Estimation of expected utility gained from the helmet law in Taiwan by quality-adjusted survival time.
The objective of this study was to empirically estimate the expected utility gained from the implementation of the 1997 helmet law in Taiwan by using quality-adjusted survival time (QAST). We randomly selected 400 out of 8221 registered cases of head injury and successfully interviewed 99 cases with the index of health-related quality of life (IHRQ). The function of IHRQ was then multiplied with the corresponding survival function to obtain the QAST for head injury. The total utility gained from the helmet law in Taipei within 1 year was estimated by multiplying the expected loss of utility per patient with the number of prevented cases. The results showed that after 80 months of follow-up, the QAST of the injured population was 66.3 quality-adjusted life-months (QALMs), while that of the reference population was 78.7 QALMs. We extrapolated the QAST for total life expectancy by simulating the survival of head injury cases using the life table data from the general population. The life-long utility loss of a head injury case was found to be 4.8 quality-adjusted life-years (QALY). The number of prevented cases during the first year of enforcement of the helmet law was estimated to be 1300 cases in Taipei, which amounted to 6240 QALYs gained. For lack of data, we were unable to calculate the possible gain from helmet on reduction of severity among nonfatal cases with head injury, and the estimation was only a lower bound. We concluded that the QAST approach is a feasible approach applicable to health policy decision-making, especially in cost-utility analysis.


Tomato Fruit Development and Metabolism.
Tomato (Solanum lycopersicum L.) belongs to the Solanaceae family and is the second most important fruit or vegetable crop next to potato (Solanum tuberosum L.). It is cultivated for fresh fruit and processed products. Tomatoes contain many health-promoting compounds including vitamins, carotenoids, and phenolic compounds. In addition to its economic and nutritional importance, tomatoes have become the model for the study of fleshy fruit development. Tomato is a climacteric fruit and dramatic metabolic changes occur during its fruit development. In this review, we provide an overview of our current understanding of tomato fruit metabolism. We begin by detailing the genetic and hormonal control of fruit development and ripening, after which we document the primary metabolism of tomato fruits, with a special focus on sugar, organic acid, and amino acid metabolism. Links between primary and secondary metabolic pathways are further highlighted by the importance of pigments, flavonoids, and volatiles for tomato fruit quality. Finally, as tomato plants are sensitive to several abiotic stresses, we briefly summarize the effects of adverse environmental conditions on tomato fruit metabolism and quality.


Sexual selection and the evolution of brain size in primates.
Reproductive competition among males has long been considered a powerful force in the evolution of primates. The evolution of brain size and complexity in the Order Primates has been widely regarded as the hallmark of primate evolutionary history. Despite their importance to our understanding of primate evolution, the relationship between sexual selection and the evolutionary development of brain size is not well studied. The present research examines the evolutionary relationship between brain size and two components of primate sexual selection, sperm competition and male competition for mates. Results indicate that there is not a significant relationship between relative brain size and sperm competition as measured by relative testis size in primates, suggesting sperm competition has not played an important role in the evolution of brain size in the primate order. There is, however, a significant negative evolutionary relationship between relative brain size and the level of male competition for mates. The present study shows that the largest relative brain sizes among primate species are associated with monogamous mating systems, suggesting primate monogamy may require greater social acuity and abilities of deception.


Probing the lateral organization of membranes: fluorescence repercussions of pyrene probe distribution.
Phospholipids pyrene labeled are widely used to investigate dynamics and organizations of membranes. We studied pyrene probe lateral distribution by analyzing the variations of the molar absorption coefficient (epsilon) versus probe concentrations, in small unilamellar vesicles (SUV) made of phospholipids and/or glycolipids, with pyrene labeled phosphatidylcholine (PyPC) or phosphatidylglycerol (PyPG). The results were interpreted according to an infinite associative model. They indicated that an effective self-association process corresponding to K ranging from 30 to 100 M(-1) occurred with those probes incorporated in dimannosyl diacylglycerol (DMDG). In contrast, after SUV labeling of egg yolk phosphatidylcholine (EggPC) or phosphatidylglycerol (EggPG), K values < 1 M(-1) were determined. The corresponding percentages of various stacked forms of pyrene probes were calculated. They indicated that, for a 3% PyPG labeling, the monomer represented 21% of n-mers in DMDG and 94% in EggPC. The analysis of fluorescence experiments carried out on the same samples indicated that: (i) the fluorescence process of pyrene probes was generated by the monomers: and (ii) the excimer forming resulted from a diffusional encounter between one excited and one non-excited monomer. A correction of fluorescence data allowing a more correct interpretation of fluorescence measurements was proposed.


High-numerical-aperture microscopy with a rotating point spread function.
Rotating point spread function (PSF) microscopy via spiral phase engineering can localize point sources over large focal depths in a snapshot mode. The present work gives an approximate vector-field analysis of an improved rotating PSF design that encodes both the 3D location and polarization state of a monochromatic point dipole emitter for high-numerical-aperture microscopy. By examining the angle of rotation and the spatial form of the PSF, one can jointly localize point sources and determine the polarization state of light emitted by them over a 3D field in a single snapshot. Results of numerical simulations of noisy data frames under Poisson shot noise conditions and the errors in the recovery of 3D location and dipole orientation for a single point source are discussed.


Differential activation of virulence gene expression by PrfA, the Listeria monocytogenes virulence regulator.
PrfA is a pleiotropic activator of virulence gene expression in the pathogenic bacterium Listeria monocytogenes. Several lines of evidence have suggested that a hierarchy of virulence gene activation by PrfA exists. This hypothesis was investigated by assessing the ability of PrfA to activate the expression of virulence gene fusions to lacZ in Bacillus subtilis. Expression of PrfA in this heterologous host was sufficient for activation of transcription at the hly, plcA, mpl, and actA promoters. Activation was most efficient at the divergently transcribed hly and plcA promoters. The putative PrfA binding site shared by these promoters is perfectly symmetrical and appears to represent the optimum sequence for target gene activation by PrfA. The activation of actA and mpl expression was considerably weaker and occurred more slowly than that observed at the hly and plcA promoters, suggesting that greater quantities of PrfA are required for productive interaction at these promoters. Interestingly, expression of an inlA-lacZ transcriptional fusion was very poorly activated by PrfA in B. subtilis, suggesting that other Listeria factors, in addition to PrfA, are required for PrfA-mediated activation at this promoter. Further support for the involvement of such factors was obtained by constructing and analyzing a prfA deletion mutant of L. monocytogenes. We observed that, in contrast to that of the other genes of the PrfA regulon, expression of inlA is only partially dependent on PrfA.


Reassessment of the 2010-2011 Haiti cholera outbreak and rainfall-driven multiseason projections.
Mathematical models can provide key insights into the course of an ongoing epidemic, potentially aiding real-time emergency management in allocating health care resources and by anticipating the impact of alternative interventions. We study the ex post reliability of predictions of the 2010-2011 Haiti cholera outbreak from four independent modeling studies that appeared almost simultaneously during the unfolding epidemic. We consider the impact of different approaches to the modeling of spatial spread of Vibrio cholerae and mechanisms of cholera transmission, accounting for the dynamics of susceptible and infected individuals within different local human communities. To explain resurgences of the epidemic, we go on to include waning immunity and a mechanism explicitly accounting for rainfall as a driver of enhanced disease transmission. The formal comparative analysis is carried out via the Akaike information criterion (AIC) to measure the added information provided by each process modeled, discounting for the added parameters. A generalized model for Haitian epidemic cholera and the related uncertainty is thus proposed and applied to the year-long dataset of reported cases now available. The model allows us to draw predictions on longer-term epidemic cholera in Haiti from multiseason Monte Carlo runs, carried out up to January 2014 by using suitable rainfall fields forecasts. Lessons learned and open issues are discussed and placed in perspective. We conclude that, despite differences in methods that can be tested through model-guided field validation, mathematical modeling of large-scale outbreaks emerges as an essential component of future cholera epidemic control.


Effect of different angles of knee flexion on leg extensor power in healthy individuals.
To investigate the effect of fixed flexion knee deformity on leg extensor power as measured by the Nottingham leg extensor power rig. Cross-sectional observational study. Orthopaedic hospital. A convenience sample of 135 adult participants. leg extensor power normalised for body weight, UCLA activity scale. Power values at 0° FF were found to be significantly less than power values at 15° FF [difference 0.21W/kg SD .36], and power values at 15° FF were significantly less than those at 30° FF [difference 0.31W/kg SD .43; P<0.001) in both right and left legs. Age and activity levels were moderately negatively correlated, with UCLA score decreasing with increasing age (-0.343, P<0.0005). No significant correlation was found between activity levels measured on the UCLA and power on the LEP. Given the large range of pre-operative maximal extension, the validity of testing each patient at their own maximal range of pre-intervention extension and then at the same angle post-intervention is questionable. In studies assessing change in power following an intervention, the end point angle should be standardised between individuals. In future studies investigating leg extensor power on the LEP rig pre and post intervention, it could be more appropriate to standardise the angle of FF to 30°, with individuals who are unable to achieve this position excluded from the study.


Molecular and immunological characterisation of Theileria parva stocks which are components of the 'Muguga cocktail' used for vaccination against East Coast fever in cattle.
The 'Muguga cocktail' which is composed of three Theileria parva stocks Muguga, Kiambu 5 and Serengeti-transformed has been used extensively for live vaccination against East Coast fever in cattle in eastern, central and southern Africa. Herein we describe the molecular characterisation of the T. parva vaccine stocks using three techniques, an indirect fluorescent antibody test with a panel of anti-schizont monoclonal antibodies (MAb), Southern blotting with four T. parva repetitive DNA probes and polymerase chain reaction (PCR)-based assays detecting polymorphism within four single copy loci encoding antigen genes. The Muguga and Serengeti-transformed stocks exhibited no obvious differences in their reactivity with the panel of MAbs, whereas Kiambu 5 differed with several MAbs. Kiambu 5 DNA was very distinct from the Muguga and Serengeti-transformed isolates in the hybridisation pattern with all four nucleic acid probes, whereas Muguga and Serengeti-transformed isolates exhibited minor differences and could not be discriminated with one of the probes. PCR amplification in combination with restriction fragment length polymorphism analysis indicated that Kiambu 5 was also markedly divergent from the Muguga and Serengeti-transformed stocks within two of the four antigen coding genes. The T. parva Serengeti-transformed stock did not contain a 130 base pair insert within the p67 sporozoite antigen gene, which has been observed previously in most T. parva parasites isolated from buffalo, and could not be discriminated from T. parva Muguga at any of the four single copy loci. Collectively the data indicate that two of the cocktail components T. parva Serengeti-transformed and Muguga are genetically closely related, while the third component Kiambu 5 is quite distinct. Based on the findings, there may be a need to include only one of the T. parva Muguga and Serengeti-transformed components in the immunising cocktail. The study demonstrates the value of molecular characterisation data for monitoring of live vaccines.


