Transplanted bone marrow generates new neurons in human brains.
Adult bone marrow stem cells seem to differentiate into muscle, skin, liver, lung, and neuronal cells in rodents and have been shown to regenerate myocardium, hepatocytes, and skin and gastrointestinal epithelium in humans. Because we have demonstrated previously that transplanted bone marrow cells can enter the brain of mice and differentiate into neurons there, we decided to examine postmortem brain samples from females who had received bone marrow transplants from male donors. The underlying diseases of the patients were lymphocytic leukemia and genetic deficiency of the immune system, and they survived between 1 and 9 months after transplant. We used a combination of immunocytochemistry (utilizing neuron-specific antibodies) and fluorescent in situ hybridization histochemistry to search for Y chromosome-positive cells. In all four patients studied we found cells containing Y chromosomes in several brain regions. Most of them were nonneuronal (endothelial cells and cells in the white matter), but neurons were certainly labeled, especially in the hippocampus and cerebral cortex. The youngest patient (2 years old), who also lived the longest time after transplantation, had the greatest number of donor-derived neurons (7 in 10,000). The distribution of the labeled cells was not homogeneous. There were clusters of Y-positive cells, suggesting that single progenitor cells underwent clonal expansion and differentiation. We conclude that adult human bone marrow cells can enter the brain and generate neurons just as rodent cells do. Perhaps this phenomenon could be exploited to prevent the development or progression of neurodegenerative diseases or to repair tissue damaged by infarction or trauma.


Down-regulation of the CSLF6 gene results in decreased (1,3;1,4)-beta-D-glucan in endosperm of wheat.
(1,3;1,4)-beta-d-Glucan (beta-glucan) accounts for 20% of the total cell walls in the starchy endosperm of wheat (Triticum aestivum) and is an important source of dietary fiber for human nutrition with potential health benefits. Bioinformatic and array analyses of gene expression profiles in developing caryopses identified the CELLULOSE SYNTHASE-LIKE F6 (CSLF6) gene as encoding a putative beta-glucan synthase. RNA interference constructs were therefore designed to down-regulate CSLF6 gene expression and expressed in transgenic wheat under the control of a starchy endosperm-specific HMW subunit gene promoter. Analysis of wholemeal flours using an enzyme-based kit and by high-performance anion-exchange chromatography after digestion with lichenase showed decreases in total beta-glucan of between 30% and 52% and between 36% and 53%, respectively, in five transgenic lines compared to three control lines. The content of water-extractable beta-glucan was also reduced by about 50% in the transgenic lines, and the M(r) distribution of the fraction was decreased from an average of 79 to 85 x 10(4) g/mol in the controls and 36 to 57 x 10(4) g/mol in the transgenics. Immunolocalization of beta-glucan in semithin sections of mature and developing grains confirmed that the impact of the transgene was confined to the starchy endosperm with little or no effect on the aleurone or outer layers of the grain. The results confirm that the CSLF6 gene of wheat encodes a beta-glucan synthase and indicate that transgenic manipulation can be used to enhance the health benefits of wheat products.


VCP/p97 is essential for maturation of ubiquitin-containing autophagosomes and this function is impaired by mutations that cause IBMPFD.
VCP (VCP/p97) is a ubiquitously expressed member of the AAA(+)-ATPase family of chaperone-like proteins that regulates numerous cellular processes including chromatin decondensation, homotypic membrane fusion and ubiquitin-dependent protein degradation by the proteasome. Mutations in VCP cause a multisystem degenerative disease consisting of inclusion body myopathy, Paget disease of bone, and frontotemporal dementia (IBMPFD). Here we show that VCP is essential for autophagosome maturation. We generated cells stably expressing dual-tagged LC3 (mCherry-EGFP-LC3) which permit monitoring of autophagosome maturation. We determined that VCP deficiency by RNAi-mediated knockdown or overexpression of dominant-negative VCP results in significant accumulation of immature autophagic vesicles, some of which are abnormally large, acidified and exhibit cathepsin B activity. Furthermore, expression of disease-associated VCP mutants (R155H and A232E) also causes this autophagy defect. VCP was found to be essential to autophagosome maturation under basal conditions and in cells challenged by proteasome inhibition, but not in cells challenged by starvation, suggesting that VCP might be selectively required for autophagic degradation of ubiquitinated substrates. Indeed, a high percentage of the accumulated autophagic vesicles contain ubiquitin-positive contents, a feature that is not observed in autophagic vesicles that accumulate following starvation or treatment with Bafilomycin A. Finally, we show accumulation of numerous, large LAMP-1 and LAMP-2-positive vacuoles and accumulation of LC3-II in myoblasts derived from patients with IBMPFD. We conclude that VCP is essential for maturation of ubiquitin-containing autophagosomes and that defect in this function may contribute to IBMPFD pathogenesis.


Effects of parental smoking on exercise systolic blood pressure in adolescents.
In adults, exercise blood pressure seems to be more closely related to cardiovascular risk than resting blood pressure; however, few data are available on the effects of familial risk factors, including smoking habits, on exercise blood pressure in adolescents. Blood pressure at rest and during exercise, parental smoking, and other familial risk factors were investigated in 532 adolescents aged 12 to 17 years (14.6±1.5 years) in the Kiel EX.PRESS. (EXercise PRESSure) Study. Exercise blood pressure was determined at 1.5 W/kg body weight using a standardized submaximal cycle ergometer test. Mean resting blood pressure was 113.1±12.8/57.2±7.1 mm Hg, and exercise blood pressure was 149.9±19.8/54.2±8.6 mm Hg. Parental smoking increased exercise systolic blood pressure (+4.0 mm Hg, 3.1 to 4.9; P=0.03) but not resting blood pressure of the subjects (adjusted for age, sex, height, body mass index percentile, fitness). Parental overweight and familial hypertension were related to both higher resting and exercise systolic blood pressure values, whereas associations with an inactive lifestyle and a low educational level of the parents were found only with adolescents' blood pressure during exercise. The cumulative effect of familial risk factors on exercise systolic blood pressure was more pronounced than on blood pressure at rest. Parental smoking might be a novel risk factor for higher blood pressure, especially during exercise. In addition, systolic blood pressure during a submaximal exercise test was more closely associated with familial risk factors than was resting blood pressure, even in adolescents.


Integration of architectural and cytologic driven image algorithms for prostate adenocarcinoma identification.
The advent of digital slides offers new opportunities within the practice of pathology such as the use of image analysis techniques to facilitate computer aided diagnosis (CAD) solutions. Use of CAD holds promise to enable new levels of decision support and allow for additional layers of quality assurance and consistency in rendered diagnoses. However, the development and testing of prostate cancer CAD solutions requires a ground truth map of the cancer to enable the generation of receiver operator characteristic (ROC) curves. This requires a pathologist to annotate, or paint, each of the malignant glands in prostate cancer with an image editor software - a time consuming and exhaustive process. Recently, two CAD algorithms have been described: probabilistic pairwise Markov models (PPMM) and spatially-invariant vector quantization (SIVQ). Briefly, SIVQ operates as a highly sensitive and specific pattern matching algorithm, making it optimal for the identification of any epithelial morphology, whereas PPMM operates as a highly sensitive detector of malignant perturbations in glandular lumenal architecture. By recapitulating algorithmically how a pathologist reviews prostate tissue sections, we created an algorithmic cascade of PPMM and SIVQ algorithms as previously described by Doyle el al. [1] where PPMM identifies the glands with abnormal lumenal architecture, and this area is then screened by SIVQ to identify the epithelium. The performance of this algorithm cascade was assessed qualitatively (with the use of heatmaps) and quantitatively (with the use of ROC curves) and demonstrates greater performance in the identification of malignant prostatic epithelium. This ability to semi-autonomously paint nearly all the malignant epithelium of prostate cancer has immediate applications to future prostate cancer CAD development as a validated ground truth generator. In addition, such an approach has potential applications as a pre-screening/quality assurance tool.


Treatment of nonseptic olecranon bursitis. A controlled, blinded prospective trial.
We enrolled 42 patients with nonseptic olecranon bursitis in a double-blind prospective treatment trial to compare the efficacy of an intrabursal steroid preparation with that of an oral anti-inflammatory agent. Patients were randomized into one of four treatment regimens: (1) methylprednisolone acetate (20 mg) intrabursal injection and oral naproxen (1 g/d for 10 days), (2) methylprednisolone acetate (20 mg) intrabursal injection and oral placebo for 10 days, (3) oral naproxen (1 g/d for 10 days), and (4) oral placebo for 10 days. The degree of swelling in millimeters was assessed at study introduction and at 1, 3, and 6 weeks. At 6 months, the number of patients requiring reaspiration for bursitis recurrence was tabulated. Data at 1 week indicated that patients treated with an intrabursal methylprednisolone acetate injection (20 mg) demonstrated the most rapid decrease in swelling. At 6 weeks, the methylprednisolone-treated groups demonstrated sustained improvement. At 6 months, the mean number of reaspirations per patient for reaccumulation of bursal fluid was higher in groups 3 (1.0 +/- 1.2) and 4 (0.4 +/- 0.7). An intrabursal methylprednisolone acetate 20-mg injection seems to be the most effective treatment regimen for nonseptic olecranon bursitis.


Criteria for age estimation in living individuals.
This paper presents updated recommendations of the Study Group on Forensic Age Diagnostics for age estimations in living individuals in criminal proceedings. In order to increase the diagnostic accuracy and to improve the identification of age-relevant developmental disorders, a physical examination, an X-ray examination of the left hand, as well as a dental examination including the determination of the dental status and an X-ray of the dentition should be performed in each case. If the skeletal development of the hand is completed, an additional radiological examination of the clavicles should be carried out. Minimum requirements for reference studies are defined and recommendable studies are listed. Instructions for the examination and the preparation of expert reports are presented. The committee of the study group organizes annual proficiency tests for quality assurance.


