Macrophage colony-stimulating factor receptor marks and regulates a fetal myeloid-primed B-cell progenitor in mice.
Although it is well established that unique B-cell lineages develop through distinct regulatory mechanisms during embryonic development, much less is understood about the differences between embryonic and adult B-cell progenitor cells, likely to underpin the genetics and biology of infant and childhood PreB acute lymphoblastic leukemia (PreB-ALL), initiated by distinct leukemia-initiating translocations during embryonic development. Herein, we establish that a distinct subset of the earliest CD19(+) B-cell progenitors emerging in the E13.5 mouse fetal liver express the colony-stimulating factor-1 receptor (CSF1R), previously thought to be expressed, and play a lineage-restricted role in development of myeloid lineages, and macrophages in particular. These early embryonic CSF1R(+)CD19(+) ProB cells also express multiple other myeloid genes and, in line with this, possess residual myeloid as well as B-cell, but not T-cell lineage potential. Notably, these CSF1R(+) myeloid-primed ProB cells are uniquely present in a narrow window of embryonic fetal liver hematopoiesis and do not persist in adult bone marrow. Moreover, analysis of CSF1R-deficient mice establishes a distinct role of CSF1R in fetal B-lymphopoiesis. CSF1R(+) myeloid-primed embryonic ProB cells are relevant for infant and childhood PreB-ALLs, which frequently have a bi-phenotypic B-myeloid phenotype, and in which CSF1R-rearrangements have recently been reported.


Evaluation of a soluble fibrin assay in patients with suspected deep vein thrombosis.
The objective of this study was to determine the clinical utility of an enzyme immunoassay (EIA) for soluble fibrin in patients with clinically suspected deep vein thrombosis (DVT). 101 unselected patients with clinically suspected DVT underwent blood sampling for measurement of plasma levels of soluble fibrin, and objective testing for DVT. According to results of the objective tests, patients were classified as DVT-positive (n = 34) or DVT-negative (n = 67). Using different cut-points of soluble fibrin results, the sensitivities, specificities, positive and negative predictive values of the soluble fibrin assay were calculated. A soluble fibrin result of < or = 0.75 mg/ml showed a sensitivity and negative predictive value of 100%, and a specificity of 17.9% for DVT, a soluble fibrin result of < or = 1.40 mg/ml showed a sensitivity of 91.2% and a negative predictive value of 93.6%, and a specificity of 65.7% for DVT, whereas a soluble fibrin result of < or = 8.0 mg/ml showed a specificity and positive predictive value of 100% for DVT. This study demonstrates that the soluble fibrin assay used in the study has potential clinical utility as a diagnostic test in patients with clinically suspected DVT and supports further evaluation of this assay.


A survey of quantitative descriptions of molecular structure.
Numerical characterization of molecular structure is a first step in many computational analysis of chemical structure data. These numerical representations, termed descriptors, come in many forms, ranging from simple atom counts and invariants of the molecular graph to distribution of properties, such as charge, across a molecular surface. In this article we first present a broad categorization of descriptors and then describe applications and toolkits that can be employed to evaluate them. We highlight a number of issues surrounding molecular descriptor calculations such as versioning and reproducibility and describe how some toolkits have attempted to address these problems.


Which nucleotides in the "-10" region are crucial to obtain a fully active MalT-dependent promoter?
We have replaced the hexanucleotide corresponding to the "-10" region of malPp, a positively regulated promoter from Escherichia coli, by random nucleotide sequences and isolated 48 different variants that were as active as the wild-type promoter. Analysis of the nucleotide sequence of their "-10" region strongly suggests that the nature of the nucleotide present at three positions plays a crucial role: 46 of the 48 malPp variants contained C or T at position -12, A at position -11 and T at position -7. The nucleotide composition at the three other positions seems to be much more flexible. The features of these "-10" regions are similar to those of the constitutive promoters, but some significant differences are noticeable and will be discussed. In contrast to other positively regulated promoters, none of the various "-10" regions, including the consensus sequence TATAAT, led to a constitutive promoter.


Alpha-tocopherol supplementation decreases the oxidative susceptibility of LDL in renal failure patients on dialysis therapy.
Atherosclerotic cardiovascular disease is the leading cause of death in patients with end stage renal disease (ESRD) who have undergone dialysis treatment. The oxidation of low density lipoprotein (LDL) appears to be a crucial step in the pathogenesis of atherosclerosis. The increased oxidative stress and attendant increased oxidizability of lipoproteins, such as LDL could contribute to the accelerated atherosclerosis in dialysis patients. Since alpha-tocopherol (AT) is the major antioxidant in LDL, the aim of the present study was to test the effectiveness of RRR-AT supplementation (800 I.U. per day) for 12 weeks on the susceptibility of LDL to oxidation. The study subjects comprised patients with chronic renal failure on hemodialysis (HD), peritoneal dialysis (PD), and age and sex matched controls (C). Plasma fatty acids, lipoproteins and AT levels were measured in these subjects before and after supplementation. Also, LDL AT and oxidizability was studied. LDL was isolated by ultracentrifugation at baseline and after 12 weeks of supplementation, and subjected to a 5-h time course of copper catalyzed oxidation. Oxidation was measured by the formation of conjugated dienes (CD) and lipid peroxides (LP). Supplementation with AT did not alter the plasma lipid or lipoprotein profile of these subjects. Plasma lipid-standardized AT and LDL AT concentrations were not different among the groups at baseline. AT supplementation significantly increased plasma lipid-standardized AT (C=150%, HD=149%, PD=217%, P<0.001) and LDL AT concentrations (C=94%, HD=94%, PD=135%, P<0.003). AT enrichment of LDL resulted in a significant prolongation in conjugated diene lag phase in all groups (C=34%, HD=21%, PD=54%, P<0.02). Lipid peroxide lag phase was also increased significantly in C (27%,) and PD (40%) groups after AT supplementation (P<0.01). There was a significant positive correlation between plasma lipid standardized AT and lag phase (r=0. 54, P=0.0003). Overall, AT decreased the susceptibility of LDL to oxidation in patients with chronic renal failure but the benefit appears to be greater in patients on PD. Therefore, AT supplementation may also provide a measure of protection against CAD in patients with chronic renal failure on dialysis therapy.


Genome-wide association study implicates a chromosome 12 risk locus for late-onset Alzheimer disease.
Only Apolipoprotein E polymorphisms have been consistently associated with the risk of late-onset Alzheimer disease (LOAD), but they represent only a minority of the underlying genetic effect. To identify additional LOAD risk loci, we performed a genome-wide association study (GWAS) on 492 LOAD cases and 498 cognitive controls using Illumina's HumanHap550 beadchip. An additional 238 cases and 220 controls were used as a validation data set for single-nucleotide polymorphisms (SNPs) that met genome-wide significance. To validate additional associated SNPs (p < 0.0001) and nominally associated candidate genes, we imputed SNPs from our GWAS using a previously published LOAD GWAS(1) and the IMPUTE program. Association testing was performed with the Cochran-Armitage trend test and logistic regression, and genome-wide significance was determined with the False Discovery Rate-Beta Uniform Mixture method. Extensive quality-control methods were performed at both the sample and the SNP level. The GWAS confirmed the known APOE association and identified association with a 12q13 locus at genome-wide significance; the 12q13 locus was confirmed in our validation data set. Four additional highly associated signals (1q42, 4q28, 6q14, 19q13) were replicated with the use of the imputed data set, and six candidate genes had SNPs with nominal association in both the GWAS and the joint imputated data set. These results help to further define the genetic architecture of LOAD.


Low birth weight-associated adult hypertension in the rat.
Epidemiological surveys have suggested that intrauterine growth retardation is a risk factor for the development of hypertension in later life. A rat model of intrauterine growth retardation, induced by maternal low-protein diet during the second half of pregnancy, was used to study the relationship between birth weight and adult hypertension. The offspring were born at term and were allowed to nurse normally until weaned to standard chow at 4 weeks of age. They had 15% lower birth weights than control offspring, with complete catch-up growth by age 4 weeks. Both females and males developed progressively worsening hypertension beginning at 8 weeks. The 11-month survival rate was 69% versus 100% in control animals. During the early stages of the hypertension, plasma creatinine was normal, plasma sodium concentration was slightly higher than that of control animals, plasma renin activity was suppressed, and the males had mild proteinuria. Renal function remained normal throughout the 11-month observation period, but plasma renin activity gradually rose above control values. Angiotensin-converting enzyme inhibition by enalapril, begun at 8 weeks of age, was effective in completely normalizing the blood pressure, but did not totally prevent the extra mortality. Sprague-Dawley and Wistar rat strains developed equally severe hypertension after maternal protein deprivation, despite their different susceptibilities to nephrosclerosis with aging. In conclusion, maternal low-protein diet resulted in low birth weight and adult hypertension in the rat. Primary sodium retention and expanded extracellular volume may be critical factors during the development of the hypertension.


Canadian pediatricians: demographic characteristics, perceptions of training, and continuing medical education.
A nationwide survey of Canadian pediatricians was undertaken to answer questions about demographic and practice characteristics, perceptions of the quantity and quality of residency training in relation to the realities of practice, and the patterns of use and the value of continuing medical education. The findings included a lower average age of pediatricians from that determined 10 years earlier, a higher proportion of women practising pediatrics, and higher proportions of pediatricians entering practice in smaller communities, doing geographic full-time university work and doing mainly consulting work. Pediatrics is still perceived as an attractive discipline, but there is dissatisfaction with the quantity and quality of training in adolescent medicine, ophthalmology, dermatology, psychosocial pediatrics and orthopedics. The changing patterns of continuing medical education among the most recent certificants suggest a need for journals and professional societies to assess how they can better meet the needs of Canadian pediatricians in this area.


Endoscopic venous valve transplantation with a valve-stent device.
In this report we describe the outcome of a 6-week study of the valve-stent device in a goat model. Six female goats underwent endoscopic subcutaneous harvest of a valve-containing segment of external jugular vein. This segment was sutured inside a 10-mm Wallstent (Schneider Inc., USA), and the device was compressed and loaded into a 12 Fr. introducer sheath and deployed through a venotomy into the contralateral external jugular vein. The animals were anticoagulated for 1 week and euthanized at 6 weeks. Flow studies, manual strip test, and angioscopic evaluation were performed prior to sacrifice. All six valve-stents were patent at harvest by Doppler flow studies, and five of six were competent as demonstrated by the manual strip test. At angioscopic evaluation, the valves were open. Gross and histologic examination revealed intact valve leaflets in five specimens. This technique is a minimally invasive approach to valve harvest and transplantation with a valve-stent device. We have demonstrated that valve-stent can remain patent and competent for up to 6 weeks. This technique may have potential application in venous valvular incompetence.