Analysis of the FHIT gene and FRA3B region in sporadic breast cancer, preneoplastic lesions, and familial breast cancer probands.
The FHIT gene, which spans the FRA3B fragile site at chromosome 3p14.2, is a candidate tumor suppressor gene in breast and other cancers. We investigated FHIT and FRA3B for loss of heterozygosity (LOH); homozygous deletions; abnormal transcripts; and acquired/germ-line point mutations in breast cancer cell lines (n = 32), breast epithelial and stromal cell cultures (n = 18), microdissected invasive (n = 16) and ductal in situ carcinomas (n = 6), and their accompanying normal and abnormal epithelial foci (n = 14). LOH at 3p14.2, especially at FHIT intragenic marker D3S1300, was found in 6 of 16 microdissected invasive tumors and 3 of 6 ductal in situ carcinomas. In accompanying preneoplastic foci, LOH occurred in two of eight intraductal hyperplasias but not in histologically normal ductal epithelium (n = 6). Three of 32 (9%) breast cancer cell lines demonstrated homozygous deletions of FHIT exon 4 (two cases) and exon 5 (one case), which correlated with exon 4-deleted transcripts and loss of the cDNA transcript containing the coding exons 5-9, respectively. Normal mammary cultures and 31 of 32 tumor cell lines (97%) expressed wild-type coding transcripts as well as a minor exon 8-deleted message. Single-strand conformation polymorphism analysis of the coding exons in the 32 tumor and 18 normal breast cell lines and their sequencing revealed four silent polymorphisms and a germ-line histidine triad point mutation (651 G-->T) in a tumor arising in a 70-year-old woman. This mutation was also present in one of her two thus far unaffected daughters. Analysis of additional DNAs from 280 probands of high-risk breast cancer families for other FHIT exon 8 mutations detected an intronic point mutation 13 bases upstream of exon 8. Thus, we have demonstrated relatively early abnormalities of the FHIT/FRA3B region in breast cancer and discovered two rare FHIT germ-line mutations. The expression of a transcript containing the coding exons in nearly all cell lines, including those with germ-line mutations, suggests the possibility that another gene in the FRA3B region may be involved in the pathogenesis of breast cancer.


Thermal conductivity of highly porous Si in the temperature range 4.2 to 20 K.
We report on experimental results of the thermal conductivity k of highly porous Si in the temperature range 4.2 to 20 K, obtained using the direct current (dc) method combined with thermal finite element simulations. The reported results are the first in the literature for this temperature range. It was found that porous Si thermal conductivity at these temperatures shows a plateau-like temperature dependence similar to that obtained in glasses, with a constant k value as low as 0.04 W/m.K. This behavior is attributed to the presence of a majority of non-propagating vibrational modes, resulting from the nanoscale fractal structure of the material. By examining the fractal geometry of porous Si and its fractal dimensionality, which was smaller than two for the specific porous Si material used, we propose that a band of fractons (the localized vibrational excitations of a fractal lattice) is responsible for the observed plateau. The above results complement previous results by the authors in the temperature range 20 to 350 K. In this temperature range, a monotonic increase of k with temperature is observed, fitted with simplified classical models. The extremely low thermal conductivity of porous Si, especially at cryogenic temperatures, makes this material an excellent substrate for Si-integrated microcooling devices (micro-coldplate). 61.43.-j; 63.22.-m; 65.8.-g.


Issues and needs in end-of-life decision making: an international modified Delphi study.
end-of-life decision making is an important aspect of end-of-life care that can have a significant impact on the process of dying and patients' comfort in the last days of life. the aim of our study was to identify issues and considerations in end-of-life decision making, and needs for more evidence among palliative care experts, across countries and professions. 90 palliative care experts from nine countries participated in a modified Delphi study. Participants were asked to identify important issues and considerations in end-of-life decision making and to rate the need for more evidence. experts mentioned 219 issues in end-of-life decision making related to the medical domain, 122 issues related to the patient wishes and 92 related to relatives' wishes, regardless of profession or country (p > 0.05). In accordance, more than 90% of the experts rated the comfort and wishes of the patient and the potential futility of treatment as important considerations in end-of-life decision making, although some variation was present. When asked about issues that are in need of more evidence, 87% mentioned appropriate indications for using sedatives and effects of artificial hydration at the end of life. A total of 83% mentioned adequate communication approaches. palliative care experts from different professions in different countries encounter similar issues in end-of-life decision making. Adequate communication about these issues is universally experienced as a challenge, which might benefit from increased knowledge. This shared experience enables and emphasizes the need for more international research.


Sub-threshold depressive symptoms and brain structure: A magnetic resonance imaging study within the Whitehall II cohort.
Late-life sub-threshold depressive symptoms (i.e. depressive symptoms that do not meet the criteria for a diagnosis of major depressive disorder) are associated with impaired physical health and function, and increased risk of major depressive disorder. Magnetic resonance imaging (MRI) studies examining late-life major depressive disorder find structural brain changes in grey and white matter. However, the extent to which late-life sub-threshold depression is associated with similar hallmarks is not well established. Participants with no history of major depressive disorder were selected from the Whitehall Imaging Sub-Study (n=358, mean age 69±5 years, 17% female). Depressive symptoms were measured using the Centre for Epidemiological Studies Depression Scale (CES-D) at three previous Whitehall II Study phases (2003-04, 2007-09 and 2012-13) and at the time of the MRI scan (2012-14). The relationships between current and cumulative depressive symptoms and MRI brain measures were explored using Voxel-Based Morphometry (VBM) for grey matter and Tract Based Spatial Statistics (TBSS) for white matter. Current sub-threshold depressive symptoms were associated with significant reductions in fractional anisotropy and increases in axial and radial diffusivity. There were no significant relationships between current depressive symptoms and grey matter measures, or cumulative depressive symptoms and MRI measures. The prevalence (10%) of sub-threshold depressive symptoms means that analyses may be underpowered to detect subtle differences in brain structure. Current sub-threshold depressive symptoms are associated with changes in white matter microstructure, indicating that even mild depressive symptoms are associated with similar MRI hallmarks to those in major depressive disorder.


The ble resistance gene as a new selectable marker for Trypanosoma brucei: fly transmission of stable procyclic transformants to produce antibiotic resistant bloodstream forms.
We describe here the stable transformation of Trypanosoma brucei using a new selectable marker for kinetoplastid protozoa, the Sh ble, or phleomycin, resistance gene. A plasmid containing this gene targeted to the tubulin gene locus by homologous sequences was introduced into procyclic trypanosomes by electroporation and cells selected for antibiotic resistance. Southern analysis of stable transformants showed that the plasmid had been integrated into the tubulin locus by homologous recombination. Analysis of bloodstream stage transformants, produced by transmission through the vector Glossina, showed that the resistance gene was conserved and expressed in these forms in the absence of selective drug pressure. In both procyclic and bloodstream forms, transcription of the ble gene appears to originate from the upstream tubulin promoter, despite the presence of a VSG promoter in the integrated construct. The generation of stable bloodstream transformants for the first time will facilitate the study of gene function and expression during the trypanosome life cycle, and aid in the investigation of genetic exchange in these organisms.


Chromosome arm length and nuclear constraints determine the dynamic relationship of yeast subtelomeres.
Physical interactions between distinct chromosomal genomic loci are important for genomic functions including recombination and gene expression, but the mechanisms by which these interactions occur remain obscure. Using telomeric association as a model system, we analyzed here the in vivo organization of chromosome ends of haploid yeast cells during interphase. We separately labeled most of the 32 subtelomeres and analyzed their positions both in nuclear space and relative to three representative reference subtelomeres by high-throughput 3D microscopy and image processing. We show that subtelomeres are positioned nonrandomly at the nuclear periphery, depending on the genomic size of their chromosome arm, centromere attachment to the microtubule organizing center (spindle pole body, SPB), and the volume of the nucleolus. The distance of subtelomeres to the SPB increases consistently with chromosome arm length up to approximately 300 kb; for larger arms the influence of chromosome arm length is weaker, but the effect of the nucleolar volume is stronger. Distances between pairs of subtelomeres also exhibit arm-length dependence and suggest, together with dynamic tracking experiments, that potential associations between subtelomeres are unexpectedly infrequent and transient. Our results suggest that interactions between subtelomeres are nonspecific and instead governed by physical constraints, including chromosome structure, attachment to the SPB, and nuclear crowding.


Universal resilience patterns in complex networks.
Resilience, a system's ability to adjust its activity to retain its basic functionality when errors, failures and environmental changes occur, is a defining property of many complex systems. Despite widespread consequences for human health, the economy and the environment, events leading to loss of resilience--from cascading failures in technological systems to mass extinctions in ecological networks--are rarely predictable and are often irreversible. These limitations are rooted in a theoretical gap: the current analytical framework of resilience is designed to treat low-dimensional models with a few interacting components, and is unsuitable for multi-dimensional systems consisting of a large number of components that interact through a complex network. Here we bridge this theoretical gap by developing a set of analytical tools with which to identify the natural control and state parameters of a multi-dimensional complex system, helping us derive effective one-dimensional dynamics that accurately predict the system's resilience. The proposed analytical framework allows us systematically to separate the roles of the system's dynamics and topology, collapsing the behaviour of different networks onto a single universal resilience function. The analytical results unveil the network characteristics that can enhance or diminish resilience, offering ways to prevent the collapse of ecological, biological or economic systems, and guiding the design of technological systems resilient to both internal failures and environmental changes.


