Robot-assisted retroperitoneal lymph node dissection: Initial experience in Japan.
For patients with testicular tumors who need the surgical management, open retroperitoneal lymph node dissection (O-RPLND) is considered the gold standard treatment. However, recently, robot-assisted RPLND (R-RPLND) has gained popularity as a minimally invasive therapy alternative to O-RPLND and laparoscopic RPLND. Here, we report the case of a 32-year-old man presenting with a left testicular teratoma with several enlarged left para-aortic lymph nodes. After the orchiectomy, the patient underwent R-RPLND with an operation time of 279 min, console time of 189 min, bleeding volume of 59 mL, and no significant complications, resulting in a successful outcome. To the best of our knowledge, this is the first reported case of R-RPLND in Japan. Based on our experience, R-RPLND may provide safe and effective outcomes; however, further research is required before the widespread implementation of this technique.


Removing label ambiguity in learning-based visual saliency estimation.
Visual saliency is a useful clue to depict visually important image/video contents in many multimedia applications. In visual saliency estimation, a feasible solution is to learn a "feature-saliency" mapping model from the user data obtained by manually labeling activities or eye-tracking devices. However, label ambiguities may also arise due to the inaccurate and inadequate user data. To process the noisy training data, we propose a multi-instance learning to rank approach for visual saliency estimation. In our approach, the correlations between various image patches are incorporated into an ordinal regression framework. By iteratively refining a ranking model and relabeling the image patches with respect to their mutual correlations, the label ambiguities can be effectively removed from the training data. Consequently, visual saliency can be effectively estimated by the ranking model, which can pop out real targets and suppress real distractors. Extensive experiments on two public image data sets show that our approach outperforms 11 state-of-the-art methods remarkably in visual saliency estimation.


Epidermal-growth-factor receptor correlates negatively with cell density in cervical squamous epithelium and is down-regulated in cancers of the human uterus.
The role of the epidermal-growth-factor receptor (EGFR) in cervical cancer is controversial, due to technical difficulties in localizing or in quantifying EGFR by homogenate assays or immunohistochemistry. Our autoradiographic approach, in combination with morphometry, allowed cell-type-specific quantification of EGFR, leading to the following observations: (i) In normal cervical epithelium, EGFR levels per cell were high in non-dividing squamous cells of the upper layers of normal epithelium, where a mitogenic function of these EGFRs can be excluded. (ii) In contrast to earlier findings in tissue homogenates, but consistent with our observation in normal cervical epithelium that cells of the proliferating strata (basal and parabasal cells) express intermediate and comparatively reduced levels of EGFR per cell, cervical cancers displayed a significant reduction both of specific EGF binding and of EGFR levels per cell as compared with normal epithelium. (iii) A significant negative correlation of cell density and EGFR number per cell was obtained. In normal cervical epithelium, cervical intra-epithelial neoplasia and invasive cervical cancer (p = 0.002). This negative correlation was most evident in normal epithelium, where large changes of cell density occur within one slide (p < 0.001). (iv) Specific EGF-binding was also significantly reduced in endometrial cancers when compared with normal endometrium. It is proposed that in uterine tissues low or intermediate levels of EGFR do not exclude their function as mediators of cell proliferation.


Short dual antiplatelet therapy followed by P2Y12 inhibitor monotherapy vs. prolonged dual antiplatelet therapy after percutaneous coronary intervention with second-generation drug-eluting stents: a systematic review and meta-analysis of randomized clinical trials.
After percutaneous coronary intervention (PCI) with second-generation drug-eluting stent (DES), whether short dual antiplatelet therapy (DAPT) followed by single antiplatelet therapy (SAPT) with a P2Y12 receptor inhibitor confers benefits compared with prolonged DAPT is unclear. Multiple electronic databases, including PubMed, Scopus, Web of Sciences, Ovid, and ScienceDirect, were searched to identify randomized clinical trials comparing ≤3 months of DAPT followed by P2Y12 inhibitor SAPT vs. 12 months of DAPT after PCI with second-generation DES implantation. The primary and co-primary outcomes of interest were major bleeding and stent thrombosis 1 year after randomization. Summary hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated by fixed-effect and random-effects models. Multiple sensitivity analyses including random-effects models 95% CI adjustment were applied. A sensitivity analysis comparing trials using P2Y12 inhibitor SAPT with those using aspirin SAPT was performed. A total of five randomized clinical trials (32 145 patients) were available. Major bleeding was significantly lower in the patients assigned to short DAPT followed by P2Y12 inhibitor SAPT compared with those assigned to 12-month DAPT (random-effects model: HR 0.63, 95% 0.45-0.86). No significant differences between groups were observed in terms of stent thrombosis (random-effects model: HR 1.19, 95% CI 0.86-1.65) and the secondary endpoints of all-cause death (random-effects model: HR 0.85, 95% CI 0.70-1.03), myocardial infarction (random-effects model: HR 1.05, 95% CI 0.89-1.23), and stroke (random-effects model: HR 1.08, 95% CI 0.68-1.74). Sensitivity analyses showed overall consistent results. By comparing trials testing ≤3 months of DAPT followed by P2Y12 inhibitor SAPT vs. 12 months of DAPT with trials testing ≤3 months of DAPT followed by aspirin SAPT vs. 12-month of DAPT, there was no treatment-by-subgroup interaction for each endpoint. By combining all these trials, regardless of the type of SAPT, short DAPT was associated with lower major bleeding (random-effects model: HR 0.63, 95% CI 0.48-0.83) and no differences in stent thrombosis, all-cause death, myocardial infarction, and stroke were observed between regimens. After second-generation DES implantation, 1-3 months of DAPT followed by P2Y12 inhibitor SAPT is associated with lower major bleeding and similar stent thrombosis, all-cause death, myocardial infarction, and stroke compared with prolonged DAPT. Whether P2Y12 inhibitor SAPT is preferable to aspirin SAPT needs further investigation.


