Effects of HEMA on water evaporation from water-HEMA mixtures.
The aims of this research were: (1) to determine the relative rates of evaporation of water and HEMA, and (2) to determine the effects of increasing concentrations of HEMA on the rate of evaporation of water from water and HEMA mixtures. Ten microliters of each solution (100% H2O, 75% H2O-25% HEMA, 50% H2O-50% HEMA, 25% HEMA, 100% HEMA) were placed on the pan of a thermogravimetric analysis instrument held at 37 degrees C. The rate of spontaneous weight loss was measured as a function of time and relative humidity (RH) and compared statistically using ANOVA and Scheffé F test. The rate of evaporation of pure water was 32-fold higher than that of 100% HEMA. Addition of HEMA to water lowered the rate of evaporation of water from the water-HEMA mixtures in a manner that was proportional to its effect on lowering the vapor pressure of water (p < 0.05 comparing 50% HEMA with 75% HEMA). The rate of evaporation of water from water-HEMA mixtures was higher (p < 0.05) when the ambient gas was at 0% RH than when it was at 51% RH. The results indicate that as water evaporates from water-HEMA mixtures, the concentration of HEMA rises because it is relatively non-volatile. This rise in HEMA concentration lowers the vapor pressure of water making it more difficult to remove the last amounts of water. This residual water may interfere with polymerization of adhesive monomers, thereby lowering the quality of the hybrid layer.


MetaSV: an accurate and integrative structural-variant caller for next generation sequencing.
Structural variations (SVs) are large genomic rearrangements that vary significantly in size, making them challenging to detect with the relatively short reads from next-generation sequencing (NGS). Different SV detection methods have been developed; however, each is limited to specific kinds of SVs with varying accuracy and resolution. Previous works have attempted to combine different methods, but they still suffer from poor accuracy particularly for insertions. We propose MetaSV, an integrated SV caller which leverages multiple orthogonal SV signals for high accuracy and resolution. MetaSV proceeds by merging SVs from multiple tools for all types of SVs. It also analyzes soft-clipped reads from alignment to detect insertions accurately since existing tools underestimate insertion SVs. Local assembly in combination with dynamic programming is used to improve breakpoint resolution. Paired-end and coverage information is used to predict SV genotypes. Using simulation and experimental data, we demonstrate the effectiveness of MetaSV across various SV types and sizes. Code in Python is at http://bioinform.github.io/metasv/. firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Prometaphase forces towards opposite spindle poles are not independent: an on/off control system is identified by ultraviolet microbeam irradiations.
Individual spindle fibres in prometaphase spermatocytes of the cricket, Neocurtilla hexadactyla, were irradiated with an ultraviolet microbeam. The stretched heteromorphic bivalent (X2Y) contracted to about 75% of its pre-irradiation length after irradiation of either of its two oppositely directed spindle fibres (i.e., the X2 or Y spindle fibre). The X2Y bivalent also contracted after irradiation of the connection between the kinetochores of the univalent X1 chromosome and the Y chromosome but it did not contract after irradiation of autosomal spindle fibres or of the spindle fibre of the X1 univalent sex chromosome. The spindles sometimes shrank after irradiation, but contraction of the X2Y bivalent was independent of spindle shrinkage. The data strongly suggest that the oppositely directed forces on a bivalent are not independent. One reason is that the X2Y contractions were asymmetrical: during contraction one kinetochore remained fixed in position while the other kinetochore (facing the opposite pole) moved towards the equator. In 75% of the cases the stationary kinetochore was associated with the irradiated spindle fibre. Thus, we suggest that the irradiation of a spindle fibre produces a state analogous to rigor in the irradiated spindle fibre and, because of effects on the control system, produces relaxation of tension in the oppositely directed non-irradiated spindle fibre, so that the kinetochore associated with the non-irradiated spindle fibre moves towards the equator. These experiments have identified a control system that coordinates force production to opposite poles.


Reactive Vega: A Streaming Dataflow Architecture for Declarative Interactive Visualization.
We present Reactive Vega, a system architecture that provides the first robust and comprehensive treatment of declarative visual and interaction design for data visualization. Starting from a single declarative specification, Reactive Vega constructs a dataflow graph in which input data, scene graph elements, and interaction events are all treated as first-class streaming data sources. To support expressive interactive visualizations that may involve time-varying scalar, relational, or hierarchical data, Reactive Vega's dataflow graph can dynamically re-write itself at runtime by extending or pruning branches in a data-driven fashion. We discuss both compile- and run-time optimizations applied within Reactive Vega, and share the results of benchmark studies that indicate superior interactive performance to both D3 and the original, non-reactive Vega system.


Inverse sex effects on performance of domestic dogs (Canis familiaris) in a repeated problem-solving task.
The authors investigated differences between female and male pet dogs in physical cognition using an object manipulation task. Subjects (24 females and 23 males of different breeds) had to open a box in order to obtain a food reward during 3 consecutive trials, and latency times before success were measured. Males were significantly more successful in opening the box during the first trial. However, this sex difference was inversed when successful individuals were retested. During the following 2 trials, females were more successful than males, indicating that they were able to improve their skills more quickly once they had managed to succeed for a first time. Sex-specific dynamics in repeated problem-solving tasks might be an important contributor to individual differences in cognitive performance of pet dogs.


Your child is brain dead.
One to two per cent of admissions to Paediatric Intensive Care Units eventually fulfil the criteria for brain death, implying the need for very difficult decisions. Brain death is defined as irreversible loss of function of the whole brain. The diagnostic criteria caused a great deal of anxiety but are now the subject of a consensus approach. When the situation can be anticipated it is of immense value for the professional staff to develop a good working relationship with the parents to help and support them through the phase of impending disaster and facing the issue when the time actually comes. However, it is vital to help parents to make their own decision regarding continuation or otherwise of life support and they should be supported in whichever decision they take. They must be absolutely convinced that the child is brain dead and this territory may have to be covered again and again in discussion, questions must be answered factually, and time allowed for reflection. Stage management of the process of 'switching off' is vital and the parents' wishes may vary widely from one family to another. They must be warned what is likely to happen and provided with appropriate privacy and support for expression of their grief. The question of asking for post-mortem permission has to be handled sensitively and long-term support for the parents must be offered. Education of undergraduate and postgraduate doctors in this area is now receiving more attention with skills being increased by video teaching and role play. It should not be overlooked that the professional staff attending such patients sometimes require counselling and support themselves.


Computational design of nanoparticle drug delivery systems for selective targeting.
Ligand-functionalized nanoparticles capable of selectively binding to diseased versus healthy cell populations are attractive for improved efficacy of nanoparticle-based drug and gene therapies. However, nanoparticles functionalized with high affinity targeting ligands may lead to undesired off-target binding to healthy cells. In this work, Monte Carlo simulations were used to quantitatively determine net surface interactions, binding valency, and selectivity between targeted nanoparticles and cell surfaces. Dissociation constant, KD, and target membrane protein density, ρR, are explored over a range representative of healthy and cancerous cell surfaces. Our findings show highly selective binding to diseased cell surfaces can be achieved with multiple, weaker affinity targeting ligands that can be further optimized by varying the targeting ligand density, ρL. Using the approach developed in this work, nanomedicines can be optimally designed for exclusively targeting diseased cells and tissues.


Toward anisotropic materials via forced assembly coextrusion.
Multilayer coextrusion offers a diverse platform to examine layer dependent confinement effects on self-assembling nanomaterials via conventional extrusion technology. A triblock copolymer (BCP) with a cylindrical microstructure was processed via "forced assembly" to elucidate the effect of microdomain orientation on the mechanical behavior of multilayer films. The mechanical response was investigated in both the extrusion (ED) and transverse directions (TD) of the multilayer systems, revealing an influence of both cylinder-orientation and the interface on the mechanical response with decreasing layer thickness. The stress-strain curves for samples with the stress field along the cylinder axis revealed a sharp yielding phenomenon, while curves for specimens with the stress field applied perpendicular to the axis exhibited weak yielding behavior. The extensibility of the multilayer films stressed in the ED increases with decreasing layer thickness, but remains constant when deformed along the TD. Coextrusion technology allows for tunable mechanical toughness in industrial grade polymers via a continuous process. By altering the layer thickness of the two polymeric materials, we can tune the mechanics from strong, brittle behavior to a tough, ductile response by manipulation of the hierarchical structure. The enabling technology provides a unique platform to couple the inherent mechanical response of dissimilar polymers and allows for the design of composite materials with tailored mechanics.


Effect of surface mobility on the particle sliding along a bubble or a solid sphere.
The sliding velocity of glass beads on a spherical surface, made either of an air bubble or of a glass sphere held stationary, is measured to investigate the effect of surface mobility on the particle sliding velocity. The sliding process is recorded with a digital camera and analyzed frame by frame. The sliding glass bead was found to accelerate with increasing angular position on the collector's surface. It reaches a maximum velocity at an angular position of about 100 degrees and then, under certain conditions, the glass bead leaves the surface of the collector. The sliding velocity of the glass bead depends strongly on the surface mobility of a bubble, decreasing with decreasing surface mobility. By a mobile surface we mean one which cannot set up resistive forces to an applied stress on the surface. The sliding velocity on a rigid surface, such as a glass sphere, is much lower than that on a mobile bubble surface. The sliding velocity can be described through a modified Stokes equation. A numerical factor in the modified Stokes equation is determined by fitting the experimental data and is found to increase with decreasing surface mobility. Hydrophobic glass beads sliding on a hydrophobic glass sphere were found to stick at the point of impact without sliding if the initial angular position of the impact is less than some specific angle, which is defined as the critical sticking angle. The sticking of the glass beads can be attributed to the capillary contracting force created by the formation of a cavity due to spontaneous receding of the nonwetting liquid from the contact zone. The relationship between the critical sticking angle and the particle size is established based on the Yushchenko [J. Colloid Interface Sci. 96 (1983) 307] analysis.


