC) and antioxidant. It has been shown to be deficient in MSA brain tissue, thus implicating mitochondrial dysfunction in MSA. To investigate mitochondrial dysfunction in MSA further we examined ETC activity in MSA and control brain tissue, compared with Parkinson's disease (PD) where mitochondrial dysfunction is known to be important. Using cerebellar and occipital white matter ETC complex I, II/III and IV activities were measured spectrophotometrically, selected individual components of the ETC were assessed by immunoblotting and cellular complex IV activity was analysed by enzyme histochemistry. We show decreased complex II/III activity with increased complex I and IV activity in MSA cerebellar white matter. This corresponds with the deficit in CoQ10 previously described in MSA and reflects the high regional pathological burden of GCIs. This study highlights mitochondrial dysfunction in MSA pathogenesis, suggests an influence on selective regional vulnerability to disease and points to shared disease mechanisms in α-synucleinopathies.


Relationship between ethnicity and glycemic control, lipid profiles, and blood pressure during the first 9 years of type 2 diabetes: U.K. Prospective Diabetes Study (UKPDS 55).
To assess the relationship among self-reported ethnicity, metabolic control, and blood pressure during treatment of type 2 diabetes. We studied 2,999 newly diagnosed type 2 diabetic patients recruited to the U.K. Prospective Diabetes Study who were randomized to conventional or intensive glucose control policies if their fasting plasma glucose levels remained >6 mmol/l after a dietary run-in. A total of 2,484 patients (83%) were white Caucasian (WC), 265 patients (9%) were Afro-Caribbean (AC), and 250 patients (8%) were Asian of Indian origin (IA). Variables were assessed at 3, 6, and 9 years. During the 9-year study period, body weight increased more in WC patients (mean 5.0 kg) than in AC (3.0 kg) and IA (2.5 kg) patients (P < 0.001). After adjusting for age, sex, baseline value, treatment allocation, and change in weight, there were no consistent ethnic differences in mean change in fasting plasma glucose or HbA(1c). After adjustment for antihypertensive therapy, increase in systolic blood pressure at 9 years was greatest in AC patients (7 mmHg; P < 0.01 vs. WC patients). Mean diastolic blood pressure, total cholesterol, and LDL cholesterol decreased progressively during the 9 years in each group. In AC patients, the mean increase in HDL cholesterol (0.16 mmol/l) at 3 years, maintained to 9 years, and the mean decrease in plasma triglyceride level (0.4 mmol/l) at 9 years were greater than in WC and IA patients (P < 0.001). This study shows important ethnic differences in body weight, lipid profiles, and blood pressure, but not glycemic control, during 9 years after diagnosis of type 2 diabetes. AC patients maintained the most favorable lipid profiles, but hypertension developed in more AC patients than WC or IA patients. Ethnicity-specific glycemic control of type 2 diabetes seems unnecessary, but other risk factors need to be addressed independently.


Cruciferous Vegetables, Isothiocyanates, and Bladder Cancer Prevention.
Bladder cancer is a significant health burden due to its high prevalence, risk of mortality, morbidity, and high cost of medical care. Epidemiologic evidence suggests that diets rich in cruciferous vegetables, particularly broccoli, are associated with lower bladder cancer risk. Phytochemicals in cruciferous vegetables, such as glucosinolates, which are enzymatically hydrolyzed to bioactive isothiocyanates, are possible mediators of an anticancer effect. In vitro studies have shown inhibition of bladder cancer cell lines, cell cycle arrest, and induction of apoptosis by these isothiocyanates, in particular sulforaphane and erucin. Although not yet completely understood, many mechanisms of anticancer activity at the steps of cancer initiation, promotion, and progression have been attributed to these isothiocyanates. They target multiple pathways including the adaptive stress response, phase I/II enzyme modulation, pro-growth, pro-survival, pro-inflammatory signaling, angiogenesis, and even epigenetic modulation. Multiple in vivo studies have shown the bioavailability of isothiocyanates and their antitumoral effects. Although human studies are limited, they support oral bioavailability with reasonable plasma and urine concentrations achieved. Overall, both cell and animal studies support a potential role for isothiocyanates in bladder cancer prevention and treatment. Future studies are necessary to examine clinically relevant outcomes and define guidelines on ameliorating the bladder cancer burden.


Nanostructured diatom earth SiO2 negative electrodes with superior electrochemical performance for lithium ion batteries.
Diatomaceous earth (DE) is a naturally occurring silica source constituted by fossilized remains of diatoms, a type of hard-shelled algae, which exhibits a complex hierarchically nanostructured porous silica network. In this work, we analyze the positive effects of reducing DE SiO2 particles to the sub-micrometer level and implementing an optimized carbon coating treatment to obtain DE SiO2 anodes with superior electrochemical performance for Li-ion batteries. Pristine DE with an average particle size of 17 μm is able to deliver a specific capacity of 575 mA h g-1 after 100 cycles at a constant current of 100 mA g-1, and reducing the particle size to 470 nm enhanced the reversible specific capacity to 740 mA h g-1. Ball-milled DE particles were later subjected to a carbon coating treatment involving the thermal decomposition of a carbohydrate precursor at the surface of the particles. Coated ball-milled silica particles reached stable specific capacities of 840 mA h g-1 after 100 cycles and displayed significantly improved rate capability, with discharge specific capacities increasing from 220 mA h g-1 (uncoated ball-milled SiO2) to 450 mA h g-1 (carbon coated ball-milled SiO2) at 2 A g-1. In order to trigger SiO2 reactivity towards lithium, all samples were subjected to an electrochemical activation procedure prior to electrochemical testing. XRD measurements on the activated electrodes revealed that the initial crystalline silica was completely converted to amorphous phases with short range ordering, therefore evidencing the effective role of the activation procedure.


Molecular and morphological description of a Hepatozoon species in reptiles and their ticks in the Northern Territory, Australia.
Ticks, representing 3 species of Amblyomma, were collected from the water python (Liasis fuscus) and 3 additional reptile species in the Northern Territory, Australia, and tested for the presence of Hepatozoon sp., the most common blood parasites of snakes. In addition, blood smears were collected from 5 reptiles, including the water python, and examined for the presence of the parasite. Hepatozoon sp. DNA was detected in all tick and reptile species, with 57.7% of tick samples (n = 187) and 35.6% of blood smears (n=35) showing evidence of infection. Phylogenetic analysis of the 18S rRNA gene demonstrated that half of the sequences obtained from positive tick samples matched closest with a Hepatozoon species previously identified in the water python population. The remaining sequences were found to be more closely related to mammalian and amphibian Hepatozoon species. This study confirms that species of Amblyomma harbor DNA of the same Hepatozoon species detected in the water pythons. The detection of an additional genotype suggests the ticks may be exposed to 2 Hepatozoon species, providing further opportunity to study multiple host-vector-parasite relationships.


Three missense variants of metabolic syndrome-related genes are associated with alpha-1 antitrypsin levels.
Alpha-1 antitrypsin (AAT) encoded by SERPINA1 is an acute-phase inflammation marker, and AAT deficiency (AATD) is known as one of the common genetic disorders in European populations. However, no genetic determinants to AAT levels apart from the SERPINA gene clusters have been identified to date. Here we perform a genome-wide association study of serum AAT levels followed by a two-staged replication study recruiting a total of 9,359 Japanese community-dwelling population. Three missense variants of metabolic syndrome-related genes, namely, rs671 in ALDH2, rs1169288 in HNF1A and rs1260326 in GCKR, significantly associate with AAT levels (P≤1.5 × 10(-12)). Previous reports have shown the functional relevance of ALDH2 and HNF1A to AAT. We observe a significant interaction of rs671 and alcohol consumption on AAT levels. We confirm the association between AAT and rs2896268 in SERPINA1, which is independent of known causative variants of AATD. These findings would support various AAT functions including metabolic processes.