[Significance of the expression of nucleolar argyrophilic proteins and antigen Ki-67 in the evaluation of cell proliferative activity and in the prediction of minimal (T1) lung cancer].
S u m m a ry. - The subject of the study was 20 cases of non-small-cell lung carcinomas, up to 3 cm in diameter, conventionally designed as minimal lung cancers removed in patients operated on at the N. N. Blokhin Cancer Research Centre, Russian Academy of Medical Sciences in 1986 to 2001. According to survival rates after surgery, the patients were divided into two groups: 1) those who died within the first two years; 2) those who were followed up for 3-5 years. Histological, histochemical, and immunohistochemical studies were performed. The expression of argyrophylic nucleolar organizer site proteins (Ag-NOS-proteins) that characterized the rate of cell proliferation (the duration of a cellular cycle) and the expression of Ki-67 antigen, which reflected the fraction of growth (the number of proliferating cells), were revealed in the tumor cells. Minimal lung cancers were found to be a heterogeneous group of neoplasms showing differences in both the rate of cell proliferation and the count of proliferating cells. The cell proliferation rate is a determinant of the clinical course of minimal lung cancers. Group 1 tumors characterized by the superexpression of Ag-NOS-proteins and, accordingly, the higher cell proliferation rate and the moderate count of proliferating cells had a poor prognosis even in the presence of Stage IA whereas Group 2 tumors with a large quantity of proliferating cells, but with the less rate of cell proliferation were characterized by a much better prognosis. The rate of cell proliferation (expression of Ag-NOS-proteins) and the count of proliferating cells (the expression of Ki-67 antigen) should be simultaneously studied to have more complete information on the proliferative potential of tumor cells and on the prediction of the course of neoplasms.


Gut microbiota and cancer immunotherapy: prognostic and therapeutic implications.
The immune checkpoint inhibitors have opened new horizons in oncology. Although the indications for the use of Immune checkpoint inhibitors in cancer patients are expanding, there is still a need for markers that can aid in patient selection. Gastrointestinal microbiota can be among these markers. Recently, gastrointestinal microbiota stated to have a bidirectional relation with cancer immunotherapy with roles in both prognostic and therapeutic sides. Preclinical data suggest that modulation of the microbiota could become a novel strategy for improving the efficacy of immunotherapy. However, its labile structure prone to be affected by many factors. Further research can delineate the mechanisms of the relationship between microbiota and immunotherapy can have clinical implications.


Outcome of bilateral periventricular nodular heterotopia in monozygotic twins with megalencephaly.
Bilateral periventricular nodular heterotopia (BPNH) is a neuronal migration disorder, characterized by grey-matter cellular rests in the periventricular regions. Patients frequently present with seizures during childhood or later in life. Monozygotic twin sisters were found to have enlarged head circumferences (>95th centile) during a routine medical visit at 6 months of age. A brain CT scan revealed grey-matter cellular rests in the subependymal regions extending to the subcortical white matter. In some areas these cell aggregates had nodular margins, consistent with the diagnosis of BPNH. At 6 years of age they are cognitively and neurologically normal. They have not had any seizures and their heads continue to grow along the 95th centile.


