Biophysical properties of the human finger for touch comprehension: influences of ageing and gender.
The human finger plays an extremely important role in tactile perception, but little is known about how age and gender affect its biophysical properties and their role in tactile perception. We combined studies on contact characteristics, mechanical properties and surface topography to understand age and gender effects on the human finger. The values obtained regarding contact characteristics (i.e. adhesive force) were significantly higher for women than for men. As for mechanical properties (i.e. Young's modulus E), a significant and positive correlation with age was observed and found to be higher for women. A positive correlation was observed between age and the arithmetic mean of surface roughness for men. However, an inverse age effect was highlighted for women. The age and gender effects obtained have never been reported previously in the literature. These results open new perspectives for understanding the weakening of tactile perception across ages and how it differs between men and women.


Identification of a Novel Hypovirulence-Inducing Hypovirus From Alternaria alternata.
Mycoviruses are wide spread throughout almost all groups of fungi but only a small number of mycoviruses can attenuate the growth and virulence of their fungal hosts. Alternaria alternata is an ascomycete fungus that causes leaf spot diseases on various crop plants. In this study, we identified a novel ssRNA mycovirus infecting an A. alternata f. sp. mali strain isolated from an apple orchard in China. Sequence analyses revealed that this virus is related to hypoviruses, in particular to Wuhan insect virus 14, an unclassified hypovirus identified from insect meta-transcriptomics, as well as other hypoviruses belonging to the genus Hypovirus, and therefore this virus is designed as Alternaria alternata hypovirus 1 (AaHV1). The genome of AaHV1 contains a single large open-reading frame encoding a putative polyprotein (∼479 kDa) with a cysteine proteinase-like and replication-associated domains. Curing AaHV1 from the fungal host strain indicated that the virus is responsible for the slow growth and reduced virulence of the host. AaHV1 defective RNA (D-RNA) with internal deletions emerging during fungal subcultures but the presence of D-RNA does not affect AaHV1 accumulation and pathogenicities. Moreover, AaHV1 could replicate and confer hypovirulence in Botryosphaeria dothidea, a fungal pathogen of apple white rot disease. This finding could facilitate better understanding of A. alternata pathogenicity and is relevant for development of biocontrol methods of fungal diseases.


The fetal liver as cell source for the regenerative medicine of liver and pancreas.
Patients affected by liver diseases and diabetes mellitus are in need for sources of new cells to enable a better transition into clinic programs of cell therapy and regenerative medicine. In this setting, fetal liver is becoming the most promising and available source of cells. Fetal liver displays unique characteristics given the possibility to isolate cell populations with a wide spectrum of endodermal differentiation and, the co-existence of endodermal and mesenchymal-derived cells. Thus, the fetal liver is a unique and highly available cell source contemporarily candidate for the regenerative medicine of both liver and pancreas. The purpose of this review is to revise the recent literature on the different stem cells populations isolable from fetal liver and candidate to cell therapy of liver diseases and diabetes and to discuss advantages and limitation with respect to other cell sources.


Growth rates of intracranial aneurysms: exploring constancy.
The annual rate of rupture of intracranial aneurysms is often assumed to be constant, but it is unknown whether this assumption is true. Recent case reports have suggested that aneurysms grow fast in a short period of time. The authors of the present report investigated the plausibility of a constant growth rate for intracranial aneurysms. Assuming a constant aneurysm growth rate within an individual and varying rates between individuals, a hypothetical cohort was simulated. Individuals with high growth rates will display aneurysm formation and rupture at a young age; such persons disappear early from the hypothetical cohort. As a result the mean lesion growth rate varies over time. In hypothetical cohorts with different initial mean growth rates, the authors calculated age-specific incidence rates (per 100,000 person-years) of subarachnoid hemorrhage and compared these rates with population-based data on the incidence of subarachnoid hemorrhage (per 100,000 person-years). A hypothetical cohort with a mean initial growth rate of 0.18 mm/year reproduced most closely the incidence rates observed in the population. However, even for this most plausible hypothetical cohort, age-specific incidence rates in the model differed substantially and statistically significantly from those observed in the population. Based on the results of this study, it is unlikely that intracranial aneurysms in general grow at a constant time-independent rate. The authors hypothesized that the actual growth process is irregular and discontinuous, which results in periods with and without aneurysm growth and with high and low risks of rupture.


Preconditioning for multiplexed imaging with spatially coded PSFs.
We propose a preconditioning method to improve the convergence of iterative reconstruction algorithms in multiplexed imaging based on convolution-based compressive sensing with spatially coded point spread functions (PSFs). The system matrix is converted to improve the condition number with a preconditioner matrix. The preconditioner matrix is calculated by Tikhonov regularization in the frequency domain. The method was demonstrated with simulations and an experiment involving a range detection system with a grating based on the multiplexed imaging framework. The results of the demonstrations showed improved reconstruction fidelity by using the proposed preconditioning method.


Eukaryotic core promoters and the functional basis of transcription initiation.
RNA polymerase II (Pol II) core promoters are specialized DNA sequences at transcription start sites of protein-coding and non-coding genes that support the assembly of the transcription machinery and transcription initiation. They enable the highly regulated transcription of genes by selectively integrating regulatory cues from distal enhancers and their associated regulatory proteins. In this Review, we discuss the defining properties of gene core promoters, including their sequence features, chromatin architecture and transcription initiation patterns. We provide an overview of molecular mechanisms underlying the function and regulation of core promoters and their emerging functional diversity, which defines distinct transcription programmes. On the basis of the established properties of gene core promoters, we discuss transcription start sites within enhancers and integrate recent results obtained from dedicated functional assays to propose a functional model of transcription initiation. This model can explain the nature and function of transcription initiation at gene starts and at enhancers and can explain the different roles of core promoters, of Pol II and its associated factors and of the activating cues provided by enhancers and the transcription factors and cofactors they recruit.


The endophytic fungus Piriformospora indica reprograms barley to salt-stress tolerance, disease resistance, and higher yield.
Disease resistance strategies are powerful approaches to sustainable agriculture because they reduce chemical input into the environment. Recently, Piriformospora indica, a plant-root-colonizing basidiomycete fungus, has been discovered in the Indian Thar desert and was shown to provide strong growth-promoting activity during its symbiosis with a broad spectrum of plants. Here, we report on the potential of P. indica to induce resistance to fungal diseases and tolerance to salt stress in the monocotyledonous plant barley. The beneficial effect on the defense status is detected in distal leaves, demonstrating a systemic induction of resistance by a root-endophytic fungus. The systemically altered "defense readiness" is associated with an elevated antioxidative capacity due to an activation of the glutathione-ascorbate cycle and results in an overall increase in grain yield. Because P. indica can be easily propagated in the absence of a host plant, we conclude that the fungus could be exploited to increase disease resistance and yield in crop plants.


Prediction errors to emotional expressions: the roles of the amygdala in social referencing.
Social referencing paradigms in humans and observational learning paradigms in animals suggest that emotional expressions are important for communicating valence. It has been proposed that these expressions initiate stimulus-reinforcement learning. Relatively little is known about the role of emotional expressions in reinforcement learning, particularly in the context of social referencing. In this study, we examined object valence learning in the context of a social referencing paradigm. Participants viewed objects and faces that turned toward the objects and displayed a fearful, happy or neutral reaction to them, while judging the gender of these faces. Notably, amygdala activation was larger when the expressions following an object were less expected. Moreover, when asked, participants were both more likely to want to approach, and showed stronger amygdala responses to, objects associated with happy relative to objects associated with fearful expressions. This suggests that the amygdala plays two roles in social referencing: (i) initiating learning regarding the valence of an object as a function of prediction errors to expressions displayed toward this object and (ii) orchestrating an emotional response to the object when value judgments are being made regarding this object.


Transcranial magnetic simulation in the treatment of migraine.
Transcranial magnetic stimulation (TMS) is a diagnostic and therapeutic modality that is being developed as both an acute and preventive treatment for migraine. TMS delivers a fluctuating magnetic field from the scalp surface to induce current in the subjacent cortex. Magnetic pulses are delivered one at a time in single-pulse TMS (sTMS) or as a train of pulses in repetitive TMS (rTMS). For most of its 30-year history, TMS has been delivered in clinical and research settings using large tabletop devices. Based on the theory that sTMS may disrupt cortical spreading depression, sTMS has been studied and shown to be effective as an acute treatment for migraine with aura. Subsequent work in animal models confirms that sTMS disrupts cortical spreading depression. To make outpatient self-treatment possible, a portable device has been developed for acute treatment of migraine with aura. Based on the theory that rTMS alters brain excitability and neurotransmitter activity, rTMS has been studied as a preventive migraine treatment. A small body of evidence suggests that rTMS may have a role, but further studies are needed. In this review, we summarize the data on TMS as a treatment of migraine, and we suggest directions for future research.


Effects of bigeminies and paired-pulse stimulation on oxygen consumption in dog left ventricle.
In 10 excised, cross-circulated, isovolumically beating dog left ventricles, we examined the effects of bigeminy, including paired-pulse stimulation, on cardiac oxygen consumption (VO2) and evaluated whether the VO2 versus pressure-volume area (PVA) relation obtained from regularly beating hearts in our previous studies could account for the changes in VO2 during bigeminies with various coupling intervals. The extrasystolic interval (ESI) was decreased in four steps from 300 msec (regular rhythm) to paired-pulse stimulation (less than or equal to 210 msec). The sum of ESI and the postextrasystolic interval was always held constant (600 msec); therefore, the number of excitations was 200/min at any ESI. Both VO2 and PVA were measured in four runs: a small volume run at a ventricular volume of 13 ml, a large volume of 12 ml, a calcium run in which contractility was enhanced by CaCl2 at a small volume of 12 ml, and a verapamil run in which contractility was depressed by verapamil at a large volume of 20 ml. In any run, both VO2 and PVA were unchanged at long ESIs (greater than or equal to 250 msec) from each control value at regular rhythm, but VO2 markedly increased at short ESIs (less than or equal to 230 msec). VO2s during bigeminies with various ESIs were comparable with the theoretical VO2 values that were calculated by the VO2-PVA relation at regular rhythm. However, the theoretical VO2 values underestimated the measured VO2 values at the shortest ESIs under a high volume loading and in a high contractile state. This indicates that the PVA concept can be expanded to various arrhythmias unless ESI becomes very short.