Solvent switchable nanostructures and the function of a π-amphiphile.
This manuscript reports solvent tunable functional nano-assemblies of an unsymmetrical bola-shaped π-amphiphile (NDI-PY) which consists of a hydrophobic naphthalene-diimide (NDI) chromophore connected to a non-ionic hydrophilic wedge and a pyridine group at its two opposite arms. Importantly, it contains a hydrazide group located at the hydrophobic domain between the NDI-chromophore and the hydrophilic-wedge to drive the supramolecular assembly by directional H-bonding. NDI-PY exhibits spontaneous assembly in water as well as in a highly non-polar solvent like tetra-chloroethylene (TCE) by the synergistic effect of H-bonding and π-stacking interaction. Spectroscopy studies reveal almost identical self-assembly features in water and TCE with critical aggregation concentrations in the range of 0.3 mM, which matches the values obtained from the isothermal calorimetry (ITC) dilution experiment. Differential scanning calorimetry (DSC) experiments reveal a single endothermic peak at 31 °C (ΔH = -12.3 kJ mol-1) and 40 °C (ΔH = -5.35 kJ mol-1) for water and TCE, respectively, indicating marginally higher thermal stability in TCE, which is consistent with the FT-IR data, suggesting stronger H-bonding in TCE. Although the molecular assembly features appear to be similar, NDI-PY produces distinctly different mesoscopic structures in water and TCE. In water, it forms vesicles (Dh = 150-180 nm) with the pyridine groups displayed at the outer surface, while in TCE it generates a transparent gel (CGC = 8.0 mM) with a nanotubular (width ∼50 nm, wall thickness ∼10 nm) morphology. Powder X-ray diffraction studies show clearly different packing structures; in water a single sharp peak at the low angle (d = 19.3 Å, a little shorter than the extended length of the molecule) suggests the formation of a monolayer membrane, while in TCE several sharp peaks appear with the d values maintaining a ratio of 1 : 1/√3 : 1/2 : 1/√7 : 1/3 : 1/√12, indicating the formation of a 2D hexagonal lattice. Photoconductivity measurements reveal moderate electronic conduction in both cases. However, it shows a remarkable enhancement of the life time of the charge-carriers for the nanotubular structure compared to the vesicular morphology. On the other hand, the vesicles in water display antimicrobial activity against Gram-positive S. aureus with a highly promising MICLB value of 29.4 μg mL-1. In contrast, it does not lyse human red blood cells even at as high a concentration as 2.5 mg mL-1 (HC50 > 2.5 mg mL-1), implying high selectivity of the NDI-PY vesicles towards bacterial cells over mammalian cells. Display of the pyridine groups at the outer surface of the membrane enables molecular recognition by complementary H-bonding with a carboxylic acid group and thereby facilitates uptake of the attached pyrene chromophores in the NDI-membrane by charge-transfer interaction between the NDI acceptor and the pyrene donor. In fact a Job's plot experiment reveals maximum uptake at a 1 : 1 ratio of the NDI-PY and the pyrene guest, indicating all the pyridine groups are accessible at the vesicular surface.


Heralded single-phonon preparation, storage, and readout in cavity optomechanics.
We show how to use the radiation pressure optomechanical coupling between a mechanical oscillator and an optical cavity field to generate in a heralded way a single quantum of mechanical motion (a Fock state). Starting with the oscillator close to its ground state, a laser pumping the upper motional sideband produces correlated photon-phonon pairs via optomechanical parametric down-conversion. Subsequent detection of a single scattered Stokes photon projects the macroscopic oscillator into a single-phonon Fock state. The nonclassical nature of this mechanical state can be demonstrated by applying a readout laser on the lower sideband to map the phononic state to a photonic mode and performing an autocorrelation measurement. Our approach proves the relevance of cavity optomechanics as an enabling quantum technology.


The Item versus the Object in Memory: On the Implausibility of Overwriting As a Mechanism for Forgetting in Short-Term Memory.
The nature of forgetting in short-term memory remains a disputed topic, with much debate focussed upon whether decay plays a fundamental role (Berman et al., 2009; Altmann and Schunn, 2012; Barrouillet et al., 2012; Neath and Brown, 2012; Oberauer and Lewandowsky, 2013; Ricker et al., 2014) but much less focus on other plausible mechanisms. One such mechanism of long-standing in auditory memory is overwriting (e.g., Crowder and Morton, 1969) in which some aspects of a representation are "overwritten" and rendered inaccessible by the subsequent presentation of a further item. Here, we review the evidence for different forms of overwriting (at the feature and item levels) and examine the plausibility of this mechanism both as a form of auditory memory and when viewed in the context of a larger hearing, speech and language understanding system.


Why are there no great women chefs?
This article applies the rhetorical and deliberately provocative approach of the watershed essay art historian Linda Nochlin wrote in 1971—"Why Have there Been No Great Women Artists?"—to today's culinary industry. Nochlin used the question her title posed as a theoretical trap that would draw attention not only to the inherent sexism or prejudice that pervades the way the public perceives art, but also to those same issues' existence within and impact on academia and the other cultural institutions responsible for posing these sorts of questions. Nochlin bypassed the obvious and irrelevant debate over women's being less or differently talented and, in so doing, exposed that debate for being a distraction from the heart of the matter: how, sociologically (media) or institutionally (museums, foundations, etc.), people define a "great artist." Although it's 40 years later, the polemic is as effective when used to understand the gender divide in the food world.


Cost-effectiveness of screening for carotid stenosis in asymptomatic persons.
The Asymptomatic Carotid Atherosclerosis Study (ACAS) showed that carotid endarterectomy was beneficial for symptom-free patients with carotid stenosis of 60% or more. This finding raises the question of whether widespread screening to identify cases of asymptomatic carotid stenosis should be implemented. To determine whether a screening program to identify cases of asymptomatic carotid stenosis would be a cost-effective strategy for stroke prevention. Cost-effectiveness analysis using published data from clinical trials. General population of asymptomatic 65-year-old men. Patients who were screened for carotid disease with duplex Doppler ultrasonography were compared with patients who were not screened. If ultrasonography found significant carotid stenosis (> or = 60%), disease was confirmed by angiography before carotid endarterectomy was done. Quality-adjusted life-years, costs, and marginal cost-effectiveness ratios. When the conditions and results of ACAS were modeled and it was assumed that the survival advantage produced by endarterectomy would last for 30 years, the lifetime marginal cost-effectiveness of screening relative to no screening was $120,000 per quality-adjusted life-year. Sensitivity analysis showed that marginal cost-effectiveness decreased to $50,000 or less per quality-adjusted life-year only under implausible conditions (for example, if a free screening instrument with perfect test characteristics was used or an asymptomatic population with a 40% prevalence of carotid stenosis was found). Surgery offers a real but modest absolute reduction in the rate of stroke at a substantial cost. A program to identify candidates for endarterectomy by screening asymptomatic populations for carotid stenosis costs more per quality-adjusted life-year than is usually considered acceptable.


Changes in ocular aquaporin expression following optic nerve crush.
Changes in the expression of water channels (aquaporins; AQP) have been reported in several diseases. However, such changes and mechanisms remain to be evaluated for retinal injury after optic nerve crush (ONC). This study was designed to analyze changes in the expression of AQP4 (water selective channel) and AQP9 (water and lactate channel) following ONC in the rat. Rat retinal ganglion cells (RGCs) were retrogradely labeled by applying FluoroGold onto the left superior colliculus 1 week before ONC. Retinal injuries were induced by ONC unilaterally. Real-time PCR was used to measure changes in AQP4, AQP9, thy-1, Kir4.1 (K(+) channel), and beta-actin messages. Changes in AQP4, AQP9, Kir4.1, B cell lymphoma-x (bcl-xl), and glial fibrillary acidic protein (GFAP) expression were measured in total retinal extracts using western blotting. The number of RGCs labeled retrogradely from the superior colliculus was 2,090+/-85 cells/mm(2) in rats without any treatment, which decreased to 1,091+/-78 (47% loss) and 497+/-87 cells/mm(2) (76% loss) on days 7 and 14, respectively. AQP4, Kir4.1, and thy-1 protein levels decreased at days 2, 7, and 14, which paralleled a similar reduction in mRNA levels, with the exception of Kir4.1 mRNA at day 2 showing an apparent upregulation. In contrast, AQP9 mRNA and protein levels showed opposite changes to those observed for the latter targets. Whereas AQP9 mRNA increased at days 2 and 14, protein levels decreased at both time points. AQP9 mRNA decreased at day 7, while protein levels increased. GFAP (a marker of astrogliosis) remained upregulated at days 2, 7, and 14, while bcl-xl (anti-apoptotic) decreased. The reduced expression of AQP4 and Kir4.1 suggests dysfunctional ion coupling in retina following ONC and likely impaired retinal function. The sustained increase in GFAP indicates astrogliosis, while the decreased bcl-xl protein level suggests a commitment to cellular death, as clearly shown by the reduction in the RGC population and decreased thy-1 expression. Changes in AQP9 expression suggest a contribution of the channel to retinal ganglion cell death and response of distinct amacrine cells known to express AQP9 following traumatic injuries.


Mapping of adult plant stripe rust resistance genes in diploid A genome wheat species and their transfer to bread wheat.
Stripe rust, caused by Puccinia striiformis West. f.sp. tritici, is one of the most damaging diseases of wheat worldwide. Forty genes for stripe rust resistance have been catalogued so far, but the majority of them are not effective against emerging pathotypes. Triticum monococcum and T. boeoticum have excellent levels of resistance to rusts, but so far, no stripe rust resistance gene has been identified or transferred from these species. A set of 121 RILs generated from a cross involving T. monococcum (acc. pau14087) and T. boeoticum (acc. pau5088) was screened for 3 years against a mixture of pathotypes under field conditions. The parental accessions were susceptible to all the prevalent pathotypes at the seedling stage, but resistant at the adult plant stage. Genetic analysis of the RIL population revealed the presence of two genes for stripe rust resistance, with one gene each being contributed by each of the parental lines. A linkage map with 169 SSR and RFLP loci generated from a set of 93 RILs was used for mapping these resistance genes. Based on phenotypic data for 3 years and the pooled data, two QTLs, one each in T. monococcum acc. pau14087 and T. boeoticum acc. pau5088, were detected for resistance in the RIL population. The QTL in T. monococcum mapped on chromosome 2A in a 3.6 cM interval between Xwmc407 and Xwmc170, whereas the QTL from T. boeoticum mapped on 5A in 8.9 cM interval between Xbarc151 and Xcfd12 and these were designated as QYrtm.pau-2A and QYrtb.pau-5A, respectively. Based on field data for 3 years, their R2 values were 14 and 24%, respectively. T. monococcum acc. pau14087 and three resistant RILs were crossed to hexaploid wheat cvs WL711 and PBW343, using T. durum as a bridging species with the objective of transferring these genes into hexaploid wheat. The B genome of T. durum suppressed resistance in the F1 plants, but with subsequent backcrossing one resistance gene could be transferred from one of the RILs to the hexaploid wheat background. This gene was derived from T. boeoticum acc. pau5088 as indicated by co-introgression of T. boeoticum sequences linked to stripe rust resistance QTL, QYrtb.pau-5A. Homozygous resistant progenies with 40-42 chromosomes have been identified.