Cell kinetic effects of incorporated 3H-thymidine on proliferating human lymphocytes: flow cytometric analysis using the DNA/nuclear protein method.
Phytohemagglutinin-stimulated human peripheral blood lymphocytes incorporating high concentrations of 3H-thymidine accumulate in G2 and show a consequent reduction in the number of cells entering M (division delay). The simultaneous flow cytometric analysis of DNA content (propidium iodide fluorescence) and nuclear protein content (fluorescein isothiocyanate fluorescence) allows for the accurate quantitation of these events; G2 and M are separated in the bivariate distributions. A good correlation was observed between mitotic indices, quantitated by manually counting mitotic cells, and integration of the M area in DNA/nuclear protein histograms. Moreover, significant differences in G2 nuclear protein levels were found between untreated and 3H-thymidine-treated lymphocytes. In order to characterize this effect, G2 was empirically divided into low nuclear protein (G2A) and high nuclear protein (G2B) compartments. 3H-thymidine caused an initial accumulation of lymphocytes in G2A, followed within 3-6 h by a gradual movement of some cells into G2B, with a subsequent accumulation of cells in G2B. The results suggest that the distribution of cells in G2 (G2A and G2B), the average nuclear protein content of G2B cells, and the proportion of cells in M are parameters that when used in combination provide a unique description of radiobiological effects.


New developments in the management of advanced melanoma - role of pembrolizumab.
Cancer immunotherapy is now recognized to be fundamental in modern oncology, because immune system recruitment may represent a powerful and innovative strategy in cancer therapy. Pembrolizumab, a highly selective humanized monoclonal antibody directly blocking the interaction between programmed cell death-1 expressed by tumor-associated T-cells and its ligand programmed cell death-L1 present on tumor and stromal cells, was recently approved by US Food and Drug Administration for the treatment of patients with unresectable or metastatic melanoma and disease progression upon ipilimumab and BRAF inhibitor. This review will focus on the clinical development and use of pembrolizumab in the clinical practice and in the management of advanced melanoma.


Unintended Pregnancy in Women Living with HIV in Sub-Saharan Africa: A Systematic Review and Meta-analysis.
In 2014, about 1.5 million pregnancies occurred among HIV-positive women in low and middle-income countries. To pool magnitude and factors associated with unintended pregnancy in women living with HIV in sub-Saharan Africa, a systematic search of electronic databases was undertaken in November 2016. Pooling the magnitude of unintended pregnancy reported by 14 studies yielded a crude summary prevalence of 55.9%. The magnitude of unwanted pregnancy and mistimed pregnancy in six studies ranged from 14 to 59 and 9 to 47.2%, respectively. Contraceptive failure was an important factor for many unintended pregnancies. The magnitude of unintended pregnancy was significantly higher in HIV-positive women than for HIV-negative women in three out of six studies. The available evidence suggests that there is a high magnitude of unintended pregnancy in this population. Improving effective contraceptive utilization is thus a priority to address unintended pregnancies and to prevent mother to child transmission of HIV. PROSPERO Number: CRD42016051310.


Pathogenesis of polycythemia vera.
Polycythemia vera (PV) is an acquired clonal myeloproliferative disorder characterized by increased production of mature red cells. We still lack a molecular target responsible for this disorder; however, recent investigations have focused on a number of molecules involved in signal transduction pathways mediated by erythropoietin (Epo) and other growth factors. Here we review the implication of these molecules in the pathogenesis of PV. The material reviewed in this work includes articles published in journals covered by Medline. We also include data obtained in our laboratory regarding to the significance of apoptosis inhibitory proteins in erythroid development. Overproduction of erythroid cells in PV is particular in that it occurs in the absence of a recognizable physiologic stimulus, since circulating serum levels of Epo are normal or lower than normal. Genetic analysis as well as in vitro studies, have established an essential role for Epo in the survival and maturation of committed erythroid progenitors. Epo initiates its cellular response by binding to the Epo receptor (EpoR) expressed on the surface of immature erythroblasts. Following ligand binding, EpoR is known to activate a cytoplasmic protein tyrosine kinase, Jak2 which triggers a signal transduction cascade that leads to the development of early erythroid progenitors into mature erythroblast cells. Although the mechanism underlying the increased erythroid production in PV is not well understood, a number of causes have recently came for which may provide insights not only for the pathogenesis of PV but also for a fundamental biological process: the mechanism whereby a multipotential stem cell gives rise to a particular cell lineage.


MTAP loss correlates with an immunosuppressive profile in GBM and its substrate MTA stimulates alternative macrophage polarization.
Glioblastoma (GBM) is a lethal brain cancer known for its potent immunosuppressive effects. Loss of Methylthioadenosine Phosphorylase (MTAP) expression, via gene deletion or epigenetic silencing, is one of the most common alterations in GBM. Here we show that MTAP loss in GBM cells is correlated with differential expression of immune regulatory genes. In silico analysis of gene expression profiles in GBM samples revealed that low MTAP expression is correlated with an increased proportion of M2 macrophages. Using in vitro macrophage models, we found that methylthioadenosine (MTA), the metabolite that accumulates as a result of MTAP loss in GBM cells, promotes the immunosuppressive alternative activation (M2) of macrophages. We show that this effect of MTA on macrophages is independent of IL4/IL3 signaling, is mediated by the adenosine A2B receptor, and can be pharmacologically reversed. This study suggests that MTAP loss in GBM cells may contribute to the immunosuppressive tumor microenvironment, and that MTAP status should be considered for characterizing GBM immune states and devising immunotherapy-based approaches for treating MTAP-null GBM.


Role of Ape1 and base excision repair in the radioresponse and heat-radiosensitization of HeLa Cells.
The mechanism by which heat sensitizes mammalian cells to ionizing radiation remains to be elucidated. We determined whether base excision repair (BER) is involved in heat-radiosensitization and report novel findings that provide insight regarding the role of BER in the radiation response of HeLa cells. An siRNA approach was utilized to suppress expression of AP endonuclease (Ape1), a critical enzyme of BER. Clonogenic survival curves were obtained for HeLa cells expressing normal or reduced Ape1 content and which had been irradiated, and these were compared to survival curves from cells that were irradiated prior to hyperthermia treatment. The amount of heat-radiosensitization observed in Ape1-suppressed cells was similar to or slightly greater than that observed in cells expressing near-normal levels of Ape1. Interestingly, we also found that for unheated HeLa cells, suppressed expression of Ape1 resulted in enhanced resistance to X-rays. The data suggest that Ape1, and therefore BER, is not involved in heat-radiosensitization. However, the observation that suppressed expression of Ape1 results in enhanced radioresistance supports the notion that BER may be detrimental to the survival of irradiated cells.


An exploration of socioeconomic variation in lifestyle factors and adiposity in the Ontario Food Survey through structural equation modeling.
Socioeconomic indicators have been inversely associated with overweight and obesity, with stronger associations observed among women. The objective of the present secondary analysis was to examine the relationships among socioeconomic measures and adiposity for men and women participating in the Ontario Food Survey (OFS), and to explore lifestyle factors as potential mediators of these associations. The cross-sectional 1997/98 OFS collected anthropometric measurements, a food frequency questionnaire, data on socio-demographics (age, sex, income, and education) and physical activity from 620 women and 467 men, ages 18 to 75. Based on the 2003 Health Canada guidelines, waist circumference and BMI values were used to derive least risk, increased risk, and high risk adiposity groups. Structural equation modeling was conducted to examine increased risk and high risk adiposity in relation to education and income, with leisure time physical activity, fruit and vegetable intake, and smoking status included as potential mediators of these associations. The probability of high risk adiposity was directly associated with education (beta-0.19, p < 0.05) and income (beta-0.22, p < 0.05) for women, but not for men. Fruit and vegetable intake was a marginally significant mediator of the relationship between education and high risk adiposity for women. Increased risk adiposity was not associated with income or education for men or women. The socioeconomic context of adiposity continues to differ greatly between men and women. For women only in the OFS, fruit and vegetable intake contributed to the inverse association between education and high risk adiposity; however, additional explanatory factors are yet to be determined.


Effects of chronic mild stress on serum complement activity, saccharin preference, and corticosterone levels in Flinders lines of rats.
Complement proteins and fragments participate in the induction and modulation of specific and nonspecific immune reactions. We have examined the effect of 4 weeks of chronic mild stress (CMS) on complement sheep red blood cell hemolytic activity measured in CH50 units in two selectively bred lines of rats, the Flinders resistant line (FRL) and the Flinders sensitive line (FSL), that differ in cholinergic sensitivity and behavioral characteristics. Additionally, CMS-induced hedonic deficit (decreased preference for 0.02% saccharin over water) and serum corticosterone levels were compared in FRL and FSL rats. CMS caused a significantly (p < 0.01) greater decline in CH50 responses in FSL (-15%) than in FRL (-7%) rats. This was accompanied by a significant (p < 0.01) suppression of saccharin preference over a 24 h period in both FRL and FSL rats. Both lines showed a similar, more than 2-fold (p < 0.01) increase in corticosterone levels following CMS. These results further confirm that CMS induces a depressive-like state in rats as well as the validity of the FSL rat as a genetic model of depression. They also indicate that the effect of stress on the immune system can be monitored by measuring the complement CH50 response.


On the structural connectivity of large-scale models of brain networks at cellular level.
The brain's structural connectivity plays a fundamental role in determining how neuron networks generate, process, and transfer information within and between brain regions. The underlying mechanisms are extremely difficult to study experimentally and, in many cases, large-scale model networks are of great help. However, the implementation of these models relies on experimental findings that are often sparse and limited. Their predicting power ultimately depends on how closely a model's connectivity represents the real system. Here we argue that the data-driven probabilistic rules, widely used to build neuronal network models, may not be appropriate to represent the dynamics of the corresponding biological system. To solve this problem, we propose to use a new mathematical framework able to use sparse and limited experimental data to quantitatively reproduce the structural connectivity of biological brain networks at cellular level.


A TGF-β/KLF10 signaling axis regulates atrophy-associated genes to induce muscle wasting in pancreatic cancer.
Cancer cachexia, and its associated complications, represent a large and currently untreatable roadblock to effective cancer management. Many potential therapies have been proposed and tested-including appetite stimulants, targeted cytokine blockers, and nutritional supplementation-yet highly effective therapies are lacking. Innovative approaches to treating cancer cachexia are needed. Members of the Kruppel-like factor (KLF) family play wide-ranging and important roles in the development, maintenance, and metabolism of skeletal muscle. Within the KLF family, we identified KLF10 upregulation in a multitude of wasting contexts-including in pancreatic, lung, and colon cancer mouse models as well as in human patients. We subsequently interrogated loss-of-function of KLF10 as a potential strategy to mitigate cancer associated muscle wasting. In vivo studies leveraging orthotopic implantation of pancreas cancer cells into wild-type and KLF10 KO mice revealed significant preservation of lean mass and robust suppression of pro-atrophy muscle-specific ubiquitin ligases Trim63 and Fbxo32, as well as other factors implicated in atrophy, calcium signaling, and autophagy. Bioinformatics analyses identified Transforming growth factor beta (TGF-β), a known inducer of KLF10 and cachexia promoting factor, as a key upstream regulator of KLF10. We provide direct in vivo evidence that KLF10 KO mice are resistant to the atrophic effects of TGF-β. ChIP-based binding studies demonstrated direct binding to Trim63, a known wasting-associated atrogene. Taken together, we report a critical role for the TGF-β/KLF10 axis in the etiology of pancreatic cancer-associated muscle wasting and highlight the utility of targeting KLF10 as a strategy to prevent muscle wasting and limit cancer-associated cachexia.