The effect of an insect herbivore and a rust fungus individually, and combined in sequence, on the growth of two Rumex species.
The chrysomelid beetle Gastrophysa viridula and the rust fungus Urmnyces rumicis both occur on leaves of Rumex crispus and R. obtusifolius. We investigated the effect of beetle grazing or rust infection individually and when combined in sequence on the growth of their hosts in the field. Singly, beetle or rust reduced leaf area and plant biomass; the effect was greater on R. crispus, and rust caused greater damage than the beetle. Beetle grazing with subsequent rust infection caused damage no greater than that caused by rust alone, although on R. obtusifolius damage was greater than that from beetle grazing alone. Rust infection of R. obtusifolius with subsequent beetle grazing produced damage similar to that from other treatments; involving rust infection. In R. crispus this treatment produced the greatest reduction in biomass, The reductions in root and total plant weight from rust infection with subsequent beetle grazing were accurately predicted by a model including the damage produced by beetle and rust alone and the length of time each was present on the plant. This model also predicted accurately the damage to R. obtusifolius from the beetle followed by rust treatment, but over-estimated by up to 40% the damage to R. crispus. This can be explained mainly by an inhibition of rust infection by beetle grazing.


Improving text recall with multiple summaries.
QuikScan (QS) is an innovative design that aims to improve accessibility, comprehensibility, and subsequent recall of expository text by means of frequent within-document summaries that are formatted as numbered list items. The numbers in the QS summaries correspond to numbers placed in the body of the document where the summarized ideas are discussed in full. To examine the influence of QS summaries on participants' perceptions of text quality (i.e., comprehensibility, structure, and interest) and recall, an experimental - control group design compared the effects of a QS text with a structured abstract (SA) text. Forty psychology students participated voluntarily or received course credits. Students first read a control (SA) or experimental (QS) text on flashbulb memory (FBM). Next, their perceptions of text quality were measured through a questionnaire. Recall was assessed with an open answer test with items for facts, comprehension and higher order information. Perceptions of text quality did not vary across conditions. But QS did lead to significantly and substantially (d= 1.57) higher overall recall scores. Participants with the QS text performed significantly better on all item types than participants with the SA text. Studying a QS text led to a substantial improvement in recall compared to an SA text. Further research is needed to examine how readers study QS texts and whether a text model hypothesis or a repetition effect hypothesis accounts for the effectiveness. The first hypothesis posits that the QS summaries support the reader in constructing a text schema. The second attributes the effects of these summaries to their repetition of text topics.


Suicidal drowning deaths in northern Sweden 1992-2009 - The role of mental disorder and intoxication.
Suicides by drowning have received limited attention by researchers. A recent finding that almost one-third of all drowning deaths in Sweden were classified as suicide instigated this study. We identified 129 cases of suicide by drowning in Northern Sweden and analyzed the circumstances and the psychiatric history prior to the suicide. Information was obtained from autopsy, police and medical records, as well as from the National Inpatient Register. One-third of the suicide victims had previously attempted suicide and half of the victims had been hospitalized due to mental health problems. One-third of these had left the hospital less than one week before the suicide. Alcohol and psychoactive drugs were present in 16% and 62% of the cases, respectively. A history of mental disorder and previous suicide attempt (s), especially by drowning, is an ominous combination necessitating efficient clinical identification, treatment and follow-up if a complete suicide is to be prevented.


Childhood motor skill proficiency as a predictor of adolescent physical activity.
Cross-sectional evidence has demonstrated the importance of motor skill proficiency to physical activity participation, but it is unknown whether skill proficiency predicts subsequent physical activity. In 2000, children's proficiency in object control (kick, catch, throw) and locomotor (hop, side gallop, vertical jump) skills were assessed in a school intervention. In 2006/07, the physical activity of former participants was assessed using the Australian Physical Activity Recall Questionnaire. Linear regressions examined relationships between the reported time adolescents spent participating in moderate-to-vigorous or organized physical activity and their childhood skill proficiency, controlling for gender and school grade. A logistic regression examined the probability of participating in vigorous activity. Of 481 original participants located, 297 (62%) consented and 276 (57%) were surveyed. All were in secondary school with females comprising 52% (144). Adolescent time in moderate-to-vigorous and organized activity was positively associated with childhood object control proficiency. Respective models accounted for 12.7% (p = .001), and 18.2% of the variation (p = .003). Object control proficient children became adolescents with a 10% to 20% higher chance of vigorous activity participation. Object control proficient children were more likely to become active adolescents. Motor skill development should be a key strategy in childhood interventions aiming to promote long-term physical activity.


Phytoestrogen Suppresses Efflux of the Diagnostic Marker Protoporphyrin IX in Lung Carcinoma.
One promising method to visualize cancer cells is based on the detection of the fluorescent photosensitizer protoporphyrin IX (PpIX) synthesized from 5-aminolevulinic acid (ALA), but this method cannot be used in cancers that exhibit poor PpIX accumulation. PpIX appears to be pumped out of cancer cells by the ABC transporter G2 (ABCG2), which is associated with multidrug resistance. Genistein is a phytoestrogen that appears to competitively inhibit ABCG2 activity. Therefore, we investigated whether genistein can promote PpIX accumulation in human lung carcinoma cells. Here we report that treatment of A549 lung carcinoma cells with genistein or a specific ABCG2 inhibitor promoted ALA-mediated accumulation of PpIX by approximately 2-fold. ABCG2 depletion and overexpression studies further revealed that genistein promoted PpIX accumulation via functional repression of ABCG2. After an extended period of genistein treatment, a significant increase in PpIX accumulation was observed in A549 cells (3.7-fold) and in other cell lines. Systemic preconditioning with genistein in a mouse xenograft model of lung carcinoma resulted in a 1.8-fold increase in accumulated PpIX. Long-term genistein treatment stimulated the expression of genes encoding enzymes involved in PpIX synthesis, such as porphobilinogen deaminase, uroporphyrinogen decarboxylase, and protoporphyrinogen oxidase. Accordingly, the rate of PpIX synthesis was also accelerated by genistein pretreatment. Thus, our results suggest that genistein treatment effectively enhances ALA-induced PpIX accumulation by preventing the ABCG2-mediated efflux of PpIX from lung cancer cells and may represent a promising strategy to improve ALA-based diagnostic approaches in a broader set of malignancies. Cancer Res; 76(7); 1837-46. ©2016 AACR.


The microenvironment in hepatocyte regeneration and function in rats with advanced cirrhosis.
In advanced cirrhosis, impaired function is caused by intrinsic damage to the native liver cells and from the abnormal microenvironment in which the cells reside. The extent to which each plays a role in liver failure and regeneration is unknown. To examine this issue, hepatocytes from cirrhotic and age-matched control rats were isolated, characterized, and transplanted into the livers of noncirrhotic hosts whose livers permit extensive repopulation with donor cells. Primary hepatocytes derived from livers with advanced cirrhosis and compensated function maintained metabolic activity and the ability to secrete liver-specific proteins, whereas hepatocytes derived from cirrhotic livers with decompensated function failed to maintain metabolic or secretory activity. Telomere studies and transcriptomic analysis of hepatocytes recovered from progressively worsening cirrhotic livers suggest that hepatocytes from irreversibly failing livers show signs of replicative senescence and express genes that simultaneously drive both proliferation and apoptosis, with a later effect on metabolism, all under the control of a central cluster of regulatory genes, including nuclear factor κB and hepatocyte nuclear factor 4α. Cells from cirrhotic and control livers engrafted equally well, but those from animals with cirrhosis and failing livers showed little initial evidence of proliferative capacity or function. Both, however, recovered more than 2 months after transplantation, indicating that either mature hepatocytes or a subpopulation of adult stem cells are capable of full recovery in severe cirrhosis. Transplantation studies indicate that the state of the host microenvironment is critical to the regenerative potential of hepatocytes, and that a change in the extracellular matrix can lead to regeneration and restoration of function by cells derived from livers with end-stage organ failure.


