Flavobacterium chilense sp. nov. and Flavobacterium araucananum sp. nov., isolated from farmed salmonid fish.
Three Gram-staining-negative non-endospore-forming strains were isolated from farmed fish in Chile: one (LM-09-Fp(T)) from a rainbow trout (Oncorhynchus mykiss) and the others (LM-19-Fp(T) and LM-20-Fp) from two Atlantic salmon (Salmo salar). Phylogenetic analyses based on 16S rRNA gene sequences indicated that all three isolates belonged to the genus Flavobacterium. In these analyses, strain LM-09-Fp(T) appeared most closely related to the type strains of Flavobacterium chungangense (98.5 % sequence similarity), Flavobacterium glaciei (98.2 %), Flavobacterium aquidurense (97.6 %), Flavobacterium saccharophilum (97.6 %) and Flavobacterium hercynium (97.6 %). The 16S rRNA gene sequences of strains LM-19-Fp(T) and LM-20-Fp were found to be identical and most similar to the corresponding sequences of the type strains of Flavobacterium aquidurense (98.6 %), Flavobacterium frigidimaris (98.5 %), Flavobacterium hercynium (97.9 %), Flavobacterium saccharophilum (97.7 %) and Flavobacterium pectinovorum (97.7 %). For each of the three novel strains, menaquinone (MK-6) was the predominant respiratory quinone and the major compounds in the polar lipid profile were phosphatidylethanolamine, an unidentified aminolipid, phosphatidylserine and two or three unknown lipids. The fatty acid profile of each strain, which comprised major amounts of iso-C(15:0), C(15:0) and summed feature 3 (C(16:1)ω7c and/or iso-C(15:0) 2-OH) as well as smaller amounts of various hydroxylated fatty acids (e.g. iso-C(16:0) 3-OH, iso-C(17:0) 3-OH, C(16:0) 3-OH and C(15:0) 3-OH), indicated that each belonged to the genus Flavobacterium. Based on their physiological and biochemical characteristics and the results of DNA-DNA hybridizations, which showed relatively low levels of relatedness between the novel strains and the most closely related Flavobacterium species, strain LM-09-Fp(T) ( = LMG 26360(T) = CCM 7940(T)) represents a novel species within the genus Flavobacterium, for which the name Flavobacterium chilense sp. nov. is proposed, and strains LM-19-Fp(T) ( = LMG 26359(T) = CCM 7939(T)) and LM-20-Fp ( = LMG 26331) represent a second novel species within the same genus, for which the name Flavobacterium araucananum sp. nov. is proposed.


Neural and behavioral changes driven by observationally-induced hypoalgesia.
Observing successful pain treatment in others can induce anticipatory neural processes that, in turn, relieve pain. Previous studies have suggested that social learning and observation influence placebo hypoalgesia. Here, we used electroencephalography (EEG) to determine the neurophysiological changes associated with pain relief acquired through the observation. Thirty-one participants observed a demonstrator undergo painful heat stimulations paired with a "control" cream and non-painful ones paired with a "treatment" cream, which actually were both Vanicreams. After their observation, the participants then received the same creams and stimulations. We found that the treatment cream led to lower self-reported pain intensity ratings than the control cream. Anticipatory treatment cues elicited smaller P2 in electrodes F1, Fz, FC1, and FCz than the control condition. The P2 component localization indicated a higher current density in the right middle frontal gyrus, a region associated with attentional engagement. In placebo responders, the sensorimotor cortex activity captured in electrodes C3, Cz, and C4 indicated that hypoalgesia was positively correlated with resting state peak alpha frequency (PAF). These results suggest that observationally-induced placebo hypoalgesia may be driven by anticipatory mechanisms that modulate frontal attentional processes. Furthermore, resting state PAF could serve as a predictor of observationally-induced hypoalgesia.


Three-dimensional coupled-wave theory for the guided mode resonance in photonic crystal slabs: TM-like polarization.
A general coupled-wave theory is presented for the guided resonance in photonic crystal (PhC) slabs with TM-like polarization. Numerical results based on our model are presented with finite-difference time-domain validations. The proposed analysis facilitates comprehensive understanding of the physics of guided resonance in PhC slabs and provides guidance for its applications.


Pathology turnaround time in pediatric oncology: a tool to prepare patients and families for the diagnostic waiting period.
There is little data on the amount of time patients and families typically wait for pathology results when pediatric malignancy is suspected. The purpose of this study was to determine the average waiting period after diagnostic intervention for pediatric cancer. Pathology reports were reviewed for pediatric patients who had their initial diagnosis and were followed in the Division of Hematology/Oncology from 2007 through 2010. The average turnaround time (TAT) for all pathology (n=266) was 6.9 days. The TAT for pathology results according to diagnosis was 10.1 days for CNS tumors (n=59), 9.7 days for sarcomas (n=40), 5.4 days for lymphomas (n=31), 5.4 days for neuroblastoma (n=13), 7.3 days for kidney tumors (n=11), 7.2 days for thyroid tumors (n=7), 9.4 days for ovarian tumors (n=7), 7.0 days for schwannomas/neurofibromas (n=5), 5.7 days for testicular tumors (n=3), 5.0 days for hepatoblastoma (n=3), and 7.0 days for nasopharyngeal carcinomas (n=2). Overall the TAT for leukemia was 3.1 days (n=76), with diagnosis by flow cytometry taking 1.2 days and results by bone marrow biopsy taking 4.0 days. The TAT for pediatric oncology pathology after diagnostic intervention varies according to diagnosis. The hope is that this information will better prepare patients and families for the agonizing waiting period associated with diagnosis.


The pleiotropy of telomerase against cell death.
The end of eukaryotic genomic DNA is capped by a specialized structure called as "telomere", which consists of the repetitive array of nucleotide sequence, TTAGGG, in humans and mice, and a variety of binding proteins. Telomerase is a ribonucleoprotein (RNP) complex responsible for the elongation of telomeres to maintain the genomic integrity, and is composed of telomerase reverse transcriptase (TERT), telomerase RNA component (TERC), and their associated factors regulating the catalytic activity of telomerase. Although it is now apparent that telomerase protects cells from apoptosis via the maintenance of genomic integrity by stabilizing telomeres, our understanding for the physiological role of telomerase is yet far from completion, and emerging evidence suggests that telomerase has additional extratelomeric roles in mediating cell survival and anti-apoptotic functions against various cytotoxic stresses. Here we summarize and discuss how telomerase and telomeres are involved in mediating cellular protection against apoptosis.


Regulation of skeletal muscle regeneration by CCR2-activating chemokines is directly related to macrophage recruitment.
Muscle regeneration requires CC chemokine receptor 2 (CCR2) expression on bone marrow-derived cells; macrophages are a prominent CCR2-expressing cell in this process. CCR2-/- mice have severe impairments in angiogenesis, macrophage recruitment, and skeletal muscle regeneration following cardiotoxin (CTX)-induced injury. However, multiple chemokines activate CCR2, including monocyte chemotactic proteins (MCP)-1, -3, and -5. We hypothesized that MCP-1 is the chemokine ligand that mediates the impairments present in CCR2-/- mice. We examined muscle regeneration, capillary density, and cellular recruitment in MCP-1-/- and CCR2-/- mice following injury. Muscle regeneration and adipocyte accumulation, but not capillary density, were significantly impaired in MCP-1-/- compared with wild-type (WT) mice; however, muscle regeneration and adipocyte accumulation impairments were not as severe as observed in CCR2-/- mice. Although tissue levels of MCP-5 were elevated in MCP-1-/- mice compared with WT, the administration of MCP-5 neutralizing antibody did not alter muscle regeneration in MCP-1-/- mice. While neutrophil accumulation after injury was similar in all three mouse strains, macrophage recruitment was highest in WT mice, intermediate in MCP-1-/- mice, and severely impaired in CCR2-/- mice. In conclusion, while the absence of MCP-1 resulted in impaired macrophage recruitment and muscle regeneration, MCP-1-/- mice exhibit an intermediate phenotype compared with CCR2-/- mice. Intermediate macrophage recruitment in MCP-1-/- mice was associated with similar capillary density to WT, suggesting that fewer macrophages may be needed to restore angiogenesis vs. muscle regeneration. Finally, other chemokines, in addition to MCP-1 and MCP-5, may activate CCR2-dependent regenerative processes resulting in an intermediate phenotype in MCP-1-/- mice.


The independent prospective associations of activity intensity and dietary energy density with adiposity in young adolescents.
There is limited evidence on the prospective association of time spent in activity intensity (sedentary (SED), moderate (MPA) or vigorous (VPA) physical activity) and dietary intake with adiposity indicators in young people. This study aimed to assess associations between (1) baseline objectively measured activity intensity, dietary energy density (DED) and 4-year change in adiposity and (2) 4-year change in activity intensity/DED and adiposity at follow-up. We conducted cohort analyses including 367 participants (10 years at baseline, 14 years at follow-up) with valid data for objectively measured activity (Actigraph), DED (4-d food diary), anthropometry (waist circumference (WC), %body fat (%BF), fat mass index (FMI), weight status) and covariates. Linear and logistic regression models were fit, including adjustment for DED and moderate-to-vigorous physical activity. Results showed that baseline DED was associated with change in WC (β for 1kJ/g difference: 0·71; 95% CI 0·26, 1·17), particularly in boys (1·26; 95% CI 0·41, 2·16 v. girls: 0·26; 95% CI -0·34, 0·87), but not with %BF, FMI or weight status. In contrast, baseline SED, MPA or VPA were not associated with any of the outcomes. Change in DED was negatively associated with FMI (β for 1kJ/g increase: -0·86; 95% CI -1·59, -0·12) and %BF (-0·86; 95% CI -1·25, -0·11) but not WC (-0·27; 95% CI -1·02, 0·48). Change in SED, MPA and VPA did not predict adiposity at follow-up. In conclusion, activity intensity was not prospectively associated with adiposity, whereas the directions of associations with DED were inconsistent. To inform public health efforts, future studies should continue to analyse longitudinal data to further understand the independent role of different energy-balance behaviours in changes in adiposity in early adolescence.


