Communicative function demonstration induces kind-based artifact representation in preverbal infants.
Human infants grow up in environments populated by artifacts. In order to acquire knowledge about different kinds of human-made objects, children have to be able to focus on the information that is most relevant for sorting artifacts into categories. Traditional theories emphasize the role of superficial, perceptual features in object categorization. In the case of artifacts, however, it is possible that abstract, non-obvious properties, like functions, may form the basis of artifact kind representations from an early age. Using an object individuation paradigm we addressed the question whether non-verbal communicative demonstration of the functional use of artifacts makes young infants represent such objects in terms of their kinds. When two different functions were sequentially demonstrated on two novel objects as they emerged one-by-one from behind a screen, 10-month-old infants inferred the presence of two objects behind the occluder. We further show that both the presence of communicative signals and causal intervention are necessary for 10-month-olds to generate such a numerical expectation. We also found that communicative demonstration of two different functions of a single artifact generated the illusion of the presence of two objects. This suggests that information on artifact function was used as an indicator of kind membership, and infants expected one specific function to define one specific artifact kind. Thus, contrary to previous accounts, preverbal infants' specific sensitivity to object function underlies, guides, and supports their learning about artifacts.


Ocimum sanctum induces apoptosis in A549 lung cancer cells and suppresses the in vivo growth of Lewis lung carcinoma cells.
Although Ocimum sanctum has been used extensively for its medicinal values in India and China, its antitumor activity against human nonsmall cell lung carcinoma (NSCLC) A549 cells has not been investigated until now. Therefore, the antitumor mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated in A549 cells in vitro and the Lewis lung carcinoma (LLC) animal model. EEOS exerted cytotoxicity against A549 cells, increased the sub-G1 population and exhibited apoptotic bodies in A549 cells. Furthermore, EEOS cleaved poly(ADP-ribose)polymerase (PARP), released cytochrome C into cytosol and simultaneously activated caspase-9 and -3 proteins. Also, EEOS increased the ratio of proapoptotic protein Bax/antiapoptotic protein Bcl-2 and inhibited the phosphorylation of Akt and extracellular signal regulated kinase (ERK) in A549 cancer cells. In addition, it was found that EEOS can suppress the growth of LLC inoculated onto C57BL/6 mice in a dose-dependent manner. Overall, these results demonstrate that EEOS induces apoptosis in A549 cells via a mitochondria caspase dependent pathway and inhibits the in vivo growth of LLC, suggesting that EEOS can be applied to lung carcinoma as a chemopreventive candidate.


The effect of age on fluid intelligence is fully mediated by physical health.
The present study investigated the extent to which the effect of age on cognitive ability is predicted by individual differences in physical health. The sample consisted of 118 volunteer subjects who were healthy and ranging in age from 26 to 91. The examinations included a clinical investigation, magnetic resonance imaging (MRI) brain neuroimaging, and a comprehensive neuropsychological assessment. The effect of age on fluid IQ with and without visual spatial praxis and on crystallized IQ was tested whether being fully-, partially- or non-mediated by physical health. Structural equation analyses showed that the best and most parsimonious fit to the data was provided by models that were fully mediated for fluid IQ without praxis, non-mediated for crystallized IQ and partially mediated for fluid IQ with praxis. The diseases of the circulatory and nervous systems were the major mediators. It was concluded from the pattern of findings that the effect of age on fluid intelligence is fully mediated by physical health, while crystallized intelligence is non-mediated and visual spatial praxis is partially mediated, influenced mainly by direct effects of age. Our findings imply that improving health by acting against the common age-related circulatory- and nervous system diseases and risk factors will oppose the decline in fluid intelligence with age.


Epigenetic stability, adaptability, and reversibility in human embryonic stem cells.
The stability of human embryonic stem cells (hESCs) is of critical importance for both experimental and clinical applications. We find that as an initial response to altered culture conditions, hESCs change their transcription profile for hundreds of genes and their DNA methylation profiles for several genes outside the core pluripotency network. After adaption to conditions of feeder-free defined and/or xeno-free culture systems, expression and DNA methylation profiles are quite stable for additional passaging. However, upon reversion to the original feeder-based culture conditions, numerous transcription changes are not reversible. Similarly, although the majority of DNA methylation changes are reversible, highlighting the plasticity of DNA methylation, a few are persistent. Collectively, this indicates these cells harbor a memory of culture history. For culture-induced DNA methylation changes, we also note an intriguing correlation: hypomethylation of regions 500-2440 bp upstream of promoters correlates with decreased expression, opposite to that commonly seen at promoter-proximal regions. Lastly, changes in regulation of G-coupled protein receptor pathways provide a partial explanation for many of the unique transcriptional changes observed during hESC adaptation and reverse adaptation.


Severity and properties of cardiac damage caused by Streptococcus pneumoniae are strain dependent.
Streptococcus pneumoniae is an opportunistic Gram-positive pathogen that can cause invasive disease. Recent studies have shown that S. pneumoniae is able to invade the myocardium and kill cardiomyocytes, with one-in-five adults hospitalized for pneumococcal pneumonia having a pneumonia-associated adverse cardiac event. Furthermore, clinical reports have shown up to a 10-year increased risk of adverse cardiac events in patients formerly hospitalized for pneumococcal bacteremia. In this study, we investigated the ability of nine S. pneumoniae clinical isolates, representing eight unique serotypes, to cause cardiac damage in a mouse model of invasive disease. Following intraperitoneal challenge of C57BL/6 mice, four of these strains (D39, WU2, TIGR4, and 6A-10) caused high-grade bacteremia, while CDC7F:2617-97 and AMQ16 caused mid- and low-grade bacteremia, respectively. Three strains did not cause any discernible disease. Of note, only the strains capable of high-grade bacteremia caused cardiac damage, as inferred by serum levels of cardiac troponin-I. This link between bacteremia and heart damage was further corroborated by Hematoxylin & Eosin and Trichrome staining which showed cardiac cytotoxicity only in D39, WU2, TIGR4, and 6A-10 infected mice. Finally, hearts infected with these strains showed varying histopathological characteristics, such as differential lesion formation and myocytolysis, suggesting that the mechanism of heart damage varied between strains.


Oils and cancer.
Epidemiologic evidence on the relationship between mineral oil exposure and cancer is reviewed. The review is restricted to occupations involving substantial dermal and inhalational exposure and for which an epidemiologic literature exists: metal machining, print press operating, and cotton and jute spinning. Mineral oils are complex mixtures of aliphatic hydrocarbons, naphthenics, and aromatics, the relative distribution of which depends on the source of the oil and the method of refinement. End-use products contain a variety of additives, and contamination by other agents generally occurs during use. Suspect agents include polycyclic aromatic hydrocarbons (PAH) (particularly benz[a]pyrene), nitrosamines, chlorinated paraffins, long-chain aliphatics, sulfur, N-phenyl-2-naphthylamine, and formaldehyde. The heterogeneity of this exposure makes epidemiologic study difficult and meta-analysis inappropriate. Nonetheless, several associations emerge from the literature with varying degrees of support. There is clear evidence that early formulations of mineral oils used in cotton and jute spinning and in metal machining were carcinogenic to the skin. Associations of mineral oil exposure with laryngeal and rectal cancer have received some support in the literature, particularly with respect to straight oils. Evidence is suggestive that grinding operations (which can entail either mineral oil-based or ethanolamine-based fluids) are associated with excess risk of cancer of the esophagus, stomach, and pancreas. A number of bladder cancer case-control studies have noted an association with work as a machinist. There is limited evidence of an association with cancer of the colon, prostate, and sinonasal region. Several studies of printers have yielded positive findings for lung cancer, whereas studies in metal machinists have been generally negative. The PAH and nitrosamine content of current formulations is lower than in the past and the implications of these changes in composition to the carcinogenicity of the formulations are not yet known.


Dipeptidylpeptidase 4 negatively regulates colony-stimulating factor activity and stress hematopoiesis.
Enhancement of hematopoietic recovery after radiation, chemotherapy, or hematopoietic stem cell (HSC) transplantation is clinically relevant. Dipeptidylpeptidase (DPP4) cleaves a wide variety of substrates, including the chemokine stromal cell-derived factor-1 (SDF-1). In the course of experiments showing that inhibition of DPP4 enhances SDF-1-mediated progenitor cell survival, ex vivo cytokine expansion and replating frequency, we unexpectedly found that DPP4 has a more general role in regulating colony-stimulating factor (CSF) activity. DPP4 cleaved within the N-termini of the CSFs granulocyte-macrophage (GM)-CSF, G-CSF, interleukin-3 (IL-3) and erythropoietin and decreased their activity. Dpp4 knockout or DPP4 inhibition enhanced CSF activities both in vitro and in vivo. The reduced activity of DPP4-truncated versus full-length human GM-CSF was mechanistically linked to effects on receptor-binding affinity, induction of GM-CSF receptor oligomerization and signaling capacity. Hematopoiesis in mice after radiation or chemotherapy was enhanced in Dpp4(-/-) mice or mice receiving an orally active DPP4 inhibitor. DPP4 inhibition enhanced engraftment in mice without compromising HSC function, suggesting the potential clinical utility of this approach.


Impact of histologic subtype on cancer-specific survival in patients with renal cell carcinoma and tumor thrombus.
Although different prognostic factors for patients with renal cell carcinoma (RCC) and vena cava tumor thrombus (TT) have been studied, the prognostic value of histologic subtype in these patients remains unclear. We analyzed the impact of histologic subtype on cancer-specific survival (CSS). We retrospectively analyzed the records of 1774 patients with RCC and TT who underwent radical nephrectomy and tumor thrombectomy from 1971 to 2012 at 22 US and European centers. Multivariable ordered logistic and Cox regression models were used to quantify the impact of tumor histology on CSS. Overall 5-yr CSS was 53.4% (confidence interval [CI], 50.5-56.2) in the entire group. TT level (according to the Mayo classification of macroscopic venous invasion in RCC) was I in 38.5% of patients, II in 30.6%, III in 17.3%, and IV in 13.5%. Histologic subtypes were clear cell renal cell carcinoma (cRCC) in 89.9% of patients, papillary renal cell carcinoma (pRCC) in 8.5%, and chromophobe RCC in 1.6%. In univariable analysis, pRCC was associated with a significantly worse CSS (p<0.001) compared with cRCC. In multivariable analysis, the presence of pRCC was independently associated with CSS (hazard ratio: 1.62; CI, 1.01-2.61; p<0.05). Higher TT level, positive lymph node status, distant metastasis, and fat invasion were also independently associated with CSS. In our multi-institutional series, we found that patients with pRCC and vena cava TT who underwent radical nephrectomy and tumor thrombectomy had significantly worse cancer-specific outcomes when compared with patients with other histologic subtypes of RCC. We confirmed that higher TT level and fat invasion were independently associated with reduced CSS.