Radial secondary electron dose profiles and biological effects in light-ion beams based on analytical and Monte Carlo calculations using distorted wave cross sections.
To speed up dose calculation, an analytical pencil-beam method has been developed to calculate the mean radial dose distributions due to secondary electrons that are set in motion by light ions in water. For comparison, radial dose profiles calculated using a Monte Carlo technique have also been determined. An accurate comparison of the resulting radial dose profiles of the Bragg peak for (1)H(+), (4)He(2+) and (6)Li(3+) ions has been performed. The double differential cross sections for secondary electron production were calculated using the continuous distorted wave-eikonal initial state method (CDW-EIS). For the secondary electrons that are generated, the radial dose distribution for the analytical case is based on the generalized Gaussian pencil-beam method and the central axis depth-dose distributions are calculated using the Monte Carlo code PENELOPE. In the Monte Carlo case, the PENELOPE code was used to calculate the whole radial dose profile based on CDW data. The present pencil-beam and Monte Carlo calculations agree well at all radii. A radial dose profile that is shallower at small radii and steeper at large radii than the conventional 1/r(2) is clearly seen with both the Monte Carlo and pencil-beam methods. As expected, since the projectile velocities are the same, the dose profiles of Bragg-peak ions of 0.5 MeV (1)H(+), 2 MeV (4)He(2+) and 3 MeV (6)Li(3+) are almost the same, with about 30% more delta electrons in the sub keV range from (4)He(2+)and (6)Li(3+) compared to (1)H(+). A similar behavior is also seen for 1 MeV (1)H(+), 4 MeV (4)He(2+) and 6 MeV (6)Li(3+), all classically expected to have the same secondary electron cross sections. The results are promising and indicate a fast and accurate way of calculating the mean radial dose profile.


Rapid Phenotypic and Metabolomic Domestication of Wild Penicillium Molds on Cheese.
Fermented foods provide novel ecological opportunities for natural populations of microbes to evolve through successive recolonization of resource-rich substrates. Comparative genomic data have reconstructed the evolutionary histories of microbes adapted to food environments, but experimental studies directly demonstrating the process of domestication are lacking for most fermented food microbes. Here, we show that during adaptation to cheese, phenotypic and metabolomic traits of wild Penicillium molds rapidly change to produce domesticated phenotypes with properties similar to those of the industrial cultures used to make Camembert and other bloomy rind cheeses. Over a period of just a few weeks, populations of wild Penicillium strains serially passaged on cheese had reduced pigment, spore, and mycotoxin production. Domesticated strains also had a striking change in volatile metabolite production, shifting from production of earthy or musty volatile compounds (e.g., geosmin) to fatty and cheesy volatiles (e.g., 2-nonanone, 2-undecanone). RNA sequencing demonstrated a significant decrease in expression of 356 genes in domesticated strains, with an enrichment of many secondary metabolite production pathways in these downregulated genes. By manipulating the presence of neighboring microbial species and overall resource availability, we demonstrate that the limited competition and high nutrient availability of the cheese environment promote rapid trait evolution of Penicillium molds.IMPORTANCE Industrial cultures of filamentous fungi are used to add unique aesthetics and flavors to cheeses and other microbial foods. How these microbes adapted to live in food environments is generally unknown as most microbial domestication is unintentional. Our work demonstrates that wild molds closely related to the starter culture Penicillium camemberti can readily lose traits and quickly shift toward producing desirable aroma compounds. In addition to experimentally demonstrating a putative domestication pathway for P. camemberti, our work suggests that wild Penicillium isolates could be rapidly domesticated to produce new flavors and aesthetics in fermented foods.


Single Cell Portal: an interactive home for single-cell genomics data.
Single-cell omics research has the power to leave a deep impact on modern healthcare. Sharing data widely and freely advances this progress in both the academic and clinical spheres. We developed the Single Cell Portal (SCP) to maximize the impact of this work. SCP enables data sharing, supports dynamic results visualization, and facilitates scientific exploration across a large repository of single-cell datasets. SCP's data contributors maintain full control over how their data are shared and presented, without requiring web development expertise. Finally, SCP supports the entire lifecycle of a research project, from sparking an idea, to fine-tuning the data with collaborators, to sharing results in an accessible and interactive way. This paper highlights the most valuable ways in which SCP helps to advance single-cell research.