Promoter targeting of chromatin-modifying complexes.
The action of multi-subunit complexes that are able to overcome the repressive effects of chromatin is an important step in the regulation of eukaryotic gene expression. Identification of complexes that modify the structure of chromatin to help factors access the underlying DNA has enhanced our understanding of how some genes are controlled. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) represent one group of complexes that regulate the level of acetylation on the N-terminal tails of core histone proteins. The SWI/SNF complex is the prototype of a second group of complexes, which use the energy of ATP-hydrolysis to alter histone-DNA contacts, leading to changes in chromatin conformation. Genetic studies in yeast have revealed that some of these multi-subunit complexes interact in vivo to control transcription of a subset of genes. It has become apparent that some gene promoters require modifications by both types of complexes. An important question regarding these two types of complexes is how they are recruited to the promoters of genes that are dependent on their activity for their expression. This review will tie together many studies on promoter recruitment of both HATs and SWI/SNF. Emphasis will be placed on recent data that demonstrates functional interplay between these two types of chromatin-remodeling activities. In addition, this review summarizes recent data demonstrating the ability of repressors and corepressors to recruit histone deacetylase complexes. Interestingly, many subunits of chromatin-modifying complexes in humans have been implicated in the development of cancer. Thus, studying how these complexes work can help us better understand human diseases.


BP180 (type XVII collagen) and its role in cutaneous biology and disease.
BP180 is a key component of the epidermal anchoring complex and functions to maintain adherence of the epidermis to the basement membrane. Structural studies have revealed that BP180 is a type II transmembrane protein with a long carboxy-terminal collagenous domain that projects into the extracellular region beneath the epidermal hemidesmosome. The collagenous domains have the characteristic tripeptide repeat, Gly-X-Y. A normal proteolytic processing event results in the shedding of the BP180 extracellular domain (LAD1) from the keratinocyte cell surface. The biologic relevance of this process is not yet known. The interactions of BP180 with other constituents of the anchoring complex have been extensively studied and underscore the importance of BP180 in the assembly and functioning of this cell-matrix adhesion structure. In addition to its role in maintaining the integrity of the dermal-epidermal junction, there is evidence that BP180 is involved in transmembrane signal transduction and in the regulation of keratinocyte differentiation. BP180 mutations are responsible for certain forms of JEB and a rare subform of epidermolysis bullosa simplex. In addition, 5 acquired blistering disorders (i.e. BP, HG, CP, LAD and LPP) are associated with an autoimmune response to BP180. In vivo and in vitro disease model systems have clearly established the pathogenic relevance of autoantibodies directed against specific sites on the BP180 extracellular domain. Molecular and cellular analyses of the autoimmune response in BP and HG have been unable to distinguish these 2 diseases, supporting the notion that HG can be considered a pregnancy-associated form of BP. In contrast, the anti-BP180 immune response of the other 3 disease--CP, LAD, and LPP--can be immunologically distinguished from BP and HG. The distinctions lie within the isotype and subclass of the autoantibodies, as well as in differences in their fine specificities or complement-fixing properties, or both. These differences are thought to account for the heterogeneous phenotypes observed in this family of autoimmune diseases.


NAD+ metabolism in health and disease.
Nicotinamide adenine dinucleotide (NAD(+)) is both a coenzyme for hydride-transfer enzymes and a substrate for NAD(+)-consuming enzymes, which include ADP-ribose transferases, poly(ADP-ribose) polymerases, cADP-ribose synthases and sirtuins. Recent results establish protective roles for NAD(+) that might be applicable therapeutically to prevent neurodegenerative conditions and to fight Candida glabrata infection. In addition, the contribution that NAD(+) metabolism makes to lifespan extension in model systems indicates that therapies to boost NAD(+) might promote some of the beneficial effects of calorie restriction. Nicotinamide riboside, the recently discovered nucleoside precursor of NAD(+) in eukaryotic systems, might have advantages as a therapy to elevate NAD(+) without inhibiting sirtuins, which is associated with high-dose nicotinamide, or incurring the unpleasant side-effects of high-dose nicotinic acid.


Human Depth Sensitivity Is Affected by Object Plausibility.
Using behavioral and fMRI paradigms, we asked how the physical plausibility of complex 3-D objects, as defined by the object's congruence with 3-D Euclidean geometry, affects behavioral thresholds and neural responses to depth information. Stimuli were disparity-defined geometric objects rendered as random dot stereograms, presented in plausible and implausible variations. In the behavior experiment, observers were asked to complete (1) a noise-based depth task that involved judging the depth position of a target embedded in noise and (2) a fine depth judgment task that involved discriminating the nearer of two consecutively presented targets. Interestingly, results indicated greater behavioral sensitivities of depth judgments for implausible versus plausible objects across both tasks. In the fMRI experiment, we measured fMRI responses concurrently with behavioral depth responses. Although univariate responses for depth judgments were largely similar across cortex regardless of object plausibility, multivariate representations for plausible and implausible objects were notably distinguishable along depth-relevant intermediate regions V3 and V3A, in addition to object-relevant LOC. Our data indicate significant modulations of both behavioral judgments of and neural responses to depth by object context. We conjecture that disparity mechanisms interact dynamically with the object recognition problem in the visual system such that disparity computations are adjusted based on object familiarity.


Sensing with magnetic dipolar resonances in semiconductor nanospheres.
In this work we propose two novel sensing principles of detection that exploit the magnetic dipolar Mie resonance in high-refractive-index dielectric nanospheres. In particular, we theoretically investigate the spectral evolution of the extinction and scattering cross sections of these nanospheres as a function of the refractive index of the external medium (next). Unlike resonances in plasmonic nanospheres, the spectral position of magnetic resonances in high-refractive-index nanospheres barely shifts as next changes. Nevertheless, there is a drastic reduction in the extinction cross section of the nanospheres when next increases, especially in the magnetic dipolar spectral region, which is accompanied with remarkable variations in the radiation patterns. Thanks to these changes, we propose two new sensing parameters, which are based on the detection of: i) the intensity variations in the transmitted or backscattered radiation by the dielectric nanospheres at the magnetic dipole resonant frequency, and ii) the changes in the radiation pattern at the frequency that satisfies Kerker's condition of near-zero forward radiation. To optimize the sensitivity, we consider several semiconductor materials and particles sizes.


Immune responses in healthy and allergic individuals are characterized by a fine balance between allergen-specific T regulatory 1 and T helper 2 cells.
The mechanisms by which immune responses to nonpathogenic environmental antigens lead to either allergy or nonharmful immunity are unknown. Single allergen-specific T cells constitute a very small fraction of the whole CD4+ T cell repertoire and can be isolated from the peripheral blood of humans according to their cytokine profile. Freshly purified interferon-gamma-, interleukin (IL)-4-, and IL-10-producing allergen-specific CD4+ T cells display characteristics of T helper cell (Th)1-, Th2-, and T regulatory (Tr)1-like cells, respectively. Tr1 cells consistently represent the dominant subset specific for common environmental allergens in healthy individuals; in contrast, there is a high frequency of allergen-specific IL-4-secreting T cells in allergic individuals. Tr1 cells use multiple suppressive mechanisms, IL-10 and TGF-beta as secreted cytokines, and cytotoxic T lymphocyte antigen 4 and programmed death 1 as surface molecules. Healthy and allergic individuals exhibit all three allergen-specific subsets in different proportions, indicating that a change in the dominant subset may lead to allergy development or recovery. Accordingly, blocking the suppressor activity of Tr1 cells or increasing Th2 cell frequency enhances allergen-specific Th2 cell activation ex vivo. These results indicate that the balance between allergen-specific Tr1 cells and Th2 cells may be decisive in the development of allergy.


Quantitative diagnosis of malignant pleural effusions by single-cell mechanophenotyping.
Biophysical characteristics of cells are attractive as potential diagnostic markers for cancer. Transformation of cell state or phenotype and the accompanying epigenetic, nuclear, and cytoplasmic modifications lead to measureable changes in cellular architecture. We recently introduced a technique called deformability cytometry (DC) that enables rapid mechanophenotyping of single cells in suspension at rates of 1000 cells/s-a throughput that is comparable to traditional flow cytometry. We applied this technique to diagnose malignant pleural effusions, in which disseminated tumor cells can be difficult to accurately identify by traditional cytology. An algorithmic diagnostic scoring system was developed on the basis of quantitative features of two-dimensional distributions of single-cell mechanophenotypes from 119 samples. The DC scoring system classified 63% of the samples into two high-confidence regimes with 100% positive predictive value or 100% negative predictive value, and achieved an area under the curve of 0.86. This performance is suitable for a prescreening role to focus cytopathologist analysis time on a smaller fraction of difficult samples. Diagnosis of samples that present a challenge to cytology was also improved. Samples labeled as "atypical cells," which require additional time and follow-up, were classified in high-confidence regimes in 8 of 15 cases. Further, 10 of 17 cytology-negative samples corresponding to patients with concurrent cancer were correctly classified as malignant or negative, in agreement with 6-month outcomes. This study lays the groundwork for broader validation of label-free quantitative biophysical markers for clinical diagnoses of cancer and inflammation, which could help to reduce laboratory workload and improve clinical decision-making.


Imaging stem cell distribution, growth, migration, and differentiation in 3-D scaffolds for bone tissue engineering using mesoscopic fluorescence tomography.
Regenerative medicine has emerged as an important discipline that aims to repair injury or replace damaged tissues or organs by introducing living cells or functioning tissues. Successful regenerative medicine strategies will likely depend upon a simultaneous optimization strategy for the design of biomaterials, cell-seeding methods, cell-biomaterial interactions, and molecular signaling within the engineered tissues. It remains a challenge to image three-dimensional (3-D) structures and functions of the cell-seeded scaffold in mesoscopic scale (>2 ∼ 3 mm). In this study, we utilized angled fluorescence laminar optical tomography (aFLOT), which allows depth-resolved molecular characterization of engineered tissues in 3-D to investigate cell viability, migration, and bone mineralization within bone tissue engineering scaffolds in situ.


Ethnographic perspectives on global mental health.
The field of Global Mental Health (GMH) aims to influence mental health policy and practice worldwide, with a focus on human rights and access to care. There have been important achievements, but GMH has also been the focus of scholarly controversies arising from political, cultural, and pragmatic critiques. These debates have become increasingly polarized, giving rise to a need for more dialogue and experience-near research to inform theorizing. Ethnography has much to offer in this respect. This paper frames and introduces five articles in this issue of Transcultural Psychiatry that illustrate the role of ethnographic methods in understanding the effects and implications of the field of global mental health on mental health policy and practice. The papers include ethnographies from South Africa, India, and Tonga that show the potential for ethnographic evidence to inform GMH projects. These studies provide nuanced conceptualizations of GMH's varied manifestations across different settings, the diverse ways that GMH's achievements can be evaluated, and the connections that can be drawn between locally observed experiences and wider historical, political, and social phenomena. Ethnography can provide a basis for constructive dialogue between those engaged in developing and implementing GMH interventions and those critical of some of its approaches.