Blood pressure is associated with the presence and severity of nonalcoholic fatty liver disease across the spectrum of cardiometabolic risk.
To determine the relationship between clinically relevant blood pressure (BP) groups and nonalcoholic fatty liver disease (NAFLD) presence and severity especially in the milieu of other metabolic risk factors. From a Brazilian cohort of 5362 healthy middle-aged men and women who presented for yearly physical examination and testing, the cross-sectional relationship between BP categories and NAFLD was assessed. BP groups were categorized as normal, prehypertension (PHT), and hypertension (HTN) according to the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure classification. NAFLD was ultrasound diagnosed, excluding persons with alcohol consumption more than 20 g/day. NAFLD severity was estimated using the Fibrosis-4 (FIB-4) risk score. The prevalence of NAFLD was 36.2%. Participants with NAFLD were older (mean 46 vs. 42 years, P < 0.001) and had elevated BMI (mean 29.0 vs. 24.7 kg/m, P < 0.001). The prevalence of NAFLD among persons with normal BP, PHT, and HTN was 16.5, 37.5, and 59.3%, respectively. In multivariate analyses, PHT and HTN were associated with elevated odds of NAFLD (PHT-adjusted odds ratio 1.3, 95% confidence interval 1.1, 1.6; HTN-adjusted odds ratio 1.8, 95% confidence interval 1.4-2.3) compared with normal BP. Among nonobese hypertensive patients, BP control (BP < 140/90 mmHg) was independently associated with 40% lower odds of prevalent NAFLD. Compared with hypertensive patients, both normotensive individuals and prehypertensive patients were more likely to have a low fibrosis risk (FIB-4 ≥ 1.3). Prevalent NAFLD may be seen early in the development of hypertension, even in the absence of other metabolic risk factors. Controlling BP among nonobese hypertensive patients may be beneficial in preventing or limiting NAFLD.


Survival of Teliospores of Tilletia indica in Soil.
This study was conducted to assess survival of Tilletia indica teliospores in a location in the northern United States. Soils differing in texture and other characteristics were collected from four locations, equilibrated to -0.3 MPa, and infested with teliospores of T. indica to give a density of 103 teliospores per gram of dry soil. Samples (22 g) of the infested soil were placed in 20-μm mesh polyester bags, which were sealed and placed at 2-, 10-, and 25-cm depths in polyvinyl chloride tubes containing the same field soil as the infested bags. Tubes were buried vertically in the ground at Bozeman, MT, in October 1997. Soil samples were assayed for recovery and germination of T. indica teliospores 1 day and 8, 20, and 32 months after incorporation of teliospores into soil. The rates of teliospores recovered from soil samples were 90.2, 18.7, 16.1, and 13.3% after 1 day and 8, 20, and 32 months after incorporation of teliospores into soil, respectively, and was significantly (P < 0.01) affected by soil source. The percentage of teliospore recovery from soil was the greatest in loam soil and lowest from a silt loam soil. The rate of teliospores recovered from soil was not significantly affected by depth of burial and the soil source-depth interaction during the 32-month period. The percentage of germination of teliospores was significantly (P < 0.01) affected by soil source and depth of burial over the 32-month period. The mean percentage of teliospore germination at 1 day, and 8, 20, and 32 months after incorporation into soils was 51.3, 15.1, 16.4, and 16.5%, respectively. In another experiment, samples of silty clay loam soil with 5 × 103 teliospores of T. indica per gram of soil were stored at different temperatures in the laboratory. After 37 months of incubation at 22, 4, -5, and -18°C, the rates of teliospore recovered from soil were 1.6, 2.0, 5.7, and 11.3%, respectively. The percentage of spore germination from soil samples was highest at -5°C. Microscopy studies revealed that disintegration of teliospores begin after breakdown of the sheath-covering teliospore. The results of this study showed that teliospores of T. indica can survive in Montana for more than 32 months and remain viable.


The i5K Initiative: advancing arthropod genomics for knowledge, human health, agriculture, and the environment.
Insects and their arthropod relatives including mites, spiders, and crustaceans play major roles in the world's terrestrial, aquatic, and marine ecosystems. Arthropods compete with humans for food and transmit devastating diseases. They also comprise the most diverse and successful branch of metazoan evolution, with millions of extant species. Here, we describe an international effort to guide arthropod genomic efforts, from species prioritization to methodology and informatics. The 5000 arthropod genomes initiative (i5K) community met formally in 2012 to discuss a roadmap for sequencing and analyzing 5000 high-priority arthropods and is continuing this effort via pilot projects, the development of standard operating procedures, and training of students and career scientists. With university, governmental, and industry support, the i5K Consortium aspires to deliver sequences and analytical tools for each of the arthropod branches and each of the species having beneficial and negative effects on humankind.


Blazed grating fabrication through gray-scale Xray lithography.
Blazed gratings have been fabricated using gray-scale X-ray lithography. The gratings have high efficiency, low parasitic light, and high groove quality. The fabrication technique and resist characterization are described. The gratings can be generated over a considerable range of distances from the X-ray mask, thus demonstrating the ability to write gratings on a substrate of effectively arbitrary shape.


Developmental patterns of fetal fat and corresponding signal on T1-weighted magnetic resonance imaging.
Evaluation of subcutaneous fetal fat layer thickness on T1-weighted sequences can be used to predict birth weight. Little is known about normal MR signal patterns of subcutaneous tissue throughout pregnancy. To establish developmental patterns of subcutaneous fetal fat signal on T1-weighted sequences during the 2nd and 3rd trimesters. We retrospectively examined T1-weighted images of 110 fetal MRI scans. We measured signal intensity of subcutaneous fat on thighs, buttocks, trunk, nuchal region, chin and scalp. We then calculated the ratios of the obtained values with fetal muscle, amnios and maternal fat signal, and compared the results with those of immunohistochemical examination of adipose tissue extracted from the abdominal wall of fetuses as part of standard autopsy protocol. We included 60 MRI scans in fetuses without intra-uterine growth restriction or macrosomia of non-diabetic mothers (range 23-37 weeks of gestation). Fat T1 intensity of all anatomical regions was low in all fetuses before 26 weeks of gestation. It became more hyperintense with increasing gestational age, in the following order: chin and nuchal region, then buttocks, thighs and trunk, and eventually the scalp at 33 weeks of gestation. After 33 weeks of gestation, all fetal subcutaneous tissues demonstrated overall hyperintense signal. This progression followed the conversion at immunohistochemistry of fetal adipose tissue composition from predominant brown to white adipose cells in 19 fetuses (19-41 weeks of gestation). Between 26 weeks and 33 weeks of gestation, subcutaneous fetal fat signal changed in an orderly pattern from chin to buttocks and scalp. This may reflect the conversion from predominant brown to white adipose tissues in subcutaneous fetal fat.


The Effect of in vitro Aging and Fatigue on the Flexural Strength of Monolithic High-translucency Zirconia Restorations.
The aim of this in vitro study was to evaluate the effect of accelerated artificial aging (AAA) and fatigue on the biaxial flexural strength (BFS) of three types of monolithic high-translucency zirconia restorations compared with conventional low-translucency zirconia restorations. Four groups of 20 disc-shaped specimens (10 × 1.2 mm) were made from the following computer-aided design and computer-aided manufacturing (CAD/CAM) zirconia blocks: Low-translucency zirconia (Ceramill ZI-LT) as a control, and three brands of high-translucent zirconia (Lava Plus, Ceramill Zolid White, and Copran Monolithic HT). Ten discs from each group were subjected to the BFS test using the universal testing machine. The other 10 discs from each group were subjected to AAA (thermocycling, 3,500 cycles) and fatigue (250,000 cycles) before the fracture test. The definitive fracture load was recorded, and the BFS was calculated in accordance with International Organization for Standardization (ISO) 6872. The data were analyzed with one-way analysis of variance (ANOVA), Scheffe post hoc, and Mann-Whitney U test. Data analyses were evaluated at a significance level of p < 0.05. Significant differences were detected in the BFS among the four groups before AAA and fatigue. The mean BFS was highest with Ceramill ZI (935.3 ± 47.1 MPa), and least in Ceramill Zolid White (685.7 ± 32.6 MPa). After AAA and fatigue, significant differences were reported where the mean of BFS was highest with Copran Zr-i Monolithic HT (777.5 ± 21.2 MPa), and least in Ceramill Zolid White (576.0 ± 36.3 MPa). Furthermore, Mann-Whitney U test showed that AAA and fatigue significantly affect the BFS of each material individually. The AAA and fatigue significantly affected the BFS of the monolithic high-translucency zirconia restorations. Although monolithic high-translucency zirconia had significantly lower BFS than conventional zirconia tested in this study, they still have sufficient strength for clinical use.


The Rediscovery of Slowness: Exploring the Timing of Cognition.
Slowness of thought is not necessarily a handicap but could be a signature of optimal brain function. Emerging evidence shows that neuroanatomical and dynamical constraints of the human brain shape its functionality in optimal ways, characterized by slowness during task-based cognition in the context of spontaneous resting-state activity. This activity can be described mechanistically by whole-brain computational modeling that relates directly to optimality in the context of theories arguing for metastability in the brain. We discuss the role for optimal processing of information in the context of cognitive, task-related activity, and propose that combining multi-modal neuroimaging and explicit whole-brain models focused on the timing of functional dynamics can help to uncover fundamental rules of brain function in health and disease.


A psychometric assessment of the psychological and social well-being indicators in the PINE study.
This report aims to establish the reliability and validity of five psychological and social well-being indictors in their applications to a U.S. Chinese aging population. The Population Study of Chinese Elderly in Chicago (PINE) is a population-based epidemiological study of Chinese older adults in the Greater Chicago area. Internal consistency reliability was assessed by determining the coefficient alpha and inter-item correlation coefficients. Content validity was assessed by bilingual and bicultural study researchers and community leaders. The reliability and validity analysis supported the use of the Chinese version of the psychological and social well-being indicators in the PINE study. The value of Cronbach's alpha for fives scales ranged from .64 to .82, indicating reasonable internal consistency. Content validity was established through intensive review by a panel of experts. The instruments discussed in this report are reliable and valid measures to assess key dimensions of psychological and social distress of Chinese older adults.