Biventricular Rupture with Extracardiac Left-to-Right Shunt Complicating Acute Myocardial Infarction.
Simultaneous rupture of the left and right ventricles is an extremely rare mechanical complication of acute myocardial infarction (MI). When associated with the formation of a false aneurysm, an extracardiac left-to-right shunt may occur. We summarized all published data describing this unique condition. We searched the PubMed and Google Scholar databases for case reports in peer-reviewed journals from 1 January 1980 to 1 May 2015. We identified 16 articles describing 17 cases. In all but 1 case, biventricular wall rupture (BVWR) resulted from an inferior MI. The clinical presentations of BVWR were variable and included cardiogenic shock, congestive heart failure and an absence of any cardiac symptoms. In most cases, there was a hemodynamically significant left-to-right shunt, with pulmonary to systemic blood flow (Qp/Qs) >2. Diagnostic difficulties were reported in most cases, and some patients were initially misdiagnosed as having ventricular septal rupture (VSR). Surgical closure of the defect was successful in most cases, and some asymptomatic patients were managed conservatively. BVWR with an intact interventricular septum and extracardiac left-to-right shunt is a rare mechanical complication of acute MI, often misdiagnosed as VSR. It has a variable clinical course, probably related to the magnitude of the shunt.


Motoric cognitive risk syndrome and the risk of dementia.
Despite growing evidence of links between gait and cognition in aging, cognitive risk assessments that incorporate motoric signs have not been examined. We sought to validate a new Motoric Cognitive Risk (MCR) syndrome to identify individuals at high risk of developing dementia. We evaluated 997 community residing individuals aged 70 and older participating in the Einstein Aging Study over a median follow-up time of 36.9 months. MCR syndrome was defined as presence of cognitive complaints and slow gait (one standard deviation below age- and sex-specific gait speed means) in nondemented individuals. Cox models were used to evaluate the effect of MCR syndrome on the risk of developing dementia and subtypes. Fifty-two participants met criteria for MCR syndrome at baseline with a prevalence of 7% (95% CI: 5-9%). Prevalence of MCR increased with age. Participants with MCR were at higher risk of developing dementia (hazard ratio [HR] adjusted for age, sex, and education: 3.27, 95% CI: 1.55-6.90) and vascular dementia (adjusted HR: 12.81, 95% CI: 4.98-32.97). The association of MCR with risk of dementia or vascular dementia remained significant even after accounting for other confounders and diagnostic overlap with "cognitive" mild cognitive impairment syndrome subtypes. A motor-based MCR syndrome provides a clinical approach to identify individuals at high risk for dementia, especially vascular dementia, to target for further investigations and who may benefit from preventive interventions.


High precision alignment procedure for an ellipsometer.
Beam deviation in the polarizing elements is identified as a significant source of error in existing ellipsometer alignment procedures. A high precision alignment procedure that eliminates these errors is described. This procedure is less time consuming than previous methods, and its accuracy is comparable to the limit of resolution of the ellipsometer (typically 0.01-0.005 degrees ). A further advantage of this procedure is that it provides a precise method for the alignment of specimens and lasers with the ellipsometer.


In Vivo Efficacy of Umbilical Cord Blood Stem Cell-Derived NK Cells in the Treatment of Metastatic Colorectal Cancer.
Therapeutic monoclonal antibodies against the epidermal growth factor receptor (EGFR) act by inhibiting EGFR downstream signaling and by eliciting a natural killer (NK) cell-mediated antitumor response. The IgG1 mAb cetuximab has been used for treatment of RASwt metastatic colorectal cancer (mCRC) patients, showing limited efficacy. In the present study, we address the potential of adoptive NK cell therapy to overcome these limitations investigating two allogeneic NK cell products, i.e., allogeneic activated peripheral blood NK cells (A-PBNK) and umbilical cord blood stem cell-derived NK cells (UCB-NK). While cetuximab monotherapy was not effective against EGFR- RASwt, EGFR+ RASmut, and EGFR+ BRAFmut cells, A-PBNK were able to initiate lysis of EGFR+ colon cancer cells irrespective of RAS or BRAF status. Cytotoxic effects of A-PBNK (but not UCB-NK) were further potentiated significantly by coating EGFR+ colon cancer cells with cetuximab. Of note, a significantly higher cytotoxicity was induced by UCB-NK in EGFR-RASwt (42 ± 8 versus 67 ± 7%), EGFR+ RASmut (20 ± 2 versus 37 ± 6%), and EGFR+ BRAFmut (23 ± 3 versus 43 ± 7%) colon cancer cells compared to A-PBNK and equaled the cytotoxic efficacy of the combination of A-PBNK and cetuximab. The antitumor efficacy of UCB-NK cells against cetuximab-resistant human EGFR+ RASmut colon cancer cells was further confirmed in an in vivo preclinical mouse model where UCB-NK showed enhanced antitumor cytotoxicity against colon cancer independent of EGFR and RAS status. As UCB-NK have been proven safe in a recently conducted phase I clinical trial in acute myeloid leukemia, a fast translation into clinical proof of concept for mCRC could be considered.


Endovascular therapy of wake-up strokes in the modern era of stent retriever thrombectomy.
Endovascular treatment of wake-up strokes (WUS) has been previously described, mostly with the use of pharmacological thrombolysis or first generation thrombectomy devices. To describe outcomes of WUS treated with modern endovascular therapy since the Food and Drug Administration approval of stent retrievers, and to identify predictors of good clinical outcome in this population of stroke patients. We performed a multicenter retrospective analysis of consecutive patients with WUS who underwent thrombectomy with stent retrievers Trevo (Stryker, Kalamazoo, Michigan, USA) and Solitaire FR (Covidien, Irvine, California, USA), or primary aspiration thrombectomy. We correlated favorable clinical outcomes with demographic, clinical, and technical characteristics. 52 patients were included in this study; 46 (88%) cases were treated with stent retrievers and 6 (12%) were treated with primary aspiration thrombectomy alone. Successful recanalization (Thrombolysis in Cerebral Infarction (TICI) 2b/3) was achieved in 36 (69%) patients. Favorable clinical outcome at 3 months, defined as a modified Rankin Scale score of 0-2, was achieved in 25 (48%) patients. Duration of intervention <30 min and its success, defined as TICI 2b/3 recanalization, were strong predictors of favorable clinical outcome at 90 days (p<0.001 and p<0.0001, respectively). Our study indicates that endovascular treatment of WUS with stent retrievers and aspiration thrombectomy is safe and effective.


Does airway surgery lower serum lipid levels in obstructive sleep apnea patients? A retrospective case review.
Obstructive sleep apnea (OSA) is tightly linked to increased cardiovascular disease. Surgery is an important method to treat OSA, but its effect on serum lipid levels in OSA patients is unknown. We aimed to evaluate the effect of upper airway surgery on lipid profiles. We performed a retrospective review of 113 adult patients with OSA who underwent surgery (nasal or uvulopalatopharyngoplasty [UPPP]) at a major, urban, academic hospital in Beijing from 2012 to 2013 who had preoperative and postoperative serum lipid profiles. Serum TC (4.86±0.74 to 4.69±0.71) and LP(a) (median 18.50 to 10.90) all decreased significantly post-operatively (P<0.01, 0.01, respectively), with no changes in serum HDL, LDL, or TG (P>0.05, all). For UPPP patients (n=51), serum TC, HDL and LP(a) improved (P=0.01, 0.01,<0.01, respectively). For nasal patients (n=62), only the serum LP(a) decreased (P<0.01). In patients with normal serum lipids at baseline, only serum LP(a) decreased (P<0.01). In contrast, in patients with isolated hypertriglyceridemia, the serum HDL, TG and LP(a) showed significant improvements (P=0.02, 0.03, <0.01, respectively). In patients with isolated hypercholesterolemia, the serum LP(a) decreased significantly (P=0.01), with a similar trend for serum TC (P=0.06). In patients with mixed hyperlipidemia, the serum TC and LDL also decreased (P=0.02, 0.03, respectively). Surgery may improve blood lipid levels in patients with OSA, especially in patients with preoperative dyslipidemia, potentially yielding a major benefit in metabolism and cardiovascular sequelae. Prospective studies should examine this potential metabolic effect of airway surgery for OSA.