Folate transport in intestinal brush border membrane: involvement of essential histidine residue(s).
We examined the possible existence of histidine residue(s) in the folate transporter of rabbit intestine. This was done with use of the histidine-specific reagent diethyl pyrocarbonate (DEPC) and purified intestinal brush-border-membrane vesicles. DEPC caused significant concentration- and time-dependent inhibition of folic acid transport. The inhibition was only seen when transport was examined in vesicles incubated in buffer at pH 5.2 and not in those incubated in buffer at pH 7.4. The addition of unlabelled folic acid to vesicle suspension before treatment with DEPC (2.5 mM) led to a significant (P < 0.01) protection (84%) against the inhibition of folic acid transport. Treating vesicles pretreated with DEPC (2.5 mM) with reducing reagents (dithiothreitol, 2-mercaptoethanol and 2,3-dimercaptopropanol, all at a final concentration of 10 mM) did not reverse the inhibitory effect of DEPC on folic acid transport. On the other hand, treating the DEPC-pretreated vesicles with hydroxylamine (140 mM) led to a significant reversal (P < 0.01) (54%) of the inhibition of folic acid transport. The inhibitory effect of DEPC on carrier-mediated folic acid transport was found to be mediated through a decrease in the Vmax. (i.e. a decrease in the number and/or activity) of the carriers and an increase in the apparent Km (i.e. a decrease in their affinity), classifying the effect as a mixed-type inhibition. These results demonstrate the existence of critical histidine residue(s) in the intestinal brush-border-membrane folate transporter which is essential for its interaction with, and transport of, the vitamin. These findings also suggest that the histidine residue(s) is located at (or near) the substrate-binding site.


Association of psychological flexibility with engagement in pulmonary rehabilitation following an acute exacerbation of chronic obstructive pulmonary disease.
This study aimed to investigate (a) the association between psychological flexibility and engagement in pulmonary rehabilitation within 8 weeks following hospitalisation for an acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and (b) how psychological (in)flexibility presents in this context. A mixed-methods study was conducted. Psychological flexibility during an AECOPD was assessed using The Acceptance and Action Questionnaire-II (AAQ-II) (n = 41) and the Engaged Living Scale (ELS) (n = 40). Engagement in post-AECOPD pulmonary rehabilitation was then recorded. Twenty-three patients also participated in cognitive interviews. Psychological flexibility was associated with a greater chance of accepting a pulmonary rehabilitation referral following an AECOPD. Small numbers prohibited analysis on attendance or completion. An AAQ-II score of 11 translated to a 60 (37-82)% probability of accepting a referral to pulmonary rehabilitation and an ELS score of 73 was associated with a 68 (46-91)% probability of accepting. Four themes were extracted from interviews: (1) family values, (2) self as abnormal, (3) 'can't do anything' versus 'I do what I can' and (4) disability, and related emotions, as barriers to action. Randomised clinical trials are needed to evaluate interventions designed to increase psychological flexibility (i.e. acceptance and commitment therapy) to support acceptance of pulmonary rehabilitation post-AECOPD.


Sensitive method for the kinetic measurement of trace species in liquids using cavity enhanced absorption spectroscopy with broad bandwidth supercontinuum radiation.
A novel spectrometer for the rapid and sensitive detection of liquid phase analytes at trace concentrations is presented. Broad bandwidth supercontinuum radiation was coupled into a linear optical cavity incorporating an intracavity liquid-sample cuvette. Cavity enhanced absorption spectra of trace species covering more than 300 nm were acquired on time scales of milliseconds. Single shot acquisition times of 10-50 ms are demonstrated here. The effective absorption path length exceeds 2 m in sample volumes measuring 2.7 mL. A key feature of the instrument is that it can be calibrated using cavity ring-down spectroscopy without the requirement of changing the optical alignment. The sensitivity of the instrument is exemplified by measurements of trace concentrations of dye molecules and nickel sulfate dissolved in water. A minimum detectable absorption coefficient of 9.1 x 10(-7) cm(-1) Hz(-1/2) at 550 nm was obtained. The capability to capture broad bandwidth absorption spectra on short time scales permits kinetic studies of liquid phase reactions. We demonstrate this by recording the oscillatory behavior of a Belousov-Zhabotinsky reaction.


The eyes and ears are visual indicators of attention in domestic horses.
Sensitivity to the attentional states of others has adaptive advantages, and in social animals, attending to others is important for predator detection, as well as a pre-requisite for normal social functioning and more complex socio-cognitive abilities. Despite widespread interest in how social species perceive attention in others, studies of non-human animals have been inconclusive about the detailed cues involved. Previous work has focused on head and eye direction, overlooking the fact that many mammals have obvious and mobile ears that could act as a visual cue to attention. Here we report that horses use the head orientation of a conspecific to locate food, but that this ability is disrupted when parts of the face (the eyes and ears) are covered up with naturalistic masks. The ability to correctly judge attention also interacted with the identity of the model horse, suggesting that individual differences in facial features may influence the salience of cues. Our results indicate that a combination of head orientation with facial expression, specifically involving both the eyes and ears, is necessary for communicating social attention. These findings emphasise that in order to understand how attention is communicated in non-human animals, it is essential to consider a broad range of cues.


Leucodepletion for transmissible spongiform encephalopathies.
Transmissible spongiform encephalopathies (TSEs) have been recognised around the world for many years. Creutzfeldt-Jakob disease (CJD), one of the human forms of TSE, has been studied widely and thus far has not proved a great threat to human health. The emergence of two new TSEs--bovine spongiform encephalopathy (BSE) in cattle and variant Creutzfeldt-Jakob disease (vCJD) in humans in the UK--has caused great concern. BSE has had an economic impact and vCJD is a threat to human health. It has been shown that these two diseases are caused by the same prion agent and are linked. Research indicates that vCJD behaves differently to CJD and there is strong evidence to suggest that vCJD is present in lymphoid tissues and B lymphocytes, which presents a theoretical risk that it may be transmitted by transfusion of blood and blood products. To minimise/prevent this risk, the UK government has decided that plasma should be sourced from abroad and has instructed the National Blood Service to leucodeplete all blood and blood products, at a cost of 70 million pounds per annum, although it is not known if this will remove this risk.


The efficacy of Artecoll injections for the augmentation of nipple projection in breast reconstruction.
Various techniques have been used in an attempt to achieve long-term nipple projection following nipple-areolar reconstruction. A common setback, however, is the diminution of projection overtime; this phenomenon is particularly evident following implant-based breast reconstruction. Artecoll may be suitable for injection into the nipple complex to maintain permanent, 3-dimensional projection. Artecoll is an injectable substance that is biocompatible and immunologically inert and resists degradation in vivo. The purpose of this study was thus to prospectively evaluate the efficacy of Artecoll (polymethylmethacrylate microspheres suspended in 3.5% denatured bovine collagen with 0.3% lidocaine) in obtaining and maintaining nipple projection following postmastectomy, nipple-areolar reconstruction. A prospective, clinical trial was performed. Consecutive patients deemed to have inadequate nipple projection at least 6 months following "C-V flap" or "modified-skate flap" reconstruction were identified. Only women who had postmastectomy reconstruction with tissue expanders and implants were considered eligible for participation. Artecoll was injected under the nipple at 2 time points: baseline and 3 months. Calipers were used to measure nipple projection preinjection and postinjection at baseline, 3, 6, and 9 months. Thirty-three nipples were injected in 23 patients. There were no adverse events. Prior to injection, mean nipple projection was 1.33 +/- 1.0 mm. The mean increase in projection over the 9-month study period was both clinically and statistically significant (1.60 +/- 1.24 mm; P <.001). A history of prior irradiation was a significant negative predictor of final nipple projection (P = .012). Artecoll injection is both feasible and effective in increasing and maintaining nipple projection in the setting of implant-based breast reconstruction.


Aligned carbon nanotube array stiffness from stochastic three-dimensional morphology.
The landmark theoretical properties of low dimensional materials have driven more than a decade of research on carbon nanotubes (CNTs) and related nanostructures. While studies on isolated CNTs report behavior that aligns closely with theoretical predictions, studies on cm-scale aligned CNT arrays (>10(10) CNTs) oftentimes report properties that are orders of magnitude below those predicted by theory. Using simulated arrays comprised of up to 10(5) CNTs with realistic stochastic morphologies, we show that the CNT waviness, quantified via the waviness ratio (w), is responsible for more than three orders of magnitude reduction in the effective CNT stiffness. Also, by including information on the volume fraction scaling of the CNT waviness, the simulation shows that the observed non-linear enhancement of the array stiffness as a function of the CNT close packing originates from the shear and torsion deformation mechanisms that are governed by the low shear modulus (∼1 GPa) of the CNTs.


Management, prognosis and predictors of unfavourable outcomes in patients newly hospitalized for transient ischemic attack: a real-world investigation from Italy.
Understanding the gap between evidence-based recommendations and real-world management is important to inform priority setting and health service planning. The 7,776 residents in the Italian Lombardy Region who were newly hospitalized for transient ischemic attack (TIA) during 2008-2009 entered into the cohort and were followed until 2012. Exposure to medical care including selected drugs, diagnostic procedures and laboratory tests was recorded. A composite outcome was employed taking into account all-cause death and hospitalization for stroke and acute myocardial infarction. A multivariable proportional hazards model was fitted to estimate hazard ratio, and 95% confidence intervals (CI), for the exposure-outcome association. During the first year after discharge, 8.6, 49.7 and 48.5% of patients did not use any drugs, diagnostic procedures and laboratory tests respectively. Patients exposed to medical care had 59% reduced risk (95% CI, 50 to 66%) with respect to those who did not use any of these services. Although the Italian National Health System supplies universal coverage for healthcare, several TIA patients receive suboptimal care. Systematic improvements are necessary in order to improve patient outcomes.


Source apportionment of circum-Arctic atmospheric black carbon from isotopes and modeling.
Black carbon (BC) contributes to Arctic climate warming, yet source attributions are inaccurate due to lacking observational constraints and uncertainties in emission inventories. Year-round, isotope-constrained observations reveal strong seasonal variations in BC sources with a consistent and synchronous pattern at all Arctic sites. These sources were dominated by emissions from fossil fuel combustion in the winter and by biomass burning in the summer. The annual mean source of BC to the circum-Arctic was 39 ± 10% from biomass burning. Comparison of transport-model predictions with the observations showed good agreement for BC concentrations, with larger discrepancies for (fossil/biomass burning) sources. The accuracy of simulated BC concentration, but not of origin, points to misallocations of emissions in the emission inventories. The consistency in seasonal source contributions of BC throughout the Arctic provides strong justification for targeted emission reductions to limit the impact of BC on climate warming in the Arctic and beyond.