Status of mitochondria in living human fibroblasts during growth and senescence in vitro: use of the laser dye rhodamine 123.
Rhodamine 123, a fluorescent laser dye that is selectively taken up into mitochondria of living cells, was used to examine mitochondrial morphology in early-passage (young), late-passage (old), and progeric human fibroblasts. Mitochondria were readily visualized in all cell types during growth (mid-log) and confluent stages. In all cell strains at confluence, mitochondria became shorter, more randomly aligned, and developed a higher proportion of bead-like forms. Treatment of cells for six days with Tevenel, a chloramphenicol analog that inhibits mitochondrial protein synthesis, brought about a marked depletion of mitochondria and a diffuse background fluorescence. Cyanide produced a rapid release of preloaded mitochondrial fluorescence followed by detachment and killing of cells. Colcemid caused a random coiling and fragmentation of mitochondria particularly in the confluent stage. No gross differences were discernible in mitochondria of the three cell strains in mid-log and confluent states or after these treatments. Butanol-extractable fluorescence after loading with rhodamine 123 was lower in all cell strains in confluent compared to mid-log stages. At confluence all three cell strains had similar rhodamine contents at zero-time and after washout up to 24 h. At the mid-log stage, young cells contained more rhodamine initially and lost it more rapidly than old or progeria cells, in that order. The data indicate no gross derangement in the morphology or number of mitochondria in old and progeria fibroblasts but there is a reduction of protonmotive force evident in these cells at the mid-log stage that may be growth limiting.


Recruited lysosomal enzymes as major digestive enzymes in insects.
The mass recruitment to the midgut contents of lysosomal proteolytic enzymes occurred in insects under three major selective pressures. Hemipteran (true bugs, aphids, and cicadas) ancestors lost their serine peptidases (SP) on adapting to feed on protein-free plant sap. When they returned to protein diets, their cathepsins L and B were recruited to replace their lost SP. Among beetles of the series Cucujiformia, cathepsins L were recruited to hydrolyze ingested plant inhibitors that affect their major SP and/or to deal with special seed proteins, such as prolamins. Larval flies have a very acid middle midgut and use cathepsin D to digest bacteria from their infected food. All the recruited enzymes originated from duplicated genes. The recruited digestive enzymes differ from their lysosomal counterparts in critical regions of their amino acid sequences that resulted in changes in substrate specificities and other kinetic properties. The discharge of digestive cathepsins in the midgut contents, instead of lysosomes, seems to be a consequence of their overexpression or the existence of new targeting signals. Their activation at the midgut contents occurs by an autoactivation mechanism or with the help of other enzymes or by a combination of both. The targeting to lysosomes of the insect lysosomal enzymes does not follow the mammalian mannose 6-phosphate route, but an incompletely known mechanism.


The effects of climate downscaling technique and observational data set on modeled ecological responses.
Assessments of future climate change impacts on ecosystems typically rely on multiple climate model projections, but often utilize only one downscaling approach trained on one set of observations. Here, we explore the extent to which modeled biogeochemical responses to changing climate are affected by the selection of the climate downscaling method and training observations used at the montane landscape of the Hubbard Brook Experimental Forest, New Hampshire, USA. We evaluated three downscaling methods: the delta method (or the change factor method), monthly quantile mapping (Bias Correction-Spatial Disaggregation, or BCSD), and daily quantile regression (Asynchronous Regional Regression Model, or ARRM). Additionally, we trained outputs from four atmosphere-ocean general circulation models (AOGCMs) (CCSM3, HadCM3, PCM, and GFDL-CM2.1) driven by higher (A1fi) and lower (B1) future emissions scenarios on two sets of observations (1/8º resolution grid vs. individual weather station) to generate the high-resolution climate input for the forest biogeochemical model PnET-BGC (eight ensembles of six runs).The choice of downscaling approach and spatial resolution of the observations used to train the downscaling model impacted modeled soil moisture and streamflow, which in turn affected forest growth, net N mineralization, net soil nitrification, and stream chemistry. All three downscaling methods were highly sensitive to the observations used, resulting in projections that were significantly different between station-based and grid-based observations. The choice of downscaling method also slightly affected the results, however not as much as the choice of observations. Using spatially smoothed gridded observations and/or methods that do not resolve sub-monthly shifts in the distribution of temperature and/or precipitation can produce biased results in model applications run at greater temporal and/or spatial resolutions. These results underscore the importance of carefully considering field observations used for training, as well as the downscaling method used to generate climate change projections, for smaller-scale modeling studies. Different sources of variability including selection of AOGCM, emissions scenario, downscaling technique, and data used for training downscaling models, result in a wide range of projected forest ecosystem responses to future climate change.


Determining the privacy policy deficiencies of health ICT applications through semi-formal modelling.
To ensure that patient confidentiality is securely maintained, health ICT applications that contain sensitive personal information demand comprehensive privacy policies. Determining the adequacy of these policies to meet legal conformity together with clinical users and patient expectation is demanding in practice. Organisations and agencies looking to analyse their Privacy and Security policies can benefit from guidance provided by outside entities such as the Privacy Office of their State or Government together with law firms and ICT specialists. The advice given is not uniform and often open to different interpretations. Of greater concern is the possibility of overlooking any important aspects that later result in a data breach. Based on three case studies, this paper considers whether a more formal approach to privacy analysis could be taken that would help identify the full coverage of a Privacy Impact Analysis and determine the deficiencies with an organisation's current policies and approach. A diagrammatic model showing the relationships between Confidentiality, Privacy, Trust, Security and Safety is introduced. First the validity of this model is determined by mapping it against the real-world case studies taken from three healthcare services that depend on ICT. Then, by using software engineering methods, a formal mapping of the relationships is undertaken to identify a full set of policies needed to satisfy the model. How effective this approach may prove as a generic method for deriving a comprehensive set of policies in health ICT applications is finally discussed.


Osteosarcoma diagnostic delay associated with alendronate-induced pain relief.
A 32-year-old man with a painful osteoblastic osteosarcoma of the right hip was initially diagnosed as having Paget's disease of bone. He was treated with alendronate for presumptive Paget's disease. The patient's bone pain was dramatically reduced by the administration of alendronate for 7 months. Following discontinuation of alendronate, his pain promptly recurred, culminating in a more thorough evaluation that led to the correct diagnosis. Despite chemotherapy, the patient succumbed to metastatic osteosarcoma. The main purpose of this publication is to report the potential for pain relief when an osteosarcoma is treated with bisphosphonate medication. Clinicians are advised not to consider an alendronate-associated pain reduction in an osteoblastic lesion as an indicator of an underlying benign process of bone. The evaluation of painful sclerotic bone lesions is briefly reviewed.


Time-dependent effects of pomegranate juice and pomegranate polyphenols on foodborne viral reduction.
Pomegranate juice (PJ) and pomegranate polyphenolic extracts (PP) have antiviral effects against HIV-1, influenza, herpes, and poxviruses, and we recently demonstrated their effect against human noroviral surrogates. In the present study, the time-dependent effects of two commercial brands of PJ and PP at two concentrations (2 and 4 mg/mL) on the infectivity of foodborne viral surrogates (feline calicivirus FCV-F9, murine norovirus MNV-1, and MS2 bacteriophage) at room temperature for up to 1 h were evaluated. Each virus at ∼5 log(10) plaque-forming units (PFU)/mL was mixed with equal volumes of PJ, or PP at 4 or 8 mg/mL, and incubated for 0, 10, 20, 30, 45, and 60 min at room temperature. Viral titers after each treatment were determined by standardized plaque assays and compared with untreated controls. Virus titer reduction by PJ and PP was found to be a rather rapid process, with ≥50% of titer reduction occurring within the first 20 min of treatment for all three tested viruses. Within the first 20 min, titer reductions of 3.12, 0.79, and 0.23 log(10) PFU/mL for FCV-F9, MNV-1, and MS2, respectively, were obtained using PJ. FCV-F9, MNV-1, and MS2 titers were reduced by 4.02, 0.68, and 0.18 log(10) PFU/mL with 2 mg/mL PP and 5.09, 1.14, and 0.19 log(10) PFU/mL with 4 mg/mL PP, respectively, after 20 min. The mechanism of viral reduction by PJ and PP needs to be elucidated and clinical trials should be undertaken before recommending for therapeutic or preventive purposes.


Abuse is not a zero-sum game! The case for zero tolerance of match official physical and verbal abuse.
Emergent research has investigated the impact of abuse on the decision of match officials to leave their sport. The existing literature is largely descriptive and qualitative. Based on large surveys of football referees in France and the Netherlands, this paper investigates the factors that are associated with the verbal and physical abuse of the referees and also the association of this abuse with the intentions of referees to quit officiating. The associations are investigated by estimating the marginal effects from bivariate probit and probit models respectively. Bivariate probit estimation reveals a strong correlation between each form of abuse. Both, unsurprisingly, are also positively associated with years of experience of referees. Probit estimation reveals that both forms of abuse, as well as intimidation from refereeing certain teams, are associated with an increased consideration of referees to quit. As increased intention to quit is also associated with the experience of the referee it is likely that the effect of abuse on referee considerations of quitting increase through time. The main conclusions are that the alternative forms of abuse are not zero-sum and both should be targeted by governing bodies to reduce the decline in the number of football referees. The data show that support of referees, for example through mentoring, can offset stated intentions to quit.HighlightsThis study looks at the factors that are associated with verbal and physical abuse of football referees and the association of this abuse with the intentions to quit.The alternative forms of abuse are not zero-sum and both should be targeted to reduce the decline in referees.Support of referees, for example through mentoring, can offset stated intentions to quit.