On Error-Related Potentials During Sensorimotor-Based Brain-Computer Interface: Explorations With a Pseudo-Online Brain-Controlled Speller.
Objective: Brain-computer interface (BCI) spelling is a promising communication solution for people in paralysis. Currently, BCIs suffer from imperfect decoding accuracy which calls for methods to handle spelling mistakes. Detecting error-related potentials (ErrPs) has been early identified as a potential remedy. Nevertheless, few works have studied the elicitation of ErrPs during engagement with other BCI tasks, especially when BCI feedback is provided continuously. Methods: Here, we test the possibility of correcting errors during pseudo-online Motor Imagery (MI) BCI spelling through ErrPs, and investigate whether BCI feedback hinders their generation. Ten subjects performed a series of MI spelling tasks with and without observing BCI feedback. Results: The average pseudo-online ErrP detection accuracy was found to be significantly above the chance level in both conditions and did not significantly differ between the two (74% with, and 78% without feedback). Conclusions: Our results support the possibility to detect ErrPs during MI-BCI spelling and suggest the absence of any BCI feedback-related interference.


Single-Trial EEG-EMG coherence analysis reveals muscle fatigue-related progressive alterations in corticomuscular coupling.
Voluntary muscle fatigue is a progressive process. A recent study demonstrated muscle fatigue-induced weakening of functional corticomuscular coupling measured by coherence between the brain [electroencephalogram (EEG)] and muscle [electromyogram (EMG)] signals after a relatively long-duration muscle contraction. Comparing the EEG-EMG coherence before versus after fatigue or between data of two long-duration time blocks is not adequate to reveal the dynamic nature of the fatigue process. The purpose of this study was to address this issue by quantifying single-trial EEG-EMG coherence and EEG, EMG power based on wavelet transform. Eight healthy subjects performed 200 maximal intermittent handgrip contractions in a single session with handgrip force, EEG and EMG signals acquired simultaneously. The EEG and EMG data during each 2-s handgrip was subjected to single trial EEG-EMG wavelet energy spectrum and coherence computation. The EEG-EMG coherence and energy spectrum at beta (15 ~ 35 Hz) and gamma (35-50 Hz) frequency bands were statistically analyzed in 2-block (75 trials per block), 5-block (30 trials/block), and 10-block (15 trials/block) data settings. The energy of both the EEG and EMG signals decreased significantly with muscle fatigue. The EEG-EMG coherence had a significant reduction for the 2-block comparison. More detailed dynamical changing and inter-subject variation of the EEG-EMG coherence and energy were revealed by 5- and 10-block comparisons. These results show feasibility of wavelet transform-based measurement of the EEG-EMG coherence and corresponding energy based on single-trial data, which provides extra information to demonstrate a time course of dynamic adaptations of the functional corticomuscular coupling, as well as brain and muscle signals during muscle fatigue.


The leader as a retention specialist.
Retention problems will plague us for the near future. Blanchard and Waghorn (1997) believe that everyone wants to be magnificent, and not just ordinary. Unfortunately, often we expect only the ordinary and not the magnificent. People respond to our expectations. We can help staff to achieve magnificence by providing unconditional love and remembering that people are magnificent. Sometimes their behavior is a problem and certain things block the expression of their greatness, but all people are magnificent. If we create "sanctuaries of caring" where nurses and others are given the opportunity to achieve their innate nobility and magnificence, people will flock to participate in the noble and magnificent experience of patient care.


Cope's rule and the evolution of body size in Pinnipedimorpha (Mammalia: Carnivora).
Cope's rule describes the evolutionary trend for animal lineages to increase in body size over time. In this study, we tested the validity of Cope's rule for a marine mammal clade, the Pinnipedimorpha, which includes the extinct Desmatophocidae, and extant Phocidae (earless seals), Otariidae (fur seals and sea lions), and Odobenidae (walruses). We tested for the presence of Cope's rule by compiling a large dataset of body size data for extant and fossil pinnipeds and then examined how body size evolved through time. We found that there was a positive relationship between geologic age and body size. However, this trend is the result of differences between early assemblages of small-bodied pinnipeds (Oligocene to early Miocene) and later assemblages (middle Miocene to Pliocene) for which species exhibited greater size diversity. No significant differences were found between the number of increases or decreases in body size within Pinnipedimorpha or within specific pinniped clades. This suggests that the pinniped body size increase was driven by passive diversification into vacant niche space, with the common ancestor of Pinnipedimorpha occurring near the minimum adult body size possible for a marine mammal. Based upon the above results, the evolutionary history of pinnipeds does not follow Cope's rule.


An Unusual Carbohydrate Conformation is Evident in Moraxella catarrhalis Oligosaccharides.
Oligosaccharide structures derived from the lipooligosaccharide of M. catarrhalis show that the highly branched glucose-rich inner core of the oligosaccharide has an altered conformation compared to the most truncated tetra-glucose-Kdo lgt1/4Δ oligosaccharide structure. Addition of one residue each to the (1-4) and (1-6) chains to give the lgt2Δ oligosaccharide is the minimum requirement for this conformational change to occur. Extensive molecular modeling and NMR investigations have shown that the (1-3), (1-4), and (1-6) glycosidic linkages from the central α-D-Glcp have significantly altered conformational preferences between the two structures. For the lgt1/4Δ oligosaccharide the (1-3) and (1-4) linkage populates predominantly the syn minimum on the conformational free energy map and for the (1-6) linkage conformational flexibility is observed, which is supported by 1H-NMR T1 measurements. For the lgt2Δ oligosaccharide the unusual "(1-4)anti-ψ(1-6)gg" conformation, which could be confirmed by long-range NOE signals, is a dominant conformation in which the oligosaccharide is very compact with the terminal α-D-GlcNAc residue folding back towards the center of the molecule leading to an extensive intra-molecular hydrophobic interaction between the terminal residues. Comparing effective H-H distances, which were calculated for conformational sub-ensembles, with the NOE distances revealed that typically multiple conformations could be present without significantly violating the measured NOE restraints. For lgt2Δ the presence of more than one conformation is supported by the NOE data.


Death from Nitrous Oxide.
Nitrous oxide is an inflammable gas that gives no smell or taste. It has a history of abuse as long as its clinical use, and deaths, although rare, have been reported. We describe two cases of accidental deaths related to voluntary inhalation of nitrous oxide, both found dead with a gas mask covering the face. In an attempt to find an explanation to why the victims did not react properly to oncoming hypoxia, we performed experiments where a test person was allowed to breath in a closed system, with or without nitrous oxide added. Vital signs and gas concentrations as well as subjective symptoms were recorded. The experiments indicated that the explanation to the fact that neither of the descendents had reacted to oncoming hypoxia and hypercapnia was due to the inhalation of nitrous oxide. This study raises the question whether nitrous oxide really should be easily, commercially available.