Accelerated Chondrogenic Differentiation of Human Perivascular Stem Cells with NELL-1.
Osteoarthritis is the leading cause of disability in the US. Consequently, there is a pressing need for restoring the structural and functional properties of diseased articular cartilage. Yet the search for the right combination of proper target cells and growth factors for cartilage regeneration remains challenging. In this study, we first tested the intrinsic chondrogenic differentiation ability of human perivascular stem cells (hPSCs), a novel source of mesenchymal stem cells (MSCs) isolated by fluorescence-activated cell sorting (FACS) from human adipose tissue. A putative prochondrogenic growth factor, NEL-like molecule-1 (NELL-1), was added to the hPSC pellets to upregulate gene expression of chondrogenic markers, including AGGRECAN, COLLAGEN II, and COMP. Furthermore, the addition of NELL-1 to a transforming growth factor beta 3 (TGF-β3) + bone morphogenetic protein-6 (BMP-6) "cocktail" resulted in the best combinatorial stimulation in accelerating the chondrogenic differentiation of hPSCs, as evidenced by increased gene and protein expression of chondrogenic markers in a shortened induction time without elevating expression of hypertrophic, fibrotic, and osteogenic markers. Mechanistically, this acceleration rendered by NELL-1 may be partially attributed to NELL-1's upregulation of BMP receptors and TGF-β receptor type I in hPSCs for increased responsiveness to BMPs + TGF-βs. In conclusion, lipoaspirate-derived hPSCs present a novel and abundant cell source of MSCs for cartilage regeneration, and the combinatorial application of NELL-1, TGF-β3, and BMP-6 with hPSCs may remarkably enhance and accelerate cartilage repair.


Bowel disorders and its spatial trend in Manitoba, Canada.
Bowel disorders have destructive impacts on the patients social and mental aspects of life and can cause emotional distress. The risk of developing bowel incontinence also increases with age. The rate of incidence of inflammatory bowel disease in Manitoba, Canada, has been unusually raised. Therefore, it is important to identify trends in the incidence of bowel disorders that may suggest further epidemiological studies to identify risk factors and identify any changes in important factors. An important part of spatial epidemiology is cluster detection as it has the potential to identify possible risk factors associated with disease, which in turn may lead to further investigations into the nature of diseases. To test for potential disease clusters many methods have been proposed. The focused detection methods including the circular spatial scan statistic (CSS), flexible spatial scan statistic (FSS), and Bayesian disease mapping (BYM) are among the most popular disease detection procedures. A frequentist approach based on maximum likelihood estimation (MLE) has been recently used to identify potential focused clusters as well. The aforementioned approaches are studied by analyzing a dataset of bowel disorders in the province of Manitoba, Canada, from 2001 to 2010. The CSS method identified less regions than the FSS method in the south part of the province as potential clusters. The same regions were identified by the BYM and MLE methods as being potential clusters of bowel disorders with a slightly different order of significance. Most of these regions were also detected by the CSS or FSS methods. Overall, we recommend using the methods BYM and MLE for cluster detection with the similar population and structure of regions as in Manitoba. The potential clusters of bowel disorders are generally located in the southern part of the province including the eastern part of the city of Winnipeg. These results may represent real increases in bowel disorders or they may be an indication of other covariates that were not adjusted for in the model used here. Further investigation is needed to examine these findings, and also to explore the cause of these increases.


Beyond nutrient-based food indices: a data mining approach to search for a quantitative holistic index reflecting the degree of food processing and including physicochemical properties.
Processing has major impacts on both the structure and composition of food and hence on nutritional value. In particular, high consumption of ultra-processed foods (UPFs) is associated with increased risks of obesity and diabetes. Unfortunately, existing food indices only focus on food nutritional content while failing to consider either food structure or the degree of processing. The objectives of this study were thus to link non-nutrient food characteristics (texture, water activity (aw), glycemic and satiety potentials (FF), and shelf life) to the degree of processing; search for associations between these characteristics with nutritional composition; search for a holistic quantitative technological index; and determine quantitative rules for a food to be defined as UPF using data mining. Among the 280 most widely consumed foods by the elderly in France, 139 solid/semi-solid foods were selected for textural and aw measurements, and classified according to three degrees of processing. Our results showed that minimally-processed foods were less hyperglycemic, more satiating, had better nutrient profile, higher aw, shorter shelf life, lower maximum stress, and higher energy at break than UPFs. Based on 72 food variables, multivariate analyses differentiated foods according to their degree of processing. Then technological indices including food nutritional composition, aw, FF and textural parameters were tested against technological groups. Finally, a LIM score (nutrients to limit) ≥8 per 100 kcal and a number of ingredients/additives >4 are relevant, but not sufficient, rules to define UPFs. We therefore suggest that food health potential should be first defined by its degree of processing.


[Dynamics of quaternary structure of creatine kinase purified from rabbit skeletal muscles].
Using gel filtration and ultracentrifugation, the quaternary structure of muscle creatine kinase (isoenzyme MM) has been shown to depend on pH, protein concentration and ionic strength of solution. In solution the enzyme can exist not only as a dimer, but also as a monomer and tetramer with molecular weights of 40 000 and 160 000, respectively. High dilution of the protein solution and pH 6.0--5.0 provide for the monomer formation; the dimer can arise under various conditions, while the tetramer requires pH 9.0 and high protein concentration. The monomer differs from the dimer by a higher enzymatic activity, lability and needs thiol to maintain its catalytic activity. Dissociation of creatine kinase into more active subunits accounts for the increase in specific activity induced by protein dilution. It is assumed that dissociation can have physiological significance as one of the mechanisms of creatine kinase activation in muscles.


Digital image processing for pre-compensation of high-order aberrations of the human eye.
Human beings rely significantly on their visual capabilities to successfully interact with their environment. In today's technology-based world, one of the most important interaction channels is through computers. Some individuals with severe visual impairments may have difficulty in interacting with computers, even when using traditional means of visual correction (e.g., spectacles, contact lenses). This is, in part, because these correction mechanisms can only compensate for the most regular distortions or aberrations of the image in the eye. This paper proposes an image processing approach that will pre-compensate the images displayed on the computer screen, so as to counter the effect of the eye's aberrations on the image. The characterization of the eye required to perform this customized pre-compensation is the eye's Point Spread Function (PSF). The PSF can now be measured by a new generation of ophthalmic instruments generically called. Wavefront Analyzers.. The characterization provided by these instruments also includes the .higher-order aberration components. and could, therefore, lead to a more comprehensive vision correction than traditional mechanisms. The methods presented here will be explained in terms of their theoretical foundation and illustrated with results from the correction of aberrations introduced by lenses with known and constant PSFs.


Knowledge of animal appearance among sighted and blind adults.
How does first-person sensory experience contribute to knowledge? Contrary to the suppositions of early empiricist philosophers, people who are born blind know about phenomena that cannot be perceived directly, such as color and light. Exactly what is learned and how remains an open question. We compared knowledge of animal appearance across congenitally blind (n = 20) and sighted individuals (two groups, n = 20 and n = 35) using a battery of tasks, including ordering (size and height), sorting (shape, skin texture, and color), odd-one-out (shape), and feature choice (texture). On all tested dimensions apart from color, sighted and blind individuals showed substantial albeit imperfect agreement, suggesting that linguistic communication and visual perception convey partially redundant appearance information. To test the hypothesis that blind individuals learn about appearance primarily by remembering sighted people's descriptions of what they see (e.g., "elephants are gray"), we measured verbalizability of animal shape, texture, and color in the sighted. Contrary to the learn-from-description hypothesis, blind and sighted groups disagreed most about the appearance dimension that was easiest for sighted people to verbalize: color. Analysis of disagreement patterns across all tasks suggest that blind individuals infer physical features from non-appearance properties of animals such as folk taxonomy and habitat (e.g., bats are textured like mammals but shaped like birds). These findings suggest that in the absence of sensory access, structured appearance knowledge is acquired through inference from ontological kind.


Evaluation of precision medicine assessment reports of the Belgian healthcare payer to inform reimbursement decisions.
Precision medicines rely on companion diagnostics to identify patient subgroups eligible for receiving the pharmaceutical product. Until recently, the Belgian public health payer, RIZIV-INAMI, assessed precision medicines and companion diagnostics separately for reimbursement decisions. As both components are considered co-dependent technologies, their assessment should be conducted jointly from a health technology assessment (HTA) perspective. As of July 2019, a novel procedure was implemented accommodating for this joint assessment practice. The aim of this research was to formulate recommendations to improve the assessment in the novel procedure. This study evaluated the precision medicine assessment reports of RIZIV-INAMI of the last 5 years under the former assessment procedure. The HTA framework for co-dependent technologies developed by Merlin et al. for the Australian healthcare system was used as a reference standard in this evaluation. Criteria were scored as either present or not present. Thirteen assessment reports were evaluated. Varying scores between reports were obtained for the domain establishing the co-dependent relationship between diagnostic and pharmaceutical. Domains evaluating the clinical utility of the biomarker and the cost-effectiveness performed poorly, whereas the budget impact and the transfer of trial data to the local setting performed well. Based on these results we recommend three amendments for the novel procedure. (i) The implementation of the linked evidence approach when direct evidence of clinical utility is not present, (ii) incorporation of a bias assessment tool, and (iii) further specify guidelines for submission and assessment to decrease the variability of reported evidence between assessment reports.