Integrative Computational Network Analysis Reveals Site-Specific Mediators of Inflammation in Alzheimer's Disease.
Alzheimer's disease (AD) is a major neurodegenerative disease and is one of the most common cause of dementia in older adults. Among several factors, neuroinflammation is known to play a critical role in the pathogenesis of chronic neurodegenerative diseases. In particular, studies of brains affected by AD show a clear involvement of several inflammatory pathways. Furthermore, depending on the brain regions affected by the disease, the nature and the effect of inflammation can vary. Here, in order to shed more light on distinct and common features of inflammation in different brain regions affected by AD, we employed a computational approach to analyze gene expression data of six site-specific neuronal populations from AD patients. Our network based computational approach is driven by the concept that a sustained inflammatory environment could result in neurotoxicity leading to the disease. Thus, our method aims to infer intracellular signaling pathways/networks that are likely to be constantly activated or inhibited due to persistent inflammatory conditions. The computational analysis identified several inflammatory mediators, such as tumor necrosis factor alpha (TNF-a)-associated pathway, as key upstream receptors/ligands that are likely to transmit sustained inflammatory signals. Further, the analysis revealed that several inflammatory mediators were mainly region specific with few commonalities across different brain regions. Taken together, our results show that our integrative approach aids identification of inflammation-related signaling pathways that could be responsible for the onset or the progression of AD and can be applied to study other neurodegenerative diseases. Furthermore, such computational approaches can enable the translation of clinical omics data toward the development of novel therapeutic strategies for neurodegenerative diseases.


First report of Mycobacterium bovis DNA in human remains from the Iron Age.
Tuberculosis has plagued humankind since prehistoric times, as is evident from characteristic lesions on human skeletons dating back to the Neolithic period. The disease in man is due predominantly to infection with either Mycobacterium tuberculosis or Mycobacterium bovis, both members of the M. tuberculosis (MTB) complex. A number of studies have shown that when conditions permit, surviving mycobacterial DNA may be amplified from bone by PCR. Such ancient DNA (aDNA) analyses are subject to stringent tests of authenticity and, when feasible, are invariably limited by DNA fragmentation. Using PCRs based on single-nucleotide polymorphic loci and regions of difference (RDs) in the MTB complex, a study was made of five Iron Age individuals with spinal lesions recovered from the cemetery of Aymyrlyg, South Siberia. A sensitive screening PCR for MTB complex mycobacteria was positive in four out of the five cases. Genotyping evidence indicated that all four cases were due to infection with M. bovis rather than M. tuberculosis and the data were consistent with the proposed phylogenetic model of the MTB complex. This is believed to be the first report of M. bovis causing Pott's disease in archaeological human remains. The study shows that genotyping of ancestral strains of MTB complex mycobacteria from contexts of known date provides information which allows the phylogeny of the model to be tested. Moreover, it shows that loss of DNA from RD4, which defines classic M. bovis, had already occurred from the genome over 2000 years before the present.


How countries cope with competing demands and expectations: perspectives of different stakeholders on priority setting and resource allocation for health in the era of HIV and AIDS.
Health systems have experienced unprecedented stress in recent years, and as yet no consensus has emerged as to how to deal with the multiple burden of disease in the context of HIV and AIDS and other competing health priorities. Priority setting is essential, yet this is a complex, multifaceted process. Drawing on a study conducted in five African countries, this paper explores different stakeholders' perceptions of health priorities, how priorities are defined in practice, the process of resource allocation for HIV and Health and how different stakeholders perceive this. A sub-analysis was conducted of selected data from a wider qualitative study that explored the interactions between health systems and HIV and AIDS responses in five sub-Saharan countries (Burkina Faso, the Democratic Republic of Congo, Ghana, Madagascar and Malawi). Key background documents were analysed and semi-structured interviews (n = 258) and focus group discussions (n = 45) were held with representatives of communities, health personnel, decision makers, civil society representatives and development partners at both national and district level. Health priorities were expressed either in terms of specific health problems and diseases or gaps in service delivery requiring a strengthening of the overall health system. In all five countries study respondents (with the exception of community members in Ghana) identified malaria and HIV as the two top health priorities. Community representatives were more likely to report concerns about accessibility of services and quality of care. National level respondents often referred to wider systemic challenges in relation to achieving the Millennium Development Goals (MDGs). Indeed, actual priority setting was heavily influenced by international agendas (e.g. MDGs) and by the ways in which development partners were supporting national strategic planning processes. At the same time, multi-stakeholder processes were increasingly used to identify priorities and inform sector-wide planning, whereby health service statistics were used to rank the burden of disease. However, many respondents remarked that health system challenges are not captured by such statistics.In all countries funding for health was reported to fall short of requirements and a need for further priority setting to match actual resource availability was identified. Pooled health sector funds have been established to some extent, but development partners' lack of flexibility in the allocation of funds according to country-generated priorities was identified as a major constraint. Although we found consensus on health priorities across all levels in the study countries, current funding falls short of addressing these identified areas. The nature of external funding, as well as programme-specific investment, was found to distort priority setting. There are signs that existing interventions have had limited effects beyond meeting the needs of disease-specific programmes. A need for more comprehensive health system strengthening (HSS) was identified, which requires a strong vision as to what the term means, coupled with a clear strategy and commitment from national and international decision makers in order to achieve stated goals. Prospective studies and action research, accompanied by pilot programmes, are recommended as deliberate strategies for HSS.


Transport of bile acids in hepatic and non-hepatic tissues.
Bile acids are steroidal amphipathic molecules derived from the catabolism of cholesterol. They modulate bile flow and lipid secretion, are essential for the absorption of dietary fats and vitamins, and have been implicated in the regulation of all the key enzymes involved in cholesterol homeostasis. Bile acids recirculate through the liver, bile ducts, small intestine and portal vein to form an enterohepatic circuit. They exist as anions at physiological pH and, consequently, require a carrier for transport across the membranes of the enterohepatic tissues. Individual bile acid carriers have now been cloned from several species. Na+-dependent transporters that mediate uptake into hepatocytes and reabsorption from the intestine and biliary epithelium and an ATP-dependent transporter that pumps bile acids into bile comprise the classes of transporter that are specific for bile acids. In addition, at least four human and five rat genes that code for Na+-independent organic anion carriers with broad multi-substrate specificities that include bile acids have been discovered. Studies concerning the regulation of these carriers have permitted identification of molecular signals that dictate eventual changes in the uptake or excretion of bile acids, which in turn have profound physiological implications. This overview summarizes and compares all known bile acid transporters and highlights findings that have identified diseases linked to molecular defects in these carriers. Recent advances that have fostered a more complete appreciation for the elaborate disposition of bile acids in humans are emphasized.


Stemphylium Leaf Spot of Parsley in California Caused by Stemphylium vesicarium.
From 2009 through 2011, a previously undescribed disease occurred on commercial parsley in coastal (Ventura County) California. Symptoms of the disease consisted of circular to oval, tan to brown leaf spots and resulted in loss of crop quality and, hence, reduced yields. A fungus was consistently isolated from symptomatic parsley. Morphological and molecular data identified the fungus as Stemphylium vesicarium. When inoculated onto parsley leaves, the isolates caused symptoms that were identical to those seen in the field; the same fungus was recovered from test plants, thus completing Koch's postulates. Additional inoculation experiments demonstrated that 10 of 11 tested flat leaf and curly parsley cultivars were susceptible. The parsley isolates also caused small leaf spots on other Apiaceae family plants (carrot and celery) but not on leek, onion, spinach, and tomato. Isolates caused brown lesions to form when inoculated onto pear fruit but only when the fruit tissue was wounded. Using a freeze-blotter seedborne pathogen assay, parsley seed was found to have a low incidence (0.25%) of S. vesicarium. When inoculated onto parsley leaves, three of four isolates from seed caused the same leaf spot disease. This is the first documentation of a foliar parsley disease caused by S. vesicarium. The occurrence of S. vesicarium on parsley seed indicates that infested seed may be one source of initial inoculum. Based on the negative results in the host range experiments, it appears that this parsley pathogen differs from the S. vesicarium that causes disease on leek, garlic, onion, and pear fruit.


The potential use of the enhanced nonlinear properties of gold nanospheres in photothermal cancer therapy.
Laser photothermal therapy (PTT) is practiced at the moment using short laser pulses. The use of plasmonic nanoparticles as contrast agents can decrease the laser energy by using the optical property of the nanoparticles and improve the tumor selectivity by the molecular probes on the particle surface. In this study, we aim at selective and efficient PTT by exploiting the nonlinear optical properties of aggregated spherical gold nanoparticles conjugated to anti-epidermal growth factor receptor (anti-EGFR) antibodies using short NIR laser pulses. Spherical gold nanoparticles are synthesized and conjugated to anti-EGFR antibodies to specifically target HSC oral cancer cells. The nanoparticles are characterized by micro-absorption spectra and dark field light scattering imaging. Photothermal destructions of control and nanoparticle treated cancer cells are carried out with a femtosecond Ti:Sapphire laser at 800 nm with a pulse duration of 100 femtoseconds and repetition rate of 1 kHz. The laser power threshold for the photothermal destruction of cells after the nanoparticle treatment is found to be 20 times lower than that required to destroy the cells in the normal PTT, that is, without nanoparticles. The number of destroyed cells is quadratically dependent on the laser power. The number of dead cells shows a nonlinear dependence on the concentration of gold nanoparticles that are specifically targeted to cancer cells. The energy threshold and selectivity of PTT can greatly benefit from the use of the plasmonic enhanced nonlinear optical processes of spherical gold nanoparticles conjugated to anti-EGFR antibodies. The quadratic dependence of the photothermal efficiency on the pulsed NIR laser power indicates a second harmonic generation or a two photon absorption process. The observed nonlinear dependence on the gold nanoparticle concentration suggests that aggregated nanospheres are responsible for the observed enhanced photothermal destruction of the cells.


Alteration of the nucleosomal DNA path in the crystal structure of a human nucleosome core particle.
Gene expression in eukaryotes depends upon positioning, mobility and packaging of nucleosomes; thus, we need the detailed information of the human nucleosome core particle (NCP) structure, which could clarify chromatin properties. Here, we report the 2.5 A crystal structure of a human NCP. The overall structure is similar to those of other NCPs reported previously. However, the DNA path of human NCP is remarkably different from that taken within other NCPs with an identical DNA sequence. A comparison of the structural parameters between human and Xenopus laevis DNA reveals that the DNA path of human NCP consecutively shifts by 1 bp in the regions of superhelix axis location -5.0 to -2.0 and 5.0 to 7.0. This alteration of the human DNA path is caused predominantly by tight DNA-DNA contacts within the crystal. It is also likely that the conformational change in the human H2B tail induces the local alteration of the DNA path. In human NCP, the region with the altered DNA path lacks Mn2+ ions and the B-factors of the DNA phosphate groups are substantially high. Therefore, in contrast to the histone octamer, the nucleosomal DNA is sufficiently flexible and mobile and can undergo drastic conformational changes, depending upon the environment.