Home-ownership in Europe: How did it happen?
We use data from the third wave of the Survey of Health, Ageing and Retirement in Europe (SHARELIFE)(1) to document the different ways individuals first became home-owners across countries and over cohorts over the second half of the 20th century. Focusing on first-time home owners we find that younger cohorts became home-owners earlier and were more likely to do it through credit, less likely to inherit their home directly. Having higher human capital, being employed, married, having children and living in an urban area, all make it more likely to purchase a home with a mortgage. The persistence of family help in accessing home-ownership in many countries demonstrates the interrelation between family, market and the state in most of continental Europe.


CD32a is a marker of a CD4 T-cell HIV reservoir harbouring replication-competent proviruses.
The persistence of the HIV reservoir in infected individuals is a major obstacle to the development of a cure for HIV. Here, using an in vitro model of HIV-infected quiescent CD4 T cells, we reveal a gene expression signature of 103 upregulated genes that are specific for latently infected cells, including genes for 16 transmembrane proteins. In vitro screening for surface expression in HIV-infected quiescent CD4 T cells shows that the low-affinity receptor for the immunoglobulin G Fc fragment, CD32a, is the most highly induced, with no detectable expression in bystander cells. Notably, productive HIV-1 infection of T-cell-receptor-stimulated CD4 T cells is not associated with CD32a expression, suggesting that a quiescence-dependent mechanism is required for its induction. Using blood samples from HIV-1-positive participants receiving suppressive antiretroviral therapy, we identify a subpopulation of 0.012% of CD4 T cells that express CD32a and host up to three copies of HIV DNA per cell. This CD32a+ reservoir was highly enriched in inducible replication-competent proviruses and can be predominant in some participants. Our discovery that CD32a+ lymphocytes represent the elusive HIV-1 reservoir may lead to insights that will facilitate the specific targeting and elimination of this reservoir.


Development of community strategies supporting brief alcohol advice in three Latin American countries: a protocol.
Brief alcohol advice offered to patients was shown to be a clinically- and cost-effective intervention to prevent and manage alcohol-related health harm. However, this intervention is not yet optimally implemented in practice. A suggested strategy to improve the implementation of brief alcohol advice is through community actions which would enhance the environment in which primary healthcare providers must deliver the intervention. However, there has been scarce research conducted to date regarding which community actions have most influence on the adoption and implementation of brief alcohol advice. The current protocol presents the development of a package of community actions to be implemented in three Latin American municipalities, in Colombia, Mexico and Peru. The community actions were based on the Institute for Health Care Improvement's framework for going to full scale, and include: (i) involvement of a Community Advisory Board, (ii) involvement of a project champion, (iii) adoption mechanisms, (iv) support systems and (v) a communication campaign. By presenting a protocol for developing community actions with input from local stakeholders, this article contributes to advancing the public health field of alcohol prevention by potentially stimulating the sustainable adoption and implementation of brief alcohol advice in routine practice.


A Deep-Sequencing Workflow for the Fast and Efficient Generation of High-Quality African Swine Fever Virus Whole-Genome Sequences.
African swine fever (ASF) is a severe disease of suids caused by African swine fever virus (ASFV). Its dsDNA genome (170-194 kbp) is scattered with homopolymers and repeats as well as inverted-terminal-repeats (ITR), which hamper whole-genome sequencing. To date, only a few genome sequences have been published and only for some are data on sequence quality available enabling in-depth investigations. Especially in Europe and Asia, where ASFV has continuously spread since its introduction into Georgia in 2007, a very low genetic variability of the circulating ASFV-strains was reported. Therefore, only whole-genome sequences can serve as a basis for detailed virus comparisons. Here, we report an effective workflow, combining target enrichment, Illumina and Nanopore sequencing for ASFV whole-genome sequencing. Following this approach, we generated an improved high-quality ASFV Georgia 2007/1 whole-genome sequence leading to the correction of 71 sequencing errors and the addition of 956 and 231 bp at the respective ITRs. This genome, derived from the primary outbreak in 2007, can now serve as a reference for future whole-genome analyses of related ASFV strains and molecular approaches. Using both workflow and the reference genome, we generated the first ASFV-whole-genome sequence from Moldova, expanding the sequence knowledge from Eastern Europe.


Impact ejecta horizon within late precambrian shales, adelaide geosyncline, South australia.
A solitary layer of shattered crustal rock fragments has been traced over a distance of 260 kilometers within folded 600-million-year-old Precambrian marine shales of the Adelaide Geosyncline, South Australia. The fragments consist entirely of acid to intermediate volcanics (approximately 1575 million years old) displaying shattered mineral grains, shock lamellae in quartz, and small shatter cones. Fragments reach 30 centimeters in diameter and show evidence of vertical fall emplacement. Available evidence points to derivation of the rock fragments from a distant hypervelocity impact into the Gawler Range Volcanics at Lake Acraman, approximately 300 kilometers west of the Adelaide Geosyncline.


A Comparison of Children's Physical Activity Levels in Physical Education, Recess, and Exergaming.
To compare young children's different intensity physical activity (PA) levels in physical education, recess and exergaming programs. Participants were 140 first and second grade children (73 girls; Meanage= 7.88 years). Beyond the daily 20-minute recess, participants attended 75-minute weekly physical education classes and another 75-minute weekly exergaming classes. Children's PA levels were assessed by ActiGraph GTX3 accelerometers for 3 sessions in the 3 programs. The outcome variables were percentages of time spent in sedentary, light PA and moderate-to-vigorous PA (MVPA). There were significant main effects for program and grade, and an interaction effect for program by grade. Specifically, children's MVPA in exergaming and recess was higher than in physical education. The 2nd-grade children demonstrated lower sedentary behavior and MVPA than the first-grade children during recess; less light PA in both recess and exergaming than first-grade children; and less sedentary behavior but higher MVPA in exergaming than first-grade children. Young children generated higher PA levels in recess and exergaming as compared with physical education. Hence, other school-based PA programs may serve as essential components of a comprehensive school PA program. Implications are provided for educators and health professionals.


The Correlation between Enrolment Criteria and Students' Success in the Study of Medicine: a Five-Year Retrospective Study.
To investigate whether the students' study-success correlates with the achievements evaluated in the enrolment process based on the State Graduation Exam. The retrospective study included a total of 637 students enrolled in the study of Medicine at the Faculty of Medicine from the academic year 2010-11 to 2014-15. We analysed the structure of students enrolled on the basis of the State Graduation Exam, examined their success in two courses of the study programme and then correlated this study-success with individual admission parameters. In particular, we investigated whether the success in the study was influenced by the changes made in the enrolment requirements during this period. Pearson's correlation coefficient was used to measure strenght of correlation between two variables. The changes in the admission parameters, i.e., the inclusion of STEM subjects as obligatory enrolment requirements, made in 2012-13 enabled a better selection of students with high average secondary education grades and with significantly better results in Biology, Physics and particularly in Chemistry. This resulted in a higher success in the study, especially in the last two new-criteria generations. The Pearson correlation coefficient between students' success in Physiology and Pathophysiology II and the achievements valuated in enrolment procedure as total cumulative score were statistically significant (r=0.27, P<0.001). Total points achieved in enrolment procedure was also correlated with students' success in Medical Physics and Biophysics and correlation was even stronger (r=0.52, P<0.001). Our results proved that a well-designed combination of the relevant admission parameters is a crucial prerequisite for a better success in the study.


Emerging Salmonella Enteritidis anaerogenic phage type 14b: outbreak in Norwegian, Swedish and Finnish travellers returning from Greece.
In July 2001, the Norwegian Institute of Public Health (Folkehelseinstituttet, FHI) reported a cluster of Salmonella Enteritidis of phage type 14b infections in Norwegian travellers returning from Greece. An increase in the same uncommon phage type was also registered in Sweden and Finland at the same time. Cases of S. Enteritidis PT 14b in patients returning from Greece were reported in these three Nordic countries in 2001 (303 cases), 2002 (164 cases) and 2003 (199 cases). Case-control studies performed in 2001 in Norway and Sweden indicated that consumption of chicken was associated with illness. In 2002 and 2003, continuing case reports indicated that this uncommon phage type had probably become established in the Greek food chain. Tour operators were informed and contacts were made with Greek public health authorities. Because place of infection is not systematically included in most Salmonella notification systems, the S. Enteritidis phage type 14b outbreak reported here may represent only part of a larger outbreak among travellers visiting Greece. Infections are often reported only in the tourists' home countries and public health authorities in the tourist destinations may not be aware of the problem. Further collaboration between national institutes of public health in Europe is needed to detect outbreaks occurring among tourists.


Genomic changes associated with adaptation to arid environments in cactophilic Drosophila species.
Insights into the genetic capacities of species to adapt to future climate change can be gained by using comparative genomic and transcriptomic data to reconstruct the genetic changes associated with such adaptations in the past. Here we investigate the genetic changes associated with adaptation to arid environments, specifically climatic extremes and new cactus hosts, through such an analysis of five repleta group Drosophila species. We find disproportionately high rates of gene gains in internal branches in the species' phylogeny where cactus use and subsequently cactus specialisation and high heat and desiccation tolerance evolved. The terminal branch leading to the most heat and desiccation resistant species, Drosophila aldrichi, also shows disproportionately high rates of both gene gains and positive selection. Several Gene Ontology terms related to metabolism were enriched in gene gain events in lineages where cactus use was evolving, while some regulatory and developmental genes were strongly selected in the Drosophila aldrichi branch. Transcriptomic analysis of flies subjected to sublethal heat shocks showed many more downregulation responses to the stress in a heat sensitive versus heat resistant species, confirming the existence of widespread regulatory as well as structural changes in the species' differing adaptations. Gene Ontology terms related to metabolism were enriched in the differentially expressed genes in the resistant species while terms related to stress response were over-represented in the sensitive one. Adaptations to new cactus hosts and hot desiccating environments were associated with periods of accelerated evolutionary change in diverse biochemistries. The hundreds of genes involved suggest adaptations of this sort would be difficult to achieve in the timeframes projected for anthropogenic climate change.