Vascular Cell Adhesion Molecule 1, Intercellular Adhesion Molecule 1, and Cluster of Differentiation 146 Levels in Patients with Type 2 Diabetes with Complications.
Type 2 diabetes mellitus (T2DM) is a multisystemic, chronic disease accompanied by microvascular complications involving various complicated mechanisms. Intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and cluster of differentiation-146 (CD146) are mainly expressed by endothelial cells, and facilitate the adhesion and transmigration of immune cells, leading to inflammation. In the present study, we evaluated the levels of soluble adhesion molecules in patients with microvascular complications of T2DM. Serum and whole blood samples were collected from 58 T2DM patients with microvascular complications and 20 age-matched healthy subjects. Levels of soluble ICAM-1 (sICAM-1) and soluble VCAM-1 (sVCAM-1) were assessed using enzyme-linked immunosorbent assay, while flow cytometry was used to determine CD146 levels. Serum sICAM-1 levels were lower in T2DM patients with microvascular complications than in healthy controls (P<0.05). No significant differences were found in sVCAM-1 and CD146 levels between the study and the control group. Although patients were subdivided into groups according to the type of microvascular complications that they experienced, cell adhesion molecule levels were not correlated with the complication type. In the study group, most of the patients were on insulin therapy (76%), and 95% of them were receiving angiotensin-converting enzyme (ACE)-inhibitor agents. Insulin and ACE-inhibitors have been shown to decrease soluble adhesion molecule levels via various mechanisms, so we suggest that the decreased or unchanged levels of soluble forms of cellular adhesion molecules in our study group may have resulted from insulin and ACE-inhibitor therapy, as well as tissue-localized inflammation in patients with T2DM.


Two types of muscarinic response to acetylcholine in mammalian cortical neurons.
Applications of acetylcholine (AcCho) to pyramidal cells of guinea pig cingulate cortical slices maintained in vitro result in a short latency inhibition, followed by a prolonged increase in excitability. Cholinergic inhibition is mediated through the rapid excitation of interneurons that utilize the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). This rapid excitation of interneurons is associated with a membrane depolarization and a decrease in neuronal input resistance. In contrast, AcCho-induced excitation of pyramidal cells is due to a direct action that produces a voltage-dependent increase in input resistance. In the experiments reported here, we investigated the possibility that these two responses are mediated by different subclasses of cholinergic receptors. The inhibitory and slow excitatory responses of pyramidal neurons were blocked by muscarinic but not by nicotinic antagonists. Pirenzepine was more effective in blocking the AcCho-induced slow depolarization than in blocking the hyperpolarization of pyramidal neurons. The two responses also varied in their sensitivity to various cholinergic agonists, making it possible to selectively activate either. These data suggest that AcCho may produce two physiologically and pharmacologically distinct muscarinic responses on neocortical neurons: slowly developing voltage-dependent depolarizations associated with an increase in input resistance in pyramidal cells and short-latency depolarizations associated with a decrease in input resistance in presumed GABAergic interneurons.


Investigating associations between physical activity-related neighborhood built environment features and child weight status to inform local practice.
Despite evidence of the importance of neighborhood built environment features in relation to physical activity and obesity, research has been limited in informing localized practice due to small sample sizes and limited geographic coverage. This demonstration study integrated data from a local pediatric health system with nationally available neighborhood built environment data to inform local decision making around neighborhood environments and childhood obesity. Height/weight from clinic visits and home neighborhood measures from the U.S. Environmental Protections Agency and WalkScore were obtained for 15,989 6-17 year olds. Multilevel models accounted for the nested data structure and were adjusted for neighborhood income and child sociodemographics. In 9-17 year olds, greater street connectivity and walkability were associated with a 0.01-0.04 lower BMIz (Ps = .009-.017) and greater residential density, street connectivity, and walkability were associated 5-7% lower odds of being overweight/obese (Ps = .004-.044) per standard deviation increase in environment variable. 45.9% of children in the lowest walkability tertile were overweight or obese, whereas 43.1% of children in the highest walkability tertile were overweight or obese. Maps revealed areas with low walkability and a high income-adjusted percent of children overweight/obese. In the Kansas City area, data showed that fewer children were overweight/obese in more walkable neighborhoods. Integrating electronic health records with neighborhood environment data is a replicable process that can inform local practice by highlighting the importance of neighborhood environment features locally and pointing to areas most in need of interventions.


Sustaining e-caring leadership in a post-pandemic world.
At the convergence of the "new normal" from the COVID-19 pandemic and the combination of caring, leadership, and technology, a new environment for organizational culture was formed. The injection of technology into how leaders provided care to their organization launched e-caring, and this is now considered a key in employee engagement and retention post COVID-19 pandemic. The purpose of this editorial is to briefly reflect on the importance of e-caring for leadership during the crisis and increase the commitment of nursing leaders to continue their great works even after the pandemic with an organizational culture infused with e-caring.