Total lymphoid irradiation: a novel and successful therapy for resistant cardiac allograft rejection.
Total lymphoid irradiation (TLI) is a novel type of adjuvant immunosuppression for patients who undergo cardiac transplantation and have refractory allograft rejection during standard immunosuppressive therapy. TLI consists of 6 to 10 fractions of 80 cGy (1 cGy = 1 rad) of irradiation to lymphatic tissues with use of the standard mantle and inverted Y fields. We have used TLI in six patients with biopsy-proven rejection that was refractory to standard treatments, including cyclosporine, azathioprine, antilymphocyte antibodies, and corticosteroids. In five patients, recalcitrant rejection was resolved after completion of TLI, and resolution persisted during long-term follow-up (17 to 30 months; mean, 22.2 months). In each patient, a substantial increase in the CD8 (suppressor T-lymphocyte) subset and elimination of B lymphocytes were demonstrated, findings that also persisted. Side effects were mild and primarily limited to transient leukopenia. In four patients, a readily treated cytomegalovirus reactivation was noted during TLI; thus, a causal relationship was suggested. In recipients of cardiac allografts who have refractory rejection, TLI provides long-lasting amelioration of the rejection profile. This result may be attributable to a relative enhancement of the suppressor T-cell subset and elimination of the B-lymphocyte line. Side effects are minimal, but monitoring for cytomegalovirus activation or reactivation is recommended.


Analysis of the evolutionary relationships of HIV-1 and SIVcpz sequences using bayesian inference: implications for the origin of HIV-1.
The most plausible origin of HIV-1 group M is an SIV lineage currently represented by SIVcpz isolated from the chimpanzee subspecies Pan troglodytes troglodytes. The origin of HIV-1 group O is less clear. Putative recombination between any of the HIV-1 and SIVcpz sequences was tested using bootscanning and Bayesian-scanning plots, as well as a new method using a Bayesian multiple change-point (BMCP) model to infer parental sequences and crossing-over points. We found that in the case of highly divergent sequences, such as HIV-1/SIVcpz, Bayesian scanning and BMCP methods are more appropriate than bootscanning analysis to investigate spatial phylogenetic variation, including estimating the boundaries of the regions with discordant evolutionary relationships and the levels of support of the phylogenetic clusters under study. According to the Bayesian scanning plots and BMCP method, there was strong evidence for discordant phylogenetic clustering throughout the genome: (1) HIV-1 group O clustered with SIVcpzANT/TAN in middle pol, and partial vif/env; (2) SIVcpzGab1 clustered with SIVcpzANT/TAN in 3'pol/vif, and middle env; (3) HIV-1 group O grouped with SIVcpzCamUS and SIVcpzGab1 in p17/p24; (4) HIV-1 group M was more closely related to SIVcpzCamUS in 3'gag/pol and in middle pol, whereas in partial gp120 group M clustered with group O. Conditionally independent phylogenetic analysis inferred by maximum likelihood (ML) and Bayesian methods further confirmed these findings. The discordant phylogenetic relationships between the HIV-1/SIVcpz sequences may have been caused by ancient recombination events, but they are also due, at least in part, to altered rates of evolution between parental SIVcpz lineages.


Clinical Experience With 75-mg Dose of Erlotinib for Mutated Metastatic EGFR Non-small Cell Lung Cancer.
Non-small cell lung cancer (NSCLC) remains the leading cause of cancer death worldwide, although important advances in target therapy have been developed in the past few years. Erlotinib is a reversible epidermal growth factor receptor (EGFR) inhibitor, which was approved at its maximum tolerated dose of 150 mg/d determined from the initial phase I study. Studies suggest that the optimal biological dose of erlotinib should be lower and dependent on different variables. We aimed to evaluate the response rates and toxicity with 75 mg/d dose of erlotinib in South American patients. We performed a retrospective review of 18 patients with histologically proven (+) EGFR (+) mutation metastatic NSCLC (mNSCLC) treated with 75 mg/d erlotinib as starting dose. Clinical information, including toxicity grade 1-4, drug discontinuation, clinical evolution and radiological evaluation, and overall survival (OS), was revised. Patients received 75 mg/d of erlotinib as starting dose. Sixteen (89%) patients were treated in first-line treatment and 2 (11%) in second-line treatment. Mean age was 62 years (range 36-89 years), and 50% patients were female. Sixteen percent of the patients had brain metastases at first diagnosis. All patients had mutation positive EGFR, 12 (66%) had Del19 and 6 (34%) exon 21 mutation. Median progression-free survival was 17 months and OS 23 months. The main grade 1-2 toxicities were rash (44%) and diarrhea (22%). No grade 3-4 toxicity and no cases of drug discontinuation were reported. In South American population with mutated mNSCLC, a dose of 75 mg/d of erlotinib was well tolerated. This dose resulted in comparable benefits in progression-free survival and OS when compared to those reported in the literature with the standard dose. More studies are needed to explore the use of adjusted doses of biological agents in different ethnic backgrounds.


Expanding the clinical and mutational spectrum of the Ehlers-Danlos syndrome, dermatosparaxis type.
The Ehlers-Danlos syndrome (EDS), dermatosparaxis type, is a recessively inherited connective tissue disorder caused by deficient activity of ADAMTS-2, an enzyme that cleaves the aminoterminal propeptide domain of types I, II, and III procollagen. Only 10 EDS dermatosparaxis patients have been reported, all presenting a recognizable phenotype with characteristic facial gestalt, extreme skin fragility and laxity, excessive bruising, and sometimes major complications due to visceral and vascular fragility. We report on five new EDS dermatosparaxis patients and provide a comprehensive overview of the current knowledge of the natural history of this condition. We identified three novel homozygous loss-of-function mutations (c.2927_2928delCT, p.(Pro976Argfs*42); c.669_670dupG, p.(Pro224Argfs*24); and c.2751-2A>T) and one compound heterozygous mutation (c.2T>C, p.? and c.884_887delTGAA, p.(Met295Thrfs26*)) in ADAMTS2 in five patients from four unrelated families. Three of these displayed a phenotype that was strikingly milder than that of previously reported patients. This study expands the clinical and molecular spectrum of the dermatosparaxis type of EDS to include a milder phenotypic variant and stresses the importance of good clinical criteria. To address this, we propose an updated set of criteria that accurately captures the multisystemic nature of the dermatosparaxis type of EDS.Genet Med 18 9, 882-891.


Genetic differences between the determinants of lipid profile phenotypes in African and European Americans: the Jackson Heart Study.
Genome-wide association analysis in populations of European descent has recently found more than a hundred genetic variants affecting risk for common disease. An open question, however, is how relevant the variants discovered in Europeans are to other populations. To address this problem for cardiovascular phenotypes, we studied a cohort of 4,464 African Americans from the Jackson Heart Study (JHS), in whom we genotyped both a panel of 12 recently discovered genetic variants known to predict lipid profile levels in Europeans and a panel of up to 1,447 ancestry informative markers allowing us to determine the African ancestry proportion of each individual at each position in the genome. Focusing on lipid profiles -- HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), and triglycerides (TG) -- we identified the lipoprotein lipase (LPL) locus as harboring variants that account for interethnic variation in HDL-C and TG. In particular, we identified a novel common variant within LPL that is strongly associated with TG (p = 2.7 x 10(-6)) and explains nearly 1% of the variability in this phenotype, the most of any variant in African Americans to date. Strikingly, the extensively studied "gain-of-function" S447X mutation at LPL, which has been hypothesized to be the major determinant of the LPL-TG genetic association and is in trials for human gene therapy, has a significantly diminished strength of biological effect when it is found on a background of African rather than European ancestry. These results suggest that there are other, yet undiscovered variants at the locus that are truly causal (and are in linkage disequilibrium with S447X) or that work synergistically with S447X to modulate TG levels. Finally, we find systematically lower effect sizes for the 12 risk variants discovered in European populations on the African local ancestry background in JHS, highlighting the need for caution in the use of genetic variants for risk assessment across different populations.


RE-CODE DCM (REsearch Objectives and Common Data Elements for Degenerative Cervical Myelopathy): A Consensus Process to Improve Research Efficiency in DCM, Through Establishment of a Standardized Dataset for Clinical Research and the Definition of the Research Priorities.
Mixed-method consensus process. Degenerative cervical myelopathy (DCM) is a common and disabling condition that arises when mechanical stress damages the spinal cord as a result of degenerative changes in the surrounding spinal structures. RECODE-DCM (REsearch Objectives and Common Data Elements for Degenerative Cervical Myelopathy) aims to improve efficient use of health care resources within the field of DCM by using a multi-stakeholder partnership to define the DCM research priorities, to develop a minimum dataset for DCM clinical studies, and confirm a definition of DCM. This requires a multi-stakeholder partnership and multiple parallel consensus development processes. It will be conducted via 4 phases, adhering to the guidance set out by the COMET (Core Outcomes in Effectiveness Trials) and JLA (James Lind Alliance) initiatives. Phase 1 will consist of preliminary work to inform online Delphi processes (Phase 2) and a consensus meeting (Phase 3). Following the findings of the consensus meeting, a synthesis of relevant measurement instruments will be compiled and assessed as per the COSMIN (Consensus-based Standards for the Selection of Health Measurement Instruments) criteria, to allow recommendations to be made on how to measure agreed data points. Phase 4 will monitor and promote the use of eventual recommendations. RECODE-DCM sets out to establish for the first time an index term, minimum dataset, and research priorities together. Our aim is to reduce waste of health care resources in the future by using patient priorities to inform the scope of future DCM research activities. The consistent use of a standard dataset in DCM clinical studies, audit, and clinical surveillance will facilitate pooled analysis of future data and, ultimately, a deeper understanding of DCM.


Thymic generation and selection of CD25+CD4+ regulatory T cells: implications of their broad repertoire and high self-reactivity for the maintenance of immunological self-tolerance.
Besides positive and negative selection of T cells, another function of the thymus in maintaining immunological self-tolerance is the production of CD25+CD4+ regulatory T cells capable of preventing autoimmune disease. They acquire the regulatory activity through the thymic selection process, and are released to the periphery as a functionally and phenotypically mature population. Our recent study with transgenic mice in which every class II MHC molecule covalently binds the same single peptide has revealed that a particular self-peptide/MHC ligand in the thymus can positively select a broad repertoire of functionally mature CD25+CD4+ regulatory T cells as well as naive T cells. Interestingly, the regulatory T cells bear higher reactivity than other T cells to the selecting ligand in the thymus even after negative selection by the ligand. This broad repertoire and high self-reactivity of CD25+CD4+ regulatory T cells, together with their high level expression of various accessory molecules, may guarantee their prompt and efficient activation upon encounter with a diverse range of self peptide/MHC complexes in the periphery, ensuring dominant control of self-reactive T cells.