Measuring an individual researcher's impact: new directions and challenges.
Metrics which quantify the impact of a scientist are increasingly incorporated into decisions about how to rate and fund individuals and institutions. Several commonly used metrics, based on journal impact factors and citation counts, have been criticised as they do not reliably predict real-world impact, are highly variable between fields and are vulnerable to gaming. Bibliometrics have been incorporated into systems of research assessment but these may create flawed incentives, failing to reward research that is validated, reproducible and with wider impacts. A recent proposal for a new standardised citation metric based on a composite indicator of 6 measures has led to an online database of 100 000 of the most highly cited scientists in all fields. In this perspective article, we provide an overview and evaluation of this new citation metric as it applies to mental health research. We provide a summary of its findings for psychiatry and psychology, including clustering in certain countries and institutions, and outline some implications for mental health research. We discuss strengths and limitations of this new metric, and how further refinements could align impact metrics more closely with wider goals of scientific research.


Stimulatory and inhibitory effects of interleukin (IL)-4 and IL-13 on the production of cytokines by human peripheral blood mononuclear cells: priming for IL-12 and tumor necrosis factor alpha production.
The production of cytokines in monocytes/macrophages is regulated by several different cytokines that have activating or inhibitory effects. Interleukin (IL)-10, IL-4, IL-13, and transforming growth factor (TGF)-beta are usually considered to be the most important macrophage-deactivating factors, with inhibitory effects on cytokine production. Unlike IL-10 and TGF-beta, which appear to act as downmodulators of many phagocytic cell functions, the mode of action of IL-4 and IL-13 is more complex. Addition of IL-4 and IL-13 to peripheral blood mononuclear cell (PBMC) cultures inhibited production of IL-12, tumor necrosis factor (TNF)-alpha, IL-10, and IL-1 beta induced by lipopolysaccharide (LPS) or Staphylococcus aureus added simultaneously with the cytokines. However, pretreatment of PBMC with IL-4 or IL-13 for > or = 20 h enhanced the production of IL-12 and TNF-alpha in response to LPS or S. aureus several fold in these cells; this IL-4-induced priming for the two cytokines was inhibited by anti-IL-4 neutralizing antibodies. IL-4 priming also enhanced the accumulation of IL-12 and TNF-alpha mRNA induced by LPS and S. aureus. The enhanced accumulation of transcripts for the IL-12 p35 and p40 chains by IL-4 priming was reflected in enhanced secretion of both the IL-12 free p40 chain and the p70 heterodimer. These results suggest an unexpected complexity in the regulatory role of IL-4 and IL-13 in immune responses.


Impact of cocoa flavanol intake on age-dependent vascular stiffness in healthy men: a randomized, controlled, double-masked trial.
Increased vascular stiffness, endothelial dysfunction, and isolated systolic hypertension are hallmarks of vascular aging. Regular cocoa flavanol (CF) intake can improve vascular function in healthy young and elderly at-risk individuals. However, the mechanisms underlying CF bioactivity remain largely unknown. We investigated the effects of CF intake on cardiovascular function in healthy young and elderly individuals without history, signs, or symptoms of cardiovascular disease by applying particular focus on functional endpoints relevant to cardiovascular aging. In a randomized, controlled, double-masked, parallel-group dietary intervention trial, 22 young (<35 years) and 20 elderly (50-80 year) healthy, male non-smokers consumed either a CF-containing drink (450 mg CF) or nutrient-matched, CF-free control drink bi-daily for 14 days. The primary endpoint was endothelial function as measured by flow-mediated vasodilation (FMD). Secondary endpoints included cardiac output, vascular stiffness, conductance of conduit and resistance arteries, and perfusion in the microcirculation. Following 2 weeks of CF intake, FMD improved in young (6.1 ± 0.7 vs. 7.6 ± 0.7 %, p < 0.001) and elderly (4.9 ± 0.6 vs. 6.3 ± 0.9 %, p < 0.001). Secondary outcomes demonstrated in both groups that CF intake decreased pulse wave velocity and lowered total peripheral resistance, and increased arteriolar and microvascular vasodilator capacity, red cell deformability, and diastolic blood pressure, while cardiac output remained affected. In the elderly, baseline systolic blood pressure was elevated, driven by an arterial-stiffness-related augmentation. CF intake decreased aortic augmentation index (-9 %) and thus systolic blood pressure (-7 mmHg; Clinicaltrials.gov: NCT01639781). CF intake reverses age-related burden of cardiovascular risk in healthy elderly, highlighting the potential of dietary flavanols to maintain cardiovascular health.


Relationship of IgG and IgM autoantibodies to oxidized low density lipoprotein with coronary artery disease and cardiovascular events.
The relationship between autoantibodies to oxidized low density lipoprotein (OxLDL) and coronary artery disease (CAD) remains controversial. IgM and IgG OxLDL autoantibodies to malondialdehyde (MDA)-modified LDL, copper oxidized low density lipoprotein (CuOxLDL), and oxidized cholesterol linoleate (OxCL), as well as apolipoprotein B-100 immune complexes (apoB-ICs), were measured in 504 patients undergoing clinically indicated coronary angiography. Patients were followed for cardiovascular events for a median of 4 years. In univariate analysis, IgM OxLDL autoantibodies and IgM apoB-ICs were inversely associated with the presence of angiographically determined CAD, whereas IgG OxLDL autoantibodies and IgG apoB-ICs were positively associated. In logistic regression analysis, compared with the first quartile, patients in the fourth quartile of IgM OxLDL autoantibodies and apoB-ICs showed a lower probability of angiographically determined CAD (>50% diameter stenosis). Odds ratios and (95% confidence intervals) were as follows: MDA-LDL, 0.51 (0.32-0.82; P = 0.005); CuOxLDL, 0.63 (0.39-1.01; P = 0.05); OxCL, 0.63 (0.39-1.01; P = 0.05); and apoB-IC, 0.55 (0.34-0.88; P = 0.013). These relationships were accentuated in the setting of hypercholesterolemia, with the highest IgM levels showing the lowest risk of CAD for the same level of hypercholesterolemia. Multivariable analysis revealed that neither IgM or IgG OxLDL autoantibodies nor apoB-ICs were independently associated with angiographically determined CAD or cardiovascular events. In conclusion, IgG and IgM OxLDL biomarkers have divergent associations with CAD in univariate analysis but are not independent predictors of CAD or clinical events.


Brain structural imaging correlates of olfactory dysfunction in obsessive-compulsive disorder.
Olfactory dysfunction has been described in obsessive-compulsive disorder (OCD). Brain regions involved in smell processing partially overlap with structures included in the neurobiological models of OCD, although no previous studies have analyzed the neuroanatomical correlates of olfactory dysfunction in this disorder. The aim of our study was to examine the association between regional gray matter volume, as assessed by a voxel-based morphometry analysis of magnetic resonance images (MRI), and olfactory function, as assessed by the Sniffin' Sticks test (SST). Olfactory function was assessed in 19 OCD patients and 19 healthy volunteers. All participants were also scanned in a 1.5-T magnet to obtain T1-weighted anatomical MRIs, which were pre-processed and analyzed with SPM8. Three different correlation models were used to study the association between regional gray matter volumes and olfactory function in the domains assessed by the SST: detection threshold, discrimination, and identification. OCD patients showed a significant impairment in all the domains assessed by the SST. Voxel-based mapping revealed a positive association in healthy controls between detection threshold and the gray matter content of a left anterior cingulate cortex cluster. In OCD patients, a positive correlation was observed between identification errors and the gray matter volume of the left medial orbital gyrus. In a post hoc analysis, these two gray matter regions were shown to be enlarged in OCD patients. Our findings support the idea that olfactory dysfunction in OCD is associated with volumetric changes in brain areas typically implicated in the neurobiology of the disorder.


High-dose short-term folate administration modifies ambulatory blood pressure in postmenopausal women. A placebo-controlled study.
In humans low intake or low levels of folate are related to elevated homocysteine (Hcy) or blood pressure (BP). This study was performed to test whether in healthy postmenopausal women a 3-week administration of folate (5-methyltetrahydrofolate: 5-MTHF) at the dose of 15 mg/day (n=15), was, in comparison to placebo (n=15), capable of modifying 24-h ambulatory BP, along with Hcy and insulin metabolism. Placebo did not modify any parameter. 5-MTHF significantly decreased nocturnal systolic (-4.48+/-1.8 mm Hg; P=0.029), diastolic (-5.33+/-1.3 mm Hg; P=0.001) and mean (-5.10+/-1.1 mm Hg; P=0.005) BP, in a way that was significantly different from that observed during placebo. 5-MTHF also reduced Hcy (11.77+/-1.15 vs 8.71+/-0.50 micromol/l; P=0.03), and insulin resistance, evaluated by HOMA-IR (2.58+/-0.04 vs 2.03+/-0.04; P=0.01). Whether maintained in the long term, 5-MTHF's cardiovascular and metabolic effect may contribute to primary cardiovascular prevention of postmenopausal women.


Selective mobilization of cytotoxic leukocytes by epinephrine.
It is well-known that acute stress, presumably as a first defense against pathogens, enhances PBMC counts by mobilizing these beta2-adrenoceptor positive cells from the marginal pool. Yet, only select leukocyte subsets participate in this phenomenon of adrenergic leukocytosis and underlying mechanisms are obscure. In this study, we analyzed in human blood adhesion molecule and chemokine receptor profiles in 14 leukocyte subsets, and responsiveness of subsets to epinephrine in vivo and in vitro. Five subsets, namely, CCR7(-)CD45RA(+)CD8(+) effector T cells, CD4(-)CD8(-) gamma/delta T cells, CD3(+)CD56(+) NKT-like cells, CD16(+)CD56(dim) cytotoxic NK cells, and CD14(dim)CD16(+) proinflammatory monocytes showed a rapid and transient increase after infusion of epinephrine at physiological concentrations. These cells were characterized by a CD62L(-)CD11a(bright)CX3CR(bright) phenotype, whereby expression of both CD11a and CX3CR1 was strongly correlated with adrenergic leukocytosis in vivo (r = 0.86 and 0.78, p < 0.005). The same subsets showed highest adherence to activated endothelium in vitro, which (except for proinflammatory monocytes) was reversed by epinephrine. We conclude that these five cytotoxic effector leukocyte subsets comprise the marginal pool by a CD11a/CX3CR1-mediated attachment to the endothelium. Epinephrine rapidly attenuates this attachment to allow demargination and release of the cells into the circulation that, because of their cytotoxic effector function, provide immediate protection from invading pathogens.