Proliferating cell nuclear antigen promotes misincorporation catalyzed by calf thymus DNA polymerase delta.
A proliferating cell nuclear antigen (PCNA)-dependent complex, detectable after nondenaturing polyacrylamide gel electrophoresis, is formed between calf thymus DNA polymerase delta (pol delta) and synthetic oligonucleotide template-primers containing a mispaired nucleotide at the 3'-terminal position of the primer. This complex is indistinguishable in composition from that formed with a fully base paired template-primer. Extension of a mispaired primer terminus is a component of DNA polymerase fidelity. The fidelity of pol delta on synthetic oligonucleotide template-primers was compared with and without its specific processivity factor, PCNA. In the absence of PCNA, pol delta misincorporates less than one nucleotide for every 100,000 nucleotides incorporated correctly. Addition of PCNA to reactions reduces fidelity by at least 27-fold. PCNA also confers upon pol delta, the ability to incorporate (and/or not excise) the dTTP analog, 2'-deoxythymidine-5'-O-(alpha-phosphonomethyl)-beta, gamma-diphosphate. A model is proposed whereby the increased stability (decreased off-rate) of the pol delta.template-primer complex in the presence of PCNA facilitates unfavorable events catalyzed by pol delta. This model suggests an explicit mechanistic requirement for the intrinsic 3'-5'-exonuclease of pol delta.


Neurofilament light chain is a promising serum biomarker in spinocerebellar ataxia type 3.
Spinocerebellar ataxia type 3 (SCA3) is the most common subtype of autosomal dominantly inherited spinocerebellar ataxias (SCAs). No validated blood biomarker is available to assess either disease progression or therapeutic response. Neurofilament light chain (NfL) was recently proposed as a serum biomarker for many neurodegenerative disorders. The present study investigated whether NfL was a promising serum biomarker for SCA3. Seventeen SCA3 patients and 9 controls were enrolled in cohort A, and 116 SCA3 individuals (preclinical and patients) and 91 controls were recruited as cohort B. We assessed whether serum NfL correlated with cerebrospinal fluid (CSF) NfL in cohort A and correlations between serum NfL levels and clinical features and brain volumes were determined in cohort B. The single-molecule array method was used to measure serum NfL levels. Disease severity was determined using the scale for the assessment and rating of ataxia (SARA) and the international cooperative ataxia rating scale (ICARS). Cerebellar and brainstem volumes were assessed using MRI neuroimaging measurements. Serum/CSF NfL levels in cohort A were elevated in SCA3 patients, and serum and CSF NfL exhibited a significant positive correlation (r = 0.9179, p < 0.0001). Levels of serum NfL in cohort B were significantly higher in preclinical SCA3 (15.03 ± 7.49 vs 6.88 ± 2.72 pg/ mL, p < 0.0001) and manifest SCA3 subjects (37.56 ± 13.47 vs 9.07 ± 6.02 pg/ mL, p < 0.0001) compared to those in controls. Serum NfL concentrations increased from early disease stage to the next stage. Levels of serum NfL in ATXN3 mutation carriers were positively associated with SARA (r = 0.5458, p < 0.0001) and ICARS scores (r = 0.5522, p < 0.0001). Significant negative associations with cerebellar volumes (r = - 0.4217, p = 0.0003) and brainstem volumes (r = - 0.4263, p = 0.0003) were observed. All changes remained significant after adjustment for age and CAG repeat. Levels of serum NfL were significantly elevated in SCA3 individuals and correlated with disease severity. Serum NfL is a promising serum biomarker of disease onset and progression, and a potential candidate biomarker of treatment response in SCA3.


Rer1p maintains ciliary length and signaling by regulating γ-secretase activity and Foxj1a levels.
Cilia project from the surface of most vertebrate cells and are important for several physiological and developmental processes. Ciliary defects are linked to a variety of human diseases, named ciliopathies, underscoring the importance of understanding signaling pathways involved in cilia formation and maintenance. In this paper, we identified Rer1p as the first endoplasmic reticulum/cis-Golgi-localized membrane protein involved in ciliogenesis. Rer1p, a protein quality control receptor, was highly expressed in zebrafish ciliated organs and regulated ciliary structure and function. Both in zebrafish and mammalian cells, loss of Rer1p resulted in the shortening of cilium and impairment of its motile or sensory function, which was reflected by hearing, vision, and left-right asymmetry defects as well as decreased Hedgehog signaling. We further demonstrate that Rer1p depletion reduced ciliary length and function by increasing γ-secretase complex assembly and activity and, consequently, enhancing Notch signaling as well as reducing Foxj1a expression.


Development of Allergic Conjunctivitis Induced by House Dust Mite Extract From Dermatophagoides pteronyssinus.
The purpose of this study was to develop a murine model of allergic conjunctivitis induced by house dust mite (HDM) extract from Dermatophagoides pteronyssinus, a major allergen in humans. Forty BALB/c mice were divided into five groups, immunized with placebo, ovalbumin (10 μg), or HDM extract following a schedule. Twenty minutes after topical challenge, mice were examined clinically. Material collected from mice was used for measuring total and specific IgE, antigen-specific lymphocyte proliferation, and supernatant cytokine levels and for conjunctival histopathology and flow cytometric analysis of conjunctival cells. This murine model showed similar clinical signs and laboratory findings to human allergy and the ovalbumin-induced allergic conjunctivitis model. Total IgE levels and conjunctival infiltration of mast cells and eosinophils in immunized mice were significantly higher than in the control group. Cervical lymphocyte proliferation was increased in antigen-stimulated cultures in immunized mice, concomitant with significantly higher IL-4 and IL-5 levels in the culture supernatant. The proportion of conjunctival CD4+ T cells expressing the ST2 receptor was increased, and conjunctival CD4+ST2+ T cells exhibited an increase in intracellular IL-5. House dust mite extract successfully induced allergic conjunctivitis in BALB/c mice. Ten micrograms of HDM extract was the optimal dose for systemic immunization in this model. This murine model is suitable for further studies on HDM-induced allergic conjunctivitis, and the data show that conjunctival CD4+ T cells expressing ST2 may play an important role in IL-5 secretion, recruiting eosinophils into conjunctiva on ocular allergen challenge.


Embryonic but not postnatal reexpression of hepatocyte nuclear factor 1alpha (HNF1alpha) can reactivate the silent phenylalanine hydroxylase gene in HNF1alpha-deficient hepatocytes.
The failure to transcribe the phenylalanine hydroxylase (PAH) gene in the liver of hepatocyte nuclear factor 1alpha (HNF1alpha)-deficient mice correlated with DNA hypermethylation and the presence of an inactive chromatin structure (M. Pontoglio, D. M. Faust, A. Doyen, M. Yaniv, and M. C. Weiss, Mol. Cell. Biol. 17:4948-4956, 1997). To evaluate the precise role played by HNF1alpha, DNA methylation, or histone acetylation in PAH gene silencing, we examined conditions that could restore PAH gene expression in HNF1alpha-deficient hepatocytes. We show that reactivation of PAH transcription can be achieved by reexpression of HNF1alpha in embryonic (i.e., embryonic day 12.5 [e12.5] to e13.5) hepatocytes but not in fetal (e17.5), newborn, and adult HNF1alpha-deficient hepatocytes. This defines a temporal competence window during which HNF1alpha can act to (re)program PAH gene transcription. We also show that PAH gene silencing can be partially relieved in HNF1alpha-deficient hepatocytes by treatment with the demethylating agent 5-azacytidine, even in the absence of HNF1alpha. Treatment using 5-azacytidine combined with trichostatin, a histone deacetylase inhibitor, resulted in a synergistic reactivation of the silenced PAH gene in adult hepatocytes, but this activity was not further increased by HNF1alpha reexpression. These results suggest that the HNF1alpha homeoprotein is involved in stage-specific developmental control of the methylation state and chromatin remodeling of the PAH gene.


The development of therapeutic monoclonal antibodies: overview of the nonclinical safety assessment.
Monoclonal antibodies (mAbs) represent a class of biotechnology-derived therapeutics for use in the treatment of various disease indications such as oncology, autoimmune, cardiovascular, and metabolic disorders. Monoclonal antibodies are immunoglobulin (Ig) proteins engineered to bind to specific antigens with high specificity. The concepts reviewed in this paper include 1) the regulatory procedures and guidelines that apply to mAbs, 2) the types of toxicology studies applicable to mAbs, and 3) the scientific challenges, such as the selection of a relevant animal species and the development of anti-drug antibodies, that can arise due to the unique properties of mAbs.


Decontamination and Management of Contaminated Hair following a CBRN or HazMat Incident.
This in vitro study evaluated the "triple protocol" of dry decontamination, the ladder pipe system (a method for gross decontamination), and technical decontamination for the decontamination of hair following chemical contamination. First, we assessed the efficacy of the 3 protocols, alone or in combination, on excised porcine skin and human hair contaminated with either methyl salicylate (MS), phorate (PHR), sodium fluoroacetate (SFA), or potassium cyanide (KCN). A second experiment investigated the residual hair contamination following decontamination with the triple protocol at different intervals postexposure. In a third experiment, hair decontaminated after exposure to MS or PHR was evaluated for off-gassing. Though skin decontamination was highly effective, a substantial proportion (20%-40%) of the lipophilic compounds (MS and PHR) remained within the hair. The more water-soluble contaminants (SFA and KCN) tended to form much smaller reservoirs within the hair. Interestingly, substantial off-gassing of MS, a medium volatility chemical, was detectable from triple-decontaminated hair up to 5 days postexposure. Overall, the decontamination strategies investigated were effective for the decontamination of skin, but less so for hair. These findings highlight the importance of contaminated hair serving as a source of potential secondary contamination by contact or inhalation. Therefore, consideration should be given to the removal of contaminated hair following exposure to toxic chemicals.