Toxicity associated with intensive postinduction therapy incorporating clofarabine in the very high-risk stratum of patients with newly diagnosed high-risk B-lymphoblastic leukemia: A report from the Children's Oncology Group study AALL1131.
Children, adolescents, and young adults with very high-risk (VHR) B acute lymphoblastic leukemia (B-ALL) have poor outcomes, and novel therapies are needed for this subgroup. The AALL1131 study evaluated postinduction therapy using cyclophosphamide (CPM), etoposide (ETOP), and clofarabine (CLOF) for patients with VHR B-ALL. Patients who were 1 to 30 years old and had VHR B-ALL received modified Berlin-Frankfurt-Münster therapy after induction and were randomized to 1) CPM, cytarabine, mercaptopurine, vincristine (VCR), and pegaspargase (control arm), 2) CPM, ETOP, VCR, and pegaspargase (experimental arm 1), or 3) CPM, ETOP, CLOF (30 mg/m2 /d × 5), VCR, and pegaspargase (experimental arm 2) during the second half of consolidation and delayed intensification. The rates of grade 4/5 infections and grade 3/4 pancreatitis were significantly increased in experimental arm 2. The dose of CLOF was, therefore, reduced to 20 mg/m2 /d × 5, and myeloid growth factor was required after CLOF administration. Despite these changes, 4 of 39 patients (10.3%) developed grade 4 infections, with 1 of these patients developing a grade 5 acute kidney injury attributed to CLOF, whereas only 1 of 46 patients (2.2%) in experimental arm 1 developed grade 4 infections, and there were no grade 4/5 infections in the control arm (n = 20). Four patients in experimental arm 2 had prolonged cytopenias for >60 days, whereas none did in the control arm or experimental arm 1. Counts failed to recover for 2 of these patients, one having a grade 5 acute kidney injury and the other removed from protocol therapy; both events occurred 92 days after the start of consolidation part 2. In AALL1131, CLOF, administered with CPM and ETOP, was associated with unacceptable toxicity. Cancer 2018;124:1150-9. © 2017 American Cancer Society.


Incentivising practice with take-home laparoscopic simulators in two UK Core Surgical Training programmes.
Practice using simulators has been validated as a mean for surgical trainees to improve basic laparoscopic skills and free their attention for higher cognitive functions. However, mere provision of equipment does not result in frequent practice. This study assesses one approach to incentivising practice within core surgical training programmes and leads to further recommendations. 30 core surgical trainees (CST) starting laparoscopic-based specialties were recruited from East and West of Scotland CST programmes and given take-home laparoscopic simulators, with six training modules. Attainment of target metric scores generated an eCertificate, to be rewarded by progression in the live theatre. Questionnaires assessed confounding variables and explored CSTs' anxieties about laparoscopy. 27 trainees (90%) agreed to participate (mean age 28 years, range 24-25; 17 males). 13 CSTs (48%) were in the first year of surgical training. 11 (41%) had no previous simulation experience and 7 (32%) CSTs played video games >3 hours/week. 12 of 27 trainees (44%) completed ≥1 task and 7 completed all (26%).Performances improved in some participants, but overall engagement with the programme was poor. Reasons given included poor internet connectivity, busy rotations and examinations. CSTs who engaged in the study significantly reduced their anxiety (mean 4.96 vs 3.56, p<0.05). The provision of take-home laparoscopic simulators with accompanying targets did not successfully incentivise CSTs to practise. However, the subgroup who did engage with the project reported performance improvements and significantly reduced anxiety. Proposals to overcome barriers to practising in simulation, including obligatory simulation-based assessments, are discussed.


L-carnitine prevents progression of non-alcoholic steatohepatitis in a mouse model with upregulation of mitochondrial pathway.
Non-alcoholic steatohepatitis (NASH) is a severe form of non-alcoholic fatty liver disease characterized by lobular inflammation, hepatocellular ballooning, and fibrosis with an inherent risk for progression to cirrhosis and hepatocellular carcinoma (HCC). Mitochondrial dysfunction appears to play a role in the progression from simple steatosis to NASH. L-carnitine (L-b-hydroxy-g-N-trimethylaminobutyric acid), an essential nutrient that converts fat into energy in mitochondria, has been shown to ameliorate liver damage. The aim of the present study was to explore the preventive and therapeutic effect of L-carnitine in NASH model mice. Eight-week-old male STAM mice, a NASH-cirrhosis-hepatocarcinogenic model, were divided into 3 experimental groups and fed as follows: 1) high-fat diet (HFD) (control group); 2) HFD mixed with 0.28% L-carnitine (L-carnitine group); and 3) HFD mixed with 0.01% α-tocopherol (α-tocopherol group). After 4 or 8 weeks, mice were sacrificed. Blood samples and livers were collected, and hepatic tumors were counted and measured. Livers were subjected to histological study, immunohistochemical staining of 4-hydroxynonenal and ferritin, determination of 8-OHdG levels, mRNA and protein expressions for multiple genes, and metabolomic analysis. The intestinal microbiome was also analyzed. L-carnitine increased hepatic expression of genes related to long-chain fatty acid transport, mitochondrial β-oxidation, and antioxidant enzymes following suppression of hepatic oxidative stress markers and inflammatory cytokines in NASH, and mice treated with L-carnitine developed fewer liver tumors. Although α-tocopherol resulted in NASH improvement in the same manner as L-carnitine, it increased periodontitis-related microbiotic changes and hepatic iron transport-related gene expression and led to less effective for anti-hepatocarcinogenesis. Conclusion: L-carnitine prevents progression of non-alcoholic steatohepatitis in a mouse model by upregulating the mitochondrial β-oxidation and redox system.


Effect of Prolonged Moderate Exercise on the Changes of Nonneuronal Cells in Early Myocardial Infarction.
Myocardial infarction (MI) is one of the leading causes of death in developed countries and it is characterized by several associated symptomatologies and poor quality of life. Recent data showed a possible interaction between infarction and brain inflammation and activity. Previous studies have demonstrated the beneficial effect of exercise training on deterioration in cardiac function after MI. In this study we analyzed in sedentary and trained rats the microglia and astrocytes 48 hours after MI in PVN, thalamus, prefrontal cortex, and hippocampus through immunofluorescence approach. We found significant changes in specific microglia phenotypes in the brain areas analyzed together with astrocytes activation. Prolonged exercise normalized these morphological changes of microglia and astrocytes in the prefrontal cortex, hippocampus, and thalamus but not in the PVN. Our data suggest that there is an early brain reaction to myocardial infarction induction, involving nonneuronal cells, that is attenuated by the prolonged exercise.


The effects of 5alpha-reductase inhibitors on the natural history, detection and grading of prostate cancer: current state of knowledge.
The Prostate Cancer Prevention Trial (PCPT) showed that the 5alpha-reductase inhibitor (5ARI) finasteride significantly decreased the 7-year period prevalence of prostate cancer vs placebo. However, Gleason score 7-10 tumors were significantly more common in the finasteride vs the placebo group. We considered data on the effects of 5ARIs on prostate cancer natural history and detection. A detailed review was performed of the literature identified from the MEDLINE database examining the effects of 5ARIs on prostate cancer prevalence and tumor histopathology. In PCPT there were fewer biopsies performed for cause in the finasteride vs the placebo group and the proportion of high grade tumors in the treatment groups did not diverge with time. Given that finasteride has an effect on prostate specific antigen and prostate volume, which are key factors in triggering prostate biopsies, they may be significant confounders of Gleason score results. Prostate shrinkage in the finasteride treated group may minimize biopsy sampling error. Furthermore, histological studies have shown that 5ARIs have a significant effect on prostate architecture, which can make the interpretation of prostate specimens in men treated with 5ARIs difficult. Further evaluation of PCPT findings will help determine the true nature of these observations. 5ARIs decrease the risk of prostate cancer but also alter the detection of disease through effects on prostate specific antigen, and prostate volume and histology. The weight of evidence suggests an artifactual effect of finasteride on Gleason grading in the PCPT. The role of 5ARIs for prostate cancer chemoprevention needs further examination before it can be considered for wide recommendation.


Fentanyl versus remifentanil-based TIVA for pediatric scoliosis repair: does it matter?
Opioid-induced hyperalgesia (OIH) and acute opioid tolerance have been demonstrated extensively in patients undergoing adolescent idiopathic scoliosis (AIS) repair. Remifentanil infusion has been strongly linked to both tolerance and OIH in these patients; however, the impact of using an intraoperative fentanyl infusion has not been well studied. This study aims to determine if patients undergoing operative management of AIS have decreased opioid consumption and pain scores when an intraoperative fentanyl infusion is used as compared with a remifentanil infusion. This is a retrospective chart review of patients with AIS who underwent posterior spinal fusion. During the period January 2012-June 2013, patients received remifentanil infusion as part of total intravenous anesthesia. From July 2013 to June 2015, remifentanil was replaced by fentanyl as standard protocol. The remifentanil cohort included 37 patients and the fentanyl cohort included 25 patients. The primary outcome was the total opioid consumption (morphine equivalents) in the first 24 hours postsurgery. Secondary outcomes included mean postoperative pain score in the first 24 hours postsurgery, postoperative opioid consumption 24-48 hours after surgery, time to extubation, time to assisted ambulation, length of stay, and incidence of postoperative nausea and vomiting. Compared with the remifentanil group, the fentanyl group had significantly higher postoperative opioid usage during the first 48 hours and significantly higher postoperative mean pain score during the first 24 hours. There was no difference between the two groups in mean pain score for 24-48 hours, extubation time, time to assisted ambulation, length of stay, or postoperative nausea and vomiting. Despite concerns for hyperalgesia and acute tolerance, remifentanil is widely used for intraoperative opioid infusions for surgical correction of AIS. This retrospective study examined a practice change from intraoperative remifentanil to intraoperative fentanyl as a potential approach to avoid OIH. Surprisingly, patients receiving fentanyl intraoperatively showed increased postoperative opioid use and pain scores in the first 24 hours postsurgery compared with the prior cohort receiving remifentanil. Substitution of fentanyl for remifentanil during surgical correction of AIS does not appear to solve the problem of OIH or acute tolerance. Prospective studies are needed to confirm this unexpected result.


Fatty acid transporter levels and palmitate oxidation rate correlate with ejection fraction in the infarcted rat heart.
Cardiac fatty acid uptake occurs predominantly via sarcolemmal transporter proteins; fatty acid translocase (FAT/CD36), plasma membrane fatty acid binding protein (FABPpm) and fatty acid transporter proteins (FATP) 1 and 6. We hypothesised that levels of the fatty acid transporters would be reduced in the chronically infarcted rat heart, in parallel with reduced dependence on fatty acid utilisation. In vivo left ventricular ejection fractions, measured using echocardiography, were 36% lower in rats six months after coronary artery ligation than in sham-operated control rats. In isolated, perfused, infarcted hearts, 3H-palmitate oxidation was 30% lower, and correlated with in vivo ejection fractions. As myocardial lipid incorporation was also reduced by 25%, total palmitate utilisation was 29% lower in the infarcted rat heart. The protein levels of the cardiac fatty acid transporters were reduced in the infarcted rat heart; FAT/CD36 by 36%, FABPpm by 12%, FATP6 by 21% and FATP1 by 26%, and the cytosolic fatty acid binding protein (cFABP) was 47% lower than in sham-operated rat hearts. Fatty acid transporter levels correlated with both palmitate oxidation rates and cardiac ejection fractions. Reductions in fatty acid oxidation and lipid incorporation rates were accompanied by downregulation of the cardiac fatty acid transporters. The metabolic shift away from fatty acid utilisation was proportional to the degree of functional impairment in the chronically infarcted rat heart.