The neural correlates of 'vitality form' recognition: an fMRI study: this work is dedicated to Daniel Stern, whose immeasurable contribution to science has inspired our research.
The observation of goal-directed actions performed by another individual allows one to understand what that individual is doing and why he/she is doing it. Important information about others' behaviour is also carried out by the dynamics of the observed action. Action dynamics characterize the 'vitality form' of an action describing the cognitive and affective relation between the performing agent and the action recipient. Here, using the fMRI technique, we assessed the neural correlates of vitality form recognition presenting participants with videos showing two actors executing actions with different vitality forms: energetic and gentle. The participants viewed the actions in two tasks. In one task (what), they had to focus on the goal of the presented action; in the other task (how), they had to focus on the vitality form. For both tasks, activations were found in the action observation/execution circuit. Most interestingly, the contrast how vs what revealed activation in right dorso-central insula, highlighting the involvement, in the recognition of vitality form, of an anatomical region connecting somatosensory areas with the medial temporal region and, in particular, with the hippocampus. This somatosensory-insular-limbic circuit could underlie the observers' capacity to understand the vitality forms conveyed by the observed action.


[Emergency dialysis at Haukeland hospital in 1999].
The incidence of acute renal failure requiring dialysis is not known in our country. The criteria for acute dialysis are not uniformly accepted, neither is there consensus on dialysis strategy in critically ill patients. We describe the acute dialysis activity in our hospital in 1999. We have retrospectively recorded the indications for dialysis and the course and hospital mortality in all patients treated with acute dialysis in 1999. 108 patients were treated with 670 dialysis procedures; the incidence was 20.5 per 100,000 inhabitants. Continuous veno-venous haemodiafiltration accounted for 37% of all treatments. In 50% of the patients acute renal failure occurred after surgery or serious infections with a mortality of 57% and 62% respectively. Total dialysis mortality was 45%. The mortality in patients with malignancy and peripheral vascular disease was 90% and 75% respectively. 10% of the patients needed chronic dialysis. The incidence of acute dialysis was higher than previously reported from European countries and may be related to the general increase in active treatment of patients with complicated diseases. The mortality rates are persistently high. Close cooperation between nephrologists and intensivists in the treatment of these patients is essential.


Herpes simplex virus induces extensive modification and dynamic relocalisation of the nuclear mitotic apparatus (NuMA) protein in interphase cells.
The nuclear mitotic apparatus (NuMA) protein is a component of the nuclear matrix in interphase cells and an essential protein for the formation of mitotic spindle poles. We used herpes simplex virus (HSV), an enveloped DNA virus that replicates in the nucleus, to study the intra-nuclear dynamics of NuMA in infected cells. This study shows that NuMA is extensively modified following HSV infection, including phosphorylation of an unidentified site(s), and that it depends to an extent on viral DNA synthesis. Although NuMA is insoluble in uninfected interphase cells, HSV infection induced solubilisation and dynamic relocalisation of NuMA, whereupon the protein became excluded from viral replication compartments -- sites of virus transcription and replication. Live cell, confocal imaging showed that NuMA localisation dramatically changed from the early stages (diffusely nuclear, excluding nucleoli) to late stages of infection (central diminuition, but remaining near the inner nuclear peripheries). In addition, NuMA knockdown using siRNA suggested that NuMA is important for efficient viral growth. In summary, we suggest that NuMA is required for efficient HSV infection, and identify further areas of research that address how the virus challenges host cell barriers.


Impact of comorbidities at diagnosis on prostate cancer treatment and survival.
The aim of this study was to assess the associations of comorbidities with primary treatment of prostate cancer (PCa) patients and of comorbidities with PCa-specific mortality (PCSM) compared to other-cause mortality (OCM) in Switzerland. We included 1527 men diagnosed with PCa in 2000 and 2001 in the canton of Zurich. Multiple imputation methods were applied to missing data for stage, grade and comorbidities. Multinomial logistic regression analyses were used to explore the associations of comorbidities with treatment. Cox regression models were used to estimate all-cause mortality, and Fine and Gray competing risk regression models to estimate sub-distribution hazard ratios for the outcomes PCSM and OCM. Increasing age was associated with a decreasing probability of receiving curative treatment, whereas an increasing Charlson Comorbidity Index (CCI) did not influence the treatment decision as strongly as age. The probability of OCM was higher for patients with comorbidities compared to those without comorbidities [CCI 1: hazard ratio 2.07 (95% confidence interval 1.51-2.85), CCI 2+: 2.34 (1.59-3.44)]; this was not observed for PCSM [CCI 1: 0.79 (0.50-1.23), CCI 2+: 0.97 (0.59-1.59)]. In addition, comorbidities had a greater impact on the patients' mortality than age. The results of the current study suggest that chronological age is a stronger predictor of treatment choices than comorbidities, although comorbidities have a larger influence on patients' mortality. Hence, inclusion of comorbidities in treatment choices may provide more appropriate treatment for PCa patients to counteract over- or undertreatment.


Protective Effect of Selenium Against Cisplatin-Induced Ototoxicity in an Experimental Design.
Cisplatin is an effective chemotherapeutic agent in the treatment of several types of malignant solid tumors but its clinical use is associated with ototoxicity. In the present study, we investigated the effect of selenium administration on lipid peroxidation (malondialdehyde [MDA]) and cisplatin-induced ototoxicity in rats. Healthy wistar albino rats (n = 21) were randomly divided into 3 groups: control (C), cisplatin (Cis), cisplatin and selenium (Cis+Se). Cisplatin was administered for 3 days to Cis and Cis+Se groups. Cis+Se group received selenium 5 days before cisplatin injection and continued for 11 consecutive days. Hearing thresholds and lipid peroxidation (MDA) levels of the rats were recorded before injections and at the end of experimental protocol. The cochleas of animals were harvested for histologic and immunuhistochemical examinations. In biochemichal analyses, pretreatment with selenium prevented the elevation of MDA levels in Cis+Se group rats. Moreover, animals in Cis+Se group had better hearing threshold levels than animals in cis group. Samples obtained from the animals in Cis group revealed extensive loss of the normal microarchitecture of the organ of Corti. On the other hand, animals in Cis+Se group exhibited a preservation of the morphology of the organ of Corti and outer hair cells. In the immunohistochemical examinations of cochlear tissues stained with anti-caspase-3, a higher degree of immunopositivity was found in the Cis group. When Cis+Se group and Cis group were compared, significantly less immunopositivity occurred in the Cis+Se group (P < 0.05). Thus, it appears that pretreatment with selenium may reduce cisplatin-induced ototoxicity in rats.


Recurrent infection with genetically identical pneumococcal isolates in a patient with interleukin-1 receptor-associated kinase-4 deficiency.
Interleukin-1 receptor-associated kinase (IRAK)-4 deficiency is a rare primary immunodeficiency disorder characterized by severe, invasive infections with Streptococcus pneumoniae. Using the PFGE technique a genetic linkage was found between two S. pneumoniae serotype 14 isolates causing arthritis and meningitis at 3 and 5(1/2) years of age, respectively, in a boy with IRAK-4 deficiency. This finding suggested that patients with IRAK-4 deficiency may harbour persistent strains of pneumococci. Alternatively, reinfection with strains from close contacts of the patient might cause recurrent invasive disease. It is proposed that eradication of pneumococci from the nasopharynx, and immunization of household contacts may prevent recurrent infection in IRAK-4-deficient patients.


Structural basis for epitope sharing between group 1 allergens of cedar pollen.
The group 1 allergens are a major cause of cedar pollen hypersensitivity in several geographic areas. Allergens from several taxa have been shown to cross-react. The goal of these studies was to compare the structural features of the shared and unique epitopes of the group 1 allergen from mountain cedar (Jun a 1) and Japanese cedar (Cry j 1). An array of overlapping peptides from the sequence of Jun a 1 and a panel of monoclonal anti-Cry j 1 antibodies were used to identify the IgE epitopes recognized by cedar-sensitive patients from Texas and Japan. IgE from Japanese patients reacted with peptides representing one of the two linear epitopes within the highly conserved beta-helical core structure and both epitopes within less ordered loops and turns near the N- and C-termini of Jun a 1. A three-dimensional (3D) model of the Cry j 1, based on the crystal structure of Jun a 1, indicated a similar surface exposure for the four described epitopes of Jun a 1 and the homologous regions of Cry j 1. The monoclonal antibodies identified another shared epitope, which is most likely conformational and a unique Cry j 1 epitope that may be the previously recognized glycopeptide IgE epitope. Defining the structural basis for shared and unique epitopes will help to identify critical features of IgE epitopes that can be used to develop mimotopes or identify allergen homologues for vaccine development.


Three-dimensional sonographic morphologic assessment in complex adnexal masses: preliminary experience.
To evaluate the role of three-dimensional transvaginal sonography in assessing complex adnexal masses. Forty-one women (mean age, 49.5 years; range, 23-75 years) with the diagnosis of complex adnexal masses on the basis of two-dimensional transvaginal sonography were reevaluated by three-dimensional transvaginal sonography. Two different sonologists evaluated the two- and three-dimensional transvaginal sonograms. Criteria indicative of malignancy included the presence of gross papillary projections, solid areas, and solid echogenicity for both techniques. Three women (7%) had bilateral masses, giving a total of 44 masses that were ultimately assessed. A definitive histologic diagnosis was obtained in every case after surgical tumor removal. Twenty-one tumors (47.7%) were proved malignant, and 23 (52.3%) were benign. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for two- and three-dimensional transvaginal sonography were 90%, 61%, 68%, 87%, and 75% and 100%, 78%, 81%, 100%, and 89%, respectively. There were no statistical differences between two- and three-dimensional transvaginal sonography (McNemar test, P = .687). The agreement between both examiners was high (kappa index = 0.71). The use of three-dimensional transvaginal sonography does not significantly improve the two-dimensional transvaginal sonographic morphologic assessment of complex adnexal masses; however, we found it useful for reinforcing initial diagnostic impressions.