The politics of assessment: water and sanitation MDGs in the Middle East.
The Middle East and North Africa (MENA) is generally considered to be making adequate progress towards meeting Target 10 of the Millennium Development Goals (MDGs), which calls for halving the proportion of the population with inadequate access to drinking water and sanitation. Progress towards achieving Target 10 is evaluated by the Joint Monitoring Programme (JMP), run by UNICEF and WHO. This article shows that the assessment methodologies employed by the JMP significantly overstate coverage rates in the drinking water and sanitation sectors, by overlooking and 'not counting' problems of access, affordability, quality of service and pollution. The authors show that states in MENA often fail to provide safe drinking water and adequate sanitation services, particularly in densely populated informal settlements, and that many centralized water and sanitation infrastructures contribute to water pollution and contamination. Despite the glaring gap between the MDG statistics and the evidence available from national and local reports, exclusionary political regimes in the region have had few incentives to adopt more accurate assessments and improve the quality of service. While international organizations have proposed some reforms, they too lack incentives to employ adequate measures that gauge access, quality and affordability of drinking water and sanitation services.


CdS/ZnS core-shell nanocrystal photosensitizers for visible to UV upconversion.
Herein we report the first example of nanocrystal (NC) sensitized triplet-triplet annihilation based photon upconversion from the visible to ultraviolet (vis-to-UV). Many photocatalyzed reactions, such as water splitting, require UV photons in order to function efficiently. Upconversion is one possible means of extending the usable range of photons into the visible. Vis-to-UV upconversion is achieved with CdS/ZnS core-shell NCs as the sensitizer and 2,5-diphenyloxazole (PPO) as annihilator and emitter. The ZnS shell was crucial in order to achieve any appreciable upconversion. From time resolved photoluminescence and transient absorption measurements we conclude that the ZnS shell affects the NC and triplet energy transfer (TET) from NC to PPO in two distinct ways. Upon ZnS growth the surface traps are passivated thus increasing the TET. The shell, however, also acts as a tunneling barrier for TET, reducing the efficiency. This leads to an optimal shell thickness where the upconversion quantum yield (Φ'UC) is maximized. Here the maximum Φ'UC was determined to be 5.2 ± 0.5% for 4 monolayers of ZnS shell on CdS NCs.


Peptide growth factors signal differentially through protein kinase C to extracellular signal-regulated kinases in neonatal cardiomyocytes.
The extracellular signal-regulated kinases 1/2 (ERK1/2) are activated in cardiomyocytes by Gq protein-coupled receptors and are associated with induction of hypertrophy. Here, we demonstrate that, in primary cardiomyocyte cultures, ERK1/2 were also significantly activated by platelet-derived growth factor (PDGF), epidermal growth factor (EGF) or fibroblast growth factor (FGF), but insulin, insulin-like growth factor 1 (IGF-1) and nerve growth factor (NGF) had relatively minor effects. PDGF, EGF or FGF increased cardiomyocyte size via ERK1/2, whereas insulin, IGF-1 or NGF had no effect suggesting minimum thresholds/durations of ERK1/2 signaling are required for the morphological changes associated with hypertrophy. Peptide growth factors are widely accepted to activate phospholipase C gamma1 (PLCgamma1) and protein kinase C (PKC). In cardiomyocytes, only PDGF stimulated tyrosine phosphorylation of PLCgamma1 and nPKCdelta. Furthermore, activation of ERK1/2 by PDGF, but not EGF, required PKC activity. In contrast, EGF substantially increased Ras.GTP with rapid activation of c-Raf, whereas stimulation of Ras.GTP loading by PDGF was minimal and activation of c-Raf was delayed. Our data provide clear evidence for differential coupling of PDGF and EGF receptors to the ERK1/2 cascade, and indicate that a minimum threshold/duration of ERK1/2 signaling is required for the development of cardiomyocyte hypertrophy.


Analysis of the microcrystalline inclusion compounds of triclosan with β-cyclodextrin and its tris-O-methylated derivative.
Solid 1:1 inclusion compounds of triclosan with native and permethylated β-cyclodextrin (β-CD and TRIMEB) were prepared by co-crystallisation and co-evaporation, respectively, and studied by FT-IR and (13)C{(1)H} CP/MAS NMR spectroscopies, thermogravimetric analysis, X-ray diffraction and theoretical calculations. Results showed that triclosan inclusion into TRIMEB afforded an amorphous solid, whilst β-CD·triclosan is composed of microcrystals belonging to two different phases. In the phase featuring larger crystals, X-ray diffraction was carried out and the β-CD host units, packing head-to-head in infinite channels, were refined; the geometry for the included but highly disordered triclosan molecules was assessed by theoretical calculations. The bacterial growth inhibitory action of the inclusion compounds was studied in comparison to that of pure triclosan on Gram-negative (Salmonella, Escherichia) and Gram-positive strains (Bacillus, Listeria, Enterococcus and Staphylococcus) typically associated with human pathologies, and also on environmental bacteria isolated from different soil and water sources. The antimicrobial activities obtained in the present work showed that, of the two CD hosts, TRIMEB brings the most favourable carrier effect: it reduced the toxicity of triclosan against some of the environmental strains and afforded slightly higher action against virulent strains.


A highly annotated whole-genome sequence of a Korean individual.
Recent advances in sequencing technologies have initiated an era of personal genome sequences. To date, human genome sequences have been reported for individuals with ancestry in three distinct geographical regions: a Yoruba African, two individuals of northwest European origin, and a person from China. Here we provide a highly annotated, whole-genome sequence for a Korean individual, known as AK1. The genome of AK1 was determined by an exacting, combined approach that included whole-genome shotgun sequencing (27.8x coverage), targeted bacterial artificial chromosome sequencing, and high-resolution comparative genomic hybridization using custom microarrays featuring more than 24 million probes. Alignment to the NCBI reference, a composite of several ethnic clades, disclosed nearly 3.45 million single nucleotide polymorphisms (SNPs), including 10,162 non-synonymous SNPs, and 170,202 deletion or insertion polymorphisms (indels). SNP and indel densities were strongly correlated genome-wide. Applying very conservative criteria yielded highly reliable copy number variants for clinical considerations. Potential medical phenotypes were annotated for non-synonymous SNPs, coding domain indels, and structural variants. The integration of several human whole-genome sequences derived from several ethnic groups will assist in understanding genetic ancestry, migration patterns and population bottlenecks.


Acute toxicity of six freshwater mussel species (Glochidia) to six chemicals: implications for Daphnids and Utterbackia imbecillis as surrogates for protection of freshwater mussels (Unionidae).
Acute (24-h) toxicity tests were used in this study to compare lethality responses in early life stages (glochidia) of six freshwater mussel species, Leptodea fragilis, U. imbecillis, Lampsilis cardium, Lampsilis siliquoidea, Megalonaias nervosa, and Ligumia subrostrata, and two standard test organisms, Ceriodaphnia dubia and Daphnia magna. Concentrations of carbaryl, copper, 4-nonylphenol, pentachlorophenol, permethrin, and 2,4-D were used in acute exposures to represent different chemical classes and modes of action. The relative sensitivities of species were evaluated by ranking their LC50 values for each chemical. We used these ranks to determine the extent to which U. imbecillis (one of the most commonly used unionids in toxicity tests) was representative of the tolerances of other mussels. We also calculated geometric mean LC50s for the families Unionidae and Daphnidae. Rankings of these data were used to assess the extent to which Daphnidae can be used as surrogates for freshwater mussels relative to chemical sensitivity. While no single chemical elicited consistently high or low toxicity estimates, carbaryl and 2,4-D were generally the least toxic to all species tested. No species was always the most sensitive, and Daphnidae were generally protective of Unionidae. Utterbackia imbecillis, while often proposed as a standard unionid mussel test species, did not always qualify as a sufficient surrogate (i.e., a substitute organism that often elicits similar sensitivity responses to the same contaminant exposure) for other species of mussels, since it was usually one of the more tolerant species in our rankings. U. imbecillis should be used as a surrogate species only with this caution on its relative insensitivity.


An interdisciplinary consensus on the management of brain metastases in patients with renal cell carcinoma.
Brain metastases are a challenging manifestation of renal cell carcinoma. We have a limited understanding of brain metastasis tumor and immune biology, drivers of resistance to systemic treatment, and their overall poor prognosis. Current data support a multimodal treatment strategy with radiation treatment and/or surgery. Nonetheless, the optimal approach for the management of brain metastases from renal cell carcinoma remains unclear. To improve patient care, the authors sought to standardize practical management strategies. They performed an unstructured literature review and elaborated on the current management strategies through an international group of experts from different disciplines assembled via the network of the International Kidney Cancer Coalition. Experts from different disciplines were administered a survey to answer questions related to current challenges and unmet patient needs. On the basis of the integrated approach of literature review and survey study results, the authors built algorithms for the management of single and multiple brain metastases in patients with renal cell carcinoma. The literature review, consensus statements, and algorithms presented in this report can serve as a framework guiding treatment decisions for patients. CA Cancer J Clin. 2022;72:454-489.


Learning to forget: continual prediction with LSTM.
Long short-term memory (LSTM; Hochreiter & Schmidhuber, 1997) can solve numerous tasks not solvable by previous learning algorithms for recurrent neural networks (RNNs). We identify a weakness of LSTM networks processing continual input streams that are not a priori segmented into subsequences with explicitly marked ends at which the network's internal state could be reset. Without resets, the state may grow indefinitely and eventually cause the network to break down. Our remedy is a novel, adaptive "forget gate" that enables an LSTM cell to learn to reset itself at appropriate times, thus releasing internal resources. We review illustrative benchmark problems on which standard LSTM outperforms other RNN algorithms. All algorithms (including LSTM) fail to solve continual versions of these problems. LSTM with forget gates, however, easily solves them, and in an elegant way.