Understanding the Digital Gap Among US Adults With Disability: Cross-Sectional Analysis of the Health Information National Trends Survey 2013.
Disabilities affect more than 1 in 5 US adults, and those with disabilities face multiple barriers in accessing health care. A digital gap, defined as the disparity caused by differences in the ability to use advanced technologies, is assumed to be prevalent among individuals with disabilities. This study examined the associations between disability and use of information technology (IT) in obtaining health information and between trust factors and IT use. We hypothesized that compared to US adults without disabilities, those with disabilities are less likely to refer to the internet for health information, more likely to refer to a health care provider to obtain health information, and less likely to use IT to exchange medical information with a provider. Additionally, we hypothesized that trust factors, such as trust toward health information source and willingness to exchange health information, are associated with IT use. The primary database was the 2013 Health Information National Trends Survey 4 Cycle 3 (N=3185). Disability status, the primary study covariate, was based on 6 questions that encompassed a wide spectrum of conditions, including impairments in mobility, cognition, independent living, vision, hearing, and self-care. Study covariates included sociodemographic factors, respondents' trust toward the internet and provider as information sources, and willingness to exchange medical information via IT with providers. Study outcomes were the use of the internet as the primary health information source, use of health care providers as the primary health information source, and use of IT to exchange medical information with providers. We conducted multivariate logistic regressions to examine the association between disability and study outcomes controlling for study covariates. Multiple imputations with fully conditional specification were used to impute missing values. We found presence of any disability was associated with decreased odds (adjusted odds ratio [AOR] 0.65, 95% CI 0.43-0.98) of obtaining health information from the internet, in particular for those with vision disability (AOR 0.27, 95% CI 0.11-0.65) and those with mobility disability (AOR 0.51, 95% CI 0.30-0.88). Compared to those without disabilities, those with disabilities were significantly more likely to consult a health care provider for health information in both actual (OR 2.21, 95% CI 1.54-3.18) and hypothetical situations (OR 1.80, 95% CI 1.24-2.60). Trust toward health information from the internet (AOR 3.62, 95% CI 2.07-6.33), and willingness to exchange via IT medical information with a provider (AOR 1.88, 95% CI 1.57-2.24) were significant predictors for seeking and exchanging such information, respectively. A potential digital gap may exist among US adults with disabilities in terms of their recent use of the internet for health information. Trust toward health information sources and willingness play an important role in people's engagement in use of the internet for health information. Future studies should focus on addressing trust factors associated with IT use and developing tools to improve access to care for those with disabilities.


Colorectal and Breast Cancer Screening Status for People in Ontario Provincial Correctional Facilities.
Primary care represents an opportunity to improve health for people who experience imprisonment, and screening for colorectal and breast cancer indicate primary care quality. The study objectives were to examine the proportion of people released from provincial correctional facilities who were overdue for colorectal or breast cancer screening on admission to the correctional facility and who were still overdue after 3 years, and to compare findings with data for the general population. Administrative data were used to identify people eligible for colorectal and breast cancer screening who were released from provincial correctional facilities in 2010 (N=3,803 and N=249, respectively) and in the general population (N=2,757,584 and N=1,099,942, respectively) in Ontario, Canada. Chi-square tests and log binomial regression models were used to compare the proportion of individuals overdue for screening on admission or on July 1, 2010 for the general population, and still overdue after 3 years. Analyses were conducted in 2017 and 2018. Compared with the general population, people in the corrections group were 1.53 times (95% CI=1.50, 1.55) more likely to be overdue for colorectal cancer screening: 77.1% (95% CI=74.3, 79.9) vs 50.5% (95% CI=50.5, 50.6), and 2.25 times (95% CI=2.06, 2.46) more likely to be overdue for breast cancer screening: 65.9% (95% CI=56.2, 76.8) vs 29.3% (95% CI=29.2, 29.4, both p<0.001). They were also more likely to still be overdue 3 years later: 62.6% (95% CI=60.1, 65.2) vs 33.6% (95% CI=33.5, 33.6) for colorectal cancer and 52.2% (95% CI=43.6, 62.0) vs 20.2% (95% CI=20.1, 20.3) for breast cancer (both p<0.001). People who experience imprisonment are less likely than the general population to access colorectal and breast cancer screening. This suggests the need to strengthen primary care for this population. Specifically, efforts should be made to improve access to colorectal and breast cancer screening, through health promotion, program delivery, and linkage to community services while in correctional facilities.


Computational survey of peptides derived from disulphide-bonded protein loops that may serve as mediators of protein-protein interactions.
Bioactive cyclic peptides derived from natural sources are well studied, particularly those derived from non-ribosomal synthetases in fungi or bacteria. Ribosomally synthesised bioactive disulphide-bonded loops represent a large, naturally enriched library of potential bioactive compounds, worthy of systematic investigation. We examined the distribution of short cyclic loops on the surface of a large number of proteins, especially membrane or extracellular proteins. Available three-dimensional structures highlighted a number of disulphide-bonded loops responsible for the majority of the likely binding interactions in a variety of protein complexes, due to their location at protein-protein interfaces. We find that disulphide-bonded loops at protein-protein interfaces may, but do not necessarily, show biological activity independent of their parent protein. Examining the conservation of short disulphide bonded loops in proteins, we find a small but significant increase in conservation inside these loops compared to surrounding residues. We identify a subset of these loops that exhibit a high relative conservation, particularly among peptide hormones. We conclude that short disulphide-bonded loops are found in a wide variety of biological interactions. They may retain biological activity outside their parent proteins. Such structurally independent peptides may be useful as biologically active templates for the development of novel modulators of protein-protein interactions.


Duration of serum antibody response to rabies vaccination in horses.
OBJECTIVE To investigate the impact of age and inferred prior vaccination history on the persistence of vaccine-induced antibody against rabies in horses. DESIGN Serologic response evaluation. ANIMALS 48 horses with an undocumented vaccination history. PROCEDURES Horses were vaccinated against rabies once. Blood samples were collected prior to vaccination, 3 to 7 weeks after vaccination, and at 6-month intervals for 2 to 3 years. Serum rabies virus-neutralizing antibody (RVNA) values were measured. An RVNA value of ≥ 0.5 U/mL was used to define a predicted protective immune response on the basis of World Health Organization recommendations for humans. Values were compared between horses < 20 and ≥ 20 years of age and between horses inferred to have been previously vaccinated and those inferred to be immunologically naïve. RESULTS A protective RVNA value (≥ 0.5 U/mL) was maintained for 2 to 3 years in horses inferred to have been previously vaccinated on the basis of prevaccination RVNA values. No significant difference was evident in response to rabies vaccination or duration of protective RVNA values between horses < 20 and ≥ 20 years of age. Seven horses were poor responders to vaccination. Significant differences were identified between horses inferred to have been previously vaccinated and horses inferred to be naïve prior to the study. CONCLUSIONS AND CLINICAL RELEVANCE A rabies vaccination interval > 1 year may be appropriate for previously vaccinated horses but not for horses vaccinated only once. Additional research is required to confirm this finding and characterize the optimal primary dose series for rabies vaccination.


Mass psychogenic illness after vaccination.
When vaccines are administered to groups, the physical reactions of the recipients may be similar, causing a form of mass reaction, the mechanism for which is the same as that for mass reactions from other causes. These phenomena have been categorised as mass psychogenic illness (MPI), and have been defined as the collective occurrence of a constellation of symptoms suggestive of organic illness but without an identified cause in a group of people with shared beliefs about the cause of the symptom(s). A review of the literature shows that such outbreaks have been reported in differing cultural and environmental settings including developing and industrialised countries, in the work place, on public transport, in schools, and the military. The perceived threats have been against agents such as food poisoning, fire and toxic gases. Whatever the place or perceived threat, the response seems to be similar. The symptoms generally included headache, dizziness, weakness, and loss of consciousness. Once under way, MPIs are not easy to stop. Incidents reported in the literature show that they can quickly gather momentum and can be amplified by the press who disseminate information rapidly, escalating the events. Management of such mass events can be extremely difficult. Should the public health official in charge continue to try and determine the cause, or should this person call off the entire investigation? It is suggested here that once vaccines are identified as a probable cause of the phenomenon, a dismissive approach may actually be harmful. Unless the spokesperson has already earned a high level of trust, the public are not likely to be convinced easily that nothing was wrong with the vaccine until it has been tested. An increased awareness of MPIs on the part of organisers of future mass vaccination campaigns seems appropriate. Immunisation managers should be aware that mass immunisation campaigns could generate such mass reactions. It is therefore essential that surveillance/reporting systems for reporting adverse events be improved before such campaigns. A mass campaign using a smallpox vaccine should be accompanied by a surveillance system capable of distinguishing between multiple cases of conventionally understood vaccine reactions and outbreaks of mass psychogenic illness.


Epidemic management: a Caribbean university's response to the 2009 H1N1 outbreak.
In June 2009, the World Health Organization (WHO) declared a global influenza pandemic (1). During the throes of the epidemic, the Caribbean region recorded 22 deaths and 350 hospitalizations. Jamaica reported most of the deaths in the region--7, followed by Trinidad and Tobago--5 and Barbados--3; the remaining 7 deaths occurred in other Caribbean countries (2). In June, Jamaica reported its first case (3) and some three months later in September 2009, The University of the West Indies (UWI) Mona Campus experienced an outbreak of the 2009 H1N1 virus infection. As one of the three main Campuses of the leading tertiary institution in the English-speaking Caribbean (others being in Trinidad and Tobago and Barbados in addition to an 'Open Campus' which serves the non-Campus territories), the UWI Mona Campus serves some 15 000 students. These students originate mainly from the Caribbean; North American and African students also comprise the student population. The University Health Centre provides primary care services to a community of students, staff (3000) and their dependents, retirees, visiting staff and students.


Unveiling hole trapping and surface dynamics of NiO nanoparticles.
The research effort in mesoporous p-type semiconductors is increasing due to their potential application in photoelectrochemical energy conversion devices. In this paper an electron-hole pair is created by band-gap excitation of NiO nanoparticles and the dynamics of the electron and the hole is followed until their recombination. By spectroscopic characterization it was found that surface Ni3+ states work as traps for both electrons and holes. The trapped electron was assigned to a Ni2+ state and the trapped hole to a "Ni4+" state positioned close to the valence band edge. The recombination kinetics of these traps was studied and related with the concept of hole relaxation suggested before. The time scale of the hole relaxation was found to be in the order of tens of ns. Finally the spectroscopic evidence of this relaxation is presented in a sensitized film.