Corticolimbic hyper-response to emotion and glutamatergic function in people with high schizotypy: a multimodal fMRI-MRS study.
Animal models and human neuroimaging studies suggest that altered levels of glutamatergic metabolites within a corticolimbic circuit have a major role in the pathophysiology of schizophrenia. Rodent models propose that prefrontal glutamate dysfunction could lead to amygdala hyper-response to environmental stress and underlie hippocampal overdrive in schizophrenia. Here we determine whether changes in brain glutamate are present in individuals with high schizotypy (HS), which refers to the presence of schizophrenia-like characteristics in healthy individuals, and whether glutamate levels are related to altered corticolimbic response to emotion. Twenty-one healthy HS subjects and 22 healthy subjects with low schizotypy (LS) were selected based on their Oxford and Liverpool Inventory of Feelings and Experiences rating. Glutamate levels were measured in the anterior cingulate cortex (ACC) using proton magnetic resonance spectroscopy, followed by a functional magnetic resonance imaging (fMRI) scan to measure corticolimbic response during emotional processing. fMRI results and fMRI × glutamate interactions were considered significant after voxel-wise P<0.05 family-wise error correction. While viewing emotional pictures, HS individuals showed greater activation than did subjects with LS in the caudate, and marginally in the ACC, hippocampus, medial prefrontal cortex (MPFC) and putamen. Although no between-group differences were found in glutamate concentrations, within the HS group ACC glutamate was negatively correlated with striatal activation (left: z=4.30, P=0.004 and right: z=4.12 P=0.008 caudate; left putamen: z=3.89, P=0.018) and marginally with MPFC (z=3.55, P=0.052) and amygdala (left: z=2.88, P=0.062; right: z=2.79, P=0.079), correlations that were not present in LS subjects. These findings provide, to our knowledge, the first evidence that brain glutamate levels are associated with hyper-responsivity in brain regions thought to be critical in the pathophysiology of psychosis.


Function of tightly bound nucleotides on membrane-bound chloroplast coupling factor.
The kinetic behavior of tightly bound nucleotides on chloroplast coupling factor from spinach was determined under phosphorylating and nonphosphorylating conditions. Chloroplast coupling factor 1 (CF1) was labeled with tightly bound radioactive ADP and/or ATP at two specific sites and reconstituted with thylakoid membranes depleted of CF1 by treatment with NaBr. The initial incorporation and dissociation of ADP from one of the sites requires light but occurs at the same rate under phosphorylating and non-phosphorylating conditions. The initial rate is considerably slower than the rate of ATP synthesis, but nucleotide exchange is very rapid during steady-state ATP synthesis. A direct correspondence between this nucleotide binding site and a site on soluble CF1 that hydrolyzes ATP was demonstrated. A second site binds MgATP very tightly; the MgATP does not dissociate during ATP synthesis nor does its presence alter the rate of ATP synthesis. This is analogous to the behavior found for soluble CF1 during ATP hydrolysis. These results demonstrate that the tight-binding nucleotide sites on soluble CF1 and membrane-bound coupling factor are essentially identical in terms of binding properties and kinetic behavior during ATP hydrolysis and synthesis.


Modeling of beam width in passively Q-switched end-pumped lasers.
An iterative model based on the LambertW function was developed for estimating the fundamental mode parameters of resonators with saturable gain guiding. The process of pulse buildup in passively Qswitched, end-pumped lasers was analyzed. The effective ABCD cavity matrix for consecutive round-trips was calculated, taking into account spatially variable saturated gain in an active medium and absorption bleaching in a saturable absorber. The twofold decrease in beam width, as compared with the fundamental mode of the bare cavity, was demonstrated. The application of such a model for resonators with other nonlinear elements is feasible.


Evidence that the severity of depletion of inorganic phosphate determines the severity of the disturbance of adenine nucleotide metabolism in the liver and renal cortex of the fructose-loaded rat.
To test the hypothesis that in both the liver and renal cortex of the fructose-loaded rat, severity of depletion of inorganic phosphate (P(i)), and not the magnitude of accumulation of fructose-1-phosphate (F-1-P), determines the severity of the dose-dependent reduction of ATP, we intraperitoneally injected fed rats with fructose, 20 and 40 mumol/g, alone, and at the higher load, in combination with (a) sodium phosphate, 20 mumol/g, administered shortly beforehand or subsequently or, (b) adenosine, 2 mumol/g, administered beforehand. The following observations were made: (a) With fructose loading alone, at the higher load, both P(i) and total adenine nucleotides (TAN) were reduced by one third in the renal cortex and (as previously observed) by two thirds in the liver; and at either load, the reduction of ATP (and TAN) and the accumulation of F-1-P were less severe in the renal cortex than in the liver. (b) Prior phosphate loading largely prevented the reductions of ATP and TAN in the renal cortex and significantly attenuated them in the liver, yet doubled the renal cortical accumulation of F-1-P. (c) Adenosine loading substantially attenuated the reductions of ATP, TAN, and P(i) only in the renal cortex. (d) ATP varied directly with P(i) (P < 0.001, r = 0.98) in the domain of control and reduced values of P(i) taken from both liver and renal cortex. (e) As judged from tissue and plasma concentrations of fructose and glucose, and tissue concentrations of fructose-6-phosphate and glucose-6-phosphate, the rate at which the renal cortex and liver converted fructose to glucose was much lower at the higher fructose load. (f) Prior phosphate loading prevented this decrease in rate in the renal cortex and attenuated it in the liver; adenosine loading attenuated it only in the renal cortex. We conclude that in both the renal cortex of the fructose-loaded rat: (a) Depletion of P(i) is critical to the causation of the reductions in both ATP and TAN and, at the higher fructose load, of a decrease in the rate at which ATP is regenerated. (b) The severity of depletion of P(i) determines the severity of these disturbances. (c) By differentially mitigating the depletion of P(i), prior phosphate loading largely prevents these disturbances in the renal cortex, and attenuates them in the liver; and adenosine loading attenuates them only in the renal cortex. The findings provide some basis for the observation that in patients with hereditary fructose intolerance experimentally exposed to fructose, prior loading with sodium phosphate substantially attenuates the renal but not hepatic dysfunction.


Quantifying the trigger level of the vacuum surveillance system of the Gamma-Knife eXtend™ positioning system and evaluating the potential impact on dose delivery.
This work evaluates the precision and characteristics of the trigger level of the vacuum surveillance system of eXtend™ on Gamma Knife Perfexion and the effect of the potential displacement on the dose distribution. A total of 20 individually moulded mouthpieces based on human dental models were used to measure translational shift and rotation until the vacuum surveillance of eXtend interrupted the irradiation. The positional accuracy of the movement was 0.01 mm using a computerized numerically controlled positioning system. Rotation was introduced by peripheral pressure in superior or inferior direction on the mould and was measured with a digital inclinometer. In 10 patients with a large brain metastasis the effect of a potential displacement of the centre of the target was recalculated. Two out of the 10 targets were located near the optic nerve and chiasm. In addition, the potential displacement of the chiasm and the optic nerve due to rotation based on their distance to the centre of rotation (COR, the mouthpiece) was evaluated. For the recalculation of the delivered dose after a displacement, GammaPlan Software® was used. Dose distribution with displacement between ±0.1 mm and ±2.0 mm was simulated for ten targets with volumes between 7.7 cm3 and 19.3 cm3. Shifts were applied to the X (lateral), Y (ventro-dorsal) and Z axis (superior-inferior). Dwell time was kept constant within <0.5% total time and 1% individual dwell time. Evaluated parameters were "minimum dose" (D(99%)), "coverage" and "Paddick Conformity Index" (PCI). The vacuum between mouthpiece and dental model was broken by a mean translation of the mouthpiece of 0.15 mm (SD ±0.05 mm, range 0.05-0.29 mm) or a mean rotation of 0.33º (SD ±0.15º, range 0.05-1.0º). Rotation caused by pressure in the inferior direction was higher with 0.42º (SD ±0.16º) compared to rotation due to pressure in the superior direction with 0.21º (SD ±0.09º). The mean rotation would produce a mean displacement of 0.64 mm for the target, 0.39 mm for the chiasm and 0.33 mm for the optic nerve, for the group of patients studied. The biggest effect of positional deviation was seen in the minimum dose to the target, with 2.5% reduction for a 0.5 mm and 5% for a 1.0 mm shift in the Z-direction respectively. The trigger level is sensitive and reproducible. A sudden movement by the patient, such as an attempt to get out or correct an uncomfortable position would interrupt irradiation immediately. The dominant effect for a displacement is due to rotation. The resulting displacement depends on the distance between target and COR (magnitude) and the location (direction). Therefore, lesions near chiasm or brainstem, which are closer to the COR, are less affected from a rotation than peripheral targets.


The fractional contributions of elementary modes to the metabolism of Escherichia coli and their estimation from reaction entropies.
The metabolism of a cell can be viewed as a weighted sum of elementary modes. Due to the multiplicity of modes the identification of the individual weights represents a non-trivial problem. To enable the determination of weighting factors we have identified and implemented two gene deletions in combination with defined growth conditions that limit the metabolism from 4374 original elementary modes to 24 elementary modes for a non-PHB synthesizing control and 40 modes for a PHB synthesizing strain. These remaining modes can be further grouped into five families that have the same overall stoichiometry. Thus, the complexity of the problem is significantly reduced, and weighting factors for each family of modes could be determined from the measurement of accumulation rates of metabolites. Moreover, it is shown that individual weights are inversely correlated with the entropy generated by the operation of the used pathways defined in elementary modes. This suggests that evolution developed cellular regulatory patterns that permit diversity of pathways while favoring efficient pathways with low entropy generation. Furthermore, such correlation provides a rational way of estimating metabolic fluxes based on the thermodynamic properties of elementary modes. This is demonstrated with an example in which experimentally determined, intracellular fluxes are shown to be highly correlated with fluxes computed based on elementary modes and reaction entropies. The analysis suggests that the set of elementary modes can be interpreted analogous to a metabolic ensemble of quantum states of a macroscopic system.