Changes in blood glucose and plasma insulin after intravenous galactose in human injury.
1. The increase in blood glucose after intravenous galactose was measured at 6--8 h and 14 days after hip replacement (14 patients) and accidental injury (14 patients). 2. After hip replacement there was a greater rise of glucose after galactose on the day of the operation than on recovery, despite basal hyperglycaemia. This earlier period was also associated with inappropriately low insulin concentrations for the prevailing glucose concentration, and hyperketonaemia (13 out 14 patients). 3. After accidental injury patients with initial hyperketonaemia (nine out of 14) also had a greater rise of blood glucose after galactose than on recovery and had relative insulin deficiency; in contrast those who were initially normoketonaemic (five out of 14) showed a rise in glucose comparable with that after recovery and basal insulin concentrations more appropriate to the existing glucose concentration. 4. It is concluded that in most subjects after injury hepatic glucose release after galactose administration is not suppressed despite hyperglycaemia.


Hydrophobic properties of polytetrafluoroethylene thin films fabricated at various catalyzer temperatures through catalytic chemical vapor deposition using a tungsten catalyzer.
Using the catalytic chemical vapor deposition (Cat-CVD) method, polytetrafluoroethylene (PTFE) thin films were fabricated on Si(100) substrates at various catalyzer temperatures, using a tungsten catalyzer, and Fourier transform infrared (FTIR) spectroscopy and X-ray photoemission spectroscopy (XPS) were used to confirm the fabrication of the films. An atomic-force microscope (AFM) and a scanning electron microscope (SEM) were employed to study the correlation between the wettability and surface morphology of the samples. It was found that the wettability of the PTFE thin films fabricated via Cat-CVD is strongly correlated with the sizes of the film surfaces' nanoprotrusions, and that superhydrophobic PTFE thin-film surfaces can be easily achieved by controlling the sizes of the nanoprotrusions through the catalyzer temperature. The comparison of the wettability values and surface morphologies of the films confirmed that nanoscale surface roughness enhances the hydrophobic properties of PTFE thin films. Further, the detailed analysis of the films' surface morphologies from their AFM images with the use of the Wenzel and Cassie models confirmed that the nanoscale surface roughness enhanced the hydrophobic property of the PTFE films. Further, the variations of the wettability of the PTFE thin films prepared via Cat-CVD are well explained by the Cassie model. It seems that the increase in the trapping air and the reduction of the liquid-solid contact area are responsible for the superhydrophobicity of the PTFE thin films prepared via Cat-CVD.


The influence of diet on unconjugated hyperbilirubinaemia in the Gunn rat.
1. The influence of dietary carbohydrate, protein and lipid on the unconjugated hyperbilirubinaemia of Gunn rats has been studied. 2. In the first set of experiments, after a control period without restraint, the animals were restrained for 72 h and were either starved or received isocaloric amounts of glucose, protein hydrolysate or soya bean oil through a gastrotomy cannula. 3. A second study was performed with groups of unrestrained animals which were fed with diets supplemented with carbohydrate, protein and lipid for 7 days after a control period. 4. Both these studies showed that diets composed predominantly of carbohydrate or protein doubled the plasma bilirubin concentration. Starch, glucose and fructose produced similar effects. 5. A high-lipid diet in the absence of carbohydrate caused a slight reduction plasma bilirubin concentration. This effect was observed with both soya bean oil and pork fat. 6. It has not been established whether the increase in hyperbilirubinaemia resulted from the high-carbohydrate content of the diet or the absence of lipid.


Virtual Embodiment of White People in a Black Virtual Body Leads to a Sustained Reduction in Their Implicit Racial Bias.
Virtual reality can be used to visually substitute a person's body by a life-sized virtual one. Such embodiment results in a perceptual illusion of body ownership over the virtual body (VB). Previous research has shown that the form of the VB can influence implicit attitudes. In particular, embodying White people in a Black virtual body is associated with an immediate decrease in their implicit racial bias against Black people. We tested whether the reduction in implicit bias lasts for at least 1 week and whether it is enhanced by multiple exposures. Two experiments were carried out with a total of 90 female participants where the virtual body was either Black or White. Participants were required to follow a virtual Tai Chi teacher who was either Asian or European Caucasian. Each participant had 1, 2, or 3 exposures separated by days. Implicit racial bias was measured 1 week before their first exposure and 1 week after their last. The results show that implicit bias decreased more for those with the Black virtual body than the White. There was also some evidence of a general decrease in bias independently of body type for which possible explanations are put forward.


BiopSee® - transperineal stereotactic navigated prostate biopsy.
In the recent years, prostate cancer was the most commonly diagnosed cancer in men. Currently secure diagnosis confirmation is done by a transrectal biopsy and following histopathological examination. Conventional transrectal biopsy success rates are rather low with ca. 30% detection upon the first and ca 20% after re-biopsy. The paper presents a novel system for stereotactic navigated prostate biopsy. The approach results into higher accuracy, reproducibility and unrestricted and effective access to all prostate regions. Custom designed ultrasound, new template design and integrated 2-axes stepper allows superior 2D and 3D prostate imaging quality and precise needle navigation. DICOM functionality and image fusion enable to import pre-operative datasets (e.g. multiparametric MRI, targets etc.) and overlay all available radiological information into the biopsy planning and guiding procedure. The biopsy needle insertion itself is performed under augmented reality ultrasound guidance. Each procedure step is automatically documented in order to provide quality assurance and permit data re-usage for the further treatment. First clinical results indicates success rates of ca. 70% by first biopsies by our approach.


[SARS-CoV-2 mRNA-based vaccine in hemodialysis patients: a single center-experience].
Hemodialysis patients have an increased risk of severe complications when infected with SARS-CoV-2. The introduction of the SARS-CoV-2 vaccine represented an important progress in limiting severe forms of the disease. The focus of our study is the detection of the antibody titer in chronic hemodialysis patients vaccinated with the mRNA vaccine BNT162b2 (Comirnaty, Pfizer-BioNTech). The antibody titers were measured in 57 hemodialysis patients, vaccinated with 3 doses according to ministerial criteria, by ElectroChemiLuminescence ImmunoAssay (ECLIA). The response was defined as an antibody titer above the dosable level > 0,8 UI/ml. A good antibody response was defined as titer > 250 UI/ml. Infections with SARS-CoV-2 and adverse effects to the vaccine were recorded. Our study showed in 93% of the hemodialysis patients a dosable antibody response after the second dose of the vaccine. After the third dose of the vaccine, 100% of the hemodialysis patients reached a dosable antibody titer. The vaccine proved to be safe, no serious adverse events were observed. After the third dose, SARS-CoV-2 infections were still observed, but with reduced severity. A vaccination course against SARS-CoV-2 infection with three doses of BNT162b2 in the dialysis patient is associated with a good immune response and protects against severe infections.


Influence of carotid baroreceptors on vascular responses to carotid chemoreceptor stimulation in the dog.
The modification by the carotid baroreceptors of the vascular responses to chemoreceptor stimulation was studied in anesthetized, artificially ventilated, vagotomized dogs. The carotid bifurcations were vascularly isolated and perfused with blood at constant pressures of 134, 215, and 51 mm Hg to cause intermediate, maximal, and minimal inhibition, respectively, of the vasomotor center. At each pressure, stimulation of the carotid chemoreceptors was achieved by perfusion with hypoxic hypercapnic blood. With intermediate inhibition, the chemoreceptor stimulation increased the aortic pressure by 50% and decreased the hind-limb and kidney blood flow (perfusion at constant pressure) by 59% and 19%, respectively. At carotid sinus pressures of 215 and 51 mm Hg, the effects of chemoreceptor stimulation were absent or markedly attenuated. With intermediate sinus pressure, chemoreceptor stimulation decreased the perfusion pressure of the saphenous vein by 27% (perfusion at constant flow). When the sinus pressure was increased to 215 mm Hg, the tone of the vein did not change, but chemoreceptor stimulation was without effect. The present study indicates a central interaction (which may be presynaptic) between the chemoreceptor and baroreceptor inputs such that the vascular responses to chemoreceptor stimulation are inhibited when the carotid sinus activity is maximal or minimal.


Surveillance for human West Nile virus disease - United States, 1999-2008.
West Nile virus (WNV) is an arthropod-borne virus (arbovirus) in the family Flaviviridae and is the leading cause of arboviral disease in the United States. An estimated 80% of WNV infections are asymptomatic. Most symptomatic persons develop an acute systemic febrile illness that often includes headache, myalgia, arthralgia, rash, or gastrointestinal symptoms. Less than 1% of infected persons develop neuroinvasive disease, which typically presents as encephalitis, meningitis, or acute flaccid paralysis. 1999-2008. WNV disease is a nationally notifiable disease with standardized case definitions. State and metropolitan heath departments report cases to CDC through ArboNET, an electronic passive surveillance system. Variables collected include patient age, sex, race, county and state of residence, date of illness onset, clinical syndrome, and outcome of illness. During 1999-2008, a total of 28,961 confirmed and probable cases of WNV disease, including 11,822 (41%) WNV neuroinvasive disease cases, were reported to CDC from 47 states and the District of Columbia. No cases were reported from Alaska, Hawaii, Maine, or any U.S. territories. A total of 93% of all WNV patients had illness onset during July-September. The national incidence of WNV neuroinvasive disease peaked in 2002 (1.02 cases per 100,000 population) and was stable during 2004-2007 (mean annual incidence: 0.44; range: 0.39-0.50). In 2008, the incidence was 0.23 per 100,000 population, compared with 0.41 in 2007 and 0.50 in 2006. During 1999-2008, the highest incidence of neuroinvasive disease occurred in West North Central and Mountain states. Neuroinvasive disease incidence increased with increasing age, with the highest incidence (1.35 cases per 100,000 population) occurring among persons aged >or=70 years. The hospitalization rate and case-fatality ratio increased with increasing age among persons with neuroinvasive disease. The stability in reported incidence of neuroinvasive disease during 2004-2007 might represent an endemic level of WNV transmission. Whether the incidence reported in 2008 represents a decrease that will continue is unknown; variations in vectors, avian amplifying hosts, human activity, and environmental factors make predicting future WNV transmission levels difficult. Surveillance of WNV disease is important for detecting and monitoring seasonal epidemics and targeting prevention and control activities. Public health education programs should focus on older persons, who are at increased risk for neurologic disease and poor clinical outcomes. In the absence of an effective human vaccine, WNV disease prevention depends on community-level mosquito control and household and personal protection measures.


Glaciers. Attribution of global glacier mass loss to anthropogenic and natural causes.
The ongoing global glacier retreat is affecting human societies by causing sea-level rise, changing seasonal water availability, and increasing geohazards. Melting glaciers are an icon of anthropogenic climate change. However, glacier response times are typically decades or longer, which implies that the present-day glacier retreat is a mixed response to past and current natural climate variability and current anthropogenic forcing. Here we show that only 25 ± 35% of the global glacier mass loss during the period from 1851 to 2010 is attributable to anthropogenic causes. Nevertheless, the anthropogenic signal is detectable with high confidence in glacier mass balance observations during 1991 to 2010, and the anthropogenic fraction of global glacier mass loss during that period has increased to 69 ± 24%.