Mixed impact of firearms restrictions on fatal firearm injuries in males: a national observational study.
Public health organizations have recommended restricted access and safe storage practices as means to reduce firearm injuries and deaths. We aimed to assess the effect of four firearm restrictions on firearm deaths in Norway 1969-2009. All deaths due to firearm discharge were included (5,660 deaths, both sexes). The statistical analysis to assess impact of firearm legislations was restricted to males because of the sex disproportionality (94% were males). A total of 89% of firearm deaths (both sexes) were classified as suicide, 8% as homicide, and 3% as unintentional (accident). During the past four decades, male accidental firearm death rates were reduced significantly by 90%. Male firearms suicide rates increased from 1969 to 1991 by 166%, and decreased by 62% from 1991 to 2009. Despite the great reduction in male accidental firearm deaths, we were unable to demonstrate effects of the laws. In contrast, we found that a 1990 regulation, requiring a police permit before acquiring a shotgun, had a beneficial impact on suicide in the total sample and in those aged 15-34 years. Male firearm homicides decreased post-2003 regulation regarding storing home guard weapons in private homes. Our findings suggest that two laws could have contributed to reduce male firearm mortality. It is, however, a challenge to measure the role of four firearm restrictions. The null findings are inconclusive, as they may reflect no true impact or study limitations.


Topography of cortical thinning areas associated with hippocampal atrophy (HA) in patients with Alzheimer's disease (AD).
Prior studies have shown that patients with AD have decreased functional or structural connectivity between the hippocampus and other brain areas. To the best of our knowledge, however, there have been no studies investigating the topography of cortical thinning areas and correlations with HA using surface based morphometry of three-dimensional (3D) T1-weighted magnetic resonance (MR) images. Cortical thickness was measured using SBM, and hippocampal volume was measured using an automated method, in 219 patients with AD and 54 subjects with no cognitive impairment (NCI). A partial correlation model was used in analysis of cortical thinning related to HA. Cortical thinning areas related to HA were found mostly within areas associated with polysynaptic or direct pathways of the hippocampus, a finding consistent with the disconnection hypothesis. Therefore, the cortical atrophy related to HA in patients with AD may represent disrupted cortical brain networks in connection with HA. However, since the topography of HA-related cortical thinning in groups with Clinical Dementia Ratings (CDR) of 0.5 and 1 corresponded to the stages I-II and III-IV of Braak and Braak staging, respectively, we could not exclude the possibility of the "concomitant hypothesis," i.e. that these areas are affected concomitantly with the hippocampus.


A spike sorting toolbox for up to thousands of electrodes validated with ground truth recordings in vitro and in vivo.
In recent years, multielectrode arrays and large silicon probes have been developed to record simultaneously between hundreds and thousands of electrodes packed with a high density. However, they require novel methods to extract the spiking activity of large ensembles of neurons. Here, we developed a new toolbox to sort spikes from these large-scale extracellular data. To validate our method, we performed simultaneous extracellular and loose patch recordings in rodents to obtain 'ground truth' data, where the solution to this sorting problem is known for one cell. The performance of our algorithm was always close to the best expected performance, over a broad range of signal-to-noise ratios, in vitro and in vivo. The algorithm is entirely parallelized and has been successfully tested on recordings with up to 4225 electrodes. Our toolbox thus offers a generic solution to sort accurately spikes for up to thousands of electrodes.


Associations between Plasma Choline Metabolites and Genetic Polymorphisms in One-Carbon Metabolism in Postmenopausal Women: The Women's Health Initiative Observational Study.
Choline plays an integral role in one-carbon metabolism in the body, but it is unclear whether genetic polymorphisms are associated with variations in plasma choline and its metabolites. This study aimed to evaluate the association of genetic variants in choline and one-carbon metabolism with plasma choline and its metabolites. We analyzed data from 1423 postmenopausal women in a case-control study nested within the Women's Health Initiative Observational Study. Plasma concentrations of choline, betaine, dimethylglycine (DMG), and trimethylamine N-oxide were determined in 12-h fasting blood samples collected at baseline (1993-1998). Candidate and tagging single-nucleotide polymorphisms (SNPs) were genotyped in betaine-homocysteine S-methyltransferase (BHMT), BHMT2, 5,10-methylenetetrahydrofolate reductase (MTHFR), methylenetetrahydrofolate dehydrogenase (NADP+ dependent 1) (MTHFD1), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and 5-methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR). Linear regression was used to derive percentage difference in plasma concentrations per variant allele, adjusting for confounders, including B-vitamin biomarkers. Potential effect modification by plasma vitamin B-12, vitamin B-6, and folate concentrations and folic-acid fortification periods was examined. The candidate SNP BHMT R239Q (rs3733890) was associated with lower concentrations of plasma betaine and DMG concentrations (-4.00% and -6.75% per variant allele, respectively; both nominal P < 0.05). Another candidate SNP, BHMT2 rs626105 A>G, was associated with higher plasma DMG concentration (13.0%; P < 0.0001). Several tagSNPs in these 2 genes were associated with plasma concentrations after correction for multiple comparisons. Vitamin B-12 status was a significant effect modifier of the association between the genetic variant BHMT2 rs626105 A>G and plasma DMG concentration. Genetic variations in metabolic enzymes were associated with plasma concentrations of choline and its metabolites. Our findings contribute to the knowledge on the variation in blood nutrient concentrations in postmenopausal women.


Quorum Sensing by Monocyte-Derived Populations.
Quorum sensing is a type of cellular communication that was first described in bacteria, consisting of gene expression regulation in response to changes in cell-population density. Bacteria synthesize and secrete diffusive molecules called autoinducers, which concentration varies accordingly with cell density and can be detected by the producing cells themselves. Once autoinducer concentration reaches a critical threshold, all bacteria within the autoinducer-rich environment react by modifying their genetic expression and adopt a coordinated behavior (e.g., biofilm formation, virulence factor expression, or swarming motility). Recent advances highlight the possibility that such type of communication is not restricted to bacteria, but can exist among other cell types, including immune cells and more specifically monocyte-derived cells (1). For such cells, quorum sensing mechanisms may not only regulate their population size and synchronize their behavior at homeostasis but also alter their activity and function in unexpected ways during immune reactions. Although the nature of immune autoinducers and cellular mechanisms remains to be fully characterized, quorum sensing mechanisms in the immune system challenge our traditional conception of immune cell interactions and likely represent an important mode of communication at homeostasis or during an immune response. In this mini-review, we briefly present the prototypic features of quorum sensing in bacteria and discuss the existing evidence for quorum sensing within the immune system. Mainly, we review quorum sensing mechanisms among monocyte-derived cells, such as the regulation of inflammation by the density of monocyte-derived cells that produce nitric oxide and discuss the relevance of such models in the context of immune-related pathologies.


Developmental toxicity of benzyl benzoate in rats after maternal exposure throughout pregnancy.
The maternal and fetal toxicity of benzyl benzoate, commonly used as antiparasitic insecticide, was evaluated in pregnant rats after a daily oral dose of 25 and 100 mg/kg. Biochemical, histopathological, and morphological examinations were performed. Dams were observed for maternal body weights and food and water consumption and subjected to caesarean section on (GD) 20. Maternal and fetal liver, kidney, heart, brain, and placenta were examined histopathologically under light microscope. Maternal and fetal liver and placenta were stained immunohistochemically for vascular endothelial growth factor (VEGF). Morphometric analysis of fetal body lengths, placental measurements, and fetal skeletal stainings was performed. Statistically significant alterations in biochemical parameters and placental and skeletal measurements were determined in treatment groups. In addition to histopathological changes, considerable differences were observed in the immunolocalization of VEGF in treatment groups. These results demonstrated that benzyl benzoate and its metabolites can transport to the placenta and eventually enter the fetuses.


Origins and evolution of the Western diet: health implications for the 21st century.
There is growing awareness that the profound changes in the environment (eg, in diet and other lifestyle conditions) that began with the introduction of agriculture and animal husbandry approximately 10000 y ago occurred too recently on an evolutionary time scale for the human genome to adjust. In conjunction with this discordance between our ancient, genetically determined biology and the nutritional, cultural, and activity patterns of contemporary Western populations, many of the so-called diseases of civilization have emerged. In particular, food staples and food-processing procedures introduced during the Neolithic and Industrial Periods have fundamentally altered 7 crucial nutritional characteristics of ancestral hominin diets: 1) glycemic load, 2) fatty acid composition, 3) macronutrient composition, 4) micronutrient density, 5) acid-base balance, 6) sodium-potassium ratio, and 7) fiber content. The evolutionary collision of our ancient genome with the nutritional qualities of recently introduced foods may underlie many of the chronic diseases of Western civilization.


Floristic variation in plant communities on metalliferous mining residues in the northern and southern Pennines, England.
Vegetation and soil surveys were conducted on metalliferous mine wastes in the northern and southern Pennines of England. Analyses of vegetation composition in relation to soil chemical variation were performed. Ordination analysis facilitated the detection of groups of co-occurring species which are characteristic of types of metal-contaminated soil. The results of regression analysis implied the importance of soil pH and concentration of available lead or zinc, depending upon region, in determining species distributions. A strong interactive effect was evident between soil phosphorus and zinc content in influencing species distributions in the southern Pennines sample.Regional similarities in the vegetation of apparently similar metalliferous soils existed. The unusually high species richness of some soils was associated with relatively low concentrations of heavy metals. Many species of relatively floristically-rich wastes were also colonists of surrounding grasslands or woodland. This indicated the potential importance of propagule availability and capability for rapid establishment on bare or unstable ground. Mechanisms of physiological stress avoidance, rather than heavy-metai tolerance, may explain the occurrence of non-metallophytes on soils contaminated with lead and zinc.