The acute satellite cell response and skeletal muscle hypertrophy following resistance training.
The extent of skeletal muscle hypertrophy in response to resistance training is highly variable in humans. The main objective of this study was to explain the nature of this variability. More specifically, we focused on the myogenic stem cell population, the satellite cell (SC) as a potential mediator of hypertrophy. Twenty-three males (aged 18-35 yrs) participated in 16 wk of progressive, whole body resistance training, resulting in changes of 7.9±1.6% (range of -1.9-24.7%) and 21.0±4.0% (range of -7.0 to 51.7%) in quadriceps volume and myofibre cross-sectional area (CSA), respectively. The SC response to a single bout of resistance exercise (80% 1RM), analyzed via immunofluorescent staining resulted in an expansion of type II fibre associated SC 72 h following exercise (pre: 11.3±0.9; 72 h: 14.8±1.4 SC/type II fibre; p<0.05). Training resulted in an expansion of the SC pool associated with type I (pre: 10.7±1.1; post: 12.1±1.2 SC/type I fibre; p<0.05) and type II fibres (pre: 11.3±0.9; post: 13.0±1.2 SC/type II fibre; p<0.05). Analysis of individual SC responses revealed a correlation between the relative change in type I associated SC 24 to 72 hours following an acute bout of resistance exercise and the percentage increase in quadriceps lean tissue mass assessed by MRI (r2 = 0.566, p = 0.012) and the relative change in type II associated SC following 16 weeks of resistance training and the percentage increase in quadriceps lean tissue mass assessed by MRI (r2 = 0.493, p = 0.027). Our results suggest that the SC response to resistance exercise is related to the extent of muscular hypertrophy induced by training.


The prevalence of emotional disorder in children.
This report presents data from the Ontario Child Health Study on the prevalence of individual emotional symptoms, the variation of prevalence rates of emotional disorders as a function of demographic factors, and the rates of service utilization for children with these disorders. To survey Ontario children from 4 to 16 years of age, a stratified random sample of 3,294 children was drawn from all household dwellings listed in the 1981 Census. The prevalence rates for emotional disorders in male and female children aged 4 to 11 years were 10.1% and 10.7%, respectively. For adolescents aged 12 to 16 years, the prevalence rate rose to 13.7% for females and fell to 5.0% for males. Children with emotional disorders were frequently found to meet criteria for more than one disorder. Only one in five children with emotional disorders had received mental health or social services in the 6 months prior to the survey.


Numerical characterization of whispering-gallery mode optical microcavities.
We characterize planar microcavities in whispering-gallery mode optical resonances. The microcavity consists of a waveguide and a microdisk, and a nanoscale gap separates the waveguide and the microdisk. The devices can be fabricated on Si-based thin films by using conventional microelectronics techniques. To characterize these types of cavity, we study a broad range of resonator configuration parameters including the size of the microdisk, the width of the gap, and the waveguide dimensions. The finite-element method is used for solving Maxwell's equations. The electric fields and the energy density distributions are obtained and compared between the on-resonance and off-resonance situations. A brilliant ring with a strong electric field and a high-energy density is found inside the periphery of the microdisk under first-order resonance. While under second-order resonance, there are two bright rings, and the light intensity in the inner ring is stronger than that in the outer ring. The resonant frequencies and their free spectral ranges are predominantly determined by the size of the microdisk. The gap effect on the resonant frequencies is observable, although it is minor. The gap strongly affects the full width at half-maximum (FWHM), finesse, and quality factor of the resonances. With an increase in the gap width from 100 to 300 nm, both the Q value and finesse increase substantially, while the FWHM decreases. The waveguide width has a visible influence on the Q value, FWHM, and finesse as well.


MoS2 heterostructure with tunable phase stability: strain induced interlayer covalent bond formation.
The structural phase transition in MoS2 promises applications in novel nanoelectronic devices. Elastic strain engineering can not only serve as a potential route for phase transition engineering, but also reveal potential ferroelastic behavior of MoS2 nanostructures. However, the elastic strain required for phase transition in monolayer MoS2 is far beyond its elastic limit, thus inhibiting the experimental realization. In this study, employing density functional theory calculations, we uncover that by forming heterostructure with buckled 2D materials, such as silicene, germanene and stanene, the critical phase transition strain required in monolayer MoS2 can be drastically reduced. Particularly when MoS2 forms sandwiched structures with silicene or stanene, the uniaxial and biaxial critical strain can be reduced to ∼0.06 and ∼0.03, respectively, which is well below the experimental elastic limit. This theoretical study not only proposes an experimental achievable strategy for flexible phase transition design in MoS2 nanostructure, but also identifies those MoS2 heterostructures as 2D candidates for potential shape memory devices and pseudoelasticity applications.


Penetration, distribution and brain toxicity of titanium nanoparticles in rodents' body: a review.
Titanium dioxide (TiO2) has been vastly used commercially, especially as white pigment in paints, colorants, plastics, coatings, cosmetics. Certain industrial uses TiO2 in diameter <100 nm. There are three common exposure routes for TiO2: (i) inhalation exposure, (ii) exposure via gastrointestinal tract, (iii) dermal exposure. Inhalation and gastrointestinal exposure appear to be the most probable ways of exposure, although nanoparticle (NP) penetration is limited. However, the penetration rate may increase substantially when the tissue is impaired. When TiO2 NPs migrate into the circulatory system, they can be distributed into all tissues including brain. In brain, TiO2 lead to oxidative stress mediated by the microglia phagocytic cells which respond to TiO2 NPs by the production and release of superoxide radicals that convert to multiple reactive oxygen species (ROS). The ROS production may also cause the damage of blood-brain barrier which then becomes more permeable for NPs. Moreover, several studies have showed neuron degradation and the impairment of spatial recognition memory and learning abilities in laboratory rodent exposed to TiO2 NPs.


GATA1-mediated megakaryocyte differentiation and growth control can be uncoupled and mapped to different domains in GATA1.
The DNA-binding hemopoietic zinc finger transcription factor GATA1 promotes terminal megakaryocyte differentiation and restrains abnormal immature megakaryocyte expansion. How GATA1 coordinates these fundamental processes is unclear. Previous studies of synthetic and naturally occurring mutant GATA1 molecules demonstrate that DNA-binding and interaction with the essential GATA1 cofactor FOG-1 (via the N-terminal finger) are required for gene expression in terminally differentiating megakaryocytes and for platelet production. Moreover, acquired mutations deleting the N-terminal 84 amino acids are specifically detected in megakaryocytic leukemia in human Down syndrome patients. In this study, we have systematically dissected GATA1 domains required for platelet release and control of megakaryocyte growth by ectopically expressing modified GATA1 molecules in primary GATA1-deficient fetal megakaryocyte progenitors. In addition to DNA binding, distinct N-terminal regions, including residues in the first 84 amino acids, promote platelet release and restrict megakaryocyte growth. In contrast, abrogation of GATA1-FOG-1 interaction leads to loss of differentiation, but growth of blocked immature megakaryocytes is controlled. Thus, distinct GATA1 domains regulate terminal megakaryocyte gene expression leading to platelet release and restrain megakaryocyte growth, and these processes can be uncoupled.


Genomic profiles of Indonesian colorectal cancer patients.
Background: Colorectal cancer (CRC) is one of the most commonly diagnosed cancers worldwide and genetic mutation plays a vital role in CRC development. A previous study has suggested that genetic alterations among Indonesian patients with CRC might differ from those known in developed countries. This study aimed to describe the genomic profiles of Indonesian patients with CRC. Methods: A total of 13 patients were recruited for this study from May to July 2019. Tissue samples were collected, and genomic DNA was extracted from the samples. AmpliSeq for Illumina Cancer HotSpot Panel v2 Next-generation sequencing was used for DNA sequencing and a genome analysis toolkit was used for local realignment around the discovered variants. Results: A total of 45 genes comprising 391 single nucleotide variants (SNVs) with a depth >10 were observed. The genes with the most variants were STK11, SMAD4, EGFR, and ERBB4 and the genes with the most non-synonymous variants were SMAD4, TP53, FGFR3, CDKN2A, and STK11. Genes and SNVs in at least 90% of all samples consisted of 43 genes comprising 286 variants. Genes with the most non-synonymous SNVs were EGFR, SMO, FGFR3, TP53, STK11, CDKN2A. Genes related to the chromosomal instability pathway, such as TP53, SMAD4, KRAS, and APC, are also found in the analysis. Conclusions: Our findings showed that all patients with CRC in this study had genetic mutations in the chromosomal instability pathway. Analysis of genetic mutation of Indonesian patients with CRC might be crucial for advanced targeted therapy and for better clinical outcomes.


EPID dosimetry: effect of different layers of materials on absorbed dose response.
Commercial EPIDs are normally used in indirect detection mode (iEPID) where incident x-ray photons are converted to optical photons in a phosphor scintillator, which are then detected by a photodiode array. The EPIDs are constructed from a number of nonwater equivalent materials which affect the dose response of the detector. The so-called direct detection EPIDs (dEPIDs), operating without the phosphor layer, have been reported to display dose response close to in-water data. In this study, the effect that different layers of materials in the EPID have on the dose response was experimentally investigated and evaluated with respect to changes in field size response and beam profiles. An iEPID was disassembled and the different layers of materials were removed or replaced with other materials. Data were also obtained on and off the support arm and with a sheet of opaque paper blocking the optical photons from the gadolinium oxysulfide (Gd2S2O:Tb) phosphor layer. Field size response was measured for field sizes ranging from 2 x 2 to 25 x 25 cm2, and profiles for the 25 x 25 cm2 beams were extracted from the data. The iEPID configuration was found to be very sensitive to backscatter. The increases in output with solid water backscatter compared to the no backscatter case were 14.7% and 6.6% at the largest field size investigated for the 6 and 18 MV beams, respectively. The Gd2S2O:Tb phosphor layer had a large influence on field size response as well as beam profiles for 6 MV photons, while no major effects were observed for the 18 MV beam. For 18 MV large differences in dose response were found when the standard 1 mm Cu buildup was changed for dmax equivalent Cu or solid water buildup, indicating that head scatter largely influences dose response for this energy. When the optical photons originating in the Gd2S2O:Tb layer were blocked from reaching the photodiodes, both field size output data and beam profiles corresponded well with data obtained in the dEPID configuration as well as reference ion chamber data for both energies. As expected, changing the layers of material in the EPID had a dramatic effect on dose response, which was often quite complex. For 6 MV, the complex dose response is mainly caused by the optical photons from the Gd2S2O:Tb layer, while insufficient filtering of scattered radiation largely affects the dose response for the 18 MV beam. The iEPID was also found to be very sensitive to backscatter for both energies. Blocking the optical photons created in the Gd2S2O:Tb layer essentially changed the iEPID configuration into the dEPID configuration, thus demonstrating great potential for a system that can be optimized for both imaging and dosimetry.


