Preoperative serum prostate specific antigen does not reflect biochemical failure rates after radical prostatectomy in men with large volume cancers.
We compared pathological findings with prostate specific antigen (PSA) failure rates following radical prostatectomy for large volume cancers (6 cc or greater). A total of 191 men whose radical prostatectomy specimen had a cancer volume of 6 cc or greater were followed for a mean of 3.6 years (range 0.3 to 11.1) and 112 (58.6%) had PSA failure (PSA 0.07 ng./ml. or greater and increasing). Percent Gleason grade 4/5 (the Stanford modified Gleason scale), cancer volume, seminal vesicle invasion, regional lymph nodes, capsular penetration, positive surgical margin, location of the largest cancer in the peripheral or transition zone, prostate weight, patient age, preoperative PSA and clinical stage were analyzed using univariate and multivariate Cox proportional hazards analyses. In univariate regression analysis percent Gleason grade 4/5, lymph node involvement, cancer volume, cancer location in the peripheral zone, capsular penetration and positive surgical margins were significant predictors of biochemical failure. Seminal vesicle invasion, preoperative serum PSA, patient age, prostate weight and clinical stage were not statistically significant. Forward stepwise, multivariate analysis showed that percent Gleason grade 4/5 (p <0.0001, relative risk ratio 2.498), cancer location in the peripheral zone (p = 0.0097, 1.887), cancer volume (p = 0.0157, 1.691) and lymph node involvement (p = 0.0317, 1. 666) were the only independent predictors of biochemical failure. When 52 men with organ confined, large volume prostate cancer were analyzed separately, univariate and multivariate analyses showed that only cancer location in the peripheral zone (p = 0.0021, relative risk ratio 13.473) and percent Gleason grade 4/5 (p = 0. 0449, 4.111) were independent predictors of failure. Percent Gleason grade 4/5, cancer location in the peripheral zone, cancer volume and lymph node involvement have prognostic value in large volume prostate cancer. Cancer location in the peripheral zone and percent Gleason grade 4/5 are the most powerful predictors of biochemical failure in men whose cancer is 6 cc or greater and contained in the prostatic capsule. Preoperative serum PSA is not helpful in distinguishing biochemical failure rates in these large volume cancers whether they are organ confined or not.


The effect of L3T4 T cell depletion on the pathogenesis of Theiler's murine encephalomyelitis virus infection in CBA mice.
Theiler's murine encephalomyelitis virus (TMEV) gives rise to a biphasic disease of the central nervous system (CNS) following intracranial inoculation of susceptible strains of mice. The early phase, during the first month, resembles poliomyelitis and in the late phase the mice suffer from inflammatory demyelination reminiscent of multiple sclerosis. In order to investigate the role of helper T cells in the acute and chronic phases of the disease we depleted mice of their L3T4 T cells in vivo with rat monoclonal antibodies, prior to infection and prior to the onset of clinical signs of demyelination. Mice depleted of their helper cells failed to produce antibodies to TMEV and consequently were unable to clear virus from the CNS and died within the first month of infection. Depletion of T cells before the demyelinating phase of the disease resulted in a marked decrease in the incidence of disease from 77% of the immunocompetent animals with clinical signs of paralysis to 36%. Immunocompetent TMEV-infected mice also developed antibodies against myelin suggesting that autoimmune mechanisms may play a role in TMEV-induced demyelination.


Permeability characteristics of human endothelial monolayers seeded on different extracellular matrix proteins.
To investigate whether endothelial monolayer permeability changes induced by inflammatory mediators are affected by the extracellular matrix protein used for cell seeding. Human umbilical venular endothelial cells (HUVEC) were grown to confluent monolayers on membranes coated with either collagen, fibronectin or gelatin. The permeability to albumin and dextran was then assessed, both under normal conditions and after treatment with tumor necrosis factor-alpha (TNF-alpha) and bacterial lipopolysaccharide (LPS). With any of the three protein coatings, tight junctions were formed all over the monolayers. The permeability of the coated membranes to albumin and dextran was reduced strongly by confluent monolayers; the relative reduction was similar for the three matrix proteins used. Pre-incubation of the monolayers with either TNF-alpha or LPS increased permeability dose dependently. However, the relative increase due to either treatment was independent of the protein used for membrane coating. The extracellular matrix protein used for initial seeding of endothelial cultures plays a minor role in determining the permeability changes induced in HUVEC monolayers by inflammatory mediators.


Return to work and occupational physicians' management of common mental health problems--process evaluation of a randomized controlled trial.
The aim of this study was to examine the adherence of occupational physicians (OP) to the Dutch guideline on the management of common mental health problems and its effect on return to work as part of the process evaluation of a trial comparing adherence to the guideline to care as usual. The first hypothesis was that guideline adherence among the "guideline group" will be higher compared to the "usual care group". The second hypothesis was that better guideline adherence by the occupational physician will be associated with earlier return to work. In a randomized controlled trial, five participating OP had to provide care based on the Dutch guideline to 240 police workers with common mental health problems (the "guideline group"). The same OP had to provide usual care to the participants in the control group (the "usual care" group), including minimal involvement and easy access to a psychologist. In evaluating the process, we assessed guideline adherence via an audit of medical files, using 20 guideline-based performance indicators. Mean rates of guideline adherence were related to the duration until first and full return to work, using a Cox proportional hazards model. The mean rate of the sum score of guideline adherence was 10 in a range of 0-20 [standard deviation (SD) 1.8] and did not significantly differ between the intervention and control group. Mean better guideline adherence showed a statistically significant association with a shorter time to first and full return to work [hazard ratio 1.1; 95% confidence interval (95% CI) 1.0-1.2], which was explained by keeping more regular contact with the worker and the work system and better monitoring of stagnation or return to work. No contrast in guideline adherence was found between guideline-based versus usual care. This can be explained by contamination between the guideline and usual care group. Even though guideline adherence was only average, better adherence predicted earlier return to work. Guidelines for management of common mental health problems and return to work should focus on regular contact with the worker and the work organisation.


A Trauma Center-Based Study for Victims of Domestic Violence: Protecting Women of Richmond and Surrounding Counties.
Domestic violence (DV) is a major health issue on both a national and global scale. In Richmond County, Georgia, there are 18.1 calls per 1000 women for DV, exceeding the national average of 5.9 and Georgia average of 15.3 calls per 1000 women. The goal of the study is to map epidemiologic and spatial trends in communities of DV survivors thereby assessing the feasibility of a prospective intervention initiative for high-risk areas. In partnership with "SafeHomes", a local women's shelter, a retrospective review of physical addresses was compiled from first-time residential clients 18 and older from January 1, 2019 to December 31, 2019. Hot zones were overlaid with Census tracts to assess sociodemographic correlates including race, ethnicity, age, poverty, education, and employment status. From all records (n = 85), 5 hot zones were identified. Analysis of census tract data revealed minority predominance in all hot zones, in addition to an unemployment rate above the state average (23%, 12%, 8%, 5%, and 19%, respectively, vs 3.4%). DV affects both the inner city and suburban areas of Richmond County. Hot zone frequency was disproportionately increased in the urban center where community demographics showed minority predominance, especially of African-Americans. These hot zones also have more individuals living below the federal poverty line and experience unemployment rates greater than the state and national average of 3.4 and 3.5, respectively. This study demonstrated that mapping domestic violence epidemiologic and spatial trends is possible, allowing for targeted support and intervention to reduce the rate of domestic violence.


Rescue of Na+ affinity in aspartate 928 mutants of Na+,K+-ATPase by secondary mutation of glutamate 314.
The Na(+),K(+)-ATPase binds Na(+) at three transport sites denoted I, II, and III, of which site III is Na(+)-specific and suggested to be the first occupied in the cooperative binding process activating phosphorylation from ATP. Here we demonstrate that the asparagine substitution of the aspartate associated with site III found in patients with rapid-onset dystonia parkinsonism or alternating hemiplegia of childhood causes a dramatic reduction of Na(+) affinity in the α1-, α2-, and α3-isoforms of Na(+),K(+)-ATPase, whereas other substitutions of this aspartate are much less disruptive. This is likely due to interference by the amide function of the asparagine side chain with Na(+)-coordinating residues in site III. Remarkably, the Na(+) affinity of site III aspartate to asparagine and alanine mutants is rescued by second-site mutation of a glutamate in the extracellular part of the fourth transmembrane helix, distant to site III. This gain-of-function mutation works without recovery of the lost cooperativity and selectivity of Na(+) binding and does not affect the E1-E2 conformational equilibrium or the maximum phosphorylation rate. Hence, the rescue of Na(+) affinity is likely intrinsic to the Na(+) binding pocket, and the underlying mechanism could be a tightening of Na(+) binding at Na(+) site II, possibly via movement of transmembrane helix four. The second-site mutation also improves Na(+),K(+) pump function in intact cells. Rescue of Na(+) affinity and Na(+) and K(+) transport by second-site mutation is unique in the history of Na(+),K(+)-ATPase and points to new possibilities for treatment of neurological patients carrying Na(+),K(+)-ATPase mutations.


Mathematical Discrepancies of the Tono-Pen Applanation Tonometer.
The purpose of the study was to determine if Tono-Pen tonometers use simple average and coefficient of variation (CV) algorithms to calculate intraocular pressure (IOP). IOPs were measured as part of routine ocular examination in 152 client-owned dogs. Using 11 Tono-Pen's, a total of 778 averaged readings were collected. Individual IOP readings, and average IOP and CV displayed by the instrument, were recorded. Average IOP and CV were then manually calculated from individual readings and compared with those displayed by the instrument. The mean absolute difference between the calculated and displayed average IOP was 1.37±2.01 mm Hg (P<0.001). In 6% of cases, the calculated average IOP was 5 to 15 mm Hg different from the displayed average IOP. The difference between the displayed and calculated average IOP was significantly higher in hypertensive eyes with displayed IOP≥25 mm Hg. Calculated CV was equal to, lower than, or greater than displayed CV in 28.6%, 1.5%, and 69.7% of cases, respectively. In 17.6% of cases, calculated CV was >20%, but displayed CV was <5%. Receiver operating characteristic analysis could not correlate number of individual IOP readings with magnitude of difference in average IOP. Calculated average IOP and CV differ significantly from values displayed by the instrument, especially at higher IOPs. A difference of ≥5 mm Hg between calculated and displayed average IOP seen in 6% of cases may impact clinical judgement. Displayed CV<5% does not correlate with accurate IOP measurement based on individual results.