Developmentally regulated activation of apoptosis early in Xenopus gastrulation results in cyclin A degradation during interphase of the cell cycle.
Previous work identified a developmental timer that controls the stability of cyclin A protein in interphase-arrested Xenopus embryos. It was shown that cyclins A1 and A2 abruptly become unstable in hydroxyurea-treated embryos at the time that untreated embryos are beginning gastrulation (early gastrulation transition; EGT). We have demonstrated here that cyclins A1 and A2 are degraded at the equivalent of the EGT by the ICE-like caspases that are responsible for programmed cell death or apoptosis. Analysis of embryos treated with hydroxyurea or cycloheximide showed widespread cellular apoptosis coincident with cyclin A cleavage. Our data further indicate that the apoptotic pathway is present in Xenopus embryos prior to the EGT; however, it is maintained in an inactive state in early cleaving embryos by maternally encoded inhibitors. Characterization of the timing of the activation of apoptosis implicates the initiation of zygotic transcription at the mid-blastula transition (MBT) in the suppression of apoptosis in normal embryos. The decreased biosynthetic capacity of embryos treated with hydroxyurea or cycloheximide most likely interferes with the ability to maintain sufficient levels of apoptotic inhibitors and results in widespread apoptosis. Our results suggest a scenario whereby the apoptotic pathway is suppressed in the early cleaving embryo by maternally contributed inhibitors. Degradation at the EGT of maternal RNAs encoding these inhibitors is compensated for by new zygotic transcription beginning at the MBT. This indicates that the interval between the MBT and the EGT represents a critical developmental period during which the regulation of embryonic cellular processes is transferred from maternal to zygotic control.


The palaeoenvironmental impact of prehistoric settlement and proto-historic urbanism: tracing the emergence of the Oppidum of Corent, Auvergne, France.
Early human societies and their interactions with the natural world have been extensively explored in palaeoenvironmental studies across Central and Western Europe. Yet, despite an extensive body of scholarship, there is little consideration of the environmental impacts of proto-historic urbanisation. Typically palaeoenvironmental studies of Bronze and Iron Age societies discuss human impact in terms of woodland clearance, landscape openness and evidence for agriculture. Although these features are clearly key indicators of human settlement, and characterise Neolithic and early to Middle Bronze Age impacts at Corent, they do not appear to represent defining features of a protohistoric urban environment. The Late Iron Age Gallic Oppidum of Corent is remarkable for the paucity of evidence for agriculture and strong representation of apophytes associated with disturbance. Increased floristic diversity - a phenomenon also observed in more recent urban environments - was also noted. The same, although somewhat more pronounced, patterns are noted for the Late Bronze Age and hint at the possibility of a nascent urban area. High percentages of pollen from non-native trees such as Platanus, Castanea and Juglans in the late Bronze Age and Gallic period also suggest trade and cultural exchange, notably with the Mediterranean world. Indeed, these findings question the validity of applying Castanea and Juglans as absolute chronological markers of Romanisation. These results clearly indicate the value of local-scale palaeoecological studies and their potential for tracing the phases in the emergence of a proto-historic urban environment.


Early treatment of subarachnoid hemorrhage after preventing rerupture of an aneurysm.
Twelve patients with Hunt and Hess neurological Grades III to V underwent thrombosis of aneurysms using cellulose acetate polymer within 23 hours of aneurysm rupture. On computerized tomography (CT), nine of these patients had difuse or localized thick subarachnoid blood clots, two had diffuse thin clots and one had intraventricular clots. Immediately after thrombosis, all patients were administered tissue plasminogen activator (TPA) through spinal or ventricular catheters. The pressure of the lumbar cerebrospinal fluid was maintained at 100 to 150 mm H2O. The TPA was given as multiple injections of 2 mg on Day 0 and 1 to 2 mg on the following 1 to 2 days. In two patients the second injection of TPA was not given because of severe brain damage resulting from the initial subarachnoid hemorrhage. Ten patients showed complete clearance of the cisternal clot on CT within 72 hours after thrombosis. Seven partially thrombosed aneurysms and five multiple aneurysms were clipped during delayed surgery. Only one patient experienced mild vasospasm as shown on the follow-up angiogram. Eight patients improved clinically and had a good recovery, two had severe disability, and two died. Urgent thrombosis of a ruptured aneurysm followed by immediate postthrombotic administration of TPA may be a safe and reasonable means of preventing vasospasm and improving patient outcome.


Molecular epidemiology of a fast-food restaurant-associated outbreak of Escherichia coli O157:H7 in Washington State.
We studied the molecular epidemiology of the recent fast-food restaurant chain-associated Escherichia coli O157:H7 outbreak in Washington State. Genomic DNAs prepared from strains isolated from 433 patients were probed with radiolabelled Shiga-like toxin (SLT) I and SLT II genes and bacteriophage lambda DNA and were subsequently analyzed for their restriction fragment length polymorphism (RFLP) patterns. The SLT RFLP and lambda RFLP profiles of an E. coli O157:H7 strain isolated from the incriminated beef and prototype patient were compared with those of the patient isolates for determination of the concordance between patterns. Of the 377 patients with primary and secondary cases of infection epidemiologically linked to the outbreak, isolates from 367 (97.3%) of the patients displayed SLT RFLP and lambda RFLP profiles identical to those of the outbreak strains. Isolates from 10 of the 377 (2.6%) patients possessed SLT RFLP and lambda RFLP profiles different from those of the outbreak strains, and the patients from whom those isolates were obtained were subsequently characterized as having non-outbreak-related infections. The E. coli O157:H7 strains isolated from 31 of 44 (70.4%) patients who were epidemiologically excluded from the outbreak were linked to the outbreak by RFLP typing. Our results indicate that SLT RFLP and lambda RFLP analyses are stable and sensitive methods, and when they are used in conjunction with an epidemiological investigation they could result in an earlier recognition of outbreaks and their sources, hence prompting measures to prevent the continued transmission of E. coli O157:H7.


Streptococcal sagA activates a proinflammatory response in mast cells by a sublytic mechanism.
Mast cells are implicated in the innate proinflammatory immune defence against bacterial insult, but the mechanisms through which mast cells respond to bacterial encounter are poorly defined. Here, we addressed this issue and show that mast cells respond vividly to wild type Streptococcus equi by up-regulating a panel of proinflammatory genes and by secreting proinflammatory cytokines. However, this response was completely abrogated when the bacteria lacked expression of sagA, whereas the lack of a range of other potential virulence genes (seeH, seeI, seeL, seeM, hasA, seM, aroB, pyrC, and recA) had no effect on the amplitude of the mast cell responses. The sagA gene encodes streptolysin S, a lytic toxin, and we next showed that the wild type strain but not a sagA-deficient mutant induced lysis of mast cells. To investigate whether host cell membrane perturbation per se could play a role in the activation of the proinflammatory response, we evaluated the effects of detergent- and pneumolysin-dependent lysis on mast cells. Indeed, exposure of mast cells to sublytic concentrations of all these agents resulted in cytokine responses of similar amplitudes as those caused by wild type streptococci. This suggests that sublytic membrane perturbation is sufficient to trigger full-blown proinflammatory signalling in mast cells. Subsequent analysis showed that the p38 and Erk1/2 signalling pathways had central roles in the proinflammatory response of mast cells challenged by either sagA-expressing streptococci or detergent. Altogether, these findings suggest that sagA-dependent mast cell membrane perturbation is a mechanism capable of activating the innate immune response upon bacterial challenge.


Relations between subclinical disease markers and type 2 diabetes, metabolic syndrome, and incident cardiovascular disease: the Jackson Heart Study.
The presence of subclinical disease measures has been directly associated with the development of cardiovascular disease (CVD) in whites. African Americans (AAs) in the U.S. are at higher risk of CVD compared with non-Hispanic whites; however, data on the prevalence of subclinical disease measures in AAs and their association to CVD remain unclear and may explain the higher CVD risk in this group. We evaluated 4,416 participants attending the first examination of the Jackson Heart Study (mean age 54 years; 64% women) with available subclinical disease measures. There were 1,155 participants (26%) with subclinical disease, defined as the presence of one or more of the following: peripheral arterial disease, left ventricular hypertrophy, microalbuminuria, high coronary artery calcium (CAC) score, and low left ventricular ejection fraction. In cross-sectional analyses using multivariable-adjusted logistic regression, participants with metabolic syndrome (MetS) or diabetes (DM) had higher odds of subclinical disease compared with those without MetS and DM (odds ratios 1.55 [95% CI 1.30-1.85] and 2.86 [95% CI 2.32-3.53], respectively). Furthermore, the presence of a high CAC score and left ventricular hypertrophy were directly associated with the incidence of CVD (265 events) in multivariable-adjusted Cox proportional hazards regression models (P < 0.05). In prospective analyses, having MetS or DM significantly increased the hazard of incident CVD, independent of the presence of subclinical disease (P < 0.001). In our community-based sample of AAs, we observed a moderately high prevalence of subclinical disease, which in turn translated into a greater risk of CVD, especially in people with MetS and DM.


Imaging the photodissociation dynamics of the methyl radical from the 3s and 3pz Rydberg states.
The photodissociation dynamics of the methyl radical from the 3s and 3pz Rydberg states have been studied using the velocity map and slice ion imaging in combination with pump-probe nanosecond laser pulses. The reported translational energy and angular distributions of the H((2)S) photofragment detected by (2+1) REMPI highlight different dissociation mechanisms for the 3s and 3pz Rydberg states. A narrow peak in the translational energy distribution and an anisotropic angular distribution characterize the fast 3s photodissociation, while for the 3pz state Boltzmann-type translational energy and isotropic angular distributions are found. High level ab initio calculations have been performed in order to elucidate the photodissociation mechanisms from the two Rydberg states and to rationalize the experimental results. The calculated potential energy curves highlight a typical predissociation mechanism for the 3s state, characterized by the coupling between the 3s Rydberg state and a valence repulsive state. On the other hand, the photodissociation on the 3pz state is initiated by a predissociation process due to the coupling between the 3pz Rydberg state and a valence repulsive state and constrained, later on, by two conical intersections that allow the system to relax to lower electronic states. Such a mechanism opens up different reaction pathways leading to CH2 photofragments in different electronic states and inducing a transfer of energy between translational and internal modes.