Optimality principles and flow orderliness at the branching points of cerebral arteries.
The cerebral arteries present an optimum blood flow/vessel radius relation. However, branch angles may vary widely in the cerebral arteries because the parametric optimization of branch angles is irrelevant in terms of energy cost. The position of the flow divider in extracranial arteries has been suggested to be optimum in flow orderliness. No data exist on the flow divider of cerebral arteries. Thus, we hypothesized that in the cerebral arteries the apex of the bifurcations, which is known to be the site of maximum hemodynamic stress in a vascular network, may normally lie in a non-optimum position relative to the dividing flow streamline in the parent vessel, leading to disturbed laminar flow and increased vessel wall shear stress at the apical region despite the optimum blood flow/vessel radius relation. The objective of this study was to test our hypothesis. We measured the branch angles and diameters of parent and branch segments of the anterior cerebral artery system from lateral projections to minimize the measurement error on angiographs chosen at random from normal sets. The position of the apex of the bifurcations in relation to the ostium of the parent artery (gamma) and the ratio of the branch diameters (d2/d1) were compared. Optimum curves for these parameters were calculated by a mathematical model. In addition, the separation of flow streamlines according to gamma was calculated for each bifurcation and related to the division of flow required by each branch according to the optimum blood flow/vessel radius relation. The data points on gamma and d2/d1 and the separation of flow according to gamma and the division of flow required by the branches were found to scatter around the optimum curves. However, a trend toward the theoretical optimum is discernible. The data points are suggested to be a random sample from a normal distribution around the optimum (.40 < P < .50). The bifurcations of the cerebral arteries appear to be optimized to avoid increased hemodynamic stresses both globally and locally in the same manner as extracranial arteries.


The frequency of circulating integrin α4β7+ cells correlates with protection against Helicobacter pylori infection in immunized mice.
Chronic Helicobacter pylori infection is the cause of peptic ulcers in a subpopulation of individuals and a risk factor for the development of gastric cancer. A vaccine against H pylori infection can prevent the acquisition of the infection and protect against reinfections. Clinical trials to date evaluating the efficacy of H pylori vaccines in human challenge models have shown moderate to poor protection with difficulties in predicting efficacy. Thus, while further studies are needed to design an effective vaccine, we also need to find relevant correlates for vaccine efficacy. To find immune correlates to vaccine efficacy, the frequencies of neutrophils, eosinophils and inflammatory monocytes and CD4+ T-cell memory and mucosa homing integrin α4β7+ cells were assessed by flow cytometry in the blood of mice after vaccination. H pylori antigens and cholera toxin or the multiple mutant CT (mmCT) were administered via the sublingual (SL) and intragastric route (IG). The vaccinated mice were infected with H pylori strain SS1 bacteria, and colonization in the stomach and immune responses were evaluated. The H pylori vaccine was effective in reducing bacterial load in the stomach of mice and enhancing immune responses compared to unvaccinated infection controls. In the blood of mice after SL or IG route of vaccination, we observed changes in frequencies of innate and adaptive immune cell subsets compared to infection controls. Remarkably, the frequency of circulating mucosal homing α4β7+ CD4+ T cells after vaccination correlated with low bacterial load in the stomach of individual mice irrespective of the immunization route. Our study shows that the innate and adaptive immune cell subsets can be measured in the blood after vaccination and that increased frequency of α4β7+ CD4+ in the blood after immunization could be used as a predictive marker for the efficacy of vaccine against H pylori infection.


On the states of stress and strain adjacent to a crack in a strain-limiting viscoelastic body.
The viscoelastic Kelvin-Voigt model is considered within the context of quasi-static deformations and generalized with respect to a nonlinear constitutive response within the framework of limiting small strain. We consider a solid possessing a crack subject to stress-free faces. The corresponding class of problems for strain-limiting nonlinear viscoelastic bodies with cracks is considered within a generalized formulation stated as variational equations and inequalities. Its generalized solution, relying on the space of bounded measures, is proved rigorously with the help of an elliptic regularization and a fixed-point argument.


Discussion on gemcitabine combined with targeted drugs in the treatment of pancreatic cancer.
Pancreatic cancer is a malignant tumor with poor prognosis. The treatment of pancreatic cancer depends on the tumor stage and type, and includes local treatment (surgery, radiotherapy and ablation intervention) and systemic therapy (chemotherapy, targeted therapy and immunotherapy). We read with great interest the review "Effective combinations of anti-cancer and targeted drugs for pancreatic cancer treatment" published on World J Gastroenterol and intended to share some of our perspectives in pancreatic cancer treatment. This review presents the therapeutic effects of the combination of gemcitabine and targeted drugs, which gives us a deeper insight into the combination treatments for pancreatic cancer.


Therapeutic effects of substances occurring in higher Basidiomycetes mushrooms: a modern perspective.
This review highlights some of the recently isolated and identified substances of higher Basidiomycetes mushrooms origin that express promising antitumor, immune modulating, cardiovascular and hypercholesterolemia, antiviral, antibacterial, and antiparasitic effects. Medicinal mushrooms have a long history of use in folk medicine. In particular, mushrooms useful against cancers of the stomach, esophagus, lungs, etc. are known in China, Russia, Japan, Korea, as well as the U.S.A. and Canada. There are about 200 species of mushrooms that have been found to markedly inhibit the growth of different kinds of tumors. Searching for new antitumor and other medicinal substances from mushrooms and to study the medicinal value of these mushrooms have become a matter of great significance. However, most of the mushroom origin antitumor substances have not been clearly defined. Several antitumor polysaccharides such as hetero-beta-glucans and their protein complexes (e.g., xyloglucans and acidic beta-glucan-containing uronic acid), as well as dietary fibers, lectins, and terpenoids have been isolated from medicinal mushrooms. In Japan, Russia, China, and the U.S.A. several different polysaccharide antitumor agents have been developed from the fruiting body, mycelia, and culture medium of various medicinal mushrooms (Lentinus edodes, Ganoderma lucidum, Schizophyllum commune, Trametes versicolor, Inonotus obliquus, and Flammulina velutipes). Both cellular components and secondary metabolites of a large number of mushrooms have been shown to effect the immune system of the host and therefore could be used to treat a variety of disease states.


Nanoparticle Systems for Cancer Phototherapy: An Overview.
Photodynamic therapy (PDT) and photothermal therapy (PTT) are photo-mediated treatments with different mechanisms of action that can be addressed for cancer treatment. Both phototherapies are highly successful and barely or non-invasive types of treatment that have gained attention in the past few years. The death of cancer cells because of the application of these therapies is caused by the formation of reactive oxygen species, that leads to oxidative stress for the case of photodynamic therapy and the generation of heat for the case of photothermal therapies. The advancement of nanotechnology allowed significant benefit to these therapies using nanoparticles, allowing both tuning of the process and an increase of effectiveness. The encapsulation of drugs, development of the most different organic and inorganic nanoparticles as well as the possibility of surfaces' functionalization are some strategies used to combine phototherapy and nanotechnology, with the aim of an effective treatment with minimal side effects. This article presents an overview on the use of nanostructures in association with phototherapy, in the view of cancer treatment.


Nitrate reductase is responsible for elicitin-induced nitric oxide production in Nicotiana benthamiana.
Recent works have established a key role for nitric oxide (NO) in activating disease resistance in plants. Nitrate reductase (NR) is one of the enzymes that are capable of producing NO in plants. In a previous study, we reported that pathogen signals induce expression of NR genes in potato, suggesting the involvement of NR in NO production induced by pathogen signals. In this study, we cloned NR genes from Nicotiana benthamiana and investigated their involvement in NO production induced by INF1, a major elicitin secreted by Phytophthora infestans. Treatment of protoplasts prepared from N. benthamiana leaves with INF1 elevated NO production to a maximum level 1-3 h after treatment. INF1-induced NO generation was suppressed completely by an NO-specific scavenger, but partially by a nitric oxide synthase inhibitor. To investigate the involvement of NR in INF1-induced NO production, NR genes were silenced by virus-induced gene silencing. The NR-silenced plants showed yellowish leaves which resemble the characteristic of Arabidopsis NR double mutants. Silencing of NR genes significantly decreased both NO(2) (-)-producing activity and INF1-induced NO production, indicating that NR is involved in INF1-induced NO production. In contrast, overexpression of NbNR1 encoding N. benthamiana NR by Agrobacterium-mediated transient expression elevated NO(2) (-)-producing activity nine times over the control; however, INF1-induced NO production in protoplasts overexpressing NbNR1 was comparable with that in control protoplasts. These results suggest that NR is involved in INF1-induced NO production, and post-translational modification of NR or availability of substrate NO(2) (-) may be a rate-limiting step of NO production by NR.


Looking back--a history of progress: Society of Quality Assurance.
The Society of Quality Assurance (SQA) Historical Committee has tracked the impressive evolution of our society from the initial sharing of ideas at the 1980 Quality Assurance Roundtable meeting to the formation of the national SQA in 1984, to a 12-fold increase in membership numbers through 1993. This article describes annual meeting locations and attendance, membership numbers, and organizational information, as well as significant milestones for both the Quality Assurance Roundtable group and the SQA.


Amphiphilic redox-sensitive NIR BODIPY nanoparticles for dual-mode imaging and photothermal therapy.
Theranostics, integrating tumor treatment and diagnosis concurrently, has become an emerging and meaningful strategy in cancer therapy. In this work, an amphiphilic redox-sensitive near-infrared (NIR) BODIPY dye, which could be formed into nanoparticles (PEG-SS-BDP NPs) by self-assembly, was synthesized and possessed good capability of photothermal therapy (PTT), near-infrared fluorescence (NIRF) imaging, photoacoustic (PA) imaging and drug loading. The stable nanoparticles could be dissociated to turn on NIRF due to the rift of embedded disulfide bonds by glutathione (GSH). The enhanced fluorescence in vitro could be observed via confocal laser scanning microscopy (CLSM) after adding GSH, confirming the redox-sensitivity of disulfide bonds. NIRF and PA imaging demonstrated active accumulation in tumor and good imaging effect. At last, PEG-SS-BDP NPs could significantly suppress tumor growth in vivo upon irradiation. The amphiphilic redox-sensitive BODIPY nanoparticles provide a promising design strategy to formulate multifunctional stimuli-responsive theranostic nanoplatforms.