Altered functional connectivity of primary visual cortex in adults with strabismus and amblyopia: a resting-state fMRI study.
Functional connectivity of the primary visual cortex was explored with resting functional magnetic resonance imaging among adults with strabismus and amblyopia and healthy controls. We used the two-sample test and receiver operating characteristic curves to investigate the differences in mean functional connectivity values between the groups with strabismus and amblyopia and healthy controls. Compared with healthy controls, functional connectivity values in the left Brodmann areas 17, including bilateral lingual/angular gyri, were reduced in groups with strabismus and amblyopia. Moreover, functional connectivity values in the right Brodmann area 17, including left cuneus, right inferior occipital gyrus, and left inferior parietal lobule, were reduced in adults with strabismus and amblyopia. Our findings indicate that functional connectivity abnormalities exist between the primary visual cortex and other regions. This may be the basis of the pathological mechanism of visual dysfunction and stereovision disorders in adults with strabismus and amblyopia.


Using Twitter to increase content dissemination and control educational content with Presenter Initiated and Generated Live Educational Tweets (PIGLETs).
Live-tweeting during educational presentations is typically learner-generated and can lead to misquoted information. Presenter curated tweets have not been well described. We created Presenter Initiated and Generated Live Educational Tweets (PIGLETs) with the goal to broaden the reach of educational conferences. We hypothesized that using PIGLETs would increase the reach and exposure of our material. We developed a prospective single-arm intervention study performed during the "Not Another Boring Lecture" workshops presented at two national conferences in 2015. Presenters tweeted PIGLETs linked to unique hashtags #NotAnotherBoringLecture and #InnovateMedEd. Analytic software was used to measure the following outcomes: (1) number of tweets published by presenters versus learners, (2) reach (users exposed to content containing the hashtag), and (3) exposure (total number of times content was delivered). One hundred and twenty-six participants attended the workshops. A total of 636 tweets (including retweets) were sent by presenters containing the study hashtags, compared with 162 sent by learners. #NotAnotherBoringLecture reached 47,200 users and generated 136,400 impressions; #InnovateMedEd reached 36,400 users and generated 79,100 impressions. PIGLETs allowed presenters to reach a significant number of learners, as well as control the content delivered through Twitter. PIGLETs can be used to augment educational sessions beyond the physical confines of the classroom.


Cost-effectiveness of BTK inhibitors vs bendamustine and rituximab in chronic lymphocytic leukemia patients.
Background: We aimed to compare the cost-effectiveness of bruton tyrosine kinase inhibors (BTKis) versus bendamustine-rituximab (R-bendamustine) as a first-line treatment for Chinese patients with relapsed or refractory chronic lymphocytic leukemia. Methods: A partitioned survival model was constructed using TreeAge Pro 2022 software and transition probabilities were estimated from the reported survival probabilities using parametric survival modeling. One-way analysis and probabilistic sensitivity analysis were performed to explore the uncertainty of the modeling results. In addition, several scenario analyses were evaluated. Results: In comparison to R-bendamustine, zanubrutinib had an incremental cost-effectiveness ratio (ICER; life years) and ICER (quality-adjusted life years) of US$12,173.38 and $17,983.40, respectively. While ibrutinib had a higher ICER relative to R-bendamustine. Conclusion: Zanubrutinib was cost-effective for patients with relapsed or refractory chronic lymphocytic leukemia in China.


Childhood leukaemia in areas with different radon levels: a spatial and temporal analysis using GIS.
To evaluate the relation between exposure to ground radon levels and leukaemia among children using existing population and disease registers. Ecological correlation study. The county of Ostergötland in Sweden. Every child born in the county between 1979 and 1992 was mapped to the property centroid coordinates by linking addresses in the population and property registers. Population maps were overlaid with radon maps and exposure at birth and each subsequent year was quantified as high, normal, low or unknown. This was analysed with data from the tumour registry. Standardised mortality ratios (SMRs) were calculated using the age and sex specific rates for Sweden for the year 1995. 90 malignancies occurred among 53 146 children (498 887 person years) who formed the study population. SMRs for acute lymphatic leukaemia (ALL) among children born in high, normal and low risk areas were 1.43, 1.17 and 0.25 respectively. The relative risk for the normal risk group and high risk group as compared with the low risk group was 4.64 (95% CI 1.29, 28.26) and 5. 67 (95% CI 1.06, 42.27). The association between ALL and continued residence at normal or high risk areas showed a similar trend. No association between radon risk levels and any other malignancy was seen. Children born in and staying at areas where the risk from ground radon has been classified as low are less likely to develop ALL than those born in areas classified as normal and high risk.


Spatial pattern analysis of deforestation in the northeast of Minas Gerais State, Brazil.
Understanding the spatial pattern of a particular geographic phenomenon such as deforestation is a key issue to establish monitoring programs to prevent the depletion of natural resources. Thus, the goal of this study was to assess the spatial pattern of deforested areas in the Pardo and Jequitinhonha River basins using Ripley's K function. First, we mapped all deforested areas in these basins using Landsat multispectral imagery from 2007 to 2015. Then, we used the Ripley's K function to test for spatial interactions between deforestation events. Our results showed that deforestations predominantly occur in a clustering spatial pattern in these basins. Spatial statistical analyses as Ripley's K function may provide a baseline for deforestation monitoring, as well as allowing us to understand the spatial pattern of deforestation in different natural ecosystems, especially in countries like Brazil, where the territorial dimension presents a great difficulty for the effectiveness of deforestation monitoring.


Thermally induced fluid reversed hexagonal (H(II)) mesophase.
In the present study we characterized the microstructures of the Lc and HII phases in a glycerol monooleate (GMO)/tricaprylin (TAG)/water mixture as a function of temperature. We studied the factors that govern the formation of a low-viscosity HII phase at relatively elevated temperatures (>35 degrees C). This phase has very valuable physical characteristics and properties. The techniques used were differential scanning calorimetry (DSC), wide- and small-angle X-ray scattering (WAXS and SAXS, respectively), NMR (self-diffusion and (2)H NMR), and Fourier transform infrared (FTIR) spectroscopies. The reverse hexagonal phase exhibited relatively rapid flow of water in the inner channels within the densely packed cylindrical aggregates of GMO with TAG molecules located in the interstices. The existence of two water diffusion peaks reflects the existence of both mobile water and hydration water at the GMO-water interface (hydrogen exchange between the GMO hydroxyls and water molecules). Above 35 degrees C, the sample became fluid yet hexagonal symmetry was maintained. The fluidity of the HII phase is explained by a significant reduction in the domain size and also perhaps cylinder length. This phenomenon was characterized by higher mobility of the GMO, lower mobility of the water, and a significant dehydration process.


Ethnicity and race influence the folate status response to controlled folate intakes in young women.
Population-based studies report differences in folate status indicators among Mexican American (MA), African American (AA) and Caucasian (CA) women. It is unclear, however, whether these differences are due to variations in dietary folate intake. The present study was designed to investigate the influence of ethnicity/race on folate status parameters in MA, AA, and CA women (18-45 y; n = 14 in each group) under conditions of strictly controlled folate intake. In addition, the adequacy of the 1998 folate U.S. recommended dietary allowance (RDA), 400 micro g/d as dietary folate equivalents (DFE), for non-Caucasian women was assessed. Subjects (n = 42) with the methylenetetrahydrofolate reductase 677 CC genotype consumed a low-folate diet (135 micro g DFE/d) for 7 wk followed by repletion with 400 (7 MA, 7 AA, 7 CA) or 800 micro g DFE/d (7 MA, 7 AA, 7 CA) for 7 wk. AA women had lower (P </= 0.05) blood folate concentrations and excreted less (P </= 0.05) urinary folate throughout folate depletion and repletion with 400 and/or 800 micro g DFE/d compared with MA and/or CA women. MA women had lower (P </= 0.05) plasma total homocysteine (tHcy) throughout folate depletion and during repletion with 400 micro g DFE/d relative to the other ethnic/racial groups. Repletion with the 1998 folate U.S. RDA led to normal blood folate and plasma tHcy for all 3 ethnic/racial groups. Collectively, these data demonstrate that ethnicity/race is an important determinant of folate status under conditions of strictly controlled dietary folate intake and support the adequacy of the 1998 folate U.S. RDA for the 3 largest ethnic/racial groups in the United States.


Lipid bodies: cytoplasmic organelles important to arachidonate metabolism in macrophages and mast cells.
Much has been learned about the biochemical nature and pharmacologic activity of the products of arachidonic acid (AA) oxidation, but relatively little is known about the structures in nucleated cells into which AA is incorporated and from which it is initially mobilized. To address this question, we administered 3H-AA or other 3H-fatty acids in vitro to human lung mast cells and alveolar macrophages as well as to mouse and guinea pig peritoneal macrophages. The subcellular distribution of 3H label was assessed by electron microscopic autoradiography, and the nature of cell-associated 3H-lipids was determined by thin layer chromatography. Autoradiographic analysis of human lung mast cells localized virtually all of the 3H-AA to cytoplasmic lipid bodies. Lipid bodies are roughly spherical, variable osmiophilic, nonmembrane-bound structures that appear in the cytoplasm of a wide variety of cells, but we have found that these lipid bodies occur with increased frequency in granulocytes, macrophages, and mast cells at sites of inflammatory, immunologic, or neoplastic processes. Macrophages also incorporated 3H-AA predominantly into cytoplasmic lipid bodies. In contrast to mast cells, however, macrophages incorporated 3H-AA into the plasma membrane as well. Stimulation of macrophage phagocytosis resulted in striking alterations of the relationships of lipid bodies to intracellular membranes, so that many lipid bodies appeared adjacent to phagolysosomes. In addition, some phagolysosomes contained 3H label, which along with other morphologic evidence suggested that lipid bodies may discharge their contents into these structures. Mast cell and macrophage cytoplasmic lipid bodies appear to represent a major site of intracellular storage and metabolism of products of AA and perhaps other fatty acids taken up from the external milieu. These heretofore neglected organelles may thus influence cellular function in a wide variety of adaptive or pathologic processes.