Theoretical realization of robust broadband transparency in ultrathin seamless nanostructures by dual blackbodies for near infrared light.
We propose a counter-intuitive mechanism of constructing an ultrathin broadband transparent device with two perfect blackbodies. By introducing hybridization of plasmon modes, resonant modes with different symmetries coexist in this system. A broadband transmission spectrum in the near infrared regime is achieved through controlling their coupling strengths, which is governed by the thickness of high refractive index layer. Meanwhile, the transparency bandwidth is found to be tunable in a large range by varying the geometric dimension. More significantly, from the point view of applications, the proposed method of achieving broadband transparency can perfectly tolerate the misalignment and asymmetry of periodic nanoparticles on the top and bottom, which is empowered by the unique dual of coupling-in and coupling-out processes within the pair of blackbodies. Moreover, roughness has little influence on its transmission performance. According to the coupled mode theory, the distinguished transmittance performance is physically interpreted by the radiative decay rate of the entire system. In addition to the feature of uniquely robust broadband transparency, such a ultrathin seamless nanostructure (in the presence of a uniform silver layer) also provides polarization-independent and angle-independent operations. Therefore, it may power up a wide spectrum of exciting applications in thin film protection, touch screen techniques, absorber-emitter transformation, etc.


Process evaluation of appreciative inquiry to translate pain management evidence into pediatric nursing practice.
Appreciative inquiry (AI) is an innovative knowledge translation (KT) intervention that is compatible with the Promoting Action on Research in Health Services (PARiHS) framework. This study explored the innovative use of AI as a theoretically based KT intervention applied to a clinical issue in an inpatient pediatric care setting. The implementation of AI was explored in terms of its acceptability, fidelity, and feasibility as a KT intervention in pain management. A mixed-methods case study design was used. The case was a surgical unit in a pediatric academic-affiliated hospital. The sample consisted of nurses in leadership positions and staff nurses interested in the study. Data on the AI intervention implementation were collected by digitally recording the AI sessions, maintaining logs, and conducting individual semistructured interviews. Data were analysed using qualitative and quantitative content analyses and descriptive statistics. Findings were triangulated in the discussion. Three nurse leaders and nine staff members participated in the study. Participants were generally satisfied with the intervention, which consisted of four 3-hour, interactive AI sessions delivered over two weeks to promote change based on positive examples of pain management in the unit and staff implementation of an action plan. The AI sessions were delivered with high fidelity and 11 of 12 participants attended all four sessions, where they developed an action plan to enhance evidence-based pain assessment documentation. Participants labeled AI a 'refreshing approach to change' because it was positive, democratic, and built on existing practices. Several barriers affected their implementation of the action plan, including a context of change overload, logistics, busyness, and a lack of organised follow-up. Results of this case study supported the acceptability, fidelity, and feasibility of AI as a KT intervention in pain management. The AI intervention requires minor refinements (e.g., incorporating continued follow-up meetings) to enhance its clinical utility and sustainability. The implementation process and effectiveness of the modified AI intervention require evaluation in a larger multisite study.


Brain injury markers in new-onset seizures in adults: A pilot study.
Biochemical markers of brain pathology could potentially contribute to diagnosis and prediction in epilepsy. We describe levels of five brain injury markers in adults with new-onset seizures, and assess group differences in patients with a single seizure, epilepsy, and poststroke epilepsy. In this prospective observational study, adults with new-onset seizures were recruited at Sahlgrenska University Hospital, Sweden, and concentrations of glial fibrillary acidic protein (GFAP), neurofilament light (NfL), microtubule-associated protein tau (tau), S100 calcium-binding protein (S100B), and neuron-specific enolase (NSE) were measured. Participants were categorized as epilepsy, poststroke epilepsy (PSE), or single seizure (no additional seizures). Patients were followed until a diagnosis of epilepsy or PSE, or for at least two years in single seizure cases. The cohort included 23 (37%) individuals with a single seizure, 24 (39%) with epilepsy, and 15 (24%) with PSE. The concentrations of S100B were higher in patients with epilepsy and PSE than in single seizures (p = 0.0023 and p = 0.0162, respectively). The concentrations of NfL were higher in patients with PSE than in single seizures (p=0.0027). After age-normalization, levels of S100B were higher in patients with epilepsy and levels of NfL were higher in patients with PSE (p = 0.0021 and p = 0.0180). Levels of S100B and NfL were higher in patients with epilepsy or PSE than patients with single seizures. Further studies are needed to investigate the biomarker potential of brain injury markers as predictors of epilepsy course or indicators of epileptogenesis.


Revision surgery due to magnet dislocation in cochlear implant patients: an emerging complication.
To analyze the cause and effect of magnet dislocation in cochlear implant (CI) recipients requiring magnet revision surgery for treatment. Retrospective study. Tertiary referral center. Case reports from 1,706 CI recipients consecutively implanted from January 2000 to December 2011 were reviewed. The number of cases requiring magnet revision surgery was assessed. Revision surgery involving magnet removal or replacement was indicated in 1.23% (21/1,706), of all CI recipients. Magnet dislocation occurring during magnetic resonance tomography (MRI), at 1.5 Tesla (T), with the magnet in place and with the application of compression bandaging around the head, was the main cause for revision surgery in 47.62% (10/21) of the affected cases. All 10 cases were implanted with Cochlear Nucleus cochlear implants. These events occurred, despite adherence to current recommendations of the manufacturer. The present study underlines that MRI examination is the main cause of magnet dislocation. The use of compressive bandaging when using 1.5-T MRI does not eliminate the risk of magnet dislocation. Additional cautionary measures are for required for conditional MRI. We recommend X-ray examination after MRI to determine magnet dislocation and avoid major complications in all cases reporting pain during or after MRI. Additional research regarding silicon magnet pocket design for added retention is needed. Effective communication of guidelines for precautionary measures during MRI examination in CI patients is mandatory for all clinicians involved. MRI in CI recipients should be indicated with caution.


Loss of intestinal GATA4 prevents diet-induced obesity and promotes insulin sensitivity in mice.
Transcriptional regulation of small intestinal gene expression controls plasma total cholesterol (TC) and triglyceride (TG) levels, which are major determinants of metabolic diseases. GATA4, a zinc finger domain transcription factor, is critical for jejunal identity, and intestinal GATA4 deficiency leads to a jejunoileal transition. Although intestinal GATA4 ablation is known to misregulate jejunal gene expression, its pathophysiological impact on various components of metabolic syndrome remains unknown. Here, we used intestine-specific GATA4 knockout (GATA4iKO) mice to dissect the contribution of GATA4 on obesity development. We challenged adult GATA4iKO mice and control littermates with a Western-type diet (WTD) for 20 wk. Our findings show that WTD-fed GATA4iKO mice are resistant to diet-induced obesity. Accordingly, plasma TG and TC levels are markedly decreased. Intestinal lipid absorption in GATA4iKO mice was strongly reduced, whereas luminal lipolysis was unaffected. GATA4iKO mice displayed a greater glucagon-like peptide-1 (GLP-1) release on normal chow and even after long-term challenge with WTD remained glucose sensitive. In summary, our findings show that the absence of intestinal GATA4 has a beneficial effect on decreasing intestinal lipid absorption causing resistance to hyperlipidemia and obesity. In addition, we show that increased GLP-1 release in GATA4iKO mice decreases the risk for development of insulin resistance.


Cold stress-induced modulation of cell immunity during acute Toxoplasma gondii infection in mice.
Infection with Toxoplasma gondii in the acute phase results in nonspecific suppression of immunologic function in mice and humans. The present study examined the effects of a physical stressor, i.e., cold stress (CS), on macrophage function (nitrite production, parasite survival) and splenic blastogenesis in the acute phase of murine T. gondii infection. In our stress paradigm, female BALB/c mice were placed in cold water (1 +/- 0.5 C), 5 min each day for 8 days. Nitrite production and parasite survival were measured in cultured peritoneal macrophages obtained from mice subjected to CS after in vivo activation with interferon-gamma/lipopolysaccharide (CS + ACT), and in vitro infection with T. gondii tachyzoites. Peritoneal macrophages from CS + ACT mice showed decreased nitrite production compared to control but activated cells (ACT). Spleen cell proliferation to in vitro stimulation with the mitogens concanavalin A (Con A) and anti-CD3, and Toxoplasma lysate antigen (TLA) was measured in splenocytes obtained from BALB/c mice during the acute phase of infection with T. gondii. Mice subjected to CS and infection (CS + INF) had maximum splenocyte proliferation on days 8 and 15 followed by a subsequent decline on day 28 postinoculation (PI). In contrast, infected mice not subjected to stress (INF) showed decreased splenocyte proliferation on days 8 and 15 followed by an increase on day 28 PI. The rate of mortality was decreased in the CS + INF compared to the INF group during acute infection. These results suggest that CS may alter the pathogenesis of T. gondii infection by modulating acute-phase responses, provoking a state of transient disequilibrium between the host and parasite.


An evaluation of acute toxicity of colloidal silver nanoparticles.
Tests for acute oral toxicity, eye irritation, corrosion and dermal toxicity of colloidal silver nanoparticles (AgNPs) were conducted in laboratory animals following OECD guidelines. Oral administration of AgNPs at a limited dose of 5,000 mg/kg produced neither mortality nor acute toxic signs throughout the observation period. Percentage of body weight gain of the mice showed no significant difference between control and treatment groups. In the hematological analysis, there was no significant difference between mice treated with AgNPs and controls. Blood chemistry analysis also showed no differences in any of the parameter examined. There was neither any gross lesion nor histopathological change observed in various organs. The results indicated that the LD(50) of colloidal AgNPs is greater than 5,000 mg/kg body weight. In acute eye irritation and corrosion study, no mortality and toxic signs were observed when various doses of colloidal AgNPs were instilled in guinea pig eyes during 72 hr observation period. However, the instillation of AgNPs at 5,000 ppm produced transient eye irritation during early 24 hr observation time. No any gross abnormality was noted in the skins of the guinea pigs exposed to various doses of colloidal AgNPs. In addition, no significant AgNPs exposure relating to dermal tissue changes was observed microscopically. In summary, these findings of all toxicity tests in this study suggest that colloidal AgNPs could be relatively safe when administered to oral, eye and skin of the animal models for short periods of time.