Introducing a quality improvement programme to primary healthcare teams.
To evaluate a programme in which quality improvement was facilitated, based on principles of total quality management, in primary healthcare teams, and to determine its feasibility, acceptability, effectiveness, and the duration of its effect. Primary healthcare teams in Leicestershire (n = 147) were invited to take part in the facilitated programme. The programme comprised seven team meetings, led by a researcher, plus up to two facilitated meetings of quality improvement subgroups, appointed by each team to consider specific quality issues. To assess the effect and feasibility of the programme on improving the quality of care provided, the individual quality improvement projects undertaken by the teams were documented and opportunities for improvement were noted at each session by the facilitator. The programme's acceptability was assessed with questionnaires issued in the final session to each participant. To assess the long term impact on teams, interviews with team members were conducted 3 years after the programme ended. 10 of the 27 teams that initially expressed interest in the programme agreed to take part, and six started the programme. Of these, five completed their quality improvement projects and used several different quality tools, and three completed all seven sessions of the programme. The programme was assessed as appropriate and acceptable by the participants. Three years later, the changes made during the programme were still in place in three of the six teams. Four teams had decided to undertake the local quality monitoring programme, resourced and supported by the Health Authority. The facilitated programme was feasible, acceptable, and effective for a few primary healthcare teams. The outcomes of the programme can be sustained. Research is needed on the characteristics of teams likely to be successful in the introduction and maintenance of quality improvement programmes.


Protocol for a systematic review of the association between chronic stress during the life course and telomere length.
The effects of stress on ill health have become evident in recent years. Under acute stress situations, a cascade of physiological events helps the body mount an appropriate adaptive response. However, under chronic stress situations, this physiological response may lead to wear and tear on the body that accelerates the decline in physiological functioning and increases the risk of chronic conditions. Recent evidence for social stress experienced during childhood suggests serious consequences many years later, even later life. Telomere length, a marker of cell aging, may provide a link between chronic social stress and age-associated physical and mental decline and risk of chronic conditions. This study examines whether chronic social stress is associated with telomere length throughout the life course. We will perform a systematic review of the literature on the relationship between chronic social stress, for example, due to violence, extreme poverty, or caregiving of people with disabling conditions (exposure), and telomere length (outcome) by searching electronic databases in MEDLINE (PubMed interface), EMBASE (OVID interface), Cochrane Central (OVID interface) and gray literature from their start date onwards. We will limit the search to studies performed on human populations. Two reviewers will conduct standardized screening, eligibility assessment, data abstraction, and scientific quality assessment. All study designs investigating the association between chronic social stress and telomere length in healthy or diseased adults and children will be eligible for inclusion in the review. We will extract individual demographic and socioeconomic characteristics, research setting, method of measuring telomere length, reported outcome, and determinants of interest. Studies will also be stratified by 1) age into 3 groups: childhood (0 to 18 years), adulthood (19 to 64 years) and late life (65+); 2) cell type; 3) study design; and 4) telomere length assessment method. Where feasible, study results will be combined through meta-analyses to obtain a pooled measure of associations. Results will be reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) Statement. This systematic review will provide knowledge on the existing evidence for chronic social stress and its association with telomere lengths throughout the life course.


Generation of atypical pulmonary inflammatory responses in BALB/c mice after immunization with the native attachment (G) glycoprotein of respiratory syncytial virus.
The feasibility of using the highly purified native attachment (G) protein in a subunit vaccine against respiratory syncytial virus (RSV) was examined in a murine model with or without the fusion (F) protein of RSV and the adjuvant QS-21. The studies established that QS-21 was more potent than AIOH as an adjuvant for both F and G glycoproteins. Augmented antigen-dependent killer cell activity and complement-assisted serum neutralizing and anti-F and G protein immunoglobulin G2a antibody titers were observed. Immunization with G/QS-21 generated immune responses that were characterized by low levels of antigen-dependent killer cell activity, elevated levels of interleukin-5 (IL-5) and percentages of eosinophils in the bronchoalveolar lavage fluids after challenge, and splenic immunocytes that secreted IL-5 but not gamma interferon (IFN-gamma) after in vitro stimulation with purified whole virus antigens. The pulmonary eosinophilia was similar to that induced by a facsimile of a formalin-inactivated vaccine used in previous clinical trials and was prevented by prior in vivo treatment with anti-IL-5 but not with control immunoglobulin G or anti-IFN-gamma neutralizing monoclonal antibodies. Thus the data implied that vaccination with G/QS-21 generated helper T-cell immune responses that were type 2 in nature. Alternatively, the data suggested that the helper T-cell immune responses elicited by F/QS-21 were more type 1 in character. Neither eosinophilia nor elevated levels of IL-5 were observed in the lungs of mice after challenge. Noteworthy levels of antigen-dependent killer cell activity was observed, and splenic immunocytes secreted copious quantities of IFN-gamma. Immunization with a combination vaccine composed of highly purified native F and G proteins plus QS-21 (F+G/QS-21) resulted in augmented complement-assisted serum neutralizing antibody titers compared with vaccination with either F/QS-21 or G/QS-21 alone. However, following vaccination with F+G/QS-21, the bronchoalveolar lavage fluids contained significant increases in IL-5 and percentages of eosinophils after challenge, the spleen cells appeared to secrete less IFN-gamma after in vitro stimulation, and there was no evidence of increased numbers of antigen-dependent killer cell precursors. Taken together, the data imply that native G protein influences the nature of the immune responses elicited by F/QS-21. The results therefore suggest that G, not F, protein has more potential to bias the host for atypical pulmonary inflammatory responses.


Alkylation of a dimolybdenum SO bridge, subsequent reactions, and characterization of the thioperoxide bridge.
The SO bridge of the complex, [Mo(2)(NTo)(2)(S(2)P(OEt)(2))(2)(mu-O(2)CMe)(mu-SBn)(mu-SO)], 1, displayed nucleophilicity at O, giving alkylation products [Mo(2)(NTo)(2)(S(2)P(OEt)(2))(2)(mu-O(2)CMe)(mu-SBn)(mu-SOR)](+), 4(+), which contained the thioperoxide bridge. These cations were then subject to nucleophilic attack by two pathways. Debenzylation of the bridge thiolate in 4(+) afforded neutral [Mo(2)(NTo)(2)(S(2)P(OEt)(2))(2)(mu-O(2)CMe)(mu-S)(mu-SOR)], 5; de-esterification of a dithiophosphate ligand in 4(+) gave [Mo(2)(NTo)(2)(S(2)P(O)(OEt))(S(2)P(OEt)(2))(mu-O(2)CMe)(mu-SBn)(mu-SO)], 6, which contained a monoester, dithiophosphate ligand. Complex 1 gave a slow and clean reaction in the crystalline state, further demonstrating its nucleophilicity by attacking a neighboring molecule in its lattice. X-ray crystallography confirmed the thioperoxide linkage and revealed structural similarities of the Mo(2)(mu-SOR) unit to sulfenate esters (RSOR) and related derivatives.


Studies of Trypanosoma cruzi clones in inbred mice. I. A comparison of the course of infection of C3H/HEN- mice with two clones isolated from a common source.
Two single-cell-isolate cloned stocks of the Sylvio-X10 strain, recovered from an acute human Trypanosoma cruzi infection, were used to infect C3H/HEN mice. The Sylvio-X10/4 clone produced a chronic infection in mice; clone Sylvio-X10/7 produced an acute lethal infection under identical experimental conditions. The course of infection of mice with the Sylvio-X10/7 clone was characterized by higher peripheral blood parasitemia and greater tissue involvement, an earlier appearance of specific anti-T. cruzi plasma IgG and shorter survival times than in mice infected with the Sylvio-X10/4 clone. The course of infection in mice with the Sylvio-X10 strain was intermediate between that of the two clones. This is the first demonstration of the pluripotential pathogenetic nature of a T. cruzi strain due to genetic heterogeneity of the population of parasites that constitute the strain. This experimental system is highly stable and reproducible. Consequently, the use of inbred mice and T. cruzi clones appears to provide an excellent model to study factors which influence the course of Chagas' disease.