Optimization and Evaluation of Site-Identification by Ligand Competitive Saturation (SILCS) as a Tool for Target-Based Ligand Optimization.
Chemical fragment cosolvent sampling techniques have become a versatile tool in ligand-protein binding prediction. Site-identification by ligand competitive saturation (SILCS) is one such method that maps the distribution of chemical fragments on a protein as free energy fields called FragMaps. Ligands are then simulated via Monte Carlo techniques in the field of the FragMaps (SILCS-MC) to predict their binding conformations and relative affinities for the target protein. Application of SILCS-MC using a number of different scoring schemes and MC sampling protocols against multiple protein targets was undertaken to evaluate and optimize the predictive capability of the method. Seven protein targets and 551 ligands with broad chemical variability were used to evaluate and optimize the model to maximize Pearson's correlation coefficient, Pearlman's predictive index, correct relative binding affinity, and root-mean-square error versus the absolute experimental binding affinities. Across the protein-ligand sets, the relative affinities of the ligands were predicted correctly an average of 69% of the time for the highest overall SILCS protocol. Training the FragMap weighting factors using a Bayesian machine learning (ML) algorithm led to an increase to an average 75% relative correct affinity predictions. Furthermore, once the optimal protocol is identified for a specific protein-ligand system average predictabilities of 76% are achieved. The ML algorithm is successful with small training sets of data (30 or more compounds) due to the use of physically correct FragMap weights as priors. Notably, the 76% correct relative prediction rate is similar to or better than free energy perturbation methods that are significantly computationally more expensive than SILCS. The results further support the utility of SILCS as a powerful and computationally accessible tool to support lead optimization and development in drug discovery.


[The making of madness: counterculture and anti-psychiatry].
The 1950s and especially the 1960s saw constant revisions of social values and customs, with young people's movements playing a major role, above all the so-called counter-culture. The powers-that-be categorized the behavior and attitudes of the movement's followers as constituting madness. This making of madness gave rise to a stream of thought known as anti-psychiatry, which calls into question the very essence of psychiatry. The present article criticizes the psychiatric models of that era and draws links between counter-culture movements and anti-psychiatry.


Design and characterization of an injectable pericardial matrix gel: a potentially autologous scaffold for cardiac tissue engineering.
Following ischemic injury in the heart, little to no repair occurs, causing a progressive degeneration of cardiac function that leads to congestive heart failure. Cardiac tissue engineering strategies have focused on designing a variety of injectable scaffolds that range in composition from single-component materials to complex extracellular matrix (ECM)-derived materials. In this study, the pericardial ECM, a commonly used biomaterial, was investigated for use as an injectable scaffold for cardiac repair. It was determined that a solubilized form of decellularized porcine pericardium could be injected and induced to gel in vivo, prompting investigation with human pericardium, which has the decided advantage of offering an autologous therapy. Characterization showed that the matrix gels retained components of the native pericardial ECM, with extant protein and glycosaminoglycan content identified. The results of an in vitro migration assay indicate that the porcine pericardial matrix is a stronger chemoattractant for relevant cell types, but in vivo results showed that the two materials caused statistically similar amounts of neovascularization, demonstrating feasibility as injectable treatments. Potential stem cell mobilization was supported by the presence of c-Kit+ cells within the matrix injection regions. With this work, the pericardium is identified as a novel source for an autologous scaffold for treating myocardial infarction.


Simultaneous analytical optimization of variational parameters in Gaussian-type functions with full configuration interaction of multicomponent molecular orbital method by elimination of translational and rotational motions: application to isotopomers of the hydrogen molecule.
We have extended the multicomponent molecular orbital (MCMO) method to the full-configuration interaction (full-CI) fully variational molecular orbital method by elimination of translational and rotational motion components from total Hamiltonian. In the MCMO scheme, the quantum effects of protons and deuterons as well as electrons can be directly taken into account. All variational parameters in the full-CI scheme, i.e., exponents and centers (alpha and R) in the Gaussian-type function (GTF) basis set as well as the CI coefficients, are simultaneously optimized by using their analytical gradients. The total energy of the H(2) molecule calculated using the electronic [6s3p2d1f] and nuclear [1s1p1d1f] GTFs is -1.161 726 hartree, which can be compared to the energy of -1.164 025 hartree reported using a 512 term-explicitly correlated GTF calculation. Although the d- and f-type nuclear GTFs contribute to the improvement of energy convergence, the convergence of electron-nucleus correlation energy is slower than that of electron-electron one. The nuclear wave functions are delocalized due to the electron-nucleus correlation effect compared to the result of Hartree-Fock level of MCMO method. In addition, the average internuclear distances of all diatomic molecules are within 0.001 A of the previously reported experimental results. The dipole moment of the HD molecule estimated by our method is 8.4 x 10(-4) D, which is in excellent agreement with the experimental result of (8-10) x 10(-4) D.


Mechanical circulatory support in children: bridge to transplant versus recovery.
The number of children and adolescents suffering from heart failure is increasing dramatically. Some of these patients will progress to need advanced therapies in the form of mechanical circulatory support (MCS). Over the past few years, increased attention has been focused on clinical use of existing devices as well the development of pediatric-specific ventricular assist devices (VADs). As in adult populations, these devices offer unique opportunities to successfully support children as a bridge-to-transplant, but increasing data suggest that bridge-to-recovery and bridge-to-destination are also viable options in select pediatric populations. Herein, we will review existing approaches as well describe future potential MCS options.


[Forgotten hepatitis: the hepatitis E].
Hepatitis E is caused by the Hepatitis E Virus (HEV), entericaly transmitted. In areas with poor sanitation, HEV is responsible for high endemicity and major outbreaks. In countries with high sanitary conditions, HEV is a zoonosis responsible for sporadic cases, and is encountered in travellers returning from endemic areas. The clinical manifestations are not distinguishable from that caused by other causes of acute viral hepatitis. Fulminant hepatitis are encountered in 1 to 4% of acute hepatitis E. Remarquably, mortality among pregnant women is very high. Diagnosis can be made by serological testing and eventually viral detection by PCR. HEV can cause chronic infection in immunocompromised patients. There is no specific therapy. There is no vaccine currently available.


Chiral metal nanoparticle-catalyzed asymmetric C-C bond formation reactions.
Chiral ligand-modified metal nanoparticles possess an attractive potential for application in asymmetric synthesis. This article focuses on chiral-nanoparticle-catalyzed asymmetric C-C bond formation reactions and discusses the nature of the active species.


Therapeutic exercise in the treatment of patients with lumbar spinal stenosis.
Lumbar spinal stenosis is a condition that may cause significant pain and associated disability, especially in older patients. It is being recognized with increasing frequency as the population continues to age, and is the most common diagnosis associated with lumbar spine surgery in patients older than 65 years of age. The natural history of lumbar spinal stenosis is not necessarily one of progressive deterioration. Conservative treatment is advocated in patients with mild to moderate symptoms of lumbar spinal stenosis, and may include therapeutic exercise. The therapeutic exercise program must be prescribed with a thorough understanding of the contributing pathoanatomic and pathophysiologic factors, and should be tailored to each patient based on his or her history and physical examination. Components of the program are described in detail and include specific stretching and strengthening exercises, general conditioning exercises, and education in proper posture and body mechanics. Randomized controlled studies are needed to help clarify the indications for conservative versus surgical treatment, to determine which components of the therapeutic exercise program are the most beneficial, and to compare outcomes after conservative or surgical measures.


Recurrence of neuroblastoma following prolonged remission.
A 6-year-old girl with unresectable and metastatic neuroblastoma had a complete remission with irradiation to the primary tumor and systemic administration of cyclophosphamide. The patient was disease-free for 16 years but had an explosive recurrence of tumor six weeks after a hysterectomy. Although no clear cause-and-effect relationship exists between the surgery and the recurrence, this case illustrates that a recurrence is possible after a long disease-free interval. Diagnostic and therapeutic procedures in such patients should be undertaken with caution.


The concept of quality of life of dying persons in the context of health care.
Considerable research has addressed quality of life and quality of care; however, it has not addressed adequately the experiences of patients and their families at the end of life. Health care may have a greater effect on quality of life during the dying process than it normally does. Building on research and expert review, a conceptual framework is presented that specifies and integrates quality of life and quality of health care indicators. Five main concepts include patient/family context, structure of care, process of care, satisfaction with care, and quality and length of life. Patient and family perspectives are considered. The framework emphasizes quality of life, thus detailed definitions of each quality of life domain are provided. The goal is to facilitate development of a comprehensive set of measures to describe the quality of life of dying persons and evaluate the care they receive, to be used to improve end-of-life care.


Negative regulation of CD45 protein tyrosine phosphatase activity by ionomycin in T cells.
CD45 is a leukocyte-specific, transmembrane protein tyrosine phosphatase (PTPase) required for T cell responsiveness. How the activity of PTPases is regulated in vivo is unclear. Treatment of murine thymocytes and a variety of murine T cell lines with the calcium ionophore ionomycin decreased CD45 PTPase activity. Ionomycin treatment also led to a decreased phosphorylation of serine residues in CD45. These results indicate that increased intracellular calcium modulates CD45 PTPase activity, demonstrating regulation of CD45 PTPase activity in vivo, and also implicate serine dephosphorylation as a possible mechanism.


Questionable methods of cancer management: 'nutritional' therapies.
Although dietary measures may be helpful in preventing certain cancers, there is no scientific evidence that any nutritionally related regimen is appropriate as a primary treatment for cancer. This paper focuses on seven questionable modalities: vitamin C, pau d'arco tea, the Gerson diet, Hoxsey herbal therapy, the macrobiotic diet, Manner metabolic therapy, and Kelley metabolic therapy. Some of these approaches involve a diet that is nutritionally inadequate. Some involve potentially toxic doses of vitamins and/or other substances. Some are quite expensive. All pose the risk that patients who use them will abandon effective treatment. The American Cancer Society therefore recommends that "nutritional cancer cures" be avoided.