Myopathy and paraproteinemia with serum IgM binding to a high-molecular-weight muscle fiber surface protein.
We evaluated a 69-year-old man with Waldenström's macroglobulinemia (IgM-kappa M-protein) and progressive weakness over 2 to 3 years. The neurological examination showed symmetrical, predominantly proximal weakness. Electrophysiological testing revealed small brief motor unit potentials with fibrillations and positive sharp waves consistent with an irritative myopathy. The muscle biopsy specimen showed myopathic changes including variation in fiber size and increased connective tissue but no inflammation. IgM-kappa but not IgM-lambda was deposited along the surface of muscle fiber membranes. Serum IgM-kappa but not IgM-lambda from the patient stained the surface of normal human muscle fibers but not other regions of muscle fibers or other tissues. The serum IgM-kappa at dilutions up to 1:512,000 bound to a high-molecular-weight muscle protein by Western and dot blot studies. By enzyme-linked immunosorbent assay and Western blot analysis, serum IgM-kappa bound specifically to the muscle protein and not to other muscle or neural antigens, including GM1 ganglioside, myelin-associated glycoprotein, and sulfatide. We conclude that the myopathy in our patient is probably related to the presence of serum IgM-kappa antibodies directed against a muscle surface antigen. Characterization of the target antigen, a high-molecular-weight protein located specifically in muscle, should further elucidate the pathogenesis of this presumably humorally mediated immune myopathy.


Prospective Randomized Trial of Early Postoperative Enteral and Total Parenteral Nutrition for Treating Esophageal Cancer.
Surgical stress significantly decreases serum diamine oxidase (DAO), a marker of intestinal mucosal maturation and integrity. This study aimed to determine the effects of postoperative enteral and total parenteral nutrition (EN and TPN, respectively) in patients with esophageal cancer. This prospective randomized trial compared serum DAO activities, nutritional states, trace elements and complications between patients who underwent esophagectomy and received EN or TPN for seven days thereafter. Fifty-one patients were randomized to receive EN or TPN. The rates of change in serum DAO activity from the day before surgery were 0.79, 0.89 and 0.91 on postoperative days (POD) 1, 3 and 7, respectively, in the EN group, and 0.64, 0.76 and 1.06, respectively, in the TPN group, with no significant differences. Furthermore, the rates of changes in indicators of nutritional status, namely total protein, albumin, total cholesterol, trace element concentrations and infectious and non-infectious complications did not significantly differ between the groups. EN and/or TPN can be administered for early nutritional management until resumption of oral intake after esophagectomy according to the postoperative status of individual patients with esophageal cancer.


The selectivity of action of methotrexate in combination with 5-fluorouracil in xenografts of human colon adenocarcinomas.
This study was designed to examine the possibilities for increasing the therapeutic index in the treatment of human colon adenocarcinomas maintained as xenografts in immune-deprived mice using combinations of methotrexate (MTX) that preceded 5-fluorouracil (FUra). MTX, at a dose level of 100 mg/kg, increased the 5-phosphoribosyl-1-pyrophosphate (PRPP) concentration in three colon xenograft lines to a maximum between 14 and 24 hr after treatment. In murine bone marrow, concentrations of PRPP decreased progressively after MTX treatment, but in ileum there was a dramatic increase such that by 4 hr PRPP was 968% of control. The metabolism of [6-3H]FUra administered 24 hr after MTX was increased in ileum and resulted in an increased rate and a greater level of incorporation of [6-3H]FUra into RNA. Only a slight elevation in the incorporation of [6-3H]FUra into the RNA of one tumor line (HxELC2) was observed. The scheduling of FUra at a dose level of 25 mg/kg 24 hr after a priming dose of MTX (100 mg/kg) was at least as toxic as 100 mg of FUra per kilogram, administered alone. The dose-limiting toxicity was related to gastrointestinal damage; no bone marrow toxicity was detected. At dose levels of MTX up to 100 mg/kg, the increase in PRPP obtained in gastrointestinal tissue was greater than that observed in human colon xenografts 24 hr after treatment. A basis for increasing the therapeutic efficacy of FUra through a selective increase in tumor PRPP using MTX was not obtained in these studies.


Chemical immobilisation of giraffe to facilitate short procedures.
To describe the chemical immobilisation of giraffe (Giraffa camelopardalis) for short procedures at Taronga Zoo. A clinical report. Five giraffe. The weight of each animal was determined and pre-immobilisation fasting, careful preparation of the site, and planing were carried out prior to each procedure. Etorphine and acetylpromazine were used as the sole immobilising agents. During immobilisation, elevation of the head and constant monitoring of the heart rate, respiratory rate and temperature were employed; blood pressure and blood oxygen saturation (SpO2) were monitored in one case 5. Intravenous fluids were administered. None of the animals was intubated, and supplemental oxygen was administered via an intranasal cannula in case 5. At the end of the procedures, diprenorphine was administered to reverse immobilisation. Induction times were 8 to 15 min. The mean heart rate, respiratory rate and rectal temperature were 58.6 beats per min, 12.2 breaths per min and 37.1 degrees C, respectively. SpO2 in case 5 increased to over 90% with the administration of supplemental oxygen. Recovery to standing after reversal of immobilisation was rapid (2 or 3 min) in three cases, but took 30 and 31 min in the other two. Successful immobilisation of giraffe relies on consideration of all variables prior to and during the procedure. Careful selection and preparation of the venue and presence of sufficient numbers of adequately trained personnel are essential. Body weight determination allows accurate calculation of immunobilising agent dose. Pre-immobilisation fasting followed by elevation of head and neck help preclude regurgitation and aspiration. Relevant vital signs monitoring, fluid replacement and oxygen supplementation promote cardiopulmonary homeostasis. Preoperative planning to ensure efficient performance of the procedure helps minimise recumbency time and increase the likelihood of a successful outcome.


Getting and keeping Medicare certification. A quality assurance program for ambulatory surgical centers.
Getting and keeping Medicare certification is becoming more challenging every year. It is imperative that the ASC develop and maintain comprehensive documentation to verify compliance with the standards. The ASC should also plan for increased interaction with their state's PRO in the areas of medical necessity and appropriateness of care. Through proper design and implementation, the QA program should minimize the exposures to Medicare.


Use of metamodels for rapid discovery of narrow bandgap oxide photocatalysts.
New photocatalysts are traditionally identified through trial-and-error methods. Machine learning has shown considerable promise for improving the efficiency of photocatalyst discovery from a large potential pool. Here, we describe a multi-step, target-driven consensus method using a stacking meta-learning algorithm that robustly predicts bandgaps and H2 evolution activities of photocatalysts. Trained on small datasets, these models can rapidly screen a large space (>10 million materials) to identify promising, non-toxic compounds as candidate water splitting photocatalysts. Two effective compounds and two controls possessing optimal bandgap values (∼2 eV) but not photoactivity as predicted by the models were synthesized. Their experimentally measured bandgaps and H2 evolution activities were consistent with the predictions. Conspicuously, the two compounds with strong photoactivities under UV and visible light are promising visible-light-driven water splitting photocatalysts. This study demonstrates the power of machine learning and the potential of big data to accelerate discovery of next-generation photocatalysts.


Clinical usefulness of FDG-PET/CT for identification of abnormal extra-cardiac foci in patients with infective endocarditis.
The aim of the study was to evaluate the ability to detect extra-cardiac foci by means of whole-body 18F-fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) in patients with definite endocarditis (IE) according to the modified Duke criteria and investigate the clinical impact of the findings. From January 2011 to December 2015 we included 178 patients (mean age 66 ± 14 years, 25% female) with IE in this multicentre study. FDG-PET/CT was part of the work-up for extra-cardiac foci in the including hospitals and was performed at a median of 9 days (IQR 10) after IE was diagnosed. In 114 patients FDG-PET/CT identified 166 lesions: 52 (31%) infectious lesions, 21 (13%) cases of cancer, 7 (4%) cases of embolism, 60 (36%) reactive findings, and 26 (16%) other types of lesions. A total of 74 new extra-cardiac findings, not previously discovered by other modalities, were identified in 62 patients and resulted in additional investigations in 29 patients and a change in treatment in 18 patients (10%). The most frequent diagnoses discovered by FDG-PET/CT were colon polyps, cancer, and spondylodiscitis. There was a higher rate of findings leading to a change in treatment in patients above 67 years of age infected with other bacterial aetiologies than streptococci. FDG-PET/CT was useful to detect extra-cardiac foci. FDG-PET/CT findings may lead to unnecessary investigations. One out of 10 the patients with definite endocarditis had underwent a change in treatment regimen based on the FDG-PET/CT findings.


APE1 Upregulates MMP-14 via Redox-Sensitive ARF6-Mediated Recycling to Promote Cell Invasion of Esophageal Adenocarcinoma.
Esophageal adenocarcinoma (EAC) is an aggressive malignancy with poor clinical outcome. The incidence of EAC has been rising rapidly in the past three decades. Here, we showed that apurinic/apyrimidinic endonuclease (APE1) is overexpressed in EAC cell lines, and patients' samples of dysplasia and EAC. Downregulation of APE1 or inhibition of its redox function significantly repressed invasion. Overexpression of a redox-defective mutant, C65A, abrogated the proinvasive phenotype of APE1. APE1 regulated invasion via upregulation of matrix metalloproteinase 14 (MMP-14), which subsequently activated MMP-2, leading to degradation of the extracellular matrix in a redox-dependent manner. Downregulation of APE1 or inhibition of its redox function decreased the rate of endocytosis and recycling of MMP-14 protein. APE1 interacted with ARF6, a key regulator of MMP-14 recycling, which maintained ARF6 activity in an APE1-redox-dependent manner, promoting its ability to regulate MMP-14 recycling to the cell surface. In summary, these findings identify a novel redox-sensitive APE1-ARF6-MMP-14 signaling axis that mediates cellular invasion in esophageal carcinogenesis. SIGNIFICANCE: This study demonstrates the association between oxidative stress and the development and metastatic behavior of esophageal adenocarcinoma.