Community genomic analyses constrain the distribution of metabolic traits across the Chloroflexi phylum and indicate roles in sediment carbon cycling.
Sediments are massive reservoirs of carbon compounds and host a large fraction of microbial life. Microorganisms within terrestrial aquifer sediments control buried organic carbon turnover, degrade organic contaminants, and impact drinking water quality. Recent 16S rRNA gene profiling indicates that members of the bacterial phylum Chloroflexi are common in sediment. Only the role of the class Dehalococcoidia, which degrade halogenated solvents, is well understood. Genomic sampling is available for only six of the approximate 30 Chloroflexi classes, so little is known about the phylogenetic distribution of reductive dehalogenation or about the broader metabolic characteristics of Chloroflexi in sediment. We used metagenomics to directly evaluate the metabolic potential and diversity of Chloroflexi in aquifer sediments. We sampled genomic sequence from 86 Chloroflexi representing 15 distinct lineages, including members of eight classes previously characterized only by 16S rRNA sequences. Unlike in the Dehalococcoidia, genes for organohalide respiration are rare within the Chloroflexi genomes sampled here. Near-complete genomes were reconstructed for three Chloroflexi. One, a member of an unsequenced lineage in the Anaerolinea, is an aerobe with the potential for respiring diverse carbon compounds. The others represent two genomically unsampled classes sibling to the Dehalococcoidia, and are anaerobes likely involved in sugar and plant-derived-compound degradation to acetate. Both fix CO2 via the Wood-Ljungdahl pathway, a pathway not previously documented in Chloroflexi. The genomes each encode unique traits apparently acquired from Archaea, including mechanisms of motility and ATP synthesis. Chloroflexi in the aquifer sediments are abundant and highly diverse. Genomic analyses provide new evolutionary boundaries for obligate organohalide respiration. We expand the potential roles of Chloroflexi in sediment carbon cycling beyond organohalide respiration to include respiration of sugars, fermentation, CO2 fixation, and acetogenesis with ATP formation by substrate-level phosphorylation.


Effects of pesticides on the fatty acid and phospholipid composition of Escherichia coli.
Cells of Escherichia coli contained an altered phospholipid and fatty acid composition when grown in the presence of some pesticides. Whereas parathion increased the concentration of all phospholipid species without changes in their polar head groups. DDT (dichlorodiphenyltrichloroethane) decreased the proportion of neutral serine-derived phosphatides and dieldrin decreased the proportion of negatively charged phospholipids. The saturated/unsaturated plus cyclopropane fatty acid ratio was increased in all cases. The changes suggested that cells adapted their membrane lipids to compensate for the presence of pesticides in the environment.


Cortical networks of hemianopia stroke patients: a graph theoretical analysis of EEG signals at resting state.
Visual cortical stroke patients may have hemianopia symptom, which affects a number of visual functions. Most studies on hemianopia stroke have mainly focused on cortical activation during visual stimulation, leaving the pattern of functional connectivity between different brain regions uncovered yet. In the present study, we investigate the resting neural networks of hemianopia stroke patients by graph theoretical analysis of functional brain networks constructed with phase synchronization indexes of multichannel electroencephalography (EEG) signals. Our results showed that although the global network topological metrics, i.e., weighted clustering coefficient and characteristic path length of patients and healthy controls are comparable, the left primary visual cortex of patients tend to be less active than that of age-matched healthy subjects. However, hemianopia patients showed greater activation in the ipsilesional (left) temporopolar and orbit frontal areas and the contralesional (right) associative visual cortex. These results may offer new insight into neural substrates of the hemianopia stroke, and the further study of neural plasticity and brain reorganization after hemianopia.


Vascular density has prognostic value in Merkel cell carcinoma.
Merkel cell carcinomas are aggressive tumours for which histological prognostic factors need to be established. This study examines the prognostic role of vascular density, based on CD34 immunohistochemical staining in Merkel cell carcinoma. Thirty-six cases of Merkel cell carcinoma were immunohistochemically stained for the endothelial marker CD34. Vascular density was assessed in the tumor and stroma with a Chalkley eyepiece graticule. The scores of vascular density were correlated with other clinical and histological parameters to determine the prognostic significance of tumor vascularity. Increased vascular density was shown to be significantly associated with a worse prognosis (P = 0.005). A 1-unit increase in total vessel score was associated with a 3.9 times increase in the risk of death (95% hazard ratio confidence limits 1.50-10.32). Other factors associated with a worse outcome included tumor size (P = 0.05), the presence of lymphovascular invasion (P = 0.03), and tumor mast cell count (P < 0.002). Increased vascular density is associated with a worse prognosis in Merkel cell carcinomas. Assessment of vascular density may assist in predicting clinical behavior in these tumors and in evaluating the effects of adjuvant therapy.