15N isotopic labelling for in-cell protein studies by NMR spectroscopy and single-cell IR synchrotron radiation FTIR microscopy: a correlative study.
The ultimate goal of modern structural biology is to probe protein structures and dynamics in their physiological microenvironment. In-cell NMR spectroscopy is an ideal technique for achieving this goal, being able to investigate proteins at atomic-resolution in living cells. The reliability of the results provided by in-cell NMR relies on the selectivity of the labelling methodology coupled with the filtering capabilities offered by heteronuclear NMR experiments. However, solution NMR is not well-suited either for measuring to what extent the non-specific labelling occurs, or to evaluate how it is affected by cell-to-cell variability and, eventually, whether the labelling procedure affects the cellular macromolecular content in general. To answer these questions, we correlated in-cell 1D 1H and 2D 1H-15N NMR experiments on HEK293T cells overexpressing superoxide dismutase 1 (SOD1) with single-cell Synchrotron Radiation FTIR Microscopy (FTIRM) experiments on the same samples. We verified that SOD1 overexpression in 15N-enriched media does not induce modifications in the overall cellular profile, and that the cell-to-cell labelling variability is independent of SOD1 overexpression and is likely cell cycle-related. We concluded that the non-specific incorporation of 15N into cellular components other than the protein of interest is one of the main factors that hinder the possibility of in-cell conformational studies by FTIRM at the single-cell level. Improving labelling selectivity by employing protein insertion approaches, and increasing FTIRM sensitivity by plasmonic enhancement, would open new perspectives for in-cell ultra-sensitive single-protein conformational studies complementing NMR and vibrational analyses.


Aberrant expression of a disintegrin and metalloproteinase 17/tumor necrosis factor-alpha converting enzyme increases the malignant potential in human pancreatic ductal adenocarcinoma.
A disintegrin and metalloproteinase (ADAM) molecules are known for their unique potential to combine adhesion, proteolysis, and signaling. To understand the role of ADAM17/tumor necrosis factor-alpha (TNF-alpha) converting enzyme (TACE) in pancreatic ductal adenocarcinoma (PDAC), we investigated its expression, function, and in vitro regulation. ADAM17/TACE mRNA was expressed in 3 of 10 normal pancreatic tissues, 6 of 8 samples from patients with chronic pancreatitis, 10 of 10 PDAC tissues, and 9 of 9 pancreatic cancer cell lines, but it was absent in primary duct epithelial cells. Immunohistochemical staining revealed positive cancer cells in 8 of 10 PDACs but no staining of ducts in normal pancreas. ADAM17/TACE was found in 0 of 16 pancreatic intraepithelial neoplasia (PanIN)-1A lesions, 1 of 30 PanIN-1B lesions, 2 of 13 PanIN-2 lesions but, in 13 of 15 PanIN-3 lesions, associated with PDAC. Western blot, flow cytometry, and confocal microscopy analyses showed the aberrant expression of ADAM17/TACE protein in pancreatic cancer cell lines. The proteolytic activity of ADAM17/TACE, assessed by the release of TNF-alpha, was inhibited by TNF-alpha protease inhibitor. ADAM17/TACE gene silencing using small interfering RNA technique in vitro reduced invasion behavior dramatically, whereas proliferation was unaffected. Furthermore, ADAM17/TACE mRNA expression was down-regulated in pancreatic cancer cells arrested in G2-M phase as well as in a time-dependent manner after TNF-alpha and interleukin-6 incubation. In conclusion, our findings provide evidence of aberrant expression of the proteolytically active ADAM17/TACE in advanced precursor lesions (PanIN-3) and PDAC while identifying its critical involvement in the invasion process.


Retinoid-enhanced gap junctional communication is achieved by increased levels of connexin 43 mRNA and protein.
Natural and synthetic retinoids are potent inhibitors of experimental carcinogenesis in animals and cause reversion of premalignant lesions in humans. In the model C3H 10T1/2 cell system, retinoids enhance postconfluent growth control, reversibly inhibit carcinogen-induced transformation, and enhance gap junctional intercellular communication. These effects are highly correlated. 10T1/2 cells were found to express low levels of connexin 43, a gap junctional protein first found in the heart. After treatment of confluent 10T1/2 cells with the synthetic retinoid tetrahydrotetramethylnapthalenylpropenylbenzoic acid (TTNPB), levels of connexin 43 mRNA and protein increased within 6 h of treatment, while elevation of junctional communication was detected within 12-18 h. The maximally effective concentration of TTNPB (10(-8) M) caused an approximate 10-fold elevation of connexin 43 gene transcripts after 72 h. Indirect immunofluorescence microscopy using a polyclonal antibody to the synthetic C-terminal region of connexin 43 demonstrated that TTNPB induced many fluorescent plaques in regions of cell-cell contact. These results provide a molecular basis for the retinoid-enhanced junctional communication in 10T1/2 cells. It is proposed that one action of retinoids is to modulate the intercellular transfer of signal molecules. These could mediate many of the physiological actions of retinoids on growth control and carcinogenesis.


Serum albumin and kidney function decline in HIV-infected women.
Serum albumin concentrations are a strong predictor of mortality and cardiovascular disease in human immunodeficiency virus (HIV)-infected individuals. We studied the longitudinal associations between serum albumin levels and kidney function decline in a population of HIV-infected women. Retrospective cohort analysis. Study participants were recruited from the Women's Interagency HIV Study (WIHS), a large observational study designed to understand risk factors for the progression of HIV infection in women living in urban communities. 908 participants had baseline assessment of kidney function and 2 follow-up measurements over an average of 8 years. The primary predictor was serum albumin concentration. We examined annual change in kidney function. Secondary outcomes included rapid kidney function decline and incident reduced estimated glomerular filtration rate (eGFR). Kidney function decline was determined by cystatin C-based (eGFR(cys)) and creatinine-based eGFR (eGFR(cr)) at baseline and follow-up. Each model was adjusted for kidney disease and HIV-related risk factors using linear and relative risk regression. After multivariate adjustment, each 0.5-g/dL decrement in baseline serum albumin concentration was associated with a 0.56-mL/min faster annual decline in eGFR(cys) (P < 0.001), which was attenuated only slightly to 0.55 mL/min/1.73 m(2) after adjustment for albuminuria. Results were similar whether using eGFR(cys) or eGFR(cr). In adjusted analyses, each 0.5-g/dL lower baseline serum albumin level was associated with a 1.71-fold greater risk of rapid kidney function decline (P < 0.001) and a 1.72-fold greater risk of incident reduced eGFR (P < 0.001). The cohort is composed of only female participants from urban communities within the United States. Lower serum albumin levels were associated strongly with kidney function decline and incident reduced eGFRs in HIV-infected women independent of HIV disease status, body mass index, and albuminuria.


In vivo cell-autonomous transcriptional abnormalities revealed in mice expressing mutant huntingtin in striatal but not cortical neurons.
Huntington's disease (HD), caused by a CAG repeat expansion in the huntingtin (HTT) gene, is characterized by abnormal protein aggregates and motor and cognitive dysfunction. Htt protein is ubiquitously expressed, but the striatal medium spiny neuron (MSN) is most susceptible to dysfunction and death. Abnormal gene expression represents a core pathogenic feature of HD, but the relative roles of cell-autonomous and non-cell-autonomous effects on transcription remain unclear. To determine the extent of cell-autonomous dysregulation in the striatum in vivo, we examined genome-wide RNA expression in symptomatic D9-N171-98Q (a.k.a. DE5) transgenic mice in which the forebrain expression of the first 171 amino acids of human Htt with a 98Q repeat expansion is limited to MSNs. Microarray data generated from these mice were compared with those generated on the identical array platform from a pan-neuronal HD mouse model, R6/2, carrying two different CAG repeat lengths, and a relatively high degree of overlap of changes in gene expression was revealed. We further focused on known canonical pathways associated with excitotoxicity, oxidative stress, mitochondrial dysfunction, dopamine signaling and trophic support. While genes related to excitotoxicity, dopamine signaling and trophic support were altered in both DE5 and R6/2 mice, which may be either cell autonomous or non-cell autonomous, genes related to mitochondrial dysfunction, oxidative stress and the peroxisome proliferator-activated receptor are primarily affected in DE5 transgenic mice, indicating cell-autonomous mechanisms. Overall, HD-induced dysregulation of the striatal transcriptome can be largely attributed to intrinsic effects of mutant Htt, in the absence of expression in cortical neurons.


The use of radiography, urodynamic studies and cystoscopy in the evaluation of voiding dysfunction.
Children with dysfunctional voiding disorder often undergo radiological, cystoscopic or urodynamic evaluation to identify an anatomical or organic cause. We determined the role of these studies in the evaluation, management and ultimate outcome of a large patient population with voiding dysfunction at a single institution. We retrospectively evaluated the records of 1, 153 children with dysfunctional voiding disorder treated from 1990 to 1999. A thorough history and physical examination with specific emphasis on voiding patterns were done and urinalysis was performed in all cases. Ultrasound of the urinary system and excretory urography were done in 1,050 (91%) and 24 (2%) patients, respectively, while voiding cystourethrography was performed in 672 (58%), including 564 with a history of nonfebrile urinary tract infection. Cystoscopy and a formal urodynamic study were performed in 61 (5%) and 40 (3.5%) cases, respectively. Mean patient age at referral was 6 years (range 3 to 14). Of the children 74% were girls and 26% were boys. Physical examination of the abdomen, back, genitalia and neurological system was unremarkable in all cases. Ultrasound of the upper urinary system was normal in 1,018 patients (97%) and showed insignificant pyelectasis in 32 (3%). All 24 excretory urography studies were normal and voiding cystourethrography was normal in 470 of 672 cases (70%). Unilateral and bilateral low grade, and unilateral high grade reflux was present in 108, 19 and 3 patients, respectively. Urodynamic studies were performed in 40 children who did not respond to standard treatment. We noted detrusor instability in 16 patients, detrusor-sphincter dyssynergia in 6 and sensory abnormality in 3, while the study was completely normal in 10. Cystoscopy revealed normal findings in 17 cases, trabeculations in 21, inflammation in 20 and type 1 posterior urethral valves in 2. The incidence of upper tract changes and positive anatomical findings in children with voiding dysfunction is too low to justify routine radiological evaluation and cystoscopy. However, in those who present with a nonfebrile urinary tract infection there remains an important role for voiding cystourethrography. We do not recommend routine urodynamics in children with voiding disorder because this study does not change therapy or influence the final outcome. Thorough history and physical examination lead to the correct diagnosis and treatment in the majority of children. A focus on correcting faulty voiding behavior with the judicious administration of antibiotics and anticholinergic therapy leads to a favorable outcome in most cases.