Adverse skin reactions to personal protective equipment during COVID-19 pandemic in Italian health care workers.
To avoid exposure to SARS-COV-2, healthcare professionals must use personal protective equipment (PPE). Their use has been related to a series of adverse effects; the most frequent adverse events were headache, dyspnoea, and pressure injuries. Skin adverse effects are very common, including contact dermatitis, itching, erythema, and acneiform eruptions. The objective of this study is to evaluate the skin problems caused by personal protection equipment (PPE) in health care workers (HCWs) and to individuate eventual risk factors. From May to June 2020 a retrospective observational multi-centric study conducted by an online survey sent by email, involving 10 hospital centers, was performed. We considered as independent variables gender and age, occupational group and sector, time of utilization, type and material of PPE. We tested 3 types of PPE: gloves, bonnet, and mask for different time of utilization (<1, 1-3, 3-6, >6 h). We performed a multiple logistic regression model to correlate them with skin adverse events occurrence. Among all the 1184 participants, 292 workers reported a dermatological pathology: 45 (15.41%) had psoriasis, 54 (18.49%) eczema, 38 (13.01%) acne, 48 (16.44%) seborrheic dermatitis, and 107 (36.64%) other. In our sample previous inflammatory dermatological conditions, female sex, prolonged use of PPE were significant risk factors for developing skin related adverse events considering all the PPE considered. The use of PPE is still mandatory in the hospital setting and skin adverse reactions still represent a global problem. Although data from Europe are limited, our study highlighted the importance of the problem of PPE skin reactions in a large sample of Italian healthcare professionals.


Predictors and consequences of global DNA methylation in cord blood and at three years.
DNA methylation changes have been implicated in many common chronic diseases leading to the hypothesis that environmental and age-related DNA methylation changes within individuals are involved in disease etiology. Few studies have examined DNA methylation changes within an individual over time and all of these studies have been conducted in adults. Here, we aim to characterize how global DNA methylation changes from birth to age three within a longitudinal birth cohort study and to determine whether there are consistent predictors of DNA methylation levels measured three years apart. We measured global DNA methylation in the same children at birth (cord blood) and again at three years of age among 165 children, using an immunoassay. We found that on average, DNA methylation was significantly higher in blood at age 3-years than in cord blood (p<0.01). However, for any individual child, the difference was less than would be expected by chance. We found that pre-pregnancy BMI was negatively predictive of both cord and three-year DNA methylation, even after statistical adjustment to account for the correlation between cord blood and three-year DNA methylation. The biologic implications of small changes in global DNA methylation are unknown. However, the observation that global DNA methylation levels persist within an individual from birth to age three supports the belief that factors that influence global DNA methylation, including pre-pregnancy BMI, may confer long-term effects.


Simultaneous characterization of glyco- and phosphoproteomes of mouse brain membrane proteome with electrostatic repulsion hydrophilic interaction chromatography.
Characterization of glyco- and phosphoproteins as well as their modification sites poses many challenges, the greatest being loss of their signals during mass spectrometric detection due to substoichiometric amounts and the ion suppression effect caused by peptides of high abundance. We report here an optimized protocol using electrostatic repulsion hydrophilic interaction chromatography for the simultaneous enrichment of glyco- and phosphopeptides from mouse brain membrane protein digest. With this protocol, we successfully identified 544 unique glycoproteins and 922 glycosylation sites, which were significantly higher than those from the commonly used hydrazide chemistry method (192 glycoproteins and 345 glycosylation sites). Moreover, a total of 383 phosphoproteins and 915 phosphorylation sites were recovered from the sample, suggesting that this protocol has the potential to enrich both glycopeptides and phosphopeptides simultaneously. Of the total 995 glycosylation sites identified from both methods, 96% were considered new as they were either annotated as putative or not documented in the newly released Swiss-Prot database. Thus, this study could be of significant value in complementing the current glycoprotein database and provides a unique opportunity to study the complex interaction of two different post-translational modifications in health and disease without being affected by interexperimental variations.


Investigation of relative micromotion at the stem-cement interface in total hip replacement.
Cemented total hip replacement has become a standard surgical technique to treat patients with osteoarthritis and osteonecrosis. The stem-cement interface experiences fretting wear in vivo due to low-amplitude oscillatory micromotion under physiological loading, and this wear is currently becoming important as a potential mechanism for the overall wear of cemented total hip replacements. However, the relative micromotion at the stem-cement interface has not been widely reported. In the present study, a new micromotion sensor is developed that is based on the deformation of a strain gauge, and this sensor is used to probe the migration of a polished Exeter stem within a Simplex P cement mantle through an in vitro wear simulation. It is demonstrated that the stem migration value generally increases with an increase in the number of loading cycles, with a gradual decrease of migration rate. Additionally, fretting wear is successfully replicated on the stem surface, and the micropores in the cement surface are considered to contribute to initiation and propagation of the fretting damage on the stem. This is confirmed by the observation that no evidence of fretting wear is detected on the stem where the surface is in contact with the pore-free areas on the cement. This study allows a deep insight into the micromotion at the stem-cement interface, and provides evidence highlighting the significance of the micropores in the cement surface in the generation of fretting wear on a polished femoral stem.


Type I, but not type II, interferon is critical in liver injury induced after ischemia and reperfusion.
We have documented the key role of toll-like receptor 4 (TLR4) activation and its signaling pathway mediated by interferon (IFN) regulatory factor 3, in the induction of inflammation leading to the hepatocellular damage during liver ischemia/reperfusion injury (IRI). Because type I IFN is the major downstream activation product of that pathway, we studied its role in comparison with IFN-gamma. Groups of type I (IFNAR), type II (IFNGR) IFN receptor-deficient mice, along with wild-type (WT) controls were subjected to partial liver warm ischemia (90 minutes) followed by reperfusion (1-6 hours). Interestingly, IFNAR knockout (KO) but not IFNGR KO mice were protected from IR-induced liver damage, as evidenced by decreased serum alanine aminotransferase and preservation of tissue architecture. IR-triggered intrahepatic pro-inflammatory response, assessed by tumor necrosis factor (TNF-alpha), interleukin 6 (IL-6), and chemokine (C-X-C motif) ligand 10 (CXCL-10) expression, was diminished selectively in IFNAR KO mice. Consistent with these findings, our in vitro cell culture studies have shown that: (1) although hepatocytes alone failed to respond to lipopolysaccharide (LPS), when co-cultured with macrophages they did respond to LPS via macrophage-derived IFN-beta; (2) macrophages required type I IFN to sustain CXCL10 production in response to LPS. This study documents that type I, but not type II, IFN pathway is required for IR-triggered liver inflammation/damage. Type I IFN mediates potential synergy between nonparenchyma and parenchyma cells in response to TLR4 activation.


Controlled multicenter evaluation of a bacteriophage-based method for rapid detection of Staphylococcus aureus in positive blood cultures.
Staphylococci are a frequent cause of bloodstream infections (BSIs). Appropriate antibiotic treatment for BSIs may be delayed because conventional laboratory testing methods take 48 to 72 h to identify and characterize isolates from positive blood cultures. We evaluated a novel assay based on bacteriophage amplification that identifies Staphylococcus aureus and differentiates between methicillin-susceptible and methicillin-resistant S. aureus (MSSA and MRSA, respectively) in samples taken directly from signal-positive Bactec blood culture bottles within 24 h of positive signal, with results available within 5 h. The performance of the MicroPhage KeyPath MRSA/MSSA blood culture test was compared to conventional identification and susceptibility testing methods. At four sites, we collectively tested a total of 1,165 specimens, of which 1,116 were included in our analysis. Compared to standard methods, the KeyPath MRSA/MSSA blood culture test demonstrated a sensitivity, specificity, positive predictive value, and negative predictive value of 91.8%, 98.3%, 96.3%, and 96.1%, respectively, for correctly identifying S. aureus. Of those correctly identified as S. aureus (n = 334), 99.1% were correctly categorized as either MSSA or MRSA. Analysis of a subset of the data revealed that the KeyPath MRSA/MSSA blood culture test delivered results a median of 30 h sooner than conventional methods (a median of 46.9 h versus a median of 16.9 h). Although the sensitivity of the test in detecting S. aureus-positive samples is not high, its accuracy in determining methicillin resistance and susceptibility among positives is very high. These characteristics may enable earlier implementation of appropriate antibiotic treatment for many S. aureus BSI patients.


Detailed investigations of dissolved hydrogen and hydrogen mass transfer in a biotrickling filter for upgrading biogas.
This study investigated the hydrogen mass transfer limitations in a biotrickling filter inoculated with hydrogenotrophic methanogens for biogas upgrading. A highly sensitive dissolved hydrogen probe allowed measuring concentrations in real-time. Experiments were conducted to test the mass transfer resistance in the gas and liquid films. Results demonstrated that the main resistance resides in the trickling liquid film and that promoting direct gas-biofilm mass transfer could improve upgrading performance by about 20%. Increasing the gas velocity (keeping a constant gas contact time) lowered the upgrading capacity. This was explained by the lowering of the concentration to the average concentration throughout the bed, which resulted a lower reaction rate. At extended gas contact times, the bioreactor shifted from microbial to diffusion limitation, causing lower upgrading capacities. Methane-containing biogas mimics (H2/CH4/CO2) were successfully upgraded to natural gas pipeline standards (>97% methane) with only minor performance reduction compared to upgrading just a H2/CO2 mixture.


Reduced speech perceptual acuity for stop consonants in individuals who stutter.
In individuals who stutter (IWS), speech fluency can be enhanced by altered auditory feedback, although it has adverse effects in control speakers. This indicates abnormalities in the auditory feedback loop in stuttering. Current motor control theories on stuttering propose an impaired processing of internal forward models that might be related to a blurred auditory-to-motor translation. Although speech sound perception is an essential skill to form internal models, perceptual acuity has not been studied in IWS so far. The authors tested the stability of phoneme percepts by analyzing participants' ability to identify voiced and voiceless stop consonants. Two syllable continua were generated by systematic modification of the voice onset time. The authors determined speech perceptual acuity by means of discriminatory power in 25 IWS and 24 matched control participants by determining the phoneme boundaries and by quantifying the interval of voice onset times for which phonemes were perceived ambiguously. In IWS, discriminatory performance was weaker and less stable over time when compared with control participants. In addition, phoneme boundaries were located at longer voice onset times in IWS. Persistent developmental stuttering is associated with less reliable phonological percepts, supporting current theories regarding the sensory-motor interaction in human speech.