Association of Abnormal Liver Function Parameters with HIV Serostatus and CD4 Count in Antiretroviral-Naive Rwandan Women.
We determined the associations of HIV infection/CD4 count with markers of hepatocellular damage [elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT)] and liver synthetic function (decreased albumin) in HIV-infected (HIV(+)) antiretroviral therapy (ART)-naive and uninfected (HIV(-)) Rwandan women. In 2005, 710 HIV(+) ART-naive and 226 HIV(-) women enrolled in the Rwanda Women's Interassociation Study and Assessment. Liver enzymes were measured with abnormality defined as either AST or ALT ≥1.25 times the upper limit of normal. Low serum albumin level was defined as <3.5 g/dl. Multivariable logistic regression analysis identified independent predictors of elevated AST/ALT and low serum albumin. HIV(-) women had the lowest prevalence (6.6%) of abnormal AST/ALT, with the highest prevalence (16.4%) in HIV(+) women with CD4 <200 cells/μl (p=0.01). The odds of having serum albumin <3.5 g/dl was 5.7-fold higher in HIV(+) than HIV(-) women (OR=5.68, 95% CI: 3.32-9.71). The risk of low albumin decreased from low to high CD4 count, with OR=2.62, 95% CI: 1.66, 4.14 and OR=1.57, 95% CI: 1.01, 2.43 in HIV(+) women with a CD4 count <200 and 200-350 cells/μl, respectively vs. HIV(+) with CD4 >350 (p<0.001 and p<0.05 for all comparisons). Our findings suggest that HIV-associated liver damage may occur in ART-naive patients. Although liver abnormality prevalences in this cohort of HIV-infected Rwandan women are less than reported in developed countries, caution is needed for risk assessment measures to monitor and screen HIV-infected patients pre- and post-ART initiation in African clinical settings to curtail potential risks associated with HIV infection.


Rapid growth of magnesium-carbonate weathering products in a stony meteorite from antarctica.
Nesquehonite, a hydrous magnesium carbonate, occurs as a weathering product on the surface of the Antarctic meteorite LEW 85320(H5 chondrite). Antarctic meteorites have resided on the earth for periods of 10(4) to 10(6) years, but the time needed for weathering products to form has been uncertain. Isotopic measurements of delta(13)C and delta(18)O indicate that the nesquehonite formed at near freezing temperatures by reaction of meteoritic minerals with terrestrial water and carbon dioxide. Results from carbon-14 dating suggest that, although the meteorite has been in Antarctica for at least 3.2 x 10(4) to 3.3 x 10(4) years, the nesquehonite formed after A.D.1950.


Quantifying Loss Caused by Ray Blight Disease in Tasmanian Pyrethrum Fields.
The efficacy of newly implemented fungicide recommendations on reducing the intensity of ray blight disease caused by Phoma ligulicola to achieve site-specific attainable yield potentials in Tasmanian pyrethrum fields was quantified over two seasons in 46 and 51 fields during the 2003 and 2004 growing seasons, respectively. Disease intensity and yield in two plots (10 × 24 m), one following the commercial fungicide protocol recommendations and the second receiving no fungicide, were assessed in each pyrethrum field. The commercial fungicide protocol consisted of one application of azoxystrobin at 150 g a.i./ha, followed by two applications of a tank mixture of difenoconazole at 125 g a.i./ha and chlorothalonil at 1,008 liters a.i./ha at 14- to 21-day intervals. This program resulted in significant decreases in defoliation severity and the incidence of stems and flowers with ray blight, and increases in the height of stems and number of flowers produced per stem in October and November. In plots receiving the commercial fungicide protocol, the dry weight of flowers was increased by 76 and 68% in 2003 and 2004, respectively. Moreover, pyrethrin yield increased by 81 and 78% when the commercial fungicide protocol was used compared with the nontreated plots. Tobit regression was used to examine the relationships and thresholds among disease intensity measures (defoliation severity, stem severity, and incidence of flowers with ray blight) assessed just prior to harvest. This regression utilized a left-censored regression model to define subminimal thresholds, as none of the disease intensity measures could be less than 0. Defoliation severity had a threshold of 35.3% before stem severity linearly increased and a threshold of 38.2% before the incidence of flowers with ray blight linearly increased. Finally, the threshold for stem severity was 13.7% before the incidence of flowers with ray blight linearly increased. These thresholds can be used to assist growers in making disease management decisions with the objective of minimizing loss of flowers by maintaining defoliation severity below the critical point at which the incidence of flowers with ray blight begins to linearly increase.


Charlemagne's summit canal: an early medieval hydro-engineering project for passing the Central European Watershed.
The Central European Watershed divides the Rhine-Main catchment and the Danube catchment. In the Early Medieval period, when ships were important means of transportation, Charlemagne decided to link both catchments by the construction of a canal connecting the Schwabian Rezat and the Altmühl rivers. The artificial waterway would provide a continuous inland navigation route from the North Sea to the Black Sea. The shortcut is known as Fossa Carolina and represents one of the most important Early Medieval engineering achievements in Europe. Despite the important geostrategic relevance of the construction it is not clarified whether the canal was actually used as a navigation waterway. We present new geophysical data and in situ findings from the trench fills that prove for the first time a total length of the constructed Carolingian canal of at least 2300 metres. We have evidence for a conceptual width of the artificial water course between 5 and 6 metres and a water depth of at least 60 to 80 cm. This allows a crossing way passage of Carolingian cargo scows with a payload of several tons. There is strong evidence for clayey to silty layers in the trench fills which reveal suspension load limited stillwater deposition and, therefore, the evidence of former Carolingian and post-Carolingian ponds. These findings are strongly supported by numerous sapropel layers within the trench fills. Our results presented in this study indicate an extraordinarily advanced construction level of the known course of the canal. Here, the excavated levels of Carolingian trench bottoms were generally sufficient for the efficient construction of stepped ponds and prove a final concept for a summit canal. We have evidence for the artificial Carolingian dislocation of the watershed and assume a sophisticated Early Medieval hydrological engineering concept for supplying the summit of the canal with adequate water.


From segment to somite: segmentation to epithelialization analyzed within quantitative frameworks.
One of the most visually striking patterns in the early developing embryo is somite segmentation. Somites form as repeated, periodic structures in pairs along nearly the entire caudal vertebrate axis. The morphological process involves short- and long-range signals that drive cell rearrangements and cell shaping to create discrete, epithelialized segments. Key to developing novel strategies to prevent somite birth defects that involve axial bone and skeletal muscle development is understanding how the molecular choreography is coordinated across multiple spatial scales and in a repeating temporal manner. Mathematical models have emerged as useful tools to integrate spatiotemporal data and simulate model mechanisms to provide unique insights into somite pattern formation. In this short review, we present two quantitative frameworks that address the morphogenesis from segment to somite and discuss recent data of segmentation and epithelialization.


Is waist circumference per body mass index rising differentially across the United States, England, China and Mexico?
Little is known about whether waist circumference (WC) has increased disproportionately relative to body mass index (BMI) around the world. Data came from the US National Health and Nutrition Examination Survey (1988-1994 and 2007-2010), Health Survey for England (1992-1993 and 2008-2009); the Mexican Nutrition Survey (1999) and the Mexican National Health and Nutrition Survey (NHNS 2012); and the China Health and Nutrition Survey (1993 and 2011). Country- and sex-stratified (for the United States, also race-/ethnicity-stratified) multivariable linear regressions were used to estimate mean difference in WC over time relative to BMI at specified overweight and obesity cutoff points, adjusting for age and survey year. Although mean WC and BMI shifted upward over time in all age-sex subpopulations in all four countries, trends in overweight prevalence were less consistent. However, WC relative to BMI increased at varying magnitudes across all countries and subpopulations, except US Black men. The magnitude of increase was largest for women in the youngest age group (20-29 years), particularly for women in Mexico (+6.6 cm, P<0.0001) and China (+4.6 cm, P<0.0001) (holding BMI constant at 25 kg/m(2)). For men, the increase was primarily evident among Chinese men (+4.8 cm, P<0.0001). WC has increased disproportionately over time relative to overall body mass across the United States, England, Mexico and China, particularly among young women, with the largest increases occurring in the middle-income countries of Mexico and China. These patterns are potentially a cause for concern especially for countries undergoing rapid economic and nutritional transitions.


CRISPR/Cas9-mediated homology-directed repair by ssODNs in zebrafish induces complex mutational patterns resulting from genomic integration of repair-template fragments.
Targeted genome editing by CRISPR/Cas9 is extremely well fitted to generate gene disruptions, although precise sequence replacement by CRISPR/Cas9-mediated homology-directed repair (HDR) suffers from low efficiency, impeding its use for high-throughput knock-in disease modeling. In this study, we used next-generation sequencing (NGS) analysis to determine the efficiency and reliability of CRISPR/Cas9-mediated HDR using several types of single-stranded oligodeoxynucleotide (ssODN) repair templates for the introduction of disease-relevant point mutations in the zebrafish genome. Our results suggest that HDR rates are strongly determined by repair-template composition, with the most influential factor being homology-arm length. However, we found that repair using ssODNs does not only lead to precise sequence replacement but also induces integration of repair-template fragments at the Cas9 cut site. We observed that error-free repair occurs at a relatively constant rate of 1-4% when using different repair templates, which was sufficient for transmission of point mutations to the F1 generation. On the other hand, erroneous repair mainly accounts for the variability in repair rate between the different repair templates. To further improve error-free HDR rates, elucidating the mechanism behind this erroneous repair is essential. We show that the error-prone nature of ssODN-mediated repair, believed to act via synthesis-dependent strand annealing (SDSA), is most likely due to DNA synthesis errors. In conclusion, caution is warranted when using ssODNs for the generation of knock-in models or for therapeutic applications. We recommend the application of in-depth NGS analysis to examine both the efficiency and error-free nature of HDR events.This article has an associated First Person interview with the first author of the paper.