Combined aclidinium bromide and long-acting beta2-agonist for chronic obstructive pulmonary disease (COPD).
Several dual bronchodilator combinations of long-acting beta2-agonist (LABA) and long-acting muscarinic antagonist (LAMA) have been approved for treatment of stable chronic obstructive pulmonary disease (COPD). The current GOLD (Global Initiative for Chronic Obstructive Lung Disease) recommendations suggest the use of LABA/LAMA combinations in people with group B COPD with persistent symptoms, group C COPD with further exacerbations on LAMA therapy alone and group D COPD with or without inhaled corticosteroids (ICS). Fixed-dose combination (FDC) of aclidinium/formoterol is one of the approved LABA/LAMA therapies for people with stable COPD. To assess the efficacy and safety of combined aclidinium bromide and long-acting beta2-agonists in stable COPD. We searched the Cochrane Airways Group Specialised Register (CAGR), ClinicalTrials.gov, World Health Organization (WHO) trials portal, United States Food and Drug Administration (FDA) and manufacturers' websites as well as the reference list of published trials up to 12 October 2018. Parallel-group randomised controlled trials (RCTs) assessing combined aclidinium bromide and LABAs in people with stable COPD. We used standard methodological procedures expected by Cochrane for data collection and analysis. The primary outcomes were exacerbations requiring a short course of an oral steroid or antibiotic, or both; quality of life measured by a validated scale and non-fatal serious adverse events (SAEs). Where the outcome or study details were not reported, we contacted the study investigators or pharmaceutical company trial co-ordinators (or both) for missing data. We identified RCTs comparing aclidinium/formoterol FDC versus aclidinium, formoterol or placebo only. We included seven multicentre trials of four to 52 weeks' duration conducted in outpatient settings. There were 5921 participants, whose mean age ranged from 60.7 to 64.7 years, mostly men with a mean smoking pack-years of 46.4 to 61.3 of which 43.9% to 63.4% were current smokers. They had a moderate-to-severe degree of COPD with a mean postbronchodilator forced expiratory volume in one second (FEV1) between 50.5% and 61% of predicted normal and the baseline mean FEV1 of 1.23 L to 1.43 L. We assessed performance and detection biases as low for all studies whereas selection, attrition and reporting biases were either low or unclear.FDC versus aclidiniumThere was no evidence of a difference between FDC and aclidinium for exacerbations requiring steroids or antibiotics, or both (OR 0.95, 95% CI 0.71 to 1.27; 2 trials, 2156 participants; moderate-certainty evidence); quality of life measured by St George's Respiratory Questionnaire (SGRQ) total score (MD -0.92, 95% CI -2.15 to 0.30); participants with significant improvement in SGRQ score (OR 1.17, 95% CI 0.97 to 1.41; 2 trials, 2002 participants; moderate-certainty evidence); non-fatal SAE (OR 1.19, 95% CI 0.79 to 1.80; 3 trials, 2473 participants; moderate-certainty evidence); hospital admissions due to severe exacerbations (OR 0.62, 95% CI 0.29 to 1.29; 2 trials, 2156 participants; moderate-certainty evidence) or adverse events (OR 0.95, 95% CI 0.76 to 1.18; 3 trials, 2473 participants; moderate-certainty evidence). Compared with aclidinium, FDC improved symptoms (Transitional Dyspnoea Index (TDI) focal score: MD 0.37, 95% CI 0.07 to 0.68; 2 trials, 2013 participants) with a higher chance of achieving a minimal clinically important difference (MCID) of at least one unit improvement (OR 1.34, 95% CI 1.11 to 1.62; high-certainty evidence); the number needed to treat for an additional beneficial outcome (NNTB) being 14 (95% CI 9 to 39).FDC versus formoterolWhen compared to formoterol, combination therapy reduced exacerbations requiring steroids or antibiotics, or both (OR 0.78, 95% CI 0.62 to 0.99; 3 trials, 2694 participants; high-certainty evidence); may decrease SGRQ total score (MD -1.88, 95% CI -3.10 to -0.65; 2 trials, 2002 participants; low-certainty evidence; MCID for SGRQ is 4 units); increased TDI focal score (MD 0.42, 95% CI 0.11 to 0.72; 2 trials, 2010 participants) with more participants attaining an MCID (OR 1.30, 95% CI 1.07 to 1.56; high-certainty evidence) and an NNTB of 16 (95% CI 10 to 60). FDC lowered the risk of adverse events compared to formoterol (OR 0.78, 95% CI 0.65 to 0.93; 5 trials, 3140 participants; high-certainty evidence; NNTB 22). However, there was no difference between FDC and formoterol for hospital admissions, all-cause mortality and non-fatal SAEs.FDC versus placeboCompared with placebo, FDC demonstrated no evidence of a difference in exacerbations requiring steroids or antibiotics, or both (OR 0.82, 95% CI 0.60 to 1.12; 2 trials, 1960 participants; moderate-certainty evidence) or hospital admissions due to severe exacerbations (OR 0.55, 95% CI 0.25 to 1.18; 2 trials, 1960 participants; moderate-certainty evidence), although estimates were uncertain. Quality of life measure by SGRQ total score was significantly better with FDC compared to placebo (MD -2.91, 95% CI -4.33 to -1.50; 2 trials, 1823 participants) resulting in a corresponding increase in SGRQ responders who achieved at least four units decrease in SGRQ total score (OR 1.72, 95% CI 1.39 to 2.13; high-certainty evidence) with an NNTB of 7 (95% CI 5 to 12). FDC also improved symptoms measured by TDI focal score (MD 1.32, 95% CI 0.96 to 1.69; 2 studies, 1832 participants) with more participants attaining at least one unit improvement in TDI focal score (OR 2.51, 95% CI 2.02 to 3.11; high-certainty evidence; NNTB 4). There were no differences in non-fatal SAEs, adverse events and all-cause mortality between FDC and placebo.Combination therapy significantly improved trough FEV1 compared to aclidinium, formoterol or placebo. FDC improved dyspnoea and lung function compared to aclidinium, formoterol or placebo, and this translated into an increase in the number of responders on combination treatment. Quality of life was better with combination compared to formoterol or placebo. There was no evidence of a difference between FDC and monotherapy or placebo for exacerbations, hospital admissions, mortality, non-fatal SAEs or adverse events. Studies reported a lower risk of moderate exacerbations and adverse events with FDC compared to formoterol; however, larger studies would yield a more precise estimate for these outcomes.


Are mammography recommendations in women younger than 40 related to increased risk?
Our objective was to examine the association between self-reported breast cancer risk factors and reported physician recommendations for mammography among women younger than 40. This study uses the 2005 National Health Interview Survey (NHIS) and includes 2,703 women ages 30-39 who reported having seen a doctor in the past 12 months. The NHIS is a population-based, cross-sectional survey of adult respondents in the United States. Overall, 19.0% of these women reported a recent mammography recommendation. Among women reporting no prior mammogram, women ages 30-34 with risk factors for breast cancer were more likely to report a recent mammogram recommendation than women without risk factors. There was no such association for women ages 35-39. Among women who reported a prior mammogram, risk factors were not associated with a recommendation for mammography; there was an association with age and recent clinical breast examination. Despite a lack of evidence-based guidelines for women under 40 years of age, these data suggest some younger women are being recommended for early mammography without indication. The relative benefits and harms of recommending mammography in this age group need further examination.


A Modified Dynamic Surface Controller for Delayed Neuromuscular Electrical Stimulation.
A widely accepted model of muscle force generation during neuromuscular electrical stimulation (NMES) is a second-order nonlinear musculoskeletal dynamics cascaded to a delayed first-order muscle activation dynamics. However, most nonlinear NMES control methods have either neglected the muscle activation dynamics or used an ad hoc strategies to tackle the muscle activation dynamics, which may not guarantee control stability. We hypothesized that a nonlinear control design that includes muscle activation dynamics can improve the control performance. In this paper, a dynamic surface control (DSC) approach was used to design a PID-based NMES controller that compensates for EMD in the activation dynamics. Because the muscle activation is unmeasurable, a model based estimator was used to estimate the muscle activation in realtime. The Lyapunov stability analysis confirmed that the newly developed controller achieves semi-global uniformly ultimately bounded (SGUUB) tracking for the musculoskeletal system. Experiments were performed on two able-bodied subjects and one spinal cord injury subject using a modified leg extension machine. These experiments illustrate the performance of the new controller and compare it to a previous PID-DC controller that did not consider muscle activation dynamics in the control design. These experiments support our hypothesis that a control design that includes muscle activation improves the NMES control performance.


Application of peritoneal dialysis in elderly patients by classifying the age into young-old, old, and oldest-old.
A greater number of end-stage renal disease patients are receiving peritoneal dialysis (PD) or hemodialysis (HD) in Japan. However, medical concerns with advancing age have been raised in PD utilization for elderly patients. The objective of this study was to address the indications for PD in elderly patients in terms of medical concerns such as nutrition state, residual renal function, dialysis efficiency, peritonitis, cardiovascular disease (CVD) complications, and technique survival. In a retrospective, two-center study, we evaluated 247 patients who newly started PD from 2002 to 2008. All patients were divided into four groups: young (<64 years, n = 99), young-old (65-74 years, n = 55), old (75-84 years, n = 62) and oldest-old (≥85 years, n = 31). Serum albumin, hemoglobin, β(2)-microglobulin, cardio-thoracic ratio, 24-hour urine collection and spent dialysate volume was collected at the initiation of PD and after 1, 2, 3, and 4 years. PD withdrawal, occurrence of CVD complications, peritonitis and death were recorded. Nephrosclerosis as a primary disease increased with advancing age (p = 0.001). At baseline, gender, body weight, serum creatinine, hemoglobin and cardio-thoracic ratio were significantly different among the four groups. No significant decrease was shown in urine output with advancing age. The spent dialysate volume was significantly lower (mean 3.8 liters/day) in the oldest-old group compared with the other groups (p = 0.001). However, a smaller volume of PD fluid in the oldest-old group was not accompanied by a significantly higher serum β(2)-microgloblin level compared with the other groups and there was no reason of PD withdrawal for underdialysis in the old and oldest-old groups. Neither the incidence of CVD complications nor that of peritonitis was increased with advancing age. There was no significant difference in technique survival rate excluding death between each group. These findings suggest that there are no medical concerns to avoid PD therapy in elderly end-stage renal disease patients.