The selective vulnerability of retinal ganglion cells in rat chronic ocular hypertension model at early phase.
Glutamate neurotoxicity has been postulated to play a prominent role in glaucoma. In this study the possible roles of two subunits of glutamate receptors during the early phase of retinal ganglion cell (RGC) loss in a rat chronic ocular hypertension (COH) model were investigated. COH was induced by applying argon laser to the episcleral and limbal veins of the right eye of rats, the observation times were at 4, 14 and 28 days after the first laser. RGCs were retrogradely labeled by putting Fluoro-Gold (FG) on the surface of both side superior colliculus. Immunohistochemical staining using specific antibodies against N-methyl-D-aspartate receptor 1 (NR1) or glutamate receptor 2/3 (GluR2/3) was performed on the retinal sections of normal and COH eyes. Fluorescent images were captured using confocal laser scanning microscope and the number of NR1 and GluR2/3 labeled cells were counted and cell size was measured using Stereo Investigator. During the observation period, the numbers of NR1 and GluR2/3 positive RGCs in the RGC layer were reduced parallel to the loss of RGC. The dramatic loss of GluR2/3 immunoreactive neurons occurred starting immediately after the first laser to 4 days while the dramatic loss of NR1 immunoreactive neurons occurred from 14 to 28 days after the first laser. Size difference was detected in NR1 immunoreactive RGCs, large ones were more sensitive to the high ocular pressure. These results suggest that both NR1 and GluR2/3 are involved in the mediation of RGC death in the early stage of COH.


Clinical performance of Anyplex II HPV28 by human papillomavirus type and viral load in a referral population.
Anyplex II HPV28 (`Anyplex`) is a semi-quantitative DNA PCR assay divided into set A, comprising 14 high risk (hr)HPV types; and set B, comprising 5 possibly hrHPV types and 9 low risk (lr)HPV types. We compared the ability of Anyplex to that of Hybrid Capture 2 (HC2) and PreTect HPV-Proofer (`Proofer`) to detect cervical intraepithelial neoplasia grade two or worse (CIN2+) by HPV types and viral load. This cross-sectional study included 296 women referred to colposcopy with abnormal cervical cytology and/or persistent HPV infection. CIN2+ was identified in 175/296 women. Liquid based cytology samples were used to perform HPV testing. The sensitivity of Anyplex to detect CIN2+ was 98.9% (95% CI 95.9-99.9) and specificity 43.0% (95% CI 34.0-52.3). Restricting to medium and high viral loads in Anyplex set A, sensitivity and specificity were 97.1% (95% CI 93.5-99.1) and 59.5% (95% CI 50.2-68.3) with positive (PPV) and negative predictive value (NPV) 77.6% and 93.5%, respectively, comparable to HC2. Restricting Anyplex to the hrHPV types in Proofer, HPV16, 18, 31, 33 and 45, sensitivity and specificity for CIN2+ were 85.1% (95% CI 79.0-90.1) and 71.1% (95% CI 62.1-79.0), comparable to Proofer`s. When adding HPV52 and 58, the sensitivity for CIN2+ was 92.6% (95% CI 87.6-96.0) and CIN3+ 96.5% (95% CI 92.0-98.8). No value of Anyplex set B was found in detecting CIN2+. In conclusion, the clinical performance of medium and high viral loads in Anyplex set A was comparable to HC2. Restricting the test to the 7 hrHPV types included in the 9-valent HPV-vaccine, HPV16, 18, 31, 33, 45, 52 and 58, satisfies the international criteria for cervical cancer screening with relative sensitivity compared to HC2 for CIN2+ and CIN3+ of 0.98 and 1.01, respectively. Detecting all 28 Anyplex HPV types adds no benefit in a referral population.


Down-regulation of hepatic lipase gene expression and activity by fenofibrate.
The influence of the hypolipidemic drug, fenofibrate, on hepatic lipase (HL) gene expression and activity was investigated in the rat. Fenofibrate treatment provoked a dose-dependent decrease in HL mRNA levels. At a dose of 0.5% (w/w), HL mRNA levels were reduced to nearly 50% the levels in untreated controls. This decrease was parallelled by a comparable reduction in liver HL activity. The decrease in HL mRNA levels was already observed after 1 day of fenofibrate treatment. Whole liver perfusion experiments showed that the heparin-releasable HL activity in fenofibrate-treated livers dropped to 10% the activity in control livers. In conclusion, treatment with fenofibrate decreases HL gene expression, leading to a lowered activity of endothelium bound HL in fenofibrate-treated livers.


Ablation of insulin-producing neurons in flies: growth and diabetic phenotypes.
In the fruit fly Drosophila, four insulin genes are coexpressed in small clusters of cells [insulin-producing cells (IPCs)] in the brain. Here, we show that ablation of these IPCs causes developmental delay, growth retardation, and elevated carbohydrate levels in larval hemolymph. All of the defects were reversed by ectopic expression of a Drosophila insulin transgene. On the basis of these functional data and the observation that IPCs release insulin into the circulatory system, we conclude that brain IPCs are the main systemic supply of insulin during larval growth. We propose that IPCs and pancreatic islet beta cells are functionally analogous and may have evolved from a common ancestral insulin-producing neuron. Interestingly, the phenotype of flies lacking IPCs includes certain features of diabetes mellitus.


Structure and genetic diversity of Anacardium humile (Anacardiaceae): a tropical shrub.
Anacardium humile Saint Hilaire is a tropical shrub native to the Cerrado biome. It is a fruiting species with biological, medicinal, and socioeconomic significance. Thus, knowing how the genetic variability of natural populations is organized allows for the establishment of strategies for conservation and the sustainable use of the species and its biome. Six microsatellite loci previously developed from Anacardium occidentale were used to investigate the spatial genetic structure and genetic diversity of eight natural A. humile populations based on analyses of 242 adult plants. The results obtained indicate that these populations show a high level of genetic diversity (expected heterozygosity = 0.710). The endogamy coefficient was positive and significant for most populations, with a mean of 0.142 (P = 0.001). The genetic differentiation between populations was low (θ = 0.075 and GST = 0.066) but significant (P = 0.0001). The genotypes of five of the eight populations were non-randomly distributed with clusters of related plants for which the coancestry values were positive and significant. These populations exhibited high and significant endogamy indices. The results obtained for A. humile populations show that genetic conservation programs should be implemented to maintain this species.


Composition of the phyllospheric microbial populations on vegetable plants with different glucosinolate and carotenoid compositions.
The plant phyllosphere is intensely colonized by a complex and highly diverse microbial population and shows pronounced plant-species-specific differences. The mechanisms and influencing factors determining whether and in which density microorganisms colonize plant phyllosphere tissues are not yet fully understood. One of the key influencing factors is thought to be phytochemical concentration and composition. Therefore, correlations between various concentrations of individual glucosinolates and carotenoids in four different plant species-Brassica juncea, Brassica campestris, Cichorium endivia, and Spinacea oleracea-and the phyllospheric bacterial population size associated with the aerial parts of the same plants were analyzed. The concentration of various individual glucosinolates and carotenoids were measured using high-performance liquid chromatography. The phyllospheric bacterial population size including both nonculturable and culturable organisms was assessed using quantitative real-time polymerase chain reaction, and the physiological profile of the culturable microbial community was analyzed using the Biolog system. Results show significant differences between plant species in both concentration and composition of secondary metabolites, bacterial population size, and microbial community composition in three consecutively performed experiments. An interesting and underlying trend was that bacterial density was positively correlated to concentrations of beta-carotene in the plant phyllosphere of the four plant species examined. Likewise, the alkenyl glucosinolates, 2-propenyl, 3-butenyl, and 4-pentenyl, concentrations were positively correlated to the bacterial population density, whereas the aromatic glucosinolate 2-phenylethyl showed a negative correlation to the phyllospheric bacterial population size. Thus, we report for the first time the relationship between individual glucosinolate and carotenoid concentrations and the phyllospheric bacterial population size of nonculturable and culturable organisms and the phyllospheric microbial physiological profiles.


Radiotheranostics - Precision Medicine in Nuclear Medicine and Molecular Imaging.
'See what you treat and treat what you see, at a molecular level', could be the motto of theranostics. The concept implies diagnosis (imaging) and treatment of cells (usually cancer) using the same molecule, thus guaranteeing a targeted cytotoxic approach of the imaged tumor cells while sparing healthy tissues. As the brilliant late Sam Gambhir would say, the imaging agent acts like a 'molecular spy' and reveals where the tumoral cells are located and the extent of disease burden (diagnosis). For treatment, the same 'molecular spy' docks to the same tumor cells, this time delivering cytotoxic doses of radiation (treatment). This duality represents the concept of a 'theranostic pair', which follows the scope and fundamental principles of targeted precision and personalized medicine. Although the term theranostic was noted in medical literature in the early 2000s, the principle is not at all new to nuclear medicine. The first example of theranostic dates back to 1941 when Dr. Saul Hertz first applied radioiodine for radionuclide treatment of thyroid cells in patients with hyperthyroidism. Ever since, theranostics has been an integral element of nuclear medicine and molecular imaging. The more we understand tumor biology and molecular pathology of carcinogenesis, including specific mutations and receptor expression profiles, the more specific these 'molecular spies' can be developed for diagnostic molecular imaging and subsequent radionuclide targeted therapy (radiotheranostics). The appropriate selection of the diagnostic and therapeutic radionuclide for the 'theranostic pair' is critical and takes into account not only the type of cytotoxic radiation emission, but also the linear energy transfer (LET), and the physical half-lives. Advances in radiochemistry and radiopharmacy with new radiolabeling techniques and chelators are revolutionizing the field. The landscape of cytotoxic systemic radionuclide treatments has dramatically expanded through the past decades thanks to all these advancements. This article discusses present and promising future theranostic applications for various types of diseases such as thyroid disorders, neuroendocrine tumors (NET), pediatric malignancies, and prostate cancer (PC), and provides an outlook for future perspectives.


VapCs of Mycobacterium tuberculosis cleave RNAs essential for translation.
The major human pathogen Mycobacterium tuberculosis can survive in the host organism for decades without causing symptoms. A large cohort of Toxin-Antitoxin (TA) modules contribute to this persistence. Of these, 48 TA modules belong to the vapBC (virulence associated protein) gene family. VapC toxins are PIN domain endonucleases that, in enterobacteria, inhibit translation by site-specific cleavage of initiator tRNA. In contrast, VapC20 of M. tuberculosis inhibits translation by site-specific cleavage of the universally conserved Sarcin-Ricin loop (SRL) in 23S rRNA. Here we identify the cellular targets of 12 VapCs from M. tuberculosis by applying UV-crosslinking and deep sequencing. Remarkably, these VapCs are all endoribonucleases that cleave RNAs essential for decoding at the ribosomal A-site. Eleven VapCs cleave specific tRNAs while one exhibits SRL cleavage activity. These findings suggest that multiple vapBC modules contribute to the survival of M. tuberculosis in its human host by reducing the level of translation.