Developmental regulation and neuronal expression of the mRNA of rat n-chimaerin, a p21rac GAP:cDNA sequence.
Human n-chimaerin is a GTPase-activating protein (GAP) for p21rac and a phorbol ester receptor. We have isolated rat n-chimaerin cDNA and investigated the cellular and developmental pattern of mRNA expression in the brain. There is extensive sequence conservation with human n-chimaerin in the coding region and the first 400 nucleotides of the 3'-untranslated region (UTR) (90% and 83% identity respectively). The rat cDNA encodes an additional 35 N-terminal amino acids compared with the reported human cDNA, which has a 5'-UTR sequence inversion and a 41-nucleotide deletion including the putative initiator methionine. The rat cDNA encodes a 334-amino acid protein (38200 M(r), pI 8.04) with 97% amino acid sequence identity with the human protein, after correction of the human 5'-DNA sequence. n-Chimaerin mRNA was detectable in embryonic rat brain at day 15 and increased in amount postnatally from birth to 20 days, coincident with cellular differentiation and synaptogenesis. n-Chimaerin mRNA is restricted to neurons, with highest concentrations in hippocampal pyramidal cells, granule cells of the dentate gyrus and cortical neurons. In the cerebellum the mRNA was detected only in Purkinje neurons. The pattern and specificity of mRNA expression suggests an important role for n-chimaerin in neuronal signal-transduction mechanisms.


Induction of lytic Epstein-Barr virus (EBV) infection in EBV-associated malignancies using adenovirus vectors in vitro and in vivo.
The consistent presence of EBV genomes in certain tumor types (in particular, AIDS-related central nervous system lymphomas and nasopharyngeal carcinomas) may allow novel, EBV-based targeting strategies. Tumors contain the latent (transforming) form of EBV infection. However, expression of either of the EBV immediate-early proteins, BZLF1 and BRLF1, is sufficient to induce lytic EBV infection, resulting in death of the host cell. We have constructed replication-deficient adenovirus vectors expressing the BZLF1 or BRLF1 immediate-early genes and examined their utility for killing latently infected lymphoma cells in vitro and in vivo. We show that both the BZLF1 and BRLF1 vectors efficiently induce lytic EBV infection in Jijoye cells (an EBV-positive Burkitt lymphoma cell line). Furthermore, lytic EBV infection converts the antiviral drug, ganciclovir (GCV), into a toxic (phosphorylated) form, which inhibits cellular as well as viral DNA polymerase. When Jijoye cells are infected with the BZLF1 or BRLF1 adenovirus vectors in the presence of GCV, viral reactivation is induced, but virus replication is inhibited (thus preventing the release of infectious EBV particles); yet cells are still efficiently killed. Finally, we demonstrate that the BZLF1 and BRLF1 adenovirus vectors induce lytic EBV infection when they are directly inoculated into Jijoye cell tumors grown in severe combined immunodeficiency mice. These results suggest that induction of lytic EBV infection in tumors, in combination with GCV, may be an effective strategy for treating EBV-associated malignancies.


Bentall procedure for the repair of a neoaortic aneurysm after the Norwood procedure in a patient with tricuspid atresia and a discordant ventriculo-arterial connection.
Neoaortic root dilation is a common phenomenon after the Norwood procedure, but the real incidence and its natural history are unclear. Regular surveillance in these patients after the operation is necessary. Herein, we present an 11-year-old boy born with tricuspid atresia, a discordant ventriculo-arterial connection and a hypoplastic aortic arch, who was palliated initially with a hybrid stage I procedure involving a reversed Blalock-Taussig shunt, followed by comprehensive stage II and then, an extra-cardiac fenestrated Fontan operation. The patient developed an aortic root aneurysm and severe aortic regurgitation. He was electively taken into the operating room, where cardiopulmonary bypass was established through a peripheral cannulation of the femoral vessels due to the high risk nature of the reoperation. A mechanical Bentall procedure was performed without residual lesions and the native ascending aorta was anastomosed as a single coronary button to the anterior wall of the graft.


[A variant of pathogenetic therapy in the acute period of perinatal hypoxic-traumatic brain lesions in premature infants].
The restorative potential of reparative cellular processes was studied in 39 immature infants born in hypoxia and receiving a pathogenetic treatment. In addition to the conventional treatment the test group of 24 infants received intravenously 20% piracetam solution, 1% riboflavin-mononucleotide solution, and 0.5% lipoic acid solution beginning from the first days of life. To correct the cellular metabolic processes in a control group of 15 infants, they were given 10% glucose solution, 5% ascorbic acid solution and cocarboxylase. The activity of dehydrogenases in the peripheral blood remained virtually unaltered in the control group but increased by an average of 1.5-2 times in the test group. An 18-month follow-up ascertained that the incidence of neurological complications in the infants receiving the proposed treatment was significantly lower as against the control patients.


Geo-epidemiology and autoimmunity.
Autoimmune disease (AD) affects approximately 3% of the population. This is an enormous number, but ironically the study of autoimmunity has not taken on the significance of many other diseases because so many of the ADs are relatively uncommon. Indeed, despite enormous advances in the diagnosis and the treatment of AD, there is still a paucity of data on the etiological events that lead to the clinical pathology. For most other human diseases, the etiology is addressed and often solved by the use of epidemiology. Epidemiology consists of the study of prevalence of a disease, coupled with analysis of genetic factors and detection of environmental agents. In the context of autoimmune conditions, preclinical epidemiology has recently been favored, as a consequence of the discovery that autoantibody precedes overt disease. The idea of a North-South gradient in the prevalence of ADs, with a reciprocal gradient in that of infectious injuries has proven to be debatable. More importantly, environmentally-induced changes have been shown to modify certain diseases giving rise to the key concept of epigenetics. However, it is clear that major voids exist. Some of these voids were hoped to be solved by the use of genome-wide associations. This, however, has proven very problematic, as the genetic basis of AD is considerably more complicated than once believed. We now base our hopes on next generational sequencing as a brut force undertaking to partially decipher the genetic code that predisposes individuals to AD. This volume is a compilation of papers in Autoimmunity Reviews and the Journal of Autoimmunity and presented as part of the 7th International Congress on Autoimmunity in Ljubljana, Slovenia. It is clearly impossible to present data on the geoepidemiology of all of the AD. Instead, we attempted to generate interest amongst immunologists to generate papers that are thought provoking but also contemporary reviews.


SNP and mRNA expression for glutathione peroxidase 4 in Kashin-Beck disease.
Kashin-Beck disease (KBD) is a chronic endemic osteoarthropathy, which mainly occurs in West and Northeast China. Epidemiological studies suggest that Se deficiency is an important environmental factor for the incidence of KBD. Glutathione peroxidase 4 (GPx4) belongs to the glutathione peroxidase family, which is crucial for optimal antioxidant defences. Our purpose is to investigate the putative association between GPx4 polymorphisms and the risk of KBD. Restriction fragment length polymorphism-PCR was used to detect two SNP (rs713041, rs4807542) in 219 cases and 194 controls in Han Chinese subjects, and quantitative analysis for the GPx4 mRNA level was performed by the real-time PCR method. The results revealed that linkage disequilibrium existed in the two SNP. A significant difference was observed in the haplotype A-T (P = 0·0066) of GPx4, which was obviously lower in the KBD cases (0·006 v. 0·032 %). Correlation analysis based on a single locus showed no association between each SNP and KBD risk. Furthermore, the GPx4 mRNA level was dramatically lower in the blood of KBD patients. Overall, our finding indicated GPx4 polymorphisms and decreased mRNA level may be related to the development of KBD in the Chinese population, suggesting GPx4 as a possible candidate susceptibility gene for KBD.


Markers of A1 astrocytes stratify to molecular sub-types in sporadic Creutzfeldt-Jakob disease brain.
Astrocytes are glial cells of the central nervous system that become reactive under conditions of stress. The functional properties of reactive astrocytes depend on their stimulus that induces the upregulation of specific genes. Reactive astrocytes are a neuropathological feature of prion disorders; however, their role in the disease pathogenesis is not well understood. Here, we describe our studies of one polarization state of reactive astrocytes, termed A1 astrocytes, in the frontal cortex region of 35 human sporadic Creutzfeldt-Jakob disease brains encompassing a range of molecular sub-types. Examination of two mRNA markers of A1 astrocytes, C3 and GBP2, revealed a strong linear correlation between the two following their log-normalization (P = 0.0011). Both markers were found upregulated in the sporadic Creutzfeldt-Jakob disease brain compared with age-matched control tissues (P = 0.0029 and 0.0002, for C3log and GBP2log, respectively), and stratifying samples based on codon 129 genotype revealed that C3log is highest in homozygous methionine and lowest in homozygous valine patients, which followed a linear trend (P = 0.027). Upon assessing other disease parameters, a significant positive correlation was found between GBP2log and disease duration (P = 0.031). These findings provide evidence for a divergence in the astrocytic environment amongst patients with sporadic Creutzfeldt-Jakob disease based on molecular sub-type parameters of disease. While more research will be needed to determine the global changes in the genomic profiles and resulting functional properties of reactive astrocytes in disease, considering the evidence demonstrating that A1 astrocytes harbour neurotoxic properties, the changes seen in C3log and GBP2log in the current study may reflect differences in pathogenic mechanisms amongst the sporadic Creutzfeldt-Jakob disease sub-types associated with the A1 polarization state.


An attempt to improve the therapeutic effect of boron neutron capture therapy using commonly employed 10B-carriers based on analytical studies on the correlation among quiescent tumor cell characteristics, tumor heterogeneity and cancer stemness.
Based on our previously published reports concerning the response of quiescent (Q) tumor cell populations to boron neutron capture therapy (BNCT), the heterogeneous microdistribution of 10B in tumors, which is influenced by the tumor microenvironment and the characteristics of the 10B delivery carriers, has been shown to limit the therapeutic effect of BNCT on local tumors. It was also clarified that the characteristics of 10B-carriers for BNCT and the type of combined treatment in BNCT can also affect the potential for distant lung metastases from treated local tumors. We reviewed the findings concerning the response of Q tumor cell populations to BNCT, mainly focusing on reports we have published so far, and we identified the mode of BNCT that currently offers the best therapeutic gain from the viewpoint of both controlling local tumor and suppressing the potential for distant lung metastasis. In addition, based on the finding that oxygenated Q tumor cells showed a large capacity to recover from DNA damage after cancer therapy, the interrelationship among the characteristics in Q tumor cell populations, tumor heterogeneity and cancer stemness was also discussed.