Influence of phospholipid composition on self-assembly and energy-transfer efficiency in networks of light-harvesting 2 complexes.
In the photosynthetic membrane of purple bacteria networks of light-harvesting 2 (LH2) complexes capture the sunlight and transfer the excitation energy. In order to investigate the mutual relationship between the supramolecular organization of the pigment-protein complexes and their biological function, the LH2 complexes were reconstituted into three types of phospholipid membranes, consisting of L-α-phosphatidylglycerol (PG), L-α-phosphatidylcholine (PC), and L-α-phosphatidylethanolamine (PE)/PG/cardiolipin (CL). Atomic force microscopy (AFM) revealed that the type of phospholipids had a crucial influence on the clustering tendency of the LH2 complexes increased from PG over PC to PE/PG/CL, where the LH2 complexes formed large, densely packed clusters. Time-resolved spectroscopy uncovered a strong quenching of the LH2 fluorescence that is ascribed to singlet-singlet and singlet-triplet annihilation by an efficient energy transfer between the LH2 complexes in the artificial membrane systems. Quantitative analysis reveals that the intercomplex energy transfer efficiency varies strongly as a function of the morphology of the nanostructure, namely in the order PE/PG/CL > PC > PG, which is in line with the clustering tendency of LH2 observed by AFM. These results suggest a strong influence of the phospholipids on the self-assembly of LH2 complexes into networks and concomitantly on the intercomplex energy transfer efficiency.


The Spontaneous Mutation Rate in the Fission Yeast Schizosaccharomyces pombe.
The rate at which new mutations arise in the genome is a key factor in the evolution and adaptation of species. Here we describe the rate and spectrum of spontaneous mutations for the fission yeast Schizosaccharomyces pombe, a key model organism with many similarities to higher eukaryotes. We undertook an ∼1700-generation mutation accumulation (MA) experiment with a haploid S. pombe, generating 422 single-base substitutions and 119 insertion-deletion mutations (indels) across the 96 replicates. This equates to a base-substitution mutation rate of 2.00 × 10(-10) mutations per site per generation, similar to that reported for the distantly related budding yeast Saccharomyces cerevisiae. However, these two yeast species differ dramatically in their spectrum of base substitutions, the types of indels (S. pombe is more prone to insertions), and the pattern of selection required to counteract a strong AT-biased mutation rate. Overall, our results indicate that GC-biased gene conversion does not play a major role in shaping the nucleotide composition of the S. pombe genome and suggest that the mechanisms of DNA maintenance may have diverged significantly between fission and budding yeasts. Unexpectedly, CpG sites appear to be excessively liable to mutation in both species despite the likely absence of DNA methylation.


Computer-based learning of spelling skills in children with and without dyslexia.
Our spelling training software recodes words into multisensory representations comprising visual and auditory codes. These codes represent information about letters and syllables of a word. An enhanced version, developed for this study, contains an additional phonological code and an improved word selection controller relying on a phoneme-based student model. We investigated the spelling behavior of children by means of learning curves based on log-file data of the previous and the enhanced software version. First, we compared the learning progress of children with dyslexia working either with the previous software (n = 28) or the adapted version (n = 37). Second, we investigated the spelling behavior of children with dyslexia (n = 37) and matched children without dyslexia (n = 25). To gain deeper insight into which factors are relevant for acquiring spelling skills, we analyzed the influence of cognitive abilities, such as attention functions and verbal memory skills, on the learning behavior. All investigations of the learning process are based on learning curve analyses of the collected log-file data. The results evidenced that those children with dyslexia benefit significantly from the additional phonological cue and the corresponding phoneme-based student model. Actually, children with dyslexia improve their spelling skills to the same extent as children without dyslexia and were able to memorize phoneme to grapheme correspondence when given the correct support and adequate training. In addition, children with low attention functions benefit from the structured learning environment. Generally, our data showed that memory sources are supportive cognitive functions for acquiring spelling skills and for using the information cues of a multi-modal learning environment.


Molecular basis for solvent dependent morphologies observed on electrosprayed surfaces.
We study the causes of the observed tunable hydrophobicity of poly(styrene-co-perfluoroalkyl ethylacrylate) electrosprayed in THF, DMF, and THF:DMF (1:1) solvents. Under the assumption that equilibrium morphologies in the solvent significantly affect the patterns observed on electrosprayed surfaces, we use atomistic and coarse-grained simulations supported by dynamic light scattering (DLS) experiments to focus on the parameters that affect the resulting morphology of superhydrophobic electrosprayed beads. The differing equilibrium chain size distributions in these solvents examined by DLS are corroborated by chain dimensions obtained via molecular dynamics simulations. Mesoscopic morphologies monitored by dissipative particle dynamics simulations explain experimental observations; in particular, the preference of the polymer for THF over DMF in the binary mixture rationalizes the dual scale roughness driven by stable microphase separation. Drying phenomena that affect resultant dual-scale roughness are described in three stages, each interpreted by concentration dependent diffusion and surface mass transfer coefficients of the solvents. Irrespective of the presence of polar groups in the structure, a conflict between the lower-boiling point solvent adhering to the polymer and the less volatile solvent abundant in the bulk leads to perfectly hydrophobic surfaces.


Surgical management of BCG vaccine-induced regional axillary lymphadenitis in HIV-infected children.
There are as yet no clear surgical guidelines for the management of BCG vaccine-induced regional axillary lymphadenopathy. The aim of this study was to evaluate the management of the condition and to suggest possible management strategies. A retrospective study was undertaken of 23 cases of suspected ipsilateral BCG adenitis following neonatal BCG inoculation (2001 - 2004). Diagnosis of a BCG infection was confirmed by culture and/or gastric washout. The age of the patient and mode of presentation, imaging findings, and results of tuberculin skin testing (Mantoux test) were documented. Because of a change in management policy the first group of patients treated by primary surgery were compared with those treated by fine-needle aspiration (FNA). The influence of HIV status on outcome was assessed. Surgical complications and outcome were analysed. Twenty-three children under 13 years of age (mean age 8.8 months, male/female ratio 1.9:1) were evaluated. Eighteen patients tested positive for HIV and 5 were HIV-negative. A positive culture for BCG bacillus was identified in 19 cases (83%) - by FNA (N=13, 68%), on pus swab (N=3, 16%), at surgery (N=1, 5%), and by gastric washing (N=2, 11%). Three HIV-negative children had granulomas on histological examination without a positive culture. Forty-five per cent of the 11 patients treated early in the study period by primary surgery (drainage/biopsy) had complications, which included a difficult anaesthetic induction and technical surgical difficulties. The postoperative incidence of wound dehiscence/infection was extremely high in this group and 18.2% developed postoperative cutaneous sinuses. Following a change in management policy, the following 12 patients, with a comparable HIV incidence, treated by initial conservative management, had a much lower incidence of post-procedural complications. This study confirms a high perioperative complication rate associated with the primary surgical treatment of BCG lymphadenitis in both HIV-positive and negative patients. Primary surgical treatment (incisional drainage or biopsy) is therefore not considered an ideal form of management in BCG lymphadenitis because of the high fistulisation and poor wound healing, especially in the HIV-positive patient. It should be avoided as the initial approach, with needle aspiration being preferred. Surgery should therefore be confined to the unusual event of real doubt about the underlying diagnosis and the treatment of suppurative complications.


Fracture prediction, imaging and screening in osteoporosis.
Osteoporosis is associated with increased fragility of bone and a subsequent increased risk of fracture. The diagnosis of osteoporosis is intimately linked with the imaging and quantification of bone and BMD. Scanning modalities, such as dual-energy X-ray absorptiometry or quantitative CT, have been developed and honed over the past half century to provide measures of BMD and bone microarchitecture for the purposes of clinical practice and research. Combined with fracture prediction tools such as Fracture Risk Assessment Tool (FRAX) (which use a combination of clinical risk factors for fracture to provide a measure of risk), these elements have led to a paradigm shift in the ability to diagnose osteoporosis and predict individuals who are at risk of fragility fracture. Despite these developments, a treatment gap exists between individuals who are at risk of osteoporotic fracture and those who are receiving therapy. In this Review, we summarize the epidemiology of osteoporosis, the history of scanning modalities, fracture prediction tools and future directions, including the most recent developments in prediction of fractures.


Suicide and schizophrenia: a systematic review of rates and risk factors.
Risk assessment is a core skill in psychiatry. Risk prediction for suicide in schizophrenia is known to be complex. We undertook a systematic review of all original studies concerning suicide in schizophrenia published since 2004. We found 51 data-containing studies (from 1281 studies screened) that met our inclusion criteria, and ranked these by standardized quality criteria. Estimates of rates of suicide and risk factors associated with later suicide were identified, and the risk factors were grouped according to type and strength of association with suicide. Consensus on the lifetime risk of suicide was a rate of approximately 5%. Risk factors with a strong association with later suicide included being young, male, and with a high level of education. Illness-related risk factors were important predictors, with number of prior suicide attempts, depressive symptoms, active hallucinations and delusions, and the presence of insight all having a strong evidential basis. A family history of suicide, and comorbid substance misuse were also positively associated with later suicide. The only consistent protective factor for suicide was delivery of and adherence to effective treatment. Prevention of suicide in schizophrenia will rely on identifying those individuals at risk, and treating comorbid depression and substance misuse, as well as providing best available treatment for psychotic symptoms.


Mindfulness-based cognitive therapy may reduce thought suppression in previously suicidal participants: findings from a preliminary study.
Thought suppression is a strategy aimed at mental control that may paradoxically increase the frequency of unwanted thoughts. This preliminary study examined effects of mindfulness-based cognitive therapy (MBCT) on thought suppression and depression in individuals with past depression and suicidality. In a randomized controlled trial design, 68 participants were allocated to an MBCT group or a treatment-as-usual waitlist control. Measures of thought suppression and depression were taken pre- and post-treatment. MBCT did not reduce thought suppression as measured by the White Bear Suppression Inventory, but significantly reduced self-reported attempts to suppress in the previous week. Preliminary evidence suggests that MBCT for suicidality may reduce thought suppression, but differential effects on thought suppression measures warrant further studies.