Animal milk sustains micronutrient nutrition and child anthropometry among pastoralists in Samburu, Kenya.
Milk has been integral to pastoralist nutrition for thousands of years, but as communities move toward settled livelihoods, milk consumption is dropping with only minimal evidence for the health and nutrition implications. This longitudinal study aimed to first test whether increased dependency on agriculture reduced household milk production and consumption, and ultimately, nutrient adequacy among the Samburu pastoralists. Second, we investigated whether household milk availability affected child milk intakes and anthropometry. Socioeconomic and dietary intake data were collected from households (n = 200) in 2000, 2005, and 2010, and anthropometric measures and individual child milk intakes in 2012. Nutrient intakes were assessed by the probability of nutrient adequacy method, and generalized least-squared regression modeling with mixed effects was applied to identify predictors of milk consumption. Milk contributed 10% of energy intakes, below maize (52%) and sugar (11%), but over one-half of critical micronutrients, vitamins A, B12 , and C. Livestock holdings and income increased the likelihood of higher milk intakes (overall adj R(2) = 0.88, P < 0.001). Undernutrition was widely prevalent among young children: stunting (30.6%); underweight (23.9%); and wasting (8.6%). There was evidence for a previously described Nilotic body type in the youth, who were taller and thinner than the international reference. Milk consumption at the household level was positively associated with higher body mass index z scores among youth (P < 0.001). Programming for livestock development may better ensure micronutrient nutrition in Samburu, while deeper investigation into the diet and growth patterns of pastoralists could provide insight into leaner and taller anthropometrics for other populations globally.


The elongation factor Spt4/5 regulates RNA polymerase II transcription through the nucleosome.
RNA polymerase II (RNAPII) passes through the nucleosome in a coordinated manner, generating several intermediate nucleosomal states as it breaks and then reforms histone-DNA contacts ahead of and behind it, respectively. Several studies have defined transcription-induced nucleosome intermediates using only RNA Polymerase. However, RNAPII is decorated with elongation factors as it transcribes the genome. One such factor, Spt4/5, becomes an integral component of the elongation complex, making direct contact with the 'jaws' of RNAPII and nucleic acids in the transcription scaffold. We have characterized the effect of incorporating Spt4/5 into the elongation complex on transcription through the 601R nucleosome. Spt4/5 suppressed RNAPII pausing at the major H3/H4-induced arrest point, resulting in downstream re-positioning of RNAPII further into the nucleosome. Using a novel single molecule FRET system, we found that Spt4/5 affected the kinetics of DNA re-wrapping and stabilized a nucleosomal intermediate with partially unwrapped DNA behind RNAPII. Comparison of nucleosomes of different sequence polarities suggest that the strength of the DNA-histone interactions behind RNAPII specifies the Spt4/5 requirement. We propose that Spt4/5 may be important to coordinate the mechanical movement of RNAPII through the nucleosome with co-transcriptional chromatin modifications during transcription, which is affected by the strength of histone-DNA interactions.


Hemodynamic, ventilatory and gasometric evaluation of an experimental bronchopleural fistula.
To investigate the hemodynamic and ventilatory changes associated with the creation of an experimental bronchopleural fistula (BPF) treated by mechanical ventilation and thoracic drainage with or without a water seal. Six large white pigs weighing 25 kg each which, after general anesthesia, underwent endotracheal intubation (6mm), and mechanically ventilation. Through a left thoracotomy, a resection of the lingula was performed in order to create a BPF with an output exceeding 50% of the inspired volume. The chest cavity was closed and drained into the water sealed system for initial observation of the high output BPF. Significant reduction in BPF output and PaCO2 was related after insertion of a water-sealed thoracic drain, p< 0.05. Insertion of a water-sealed thoracic drain resulted in reduction in bronchopleural fistula output and better CO2 clearance without any drop in cardiac output or significant changes in mean arterial pressure.


An alpha-proteobacterial type malate dehydrogenase may complement LDH function in Plasmodium falciparum. Cloning and biochemical characterization of the enzyme.
Malate dehydrogenase (MDH) may be important in carbohydrate and energy metabolism in malarial parasites. The cDNA corresponding to the MDH gene, identified on chromosome 6 of the Plasmodium falciparum genome, was amplified by RT-PCR, cloned and overexpressed in Escherichia coli. The recombinant Pf MDH was purified to homogeneity and biochemically characterized as an NAD(+)(H)-specific MDH, which catalysed reversible interconversion of malate to oxaloacetate. Pf MDH could not use NADP/NADPH as a cofactor, but used acetylpyridine adenine dinucleoide, an analogue of NAD. The enzyme exhibited strict substrate and cofactor specificity. The highest levels of Pf MDH transcripts were detected in trophozoites while the Pf MDH protein level remained high in trophozoites as well as schizonts. A highly refined model of Pf MDH revealed distinct structural characteristics of substrate and cofactor binding sites and important amino acid residues lining these pockets. The active site amino acid residues involved in substrate binding were conserved in Pf MDH but the N-terminal glycine motif, which is involved in nucleotide binding, was similar to the GXGXXG signature sequence found in Pf LDH and also in alpha-proteobacterial MDHs. Oxamic acid did not inhibit Pf MDH, while gossypol, which interacts at the nucleotide binding site of oxidoreductases and shows antimalarial activity, inhibited Pf MDH also. Treatment of a synchronized culture of P. falciparum trophozoites with gossypol caused induction in expression of Pf MDH, while expression of Pf LDH was reduced and expression of malate:quinone oxidoreductase remained unchanged. Pf MDH may complement Pf LDH function of NAD/NADH coupling in malaria parasites. Thus, dual inhibitors of Pf MDH and Pf LDH may be required to target this pathway and to develop potential new antimalarial drugs.


The meaning of meaningful use. Several technology applications are needed to qualify.
The American Recovery and Reinvestment Act of 2009 included billions of dollars in Medicare and Medicaid incentive payments to providers and hospitals for the Meaningful Use of certified health IT products. The Act also required the secretary of Health and Human Services (HHS) to establish by the end of 2009 certification criteria and standards for achieving meaningful use. At press time, the final rule for Meaningful Use had not been issued. New information may be available from the Office of the National Coordinator in HHS. This article provides an overview of the Meaningful Use of an EHR and the Interim Final Rule (IFR) establishing the Initial Set of Standards, Implementation Specifications and Certification Criteria for EHR Technology issued in January 2010.


Comparative Evaluation of the Effects of Light Intensities and Curing Cycles of QTH, and LED Lights on Microleakage of Class V Composite Restorations.
The purpose of this study was to evaluate the effects of light intensity and curing cycles of QTH and LED lights on the microleakage of Class V composite restorations. Eighty freshly extracted human maxillary premolars were used for this study. Standardized Class V cavities were prepared and they were restored with microhybrid resin composite. According to the curing protocol, the teeth were then divided into 4 groups (n=20): QTH curing (standard and soft start modes), and LED (standard and soft start modes) irradiations. Microleakage was evaluated by immersion of the samples in 50% silver nitrate solution. The samples were then sectioned, evaluated under a stereomicroscope, and scored for microleakage. The results of the present in-vitro study showed mean dye leakage scores of 1.9, 1.2, 1.45 and 0.90 for Group I (QTH-Standard mode), Group II (QTH-Soft Start mode), Group III (LED-Standard mode) and Group IV (LED-Soft Start mode) respectively. It was thus concluded that the soft start polymerization showed a highly significant difference as compared to the standard curing modes of QTH and LED lights, respectively.


Fasting plasma glucose and risk of incident ischemic stroke or transient ischemic attacks: a prospective cohort study.
Diabetes and impaired fasting glucose are diagnosed based on an elevated plasma glucose level after an overnight fast. The diagnostic cutpoint of diabetes arises from the threshold for development of microvascular complications. Our aim was to examine the associations between clinical relevant categories of fasting glucose levels and the risk of incident ischemic stroke. Patients with documented coronary heart disease who were screened for inclusion in a secondary prevention clinical trial (n=13,999) were followed-up. At baseline, medical histories were obtained and plasma glucose and lipids assessed at a central study laboratory. During a 6- to 8-year follow-up period 1037 cases were identified with ischemic cerebrovascular disease, of which, after reviewing hospital records with diagnoses of cerebrovascular disease, 576 cases were verified to have had ischemic stroke or transient ischemic attacks. Increasing fasting glucose level categories were positively associated with increasing age, male gender, body mass index, hypertension, total cholesterol, and triglycerides, and were inversely associated with high-density lipoprotein cholesterol and percent high-density lipoprotein of total cholesterol. In comparison with patients with fasting glucose levels of 90 to 99 mg/dL (n=3706) who constitute the largest category, the odds ratios of ischemic cerebrovascular disease, adjusting for potential confounders, were 1.47 (95% CI, 1.07 to 2.02) for fasting glucose <80, 1.22 (0.98 to 1.52) for 80 to 89, 1.27 (1.02 to 1.60) for 100 to 109, 1.60 (1.26 to 2.03) for 110 to 125, 1.82 (1.33 to 2.49) for 126 to 140, and 2.82 (2.32 to 3.43) for >140 mg/dL. Similar J-shaped associations were observed in analysis excluding patients with known diagnosis of diabetes mellitus. The association between fasting plasma glucose and incident ischemic cerebrovascular events in patients with pre-existing atherothrombotic disease is J-shaped. Rates increase for fasting plasma glucose levels >100 mg/dL and also for those with low fasting glucose levels. These findings may carry important implications for prevention strategies.


Use of recombinant retroviruses to characterize the activity of antiretroviral compounds.
This report describes the use of a recombinant murine retrovirus encoding beta-galactosidase (PLJ beta-gal retrovirus) to study the antiretroviral activity of zidovudine (AZT) and other nucleoside analogs. The PLJ beta-gal virus permits the rapid and unequivocal identification of individual virus-infected cells arising from a single cycle of viral replication. With this model system, AZT is shown to completely and irreversibly prevent retrovirus infection of proliferating cell lines as measured by a lack of reporter gene expression. On the other hand, AZT is less effective in protecting growth-arrested cells from retroviral infection. Recombinant retroviruses such as the PLJ beta-gal virus are potentially useful reagents for the identification and characterization of antiretroviral compounds.