Concepts in the prevention of adenocarcinoma of the distal esophagus and proximal stomach.
For decades, the incidence rates for squamous cell carcinoma of the esophagus and adenocarcinoma of the distal stomach have been declining while the rates for adenocarcinomas of the esophagus and gastric cardia have increased profoundly. Recent studies have shown that the gastroesophageal junction (GEJ) is regularly exposed to concentrated gastric acid and to a variety of nitrosating species, noxious agents that may contribute to carcinogenesis in this region. For adenocarcinomas that straddle the GEJ, it can be difficult to determine whether the tumor originated in the esophagus or in the gastric cardia. This classification problem hampers studies on the epidemiology and pathogenesis of GEJ tumors. Current concepts in the prevention of cancers of the distal esophagus and proximal stomach have emerged from advances in our understanding of the specific molecular events that occur during the evolution of these tumors. This report reviews those events and focuses on current concepts in the prevention of adenocarcinomas at the GEJ. The similarities and differences in risk factors, molecular pathogenesis, and in preventive strategies for adenocarcinomas of the esophagus and gastric cardia are highlighted.


Enhanced behavioral conditioning to context and impaired behavioral and neuroendocrine responses to conditioned stimuli following ceruleocortical noradrenergic lesions: support for an attentional hypothesis of central noradrenergic function.
This study tested the hypothesis that cortical noradrenaline (NA) depletion induced by 6-hydroxydopamine (6-OHDA) widens attentional span, impairing the acquisition of conditioning to an explicit stimulus while enhancing conditioning to contextual stimuli. Sham-operated and NA-depleted rats were exposed to pairings of an auditory (clicker) CS and (footshock) US in a distinctive environment. Half of the lesioned and half of the control animals were trained with a short trace interval between presentations of clicker and shock, and half with a long trace. Associative learning theory predicts that a long trace interval should bias intact animals towards stronger contextual conditioning and, in contrast, a short trace interval should bias controls towards stronger CS conditioning. During testing, NA-depleted animals showed impaired fear conditioning to explicit cues, compared with controls, indicated by their reduced suppression of drinking when the CS was introduced into a separate, lick-operant chamber. In contrast, the same animals exhibited enhanced fear of contextual cues, as shown by their greater preference for a "safe" environment over the one in which they were shocked. The behavioral and plasma corticosterone responses of individual animals to the CS were positively correlated in both the lesion and sham groups. Corticosterone levels corroborated the impairment in CS conditioning caused by the lesion. In contrast, behavioral and corticosterone responses to contextual stimuli were not significantly correlated in either group, and there was no enhancement of the corticosterone response to contextual stimuli in the lesioned animals.(ABSTRACT TRUNCATED AT 250 WORDS)


Low microwave-amplitude ESR spectroscopy: measuring spin-relaxation interactions of moderately immobilized spin labels in proteins.
Electron spin resonance (ESR) spectroscopy in combination with site-directed spin labeling (SDSL) is a powerful tool for determining protein structure, dynamics and interactions. We report here a method for determining interactions between spin labels and paramagnetic relaxation agents, which is performed under subsaturating conditions. The low microwave-field amplitude employed (h(1)<0.36 G) only requires standard, commercially available ESR equipment. The effect of relaxation enhancement on the spin-spin-relaxation time, T(2e), is measured by this method, and compared to classical progressive power saturation performed on a free spin label, (1-oxyl-2,2,5,5-tetramethyl-Delta(3)-pyrroline-3-methyl)methanethiosulfonate (MTSL), and a spin-labeled protein (Thermomyces lanuginosa lipase, TLL-I252C), employing the water-soluble relaxation agent chromium(III) oxalate (Crox) in concentrations between 0-10 mM. The low-amplitude theory showed excellent agreement with that of classical power saturation in quantifying Crox-induced relaxation enhancement. Low-amplitude measurements were then performed using a standard resonator, with Crox, on 11 spin-labeled TLL mutants displaying rotational correlation times in the motional narrowing regime. All spin-labeled proteins exhibited significant changes in T(2e). We postulate that this novel method is especially suitable for studying moderately immobilized spin labels, such as those positioned at exposed sites in a protein. This method should prove useful for research groups with access to any ESR instrumentation.


delta psi-mediated signalling in the bacteriorhodopsin-dependent photoresponse.
It has been shown previously that the proton-pumping activity of bacteriorhodopsin from Halobacterium salinarium can transmit an attractant signal to the bacterial flagella upon an increase in light intensity over a wide range of wavelengths. Here, we studied the effect of blue light on phototactic responses by the mutant strain Pho8l-B4, which lacks both sensory rhodopsins but has the ability to synthesize bacteriorhodopsin. Under conditions in which bacteriorhodopsin was largely accumulated as the M412 bacteriorhodopsin photocycle intermediate, halobacterial cells responded to blue light as a repellent. This response was pronounced when the membrane electric potential level was high in the presence of arginine, active oxygen consumption, or high-background long-wavelength light intensity but was inhibited by an uncoupler of oxidative phosphorylation (carbonyl cyanide 3-chlorophenylhydrazone) and was inverted in a background of low long-wavelength light intensity. The response to changes in the intensity of blue light under high background light was asymmetric, since removal of blue light did not produce an expected suppression of reversals. Addition of ammonium acetate, which is known to reduce the pH gradient changes across the membrane, did not inhibit the repellent effect of blue light, while the discharge of the membrane electric potential by tetraphenylphosphonium ions inhibited this sensory reaction. We conclude that the primary signal from bacteriorhodopsin to the sensory pathway involves changes in membrane potential.


Measures of Appropriateness and Value for Breast Surgeons and Their Patients: The American Society of Breast Surgeons Choosing Wisely (®) Initiative.
Current breast cancer care is based on high-level evidence from randomized, controlled trials. Despite these data, there continues to be variability of breast cancer care, including overutilization of some tests and operations. To reduce overutilization, the American Board of Internal Medicine Choosing Wisely (®) Campaign recommends that professional organizations provide patients and providers with a list of care practices that may not be necessary. Shared decision making regarding these services is encouraged. The Patient Safety and Quality Committee of the American Society of Breast Surgeons (ASBrS) solicited candidate measures for the Choosing Wisely (®) Campaign. The resulting list of "appropriateness" measures of care was ranked by a modified Delphi appropriateness methodology. The highest-ranked measures were submitted to and later approved by the ASBrS Board of Directors. They are listed below. (1) Don't routinely order breast magnetic resonance imaging in new breast cancer patients. (2) Don't routinely excise all the lymph nodes beneath the arm in patients having lumpectomy for breast cancer. (3) Don't routinely order specialized tumor gene testing in all new breast cancer patients. (4) Don't routinely reoperate on patients with invasive cancer if the cancer is close to the edge of the excised lumpectomy tissue. (5) Don't routinely perform a double mastectomy in patients who have a single breast with cancer. The ASBrS list for the Choosing Wisely (®) campaign is easily accessible to breast cancer patients online. These measures provide surgeons and their patients with a starting point for shared decision making regarding potentially unnecessary testing and operations.


Structure-function Evaluation of Stem Cell Therapies for Spinal Cord Injury.
Spinal cord injuries (SCI) are prevalent, devastating for quality and expectancy of life, and cause heavy economic burdens. Stem cell therapies hold promise in complete structural and functional restoration of SCI. This review focuses on the methods currently used to evaluate the stem cell therapies for SCI. Various kinds of stem cells involving embryonic stem cells (ESCs), bone marrow stromal cells (BMSCs), neural stem cells (NSCs) and induced pluripotent stem cells (iPSCs) are extensively used in regenerative research of SCI. For evaluation, the survival and integration of transplanted cells, spinal cord reconstruction and functional recovery all should be considered. Histological and histochemistrical, microscopic, and colorimetric assays, and real-time RT-PCR techniques are applied to determine the outcome. From the three main aspects-transplanted cells, spinal cord structure, and functional recovery-we summarize and discuss these methods with certain instances of applications in SCI models. Importantly, for the evaluations of function, neuronal transmitting, electrophysiological analysis and behavioral score are included. Wider conjunction of established technologies, as well as the further development of nondestructive methods might make a big difference in testing stem cell therapies.


Management of venous thromboembolism in patients with advanced cancer: a systematic review and meta-analysis.
Venous thromboembolism is common in patients with cancer. However, no management guidelines exist for venous thromboembolism specific to patients with advanced progressive cancer. To help develop recommendations for practice, we have done a comprehensive review of anticoagulation treatment in patients with cancer, with particular focus on studies that included patients with advanced disease. Data from 19 publications, including randomised, prospective, and retrospective studies suggest that: long-term full-dose low-molecular-weight heparin (LMWH) is more effective than warfarin in the secondary prophylaxis of venous thromboembolism in patients with cancer of any stage, performance status, or prognosis; warfarin should not be used in patients with advancing progressive disease; and in patients at high risk of bleeding, full-dose LMWH for 7 days followed by a long-term decreased fixed dose long term can be considered. The optimum treatment duration is unclear, but because the prothrombotic tendency will persist in patients with advanced cancer, indefinite treatment is generally recommended. For patients with contraindications to anticoagulation, inferior-vena-caval filters can be considered, but their use needs careful patient selection. Ultimately, the decision to initiate, continue, and stop anticoagulation will need to be made on an individual basis, guided by the available evidence, the patient's circumstances, and their informed preferences.


Observer variation in visual assessment of forelimb horseshoe characteristics on Thoroughbred racehorses.
To assess the accuracy and reliability of a visual method of evaluating horseshoe characteristics. 1,199 Thoroughbred racehorses. Characteristics of 1 forelimb horseshoe were visually assessed on horses immediately prior to racing by 5 field observers at 5 major racetracks in California. Characteristics evaluated included horseshoe type; toe grab height; and the presence of a rim, pad, and heel traction devices. Sensitivity and specificity for observer assessment of horseshoe characteristics were calculated by comparing observer assessments to a postmortem laboratory standard for horses that died within 48 hours of a race. Intraobserver agreement was assessed in a subset of horses by comparing horseshoe observations made before and after the horse's race. Interobserver agreement was evaluated by comparing horseshoe assessment among observers who examined the same subset of horses prior to racing on select days. The sensitivity and specificity of this visual method of evaluating horseshoe characteristics were good and ranged from 0.75 to 1 and 0.67 to 1, respectively. Agreement beyond chance (weighted kappa values) between observers and the laboratory standard for toe grab height was fair (0.60 to 0.62). Intraobserver and interobserver agreements (kappa values) were high (0.86 to 0.99 and 0.71 to 1, respectively). Visual observation of horseshoes can be a feasible and reproducible method for assessing horseshoe characteristics prospectively in a large cohort of horses under racing conditions.