Training Family Doctors and Primary Care Nurses in Evidence-based Prevention, Screening and Management of Cardiovascular Risks in Western Ukraine: A Longitudinal Study.
The Ukrainian primary healthcare programme of preventive and screening recommendations has not been evidence-based. The traditional system of continuous medical education in Ukraine places participants in the role of passive listeners. This study explored the effects of an interactive training course on evidence-based prevention and screening of cardiovascular risks, on changes in Ukrainian family doctors' (FDs) and primary care nurses' (PCNs) knowledge and readiness to change practice over time. Three hundred and seven FDs and PCNs participated in the study. Changes in participants' knowledge were assessed with 20 multiple choice questions, and their readiness to change practice with a five-item questionnaire. These were administered before, immediately after, three and twelve months after training. The mean pre-course knowledge score was 6.1 (SD 1.8) out of 20, increasing to 14.9 (SD 2.3) immediately afterwards (p<0.001). Three months later it was 10.2 (SD 3.2) and at one year it was 10.4 (SD 3.3), both of which were significantly higher than the pre-training level (p<0.005). The percentage of participants that were highly motivated to change their practice increased from 18.4% before the training to 62.3% immediately afterwards (p<0.001). Three months later, this fell to 40.4%. At 12 months it further reduced to 27.4%, but was still significantly higher than the baseline level (p<0.001). The interactive training was effective in increasing both participants' knowledge and their readiness to change their clinical practice. The impact of the training diminished over time, but was still evident a year later.


Bevacizumab for Patients with Recurrent Gliomas Presenting with a Gliomatosis Cerebri Growth Pattern.
Bevacizumab has been shown to improve progression-free survival and neurologic function, but failed to improve overall survival in newly diagnosed glioblastoma and at first recurrence. Nonetheless, bevacizumab is widely used in patients with recurrent glioma. However, its use in patients with gliomas showing a gliomatosis cerebri growth pattern is contentious. Due to the marked diffuse and infiltrative growth with less angiogenic tumor growth, it may appear questionable whether bevacizumab can have a therapeutic effect in those patients. However, the development of nodular, necrotic, and/or contrast-enhancing lesions in patients with a gliomatosis cerebri growth pattern is not uncommon and may indicate focal neo-angiogenesis. Therefore, control of growth of these lesions as well as control of edema and reduction of steroid use may be regarded as rationales for the use of bevacizumab in these patients. In this retrospective patient series, we report on 17 patients with primary brain tumors displaying a gliomatosis cerebri growth pattern (including seven glioblastomas, two anaplastic astrocytomas, one anaplastic oligodendroglioma, and seven diffuse astrocytomas). Patients have been treated with bevacizumab alone or in combination with lomustine or irinotecan. Seventeen matched patients treated with bevacizumab for gliomas with a classical growth pattern served as a control cohort. Response rate, progression-free survival, and overall survival were similar in both groups. Based on these results, anti-angiogenic therapy with bevacizumab should also be considered in patients suffering from gliomas with a mainly infiltrative phenotype.


Breastfeeding support - the importance of self-efficacy for low-income women.
Breastfeeding is a key determinant in promoting public health and reducing health inequality. Low-income women have a significantly lower level of breastfeeding. Midwives in the UK have been encouraged to implement the World Health Organization/United Nations Children's Fund's Ten Steps to Successful Breastfeeding, but to date, there has been no evaluation of the impact of the training initiative on the breastfeeding behaviours of low-income women. As part of a wider study, this qualitative component was designed to answer the question - what are the views and experiences of low-income women (defined by Jarman scores) in relation to their breastfeeding support received in the post-natal period? A sample of seven women was interviewed. The in-depth interviews were analysed using a qualitative, thematic approach based on the self-efficacy theory. The four themes that emerged from the data were the following: breastfeeding related to the woman's self-confidence, the social environment in which the woman lived, knowledge of breastfeeding and the influence of maternity services on breastfeeding outcomes. These themes were interpreted in relation to the self-efficacy theory. The findings suggest that the components that inform self-efficacy are consistent with the themes from the data, suggesting that midwives and other health professionals should take the psychosocial aspects of breastfeeding support into account. As this important feature of breastfeeding support is not explicitly part of the current Ten Steps to Successful Breastfeeding, we suggest that further research and debate could inform expansion of these minimum standards to include the psychosocial aspects.


Toxicities Caused by Head and Neck Cancer Treatments and Their Influence on the Development of Malnutrition: Review of the Literature.
Malnutrition poses a significant problem for oncology patients, resulting in fatalities within this population. Patients with head and neck cancer (HNC) are at high risk, with up to 90% developing malnutrition. Common treatments used for HNC can often lead to adverse side effects, including oral health conditions, gastrointestinal upsets, and several metabolic changes. Consequently, treatments can cause inadequate nutritional intake, resulting in a reduction in energy consumption, and alterations in energy utilization, contributing to the development of malnutrition. Furthermore, the presence of these treatment toxicities, and the related malnutrition can lead to reduced quality of life, weight loss, and psychological distress. There are interventions available (nutritional, medicinal, and physical therapies) that have demonstrated potential effectiveness in reducing the severity of symptomatic toxicities, reducing the risk of malnutrition, and improving survival outcomes of patients with HNC. Based on the findings of this review, there is an urgent need for the implementation or continuation of multi-disciplinary strategies, as well as updated and improved guidelines to assist in the prevention and treatment of malnutrition caused by treatment-related toxicities in patients with HNC.


Seven BMPs and all their receptors are simultaneously expressed in osteosarcoma cells.
Members of the bone morphogenetic protein (BMP) family and their receptors (BMPRs and activin receptors-ActRs) promote the development of bones with a fine regulation of their expression. Mutations in BMPs or BMPRs cause several diseases, as shown in knockout mice, such as skeletal defects, familial primary pulmonary hypertension and neoplasias. Osteosarcoma is the most frequent primary malignant tumor of bone. Due to their importance in bone development, BMPs, BMPRs and ActRs could also play a role in osteosarcoma growth and development. Previous data have shown that the overexpression of the BMPR-II was related to poor prognosis in malignant and metastatic bone tumors. We evaluate by reverse transcription-linked polymerase chain reaction analysis (RT-PCR) the expression pattern of BMPs, BMPRs and ActRs in five different human osteosarcoma cell lines (MG63, G292, HOS, SaOS and U2). Moreover, we performed the mutational screening of the complete BMPR-II mRNA by automated sequencing of the correspondent cDNA to evaluate the presence of point mutations in osteosarcoma cell lines. All the osteosarcoma cell lines studied simultaneously expressed the BMPs, BMPRs and ActRs investigated. No mutations were detected in the BMPR-II cDNA. Our results suggest the presence of a mechanism involving the simultaneous activation of the BMPs and their receptors in osteosarcoma cell lines.


Occupational and leisure time physical activity in contrasting relation to ambulatory blood pressure.
While moderate and vigorous leisure time physical activities are well documented to decrease the risk for cardiovascular disease, several studies have demonstrated an increased risk for cardiovascular disease in workers with high occupational activity. Research on the underlying causes to the contrasting effects of occupational and leisure time physical activity on cardiovascular health is lacking. The aim of this study was to examine the relation of objective and self-report measures of occupational and leisure time physical activity with 24-h ambulatory systolic blood pressure (BP). Results for self-reported physical activity are based on observations in 182 workers (60% male, mean age 51 years), while valid objective physical activity data were available in 151 participants. The usual level of physical activity was assessed by 5 items from the Job Content Questionnaire (high physical effort, lifting heavy loads, rapid physical activity, awkward body positions and awkward positions of head or arms at work) and one item asking about the general level of physical activity during non-working time. On a regular working day, participants wore an ambulatory BP monitor and an accelerometer physical activity monitor during 24 h. Associations were examined by means of Analysis of Covariance. Workers with an overall high level of self-reported occupational physical activity as well as those who reported to often lift heavy loads at work had a higher mean systolic BP at work, at home and during sleep. However, no associations were observed between objectively measured occupational physical activity and BP. In contrast, those with objectively measured high proportion of moderate and vigorous leisure time physical activity had a significantly lower mean systolic BP during daytime, while no differences were observed according to self-reported level of leisure time physical activity. These findings suggest that workers reporting static occupational physical activities, unlike general physically demanding tasks characterized by dynamic movements of large muscle groups, are related to a higher daily systolic BP, while high objective levels of moderate and vigorous leisure time physical activity are related to lower daytime systolic BP. Ambulatory systolic BP may be a physiological explanatory factor for the contrasting effects of occupational and leisure time physical activity.


The fate of siderophores: antagonistic environmental interactions in exudate-mediated micronutrient uptake.
Organisms acquire metals from the environment by releasing small molecules that solubilize and promote their specific uptake. The best known example of this nutrient uptake strategy is the exudation of siderophores, which are a structurally-diverse class of molecules that are traditionally viewed as being integral to iron uptake. Siderophores have been proposed to act through a variety of processes, but their effectiveness can be mitigated by a variety of chemical and physical processes of both biotic and abiotic origin. Processes that occur at the surface of minerals can degrade or sequester siderophores, preventing them from fulfilling their function of returning metals to the organism. In addition, biotic processes including enzymatic degradation of the siderophore and piracy of the metal or of the siderophore complex also disrupt iron uptake. Some organisms have adapted their nutrient acquisition strategies to address these potential pitfalls, producing multiple siderophores and other exudates that take advantage of varying kinetic and thermodynamic factors to allow the continued uptake of metals. A complete understanding of the factors that contribute to metal uptake in nature will require a concerted effort to study processes identified in laboratory systems in the context of more complicated environmental systems.