Hospital revisits within 30 days after discharge for medical conditions targeted by the Hospital Readmissions Reduction Program in the United States: national retrospective analysis.
To determine any changes in total hospital revisits within 30 days of discharge after a hospital stay for medical conditions targeted by the Hospital Readmissions Reduction Program (HRRP). Retrospective cohort study. Hospital stays among Medicare patients for heart failure, acute myocardial infarction, or pneumonia between 1 January 2012 and 1 October 2015. Medicare fee-for-service patients aged 65 or over. Total hospital revisits within 30 days of discharge after hospital stays for medical conditions targeted by the HRRP, and by type of revisit: treat-and-discharge visit to an emergency department, observation stay (not leading to inpatient readmission), and inpatient readmission. Patient subgroups (age, sex, race) were also evaluated for each type of revisit. Our study cohort included 3 038 740 total index hospital stays from January 2012 to September 2015: 1 357 620 for heart failure, 634 795 for acute myocardial infarction, and 1 046 325 for pneumonia. Counting all revisits after discharge, the total number of hospital revisits per 100 patient discharges for target conditions increased across the study period (monthly increase 0.023 visits per 100 patient discharges (95% confidence interval 0.010 to 0.035)). This change was due to monthly increases in treat-and-discharge visits to an emergency department (0.023 (0.015 to 0.032) and observation stays (0.022 (0.020 to 0.025)), which were only partly offset by declines in readmissions (-0.023 (-0.035 to -0.012)). Increases in observation stay use were more pronounced among non-white patients than white patients. No significant change was seen in mortality within 30 days of discharge for target conditions (-0.0034 (-0.012 to 0.0054)). In the United States, total hospital revisits within 30 days of discharge for conditions targeted by the HRRP increased across the study period. This increase was due to a rise in post-discharge emergency department visits and observation stays, which exceeded the decline in readmissions. Although reductions in readmissions have been attributed to improvements in discharge planning and care transitions, our findings suggest that these declines could instead be because hospitals and clinicians have intensified efforts to treat patients who return to a hospital within 30 days of discharge in emergency departments and as observation stays.


Second line molecular diagnosis for bovine tuberculosis to improve diagnostic schemes.
Surveillance of bovine tuberculosis (bTB) is partly based on the sanitary inspection of carcasses at the abattoir to detect bTB-like lesions which, in compliance with EU recommendations, are analysed by bacteriology and histopathology to disclose Mycobacterium bovis (or M. caprae) infection. Moreover, since 2012, a PCR method with similar sensitivity and specificity values of histopathology and bacteriology respectively is additionally employed in France, partially compensating for the weaknesses of classical diagnostic methods. We analysed a collection of bTB-like lesions from cattle presenting positive histological results albeit with negative PCR results. We present here the results of these samples, recovered from 292 animals culled between 2013 and 2016, analysed with a second line molecular diagnosis approach that consists in a combination of PCRs targeting the M. tuberculosis-M. avium complexes as well as the Mycobacterium genus and sequencing of hsp65 gene. These molecular analyses disclosed to identify the presence of non-tuberculous bacteria which could be responsible for most of these non-specific TB lesions: non tuberculous mycobacteria (24%) or Actinomycetales (56%) such as Rhodococcus equi (53%); 24% of the samples were negative. M. bovis -or any other MTBC members- was neither detected by molecular methods nor isolated in any of them at the end of the 3 months of culture. In conclusion, these results highlight the lack of specificity of histopathology and the usefulness of a first line PCR with a second line molecular diagnostic test to circumvent it. This diagnostic strategy makes it possible to reduce the number of suspect bTB cases raised at the abattoir or shortening their lock-up periods. By simplifying diagnostic schemes, the use of this tool could improve bTB surveillance and make eradication programs more efficient in the future.


External Validity of the T-SAFT90: A Soccer Simulation Including Technical and Jumping Activities.
To examine the physiological, muscle-damage, endocrine, and immune responses to a modified soccer-simulation protocol to include technical and jumping activities characteristic of match play (the Technical Soccer-Specific Aerobic Field Test; T-SAFT90). Eighteen university players (age 23 [2] y, stature 175 [5] cm, body mass 74 [11] kg) performed the 90-minute protocol, with acute physiological responses monitored via heart rate, ratings of perceived exertion (6-20 scale), and body mass changes. Creatine kinase, myoglobin, cortisol, and leukocyte subset concentrations were measured at baseline, immediately (0 h), and 24 hours post T-SAFT90. T-SAFT90 incurred an average heart rate equivalent to 87% (5%) of maximum, 16 (2) a.u. ratings of perceived exertion, and a 1.5% (1.0%) body mass deficit. Moderate to large proliferation of leukocyte subsets (P ≤ .01; leukocytes: 6.4-fold; neutrophils: 5.5-fold; lymphocytes: 2.0-fold) and increases in cortisol (2.3-fold) were observed at 0 hours (effect size = 1.13-3.52), each returning to baseline by 24 hours (P > .45; effect size = 0.05-0.50). Myoglobin peaked immediately post T-SAFT90 (4.8-fold), whereas creatine kinase (24 h: 6.0-fold) showed a delayed time course (both P ≤ .001; very large effects; effect size = 2.66 and 3.43, respectively). The magnitude and time course of the physiological, immune, endocrine, and muscle-damage markers observed during and following T-SAFT90 are similar to values reported in match-play literature, demonstrating external validity of the simulation.