Beliefs about voices in voice-hearers: the role of schema functioning.
Evidence is emerging that beliefs about voices are influenced by broader schematic beliefs about the self and others. Similarly, studies indicate that the relationship an individual has with their voice may mirror wider patterns of relating observed in social relationships, which may be influenced by schematic beliefs. This study examined associations between beliefs about voices and self and other schemas. Furthermore, associations between schemas and the perceived relationship between the hearer and their predominant voice were explored. Forty-four voice-hearing participants were recruited across mental health services. Participants completed self-report measures of beliefs about voices, schema functioning, and relating between the hearer and their voice. Dimensions of voice experience, such as frequency and content, were assessed using a clinician-rated scale. Beliefs about voices correlated with negative voice content and schemas. After controlling for negative voice content, schemas were estimated to predict between 1 and 17% of the variance in the six measured beliefs about voices; three of the associations reached statistical significance. Negative-self schema were the strongest predictors of beliefs about voices, whilst positive-self also showed potential relationships. Schemas also correlated with dimensions of relating between the hearer and their voice. In line with previous research, this study provides evidence that schemas, particularly self-schema, may be important in the development of beliefs about voices. This study offers preliminary findings to suggest that schemas are also associated with the perceived relationship between the hearer and their voice.


Knee structural alteration and BMI: a cross-sectional study.
To describe the associations among BMI, knee cartilage morphology, and bone size in adults. A cross-sectional convenience sample of 372 male and female subjects (mean age, 45 years; range, 26 to 61 years) was studied. Knee articular cartilage defect score (0 to 4) and prevalence (defect score of >/=2), volume, and thickness, as well as bone surface area and/or volume, were determined at the patellar, tibial, and femoral sites using T1-weighted fat-saturation magnetic resonance imaging. Height, weight, BMI, and radiographic osteoarthritis were measured by standard protocols. In multivariate analysis in the whole group, BMI was significantly associated with knee cartilage defect scores (beta: +0.016/kg/m(2) to +0.083/kg/m(2), all p < 0.05) and prevalence (odds ratio: 1.05 to 1.12/kg/m(2), all p < 0.05 except for the lateral tibiofemoral compartment). In addition, BMI was negatively associated with patellar cartilage thickness only (beta = -0.021 mm/kg/m(2); p = 0.039) and was positively associated with tibial bone area (medial: beta = +7.1 mm(2)/kg/m(2), p = 0.001; lateral: beta = +3.2 mm(2)/kg/m(2), p = 0.037). Those who were obese also had higher knee cartilage defect severity and prevalence and larger medial tibial bone area but no significant change in cartilage volume or thickness compared with those of normal weight. This study suggests that knee cartilage defects and tibial bone enlargement are the main structural changes associated with increasing BMI particularly in women. Preventing these changes may prevent knee osteoarthritis in overweight and obese subjects.


Root and Basal Stem Rot of Rose Caused by Phytophthora citrophthora in Italy.
Approximately 800 ha of cut flower roses are cultivated for commercial production in Italy. During autumn of 2004 in an experimental greenhouse in western Sicily (southern Italy), 60% of 2-year-old plants of rose cv. Red France on Rosa indica cv. Major rootstock grown in soil showed leaf chlorosis and wilt. A dark brown lesion lined by a water-soaked area was noticeable at the stem base near the soil surface. Root rot was found consistently associated with aboveground symptoms and plants collapsed within 4 months after the appearance of the first symptoms. The same symptoms were observed sporadically on rose plants of the same cultivar during the last 6 years in commercial nurseries in western Sicily. In all cases, a Phytophthora species has been consistently isolated from rotted roots and stems on Phytophthora-selective media. Pure cultures were obtained by single-hypha transfers. The species was identified as Phytophthora citrophthora on the basis of morphological characters and electrophoretic analysis of mycelial proteins on polyacrylamide gel (1). On potato dextrose agar, isolates produced petaloid colonies with optimum growth temperature at 25°C. On V8 agar, mono- and occasionally bipapillate, ovoid to limoniform sporangia, measuring 44 to 55 × 27 to 28 μm, with a mean length/breadth ratio of 1.4:1 were produced. All isolates were heterothallic but did not produce gametangia in dual cultures with P. nicotianae isolates of A1 and A2 mating type. Electrophoretic patterns of total mycelial proteins and four isozyme (acid and alkaline phosphatases, esterase, and malato dehydrogenase) of the isolates from rose were identical to those of reference isolates of P. citrophthora, but clearly distinct from isolates of other heterothallic species with papillate sporangia, including P. capsici, P. nicotianae, P. palmivora, and P. tropicalis. All isolates from rose showed the same electrophoretic profiles. Blast search of rDNA-ITS sequence from PCR-amplified ITS4/ITS6 primers (2) of a representative isolate from rose (IMI 392044) showed 98% homology with a reference isolate of P. citrophthora (GenBank No. EU0000631), thus confirming the identification. Pathogenicity of isolate IMI 392044 was tested on 10 12-month-old plants of rose cv. Red France grafted on R. indica cv. Major transplanted in pots containing a mixture of sphagnum peat moss and sandy loam soil (1:1 vol/vol) infested with 80 g of inoculum per liter of mixture. Inoculum was produced by growing the isolate on wheat kernels. Plants transplanted in pots containing noninfested soil served as controls. Plants were kept in a greenhouse at 22 ± 3°C and watered to soil saturation once a week. Inoculated plants developed symptoms of leaf chlorosis and root and crown rot within 15 to 30 days and wilted within 40 to 80 days after inoculation. Control plants remained healthy. P. citrophthora was consistently reisolated from inoculated plants. Root and basal stem rot of rose may be caused by several Phytophthora spp. and has been reported in various countries of Asia, Europe, and North America (3,4). However, to our knowledge, this is the first report in Italy. The occurrence of this disease may be attributed to excessive irrigation practices. References: (1) S. O. Cacciola et al. EPPO Bull. 20:47, 1990. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) Y. Nagai et al. Phytopathology 68:684, 1978. (4) B. W. Schwingle et al. Plant Dis. 91:97, 2007.


Increasing fatal AA amyloidosis in hunting falcons and how to identify the risk: a report from the United Arab Emirates.
In hunting falcons, a fatal syndrome of wasting, weight loss, green mutes and, finally, sudden death of emaciated birds has been observed in the United Arab Emirates (UAE). Histological examination using Congo red has revealed amyloid in most organs, in particular in the liver, spleen, kidney, and adrenal glands. Moreover, a retrospective study revealed amyloidosis in 100 cases among a total of 623 necropsied falcons between August 1995 and March 2004 in Dubai/UAE (16%; varying from 8 to 30% in different raptor bird species). The amyloid was immunohistochemically classified as amyloid A (AA), which was confirmed by Western blot analysis and N-terminal amino acid sequence analysis, suggesting it to be secondary to a chronic inflammatory process. Retrospective analysis has indicated a significantly increased prevalence of bumble foot and visceral gout among falcons with amyloidosis. In addition, a significant increase of amyloidosis from 5.6% of necropsied falcons with amyloidosis in 1995 to 40.0% in 2004 has been noticed. Finally, a semi-quantitative serum test for falcon serum amyloid A (f-SAA) has been developed. Among 38 falcons with fatal AA amyloidosis, f-SAA was increased pathologically in 36, whereas f-SAA was elevated in only one of 15 apparently disease-free falcons (p < 0.001). This significant result indicates that a normal f-SAA will indicate a minimal or even absent risk of succumbing to AA amyloidosis.


Minichromosome maintenance protein expression according to the grade of atypism in actinic keratosis.
The minichromosome maintenance (MCM) family is a group of proteins that are key initiation factors for DNA replication and are expressed only in cycling cells. Recent studies on various cancerous conditions have shown that MCM proteins are better markers for malignant cells compared to other proliferative markers. It has been also proven that MCM proteins are independent prognostic factors. The aim of this study was to characterize the pattern and frequency of MCM 2 protein expression in actinic keratosis (AK) and determine whether the expression is correlated with the degree of histological atypism. Biopsy samples of 34 patients who had been diagnosed as AK were used in this study. Samples were divided into three groups (grade I, grade II, and grade III) according to the degree of atypism. Immunohistochemical staining for MCM 2 protein, Ki-67, and proliferating cell nuclear antigen was performed, and the number of positively staining cells per unit area (10⁻⁴ μm²) was calculated for evaluation of immunoreactivity. MCM 2 protein was expressed in atypical keratinocytes in AK. Mean numbers of immunoreactive cells positive for MCM 2 were 165.1 in grade I, 304.5 in grade II, and 513.3 in grade III. Moreover, the correlation between the immunoreactivity for MCM 2 protein and AK grade was significantly more positive than that for other markers. Thus, we suggest that MCM 2 protein is a reliable marker for diagnosing and grading AK and further could be hypothesized as an important prognostic factor.


The Unreasonable Effectiveness of Reaction Diffusion in Vertebrate Skin Color Patterning.
In 1952, Alan Turing published the reaction-diffusion (RD) mathematical framework, laying the foundations of morphogenesis as a self-organized process emerging from physicochemical first principles. Regrettably, this approach has been widely doubted in the field of developmental biology. First, we summarize Turing's line of thoughts to alleviate the misconception that RD is an artificial mathematical construct. Second, we discuss why phenomenological RD models are particularly effective for understanding skin color patterning at the meso/macroscopic scales, without the need to parameterize the profusion of variables at lower scales. More specifically, we discuss how RD models (a) recapitulate the diversity of actual skin patterns, (b) capture the underlying dynamics of cellular interactions, (c) interact with tissue size and shape, (d) can lead to ordered sequential patterning, (e) generate cellular automaton dynamics in lizards and snakes, (f) predict actual patterns beyond their statistical features, and (g) are robust to model variations. Third, we discuss the utility of linear stability analysis and perform numerical simulations to demonstrate how deterministic RD emerges from the underlying chaotic microscopic agents.