Stochastic memristive devices for computing and neuromorphic applications.
Nanoscale resistive switching devices (memristive devices or memristors) have been studied for a number of applications ranging from non-volatile memory, logic to neuromorphic systems. However a major challenge is to address the potentially large variations in space and time in these nanoscale devices. Here we show that in metal-filament based memristive devices the switching can be fully stochastic. While individual switching events are random, the distribution and probability of switching can be well predicted and controlled. Rather than trying to force high switching probabilities using excess voltage or time, the inherent stochastic nature of resistive switching allows these binary devices to be used as building blocks for novel error-tolerant computing schemes such as stochastic computing and provides the needed "analog" feature for neuromorphic applications. To verify such potential, we demonstrated memristor-based stochastic bitstreams in both time and space domains, and show that an array of binary memristors can act as a multi-level "analog" device for neuromorphic applications.


Effects of a cyclo-oxygenase inhibitor, flurbiprofen, and an H1 histamine receptor antagonist, terfenadine, alone and in combination on allergen induced immediate bronchoconstriction in man.
The effect of flurbiprofen, a potent cyclo-oxygenase inhibitor, on histamine and methacholine reactivity was assessed in seven atopic subjects with asthma. Flurbiprofen 150 mg daily for three days displaced the histamine-FEV1 concentration-response curve to the right by 1.5 doubling doses, whereas no effect was observed on the response to methacholine. Subsequently the effects of flurbiprofen and terfenadine, a specific H1 histamine receptor antagonist, on allergen induced bronchoconstriction were studied in seven atopic but non-asthmatic subjects. The subjects inhaled the concentration of grass pollen allergen that had previously been shown to produce a 20% fall in FEV1 on separate occasions after prior treatment with placebo, flurbiprofen 150 mg daily for three days, terfenadine 180 mg three hours before challenge, and the combination of flurbiprofen and terfenadine. After placebo, allergen challenge caused a mean (SEM) maximum fall in FEV1 of 37.6% (2.6%) after 20 (3.7) minutes, followed by a gradual recovery to within 15% of baseline at 60 minutes. Terfenadine reduced the maximum allergen provoked fall in FEV1 to 21.5% (2.2%) and reduced the area under the time-response curve (AUC) by 50% (6%). Flurbiprofen alone reduced the mean maximum fall in FEV1 to 29.6% (3.2%) and reduced the AUC by 26%. The effect of the combination of flurbiprofen and terfenadine did not differ significantly from that of terfenadine alone. We conclude that histamine and prostaglandins contribute to immediate allergen induced bronchoconstriction and that a complex interaction occurs between the two classes of mediators.


Plasma metabolomics reveals a diagnostic metabolic fingerprint for mitochondrial aconitase (ACO2) deficiency.
Mitochondrial respiratory chain dysfunction has been identified in a number of neurodegenerative disorders. Infantile cerebellar-retinal degeneration associated with mutations in the mitochondrial aconitase 2 gene (ACO2) has been recently described as a neurodegenerative disease of autosomal recessive inheritance. To date there is no biomarker for ACO2 deficiency and diagnosis relies on genetic analysis. Here we report global metabolic profiling in eight patients with ACO2 deficiency. Using an LC-MS-based metabolomics platform we have identified several metabolites with affected plasma concentrations including the tricarboxylic acid cycle metabolites cis-aconitate, isocitrate and alpha-ketoglutarate, as well as phosphoenolpyruvate and hydroxybutyrate. Taken together we report a diagnostic metabolic fingerprint for mitochondrial aconitase 2 deficiency.


Functional metagenomic analysis reveals rivers are a reservoir for diverse antibiotic resistance genes.
The environment harbours a significant diversity of uncultured bacteria and a potential source of novel and extant resistance genes which may recombine with clinically important bacteria disseminated into environmental reservoirs. There is evidence that pollution can select for resistance due to the aggregation of adaptive genes on mobile elements. The aim of this study was to establish the impact of waste water treatment plant (WWTP) effluent disposal to a river by using culture independent methods to study diversity of resistance genes downstream of the WWTP in comparison to upstream. Metagenomic libraries were constructed in Escherichia coli and screened for phenotypic resistance to amikacin, gentamicin, neomycin, ampicillin and ciprofloxacin. Resistance genes were identified by using transposon mutagenesis. A significant increase downstream of the WWTP was observed in the number of phenotypic resistant clones recovered in metagenomic libraries. Common β-lactamases such as blaTEM were recovered as well as a diverse range of acetyltransferases and unusual transporter genes, with evidence for newly emerging resistance mechanisms. The similarities of the predicted proteins to known sequences suggested origins of genes from a very diverse range of bacteria. The study suggests that waste water disposal increases the reservoir of resistance mechanisms in the environment either by addition of resistance genes or by input of agents selective for resistant phenotypes.


Reactivity of selenium-containing compounds with myeloperoxidase-derived chlorinating oxidants: Second-order rate constants and implications for biological damage.
Hypochlorous acid (HOCl) and N-chloramines are produced by myeloperoxidase (MPO) as part of the immune response to destroy invading pathogens. However, MPO also plays a detrimental role in inflammatory pathologies, including atherosclerosis, as inappropriate production of oxidants, including HOCl and N-chloramines, causes damage to host tissue. Low molecular mass thiol compounds, including glutathione (GSH) and methionine (Met), have demonstrated efficacy in scavenging MPO-derived oxidants, which prevents oxidative damage in vitro and ex vivo. Selenium species typically have greater reactivity toward oxidants compared to the analogous sulfur compounds, and are known to be efficient scavengers of HOCl and other hypohalous acids produced by MPO. In this study, we examined the efficacy of a number of sulfur and selenium compounds to scavenge a range of biologically relevant N-chloramines and oxidants produced by both isolated MPO and activated neutrophils and characterized the resulting selenium-derived oxidation products in each case. A dose-dependent decrease in the concentration of each N-chloramine was observed on addition of the sulfur compounds (cysteine, methionine) and selenium compounds (selenomethionine, methylselenocysteine, 1,4-anhydro-4-seleno-L-talitol, 1,5-anhydro-5-selenogulitol) studied. In general, selenomethionine was the most reactive with N-chloramines (k2 0.8-3.4×10(3)M(-1) s(-1)) with 1,5-anhydro-5-selenogulitol and 1,4-anhydro-4-seleno-L-talitol (k2 1.1-6.8×10(2)M(-1) s(-1)) showing lower reactivity. This resulted in the formation of the respective selenoxides as the primary oxidation products. The selenium compounds demonstrated greater ability to remove protein N-chloramines compared to the analogous sulfur compounds. These reactions may have implications for preventing cellular damage in vivo, particularly under chronic inflammatory conditions.


Rapid 20th-century increase in coastal upwelling off northwest Africa.
Near-shore waters along the northwest African margin are characterized by coastal upwelling and represent one of the world's major upwelling regions. Sea surface temperature (SST) records from Moroccan sediment cores, extending back 2500 years, reveal anomalous and unprecedented cooling during the 20th century, which is consistent with increased upwelling. Upwelling-driven SSTs also vary out of phase with millennial-scale changes in Northern Hemisphere temperature anomalies (NHTAs) and show relatively warm conditions during the Little Ice Age and relatively cool conditions during the Medieval Warm Period. Together, these results suggest that coastal upwelling varies with NHTAs and that upwelling off northwest Africa may continue to intensify as global warming and atmospheric CO2 levels increase.


The innovative regularity and role of p16 methylation in blood during HCC development.
Purpose: This study was performed to examine the regularity and role of p16 methylation in hepatocellular carcinoma (HCC) blood. Methods: Big data of the case-control studies due to blood p16 methylation detection were collected in English and Chinese Journals. The risk of HCC's histologic process was investigated using both Meta-analysis and the quantitative correlation analysis. Results: p16 methylation frequencies in blood were gradually increased from 0 % in normal to 10 % in benign disease, and to 60 % in HCC development. Based on p16 methylation of normal control blood, p16 methylation between normal and benign disease had no risk, and the methylation risk in HCC was significantly increased from normal to HCC through benign disease OR, 95% CI =16.23 ( 11.66, 22.58 ). Compared with the benign disease matched by HCC patient, the methylation risk of p16 in HCC was found, with the pooled OR value of 10.06 (95% IC = 7.64, 13.21) in blood. In addition, the regulatory mechanism affecting p16 methylation risk in normal blood had no role, and the strength of p16 methylation risk was rapidly increased between benign diseases and HCC blood. p16 methylation risk started from the patients with benign disease in blood. These results in blood and tissue detection were basically consistent. Conclusions: HCC pathogenesis affecting p16 methylation don't work during normal blood, when from benign diseases to HCC bloods, can produce powerful role. The transcriptional inactivation associated with p16 methylation might start from benign liver disease, and might be increased from benign liver disease to HCC process. p16 methylation in blood can be used as a promising non-invasive biomarker to HCC's prediction and diagnosis.


Antibody to a 39-kilodalton Borrelia burgdorferi antigen (P39) as a marker for infection in experimentally and naturally inoculated animals.
Borrelia burgdorferi expresses a conserved, species-specific 39-kDa protein (P39) that can stimulate antibodies during human infection. To confirm that anti-P39 antibodies are produced consistently in animals exposed to infectious spirochetes, white-footed mice, Peromyscus leucopus, and laboratory white mice, Mus musculus (strain BALB/c), were experimentally inoculated with either infectious or noninfectious B. burgdorferi and the antibody response to P39 was determined by immunoblot at 21 days postinoculation. All mice inoculated with approximately 10(7) infectious B. burgdorferi produced anti-P39 antibodies and were cultured positive for this spirochete. Mice inoculated with similar numbers of inactivated or viable noninfectious B. burgdorferi still producing P39 did not induce anti-P39 antibodies. By contrast, putative antiflagellin antibodies were detected in less than 18% of the infected animals, which supports the notion that antibody reactive with flagellin may not be reliable as a marker for B. burgdorferi exposure as was originally thought. Mice infected with B. burgdorferi following exposure to ticks (Ixodes dammini) produced anti-P39 antibodies no later than 7 days postinfection, indicating that P39 is an effective immunogen in natural infections. Notably, anti-P39 antibodies were the predominant B. burgdorferi reactive antibodies detected early in the infection. Our results indicate that anti-P39 antibodies are produced in response to an active infection and are therefore reliable markers for infection in experimentally and naturally inoculated animals.