Applying social marketing in health care: communicating evidence to change consumer behavior.
Social marketing uses commercial marketing strategies to change individual and organizational behavior and policies. It has been effective on a population level across a wide range of public health and health care domains. There is limited evidence of the effectiveness of social marketing in changing health care consumer behavior through its impact on patient-provider interaction or provider behavior. Social marketers need to identify translatable strategies (e.g., competition analysis, branding, and tailored messages) that can be applied to health care provider and consumer behavior. Three case studies from social marketing illustrate potential strategies to change provider and consumer behavior. Countermarketing is a rapidly growing social marketing strategy that has been effective in tobacco control and may be effective in countering pharmaceutical marketing using specific message strategies. Informed decision making is a useful strategy when there is medical uncertainty, such as in prostate cancer screening and treatment. Pharmaceutical industry marketing practices offer valuable lessons for developing competing messages to reach providers and consumers. Social marketing is an effective population-based behavior change strategy that can be applied in individual clinical settings and as a complement to reinforce messages communicated on a population level. There is a need for more research on message strategies that work in health care and population-level effectiveness studies.


Patient-reported outcome labeling claims and measurement approach for metastatic castration-resistant prostate cancer treatments in the United States and European Union.
Metastatic castration-resistant prostate cancer (mCRPC) and its treatment significantly affect health-related quality of life (HRQOL). Our objectives were to evaluate and compare patient-reported outcome (PRO) claims granted by the Food and Drug Administration (FDA) and European Medicines Agency (EMA) for 5 recently approved mCRPC treatments and to examine key characteristics, development, and measurement properties of the PRO measures supporting these claims against current regulatory standards. Five products approved for treatment of mCRPC by the FDA and the EMA (2010-2013) were examined: enzalutamide, abiraterone, sipuleucel-T, cabazitaxel, and radium Ra 223 dichloride. United States (US) drug approval packages and European Public Assessment Reports were reviewed. PRO claims in the US labels and European Summaries of Product Characteristics and supporting measures were identified. For PRO measures supporting claims, a targeted literature review was conducted to identify information on key characteristics and measurement properties; this information was compared against FDA PRO guidance criteria. Nine PRO "claims" were granted across 4 of 5 products reviewed. The EMA granted more claims (7 claims-4 for pain, 3 for HRQOL) than the FDA (2 claims, both for pain). The Brief Pain Inventory-Short Form (BPI-SF) worst pain item supported most pain claims and was the only measure supporting US claims. EMA pain claims were supported by BPI-SF worst pain (n = 2) and average pain (n = 1) items and the McGill Pain Questionnaire Present Pain Intensity component (n = 1). EMA HRQOL claims were supported by the Functional Assessment of Cancer Therapy-Prostate Module (n = 2) and the EuroQol 5 Dimensions with visual analogue scale (n = 1). Pain and prostate cancer-specific HRQOL measures supporting claims met US regulatory standards for construct validity, reliability, and responsiveness; these properties were strongest for the BPI-SF worst pain item. Only the BPI-SF worst pain item has documented content validity in mCRPC. PRO label claims were commonly granted across the mCRPC products reviewed. Among the measures reviewed, only the BPI-SF worst pain item supported US label claims. The BPI-SF worst pain item is recommended for pain assessment for the evaluation of new mCRPC treatments.


Current Perspectives on Circulating Tumor DNA, Precision Medicine, and Personalized Clinical Management of Cancer.
Circulating tumor DNA (ctDNA) has recently emerged as a minimally invasive "liquid biopsy" tool in precision medicine. ctDNA-genomic DNA fragments that are released into the bloodstream after the active secretion of microvesicles or tumor cell lysis reflects tumor evolution and the genomic alterations present in primary and/or metastatic tumors. Notably, ctDNA analysis might allow the stratification of patients, the monitoring of the therapeutic response, and the establishment of an opportunity for early intervention independent of detection by imaging modalities or clinical symptoms. As oncology moves towards precision medicine, the information in ctDNA provides a means for the individual management of the patient based on their tumor's genetic profile. This review presents current evidence on the potential role for ctDNA in helping to guide individualized clinical treatment decisions for patients with melanoma, castration-resistant prostate cancer, breast cancer, metastatic colorectal cancer, and non-small cell lung cancer.


Higher-order organisation of extremely amplified, potentially functional and massively methylated 5S rDNA in European pikes (Esox sp.).
Pikes represent an important genus (Esox) harbouring a pre-duplication karyotype (2n = 2x = 50) of economically important salmonid pseudopolyploids. Here, we have characterized the 5S ribosomal RNA genes (rDNA) in Esox lucius and its closely related E. cisalpinus using cytogenetic, molecular and genomic approaches. Intragenomic homogeneity and copy number estimation was carried out using Illumina reads. The higher-order structure of rDNA arrays was investigated by the analysis of long PacBio reads. Position of loci on chromosomes was determined by FISH. DNA methylation was analysed by methylation-sensitive restriction enzymes. The 5S rDNA loci occupy exclusively (peri)centromeric regions on 30-38 acrocentric chromosomes in both E. lucius and E. cisalpinus. The large number of loci is accompanied by extreme amplification of genes (>20,000 copies), which is to the best of our knowledge one of the highest copy number of rRNA genes in animals ever reported. Conserved secondary structures of predicted 5S rRNAs indicate that most of the amplified genes are potentially functional. Only few SNPs were found in genic regions indicating their high homogeneity while intergenic spacers were more heterogeneous and several families were identified. Analysis of 10-30 kb-long molecules sequenced by the PacBio technology (containing about 40% of total 5S rDNA) revealed that the vast majority (96%) of genes are organised in large several kilobase-long blocks. Dispersed genes or short tandems were less common (4%). The adjacent 5S blocks were directly linked, separated by intervening DNA and even inverted. The 5S units differing in the intergenic spacers formed both homogeneous and heterogeneous (mixed) blocks indicating variable degree of homogenisation between the loci. Both E. lucius and E. cisalpinus 5S rDNA was heavily methylated at CG dinucleotides. Extreme amplification of 5S rRNA genes in the Esox genome occurred in the absence of significant pseudogenisation suggesting its recent origin and/or intensive homogenisation processes. The dense methylation of units indicates that powerful epigenetic mechanisms have evolved in this group of fish to silence amplified genes. We discuss how the higher-order repeat structures impact on homogenisation of 5S rDNA in the genome.


Serial gadolinium enhanced magnetic resonance imaging in multiple sclerosis.
Serial gadolinium-DTPA (Gd-DTPA) enhanced magnetic resonance imaging (MRI) was performed in 9 patients with multiple sclerosis (MS). On the first scan enhancing lesions were seen in 7 patients, all of whom were in acute relapse. Most enhancing lesions were asymptomatic. On the second scan (3 to 5 weeks later), persisting enhancement was seen in only 12/54 lesions which enhanced on the first scan. No lesion showed persisting enhancement on the third scan (after 6 months). Enhancement occurred in all 12 new lesion areas seen on the unenhanced second scan and in 8 of 15 new lesions seen on the third scan. Enhancement was also seen in 4 older lesions which had been nonenhancing on earlier scans. Relaxation time measurements demonstrated a high water content in some nonenhancing lesions, which could be due to a subtle blood-brain barrier disturbance not detected with Gd-DTPA. The results show that blood-brain barrier impairment is a consistent finding in new MS lesions detected with MRI. Gd-DTPA is a useful marker of new and biologically active lesions and should prove of value in monitoring therapeutic trials in MS.