Interproximal biofilm removal by intervallic use of a sonic toothbrush compared to an oral irrigation system.
The purpose of this in-vitro study was to investigate the potential of biofilm removal in interproximal tooth regions using intervallic cleaning with an oral irrigator or a sonic toothbrush. Three-species biofilms (Streptococcus mutans (OMZ 918), Streptococcus oralis SK 248 (OMZ 60), Actinomyces naeslundii (OMZ 745)) were grown on hydroxyapatite discs for 3 days in culture media. Every 24 h, specimens were incubated for 15 min in resazurin solution (i.e., culture medium and 10 % v/v alamarBlue®) to measure the metabolic activity with a fluorescence spectrophotometer in relative fluorescence units (rfu) at baseline. Then, specimens were fixed in interproximal holding devices and underwent treatment with an oral irrigator (WF; Waterpik® Sensonic WP-100E), an active sonic toothbrush (WPa), or an inactive sonic toothbrush (WPi; Waterpik® Sensonic SR-3000E) for 10 s (n = 18/group). Untreated biofilms served as controls (CO). After treatment, bacterial activity was re-measured, and specimens were re-grown in fresh medium for 24 h until next cleaning procedure. Altogether, cleaning was repeated in intervals of three treatment days (d1, d2, d3). After d3, SEM images were taken (n = 8) and CFU was measured (n = 3). Metabolic activity was analyzed for each disc separately, rfu values were averaged for d1 to compare initial biofilm stability, and ratios of baseline and post-treatment values were compared. Results were analyzed using ANOVA with the post-hoc Scheffé test, or Kruskal-Wallis with post-hoc Mann-Whitney test. Median baseline rfu-values of d1 resulted in 7821.8 rfu (interquartile range = 5114.5). Highest reduction in metabolic activity was recorded significantly for the oral irrigator used for 10 s (residual activity per day d1: WF 17.9 %, WPa 58.8 %, WPi 82.5 %, CO 89.6 %; d2: WF 36.8 %, WPa 85.2 %, WPi 82.5 %, CO 90.0 %; d3: WF 17.2.%, WPa 79.6 %, WPi 96.3 %, CO 116.3 %). SEM images of untreated specimens (CO) and specimens treated with the sonic toothbrush (WPa and WPi) showed huge amounts of biofilm, while oral irrigator-treated specimens (WF) revealed barely any bacteria. CFU data confirmed the graduations between the groups. Cleaning of interproximal regions achieved better success with an oral irrigator as compared to the use of a sonic toothbrush. (350/350 words).


Cocaine metabolism accelerated by a re-engineered human butyrylcholinesterase.
Plasma butyrylcholinesterase (BChE) is important in the metabolism of cocaine, but natural human BChE has limited therapeutic potential for detoxication because of low catalytic efficiency with cocaine. Here we report pharmacokinetics of cocaine in rats treated with A328W/Y332A BChE, an excellent cocaine hydrolase designed with the aid of molecular modeling. Compared with wild-type BChE, this enzyme hydrolyzes cocaine with 40-fold improved k(cat) (154 min(-1) versus 4.1 min(-1)) and only slightly increased K(M) (18 microM versus 4.5 microM). In rats given this hydrolase (3 mg/kg i.v.) 10 min before cocaine challenge (6.8 mg/kg i.v.), cocaine half-life was reduced from 52 min to 18 min. Mirroring the reductions of plasma cocaine were large increases in benzoic acid, a product of BChE-mediated cocaine hydrolysis. All other pharmacokinetic parameters confirmed a large, dose-dependent acceleration of cocaine removal by the injected cocaine hydrolase. These results show that A328W/Y332A, an efficient cocaine hydrolase in vivo as well as in vitro, might promote cocaine detoxication in a clinical setting.


Killers on the loose: Immunotherapeutic strategies to improve NK cell-based therapy for cancer treatment.
Natural killer (NK) cells are innate lymphocytes that control tumor progression by not only directly killing cancer cells, but also by regulating other immune cells, helping to orchestrate a coordinated anti-tumor response. However, despite the tremendous potential that this cell type has, the clinical results obtained from diverse NK cell-based immunotherapeutic strategies have been, until recent years, rather modest. The intrinsic regulatory mechanisms that are involved in the control of their activation as well as the multiple mechanisms that tumor cells have developed to escape NK cell-mediated cytotoxicity likely account for the unsatisfactory clinical outcomes. The current approaches to improve long-term NK cell function are centered on modulating different molecules involved in both the activation and inhibition of NK cells, and the latest data seems to advocate for combining strategies that target multiple aspects of NK cell regulation. In this review, we summarize the different strategies (such as engineered NK cells, CAR-NK, NK cell immune engagers) that are currently being used to take advantage of this potent and complex immune cell.


Health system preparedness in noncommunicable diseases: Findings from two states Odisha and Kerala in India.
"Non-communicable diseases (NCDs) are the raging reality of today's world and have moved up the priority list of most countries worldwide including India. The government of India has launched programs such as National Programme for Prevention and Control of Cancer, Diabetes, Cardiovascular disease, and Stroke, but little is known about preparedness of health system to address NCDs. A qualitative study was conducted in the states of Odisha and Kerala to document the government stakeholder perspectives of health system preparedness for NCDs. The study revealed that both the states have a NCD preparedness structure but need strengthening at various levels. The present human resource is inadequate, over-burdened, and requires specific skills. The lack of workforce adversely affects the service delivery mechanism. The study highlighted that there is a gap in the timely release of funds and also lack of evidence-based application if the information that is made available. There is a clear need to fill the gaps and strengthen the identified areas at various levels to address the increasing NCD burden. There is also a need to more in-depth and large-scale studies on such issues which act as external monitoring mechanisms thereby assisting the policy makers and program managers in with relevant and scientific evidence to bring reforms in the health system.


Provider Discussion about Lifestyle by Cancer History: A Nationally Representative Survey.
Providers are uniquely positioned to encourage health-promoting behaviors, particularly among cancer survivors where patients develop trust in providers. We utilized the National Health Interview Survey to identify adults who reported a visit to a provider in the prior year (44,385 individuals with no cancer history and 4,792 cancer survivors), and reported prevalence of provider discussions on weight loss, physical activity, diet, and smoking. We used generalized linear mixed models to examine predicted prevalence of provider lifestyle discussions by cancer history overall, and among those who do not meet body mass index (BMI), activity, or smoking guidelines. Among those with a BMI of 25-<60 kg/m2, 9.2% of those with a cancer history and 11.6% of those without a cancer history reported being told to participate in a weight loss program (P < 0.001). Overall, 31.7% of cancer survivors and 35.3% of those with no cancer history were told to increase their physical activity (P < 0.001). Only 27.6% of cancer survivors and 32.2% of those with no cancer history reported having a general discussion of diet (P < 0.001). Among smokers, 67.3% of cancer survivors and 69.9% of those with no cancer history reported counseling on smoking (P = 0.309). Fewer cancer survivors, who are at increased risk for health complications, are reporting provider discussions about critical lifestyle issues than those with no cancer history. Our nationally representative results suggest that providers are missing an opportunity for influencing patient lifestyle factors, which could lead to mitigation of late and long-term effects of treatment.


Interaction between the BAG1S isoform and HSP70 mediates the stability of anti-apoptotic proteins and the survival of osteosarcoma cells expressing oncogenic MYC.
The oncoprotein MYC has the dual capacity to drive cell cycle progression or induce apoptosis, depending on the cellular context. BAG1 was previously identified as a transcriptional target of MYC that functions as a critical determinant of this cell fate decision. The BAG1 protein is expressed as multiple isoforms, each having an array of distinct biochemical functions; however, the specific effector function of BAG1 that directs MYC-dependent cell survival has not been defined. In our studies the human osteosarcoma line U2OS expressing a conditional MYC-ER allele was used to induce oncogenic levels of MYC. We interrogated MYC-driven survival processes by modifying BAG1 protein expression. The function of the separate BAG1 isoforms was investigated by depleting cells of endogenous BAG1 and reintroducing the distinct isoforms. Flow cytometry and immunoblot assays were performed to analyze the effect of specific BAG1 isoforms on MYC-dependent apoptosis. These experiments were repeated to determine the role of the HSP70 chaperone complex in BAG1 survival processes. Finally, a proteomic approach was used to identify a set of specific pro-survival proteins controlled by the HSP70/BAG1 complex. Loss of BAG1 resulted in robust MYC-induced apoptosis. Expression of the larger isoforms of BAG1, BAG1L and BAG1M, were insufficient to rescue survival in cells with oncogenic levels of MYC. Alternatively, reintroduction of BAG1S significantly reduced the level of apoptosis. Manipulation of the BAG1S interaction with HSP70 revealed that BAG1S provides its pro-survival function by serving as a cofactor for the HSP70 chaperone complex. Via a proteomic approach we identified and classified a set of pro-survival proteins controlled by this HSP70/BAG1 chaperone complex that contribute to the BAG1 anti-apoptotic phenotype. The small isoform of BAG1, BAG1S, in cooperation with the HSP70 chaperone complex, selectively mediates cell survival in MYC overexpressing tumor cells. We identified a set of specific pro-survival clients controlled by the HSP70/BAG1S chaperone complex. These clients define new nodes that could be therapeutically targeted to disrupt the survival of tumor cells driven by MYC activation. With MYC overexpression occurring in most human cancers, this introduces new strategies for cancer treatment.


Naturally secreted amyloid-beta increases mammalian target of rapamycin (mTOR) activity via a PRAS40-mediated mechanism.
Reducing the mammalian target of rapamycin (mTOR) activity increases lifespan and health span in a variety of organisms. Alterations in protein homeostasis and mTOR activity and signaling have been reported in several neurodegenerative disorders, including Alzheimer disease (AD); however, the causes of such deregulations remain elusive. Here, we show that mTOR activity and signaling are increased in cell lines stably transfected with mutant amyloid precursor protein (APP) and in brains of 3xTg-AD mice, an animal model of AD. In addition, we show that in the 3xTg-AD mice, mTOR activity can be reduced to wild type levels by genetically preventing Aβ accumulation. Similarly, intrahippocampal injections of an anti-Aβ antibody reduced Aβ levels and normalized mTOR activity, indicating that high Aβ levels are necessary for mTOR hyperactivity in 3xTg-AD mice. We also show that the intrahippocampal injection of naturally secreted Aβ is sufficient to increase mTOR signaling in the brains of wild type mice. The mechanism behind the Aβ-induced mTOR hyperactivity is mediated by the proline-rich Akt substrate 40 (PRAS40) as we show that the activation of PRAS40 plays a key role in the Aβ-induced mTOR hyperactivity. Taken together, our data show that Aβ accumulation, which has been suggested to be the culprit of AD pathogenesis, causes mTOR hyperactivity by regulating PRAS40 phosphorylation. These data further indicate that the mTOR pathway is one of the pathways by which Aβ exerts its toxicity and further support the idea that reducing mTOR signaling in AD may be a valid therapeutic approach.