Two-dimensional TIRF-SIM-traction force microscopy (2D TIRF-SIM-TFM).
Quantifying small, rapidly evolving forces generated by cells is a major challenge for the understanding of biomechanics and mechanobiology in health and disease. Traction force microscopy remains one of the most broadly applied force probing technologies but typically restricts itself to slow events over seconds and micron-scale displacements. Here, we improve >2-fold spatially and >10-fold temporally the resolution of planar cellular force probing compared to its related conventional modalities by combining fast two-dimensional total internal reflection fluorescence super-resolution structured illumination microscopy and traction force microscopy. This live-cell 2D TIRF-SIM-TFM methodology offers a combination of spatio-temporal resolution enhancement relevant to forces on the nano- and sub-second scales, opening up new aspects of mechanobiology to analysis.


Synthesis, Morphological, and Electro-optical Characterizations of Metal/Semiconductor Nanowire Heterostructures.
In this letter, we demonstrate the formation of unique Ga/GaAs/Si nanowire heterostructures, which were successfully implemented in nanoscale light-emitting devices with visible room temperature electroluminescence. Based on our recent approach for the integration of InAs/Si heterostructures into Si nanowires by ion implantation and flash lamp annealing, we developed a routine that has proven to be suitable for the monolithic integration of GaAs nanocrystallite segments into the core of silicon nanowires. The formation of a Ga segment adjacent to longer GaAs nanocrystallites resulted in Schottky-diode-like I/V characteristics with distinct electroluminescence originating from the GaAs nanocrystallite for the nanowire device operated in the reverse breakdown regime. The observed electroluminescence was ascribed to radiative band-to-band recombinations resulting in distinct emission peaks and a low contribution due to intraband transition, which were also observed under forward bias. Simulations of the obtained nanowire heterostructure confirmed the proposed impact ionization process responsible for hot carrier luminescence. This approach may enable a new route for on-chip photonic devices used for light emission or detection purposes.


Safety and efficacy of combination therapy of interferon-α2 and ruxolitinib in polycythemia vera and myelofibrosis.
Interferon-α2 reduces elevated blood cell counts and splenomegaly in patients with myeloproliferative neoplasms (MPN) and may restore polyclonal hematopoiesis. Its use is limited by inflammation-mediated toxicity, leading to treatment discontinuation in 10-30% of patients. Ruxolitinib, a potent anti-inflammatory agent, has demonstrated benefit in myelofibrosis (MF) and polycythemia vera (PV) patients. Combination therapy (CT) with these two agents may be more efficacious than monotherapy with either, potentially improving tolerability of interferon-α2 as well. We report the preliminary results from a phase II study of CT with pegylated interferon-α2 and ruxolitinib in 50 MPN patients (PV, n = 32; low-/intermediate-1-risk MF, n = 18), the majority (n = 47) being resistant and/or intolerant to interferon-α2 monotherapy. Objectives included remission (2013 revised criteria encompassing histologic, hematologic, and clinical responses), complete hematologic response (CHR), molecular response, and toxicity. Follow-up was 12 months. Partial remission (PR) and sustained CHR were achieved in 9% and 44% of PV patients, respectively. In MF patients, complete or partial remission was achieved in 39%, and sustained CHR in 58%. The median JAK2V617F allele burden declined significantly in both groups. Hematologic toxicity was the most common adverse event and was managed by dose reduction. Thirty-seven serious adverse events were recorded in 23 patients; the discontinuation rate was 20%. We conclude that CT with interferon-α2 and ruxolitinib is efficacious in patients with low-/intermediate-1-risk MF and, to a lesser extent, in patients with PV. These preliminary results encourage phase III studies as well as a study with CT in newly diagnosed MPN patients.


Household food access and child malnutrition: results from the eight-country MAL-ED study.
Stunting results from decreased food intake, poor diet quality, and a high burden of early childhood infections, and contributes to significant morbidity and mortality worldwide. Although food insecurity is an important determinant of child nutrition, including stunting, development of universal measures has been challenging due to cumbersome nutritional questionnaires and concerns about lack of comparability across populations. We investigate the relationship between household food access, one component of food security, and indicators of nutritional status in early childhood across eight country sites. We administered a socioeconomic survey to 800 households in research sites in eight countries, including a recently validated nine-item food access insecurity questionnaire, and obtained anthropometric measurements from children aged 24 to 60 months. We used multivariable regression models to assess the relationship between household food access insecurity and anthropometry in children, and we assessed the invariance of that relationship across country sites. Average age of study children was 41 months. Mean food access insecurity score (range: 0-27) was 5.8, and varied from 2.4 in Nepal to 8.3 in Pakistan. Across sites, the prevalence of stunting (42%) was much higher than the prevalence of wasting (6%). In pooled regression analyses, a 10-point increase in food access insecurity score was associated with a 0.20 SD decrease in height-for-age Z score (95% CI 0.05 to 0.34 SD; p = 0.008). A likelihood ratio test for heterogeneity revealed that this relationship was consistent across countries (p = 0.17). Our study provides evidence of the validity of using a simple household food access insecurity score to investigate the etiology of childhood growth faltering across diverse geographic settings. Such a measure could be used to direct interventions by identifying children at risk of illness and death related to malnutrition.


Extracellular histones are major mediators of death in sepsis.
Hyperinflammatory responses can lead to a variety of diseases, including sepsis. We now report that extracellular histones released in response to inflammatory challenge contribute to endothelial dysfunction, organ failure and death during sepsis. They can be targeted pharmacologically by antibody to histone or by activated protein C (APC). Antibody to histone reduced the mortality of mice in lipopolysaccharide (LPS), tumor necrosis factor (TNF) or cecal ligation and puncture models of sepsis. Extracellular histones are cytotoxic toward endothelium in vitro and are lethal in mice. In vivo, histone administration resulted in neutrophil margination, vacuolated endothelium, intra-alveolar hemorrhage and macro- and microvascular thrombosis. We detected histone in the circulation of baboons challenged with Escherichia coli, and the increase in histone levels was accompanied by the onset of renal dysfunction. APC cleaves histones and reduces their cytotoxicity. Co-infusion of APC with E. coli in baboons or histones in mice prevented lethality. Blockade of protein C activation exacerbated sublethal LPS challenge into lethality, which was reversed by treatment with antibody to histone. We conclude that extracellular histones are potential molecular targets for therapeutics for sepsis and other inflammatory diseases.


Association between six-minute walk test parameters and the health-related quality of life in patients with pulmonary Mycobacterium avium complex disease.
Pulmonary Mycobacterium avium complex (pMAC) disease is a chronic, slowly progressive disease. The aim of the present study was to determine the association of six-minute walk test (6MWT) parameters with pulmonary function and the health-related quality of life (HRQL) in patients with pMAC disease. This cross-sectional study included adult patients with pMAC and was conducted at Keio University Hospital. We investigated the relationship of 6MWT parameters with clinical parameters, including pulmonary function, and HRQL, which was assessed using the 36-Item Short Form Health Survey (SF-36) and St. George's Respiratory Questionnaire (SGRQ). In total, 103 consecutive patients with pMAC participated in 6MWT (median age, 64 years; 80 women) and completed SF-36 and SGRQ. The six-minute walk distance (6MWD) showed significant negative and positive correlations with all SGRQ domain scores [ρ = (- 0.54)-(- 0.32)] and the physical component summary (PCS) score (ρ = 0.39) in SF-36, respectively; the opposite was observed for the final Borg scale (FBS) score (all SGRQ scores, ρ = 0.34-0.58; PCS score, ρ = - 0.50). The distance-saturation product showed significant negative and positive correlations with all SGRQ scores [ρ = (- 0.29)-(- 0.55)] and the PCS score (ρ = 0.40), respectively. Multivariate analysis revealed that 6MWD and the FBS score were significant predictors of HRQL. Our findings suggest that 6MWD and the FBS score are useful parameters for evaluating HRQL in patients with pMAC. Further studies should investigate the impact of 6WMT parameters on disease progression, treatment responses, and prognosis.


Surgery for pleural mesothelioma, when it is indicated and why: arguments against surgery for malignant pleural mesothelioma.
Extrapleural pneumonectomy (EPP) and pleurectomy decortication (PD) are radical operations for malignant pleural mesothelioma (MPM) that remain controversial among thoracic surgeons. There is a lack of randomized evidence to support a survival benefit when major surgical resection is included in multi-modality treatment regimens. Current data from retrospective single institution reviews and prospective trials such as the Surgery for Mesothelioma After Radiation Therapy (SMART) trial are limited by biased patient selection to include only the healthiest patients with most limited disease burden. This patient population predictably has relatively longer survival times than patients with inoperable advanced disease. The only randomized trial to date that has objectively evaluated the true benefit of surgical resection was the Mesothelioma and Radical Surgery (MARS) trial which actually showed shorter survival times among patients who underwent EPP compared with those treated medically. Critics of the MARS trial cite a high perioperative mortality rate for driving these results, however a similar trial has never been repeated to refute the MARS trial results. Finally, it is relevant to consider the high mortality and morbidity rates associated with major operations when recommending these interventions to MPM patients. There is a growing body of literature that identifies patients who clearly obtain no benefit from surgery including those with sarcomatoid or biphasic histology, nodal disease, elevated CRP, elevated platelets and advanced age. Surgery in MPM has risks and is of questionable benefit with outcomes data biased by patient selection of those who will have longer overall survival times regardless of treatment.


13C ENDOR Spectroscopy of Lipoxygenase-Substrate Complexes Reveals the Structural Basis for C-H Activation by Tunneling.
In enzymatic C-H activation by hydrogen tunneling, reduced barrier width is important for efficient hydrogen wave function overlap during catalysis. For native enzymes displaying nonadiabatic tunneling, the dominant reactive hydrogen donor-acceptor distance (DAD) is typically ca. 2.7 Å, considerably shorter than normal van der Waals distances. Without a ground state substrate-bound structure for the prototypical nonadiabatic tunneling system, soybean lipoxygenase (SLO), it has remained unclear whether the requisite close tunneling distance occurs through an unusual ground state active site arrangement or by thermally sampling conformational substates. Herein, we introduce Mn2+ as a spin-probe surrogate for the SLO Fe ion; X-ray diffraction shows Mn-SLO is structurally faithful to the native enzyme. 13C ENDOR then reveals the locations of 13C10 and reactive 13C11 of linoleic acid relative to the metal; 1H ENDOR and molecular dynamics simulations of the fully solvated SLO model using ENDOR-derived restraints give additional metrical information. The resulting three-dimensional representation of the SLO active site ground state contains a reactive (a) conformer with hydrogen DAD of ∼3.1 Å, approximately van der Waals contact, plus an inactive (b) conformer with even longer DAD, establishing that stochastic conformational sampling is required to achieve reactive tunneling geometries. Tunneling-impaired SLO variants show increased DADs and variations in substrate positioning and rigidity, confirming previous kinetic and theoretical predictions of such behavior. Overall, this investigation highlights the (i) predictive power of nonadiabatic quantum treatments of proton-coupled electron transfer in SLO and (ii) sensitivity of ENDOR probes to test, detect, and corroborate kinetically predicted trends in active site reactivity and to reveal unexpected features of active site architecture.