How Other Countries Use Deprivation Indices-And Why The United States Desperately Needs One.
Integrating public health and medicine to address social determinants of health is essential to achieving the Triple Aim of lower costs, improved care, and population health. There is intense interest in the United States in using social determinants of health to direct clinical and community health interventions, and to adjust quality measures and payments. The United Kingdom and New Zealand use data representing aspects of material and social deprivation from their censuses or from administrative data sets to construct indices designed to measure socioeconomic variation across communities, assess community needs, inform research, adjust clinical funding, allocate community resources, and determine policy impact. Indices provide these countries with comparable data and serve as a universal language and tool set to define organizing principles for population health. In this article we examine how these countries develop, validate, and operationalize their indices; explore their use in policy; and propose the development of a similar deprivation index for the United States.


The Effects of Exercise on Pain and Reproductive Performance in Female Pregnant Mice With Neuropathic Pain.
Pain can have negative, physiological and psychological impacts on pregnancy. Pregnant women are fearful of using pain medication because of teratogenic effects. In this study, we evaluated whether exercise could lower pain sensitivity in pregnant mice with neuropathic pain and reduce the negative effects of maternal pain on newborns. We randomly assigned 32 female mice to one of four groups (eight mice/group): Sham surgery with standard environment (SE) or enriched environment (EE) or spare nerve injury (SNI) with SE or EE. Mice in EE groups had access to an exercise wheel. Mothers were evaluated for mechanical sensitivity with Von Frey filaments and for exercise performance with computerized running wheels. Mice were impregnated 2 weeks after the initiation of EE. Pups were weighed and measured for length at birth and evaluated for negative geotaxis, righting, forelimb grasping, rooting, and crawling at 3 days postpartum and for crawling at 6 days postpartum. Following euthanasia, mothers' frontal cortexes were analyzed for selected neuropeptides. After exercise exposure, only SNI-SE females remained neuropathic. Exercise levels were similar between EE groups. Some brain neuropeptides (endorphins, enkephalins, and oxytocin) from SNI females showed significant differences with exercise. Number of pups was significantly smaller in the SNI-SE group. Significantly more pups died at birth in the SNI-SE group, but pup behavior tests (except righting) were similar across groups. Exercise can reduce neuropathic pain in pregnant mice. Neuropathic pain does not impact motor neurodevelopment of mice pups but does appear to affect litter size and neonatal mortality.


Plasmodium falciparum malaria in children aged 0-2 years: the role of foetal haemoglobin and maternal antibodies to two asexual malaria vaccine candidates (MSP3 and GLURP).
Children below six months are reported to be less susceptible to clinical malaria. Maternally derived antibodies and foetal haemoglobin are important putative protective factors. We examined antibodies to Plasmodium falciparum merozoite surface protein 3 (MSP3) and glutamate-rich protein (GLURP), in children in their first two years of life in Burkina Faso and their risk of malaria. A cohort of 140 infants aged between four and six weeks was recruited in a stable transmission area of south-western Burkina Faso and monitored for 24 months by active and passive surveillance. Malaria infections were detected by examining blood smears using light microscopy. Enzyme-linked immunosorbent assay was used to quantify total Immunoglobulin G to Plasmodium falciparum antigens MSP3 and two regions of GLURP (R0 and R2) on blood samples collected at baseline, three, six, nine, 12, 18 and 24 months. Foetal haemoglobin and variant haemoglobin fractions were measured at the baseline visit using high pressure liquid chromatography. A total of 79.6% of children experienced one or more episodes of febrile malaria during monitoring. Antibody titres to MSP3 were prospectively associated with an increased risk of malaria while antibody responses to GLURP (R0 and R2) did not alter the risk. Antibody titres to MSP3 were higher among children in areas of high malaria risk. Foetal haemoglobin was associated with delayed first episode of febrile malaria and haemoglobin CC type was associated with reduced incidence of febrile malaria. We did not find any evidence of association between titres of antibodies to MSP3, GLURP-R0 or GLURP-R2 as measured by enzyme-linked immunosorbent assay and early protection against malaria, although anti-MSP3 antibody titres may reflect increased exposure to malaria and therefore greater risk. Foetal haemoglobin was associated with protection against febrile malaria despite the study limitations and its role is therefore worthy further investigation.


Health savings accounts: early estimates of national take-up.
The 2003 Medicare Prescription Drug, Improvement, and Modernization Act (MMA) approved tax-advantaged health savings accounts (HSAs) for certain high-deductible health insurance plans. We predict that MMA could lead to approximately 3.2 million HSA contracts among Americans ages 19-64 who are not students, not enrolled in public health insurance plans, and not eligible for group coverage as a dependent. We simulate the effect of several additional tax subsidies for HSAs. We predict that the Bush administration's refundable tax-credit proposal would double HSA take-up and reduce the number of uninsured people by 2.9 million, at an annual cost of $8.1 billion.


The initial assessment of expert panel performance in core hospitals for cancer genomic medicine in Japan.
Since June 2019, cancer genomic profiling (CGP) tests have been reimbursed by the National Health Insurance system in Japan, with restrictions for government-designated hospitals with a molecular tumor board composed of multidisciplinary specialists, known as an expert panel (EP). The standardization of EPs is a critical challenge for implementing precision oncology in the clinical setting. Data on consecutive cases who underwent the CGP tests at 11 core hospitals between June 2019 and January 2020 were collected. We evaluated the proportions of cases that received genomically matched treatments, including investigational new drugs (INDs) based on CGP results, and/or for which genetic counseling was recommended. Two simulated cases were annotated by each EP. The annotated reports were then centrally assessed. Each EP mainly discussed the applicability to genomically matched treatments and the necessity of performing genetic counseling. A pre-review of the report by key members in each EP reportedly made the EP conference more interactive and efficient, and thereby saved time. A total of 747 cases underwent CGP tests, 28 cases (3.7%) received genomically matched treatment, and 17 cases (2.3%) were referred for genetic counseling. Annotated reports for the simulated cases varied across the EPs, particularly the number of recommended IND trials, which seemed to be associated with the actual number of participants in IND trials. This investigation provides reference data for the application of precision oncology in a clinical setting. Further investigations on the standardization of clinical annotations are warranted.


Lymph nodes as barriers to T-cell rejuvenation in aging mice and nonhuman primates.
In youth, thymic involution curtails production of new naïve T cells, placing the onus of T-cell maintenance upon secondary lymphoid organs (SLO). This peripheral maintenance preserves the size of the T-cell pool for much of the lifespan, but wanes in the last third of life, leading to a dearth of naïve T cells in blood and SLO, and contributing to suboptimal immune defense. Both keratinocyte growth factor (KGF) and sex steroid ablation (SSA) have been shown to transiently increase the size and cellularity of the old thymus. It is less clear whether this increase can improve protection of old animals from infectious challenge. Here, we directly measured the extent to which thymic rejuvenation benefits the peripheral T-cell compartment of old mice and nonhuman primates. Following treatment of old animals with either KGF or SSA, we observed robust rejuvenation of thymic size and cellularity, and, in a reporter mouse model, an increase in recent thymic emigrants (RTE) in the blood. However, few RTE were found in the spleen and even fewer in the lymph nodes, and SSA-treated mice showed no improvement in immune defense against West Nile virus. In parallel, we found increased disorganization and fibrosis in old LN of both mice and nonhuman primates. These results suggest that SLO defects with aging can negate the effects of successful thymic rejuvenation in immune defense.