Unfolding and extraction of a transmembrane alpha-helical peptide: dynamic force spectroscopy and molecular dynamics simulations.
An atomic force microscope (AFM) was used to visualize CWALP(19)23 peptides ((+)H(3)N-ACAGAWWLALALALALALALWWA-COO(-)) inserted in gel-phase DPPC and DSPC bilayers. The peptides assemble in stable linear structures and domains. A model for the organization of the peptides is given from AFM images and a 20 ns molecular dynamics (MD) simulation. Gold-coated AFM cantilevers were used to extract single peptides from the bilayer through covalent bonding to the cystein residue. Experimental and simulated force curves show two distinct force maxima. In the simulations these two maxima correspond to the extraction of the two pairs of tryptophan residues from the membrane. Unfolding of the peptide precedes extraction of the second distal set of tryptophans. To probe the energies involved, AFM force curves were obtained from 10 to 10(4) nm/s and MD force curves were simulated with 10(8)-10(11) nm/s pulling velocities (V). The velocity relationship with the force, F, was fitted to two fluctuation adhesive potential models. The first assumes the pulling produces a constant bias in the potential and predicts an F approximately ln (V) relationship. The second takes into account the ramped bias that the linker feels as it is being driven out of the adhesion complex and scales as F approximately (ln V)2/3.


Evaluation of a one-piece ceramic implant used for single-tooth replacement and three-unit fixed partial dentures: a prospective cohort clinical trial.
The aim of this clinical trial was to evaluate the safety and efficiency of a one-piece zirconia oral implant after 1 year of function. Two centers included 60 subjects in need of implant-supported single-tooth restorations or three-unit bridges. A total of 71 zirconia one-piece implants were placed and immediately restored with a temporary reconstruction for at least 2 months. The final veneered zirconia restorations were then cemented and followed for 6 months and 1 year after insertion of the restorations. At each visit, a clinical evaluation was performed to analyze biological parameters of the implants and the neighboring teeth. A standardized periapical radiograph was taken at implant insertion, at the placement of the restorations and at the 1-year follow-up. Sixty patients with 71 implants (48 in the mandible, 23 in the maxilla) were included in this study and provided with 11 bridges and 49 crowns. Two patients with three implants (one bridge and one single crown) could not be evaluated. One patient lost his implant 5 weeks after implant insertion. Based on 58 patients, the mean survival rate was 98.3% after one year when the implants of the two patients that did not show up were not counted as lost. The mean marginal bone loss from implant insertion to the 1-year follow-up after the final prosthetic restoration was 0.78 mm with a standard deviation of 0.79 mm. The probing depth around the implants increased from 2.7 mm at insertion of the prosthetic reconstruction to 3.5 mm one year after insertion. The probing depth around the adjacent teeth remained stable at 2.5 mm. At the 1-year recall, the difference was significant. The clinical attachment levels at implants and teeth were not different at the 1-year follow-up with 3.1 mm at tooth and implant sites. The presently tested one-piece ceramic implant was successful in replacing single tooth and three-unit gaps after one year of function. Further long-term data are necessary to verify these initial findings.


Enabling the democratization of the genomics revolution with a fully integrated web-based bioinformatics platform.
Continued advancements in sequencing technologies have fueled the development of new sequencing applications and promise to flood current databases with raw data. A number of factors prevent the seamless and easy use of these data, including the breadth of project goals, the wide array of tools that individually perform fractions of any given analysis, the large number of associated software/hardware dependencies, and the detailed expertise required to perform these analyses. To address these issues, we have developed an intuitive web-based environment with a wide assortment of integrated and cutting-edge bioinformatics tools in pre-configured workflows. These workflows, coupled with the ease of use of the environment, provide even novice next-generation sequencing users with the ability to perform many complex analyses with only a few mouse clicks and, within the context of the same environment, to visualize and further interrogate their results. This bioinformatics platform is an initial attempt at Empowering the Development of Genomics Expertise (EDGE) in a wide range of applications for microbial research.


Translating Pediatric Hospital Interpreters' Feedback From Difficult Conversations into Improved Communication.
Background: Medical interpreters are critical mediators in communication with pediatric subjects and families to include participation in difficult conversations. Objective: The objective of this pilot study was to provide suggestions from medical interpreters to palliative care teams as to how to effectively incorporate medical interpreters into end-of-life conversations. Methods: Participants included pediatric hospital-based medical interpreters who had interpreted for at least 1 end-of-life conversation in the pediatric hospital setting. A total of 11 surveys were completed by medical interpreters. The study consisted of a written 12-item survey with a follow-up focus group to further explore survey themes. Results: The translation of cultural contexts, awareness of the mixed messages the family received from health care teams, and the emotional intensity of the interactions were depicted as the most challenging aspects of the medical interpreter's role. Despite these challenges, 9 interpreters reported they would willingly be assigned for interpreting "bad news" conversations if given the opportunity (82%). Medical interpreters recognized their relationship with the family and their helping role for the family as meaningful aspects of interpreting even in difficult conversations. Medical interpreters shared 7 thematic suggestions for improved communication in language-discordant visits: content review, message clarity, advocacy role, cultural understanding, communication dynamics, professionalism, and emotional support. Conclusions: As experts in cultural dynamics and message transmission, the insights of medical interpreters can improve communication with families.


Evolution of the literature identifying physicians' roles in leadership, clinical development, and practice of the subspecialty of emergency medical services.
The 2007 Institute of Medicine report entitled Emergency Medical Services at the Crossroads identified a need for the establishment of physician subspecialty certification in emergency medical services (EMS). The purpose of this study was to identify and explore the evolution of publications that define the role of the physician in EMS systems in the United States. Three comprehensive searches were undertaken to identify articles that define the physician's role in the leadership, clinical development, and practice of EMS. Independent reviewers then evaluated these articles to further determine whether the articles identified the physician's role in EMS. Then, identified articles were classified by the type of publication in order to evaluate the transition from a non-peer reviewed to peer-reviewed literature base and an analysis was performed on the differences in the growth between these two groups. In addition, for the peer-reviewed articles, an analysis was performed to identify the proportion of articles that were quantitative versus qualitative in nature. The comprehensive review identified 1,504 articles. Ninety articles were excluded due to lack of relevance to the US. The remaining 1,414 articles were reviewed, and 194 papers that address the physician's role within EMS systems were identified; 72 additional articles were identified by hand search of references for a total of 266 articles. The percentage of peer-reviewed articles has increased steadily over the past three decades. In addition, the percentage of quantitative articles increased from the first decade to the second and third decades. This comprehensive review demonstrates that over the past 30 years an evidence base addressing the role of the physician in EMS has developed. This evidence base has steadily evolved to include a greater proportion of peer-reviewed, quantitative literature.


Serum 'uracil plus uridine' levels in normal subjects and their possible significance.
A microbiological method for the assay of uracil is described. The growth of the test organism is supported by uracil and also by uridine but not by uridylic acid. The method therefore measures uracil and uridine together. The ;uracil + uridine' level, expressed as uracil, has been measured in blood from 144 normal subjects ranging in age from cord blood to the eighth decade. The mean level of 22 mu mol/l (0.25 mg%) in cord blood decreases to 15 mumol/l (0.17 mg%) in adults over the age of 20. There is no difference between the sexes. Uracil is of interest because (a) it is a constituent base of RNA, (b) it is the precursor of two of the bases thymine and cytosine that enter into the composition of DNA, and (c) under certain circumstances it has mutagenic properties. The last is dependent upon the existence of two tautomeric forms of uracil, the common keto form which pairs normally with adenine and the rare enol form which pairs with guanine. A mistake in base pairing which allows uracil in its enol form to enter the DNA molecule and pair with guanine can result in a G = C --> A = T base transition in the DNA molecule. The molecular mechanism involved as well as the possible bearing on somatic mutation are discussed.


Cigarette smoke induces molecular responses in respiratory tissues of ApoE(-/-) mice that are progressively deactivated upon cessation.
Cigarette smoking is the primary etiology of chronic obstructive pulmonary disease (COPD) and a risk factor for both lung and cardiovascular (CV) diseases, which are rarely investigated concomitantly. Although smoking cessation shows clear CV risk benefit, lung-related disease risk remains higher in former smokers than in never smokers. We sought to determine the differential molecular responses of murine respiratory tissues to better understand the toxicity pathways involved in smoking-related disease risk and those related to the benefits of smoking cessation. ApoE(-/-) mice were exposed to mainstream cigarette smoke (CS) or a smoking cessation-mimicking protocol for up to 6 months and transcriptomics analysis of nasal epithelium and lung parenchyma performed. We supported our gene expression profiling approach with standard lung histopathology and bronchoalveolar lavage fluid (BALF) analysis. Many BALF analytes involved in functions ranging from inflammation to cell proliferation and tissue remodeling were found elevated in BALF. Gene expression levels of these molecules were also increased in lung tissue, suggesting that the inflammatory response was the result of local tissue activation and the contribution of recruited inflammatory cells. Gene set enrichment analysis (GSEA) of expression data from murine lungs and nasal epithelium showed distinct activation patterns of inflammation, complement, and xenobiotic metabolism pathways during CS exposure that were deactivated upon smoking cessation. Pathways involved in cell proliferation and tissue remodeling were activated by CS and progressively deactivated upon smoke exposure cessation. Differential CS-mediated responses of pulmonary and nasal tissues reflect common mechanisms but also the varying degrees of epithelial functional specialization and exposure along the respiratory tract.