Pyruvate dehydrogenase complex stimulation promotes immunometabolic homeostasis and sepsis survival.
Limited understanding of the mechanisms responsible for life-threatening organ and immune failure hampers scientists' ability to design sepsis treatments. Pyruvate dehydrogenase kinase 1 (PDK1) is persistently expressed in immune-tolerant monocytes of septic mice and humans and deactivates mitochondrial pyruvate dehydrogenase complex (PDC), the gate-keeping enzyme for glucose oxidation. Here, we show that targeting PDK with its prototypic inhibitor dichloroacetate (DCA) reactivates PDC; increases mitochondrial oxidative bioenergetics in isolated hepatocytes and splenocytes; promotes vascular, immune, and organ homeostasis; accelerates bacterial clearance; and increases survival. These results indicate that the PDC/PDK axis is a druggable mitochondrial target for promoting immunometabolic and organ homeostasis during sepsis.


Public reactions to e-cigarette regulations on Twitter: a text mining analysis.
In May 2016, the Food and Drug Administration (FDA) issued a final rule that deemed e-cigarettes to be within their regulatory authority as a tobacco product. News and opinions about the regulation were shared on social media platforms, such as Twitter, which can play an important role in shaping the public's attitudes. We analysed information shared on Twitter for insights into initial public reactions. A text mining approach was used to uncover important topics among reactions to the e-cigarette regulations on Twitter. SAS Text Miner V.12.1 software was used for descriptive text mining to uncover the primary topics from tweets collected from May 1 to May 17 2016 using NUVI software to gather the data. A total of nine topics were generated. These topics reveal initial reactions to whether the FDA's e-cigarette regulations will benefit or harm public health, how the regulations will impact the emerging e-cigarette market and efforts to share the news. The topics were dominated by negative or mixed reactions. In the days following the FDA's announcement of the new deeming regulations, the public reaction on Twitter was largely negative. Public health advocates should consider using social media outlets to better communicate the policy's intentions, reach and potential impact for public good to create a more balanced conversation.


Managing anxiety in eating disorders with knitting.
Recovery from anorexia nervosa (AN) is often confounded by intrusive, anxious preoccupations with control of eating, weight and shape. These are distressing and represent a potential barrier to psychological change. Theoretical and empirical evidence suggests that performing a concurrent visuospatial task reduces the emotional intensity of distressing images. We assessed whether the visuospatial task of knitting influences the anxious preoccupation experienced by inpatients with AN. Prospective interventional cohort. Thirty-eight women with AN admitted to a specialized eating disorder unit. All subjects were given knitting lessons and free access to supplies. Subjects were asked to report the qualitative effects of knitting on their psychological state using a self-report questionnaire. Patients reported a subjective reduction in anxious preoccupation when knitting. In particular, 28/38 (74%) reported it lessened the intensity of their fears and thoughts and cleared their minds of eating disorder preoccupations, 28/38 (74%) reported it had a calming and therapeutic effect and 20/38 (53%) reported it provided satisfaction, pride and a sense of accomplishment. This preliminary data suggests that knitting may benefit inpatients with eating disorders by reducing their anxious preoccupations about eating, weight and shape control. The specificity of this effect is yet to be determined. This preliminary outcome requires further controlled study in AN subjects. From a clinical perspective, knitting is inexpensive, easily learned, can continue during social interaction, and can provide a sense of accomplishment. The theoretical and empirical rationale for this observation, and implications for deriving alternative strategies to augment treatment in AN, are discussed.


Renal function is a determinant of subjective well-being in active seniors but not in patients with subjective memory complaints.
During our whole life span, factors influencing health and functioning are accumulated. In chronic kidney disease, quality of life is adversely affected. We hypothesized that biomarkers of renal function could also be determinants of subjective well-being (SWB) in Swedish elderly subjects. SWB was assessed by the Psychological General Well-Being index (PGWB index) in two study groups: Active seniors (AS) consisted of community-dwelling elderly Swedes leading an active life (n = 389), and the DGM cohort (n = 300) consisted of subjects referred to the Memory Unit at the Department of Geriatrics for memory problems, Serum creatinine, cystatin C, and eGFR (CKD-EPI) were used as biomarkers of renal function. There were no significant differences in cystatin C and eGFR values between the two cohorts: cystatin C medians 0.88 vs 0.86 mg/L and eGFR 73 vs 80 mL/min/1.73 m2 (AS vs DGM). In the AS cohort cystatin C was negatively related to PGWB index in women (P < 0.001, R2 ≈ 5%), and the covariates age and BMI did not improve the models. The renal biomarkers were unrelated to the PGWB index in the DGM cohort. Cystatin C in the AS cohort was adversely related to the PGWB subdimensions anxiety, depressed mood, positive well-being, and vitality in women, but in men only to depressed mood (P < 0.006; R2 ≈ 6%). In the DGM cohort, depressed mood in men was also significantly related to cystatin C (P = 0.050), but not in women. Renal function even within the normal range, measured by serum cystatin C concentration, has significant and sex specific associations with subjective well-being and its subdimensions in healthy elderly subjects. Maintenance of good renal function in aging may be of importance in maintaining a high subjective well-being.


Estimation and Inference of Quantile Regression for Survival Data Under Biased Sampling.
Biased sampling occurs frequently in economics, epidemiology, and medical studies either by design or due to data collecting mechanism. Failing to take into account the sampling bias usually leads to incorrect inference. We propose a unified estimation procedure and a computationally fast resampling method to make statistical inference for quantile regression with survival data under general biased sampling schemes, including but not limited to the length-biased sampling, the case-cohort design, and variants thereof. We establish the uniform consistency and weak convergence of the proposed estimator as a process of the quantile level. We also investigate more efficient estimation using the generalized method of moments and derive the asymptotic normality. We further propose a new resampling method for inference, which differs from alternative procedures in that it does not require to repeatedly solve estimating equations. It is proved that the resampling method consistently estimates the asymptotic covariance matrix. The unified framework proposed in this article provides researchers and practitioners a convenient tool for analyzing data collected from various designs. Simulation studies and applications to real datasets are presented for illustration. Supplementary materials for this article are available online.


Investigating the neural correlates of smoking: Feasibility and results of combining electronic cigarettes with fMRI.
Cigarette addiction is driven partly by the physiological effects of nicotine, but also by the distinctive sensory and behavioural aspects of smoking, and understanding the neural effects of such processes is vital. There are many practical difficulties associated with subjects smoking in the modern neuroscientific laboratory environment, however electronic cigarettes obviate many of these issues, and provide a close simulation of smoking tobacco cigarettes. We have examined the neural effects of 'smoking' electronic cigarettes with concurrent functional Magnetic Resonance Imaging (fMRI). The results demonstrate the feasibility of using these devices in the MRI environment, and show brain activation in a network of cortical (motor cortex, insula, cingulate, amygdala) and sub-cortical (putamen, thalamus, globus pallidus, cerebellum) regions. Concomitant relative deactivations were seen in the ventral striatum and orbitofrontal cortex. These results reveal the brain processes involved in (simulated) smoking for the first time, and validate a novel approach to the study of smoking, and addiction more generally.


Robot visions.
This article explores the resonating figures of primate, child, and robot in contemporary technoscientific corporealizations of the 'almost human'. We take as our model (in)organism 'Lucy the Robot Orangutan', roboticist Steve Grand's project to create an artificial life form with a mind of its own. One aspect of Lucy's figuration by Grand, we argue, which ties her to Haraway's analysis of the primate, is of the robot as a model for animal, and more specifically (or aspirationally) human, cognition. We follow the trope of 'model organism' as it is under discussion within science and technology studies and as an ironic descriptor for our own interest in Lucy as an entity/project through which to illuminate figurations within robotics more widely. Primate and robot together are forms of natureculture that help to clarify how the categories of animal and machine are entangled, while making explicit investments in their differences from one another, and from the third category of the human. We conclude, again following Haraway, by imagining what other possibilities there might be for figuring humans, robots, and their relations if we escape the reiterative imaginary of the robot as proxy for becoming human.


Interventions to improve osteoporosis treatment following hip fracture. A prospective, randomized trial.
Treatment of osteoporosis following a hip fracture has been notoriously poor. Many efforts have been made to improve treatment rates. The purpose of this study was to determine whether a perioperative inpatient intervention program, involving patient education and providing a list of questions for the primary care physician, increased the percentage of patients in whom osteoporosis was addressed following a hip fracture. A prospective, randomized trial involving eighty patients who had been admitted to an academic medical center with a low-energy hip fracture was conducted. During their hospitalization, the study group patients were engaged in a fifteen-minute discussion regarding the association between osteoporosis and hip fractures, the efficacy of dual-energy x-ray absorptiometry scans in the diagnosis of osteoporosis and of bisphosphonates in its treatment, and the importance of medical follow-up for osteoporosis management. These patients were also provided with five questions regarding osteoporosis treatment to be given to their primary medical physician, and they were reminded about the questions during a follow-up telephone call six weeks later. The patients in the control group received a brochure describing methods for preventing falls. Both groups were contacted by telephone at six months after discharge to determine whether osteoporosis had been addressed. Positive indicators of intervention included assessment of bone mineral density with dual-energy x-ray absorptiometry and initiation of antiresorptive therapy. The average age in each group was eighty-two years, and 78% of the patients were female. Four patients in each group did not survive through the six-month follow-up period and were excluded from the trial. Fifteen (42%) of the thirty-six patients who had been randomized to the study group, compared with only seven (19%) of the thirty-six patients in the control group, had their osteoporosis addressed by their primary physician. This difference between the groups was significant (p = 0.036). Patients who were provided with information and questions for their primary care physician about osteoporosis were more likely to receive appropriate therapeutic intervention than were patients who had not received the information and questions. Orthopaedic surgeons have a unique opportunity to improve the rate of osteoporosis treatment in the perioperative period following a hip fracture by educating patients and directing them toward channels for long-term osteoporosis management.