Quantification of cAMP and cGMP analogs in intact cells: pitfalls in enzyme immunoassays for cyclic nucleotides.
Immunoassays are routinely used as research tools to measure intracellular cAMP and cGMP concentrations. Ideally, this application requires antibodies with high sensitivity and specificity. The present work evaluates the cross-reactivity of commercially available cyclic nucleotide analogs with two non-radioactive and one radioactive cAMP and cGMP immunoassay. Most of the tested cyclic nucleotide analogs showed low degree competition with the antibodies; however, with Rp-cAMPS, 8-Br-cGMP and 8-pCPT-cGMP, a strong cross-reactivity with the corresponding cAMP and cGMP, respectively, immunoassays was observed. The determined EIA-binding constants enabled the measurement of the intracellular cyclic nucleotide concentrations and revealed a time- and lipophilicity-dependent cell membrane permeability of the compounds in the range of 10-30% of the extracellular applied concentration, thus allowing a more accurate prediction of the intracellular analog levels in a given experiment.


Treatment and outcome of patients suffering from perineal/perianal rhabdomyosarcoma: results from the CWS trials--retrospective clinical study.
To analyze the clinical course, treatment, complications, outcome, and quality of life (QOL) in patients with perineal/perianal rhabdomyosarcoma (PRMS) treated within the CWS-86, -91, -96, and -2002P trials. Although multiple international study trials exist for the treatment of rhabdomyosarcoma, only very limited information is given on treatment, outcome, and QOL in PRMS. A total of 35 patients suffering from PRMS were treated with neoadjuvant chemotherapy. Local therapy with radiation and/or surgery was performed, followed by adjuvant chemotherapy. Functional long-term follow-up was evaluated by a gastrointestinal/QOL survey. Thirty-two patients were evaluated (exclusion n = 3). Eight patients had embryonal histology, and 24 patients had alveolar histology. The median age was 108 months (median follow-up: 5.8 years). The 5-year overall survival was 47% (95% confidence interval: 29-64). Sixteen IRS (Intergroup Rhabdomyosarcoma Study) III and IV patients had locoregional lymph node involvement at diagnosis. Seven patients were treated with chemotherapy/surgery alone [5-year event-free survival (EFS): 85.7%]. Eleven patients received only radiochemotherapy (5-year EFS: 27.3%). Combined radiochemotherapy/surgery was used in 12 patients (5-year EFS: 63.6%). Two patients were treated only with chemotherapy and they died. Patients with embryonal histology had a significantly better 5-year EFS (87.5%) than patients with alveolar histology (39.1%; P = 0.013). Some patients reported symptoms of fecal incontinence. The median Wexner fecal incontinence score was 9 (possible range: 0-20), and the median QOL score was 90.5 (applicable range: 0-144). The outcome of these patients remains unsatisfactory. Prognostic factors for a favorable outcome are tumor size of smaller than 5 cm, negative locoregional lymph nodes, age less than 10 years, low IRS group, and embryonal histology. Fecal incontinence seems to be a problem.


The effect of night extension orthoses following surgical release of Dupuytren contracture: a single-center, randomized, controlled trial.
To clarify the efficacy and detrimental effects of orthoses used to maintain finger extension following surgical release of Dupuytren contracture. We conducted a single-center, randomized, controlled trial to investigate the effect of night extension orthoses on finger range of motion and hand function for 3 months following surgical release of Dupuytren contracture. We also wanted to determine how well finger extension was maintained in the total sample. We randomized 56 patients to receive a night extension orthosis plus hand therapy (n = 26) or hand therapy alone (n = 30). The primary outcome was total active extension of the operated fingers (°). Secondary outcomes were total active flexion of the operated fingers (°), active distal palmar crease (cm), grip strength (kg), and self-reported hand function using the Disabilities of the Arm, Shoulder, and Hand questionnaire (0-100 scale). There were no statistically significant differences between the no-orthosis and orthosis groups for total active extension or for any of the secondary outcomes. Between the first postoperative measure and 3 months after surgery, 62% of little fingers had maintained or improved total active extension. The use of a night extension orthosis in combination with standard hand therapy has no greater effect on maintaining finger extension than hand therapy alone in the 3 months following surgical release of Dupuytren contracture. Our results indicate that the practice of providing every patient with a night extension orthosis following surgical release of Dupuytren contracture may not be justified except for cases in which extension loss occurs after surgery. Our results also challenge clinicians to research ways of maintaining finger extension in a greater number of patients.


The acute patient physiologic and laboratory evaluation (APPLE) score: a severity of illness stratification system for hospitalized dogs.
Objective risk stratification models are used routinely in human critical care medicine. Applications include quantitative and objective delineation of illness severity for patients enrolled in clinical research, performance benchmarking, and protocol development for triage and therapeutic management. To develop an accurate, validated, and user-friendly model to stratify illness severity by mortality risk in hospitalized dogs. Eight hundred and ten consecutive intensive care unit (ICU) admissions of dogs at a veterinary teaching hospital. Prospective census cohort study. Data on 55 management, physiological, and biochemical variables were collected within 24 hours of admission. Data were randomly divided, with 598 patient records used for logistic regression model construction and 212 for model validation. Patient mortality was 18.4%. Ten-variable and 5-variable models were developed to provide both a high-performance model and model maximizing accessibility, while maintaining good performance. The 10-variable model contained creatinine, WBC count, albumin, SpO(2) , total bilirubin, mentation score, respiratory rate, age, lactate, and presence of free fluid in a body cavity. Area under the receiver operator characteristic (AUROC) on the construction data set was 0.93, and on the validation data set was 0.91. The 5-variable model contained glucose, albumin, mentation score, platelet count, and lactate. AUROC on the construction data set was 0.87, and on the validation data set was 0.85. Two models are presented that enable allocation of an accurate and user-friendly illness severity index for dogs admitted to an ICU. These models operate independent of primary diagnosis, and have been independently validated.


Polybrominated dibenzo-p-dioxins, dibenzofurans, and biphenyls: inclusion in the toxicity equivalency factor concept for dioxin-like compounds.
In 2011, a joint World Health Organization (WHO) and United Nations Environment Programme (UNEP) expert consultation took place, during which the possible inclusion of brominated analogues of the dioxin-like compounds in the WHO Toxicity Equivalency Factor (TEF) scheme was evaluated. The expert panel concluded that polybrominated dibenzo-p-dioxins (PBDDs), dibenzofurans (PBDFs), and some dioxin-like biphenyls (dl-PBBs) may contribute significantly in daily human background exposure to the total dioxin toxic equivalencies (TEQs). These compounds are also commonly found in the aquatic environment. Available data for fish toxicity were evaluated for possible inclusion in the WHO-UNEP TEF scheme (van den Berg et al., 1998). Because of the limited database, it was decided not to derive specific WHO-UNEP TEFs for fish, but for ecotoxicological risk assessment, the use of specific relative effect potencies (REPs) from fish embryo assays is recommended. Based on the limited mammalian REP database for these brominated compounds, it was concluded that sufficient differentiation from the present TEF values of the chlorinated analogues (van den Berg et al., 2006) was not possible. However, the REPs for PBDDs, PBDFs, and non-ortho dl-PBBs in mammals closely follow those of the chlorinated analogues, at least within one order of magnitude. Therefore, the use of similar interim TEF values for brominated and chlorinated congeners for human risk assessment is recommended, pending more detailed information in the future.


Monte Carlo study of the formation and conformational properties of dimers of Aβ42 variants.
Small soluble oligomers, and dimers in particular, of the amyloid β-peptide (Aβ) are believed to play an important pathological role in Alzheimer's disease. Here, we investigate the spontaneous dimerization of Aβ42, with 42 residues, by implicit solvent all-atom Monte Carlo simulations, for the wild-type peptide and the mutants F20E, E22G and E22G/I31E. The observed dimers of these variants share many overall conformational characteristics but differ in several aspects at a detailed level. In all four cases, the most common type of secondary structure is intramolecular antiparallel β-sheets. Parallel, in-register β-sheet structure, as in models for Aβ fibrils, is rare. The primary force driving the formation of dimers is hydrophobic attraction. The conformational differences that we do see involve turns centered in the 20-30 region. The probability of finding turns centered in the 25-30 region, where there is a loop in Aβ fibrils, is found to increase upon dimerization and to correlate with experimentally measured rates of fibril formation for the different Aβ42 variants. Our findings hint at reorganization of this part of the molecule as a potentially critical step in Aβ aggregation.


Improving the In Vivo Profile of Minigastrin Radiotracers: A Comparative Study Involving the Neutral Endopeptidase Inhibitor Phosphoramidon.
Minigastrin radiotracers, such as [(111)In-DOTA]MG0 ([(111)In-DOTA-DGlu(1)]minigastrin), have been considered for diagnostic imaging and radionuclide therapy of CCK2R-positive human tumors, such as medullary thyroid carcinoma. However, the high kidney retention assigned to the pentaGlu(2-6) repeat in the peptide sequence has compromised their clinical applicability. On the other hand, truncated des(Glu)(2-6)-analogs, such as [(111)In-DOTA]MG11 ([(111)In-DOTA-DGlu(10),desGlu(2-6)]minigastrin), despite their low renal uptake, show poor bioavailability and tumor targeting. [(111)In]CP04 ([(111)In-DOTA-DGlu(1-6)]minigastrin) acquired by Glu(2-6)/DGlu(2-6) substitution showed promising tumor-to-kidney ratios in rodents. In the present study, we compare the biological profiles of [(111)In]CP04, [(111)In-DOTA]MG11, and [(111)In-DOTA]MG0 during in situ neutral endopeptidase (NEP) inhibition, recently shown to improve the bioavailability of several peptide radiotracers. After coinjection of the NEP inhibitor, phosphoramidon (PA), the stability of [(111)In]CP04 and [(111)In-DOTA]MG0 in peripheral mouse blood increased, with an exceptional >14-fold improvement monitored for [(111)In-DOTA]MG11. In line with these findings, PA treatment increased the uptake of [(111)In]CP04 (8.5 ± 0.4%ID/g to 16.0 ± 2.3%ID/g) and [(111)In-DOTA]MG0 (11.9 ± 2.2%ID/g to 17.2 ± 0.9%ID/g) in A431-CCK2R(+) tumors at 4 hours postinjection, whereas the respective increase for [(111)In-DOTA]MG11 was >6-fold (2.5 ± 0.9%ID/g to 15.1 ± 1.7%ID/g). Interestingly, kidney uptake remained lowest for [(111)In-DOTA]MG11, but unfavorably increased by PA treatment for [(111)In-DOTA]MG0. Thus, overall, the most favorable in vivo profile was displayed by [(111)In-DOTA]MG11 during NEP inhibition, highlighting the need to validate this promising concept in the clinic.