Dok-1 and Dok-2 Are Required To Maintain Herpes Simplex Virus 1-Specific CD8+ T Cells in a Murine Model of Ocular Infection.
Dok-1 and Dok-2 negatively regulate responses downstream of several immune receptors in lymphoid and myeloid cells. Recent evidence showed that Dok proteins are essential in the formation of memory CD8+ T cells to an exogenous epitope expressed by vaccinia virus; however, the importance of Dok-1 and Dok-2 in the control of viral infection is unknown. Here, we investigated the role of Dok proteins in modulating the immune response against herpes simplex virus 1 (HSV-1) in a mouse model of ocular infection. During acute infection, viral titers in the eye were similar in wild-type (WT) and Dok-1 and Dok-2 double-knockout (DKO) mice, and the percentages of infiltrating leukocytes were similar in DKO and WT corneas and trigeminal ganglia (TG). DKO mice exhibited a diminished CD8+ T cell response to the immunodominant HSV-1 glycoprotein B (gB) epitope in the spleen and draining lymph nodes compared to WT mice during acute infection. Remarkably, gB-specific CD8+ T cells almost completely disappeared in the spleens of DKO mice during latency, and the reduction of CD8+ effector memory T (Tem) cells was more severe than that of CD8+ central memory T (Tcm) cells. The percentage of gB-specific CD8+ T cells in TG during latency was also dramatically reduced in DKO mice; however, they were phenotypically similar to those from WT mice. In ex vivo assays, reactivation was detected earlier in TG cultures from infected DKO versus WT mice. Thus, Dok-1 and Dok-2 promote survival of gB-specific CD8+ T cells in TG latently infected with HSV-1.IMPORTANCE HSV-1 establishes lifelong latency in sensory neurons of trigeminal ganglia (TG). In humans, HSV-1 is able to sporadically reactivate from latently infected neurons and establish a lytic infection at a site to which the neurons project. Most herpetic disease in humans is due to reactivation of HSV-1 from latency rather than to primary acute infection. CD8+ T cells are thought to play an important role in controlling recurrent infections. In this study, we examined the involvement of Dok-1 and Dok-2 signaling proteins in the control of HSV-1 infection. We provide evidence that Dok proteins are required to maintain a CD8+ T cell response against HSV-1 during latency-especially CD8+ Tem cells-and that they negatively affect HSV-1 reactivation from latency. Elucidating Dok-mediated mechanisms involved in the control of HSV-1 reactivation from latency might contribute to the development of therapeutic strategies to prevent recurrent HSV-1-induced pathology.


High-performance guided-wave asynchronous heralded single-photon source.
We report on a guided-wave asynchronous heralded single-photon source based on the creation of nondegenerate photon pairs by spontaneous parametric downconversion in a periodically poled lithium niobate wave-guide. We show that, by use of the signal photon at 1310 nm as a trigger, a gated detection process permits announcement of the arrival of single photons at 1550 nm at the output of a single-mode optical fiber with a high probability of 0.37. At the same time the multiphoton emission probability is reduced by a factor of 10 compared with Poissonian light sources. Furthermore, the model we have developed to calculate those figures of merit is shown to be accurate. This study can therefore serve as a paradigm for the conception of new quantum communication and computation networks.


Mapping the binding sites of human erythrocyte ankyrin for the anion exchanger and spectrin.
This report describes initial characterization of the binding sites of ankyrin for spectrin and the anion exchanger using defined subfragments isolated from purified ankyrin domains. The spectrin-binding domain of ankyrin is comprised of two subdomains: an acidic, proline-rich region (pI = 4) involving the amino-terminal 80 residues from 828 to 908 and a basic region (pI = 8.8) that extends from 898 to 1386. The amino-terminal 70 amino acids of the spectrin-binding domain are critical for association with spectrin, since a subfragment missing this region is only 5% as active as the intact domain in displacing binding of spectrin to inside-out membrane vesicles, while deletion of the first 38 residues of the acidic domain results in a 10-fold reduction in activity. The anion exchanger-binding site is confined to an 89-kDa domain that was isolated and characterized as a globular molecule with approximately 30% alpha-helical configuration. A subfragment of the 89-kDa domain extending from residues 403 to 779 (or possibly 740) retains ability to associate with the anion exchanger. The 89-kDa domain is comprised of a series of tandem repeats of 33 amino acids that extend from residues 35 to 778 (Lux, S., John, K., and Bennett, V. (1990) Nature 344, 36-42). The activity of residues 403-779 demonstrates that the 33-amino acid repeats of the 89-kDa domain are responsible for association between ankyrin and the anion exchanger. The 33-amino acid repeating sequence of ankyrin represents an ancient motif also found in proteins of Drosophila, yeast, and Caenor habditis elegans. The finding that the 33-amino acid repeating sequence is involved in interaction with the anion exchanger implies that this motif may perform a role in molecular recognition in diverse proteins.


A Randomized Trial of Caspofungin vs Triazoles Prophylaxis for Invasive Fungal Disease in Pediatric Allogeneic Hematopoietic Cell Transplant.
Children and adolescents undergoing allogeneic hematopoietic cell transplantation (HCT) are at high risk for invasive fungal disease (IFD). This multicenter, randomized, open-label trial planned to enroll 560 children and adolescents (3 months to <21 years) undergoing allogeneic HCT between April 2013 and September 2016. Eligible patients were randomly assigned to antifungal prophylaxis with caspofungin or a center-specific comparator triazole (fluconazole or voriconazole). Prophylaxis was administered from day 0 of HCT to day 42 or discharge. The primary outcome was proven or probable IFD at day 42 as adjudicated by blinded central review. Exploratory analysis stratified this evaluation by comparator triazole. A planned futility analysis demonstrated a low rate of IFD in the comparator triazole arm, so the trial was closed early. A total of 290 eligible patients, with a median age of 9.5 years (range 0.3-20.7), were randomized to caspofungin (n = 144) or a triazole (n = 146; fluconazole, n = 100; voriconazole, n = 46). The day 42 cumulative incidence of proven or probable IFD was 1.4% (95% confidence interval [CI], 0.3%-5.4%) in the caspofungin group vs 1.4% (95% CI, 0.4%-5.5%) in the triazole group (P = .99, log-rank test). When stratified by specific triazole, there was no significant difference in proven or probable IFD at day 42 between caspofungin vs fluconazole (1.0%, 95% CI, 0.1%-6.9%, P = .78) or caspofungin vs voriconazole (2.3%, 95% CI, 0.3%-15.1%, P = .69). In pediatric HCT patients, prophylaxis with caspofungin did not significantly reduce the cumulative incidence of early proven or probable IFD compared with triazoles. Future efforts to decrease IFD-related morbidity and mortality should focus on later periods of risk. NCT01503515.


Transmission wavefront shearing interferometry for photoelastic materials.
A general analysis and experimental validation of transmission wavefront shearing interferometry for photoelastic materials are presented. These interferometers applied to optically isotropic materials produce a single interference pattern related to one phase term, but when applied to photoelastic materials, they produce the sum of two different interference patterns with phase terms that are the sum and difference, respectively, of two stress-related phase terms. The two stress-related phase terms may be separated using phase shifting and polarization optics. These concepts are experimentally demonstrated using coherent gradient sensing in full field for a compressed polycarbonate plate with a V-shaped notch with good agreement with theoretical data. The analysis may be applied to any wavefront shearing interferometer by modifying parameters describing the wavefront shearing distance.


The reproducibility of limb blood flow measurements in human volunteers at rest and after exercise by using mercury-in-Silastic strain gauge plethysmography under standardized conditions.
A study to assess the reproducibility of limb blood flow measurements by venous occlusion plethysmography using mercury-in-Silastic strain gauges was performed in six normal volunteers under standardized conditions. Using this technique forearm and calf blood flow were measured at rest and between 2 and 3 min after submaximal exercise on six separate occasions over a 1 month period. The mean coefficient of variation for resting forearm blood flow was 10.5% (range 7.8-15.6%). The mean coefficient of variation for resting calf blood flow was 11.5% (range 7.4-14.2%). The mean coefficient of variation for post-exercise calf blood flow was 13% (range 11.5-16.4%). The results suggest that limb blood flow measurement by this technique may be useful in studies where serial measurements are required.