Trends in the hospital financial picture.
In 1992 (the most recent year for which complete financial data are available), the overall financial condition of U.S. hospitals was better than it had been in a decade. However, low-performing hospitals were falling further behind their high-performing counterparts. Moreover, the continuing drop in prices for services, which requires hospitals to achieve greater productivity and cost control, may pose a threat to the ability of some to maintain high-quality care in the future.


PR130 is a modulator of the Wnt-signaling cascade that counters repression of the antagonist Naked cuticle.
The Wnt-signaling cascade is required for several crucial steps during early embryogenesis, and its activity is modulated by various agonists and antagonists to provide spatiotemporal-specific signaling. Naked cuticle is a Wnt antagonist that itself is induced by Wnt signaling to keep Wnt signaling in check. Little is known about the regulation of this antagonist. We have recently shown that the protein phosphatase 2A regulatory subunit PR72 is required for the inhibitory effect of Naked cuticle on Wnt signaling. In the present study, we show that PR130, which has an N terminus that differs from that of PR72 but shares the same C terminus, also interacts with Naked cuticle but instead functions as an activator of the Wnt-signaling pathway, both in cell culture and during development. We find that PR130 modulates Wnt signal transduction by restricting the ability of Naked cuticle to function as a Wnt inhibitor. Our data establish PR130 as a modulator of the Wnt-signaling pathway and suggest a mechanism of Wnt signal regulation in which the inhibitory activity of Naked cuticle is determined by the relative level of expression of two transcripts of the same protein phosphatase 2A regulatory subunit.


SUPT6H controls estrogen receptor activity and cellular differentiation by multiple epigenomic mechanisms.
The estrogen receptor alpha (ERα) is the central transcriptional regulator of ductal mammary epithelial lineage specification and is an important prognostic marker in human breast cancer. Although antiestrogen therapies are initially highly effective at treating ERα-positive tumors, a large number of tumors progress to a refractory, more poorly differentiated phenotype accompanied by reduced survival. A better understanding of the molecular mechanisms involved in the progression from estrogen-dependent to hormone-resistant breast cancer may uncover new targets for treatment and the discovery of new predictive markers. Recent studies have uncovered an important role for transcriptional elongation and chromatin modifications in controlling ERα activity and estrogen responsiveness. The human Suppressor of Ty Homologue-6 (SUPT6H) is a histone chaperone that links transcriptional elongation to changes in chromatin structure. We show that SUPT6H is required for estrogen-regulated transcription and the maintenance of chromatin structure in breast cancer cells, possibly in part through interaction with RNF40 and regulation of histone H2B monoubiquitination (H2Bub1). Moreover, we demonstrate that SUPT6H protein levels decrease with malignancy in breast cancer. Consistently, SUPT6H, similar to H2Bub1, is required for cellular differentiation and suppression of the repressive histone mark H3K27me3 on lineage-specific genes. Together, these data identify SUPT6H as a new epigenetic regulator of ERα activity and cellular differentiation.


Motor coordination and psychosocial correlates in a normative adolescent sample.
Previous research has revealed an important relationship between motor coordination difficulties and internalizing problems such as anxiety and depressive symptoms. However, further research is needed to understand the potential mediating factors in this relationship. The aim of the current study was to examine whether the association between motor coordination and emotional functioning is mediated by self-perceptions in a normative sample of adolescents. Participants included 93 adolescents aged 12 to 16 years. The Movement Assessment Battery for Children-2 provided 2 indicators of motor coordination; the Mood and Feelings Questionnaire and Spence Children's Anxiety Scale provided 2 indicators of emotional functioning; and the Self-Description Questionnaire--II provided 6 indicators for self-perceived competence. Structural equation modeling revealed that motor coordination affects emotional functioning via self-perceptions. These results suggest that the relationship between motor coordination and emotional functioning in adolescents from a normative sample may be understood in terms of a mechanism by which motor coordination has an indirect impact on emotional outcomes through various self-perception domains. These findings have important implications for increasing awareness and developing appropriate treatment programs for motor coordination and emotional difficulties.


Staphylococcus aureus bacteremia (SAB) with associated S. aureus bacteriuria (SABU) as a predictor of complications and mortality.
Staphylococcus aureus (SA) bacteremia (SAB) is associated with a high rate of complications, most of which are related to hematogenous seeding into deep tissues or prosthetic material. SA bacteriuria (SABU) has been described in association with SAB, but has not been evaluated as a predictor for complicated bacteremia, which was the objective of our study. METHODS (DESIGN, SETTING, AND PATIENTS): We conducted a retrospective study of patients admitted to the hospital with SAB. The 118 patients included in the study were divided in 2 cohorts: a group with SABU and a group without SA in the urine. We followed the 2 cohorts for an average of 8 months and evaluated the differences in complications and mortality. SABU was found in 28 of 118 patients with SAB. Eighteen patients (64%) in this group had complications from the bacteremia, while in the group without SABU only 33% (30/90 patients) had complications (P = 0.004). The SABU group also had more deaths (32% vs. 14%; P = 0.036). In this population of hospitalized patients with SAB, the presence of SABU was associated with an increased risk of early complications, including septic shock, and with higher mortality. A routine urine culture in search of SABU may be a helpful tool for detection of those patients with SAB who are at increased risk of complications and death.


Wear Mechanism Classification Using Artificial Intelligence.
Understanding the acting wear mechanisms in many cases is key to predicting lifetime, developing models describing component behavior, or for the improvement of the performance of components under tribological loading. Conventionally scanning electron microscopy (SEM) and sometimes additional analytical techniques are performed in order to analyze wear appearances, i.e., grooves, pittings, surface films, and others. In addition, experience is required in order to draw the correct and relevant conclusions on the acting damage and wear mechanisms from the obtained analytical data. Until now, different types of wear mechanisms are classified by experts examining the damage patterns manually. In addition to this approach based on expert knowledge, the use of artificial intelligence (AI) represents a promising alternative. Here, no expert knowledge is required, instead, the classification is done by a purely data-driven model. In this contribution, artificial neural networks are used to classify the wear mechanisms based on SEM images. In order to obtain optimal performance of the artificial neural network, a hyperparameter optimization is performed in addition. The content of this contribution is the investigation of the feasibility of an AI-based model for the automated classification of wear mechanisms.


Interactions between beta-2 adrenoceptor gene variation, cardiovascular control and dietary sodium in healthy young adults.
Dietary sodium affects function of the beta-2 adrenoceptor (ADRB2). We tested the hypothesis that haplotype variation in the ADRB2 gene would influence the cardiovascular and regional vasodilator responses to sympathoexcitatory manoeuvres following low, normal and high sodium diets, and ADRB2-mediated forearm vasodilation in the high sodium condition. Seventy-one healthy young adults were grouped by double homozygous haplotypes: Arg16+Gln27 (n = 31), the rare Gly16+Gln27 (n = 10) and Gly16+Glu27 (n = 30). Using a randomized cross-over design, subjects were studied following 5 days of controlled low, normal and high sodium with 1 month or longer between diets (and low hormone phase of the menstrual cycle). All three visits utilized ECG and finger plethysmography for haemodynamic measures, and the high sodium visit included a brachial arterial catheter for forearm vasodilator responses to isoprenaline with plethysmography. Lymphocytes were sampled for ex vivo analysis of ADRB2 density and binding conformation. We found a main effect of haplotype on ADRB2 density (P = 0.03) with the Gly16+Glu27 haplotype having the greatest density (low, normal, high sodium: 12.9 ± 0.9, 13.5 ± 0.9 and 13.6 ± 0.8 fmol mg(-1) protein, respectively) and Arg16+Gln27 having the least (9.3 ± 0.6, 10.1 ± 0.5 and 10.3 ± 0.6 fmol mg(-1) protein, respectively), but there were no sodium or haplotype effects on receptor binding conformation. In the mental stress trial, there was a main effect of haplotype on cardiac output (P = 0.04), as Arg16+Gln27 had the lowest responses. Handgrip and forearm vasodilation yielded no haplotype differences, and no correlations were present for ADRB2 density and haemodynamics. Our findings support cell-based evidence that ADRB2 haplotype influences ADRB2 protein expression independent of dietary sodium, yet the haemodynamic consequences appear modest in healthy humans.


Phosphate release from waste stabilisation pond sludge: significance and fate of polyphosphate.
Net phosphorus removal from waste stabilisation pond (WSP) systems is governed by the rate of phosphorus incorporation into the sludge layer and the rate of phosphorus release from this sludge back to the overlying wastewater. Luxury uptake of phosphorus by microalgae has been shown to occur under WSP conditions in the laboratory; however, the significance of this mechanism and the fate of polyphosphate contained in the settled solids have not previously been investigated. In this work the analysis of sludge samples from three WSP showed that up to 71% of the total phosphorus in the sludge was in the form of polyphosphate. This indicates that polyphosphate accumulation could potentially be an important mechanism for phosphorus sequestration in WSP and challenges the common view that chemical precipitation is the predominant phosphorus removal mechanism in these systems. The release of phosphate from WSP sludge samples was monitored in the laboratory. The samples from two different pond systems had release rates in the order of 4.3 microgP/gTSS.d. However, the third sample which was collected during an algal bloom had a release rate of 12.4 microgP/gTSS.d. Phosphate release from fresh microalgal sludge grown under laboratory conditions was also studied and was shown to have a release rate of 160 microgP/gTSS.d. Analysis of polyphosphate during the experiments on laboratory grown microalgal sludge showed that polyphosphate was indeed degraded resulting in phosphate release. Interestingly, after the initial release phase phosphorus was assimilated by the biomass and some polyphosphate was reformed. It is likely that this is due to bacterial growth in the sludge.