Meat proteins in a high-fat diet have a substantial impact on intestinal barriers through mucus layer and tight junction protein suppression in C57BL/6J mice.
Protein diets are well known for body maintenance and weight loss. However, it remains unclear whether and how different protein sources affect the intestinal epithelial integrity through tight junctions, mucus secretions and host immunity in diet-induced obesity. To evaluate possible effects, soybean, chicken and pork proteins either with low fat (12% kcal) or high fat (60% kcal) were administered to C57BL/6J mice for 12 weeks. Muc2 expression, tight junction proteins, goblet cells, and inflammatory cytokines in the colon and serum were measured. The intake of a high-fat pork protein diet decreased the number of goblet cells and inhibited Muc2 expression in the colon, which impaired the mucus barrier. Immunohistochemistry indicated decreased crypt depth and downregulation of tight junction proteins in high-fat diet fed mice, signifying losses of epithelial barriers. In addition, a pork protein diet reduces the key zonula occludens-1 and E-cadherin proteins. A high-fat meat protein diet induces colonic inflammatory injury by upregulating several key cytokines and increasing IL-1β, TNF-α, IL-6 and IFN-γ concentrations in serum. The intake of high-fat meat protein diets resulted in the impairment of the colon barrier through mucus suppression, downregulation of tight junctions, and gut inflammation in mice.


Time-domain estimation of the center frequency and spread of Doppler spectra in diagnostic ultrasound.
Two sets of estimators of the center frequency and mean square bandwidth of a Doppler spectrum are analyzed. The first set is based on a linear combination of the complex autocorrelation of the Doppler signal for different lags. It is shown that the argument (phase angle) of the complex autocorrelation function at unit sample lag is a close approximation to the mean Doppler shift, whereas its modulus (magnitude) gives information about the mean square bandwidth of the spectrum. Approximate expressions for the bias and variance of the estimators are derived, valid for long averaging times. The performances of the estimators are compared for signals with rectangular spectra and different signal-to-noise ratios.


Imaging features of primary and metastatic malignant perivascular epithelioid cell tumors.
The objective of our study was to describe the imaging features of primary and metastatic malignant perivascular epithelioid cell tumors (PEComas). In this retrospective study, 36 patients (26 women, 10 men; mean age, 53.1 years; age range, 35-77 years) with a histopathologically confirmed diagnosis of malignant PEComa who were seen at our institute between January 2007 and December 2012 were included. Pretreatment imaging of the primary tumor in 17 patients (CT, n = 13; MRI, n = 9; ultrasound, n = 5) and follow-up imaging in all 36 patients (CT, n = 36; MRI, n = 7) were reviewed by three radiologists in consensus. The most common site of presentation of malignant PEComas was the retroperitoneum (38.9%) followed by the female genital tract (27.8%). Kidneys (8/36) and uterus (8/36) were the organs most frequently involved. The mean largest dimension was 11.01 cm (range, 4.5-25 cm). Primary tumors were well-circumscribed masses (16/17), were heteroechoic on ultrasound (5/5), were hypodense (2/6) to isodense (4/6) enhanced significantly (11/12) and compared with the paraspinal muscles on CT, were hypointense to isointense compared with skeletal muscle on T1-weighted imaging (8/9), were heterogeneously hyperintense compared with skeletal muscle on T2-weighted imaging (9/9), and showed significant post-gadolinium enhancement (7/7) on MRI. Of the 17 primary tumors, necrosis was seen in seven tumors, hemorrhage in three tumors, and calcification in three tumors. Twenty-six of 36 (72%) patients developed metastases, most commonly to the lung (21.6%), liver (17.6%), and peritoneum (10.8%). Malignant PEComas in our study were large tumors that most often arise in the kidneys and uterus and metastasize to lungs, liver, or peritoneum. In our experience malignant PEComas should be considered in the differential diagnosis of large well-circumscribed renal and uterine tumors.