Photographic response to x-ray irradiation. I: Estimation of the photographic error statistic and development of analytic density-intensity equations.
Formulations for specular optical density as a function of incident x-ray intensity are shown to be inadequate, theoretically and compared with available data. Approximations assuming low intensities, grain densities, or energies yield significant error in typical emulsions. Unjustifiable simplifications limit analysis and consequent results. The avoidance of assumptions leads to models for rough and smooth emulsion surfaces, which correspond to Kodak 101-01 and DEF-392 emulsion types. The self-consistent use of spherical grains yields scaling that is dependent on emulsion roughness. We obtained improvement over standard formulations, avoiding the empirical character of earlier models and associated parameterization. The correlation of grain locations and occluded emulsion area is approximated within monolayer depths but neglected between layers. Effects of the incident angle from a broad source, scattering, and photoelectrons are considered. The models presented herein apply to the vacuum UV and x-ray energies from 9 eV to 20 keV and may be preferred over alternative models at lower energies, densities greater than unity, emulsions with high grain fractions, or where interpolation over energy ranges is desired. Error contributions may be dominated by intensity statistics or densitometry statistics. Both are inadequate in medium-density regimes. We have derived estimates including pseudo-binomial grain development statistics, using a summation over layers.


Spontaneous Resolution of Symptomatic Thoracic Spine Calcified Disc Herniation: A Case Report and Literature Review.
Calcification of intervertebral disc (IVD) is an uncommon but well-defined clinical syndrome in children. Despite its benign nature, occasionally, calcified nucleus pulposus can herniate into the spinal canal. We report an interesting case of calcified disc herniation in the thoracic spine. Although it resulted in cord compression and syrinx formation, it well responded to conservative management. An 8-year-old girl presented with back pain and lower limbs hyperreflexia. The spinal imaging revealed that calcification within IVD, associated with huge calcified disc herniation on the T5/6, resulted in cord compression and syrinx formation within the spinal cord from T4/5 through T9/10. Despite cord compression and syrinx formation, the patient underwent conservative management. After a 6-month follow-up, the control imaging showed complete resolution of the calcified disc herniation and decreased syrinx size. In the patients with a calcified herniated IVD, in the absence of motor neurological deficit, conservative management consisting of bed rest, lifestyle adjustment, weight loss, and brace or collar wearing combined with a close follow-up can result in spontaneous regression of calcification, improvement of symptoms, and partial to complete resolution of the underlying pathology.


Quantitative studies of the sensitivity of normal and neoplastic mouse cells to the cytotoxic action of isoantibodies.
Experiments were performed to study the mechanisms of the sensitivity of normal and neoplastic mouse cells to the cytotoxic action of humoral isoantibodies. A quantitative in vitro absorption procedure was used to compare the residua] hemagglutinin titer of a certain isoantiserum after the absorption of a known volume with known cell numbers. The cell numbers were adjusted so as to yield the same average total cell surface area. Lymph node cells and various solid and ascites tumor cells of strain A and C3H origin were used. It was found that different cell types differed with regard to their concentration of isoantigenic surface receptors. High correlation coefficients were found between the cytotoxic sensitivity and the concentration of isoantigenic receptors. It was shown that tumor cells which were partially or completely resistant to a single isoantiserum could be made sensitive to humoral isoantibodies provided that they were treated with mixtures of antisera, produced in different donor genotypes against the same target genotype, since each individual genotype reacts against a different antigen combination. This supports the concept that cytotoxic sensitivity is related to the surface concentration of reactive sites. Hematopoietic liver cells of newborn mice, which are completely resistant to isoantibodies, and spleen and lymph node cells of adult animals which are highly sensitive, were compared with regard to their ability to take up fluorescein-conjugated normal mouse serum in vitro. No differences were found between their visible pinocytotic uptake. Similar experiments were performed with various ascites tumor cells having different cytotoxic sensitivity. All tumor cells studied had a low pinocytosing ability which was not correlated to their sensitivity to humoral isoantibodies.


Racecadotril for the treatment of severe acute watery diarrhoea in children admitted to a tertiary hospital in Kenya.
Diarrhoea is the second most common cause of death in children under 5 years of age in Kenya. It is usually treated with oral rehydration, zinc and continued feeding. Racecadotril has been in use for over 2 decades; however, there is a paucity of data regarding its efficacy from Africa. The objectives of this study were: to compare the number of stools in the first 48 hours in children with severe gastroenteritis requiring admission and treated with either racecadotril or placebo, to study the impact of racecadotril on duration of inpatient stay as well as duration of diarrhoea and to describe the side effect profile of racecadotril. This was a randomised, double-blinded, placebo-controlled trial. It enrolled children between the age of 3 and 60 months who were admitted with severe acute gastroenteritis. They received either racecadotril or placebo in addition to oral rehydration solution (ORS) and zinc and were followed up daily. 120 children were enrolled into the study. There were no differences in the demographics or outcomes between the 2 groups. Stools at 48 hours: median (IQR) of 5 (3-7) and 5 (2.5-7.5), respectively; p=0.63. The duration of inpatient stay: median (IQR): 4 days (1.5-6.5) and 4.5 (1.8-6.3); p=0.71. The duration of illness: 3 days (2-4) and 2 days (1-3); p=0.77. The relative risk of a severe adverse event was 3-fold higher in the drug group but was not statistically significant (95% CI 0.63 to 14.7); p=0.16. Racecadotril has no impact on the number of stools at 48 hours, the duration of hospital stay or the duration of diarrhoea in children admitted with severe gastroenteritis and managed with ORS and zinc. PACTR201403000694398; Pre-results.


IL-4 expression at the onset of islet inflammation predicts nondestructive insulitis in nonobese diabetic mice.
In nonobese diabetic mice, autoimmune diabetes progresses in an age-linked and gender-dependent manner. Insulitis begins in male and female mice at approximately 1 mo of age; however, 70 to 90% of females, but only 10 to 20% of males, become diabetic by 6 mo. Multiple studies propose that proinflammatory Th1 and immunomodulatory Th2 cytokines impact diabetes pathogenesis, but the role of these cytokines in spontaneous diabetes progression is not yet clear. We used quantitative reverse-transcriptase-coupled PCR to analyze expression of cytokines and APC costimulatory molecules in the islets of 20- to 180-day-old male and female nonobese diabetic littermates, and identified three stages in diabetes progression. At 1 to 2 mo of age, islet-infiltrating T cells displayed a Th1 cytokine bias in females, and a Th2 cytokine bias in males. In females, stage II (2-3 mo of age) was characterized by an increase in islet-infiltrating T cells, APC, and Th1 cytokines, whereas male infiltrates did not increase in size, and Th1 cytokine expression continued to decline during this interval. Islet infiltration reached a plateau (stage III) in 3- to 4-mo-old females, months before overt diabetes onset. Our data imply that Th cytokine expression in early insulitis exerts substantial impact on beta cell destruction and overt diabetes. A clinical implication of our results is that young individuals in the early stages of insulitis are ideal candidates for therapeutic intervention to minimize beta cell destruction and morbidity.


A brief review of higher dietary protein diets in weight loss: a focus on athletes.
Thermodynamics dictates that for body weight (i.e. stored substrate) loss to occur a person must ingest less energy than they expend. Athletes, who owing to their oftentimes large daily energy expenditures, may have greater flexibility than non-athletes in this regard; however, they may also have different goals for weight loss. In particular, weight lost may be less important to an athlete than from which compartment the weight is lost: fat or lean. A critical question is thus, what balance of macronutrients might promote a greater fat loss, a relative retention of lean mass, and still allow athletic performance to remain uncompromised? It is the central thesis of this review that dietary protein should be a nutrient around which changes in macronutrient composition should be framed. The requirement for protein to sustain lean mass increases while in negative energy balance and protein, as macronutrient, may have advantages with respect to satiety during energy balance, and it may allow greater fat loss during a negative energy balance. However, athletes should be mindful of the fact that increasing dietary protein intake while in negative energy balance would come at the 'expense' of another macronutrient. Most recently there has been interest in lower carbohydrate diets, which may not allow performance to be sustained given the importance of dietary carbohydrate in high-intensity exercise. The relative merits of higher protein diets for athletes are discussed.


Global conformation of the Escherichia coli replication factor DnaC protein in absence and presence of nucleotide cofactors.
Global conformational and oligomeric states of the Escherichia coli replicative factor DnaC protein in the absence and presence of magnesium and nucleotide cofactors, ATP and ADP, and their fluorescent analogues, MANT-ATP and MANT-ADP, have been examined using analytical sedimentation velocity and time-dependent fluorescence anisotropy techniques. In solution, the DnaC protein exists exclusively as a monomer over a large protein concentration range. The value of s(degrees) (20, w)= 2.45 +/- 0.07 S indicates that the protein molecule has an elongated shape. When modeled as a prolate ellipsoid of revolution, the hydrated DnaC protein has an axial ratio of 4.0 +/- 0.6 with long axis a = 112 A and the short axis b = 28 A, respectively. The presence of magnesium or nucleotide cofactors, ATP or ADP, does not affect the global conformation of the protein and its monomeric state. These data indicate that recently found cooperative interactions between the DnaC molecules, in the complex with the DnaB helicase, are induced by the binding to the helicase, i.e., they are not the intrinsic property of the DnaC protein. Fluorescence anisotropy decays of the DnaC-MANT-ATP and DnaC-MANT-ADP complexes indicate that the protein has a rigid global structure on the nanosecond time scale, little affected by the nucleotide cofactors. Nevertheless, the complex with ATP has a more flexible structure, while the complex with ADP is more rigid, with the protein molecule assuming a more elongated shape. Magnesium exerts control only on the complex with the ATP analogue. In the absence of magnesium, the ATP analogue is firmly held in the binding site. In the presence of Mg(2+), this fixed location is released and the analogue is allowed to assume a flexible conformational state. The significance of the results for the functioning of the DnaC protein is discussed.


Measurement of longitudinal displacement using lateral shearing cyclic path optical configuration setup and phase shifting interferometry.
We present a technique for the measurement of longitudinal displacement using a lateral shearing cyclic path optical configuration (CPOC) setup and phase shifting interferometry. In the technique, a plane mirror mounted on a linear translation stage, placed slightly away from the focal plane of a lens, introduces a longitudinal focal shift to the incident focusing beam. The resulting spherical wavefront emerging from the lens is sheared into two orthogonally polarized beams using the CPOC setup. By applying polarization phase shifting interferometry (PPSI), the longitudinal focal shift of the beam focus is calculated by determining the slope of the optical path difference variation between the sheared beams. Similarly, the additional focal shift introduced due to longitudinal translation of the mirror, by an unknown amount, is determined using PPSI. Half of the difference between the two longitudinal focal shifts measured gives the longitudinal displacement of the mirror. The technique can be used for an extended range of distance measurement. The novelty of the technique is the introduction of CPOC for the distance measurement. The advantages of the technique compared to other related methods are discussed.