Biochemical and molecular characterization of cowpea landraces using seed storage proteins and SRAP marker patterns.
Seven landraces of cowpea [Vigna unguiculata (L.) Walp.] were assessed for genetic variability in total proteins, protein fractions viz. albumins, globulins, prolamins, and glutelins by SDS-polyacrylamide gel electrophoresis and DNA polymorphism using sequence-related amplified polymorphisms (SRAP) markers. The solubility-based protein fractionation data indicated that the salt soluble fraction (globulin) and water-soluble fraction (albumin) proteins were the predominant fractions in cowpea seeds comprising 45-50.3% and 31.2-35.5% of total soluble proteins, respectively. The electrophoretic pattern revealed the molecular heterogeneity among total proteins as well as different protein fractions. The molecular weights of protein bands obtained by SDS-PAGE varied between 10 to 250, 15 to 110, 15 to 150, and 15 to 130 kDa for total proteins, albumins, globulins, and glutelins, respectively. A large number of bands were found common to the various landraces, indicative of their close relationship with one another. However, a few bands distinctive to some specific landraces were also detected, indicating varietal differences. A 34 SRAP primer pair combination generated a total of 1003 amplicons (loci) showed 100% polymorphism with an average of 0.93 polymorphism information content (PIC) value. Landraces displayed an average 0.50 similarity coefficient which clustered the landraces corresponding to their growth habit in main clusters and to their geographical origin in subcultures. Molecular and biochemical analysis were correlated with a medium level (Mantel test, r = 0.56, P < 0.02). These findings revealed that seed proteins and DNA polymorphism provide valuable information regarding the variability among landraces and this information could be utilized for breeding purposes in the enhancement of protein quality and quantity in grain legumes.


Cloning and expression of Candida guilliermondii xylose reductase gene (xyl1) in Pichia pastoris.
A xylose reductase gene (xyl1) of Candida guilliermondii ATCC 20118 was cloned and characterized. The open reading frame of xyl1 contained 954 nucleotides encoding a protein of 317 amino acids with a predicted molecular mass of 36 kDa. The derived amino acid sequence of C. guilliermondii xylose reductase was 70.4% homologous to that of Pichia stipitis. The gene was placed under the control of an alcohol oxidase promoter (AOX1) and integrated into the genome of a methylotrophic yeast, Pichia pastoris. Methanol induced the expression of the 36-kDa xylose reductase in both intracellular and secreted expression systems. The expressed enzyme preferentially utilized NADPH as a cofactor and was functional both in vitro and in vivo. The different cofactor specificity between P. pastoris and C. guilliermondii xylose reductases might be due to the difference in the numbers of histidine residues and their locations between the two proteins. The recombinant was able to ferment xylose, and the maximum xylitol accumulation (7.8 g/l) was observed when the organism was grown under aerobic conditions.


The Efficacy of the Geriatric Model of Care in Emergency Housing Programs for Homeless Veterans.
The aging homeless population currently makes up half the general homeless population. However, there are few homeless emergency shelters that can address their needs. This results in an overutilization of inpatient admissions and emergency room services. Homeless service staff from VA Palo Alto Health Care System partnered with a local homeless emergency housing provider, Compassion Residio Services Inc., to implement this new model of care for aging, medically fragile homeless Veterans. This emergency housing model utilizes practices done in geriatric settings. This model aimed to help decrease the utilization of emergency departments and inpatient admissions. The average cost of emergency department visits and inpatient admissions was $127,314 per Veteran 6 months before admission. Six months after discharge, the average cost of treatment from emergency department visits and inpatient admissions was roughly $59,546 after discharge, a 53% decrease. Emergency department visits decreased from an average of 5.6 visits per Veteran 6 months before admission to 2.65 visits after 6 months discharge. The number of inpatient nights decreased from an average of 15 days per admission in the 6 months before the program to 13 days. Total admissions decreased by nearly half from 48 the previous 6 months to 25 after 6 months. Overall, as residents settled into stable environments tailored around geriatric care, the utilization of emergency department visits and inpatient services decreased. Furthermore, the complexity (eg, cost per encounter) also decreased.


Identified Aplysia neurons form specific chemical synapses in culture.
Identified neurons from the abdominal ganglion of the marine mollusc Aplysia californica make specific transmitter-mediated synapses in dissociated cell culture. The cholinergic interneuron L10 makes synapses in vitro with one group of its follower cells, the left upper quadrant cells, and these connections exhibit the features of these synapses in vivo when the postsynaptic cells are plated with their initial axon segments. Furthermore, L10 will avoid making synapses with right upper quadrant cells, which contain cholinergic receptors but do not synapse with L10 in vivo. This in vitro system can therefore be used as a model in which to study the development of specific neuronal connections.


A Rare Case of Pancolonic Ischemia Complicated by Fusobacterium Necrophorum Bacteremia: A Review of Colonic Ischemia for Internists.
The varied clinical presentation and objective findings associated with colonic ischemia (CI) overlap with multiple disease processes. A high index of suspicion is critical for timely diagnosis and prognostication to avoid delays in treatment. We present a case that highlights the challenges of diagnosing CI and the high morbidity associated with severe disease. Case report: A sixty-four-year-old female presented to our community hospital with acute onset abdominal pain, nausea, and diarrhea, complicated by septic shock. She was initially given a diagnosis of infectious colitis until a colonoscopy was performed revealing extensive pancolonic ulcerations. Histopathological features on biopsy were most consistent with colonic ischemia. Blood cultures grew Fusobacterium Necrophorum. Surgery was avoided due to the high morbidity and mortality of performing a total colectomy and ileostomy. The patient continued to struggle with abdominal pain, diarrhea, and hematochezia, consistent with continuous CI, leading to recurrent hospitalizations.


Depression in People With Dementia and Caregiver Outcomes: Results From the European Right Time Place Care Study.
To investigate the cross-sectional associations between depression in people with dementia and both caregiver burden and quality of life in 8 European countries, and to test these associations compared with the presence of other neuropsychiatric symptoms. Cross-sectional study. In total, 1223 dyads comprised of informal caregivers and people with dementia living in a community-dwelling setting, recruited from the Right Time Place Care study, a cohort survey from 8 European countries. To test the associations between depression (according to the Cornell Scale for Depression in Dementia) and informal caregiver burden (defined by the Zarit scale and hours of supervision in terms of Resource Utilization in Dementia), distress (defined by the Neuropsychiatric Inventory Questionnaire distress score), and quality of life (according to the visual analogue scale and 12-item General Health Questionnaire). Linear regressions showed an association between depression and main outcomes (Zarit scale: β 3.7; P = .001; hours of supervision: β 1.7; P = .004; Neuropsychiatric Inventory Questionnaire distress score: β 1.2; P = .002). A similar association was found concerning psychological and overall well-being (12-item General Health Questionnaire: β 1.8; P < .001; Euroqol Visual Analogue Scale: β -4.1; P = .003). Both associations remained significant despite the presence of other NPS and after adjusting for confounders. Further studies are needed to assess whether providing tailored strategies for optimizing diagnosis and managing of depression in people with dementia might improve caregiver quality of life and reduce their burden in the community-dwelling setting.


Repurposing psychiatric medicines to target activated microglia in anxious mild cognitive impairment and early Parkinson's disease.
Anxiety is common in the Mild Cognitive Impairment (MCI) stage of Alzheimer's disease (AD) and the pre-motor stages of Parkinson's disease (PD). A concomitant and possible cause of this anxiety is microglial activation, also considered a key promoter of neurodegeneration in MCI and early PD via inflammatory mechanisms and the generation of degenerative proinflammatory cytokines. Psychiatric disorders, prevalent in AD and PD, are often treated with psychiatric drugs (psychotropics), raising the question of whether psychotropics might therapeutically affect microglial activation, MCI, and PD. The literature of common psychotropics used in treating psychiatric disorders was reviewed for preclinical and clinical findings regarding microglial activation. Findings potentially compatible with reduced microglial activation or reduced microglial inflammogen release were evident for: antipsychotics including neuroleptics (chlorpromazine, thioridazine, loxapine) and atypicals (aripiprazole, olanzapine, quetiapine, risperidone, ziprasidone); mood stabilizers (carbamazepine, valproate, lithium); antidepressants including tricyclics (amitriptyline, clomipramine, imipramine, nortriptyline), SSRIs (citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine, sertraline), venlafaxine, and bupropion; benzodiazepine anxiolytics (clonazepam, diazepam); cognitive enhancers (donepezil, galantamine, memantine); and other drugs (dextromethorphan, quinidine, amantadine). In contrast, pramipexole and methylphenidate might promote microglial activation. The most promising replicated findings of reduced microglial activation are for quetiapine, valproate, lithium, fluoxetine, donepezil, and memantine but further study is needed and translation of their microglial effects to human disease still requires investigation. In AD-relevant models, risperidone, valproate, lithium, fluoxetine, bupropion, donepezil, and memantine have therapeutic microglial effects in need of replication. Limited clinical data suggest some support for lithium and donepezil in reducing MCI progression, but other drugs have not been studied. In PD-relevant models, lamotrigine, valproate, fluoxetine, dextromethorphan, and amantadine have therapeutic microglial effects whereas methylphenidate induced microglial activation and pramipexole promoted NO release. Clinical data limited to pramipexole do not as of yet indicate faster progression of early PD while the other drugs remain to be investigated. These tantalizing psychotropic neuroprotective findings now invite replication and evidence in AD-and PD-specific models under chronic administration, followed by consideration for clinical trials in MCI and early stage PD. Psychiatric features in early disease may provide opportunities for clinical studies that also employ microglial PET biomarkers.


AP1- and NF-kappaB-binding sites conserved among mammalian WNT10B orthologs elucidate the TNFalpha-WNT10B signaling loop implicated in carcinogenesis and adipogenesis.
WNT signals are context-dependently transduced to canonical and non-canonical signaling cascades. We cloned and characterized wild-type human WNT10B, while another group cloned aberrant human WNT10B with Gly60Asp amino-acid substitution. Proto-oncogene WNT10B is expressed in gastric cancer, pancreatic cancer, breast cancer, esophageal cancer, and cervical cancer. Because WNT10B blocks adipocyte differentiation, coding SNP of WNT10B gene is associated with familial obesity. In 2001, we reported WNT10B upregulation by TNFalpha. Here, comparative integromics analyses on WNT10B orthologs were performed to elucidate the transcriptional mechanism of WNT10B. Chimpanzee WNT10B and cow Wnt10b genes were identified within NW_001223159.1 and AC150975.2 genome sequences, respectively, by using bioinformatics (Techint) and human intelligence (Humint). Chimpanzee WNT10B and cow Wnt10b showed 98.7% and 95.1% total-amino-acid identity with human WNT10B, respectively. N-terminal signal peptide, 24 Cys residues, two Asn-linked glycosylation sites, and Gly60 of human WNT10B were conserved among mammalian WNT10B orthologs. Transcription start site of human WNT10B gene was 106-bp upstream of NM_003394.2 RefSeq 5'-end. Number of GC di-nucleotide repeats just down-stream of WNT10B transcription start site varied among primates and human population. Comparative genomics analyses revealed that double AP1-binding sites in the 5'-flanking promoter region and NF-kappaB-binding site in intron 3 were conserved among human, chimpanzee, cow, mouse, and rat WNT10B orthologs. Because TNFalpha signaling through TNFR1 and TRADD/RIP/TRAF2 complex activates JUN kinase (JNK) and IkappaB kinase (IKK) signaling cascades, conserved AP1- and NF-kappaB-binding sites explain the mechanism of TNFalpha-induced WNT10B upregulation. TNFalpha-WNT10B signaling loop is the negative feedback mechanism of adipogenesis to prevent obesity and metabolic syndrome. On the other hand, TNFalpha-WNT10B signaling loop is implicated in carcinogenesis. Inhibitors of TNFalpha-WNT10B signaling loop could be utilized for the prevention or treatment of cancer associated with chronic inflammation, such as gastric, liver, breast and pancreatic cancer.