Knockdown of Nrf2 inhibits the angiogenesis of rat cardiac micro-vascular endothelial cells under hypoxic conditions.
Angiogenesis plays an important role in myocardial repair after myocardial infarction (MI). Cardiac micro-vascular endothelial cells (CMECs) are important participants in myocardial angiogenesis processes. Recent studies have revealed that Nuclear factor-erythroid 2-related factor 2 (Nrf2), a master transcription factor of endogenous anti-oxidative defense systems, exerts cardio-protection in the cardiovascular system. However, the role of Nrf2 in the process of myocardial angiogenesis and corresponding mechanisms are not fully understood. Thus, the present study investigated the role of Nrf2 in the angiogenesis of rat CMECs to hypoxia. Trans-well assay, three-dimensional Matrigel assay were used to determine cell migration and vascular tube formation. Real-time RT-PCR, ELISA and Western blot were measured mRNA and protein expression. Here, we report that the mRNA and protein expression of Nrf2 and heme oxygenase-1(HO-1) were temporarily upregulated under hypoxic condition. Furthermore, knock down of Nrf2 significantly suppressed the migration and vascular tube formation of rat CMECs to hypoxia, Nrf2 knockdown also significantly decreased HO-1 and vascular endothelial growth factor (VEGF) expression at 48 h after transfection under hypoxic condition. Finally, transfection of CMECs with the Nrf2 over-expressing lentiviral vector upregulated HO-1 expression with a concomitant increase in cell migration and vascular tube formation induced by hypoxia, and this effect was greatly attenuated in the presence of ZnPP (a HO-1 inhibitor). Taken together, these results suggest that Nrf2 may mediate the angiogenesis of CMECs under hypoxic condition, and HO-1 is involved in regulating the angiogenesis of CMECs through Nrf2. Therefore, Nrf2 is a potent regulator of hypoxia-condition mediated angiogenesis in CMECs, which may provide a therapeutic strategy for myocardial repair after MI.


Diode-laser-to-waveguide butt coupling.
A diode-laser-to-waveguide butt-coupling model is described. The model takes into consideration the Fabry-Perot reflection and transmission of the étalon formed by the laser front facet and the waveguide entrance facet. The model predicts coupling efficiency and the coupled-power fluctuations that occur versus the separation between the laser and the waveguide. Calculations performed for Fabry-Perot-laser-to-KTP waveguide coupling show that the transverse and the angular alignment tolerances of the waveguide can be increased when the waveguide entrance facet is antireflection coated. The longitudinal alignment tolerance of the waveguide can be increased by use of an index-matching gel between the laser and the waveguide.


Spatial distribution of seroprevalence for Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia canis, and Dirofilaria immitis in dogs in Washington, Oregon, and California.
In the US little spatially defined information regarding exposure to most vector-borne pathogens in dogs is available for the states of California (CA), Oregon (OR), and Washington (WA). The purpose of the present study was to evaluate the spatial distribution of seroprevalence for 4 vector-borne pathogens, Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia canis, and Dirofilaria immitis, across the 3 western coastal states of the contiguous United States that extend from the northern Mexican to the southern Canadian border. A convenience sample, targeting blood from 20 pet dogs per county across CA, OR, and WA, was evaluated using a canine point-of-care ELISA kit. Geographic coordinates of home zip code were displayed using a geographic information system. A total of 2431 dogs from CA, OR, and WA were tested. The overall seroprevalence was highest for A. phagocytophilum (2.4%), followed by B. burgdorferi (1.2%), and E. canis (0.7%). The prevalence of infection with D. immitis was 0.7%. At the individual dog level, there was a significant association between seropositivity to B. burgdorferi and A. phagocytophilum (odds ratio=18.7, 95% confidence interval=6.8-47.1). For most positive results, prevalence tended to decrease with increasing latitude; thus, the highest rates of seropositivity occurred in CA, followed by OR, and then WA; one exception was seropositivity for B. burgdorferi, which was higher in WA (0.38%) than in OR (0.15%), but considerably lower than in CA (2.00%). In WA, dogs that tested positive for A. phagocytophilum, E. canis, and B. burgdorferi were in the southern Puget Sound area. For D. immitis, none of the dogs in WA was positive. Seropositivity for vector-borne pathogens is broadly but patchily distributed in dogs in CA, OR, and WA.


Facilitative glucose transporter Glut1 is actively excluded from rod outer segments.
Photoreceptors are among the most metabolically active cells in the body, relying on both oxidative phosphorylation and glycolysis to satisfy their high energy needs. Local glycolysis is thought to be particularly crucial in supporting the function of the photoreceptor's light-sensitive outer segment compartment, which is devoid of mitochondria. Accordingly, it has been commonly accepted that the facilitative glucose transporter Glut1 responsible for glucose entry into photoreceptors is localized in part to the outer segment plasma membrane. However, we now demonstrate that Glut1 is entirely absent from the rod outer segment and is actively excluded from this compartment by targeting information present in its cytosolic C-terminal tail. Our data indicate that glucose metabolized in the outer segment must first enter through other parts of the photoreceptor cell. Consequently, the entire energy supply of the outer segment is dependent on diffusion of energy-rich substrates through the thin connecting cilium that links this compartment to the rest of the cell.


High-resolution structure of a Na+/H+ antiporter dimer obtained by pulsed electron paramagnetic resonance distance measurements.
Transient or partial formation of complexes between biomacromolecules is a general mechanism used to control cellular functions. Several of these complexes escape structure determination by crystallographic means. We developed a new approach for determining the structure of protein dimers in the native environment (e.g., in the membrane) with high resolution in cases where the structure of the two monomers is known. The approach is based on measurements of distance distributions between spin labels in the range between 2 and 6 nanometers by a pulsed electron paramagnetic resonance technique and explicit modeling of spin label conformations. By applying this method to the membrane protein homodimer of the Na(+)/H(+) antiporter NhaA of Escherichia coli, the structure of the presumably physiological dimer was determined. It reveals two points of contact between the two monomers, with one of them confirming results of earlier cross-linking experiments.


Calcium-based nanoparticles accelerate skin wound healing.
Nanoparticles (NPs) are small entities that consist of a hydroxyapatite core, which can bind ions, proteins, and other organic molecules from the surrounding environment. These small conglomerations can influence environmental calcium levels and have the potential to modulate calcium homeostasis in vivo. Nanoparticles have been associated with various calcium-mediated disease processes, such as atherosclerosis and kidney stone formation. We hypothesized that nanoparticles could have an effect on other calcium-regulated processes, such as wound healing. In the present study, we synthesized pH-sensitive calcium-based nanoparticles and investigated their ability to enhance cutaneous wound repair. Different populations of nanoparticles were synthesized on collagen-coated plates under various growth conditions. Bilateral dorsal cutaneous wounds were made on 8-week-old female Balb/c mice. Nanoparticles were then either administered intravenously or applied topically to the wound bed. The rate of wound closure was quantified. Intravenously injected nanoparticles were tracked using a FLAG detection system. The effect of nanoparticles on fibroblast contraction and proliferation was assessed. A population of pH-sensitive calcium-based nanoparticles was identified. When intravenously administered, these nanoparticles acutely increased the rate of wound healing. Intravenously administered nanoparticles were localized to the wound site, as evidenced by FLAG staining. Nanoparticles increased fibroblast calcium uptake in vitro and caused contracture of a fibroblast populated collagen lattice in a dose-dependent manner. Nanoparticles also increased the rate of fibroblast proliferation. Intravenously administered, calcium-based nanoparticles can acutely decrease open wound size via contracture. We hypothesize that their contraction effect is mediated by the release of ionized calcium into the wound bed, which occurs when the pH-sensitive nanoparticles disintegrate in the acidic wound microenvironment. This is the first study to demonstrate that calcium-based nanoparticles can have a therapeutic benefit, which has important implications for the treatment of wounds.


Mortality in HIV-1-seropositive women, their spouses and their newly born children during 36 months of follow-up in Kinshasa, Zaïre.
To calculate 3-year mortality rates in HIV-1-seropositive and HIV-1-seronegative mothers, their newborn children and the fathers of these children. Longitudinal cohort study of HIV-1-seropositive, age and parity-matched HIV-1-seronegative pregnant women, their newborn babies and the fathers of these children. Obstetric ward and follow-up clinic at a large municipal hospital in Kinshasa, Zaïre. A total of 335 newborn children and their 327 HIV-1-seropositive mothers and 341 newborn children and their 337 HIV-1-seronegative mothers and the fathers of these children. Rates of vertical HIV-1 transmission and maternal, paternal and early childhood mortality. The lower and upper bounds of vertical transmission were 27 and 50%, respectively. The 3-year mortality rate was 44% in children with vertically acquired HIV-1 infection, 25% in children with HIV-1-seropositive mothers and indeterminant HIV-1 infection status, and 6% in uninfected children with HIV-1-seronegative mothers. HIV-1-seropositive women who transmitted HIV-1 infection to their most recently born child had lost a greater number of previously born children (mean, 1.5 versus 0.5; P < 0.05), were more likely to have had AIDS at delivery (25 versus 12%; P < 0.01) and were more likely to die during follow-up (22 versus 9%; P < 0.01) than HIV-1-seropositive women who did not transmit HIV-1 infection to their newborn child. Twenty-five out of 239 (10.4%) fathers of children with HIV-1-seropositive mothers, not lost to follow-up, died compared with three out of 310 (1%) fathers of children with HIV-1-seronegative mothers (P < 0.01). Families in Kinshasa, Zaïre, in which the mother was HIV-1-seropositive experienced a five to 10-fold higher maternal, paternal and early childhood mortality rate than families in which the mother was HIV-1-seronegative.