Predictors and outcomes of joint trajectories of callous-unemotional traits and conduct problems in childhood.
Callous-unemotional (CU) traits are associated with antisocial and delinquent behaviors in children and represent a potential risk factor for adult psychopathy. However, there is a paucity of longitudinal research that explores the development of these traits, their longitudinal association with conduct problems (CP), and their psychosocial predictors and outcomes. Using a large sample of children followed longitudinally from the Twins Early Development Study (N=9,578), we described the joint developmental trajectories of CU traits and CP during childhood (between ages 7 and 12) and examined the child- and family-level predictors (4 years old) and concomitant outcomes (12 years old) associated with the trajectories. The developmental trajectories were characterized with teachers' ratings of CU traits and CP from ages 7 to 12. Using general growth mixture modeling, we identified four trajectories of CU traits (stable high, increasing, decreasing, and stable low) and two trajectories of CP (high and low). Compared with the children who followed a low trajectory of CU traits and CP, those who followed a high trajectory of CU traits and CP had more negative child- and family-level predictors at 4 years (including CP, hyperactivity, negative parental discipline, and chaos in the home). Children with high or increasing levels of CU traits and concomitant high levels of CP presented the most negative outcomes at 12 years (including hyperactivity, peer problems, emotional problems, and negative parental feelings). Children with high CU traits and concomitant high levels of CP in childhood should be prioritized for targeted intervention.


Quantitative Profiling Identifies Potential Regulatory Proteins Involved in Development from Dauer Stage to L4 Stage in Caenorhabditis elegans.
When Caenorhabditis elegans encounters unfavorable growth conditions, it enters the dauer stage, an alternative L3 developmental period. A dauer larva resumes larval development to the normal L4 stage by uncharacterized postdauer reprogramming (PDR) when growth conditions become more favorable. During this transition period, certain heterochronic genes involved in controlling the proper sequence of developmental events are known to act, with their mutations suppressing the Muv (multivulva) phenotype in C. elegans. To identify the specific proteins in which the Muv phenotype is highly suppressed, quantitative proteomic analysis with iTRAQ labeling of samples obtained from worms at L1 + 30 h (for continuous development [CD]) and dauer recovery +3 h (for postdauer development [PD]) was carried out to detect changes in protein abundance in the CD and PD states of both N2 and lin-28(n719). Of the 1661 unique proteins identified with a < 1% false discovery rate at the peptide level, we selected 58 proteins exhibiting ≥2-fold up-regulation or ≥2-fold down-regulation in the PD state and analyzed the Gene Ontology terms. RNAi assays against 15 selected up-regulated genes showed that seven genes were predicted to be involved in higher Muv phenotype (p < 0.05) in lin-28(n791), which is not seen in N2. Specifically, two genes, K08H10.1 and W05H9.1, displayed not only the highest rate (%) of Muv phenotype in the RNAi assay but also the dauer-specific mRNA expression, indicating that these genes may be required for PDR, leading to the very early onset of dauer recovery. Thus, our proteomic approach identifies and quantitates the regulatory proteins potentially involved in PDR in C. elegans, which safeguards the overall lifecycle in response to environmental changes.


Retroviruses.
First brought to scientific attention as infectious cancer-causing agents nearly 80 years ago, retroviruses are popular in contemporary biology for many reasons. (i) The virus life cycle includes several events--in particular, reverse transcription of the viral RNA genome into DNA, orderly integration of viral DNA into host chromosomes, and utilization of host mechanisms for gene expression in response to viral signals--which are broadly informative about eukaryotic cells and viruses. (ii) Retroviral oncogenesis usually depends on transduction or insertional activation of cellular genes, and isolation of those genes has provided the scientific community with many of the molecular components now implicated in the control of normal growth and in human cancer. (iii) Retroviruses include many important veterinary pathogens and two recently discovered human pathogens, the causative agents of the acquired immunodeficiency syndrome (AIDS) and adult T cell leukemia/lymphoma. (iv) Retroviruses are genetic vectors in nature and can be modified to serve as genetic vectors for both experimental and therapeutic purposes. (v) Insertion of retroviral DNA into host chromosomes can be used to mark cell lineages and to make developmental mutants. Progress in these and other areas of retrovirus-related biology has been enormous during the past two decades, but many practical and theoretical problems remain to be solved.


Arachidonic acid stimulates phosphoinositide hydrolysis and human placental lactogen release in an enriched fraction of placental cells.
Previous investigations in this laboratory have indicated that arachidonic acid stimulates a rapid, dose-dependent, and reversible increase in human placental lactogen (hPL) release which is not dependent on cyclooxygenase or lipoxygenase metabolism. To investigate further the mechanism by which arachidonic acid stimulates the release of hPL, the effects of arachidonic acid on phosphoinositide hydrolysis were examined in an enriched cell culture population of term human syncytiotrophoblast. Phosphoinositide hydrolysis was assayed by three methods: the release of 3H from perfused cells prelabeled with [3H]myoinositol, the measurement of inositol phosphate accumulation, and the distribution of radioactivity in phospholipids separated by two-dimensional thin layer chromatography after exposure of 32P-labeled placental cells to arachidonic acid. Arachidonic acid stimulated a concentration-dependent, rapid, and reversible increase in the release of both [3H]myoinositol and hPL from perfused placental cells. This effect was not inhibited by prior incubation of cells with indomethacin (20 microM). In contrast, palmitic acid and oleic acid stimulated phosphoinositide hydrolysis only at a high concentration (100 microM). Arachidonic acid also stimulated the rapid appearance of inositol monophosphate in placental cells. The effect of arachidonic acid was specific for hydrolysis of phosphoinositides and phosphatidylserine and did not involve other phospholipids. Since phosphoinositide hydrolysis is associated with hormone release in a variety of secretory systems, these results suggest that the stimulation of hPL release by arachidonic acid may be mediated, at least in part, by the activation of phospholipase C.


The association between naturally acquired IgG subclass specific antibodies to the PfRH5 invasion complex and protection from Plasmodium falciparum malaria.
Understanding the targets and mechanisms of human immunity to malaria is important for advancing the development of highly efficacious vaccines and serological tools for malaria surveillance. The PfRH5 and PfRipr proteins form a complex on the surface of P. falciparum merozoites that is essential for invasion of erythrocytes and are vaccine candidates. We determined IgG subclass responses to these proteins among malaria-exposed individuals in Papua New Guinea and their association with protection from malaria in a longitudinal cohort of children. Cytophilic subclasses, IgG1 and IgG3, were predominant with limited IgG2 and IgG4, and IgG subclass-specific responses were higher in older children and those with active infection. High IgG3 to PfRH5 and PfRipr were significantly and strongly associated with reduced risk of malaria after adjusting for potential confounding factors, whereas associations for IgG1 responses were generally weaker and not statistically significant. Results further indicated that malaria exposure leads to the co-acquisition of IgG1 and IgG3 to PfRH5 and PfRipr, as well as to other PfRH invasion ligands, PfRH2 and PfRH4. These findings suggest that IgG3 responses to PfRH5 and PfRipr may play a significant role in mediating naturally-acquired immunity and support their potential as vaccine candidates and their use as antibody biomarkers of immunity.


Syntenin and syndecan in the biogenesis of exosomes.
Cells communicate with their environment in various ways, including by secreting vesicles. Secreted vesicles are loaded with proteins, lipids and RNAs that compose 'a signature' of the cell of origin and potentially can reprogram recipient cells. Secreted vesicles recently gained in interest for medicine. They represent potential sources of biomarkers that can be collected from body fluids and, by disseminating pathogenic proteins, might also participate in systemic diseases like cancer, atherosclerosis and neurodegeneration. The mechanisms controlling the biogenesis and the uptake of secreted vesicles are poorly understood. Some of these vesicles originate from endosomes and are called 'exosomes'. In this review, we recapitulate recent insight on the role of the syndecan (SDC) heparan sulphate proteoglycans, the small intracellular adaptor syntenin and associated regulators in the biogenesis and loading of exosomes with cargo. SDC-syntenin-associated regulators include the endosomal sorting complex required for transport accessory component ALG-2-interacting protein X, the small GTPase adenosine 5'-diphosphate-ribosylation factor 6, the lipid-modifying enzyme phospholipase D2 and the endoglycosidase heparanase. All these molecules appear to support the budding of SDC-syntenin and associated cargo into the lumen of endosomes. This highlights a major mechanism for the formation of intraluminal vesicles that will be released as exosomes.


Revisiting Clear Floor Area Requirements for Wheeled Mobility Device Users in Public Transportation.
Current accessibility standards in the U.S. prescribe minimum dimensions for 'clear floor area' to accommodate wheeled mobility device (WhMD) users on transportation vehicles. Prior research on the anthropometry of WhMD users (n = 500) indicates that these dimensions are too small to accommodate the size of many occupied wheeled mobility devices, especially power chairs and scooters. This paper describes a development project designed to update the evidence for these technical criteria and communicate them to the vehicle designers and accessibility standards developers in a manner that would facilitate making good decisions. An interactive web-based design tool was developed for determining the dimensions of clear floor area to achieve a user-specified level of physical accommodation based on occupied device length and width measurements taken on 500 WhMD users. The web-based design tool is now available to practitioners who seek to accommodate a wider range of WhMD users than the minimum standards required by regulations. The design tool is also intended as a visual evidence base for regulatory activity and universal design practice with higher ambitions. The advent of driverless automated vehicles will increase the importance of accessibility and usability to accommodate the diversity of riders with disabilities. Clear floor space to enable independent ingress, interior circulation and egress among WhMD users will be a foremost concern. The transportation industry, standards developers, disability advocates, mobility device manufacturers and prescribers need to understand the limitations of current accessibility standards and work to address these limitations through updated vehicle design standards and policies.


NSAID use and the risk of Parkinson's disease: systematic review and meta-analysis of observational studies.
Several studies have suggested that NSAID use may modify the risk of developing Parkinson's disease (PD). Our aim was to conduct a meta-analysis of observational studies evaluating NSAID use and the risk of PD. We systematically searched MEDLINE (1966-November 2008), EMBASE (1980-November 2008) and other databases. Data from 11 studies were included in the meta-analysis. We used the random effects model to calculate risk ratios (relative risks) and their corresponding 95% confidence intervals (CIs). The pooled risk ratio of PD with NSAID use was 0.95 (95% CI 0.80, 1.12). The pooled risk ratio of PD with high-dose or long-duration NSAID use was 0.91 (95% CI 0.78, 1.05). The pooled risk ratio of PD for aspirin (acetylsalicylic acid) users was 1.08 (95% CI 0.93, 1.26). The pooled risk ratio of PD among ibuprofen users was 0.76 (95% CI 0.65, 0.89). The pooled risk ratio of PD in men using NSAIDs was 0.79 (95% CI 0.69, 0.92), and in women using NSAIDs, it was 0.72 (95% CI 0.45, 1.15). NSAIDs as a class do not seem to modify the risk of PD. However, ibuprofen may have a slight protective effect in lowering the risk of PD. Although the risk ratios of PD in male and female NSAID users were similar, the 95% CI for men was suggestive of a slight risk reduction.