Gold-198 Wires Used To Study Movements of Small Mammals.
Eastern harvest mice, Reithrodontomys humulis humulis (Audubon and Bachman), were tagged with subcutaneously inserted 20 gauge gold-198 wires 10 mm long and varying in activity from 0.7 to 4.5 mc. None of the tags inserted by this method were lost or had any apparent effect on the animals. The movements of the tagged mice were successfully traced with a portable Geiger counter.


Automated detection of malignant features in confocal microscopy on superficial spreading melanoma versus nevi.
In-vivo reflectance confocal microscopy (RCM) shows promise for the early detection of superficial spreading melanoma (SSM). RCM of SSM shows pagetoid melanocytes (PMs) in the epidermis and disarray at the dermal-epidermal junction (DEJ), which are automatically quantified with a computer algorithm that locates depth of the most superficial pigmented surface [D(SPS)(x,y)] containing PMs in the epidermis and pigmented basal cells near the DEJ. The algorithm uses 200 noninvasive confocal optical sections that image the superficial 200 μm of ten skin sites: five unequivocal SSMs and five nevi. The pattern recognition algorithm automatically identifies PMs in all five SSMs and finds none in the nevi. A large mean gradient ψ (roughness) between laterally adjacent points on D(SPS)(x,y) identifies DEJ disruption in SSM ψ = 11.7 ± 3.7 [-] for n = 5 SSMs versus a small ψ = 5.5 ± 1.0 [-] for n = 5 nevi (significance, p = 0.0035). Quantitative endpoint metrics for malignant characteristics make digital RCM data an attractive diagnostic asset for pathologists, augmenting studies thus far, which have relied largely on visual assessment.


First Report of Botryosphaeria iberica Associated with Dieback and Tree Mortality of Monterey Cypress (Cupressus macrocarpa) in Algeria.
Stem cankers and branches showing bark discoloration, fissuring, resin exudation leading to dieback, crown wilting, and tree mortality have been observed since late spring 2008 on 40-year-old Cupressus macrocarpa (Hartw.) trees planted in forests mixed with Juniperus oxycedrus L. and Acer monspessulanum L. in Taffet, near Ain Abbessa, in the district of Bougaa, Algeria (36°18'57″N; 05°06'33″E; 1,400 m elevation). In 2010, approximately 60% of the C. macrocarpa trees were diseased. For fungal isolations, cankered branches were surface sterilized with ethanol. After removal of the outer bark, fragments of necrotic inner bark taken from the margin of cankers were plated on potato dextrose agar (PDA). Most of the colonies were identified as Botryosphaeria iberica (Phillips, Luque & Alves) based on comparison of morphological traits and DNA sequences with known isolates of the fungus (1). Pestalotiopsis funerea colonies were also obtained, although with less frequency. B. iberica colonies on PDA were dark green with aerial mycelium and optimum growth at 25°C. Pycnidia were produced after 3 weeks of incubation at 20°C under a 12-h near UV light photoperiod on water agar amended with autoclaved cypress seeds. Conidia were brown, one-septate, oval to oblong, and 24.2 (20.1 to 27.4) × 11.2 μm (8.8 to 14.1) (n= 50). An isolate was deposited at the Centralbureau voor Schimmelculture as CBS 130984. DNA was extracted from freeze-dried mycelium and amplified using primers ITS1 and ITS4. The amplified DNA sequence of B. iberica isolate CBS 130984 from Algeria (GenBank Accession No. JN836991) showed 100% homology with sequences of B. iberica isolates obtained from dead and cankered bark of oaks from Spain and Italy (GenBank Accession Nos. AY573216, AY573214, AY573213, AY573210, AY573202, and AY573201). Stem inoculations were performed in the greenhouse on 10 4-year-old, grafted plants of C. macrocarpa growing in 5-liter pots using isolate CBS 130984. A 3-mm plug taken from the margin of a colony grown on PDA for 1 week was inserted in a circular wound of the same size made in the bark with a cork borer where the stem diameter was approximately 1 cm. Inoculations were repeated in June 2010 and June 2011. Five months after inoculations, small rounded to elongated lesions (1.0 to 2.5 cm long), sometimes with resin exuding cracks, were visible on all inoculated stems. Control trees, inoculated with sterile PDA plugs, showed no canker development. B. iberica was successfully reisolated from the necrotic bark surrounding the inoculation sites. No significant differences in canker size were observed between the two replicated experiments. Some Botryosphaeria species that are found on a variety of hosts are also known to cause cankers and dieback of cypress; among these are B. stewensii, B. obtusa, B. dothidea, and B. ribis, often acting as weak pathogens (2,3). Considered weakly virulent in causing dieback of grapevine (4) and, to our knowledge, reported here for the first time on Cupressaceae, B. iberica caused cankers and dieback of C. macrocarpa trees that had probably been weakened by repeated drought events occurring in Algeria during the last 10 years. References: (1) A. Phillips et al. Mycologia 97:513, 2005. (2) E. Punithalingam and J. M. Waller. IMI Descriptions of Fungi and Bacteria 40, Sheet 394, 1973; (3) E. Punithalingam and P. Holliday. IMI Descriptions of Fungi and Bacteria. 40, Sheet 395, 1973; (4) R. Úrbez-Torres et al. Plant Dis. 93:584, 2009.


Economic impact of the liver condemnation of cattle infected with Fasciola hepatica in the Peruvian Andes.
Fasciolosis is a zoonotic food- and waterborne infection related to great economic losses on ruminant production. In order to determine the prevalence of bovine liver condemnations due to fasciolosis in an abattoir at the Central Andes of Peru, we evaluated the records of an authorized slaughterhouse in Huancayo, at 3300 m above sea level, over 16 months. Subsequently, the average weights of condemned viscera and carcasses were also estimated to determine the direct and indirect economic losses. Fasciolosis was the main cause of liver condemnation, with a prevalence of 55.72% (3926/7046; CI 95%: 56.88-54.56%). The livers infected were more than double the amount of livers supplied to markets in this period. Although the average price of liver in the Peruvian Andes is very cheap, approximately 16.45 tons of livers were destroyed, estimating a direct economic loss of US$ 35,080 (CI 95%: US$ 29,178-40,938). The indirect loss due to decreased body weight gain (calculated using 10% of the average weight of a healthy animal carcass) was estimated at US$ 354,570 (CI 95%: US$ 387,908-US$ 321,227). Fasciolosis is an important zoonotic disease that threatens food security in the Andean region, with major repercussions on the income of the livestock industry and the availability of cheap protein sources.


Nutrilyzer: a tool for deciphering atomic stoichiometry of differentially expressed paralogous proteins.
Organisms try to maintain homeostasis by balanced uptake of nutrients from their environment. From an atomic perspective this means that, for example, carbon:nitrogen:sulfur ratios are kept within given limits. Upon limitation of, for example, sulfur, its acquisition is triggered. For yeast it was shown that transporters and enzymes involved in sulfur uptake are encoded as paralogous genes that express different isoforms. Sulfur deprivation leads to up-regulation of isoforms that are poor in sulfur-containing amino acids, that is, methinone and cysteine. Accordingly, sulfur-rich isoforms are down-regulated. We developed a web-based software, doped Nutrilyzer, that extracts paralogous protein coding sequences from an annotated genome sequence and evaluates their atomic composition. When fed with gene-expression data for nutrient limited and normal conditions, Nutrilyzer provides a list of genes that are significantly differently expressed and simultaneously contain significantly different amounts of the limited nutrient in their atomic composition. Its intended use is in the field of ecological stoichiometry. Nutrilyzer is available at http://nutrilyzer.hs-mittweida.de. Here we describe the work flow and results with an example from a whole-genome Arabidopsis thaliana gene-expression analysis upon oxygen deprivation. 43 paralogs distributed over 37 homology clusters were found to be significantly differently expressed while containing significantly different amounts of oxygen.


Correcting base-assignment errors in repeat regions of shotgun assembly.
Accurate base-assignment in repeat regions of a whole genome shotgun assembly is an unsolved problem. Since reads in repeat regions cannot be easily attributed to a unique location in the genome, current assemblers may place these reads arbitrarily. As a result, the base-assignment error rate in repeats is likely to be much higher than that in the rest of the genome. We developed an iterative algorithm, EULER-AIR, that is able to correct base-assignment errors in finished genome sequences in public databases. The Wolbachia genome is among the best finished genomes. Using this genome project as an example, we demonstrated that EULER-AIR can 1) discover and correct base-assignment errors, 2) provide accurate read assignments, 3) utilize finishing reads for accurate base-assignment, and 4) provide guidance for designing finishing experiments. In the genome of Wolbachia, EULER-AIR found 16 positions with ambiguous base-assignment and two positions with erroneous bases. Besides Wolbachia, many other genome sequencing projects have significantly fewer finishing reads and, hence, are likely to contain more base-assignment errors in repeats. We demonstrate that EULER-AIR is a software tool that can be used to find and correct base-assignment errors in a genome assembly project.


Current trends in inflammatory and immunomodulatory mediators in sepsis.
Sepsis refers to severe systemic inflammation in response to invading pathogens. An overwhelming immune response, as mediated by the release of various inflammatory mediators, can lead to shock, multiple organ damage, and even death. Cytokines, proteases, lipid mediators, gaseous substances, vasoactive peptides, and cell stress markers play key roles in sepsis pathophysiology. Various adhesion molecules and chemokines sequester and activate neutrophils into the target organs, further augmenting inflammation and tissue damage. Although the anti-inflammatory substances counterbalance proinflammatory mediators, prolonged immune modulation may cause host susceptibility to concurrent infections, thus reflecting enormous challenge toward developing effective clinical therapy against sepsis. To understand the complex interplay between pro- and anti-inflammatory phenomenon in sepsis, there is still an unmet need to study newly characterized mediators. In addition, revealing the current trends of novel mediators will upgrade our understanding on their signal transduction, cross-talk, and synergistic and immunomodulating roles during sepsis. This review highlights the latest discoveries of the mediators in sepsis linking to innate and adaptive immune systems, which may lead to resolution of many unexplored queries.