The impact of global health initiatives on trust in health care provision under extreme resource scarcity: presenting an agenda for debate from a case study of emergency obstetric care in Northern Tanzania.
Through the nearly three decades that have passed since the Alma Ata conference on Primary Health Care, a wide range of global health initiatives and ideas have been advocated to improve the health of people living in developing countries. The issues raised in the Primary Health Care concept, the Structural Adjustment Programmes and the Health Sector Reforms have all influenced health service delivery. Increasingly however, health systems in developing countries are being described as having collapsed Do the advocated frameworks contribute to this collapse through not adequately including population trust as a determinant of the revival of health services, or are they primarily designed to satisfy the values of other actors within the health care system? This article argues there is an urgent need to challenge common thinking on health care provision under extreme resource scarcity. This article sets out to discuss and analyze the described collapse of health services through a brief case study on provision of Emergency Obstetric Care in Northern Tanzania. The article argues that post the Alma Ata conference on Primary Health Care developments in global health initiatives have not been successful in incorporating population trust into the frameworks, instead focusing narrowly on expert-driven solutions through concepts such as prevention and interventions. The need for quantifiable results has pushed international policy makers and donors towards vertical programmes, intervention approaches, preventive services and quantity as the coverage parameter. Health systems have consequently been pushed away from generalized horizontal care, curative services and quality assurance, all important determinants of trust. Trust can be restored, and to further this objective a new framework is proposed placing generalized services and individual curative care in the centre of the health sector policy domain. Preventive services are important, but should increasingly be handled by other sectors in a service focused health care system. To facilitate such a shift in focus we should acknowledge that limited resources are available and accept the conflict between population demand and expert opinion, with the aim of providing legitimate, accountable and trustworthy services through fair, deliberative, dynamic and incremental processes. A discussion of the acceptable level of quality, given the available resources, can then be conducted. The article presents for debate that an increased focus on quality and accountability to secure trust is an important precondition for enabling the political commitment to mobilize necessary resources to the health sector.


Directed termination PCR: a one-step approach to mutation detection.
We describe a novel PCR-based method that allows the generation of nested termination fragments by integrating both selective DNA amplification and directed chain termination into a single PCR reaction. These termination fragments can be examined for sequence variation in either denaturing or non-denaturing polyacrylamide gels. This method provides a one-step and highly effective approach for the detection of both insertions/deletions and single base pair substitutions in sequences up to 1 kb in length.


Gold-clay nanocomposite colloids with liquid-crystalline and plasmonic properties.
Imparting liquid-crystal (LC) materials with the plasmonic properties of metal nanoparticles is actively pursued for applications. We achieved this goal by synthetizing gold nanoparticles onto clay nanosheets, leading to nematic nanocomposite suspensions. Optical observations and structural analysis show the growth of the gold nanoparticles without altering the LC properties of the nanosheets. These colloids display plasmonic structural colours and they can be aligned by an electric field, which is relevant for fundamental and materials chemistry of colloidal LC.


Col4a1 mutation in mice causes defects in vascular function and low blood pressure associated with reduced red blood cell volume.
Collagen type IV is the major structural component of the basement membrane and COL4A1 mutations cause adult small vessel disease, familial porencephaly and hereditary angiopathy with nephropathy aneurysm and cramps (HANAC) syndrome. Here, we show that animals with a Col4a1 missense mutation (Col4a1(+/Raw)) display focal detachment of the endothelium from the media and age-dependent defects in vascular function including a reduced response to nor-epinephrine. Age-dependent hypersensitivity to acetylcholine is abolished by inhibition of nitric oxide synthase (NOS) activity, indicating that Col4a1 mutations affect vasorelaxation mediated by endothelium-derived nitric oxide (NO). These defects are associated with a reduction in basal NOS activity and the development of heightened NO sensitivity of the smooth muscle. The vascular function defects are physiologically relevant as they maintain in part the hypotension in mutant animals, which is primarily associated with a reduced red blood cell volume due to a reduction in red blood cell number, rather than defects in kidney function. To understand the molecular mechanism underlying these vascular defects, we examined the deposition of collagen type IV in the basement membrane, and found it to be defective. Interestingly, this mutation also leads to activation of the unfolded protein response. In summary, our results indicate that mutations in COL4A1 result in a complex vascular phenotype encompassing defects in maintenance of vascular tone, endothelial cell function and blood pressure regulation.


Antimutator alleles of yeast DNA polymerase gamma modulate the balance between DNA synthesis and excision.
Mutations in mitochondrial DNA (mtDNA) are an important cause of disease and perhaps aging in human. DNA polymerase gamma (pol γ), the unique replicase inside mitochondria, plays a key role in the fidelity of mtDNA replication through selection of the correct nucleotide and 3'-5' exonuclease proofreading. For the first time, we have isolated and characterized antimutator alleles in the yeast pol γ (Mip1). These mip1 mutations, localised in the 3'-5' exonuclease and polymerase domains, elicit a 2-15 fold decrease in the frequency of mtDNA point mutations in an msh1-1 strain which is partially deficient in mtDNA mismatch-repair. In vitro experiments show that in all mutants the balance between DNA synthesis and exonucleolysis is shifted towards excision when compared to wild-type, suggesting that in vivo more opportunity is given to the editing function for removing the replicative errors. This results in partial compensation for the mismatch-repair defects and a decrease in mtDNA point mutation rate. However, in all mutants but one the antimutator trait is lost in the wild-type MSH1 background. Accordingly, the polymerases of selected mutants show reduced oligonucleotide primed M13 ssDNA synthesis and to a lesser extent DNA binding affinity, suggesting that in mismatch-repair proficient cells efficient DNA synthesis is required to reach optimal accuracy. In contrast, the Mip1-A256T polymerase, which displays wild-type like DNA synthesis activity, increases mtDNA replication fidelity in both MSH1 and msh1-1 backgrounds. Altogether, our data show that accuracy of wild-type Mip1 is probably not optimal and can be improved by specific (often conservative) amino acid substitutions that define a pol γ area including a loop of the palm subdomain, two residues near the ExoII motif and an exonuclease helix-coil-helix module in close vicinity to the polymerase domain. These elements modulate in a subtle manner the balance between DNA polymerization and excision.


Two routes to face perception: evidence from psychophysics and computational modeling.
The aim of this study was to separately analyze the role of featural and configural face representations. Stimuli containing only featural information were created by cutting the faces into their parts and scrambling them. Stimuli only containing configural information were created by blurring the faces. Employing an old-new recognition task, the aim of Experiments 1 and 2 was to investigate whether unfamiliar faces (Exp. 1) or familiar faces (Exp. 2) can be recognized if only featural or configural information is provided. Both scrambled and blurred faces could be recognized above chance level. A further aim of Experiments 1 and 2 was to investigate whether our method of creating configural and featural stimuli is valid. Pre-activation of one form of representation did not facilitate recognition of the other, neither for unfamiliar faces (Exp. 1) nor for familiar faces (Exp. 2). This indicates a high internal validity of our method for creating configural and featural face stimuli. Experiment 3 examined whether features placed in their correct categorical relational position but with distorted metrical distances facilitated recognition of unfamiliar faces. These faces were recognized no better than the scrambled faces in Experiment 1, providing further evidence that facial features are stored independently of configural information. From these results we conclude that both featural and configural information are important to recognize a face and argue for a dual-mode hypothesis of face processing. Using the psychophysical results as motivation, we propose a computational framework that implements featural and configural processing routes using an appearance-based representation based on local features and their spatial relations. In three computational experiments (Experiments 4-6) using the same sets of stimuli, we show how this framework is able to model the psychophysical data.


Incidence of thyroid cancer in England by ethnic group, 2001-2007.
Thyroid cancer incidence is increasing worldwide, but with large variations in incidence that may reflect either diagnostic bias or true ethnic differences. We sought to determine the effect of ethnicity on the incidence of thyroid cancer in England, a multiethnic population with a single health-care system. We analysed 11,263 thyroid cancer registrations with ethnicity obtained by linkage to the Hospital Episodes Statistics database. Incidence rate ratios (RRs) adjusted for age, sex and income were calculated for the six main non-White ethnic groups in England compared with Whites and to each other. Thyroid cancer incidence was higher in all ethnic groups, except Indians, compared with Whites: in Pakistanis (RR 1.79, 99% floating confidence interval (FCI) 1.47-2.19); Bangladeshis (RR 1.99, 99% FCI 1.46-2.71); Black Africans (RR 1.69, 99% FCI 1.34-2.13); Black Caribbeans (RR 1.56, 99% FCI 1.25-1.93); and Chinese (RR 2.14, 99% FCI 1.63-2.80). The risk of thyroid cancer in England varies significantly by ethnicity. The elevated incidence in most ethnic minorities is unlikely to be due to diagnostic bias and warrants further investigation.


Bromelain modulates T cell and B cell immune responses in vitro and in vivo.
The ability to modulate immune responses is a major aim of many vaccine and immunotherapeutic development programs. Bromelain, a mixture of cysteine proteases, modulates immunological responses and has been proposed to be of clinical use. However, the identity of the immune cells affected by bromelain and the specific cellular functions that are altered remain poorly understood. To address these shortcomings in our knowledge, we have used both in vitro and in vivo immunological assays to study the effects of bromelain. We found that bromelain enhanced T cell receptor (TCR) and anti-CD28-mediated T cell proliferation in splenocyte cultures by increasing the costimulatory activity of accessory cell populations. However, despite increased T cell proliferation, bromelain concomitantly decreased IL-2 production in splenocyte cultures. Additionally, bromelain did not affect TCR and CD28-induced proliferation of highly purified CD4+ T cells, but did inhibit IL-2 production by these cells. In vivo, bromelain enhanced T-cell-dependent, Ag-specific, B cell antibody responses. Again, bromelain induced a concomitant decrease in splenic IL-2 mRNA accumulation in immunized mice. Together, these data show that bromelain can simultaneously enhance and inhibit T cell responses in vitro and in vivo via a stimulatory action on accessory cells and a direct inhibitory action on T cells. This work provides important insights into the immunomodulatory activity of bromelain and has important implications for the use of exogenous cysteine proteases as vaccine adjuvants or immunomodulatory agents.