Surfing the internet with a BCI mouse.
In this paper, we present a new web browser based on a two-dimensional (2D) brain-computer interface (BCI) mouse, where our major concern is the selection of an intended target in a multi-target web page. A real-world web page may contain tens or even hundreds of targets, including hyperlinks, input elements, buttons, etc. In this case, a target filter designed in our system can be used to exclude most of those targets of no interest. Specifically, the user filters the targets of no interest out by inputting keywords with a P300-based speller, while keeps those containing the keywords. Such filtering largely facilitates the target selection task based on our BCI mouse. When there are only several targets in a web page (either an original sparse page or a target-filtered page), the user moves the mouse toward the target of interest using his/her electroencephalographic signal. The horizontal movement and vertical movement are controlled by motor imagery and P300 potential, respectively. If the mouse encounters a target of no interest, the user rejects it and continues to move the mouse. Otherwise the user selects the target and activates it. With the collaboration of the target filtering and a series of mouse movements and target selections/rejections, the user can select an intended target in a web page. Based on our browser system, common navigation functions, including history rolling forward and backward, hyperlink selection, page scrolling, text input, etc, are available. The system has been tested on seven subjects. Experimental results not only validated the efficacy of the proposed method, but also showed that free internet surfing with a BCI mouse is feasible.


First Report of Anthracnose on Pepper Fruit Caused by Colletotrichum scovillei in Brazil.
Anthracnose is major disease of pepper (Capsicum annum) in the tropics and causes severe damage both in the field and postharvest. In Brazil, this disease is caused by Colletotrichum acutatum, C. boninense, C. capsici, C. coccodes, and C. gloeosporioides, where the first species is responsible for 70% of all occurrences (3). Recently, C. acutatum has been considered a species complex (1); thus, the aim of this study was to verify the etiology of anthracnose on peppers using a morphological and molecular approaches. In 2011, pepper fruits with typical symptoms of anthracnose (dark, sunken spots with concentric rings of orange conidial masses) were collected in Viçosa, Minas Gerais, Brazil. A single spore isolate was obtained on potato dextrose agar (PDA), and the derived culture was deposited in the Coleção de Culturas de Fungos Fitopatogênicos "Prof. Maria Menezes" (code CMM-4200). The upper side colonies on PDA were gray, cotton-like, and pale gray to pale orange. Conidia were hyaline, aseptate, smooth, straight, cylindrical with round ends or occasionally with end ± acute, 12.5 to 17 μm long and 3.5 to 4 μm wide on synthetic nutrient deficient agar. The isolate was morphologically typical of species belonging to the C. acutatum complex. Molecular identification of the pathogen was carried out and sequences of the regions internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin (βt) were obtained and deposited in GenBank (Accession Nos. KJ541821 to KJ541823). A search in the Q-bank fungi database using the ITS, βt, and GAPDH sequences retrieved C. scovillei with 100% identity for all three genes. This pathogen was previously reported in Capsicum spp. only in Thailand, Indonesia, and Japan (1,2). To confirm pathogenicity, drops with 105 spores/ml were deposited in 10 artificially wounded fruits (cv. Itapuã 501 and Melina). In control fruits, drops of sterilized water were deposited onto wounds. The fruits were covered for one day with a transparent plastic bag with moisture supplied by a wet filter paper. The fruits were detached and mature. The bags were removed, and the fruits were incubated for 10 days in a growth chamber at 25°C with a photoperiod of 12 h. After 4 days, gray-brown to black sunken spots with concentric rings were observed on 100% of the wounded fruits that had been inoculated. No disease was observed on the control fruits. The fungus C. scovillei was successfully re-isolated from symptomatic fruits to fulfill Koch's postulates. To our knowledge, this is the first report of anthracnose on pepper fruit caused by C. scovillei in Brazil. Due to the diversity of species that cause anthracnose in Capsicum, future studies using morphological and molecular tools are essential for the correct identification of Colletotrichum spp. on pepper in Brazil. References: (1) U. Damm et al. Stud. Mycol. 73:37, 2012. (2) T. Kanto et al. J. Gen. Plant. Pathol. 80:73, 2014. (3) M. J. Z. Pereira et al. Hortic. Bras. 29:569, 2011.


Loss of the Bardet-Biedl protein Bbs1 alters photoreceptor outer segment protein and lipid composition.
Primary cilia are key sensory organelles whose dysfunction leads to ciliopathy disorders such as Bardet-Biedl syndrome (BBS). Retinal degeneration is common in ciliopathies, since the outer segments (OSs) of photoreceptors are highly specialized primary cilia. BBS1, encoded by the most commonly mutated BBS-associated gene, is part of the BBSome protein complex. Using a bbs1 zebrafish mutant, we show that retinal development and photoreceptor differentiation are unaffected by Bbs1-loss, supported by an initially unaffected transcriptome. Quantitative proteomics and lipidomics on samples enriched for isolated OSs show that Bbs1 is required for BBSome-complex stability and that Bbs1-loss leads to accumulation of membrane-associated proteins in OSs, with enrichment in proteins involved in lipid homeostasis. Disruption of the tightly regulated OS lipid composition with increased OS cholesterol content are paralleled by early functional visual deficits, which precede progressive OS morphological anomalies. Our findings identify a role for Bbs1/BBSome in OS lipid homeostasis, suggesting a pathomechanism underlying retinal degeneration in BBS.


A novel pleckstrin homology domain-containing protein enhances insulin-stimulated Akt phosphorylation and GLUT4 translocation in adipocytes.
Protein kinase B/Akt protein kinases control an array of diverse functions, including cell growth, survival, proliferation, and metabolism. We report here the identification of pleckstrin homology-like domain family B member 1 (PHLDB1) as an insulin-responsive protein that enhances Akt activation. PHLDB1 contains a pleckstrin homology domain, which we show binds phosphatidylinositol PI(3,4)P(2), PI(3,5)P(2), and PI(3,4,5)P(3), as well as a Forkhead-associated domain and coiled coil regions. PHLDB1 expression is increased during adipocyte differentiation, and it is abundant in many mouse tissues. Both endogenous and HA- or GFP-tagged PHLDB1 displayed a cytoplasmic disposition in unstimulated cultured adipocytes but translocated to the plasma membrane in response to insulin. Depletion of PHLDB1 by siRNA inhibited insulin stimulation of Akt phosphorylation but not tyrosine phosphorylation of IRS-1. RNAi-based silencing of PHLDB1 in cultured adipocytes also attenuated insulin-stimulated deoxyglucose transport and Myc-GLUT4-EGFP translocation to the plasma membrane, whereas knockdown of the PHLDB1 isoform PHLDB2 failed to attenuate insulin-stimulated deoxyglucose transport. Furthermore, adenovirus-mediated expression of PHLDB1 in adipocytes enhanced insulin-stimulated Akt and p70 S6 kinase phosphorylation, as well as GLUT4 translocation. These results indicate that PHLDB1 is a novel modulator of Akt protein kinase activation by insulin.


Selection and characterization of brivudin resistant bovine herpesvirus type 5.
Bovine herpesvirus type 5 (BoHV-5) is the agent of meningoencephalitis, an important disease of cattle in South America. The neuropathogenesis of BoHV-5 infection is poorly understood and most previous research focused on the role of envelope glicoproteins in neurovirulence. Thymidine kinase (TK) is a viral enzyme necessary for virus replication in neurons and, therefore, represents a potential target for virus attenuation. The selection and characterization of BoHV-5 variants resistant to the nucleoside analog brivudin (BVDU), which selects TK-defective viruses is here described. Several BVDU-resistant clones were obtained after multiple passages in tissue culture in the presence of BVDU and one clone (BoHV-5/R-27) was further characterized. The selected clone replicated to similar titers and produced plaques with similar size and morphology to those of wild-type virus (SV507/99). The genetic stability of the resistant virus was demonstrated after ten passages in cell culture in the absence of the drug. Moreover, the drug-resistant virus showed reduced virulence in a rabbit model: virus inoculation in four rabbits did not result in disease, in contrast with 75% morbidity (3/4) and 50% mortality (2/2) among rabbits inoculated with the parental virus. These results demonstrate that BoHV-5 is sensitive to BVDU and that drug-resistant mutants can be readily selected upon BVDU treatment. BVDU-resistant mutants, likely defective in TK, retained their ability to replicate in tissue culture yet were attenuated for rabbits. This strategy to obtain TK-defective BoHV-5 may be useful to study the role of TK in BoHV-5 neuropathogenesis and for vaccine development.


The anticipatory proportion as an indicator of language impairment in early-stage cognitive disorder in the elderly.
The anticipatory proportion (AP), the ratio between perseverative and anticipatory speech errors, is reduced in patients with brain injury. However, it is unknown whether the AP is also reduced in elderly speakers with cognitive impairment. 20 elderly speakers with a Mini Mental State Examination (MMSE) score of 25-27 and 20 elderly speakers with an MMSE score of 28-30 were assessed using a tongue-twister-based speech test, the Regensburg Word Fluency Test (RWT) and an object naming test. The AP in the group of speakers with an MMSE score of 25-27 was significantly lower. Accordingly, the AP and scores in the RWT and the object naming test were higher in persons with an MMSE score of 28-30. Language alterations in mild cognitive dysfunction are detectable with the AP. Further longitudinal studies are needed to evaluate the predictive value of the AP.


Clinical profile and impact on quality of life: seven years experience with patients of alopecia areata.
Alopecia areata (AA) is the most common cause of localized, non-scarring alopecia. Stress and other psychological factors have been implicated in the causation of the disease, and it is also found to alter the course of the disease process. Unfortunately no one has studied the impact of AA on the quality of life, which includes the social life of the patients. To study the clinical profile and impact of alopecia areata on the quality of life, including the social life of adult patients with severe forms of the disease. The present study determined the clinical pattern of AA and its impact on the quality of life (QOL) in all the patients with severe forms of alopecia areata attending the Dermatology Outpatient Department. The male : female ratio was 1.86 : 1. Most (58.03%) of the patients were between 21 and 40 years of age. Almost 40% of the patients had associated systemic disease or other dermatological disorders. A family history of AA was found in 593 (20.02%) of the patients. Nail changes were observed in 297 (10%) of the patients. There were significant differences between the mean Dermatology Life Quality Index (DLQI) score in cases with severe forms of AA and controls ( P < 0.001). Severe forms of alopecia areata had a major impact on the psychosocial well-being of the patients. These individuals had to be treated early, and they required more than just prescription drugs. Educational and psychological support in addition to medical therapy for AA could improve their long-term physical outcomes.