Population structure of Rudgea parquioides, a shade-tolerant shrub species, in Southern Brazil.
The expectations that shade-tolerant forest species show 1) a population structure composed by a high amount of small individuals, and 2) biomass allocation for diameter higher than for height growth, were tested for Rudgea parquioides, a typical shrub in Southern Brazil. We described the size structure (height and stem diameter) and allometrical relations of a R. parquioides population by counting and measuring all the individuals in a 725m(2) area in the municipality of Curitiba (25°25'S; 49°19'W). A total of 916 individuals (12,634 ind.ha(-1)) were recorded in the area. The first expectation was supported, since distribution by height and diameter classes showed a predominance of small individuals (skewness coefficients > 1). On the other hand, the regression between height and stem base diameter showed slope β < 1, which indicates that growth in height is higher than in diameter, not supporting the second expectation. These results show that life strategies in shade-tolerant species may imply in more trade-off combinations than previously described.


The players in a mutualistic symbiosis: insects, bacteria, viruses, and virulence genes.
Aphids maintain mutualistic symbioses involving consortia of coinherited organisms. All possess a primary endosymbiont, Buchnera, which compensates for dietary deficiencies; many also contain secondary symbionts, such as Hamiltonella defensa, which confers defense against natural enemies. Genome sequences of uncultivable secondary symbionts have been refractory to analysis due to the difficulties of isolating adequate DNA samples. By amplifying DNA from hemolymph of infected pea aphids, we obtained a set of genomic sequences of H. defensa and an associated bacteriophage. H. defensa harbors two type III secretion systems, related to those that mediate host cell entry by enteric pathogens. The phage, called APSE-2, is a close relative of the previously sequenced APSE-1 but contains intact homologs of the gene encoding cytolethal distending toxin (cdtB), which interrupts the eukaryotic cell cycle and which is known from a variety of mammalian pathogens. The cdtB homolog is highly expressed, and its genomic position corresponds to that of a homolog of stx (encoding Shiga-toxin) within APSE-1. APSE-2 genomes were consistently abundant in infected pea aphids, and related phages were found in all tested isolates of H. defensa, from numerous insect species. Based on their ubiquity and abundance, these phages appear to be an obligate component of the H. defensa life cycle. We propose that, in these mutualistic symbionts, phage-borne toxin genes provide defense to the aphid host and are a basis for the observed protection against eukaryotic parasites.


Neuroprotective mesenchymal stem cells are endowed with a potent antioxidant effect in vivo.
Experimental autoimmune encephalomyelitis (EAE), an animal model for human multiple sclerosis, is characterized by demyelination, inflammation and neurodegeneration of CNS in which free radicals play a role. Recently, the efficacy of murine mesenchimal stem cells (MSCs) as treatment of EAE induced in mice by the encephalitogenic peptide MOG(35-55) was demonstrated. The present study analyzed some markers of oxidative stress, inflammation/degeneration and apoptosis such as metallothioneins (MTs), antioxidant enzymes (superoxide dismutase, catalase and glutathione-S-transferase), poly(ADP-ribose) polymerase-1 and p53 during EAE progression and following MSC treatment. Expression of the three brain MT isoforms increased significantly in EAE mice compared with healthy controls, but while expression of MT-1 and MT-3 increased along EAE course, MT-2 was up-regulated at the onset, but returned to levels similar to those of controls in chronic phase. The changes in the transcription and activity of the antioxidant enzymes and in expression of poly(ADP-ribose) polymerase-1 and p53 showed the same kinetics observed for MT-1 and MT-3 during EAE. Interestingly, i.v. administration of MSCs reduced the EAE-induced increases in levels/activities of all these proteins. These results support an antioxidant and neuroprotective activity for MSCs that was also confirmed in vitro on neuroblastoma cells exposed to an oxidative insult.


The role of the rehabilitation brace in restoring knee extension after anterior cruciate ligament reconstruction: a prospective controlled study.
To evaluate the effects of a rehabilitation brace locked in extension for the first week on the recovery of full extension after ACL reconstruction we compared two groups of subjects who underwent ACL bone-patellar tendon-bone reconstruction. The brace was unlocked twice a day for assisted physiotherapy. In Group A, 18 male subjects wore a post-operative brace locked from 0 degrees to 90 degrees. In Group B, 18 male subjects wore the same post-op brace locked in full extension for the first week. In both groups the brace was unlocked (0 degrees -120 degrees ) at the beginning of the second postoperative week, and then removed at the beginning of the third week. All the subjects followed the same rigorous, accelerated rehabilitation protocol. Each subject was evaluated pre and post-operatively (at second, fourth and eighth week) with bubble-level heel height difference (HHD) measurements and KT 1000 arthrometric assessment at the fourth postoperative month. Preoperative bubble-level HHD measurements of the two groups were statistically similar (Group A 0.6 cm, Group B 1 cm; not significant, n.s.). At the fourth week (Group A 2.2 cm, Group B 0.6 cm) and eighth week (Group A 1.6 cm, Group B 0.1 cm) follow-ups, bubble-level HHD measurements showed that the extension of the operated knees of Group B was significantly greater than in Group A. KT 1000 arthrometric scores showed no difference between the two groups (Group A 1.8 mm, Group B 1.5; n.s.).


Silicon Nanofluidic Membrane for Electrostatic Control of Drugs and Analytes Elution.
Individualized long-term management of chronic pathologies remains an elusive goal despite recent progress in drug formulation and implantable devices. The lack of advanced systems for therapeutic administration that can be controlled and tailored based on patient needs precludes optimal management of pathologies, such as diabetes, hypertension, rheumatoid arthritis. Several triggered systems for drug delivery have been demonstrated. However, they mostly rely on continuous external stimuli, which hinder their application for long-term treatments. In this work, we investigated a silicon nanofluidic technology that incorporates a gate electrode and examined its ability to achieve reproducible control of drug release. Silicon carbide (SiC) was used to coat the membrane surface, including nanochannels, ensuring biocompatibility and chemical inertness for long-term stability for in vivo deployment. With the application of a small voltage (≤ 3 V DC) to the buried polysilicon electrode, we showed in vitro repeatable modulation of membrane permeability of two model analytes-methotrexate and quantum dots. Methotrexate is a first-line therapeutic approach for rheumatoid arthritis; quantum dots represent multi-functional nanoparticles with broad applicability from bio-labeling to targeted drug delivery. Importantly, SiC coating demonstrated optimal properties as a gate dielectric, which rendered our membrane relevant for multiple applications beyond drug delivery, such as lab on a chip and micro total analysis systems (µTAS).


Resolving the taxonomic conundrum in Graphoderus of the east Palearctic with a key to all species (Coleoptera, Dytiscidae).
The Holarctic diving beetle genus Graphoderus (Dytiscinae, Aciliini) contains relatively few and well-known species but these may still be difficult to identify based on external characters. A taxonomic problem in the eastern Palearctic was discovered that relates to the Palearctic Graphoderus zonatus (Hoppe, 1795) and the Nearctic Graphoderus perplexus Sharp, 1882. Based on qualitative and quantitative characters, especially on male genitalia which have been poorly studied in the past, it is shown that eastern Palearctic specimens identified by previous authors as either of the two species in fact belongs to a third species. The synonymized name Graphoderus elatus Sharp, 1882, is reinstated as a valid species (stat. n.) and a lectotype is designated from the mixed syntype series. The male genitalia of all known Graphoderus species have been examined and an illustrated identification key to the genus is provided. The three species in the complex of focus, Graphoderus elatus, Graphoderus zonatus and Graphoderus perplexus are found to have allopatric distributions; Graphoderus perplexus in the Nearctic region, Graphoderus zonatus in the west Palearctic region and eastwards to the Yenisei-Angara river and Graphoderus elatus east of the Yenisei-Angara river. All previous records of either Graphoderus zonatus or Graphoderus perplexus in the east Palearctic, east of the Yenisei-Angara river turned out to be misidentified Graphoderus elatus. This conclusion also brings with it that dimorphic females, thought only to be present in the single subspecies Graphoderus zonatus verrucifer (CR Sahlberg, 1824), proved to be present also in a second species, Graphoderus elatus. The dimorphic female forms is either with dorsally smooth elytra and pronotum or conspicuously granulated elytra and wrinkly pronotum. As has been shown in Graphoderus zonatus verrucifer there is a correlation between the occurrence of granulate female forms in a population and an increase in the number of adhesive discs on pro- and mesotarsus in males within Graphoderus elatus.


Frequent daytime naps predict vocabulary growth in early childhood.
The facilitating role of sleep for language learning is well-attested in adults and to a lesser extent in infants and toddlers. However, the longitudinal relationship between sleep patterns and early vocabulary development is not well understood. This study investigates how measures of sleep are related to the development of vocabulary size in infants and toddlers. Day and night-time sleeping patterns of infants and toddlers were compared with their concurrent and subsequent vocabulary development. Sleep assessments were conducted using a sleep diary specifically designed to facilitate accurate parental report. Sleep measures were used as predictors in a multilevel growth curve analysis of vocabulary development. The number of daytime naps was positively associated with both predicted expressive (p = .062) and receptive vocabulary growth (p = .006), whereas the length of night-time sleep was negatively associated with rate of predicted expressive vocabulary growth (p = .045). Sleep efficiency was also positively associated with both predicted receptive (p = .001) and expressive vocabulary growth (p = .068). These results point to a longitudinal relationship between sleep and language development, with a particular emphasis on the importance of napping at this age.


[External performance evaluation of screening in blood banks in Argentina: results and strategies for improvement].
Because there was no program for the external evaluation of the quality of the screening for transfusion-transmitted infections (TTIs) in blood banks in Argentina, in 1999 the Hemotherapy Service of Garrahan Hospital, in Buenos Aires, launched an ongoing external evaluation program, with the support of the Pan American Health Organization and the Argentine Hemotherapy and Immunohematology Association. A panel of 12 samples that were reactive to all the markers screened for in Argentine blood banks was distributed. The panel was delivered to 52 laboratories in 1999, 102 laboratories in 2000, and 118 laboratories in 2001. The participating laboratories were classified into one of four categories according to their performance: A: 0 or 1 false positive (FP) results; B: 2 or 3 FPs; C: 4 or more FPs; and D: at least 1 false negative result. Workshops were held with the participants in order to jointly analyze the results. Out of the laboratories that received the panel of samples, the percentage of laboratories that sent in their results was 92.3% in 1999, 92.2% in 2000, and 83.9% in 2001. These response levels demonstrate the interest in evaluation activities. However, the annual average percentage of the laboratories that received a "D" classification was very high (30%), which indicates that there are problems in the performance of TTI screening. This poor showing could be related to the difficulty in taking corrective measures throughout the system, given the large number of laboratories involved. Evaluations of this type should be included in Argentina's National Blood Program as a tool for the ongoing assessment of the blood banking situation, for decision-making, and for monitoring the quality of screening.