Evolutionary changes of multiple visual pigment genes in the complete genome of Pacific bluefin tuna.
Tunas are migratory fishes in offshore habitats and top predators with unique features. Despite their ecological importance and high market values, the open-ocean lifestyle of tuna, in which effective sensing systems such as color vision are required for capture of prey, has been poorly understood. To elucidate the genetic and evolutionary basis of optic adaptation of tuna, we determined the genome sequence of the Pacific bluefin tuna (Thunnus orientalis), using next-generation sequencing technology. A total of 26,433 protein-coding genes were predicted from 16,802 assembled scaffolds. From these, we identified five common fish visual pigment genes: red-sensitive (middle/long-wavelength sensitive; M/LWS), UV-sensitive (short-wavelength sensitive 1; SWS1), blue-sensitive (SWS2), rhodopsin (RH1), and green-sensitive (RH2) opsin genes. Sequence comparison revealed that tuna's RH1 gene has an amino acid substitution that causes a short-wave shift in the absorption spectrum (i.e., blue shift). Pacific bluefin tuna has at least five RH2 paralogs, the most among studied fishes; four of the proteins encoded may be tuned to blue light at the amino acid level. Moreover, phylogenetic analysis suggested that gene conversions have occurred in each of the SWS2 and RH2 loci in a short period. Thus, Pacific bluefin tuna has undergone evolutionary changes in three genes (RH1, RH2, and SWS2), which may have contributed to detecting blue-green contrast and measuring the distance to prey in the blue-pelagic ocean. These findings provide basic information on behavioral traits of predatory fish and, thereby, could help to improve the technology to culture such fish in captivity for resource management.


The transcription factor EPAS1 links DOCK8 deficiency to atopic skin inflammation via IL-31 induction.
Mutations of DOCK8 in humans cause a combined immunodeficiency characterized by atopic dermatitis with high serum IgE levels. However, the molecular link between DOCK8 deficiency and atopic skin inflammation is unknown. Here we show that CD4+ T cells from DOCK8-deficient mice produce large amounts of IL-31, a major pruritogen associated with atopic dermatitis. IL-31 induction critically depends on the transcription factor EPAS1, and its conditional deletion in CD4+ T cells abrogates skin disease development in DOCK8-deficient mice. Although EPAS1 is known to form a complex with aryl hydrocarbon receptor nuclear translocator (ARNT) and control hypoxic responses, EPAS1-mediated Il31 promoter activation is independent of ARNT, but in collaboration with SP1. On the other hand, we find that DOCK8 is an adaptor and negative regulator of nuclear translocation of EPAS1. Thus, EPAS1 links DOCK8 deficiency to atopic skin inflammation via IL-31 induction in CD4+ T cells.


Opioid-Induced Adrenal Insufficiency.
One in 10 Americans experience chronic pain. Although opioids do play a role in the management of pain, long-term opioid use may lead to adverse effects. Endocrine-related adverse effects have been described but remain poorly recognized. Opioid-induced adrenal insufficiency occurs because of suppression of hypothalamic-pituitary-adrenal communication and may be challenging to diagnose but has been reported in 9% to 29% of patients receiving long-term opiate therapy. Little data exist to guide case detection and patient management. Treatment includes cessation of opiates (the inciting factor) if possible and glucocorticoid replacement.


Biolistic transfection of neurons.
One method used to study gene function is through the manipulation of gene expression by transfecting cells with DNA constructs designed to overexpress or knock out particular proteins. Unfortunately, transfection of cells and tissues remains a rate-limiting step for molecular studies in many fields, especially neurobiology. Conventional transfection techniques are of limited effectiveness, particularly in intact tissue. This protocol describes an alternative method for transfecting cells, called biolistics. Biolistics is a physical method of transfection in which target tissue is bombarded with DNA-coated gold particles using a "gene gun," produced by Bio-Rad Laboratories. Cells penetrated by gold particles have a high likelihood of becoming transfected. Because biolistic transfection relies only on the physical penetration of a cell's membrane, it is possible to use biolistics to transfect cells that are resistant to transfection by other methods, such as neurons in primary culture and organotypic slice cultures. This protocol provides information on optimizing the biolistic parameters for transfecting neurons in both of these preparations. Once optimized, biolistic transfection is a reliable and efficient method for studying gene function in many cell types, especially postmitotic neurons.


Computational multiscale toxicodynamic modeling of silver and carbon nanoparticle effects on mouse lung function.
A computational, multiscale toxicodynamic model has been developed to quantify and predict pulmonary effects due to uptake of engineered nanomaterials (ENMs) in mice. The model consists of a collection of coupled toxicodynamic modules, that were independently developed and tested using information obtained from the literature. The modules were developed to describe the dynamics of tissue with explicit focus on the cells and the surfactant chemicals that regulate the process of breathing, as well as the response of the pulmonary system to xenobiotics. Alveolar type I and type II cells, and alveolar macrophages were included in the model, along with surfactant phospholipids and surfactant proteins, to account for processes occurring at multiple biological scales, coupling cellular and surfactant dynamics affected by nanoparticle exposure, and linking the effects to tissue-level lung function changes. Nanoparticle properties such as size, surface chemistry, and zeta potential were explicitly considered in modeling the interactions of these particles with biological media. The model predictions were compared with in vivo lung function response measurements in mice and analysis of mice lung lavage fluid following exposures to silver and carbon nanoparticles. The predictions were found to follow the trends of observed changes in mouse surfactant composition over 7 days post dosing, and are in good agreement with the observed changes in mouse lung function over the same period of time.


Nickel complexes of a pincer amidobis(amine) ligand: synthesis, structure, and activity in stoichiometric and catalytic C-C bond-forming reactions of alkyl halides.
The synthesis, properties, and reactivity of nickel(II) complexes of a newly developed pincer amidobis(amine) ligand ((Me)NN(2)) are described. Neutral or cationic complexes [((Me)NN(2))NiX] (X = OTf (6), OC(O)CH(3) (7), CH(3)CN (8), OMe (9)) were prepared by salt metathesis or chloride abstraction from the previously reported [((Me)NN(2))NiCl] (1). The Lewis acidity of the {((Me)NN(2))Ni} fragment was measured by the (1)H NMR chemical shift of the coordinated CH(3)CN molecule in 8. Electrochemical measurements on 1 and 8 indicate that the electron-donating properties of NN(2) are similar to those of the analogous amidobis(phosphine) (pnp) ligands. The solid-state structures of 6-8 were determined and compared to those of 1 and [((Me)NN(2))NiEt] (3). In all complexes, the (Me)NN(2) ligand coordinates to the Ni(II) ion in a mer fashion, and the square-planar coordination sphere of the metal is completed by an additional donor. The coordination chemistry of (Me)NN(2) thus resembles that of other three-dentate pincer ligands, for example, pnp and arylbis(amine) (ncn). Reactions of 2 with alkyl monohalides, dichlorides, and trichlorides were investigated. Selective C-C bond formation was observed in many cases. Based on these reactions, efficient Kumada-Corriu-Tamao coupling of unactivated alkyl halides and alkyl Grignard reagents with 1 as the precatalyst was developed. Good yields were obtained for the coupling of primary and secondary iodides and bromides. Double C-C coupling of CH(2)Cl(2) with alkyl Grignard reagents by 1 was also realized. The scope and limitations of these transformations were studied. Evidence was found for a radical pathway in Ni-catalyzed C-C cross-coupling reactions, which involves Ni(II) alkyl intermediates.


Exploring the therapeutic power of narrative at the end of life: a qualitative analysis of narratives emerging in dignity therapy.
To understand the therapeutic effect of a narrative intervention, specifically dignity therapy, in patients at the end-of-life. To examine the thematic dimensions and shared narrative features of the stories that emerge in dignity therapy and theorise their relationship to the intervention's clinical impact. Resident physicians, as part of an educational intervention, co-administered the dignity therapy protocol with the principal investigator. Interviews were transcribed, edited, and then, within a week, read back to the patient and provided as a document for the patient to keep. A constant comparative approach was taken to identify narratives and thematic patterns. 12 Patients at the end-of-life were administered dignity interviews by 12 resident physicians, accompanied by the principal investigator. Palliative care settings in two University of Toronto academic hospitals. Three narrative types emerged, each containing several themes. Evaluation narratives create a life lived before illness, with an overarching theme of overcoming adversity. Transition narratives describe a changing health situation and its meanings, including impact on family and on one's world view. Legacy narratives discuss the future without the patient and contain the parables and messages to be left for loved ones. While the interview protocol guides patients' responses, the commonality of narrative structures across interviews suggests that patients draw on experiences with two familiar genres: the eulogy and the medical interview, to create a narrative order during the chaos of dying. The dignity interview's resonance with these genres appears to facilitate a powerful, and perhaps unexpected sense of agency.


Reclamation of Real Urban Wastewater Using Solar Advanced Oxidation Processes: An Assessment of Microbial Pathogens and 74 Organic Microcontaminants Uptake in Lettuce and Radish.
In this study, disinfection of urban wastewater (UWW) with two solar processes (H2O2 -20 mg/L and photo-Fenton 10 mg/L-Fe2+/20 mg/L-H2O2 at natural water pH) at pilot scale using a 60 L compound parabolic collector reactor for irrigation of two raw-eaten vegetables (lettuce and radish) has been investigated. Several microbial targets (total coliforms, Escherichia coli, Salmonella spp., and Enterococcus spp.) naturally occurring in UWW and 74 organic microcontaminants (OMCs) were monitored. Disinfection results showed no significant differences between both processes, showing the following inactivation resistance order: Salmonella spp. < E. coli < total coliforms < Enterococcus spp. Reductions of target microorganisms to concentrations below the limit of detection (LOD) was achieved in all cases with cumulative solar UV energy per volume (QUV) ranged from 12 to 40 kJ/L (90 min to 5 h). Solar photo-Fenton showed a reduction of 66% of OMCs and solar/H2O2 of 56% in 5 h treatment. Irrigation of radish and lettuce with solar treated effluents, secondary effluents, and mineral water was performed for 6 and 16 weeks, respectively. The presence of bacteria was monitored in surfaces and uptake of leaves, fruit, and also in soil. The bacterial concentrations detected were below the LOD in the 81.2% (lettuce) and the 87.5% (radish) of the total number of samples evaluated. Moreover, uptake of OMCs was reduced above 70% in crops irrigated with solar treated effluents in comparison with secondary effluents of UWW.