Vitamin D Status Is Associated With In-Hospital Mortality and Mechanical Ventilation: A Cohort of COVID-19 Hospitalized Patients.
To explore the possible associations of serum 25-hydroxyvitamin D [25(OH)D] concentration with coronavirus disease 2019 (COVID-19) in-hospital mortality and need for invasive mechanical ventilation. A retrospective, observational, cohort study was conducted at 2 tertiary academic medical centers in Boston and New York. Eligible participants were hospitalized adult patients with laboratory-confirmed COVID-19 between February 1, 2020, and May 15, 2020. Demographic and clinical characteristics, comorbidities, medications, and disease-related outcomes were extracted from electronic medical records. The final analysis included 144 patients with confirmed COVID-19 (median age, 66 years; 64 [44.4%] male). Overall mortality was 18%, whereas patients with 25(OH)D levels of 30 ng/mL (to convert to nmol/L, multiply by 2.496) and higher had lower rates of mortality compared with those with 25(OH)D levels below 30 ng/mL (9.2% vs 25.3%; P=.02). In the adjusted multivariable analyses, 25(OH)D as a continuous variable was independently significantly associated with lower in-hospital mortality (odds ratio, 0.94; 95% CI, 0.90 to 0.98; P=.007) and need for invasive mechanical ventilation (odds ratio, 0.96; 95% CI, 0.93 to 0.99; P=.01). Similar data were obtained when 25(OH)D was studied as a continuous variable after logarithm transformation and as a dichotomous (<30 ng/mL vs ≥30 ng/mL) or ordinal variable (quintiles) in the multivariable analyses. Among patients admitted with laboratory-confirmed COVID-19, 25(OH)D levels were inversely associated with in-hospital mortality and the need for invasive mechanical ventilation. Further observational studies are needed to confirm these findings, and randomized clinical trials must be conducted to assess the role of vitamin D administration in improving the morbidity and mortality of COVID-19.


Molecular toolbox for the identification of unknown genetically modified organisms.
Competent laboratories monitor genetically modified organisms (GMOs) and products derived thereof in the food and feed chain in the framework of labeling and traceability legislation. In addition, screening is performed to detect the unauthorized presence of GMOs including asynchronously authorized GMOs or GMOs that are not officially registered for commercialization (unknown GMOs). Currently, unauthorized or unknown events are detected by screening blind samples for commonly used transgenic elements, such as p35S or t-nos. If (1) positive detection of such screening elements shows the presence of transgenic material and (2) all known GMOs are tested by event-specific methods but are not detected, then the presence of an unknown GMO is inferred. However, such evidence is indirect because it is based on negative observations and inconclusive because the procedure does not identify the causative event per se. In addition, detection of unknown events is hampered in products that also contain known authorized events. Here, we outline alternative approaches for analytical detection and GMO identification and develop new methods to complement the existing routine screening procedure. We developed a fluorescent anchor-polymerase chain reaction (PCR) method for the identification of the sequences flanking the p35S and t-nos screening elements. Thus, anchor-PCR fingerprinting allows the detection of unique discriminative signals per event. In addition, we established a collection of in silico calculated fingerprints of known events to support interpretation of experimentally generated anchor-PCR GM fingerprints of blind samples. Here, we first describe the molecular characterization of a novel GMO, which expresses recombinant human intrinsic factor in Arabidopsis thaliana. Next, we purposefully treated the novel GMO as a blind sample to simulate how the new methods lead to the molecular identification of a novel unknown event without prior knowledge of its transgene sequence. The results demonstrate that the new methods complement routine screening procedures by providing direct conclusive evidence and may also be useful to resolve masking of unknown events by known events.


Serotype 19f multiresistant pneumococcal clone harboring two erythromycin resistance determinants (erm(B) and mef(A)) in South Africa.
One hundred eighteen erythromycin-resistant Streptococcus pneumoniae (ERSP) strains (MICs of > or = 0.5 microg/ml) from five laboratories serving the private sector in South Africa were analyzed for the genes encoding resistance to macrolides. Sixty-seven ERSP strains (56.8%) contained the erm(B) gene, and 15 isolates (12.7%) contained the mef(A) gene. Thirty-six isolates (30.5%) harbored both the erm(B) and mef(A) genes and were highly resistant to erythromycin and clindamycin. DNA fingerprinting by BOX-PCR and pulsed-field gel electrophoresis identified 83% of these strains as belonging to a single multiresistant serotype 19F clone.


Aerobic Interval Training Reduces the Burden of Atrial Fibrillation in the Short Term: A Randomized Trial.
Exercise training is an effective treatment for important atrial fibrillation (AF) comorbidities. However, a high level of endurance exercise is associated with an increased AF prevalence. We assessed the effects of aerobic interval training (AIT) on time in AF, AF symptoms, cardiovascular health, and quality of life in AF patients. Fifty-one patients with nonpermanent AF were randomized to AIT (n=26) consisting of four 4-minute intervals at 85% to 95% of peak heart rate 3 times a week for 12 weeks or to a control group (n=25) continuing their regular exercise habits. An implanted loop recorder measured time in AF continuously from 4 weeks before to 4 weeks after the intervention period. Cardiac function, peak oxygen uptake (o2peak), lipid status, quality of life, and AF symptoms were evaluated before and after the 12-week intervention period. Mean time in AF increased from 10.4% to 14.6% in the control group and was reduced from 8.1% to 4.8% in the exercise group (P=0.001 between groups). AF symptom frequency (P=0.006) and AF symptom severity (P=0.009) were reduced after AIT. AIT improved o2peak, left atrial and ventricular ejection fraction, quality-of-life measures of general health and vitality, and lipid values compared with the control group. There was a trend toward fewer cardioversions and hospital admissions after AIT. AIT for 12 weeks reduces the time in AF in patients with nonpermanent AF. This is followed by a significant improvement in AF symptoms, o2peak, left atrial and ventricular function, lipid levels, and QoL. URL: http://www.clinicaltrials.gov. Unique identifier: NCT01325675.


Adaptation of Campylobacter jejuni NCTC11168 to high-level colonization of the avian gastrointestinal tract.
The genome sequence of the human pathogen Campylobacter jejuni NCTC11168 has been determined recently, but studies on colonization and persistence in chickens have been limited due to reports that this strain is a poor colonizer. Experimental colonization and persistence studies were carried out with C. jejuni NCTC11168 by using 2-week-old Light Sussex chickens possessing an acquired natural gut flora. After inoculation, NCTC11168 initially colonized the intestine poorly. However, after 5 weeks we observed adaptation to high-level colonization, which was maintained after in vitro passage. The adapted strain exhibited greatly increased motility. A second strain, C. jejuni 11168H, which had been selected under in vitro conditions for increased motility (A. V. Karlyshev, D. Linton, N. A. Gregson, and B. W. Wren, Microbiology 148:473-480, 2002), also showed high-level intestinal colonization. The levels of colonization were equivalent to those of six other strains, assessed under the same conditions. There were four mutations in C. jejuni 11168H that reduced colonization; maf5, flaA (motility and flagellation), and kpsM (capsule deficiency) eliminated colonization, whereas pglH (general glycosylation system deficient) reduced but did not eliminate colonization. This study showed that there was colonization of the avian intestinal tract by a Campylobacter strain having a known genome sequence, and it provides a model for colonization and persistence studies with specific mutations.


Canadian Association of Gastroenterology position statement on screening individuals at average risk for developing colorectal cancer: 2010.
The Canadian Association of Gastroenterology and the Canadian Digestive Health Foundation published guidelines on colon cancer screening in 2004. Subsequent to the publication of these guidelines, many advances have occurred, thereby necessitating a review of the existing guidelines in the context of new technologies and clinical knowledge. The assembled guideline panel recognized three recent American sets of guidelines and identified seven issues that required comment from a Canadian perspective. These issues included, among others, the role of program-based screening, flexible sigmoidoscopy, computed tomography colonography, barium enema and quality improvement. The panel also provided context for the selection of the fecal immunochemical test as the fecal occult blood test of choice, and the relative role of colonoscopy as a primary screening tool. Recommendations were also provided for an upper age limit for colon cancer screening, whether upper endoscopy should be performed following a negative colonoscopy for a positive fecal occult blood test and when colon cancer screening should resume following negative colonoscopy.


Is pseudophakic astigmatism a desirable goal?
To determine whether pseudophakic astigmatism is a desirable goal, and if so, which one is better: against-the-rule (ATR) or with-the-rule (WTR). Eyes were included only if they had an uncorrected vision > or = 6/18 and N/18. Three groups, of 40 patients each were evaluated: group 1, pseudophakes with neutral astigmatism; group 2, with ATR and group 3, with WTR astigmatism Unaided distance and near visual acuity was recorded. Statistical analysis was performed using the chi-square test for independence. Unaided distance vision of > or = 6/7.5 was achieved in 19 eyes (47.5%) of group 1 (neutral), 12 eyes (30%) in group 2 (ATR), and 5 eyes (12.5%) in group 3 (WTR) (p = 0.0133, significant). Unaided near vision of > or = N/9 was achieved in 17 eyes (42.5%) in group 1 (neutral), 34 eyes (85%) in group 2 (ATR), and 10 eyes (25%) in group 3 (WTR) (P < 0.001, significant). Group 1 (neutral) fared the best for unaided distance visual acuity. Group 2 (ATR) was better than in group 3 (WTR) for distant vision. Group 2 (ATR) fared the best for unaided near vision. ATR astigmatism could be a desirable goal after cataract extraction in selected populations because the largest proportion of these cases achieved good unaided near vision with acceptable distant vision.


Rapid Cerebral Metabolic Shift during Neonatal Sepsis Is Attenuated by Enteral Colostrum Supplementation in Preterm Pigs.
Sepsis, the clinical manifestation of serious infection, may disturb normal brain development, especially in preterm infants with an immature brain. We hypothesized that neonatal sepsis induces systemic metabolic alterations that rapidly affect metabolic signatures in immature brain and cerebrospinal fluid (CSF). Cesarean-delivered preterm pigs systemically received 10⁸ CFU/kg Staphylococcus epidermidis (SE) and were provided total parenteral nutrition (n = 9) or enteral supplementation with bovine colostrum (n = 10) and compared with uninfected pigs receiving parenteral nutrition (n = 7). Plasma, CSF, and brain tissue samples were collected after 24 h and analyzed by ¹H NMR-based metabolomics. Both plasma and CSF metabolomes revealed SE-induced changes in metabolite levels that reflected a modified energy metabolism. Hence, increased plasma lactate, alanine, and succinate levels, as well as CSF lactate levels, were observed during SE infection (all p < 0.05, ANOVA analysis). Myo-inositol, a glucose derivative known for beneficial effects on lung maturation in preterm infants, was also increased in plasma and CSF following SE infection. Enteral colostrum supplementation attenuated the lactate accumulation in blood and CSF. Bloodstream infection in preterm newborns was found to induce a rapid metabolic shift in both plasma and CSF, which was modulated by colostrum feeding.


Family resilience and adaptive coping in children with juvenile idiopathic arthritis: protocol for a systematic review.
This systematic review is the first step in a study investigating the resilience methods and processes in families of children with juvenile idiopathic arthritis. In particular, this review will focus on chronic or persistent pain, as a common symptom of juvenile idiopathic arthritis, which is the most common rheumatic disease in childhood. The experience of persistent pain can add to the functional disability associated with juvenile idiopathic arthritis. Resilience has relevance to all areas of paediatric psychology, and targeted attention to child, sibling, and parent strengths within the context of paediatric chronic pain and juvenile idiopathic arthritis in particular will augment the field on numerous levels. The objective is to determine which resilience processes are associated with a favourable quality of life in terms of academic, communication, emotional, interpersonal, physical, psychological, and social well-being in families of children with chronic pain associated with JIA. This systematic review will be conducted and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement and the PRESS (Peer Review of Electronic Search Strategies) guideline. Longitudinal, cross-sectional, and treatment studies written in English will be included, as will grey literature (i.e. conference abstracts and dissertations). Studies involving participants who are 6-18 years of age, have been diagnosed with juvenile idiopathic arthritis, are experiencing chronic pain, and are currently undergoing treatment will be included regardless of sex, arthritis type, and type of treatment. Studies including siblings who are 6-18 years of age and the patient's parents will be included. Research exploring resilience within the adult population is accruing. Shifting our focus to protective factors of resilience in the context of paediatric chronic pain, specifically juvenile idiopathic arthritis, is a novel and promising pursuit that has the potential to optimize and inform future clinical practice and interventions. A better understanding of the role of reliance in family adaptation will facilitate the development of more effective treatment approaches and lay the foundation for more effective self-management in paediatric chronic pain. This protocol is registered with the International Prospective Register of Systematic Reviews (PROSPERO) database (registration: CRD42016047226).


DNA Origami Nanoarrays for Multivalent Investigations of Cancer Cell Spreading with Nanoscale Spatial Resolution and Single-Molecule Control.
We present a strategy for the fabrication of biomimetic nanoarrays, based on the use of DNA origami, that permits the multivalent investigation of ligand-receptor molecule interactions in cancer cell spreading, with nanoscale spatial resolution and single-molecule control. We employed DNA origami to control the nanoscale spatial organization of integrin- and epidermal growth factor (EGF)-binding ligands that modulate epidermal cancer cell behavior. By organizing these multivalent DNA nanostructures in nanoarray configurations on nanopatterned surfaces, we demonstrated the cooperative behavior of integrin and EGF ligands in the spreading of human cutaneous melanoma cells: this cooperation was shown to depend on both the number and ratio of the selective ligands employed. Notably, the multivalent biochips we have developed allowed for this cooperative effect to be demonstrated with single-molecule control and nanoscale spatial resolution. By and large, the platform presented here is of general applicability for the study, with molecular control, of different multivalent interactions governing biological processes from the function of cell-surface receptors to protein-ligand binding and pathogen inhibition.


Denitrifying community in coastal sediments performs aerobic and anaerobic respiration simultaneously.
Nitrogen (N) input to the coastal oceans has increased considerably because of anthropogenic activities, however, concurrent increases have not occurred in open oceans. It has been suggested that benthic denitrification in sandy coastal sediments is a sink for this N. Sandy sediments are dynamic permeable environments, where electron acceptor and donor concentrations fluctuate over short temporal and spatial scales. The response of denitrifiers to these fluctuations are largely unknown, although previous observations suggest they may denitrify under aerobic conditions. We examined the response of benthic denitrification to fluctuating oxygen concentrations, finding that denitrification not only occurred at high O2 concentrations but was stimulated by frequent switches between oxic and anoxic conditions. Throughout a tidal cycle, in situtranscription of genes for aerobic respiration and denitrification were positively correlated within diverse bacterial classes, regardless of O2 concentrations, indicating that denitrification gene transcription is not strongly regulated by O2 in sandy sediments. This allows microbes to respond rapidly to changing environmental conditions, but also means that denitrification is utilized as an auxiliary respiration under aerobic conditions when imbalances occur in electron donor and acceptor supply. Aerobic denitrification therefore contributes significantly to N-loss in permeable sediments making the process an important sink for anthropogenic N-inputs.


Immunomodulatory effects of Sanglifehrin A in the innate and acquired immune response of neonatal whole blood cells.
Previous investigations have shown the immunosuppressive activity of the immunophilin-binding macrolide Sanglifehrin A (SFA). In adults, SFA also exerts anti-inflammatory properties in lipopolysaccharide (LPS)-stimulated whole blood cultures. It was the aim of this study to investigate whether the unique properties of SFA are also present in the neonatal immune system, as neonates are susceptible to serious infection due to an immaturity of immune responses. We used a whole blood assay to investigate the impact of SFA on T-cell proliferation and secretion of T-cell cytokines upon Anti-CD3/Anti-CD28 costimulation. In addition, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) secretion was assessed in whole blood monocytes after LPS stimulation. Furthermore, the influence of SFA on LPS-induced signal transduction pathways, specifically the activity of p42/44, p38 and Ap-1, was assessed in neonatal PBMCs. Neonatal cord blood lymphocytes were found to have a diminished IL-4, IL-6, TNF-alpha and interferon-gamma (IFN-gamma) production upon Anti-CD3/Anti-CD28 costimulation compared to adult whole blood lymphocytes. In contrast, no significant differences were noted for either IL-2 production or proliferation of CD4+ cells. Upon addition of 1000nM SFA to neonatal whole blood cultures, a significant inhibition of both, T-cell cytokine secretion and proliferation was demonstrated. In line with data from adult whole blood cultures, SFA proved to be a strong inhibitor of LPS-induced expression of IL-6 and TNF-alpha in the neonatal whole blood system. In signal transduction studies of the LPS pathway, a potent inhibition of the protein kinase p42/44 was demonstrated. SFA was also shown to block nuclear translocation of the transcription factor Ap-1. SFA was proved to have inhibitory effects on innate and acquired immune response of neonatal whole blood cells. The protein kinase p42/44 and the transcription factor Ap-1 were demonstrated to be potential key molecules for the anti-inflammatory effect of SFA.


Evidence-based risk assessment and communication: a new global dengue-risk map for travellers and clinicians.
International travel can expose travellers to pathogens not commonly found in their countries of residence, like dengue virus. Travellers and the clinicians who advise and treat them have unique needs for understanding the geographic extent of risk for dengue. Specifically, they should assess the need for prevention measures before travel and ensure appropriate treatment of illness post-travel. Previous dengue-risk maps published in the Centers for Disease Control and Prevention's Yellow Book lacked specificity, as there was a binary (risk, no risk) classification. We developed a process to compile evidence, evaluate it and apply more informative risk classifications. We collected more than 839 observations from official reports, ProMED reports and published scientific research for the period 2005-2014. We classified each location as frequent/continuous risk if there was evidence of more than 10 dengue cases in at least three of the previous 10 years. For locations that did not fit this criterion, we classified locations as sporadic/uncertain risk if the location had evidence of at least one locally acquired dengue case during the last 10 years. We used expert opinion in limited instances to augment available data in areas where data were sparse. Initial categorizations classified 134 areas as frequent/continuous and 140 areas as sporadic/uncertain. CDC subject matter experts reviewed all initial frequent/continuous and sporadic/uncertain categorizations and the previously uncategorized areas. From this review, most categorizations stayed the same; however, 11 categorizations changed from the initial determinations. These new risk classifications enable detailed consideration of dengue risk, with clearer meaning and a direct link to the evidence that supports the specific classification. Since many infectious diseases have dynamic risk, strong geographical heterogeneities and varying data quality and availability, using this approach for other diseases can improve the accuracy, clarity and transparency of risk communication.


Moral case deliberation.
Ethical dilemmas in general are characterised by a choice between two mutually excluding options neither of which is satisfactory, because there always will be a form of moral damage. Within the context of medicine several ethics support services have been developed to support healthcare professionals in dealing with ethical dilemmas, including moral case deliberation. In this article, we describe how moral case deliberation works in daily practice, illustrated with a case example from the neurology ward. The article is meant as an introduction to moral case deliberation according to the dilemma method. We show its relevance to the clinic and the context needed to put it into practice.