Complement-fixing antibody to Epstein-Barr virus soluble antigen in populations at high and low risk for nasopharyngeal carcinoma.
Greenland Eskimos comprise an ethnic group with one of the highest recorded incidence rates for nasopharyngeal carcinoma in the world. Sera from 625 Eskimos and 73 Danes (Caucasians) living in Greenland, as well as from 62 Danes living in Denmark, were tested for complement-fixing antibody to Epstein-Barr virus (EBV) soluble antigen and, from this study group, 129 donors were matched by age and sex for a study comparing antibody to viral capsid antigen, early antigen, and soluble antigen. Both Eskimos and Danes living in Greenland had significantly higher titers of EBV antibodies than Danes living in Denmark, suggesting that environment was more important than genetics or socio-economic factors in determining the antibody response to EBV. Age and sex were also factors, higher titers occurring in females and young Eskimos.


Impact of age on the facial expression of pain.
Old age has traditionally been viewed as being associated with a decline in emotional expressivity. Interestingly, empirical evidence based on analyses of facial expressions contradicts this traditionally view and points to absence of (or only very slight) age-related changes in emotional expressivity. However, this research on emotional expressivity in older persons has neglected one important emotionally colored state-expression of pain. In order to close this gap, we aimed to investigate the influence of age on the facial expression of pain. Forty young (mean age, 24.1 years) and 61 elderly (mean age, 72.3 years) subjects were investigated for their facial (Facial Action Coding System) and subjective responses to noxious mechanical and electrical stimuli of various intensities. Young and elderly subjects did not differ with respect to the frequency of facial responses during noxious mechanical and electrical stimulations. Moreover, age had no significant impact on the pain specificity of these facial responses. Furthermore, we found no significant age differences in self-report ratings of pressure and electrical pain, thus indicating that both age groups experienced comparable amounts of pain intensities. These findings suggest that the facial expression of pain, like facial expressions of other affective states, remains unchanged in older persons. Consequently, elderly individuals seem to communicate pain through their facial expression as validly as younger individuals do.


Temporal correlation of antibody responses to different epitopes of the human La autoantigen.
To investigate the temporal relationship of antibody responses to different La epitopes, sequential sera from nine patients with systemic lupus erythematosus and Sjogren's syndrome were tested by enzyme-linked immunosorbent assay for antibody binding to a series of recombinant fusion proteins containing different regions of the La molecule. The results of this analysis indicate that antibody responses to four different La fragments vary in parallel over time. This finding is supported by a statistical analysis indicating that the changes in antibody levels between the six pairs of responses were highly correlated (P less than 0.001). Furthermore, we show by immunoaffinity purification that antibodies to the three nonoverlapping La protein fragments do not cross-react with other fragments and, hence, represent independent populations. These results suggest that anti-La antibodies are coordinately produced to different epitopes on the La molecule, possibly reflecting an antigen-driven mechanism.


OXIDATIVE ASSIMILATION OF GLUCOSE BY PSEUDOMONAS AERUGINOSA.
Duncan, Margaret G. (The University of British Columbia, Vancouver, British Columbia, Canada) and J. J. R. Campbell. Oxidative assimilation of glucose by Pseudomonas aeruginosa. J. Bacteriol. 84:784-792. 1962-Oxidative assimilation of glucose by washed-cell suspensions of Pseudomonas aeruginosa was studied using C(14)-labeled substrate. At the time of glucose disappearance, only small amounts of radioactivity were present in the cells, and alpha-ketoglutaric acid accumulated in the supernatant fluid. Most of the material synthesized by the cells during oxidative assimilation was nitrogenous, the ammonia being supplied by the endogenous respiration. The cold trichloroacetic acid-soluble fraction and the lipid fraction appeared to be important during the early stages of oxidative assimilation, and the largest percentage of the incorporated radioactivity was found in the protein fraction. In the presence of added ammonia, assimilation was greatly increased and no alpha-ketoglutaric acid was found in the supernatant fluid. Sodium azide partially inhibited incorporation into all major cell fractions, and at higher concentrations depressed the rate of glucose oxidation. During oxidative assimilation, chloramphenicol specifically inhibited the synthesis of protein. Oxidative assimilation of glucose by this organism did not appear to involve the synthesis of a primary product such as is found in the majority of bacteria.


Positive affect and learning: exploring the "Eureka Effect" in dogs.
Animals may experience positive affective states in response to their own achievements. We investigated emotional responses to problem-solving in dogs, separating these from reactions to rewards per se using a yoked control design. We also questioned whether the intensity of reaction would vary with reward type. We examined the response (behavior and heart rate) of dogs as they learned to gain access to different rewards: (1) food (2) human contact, and (3) dog contact. Twelve beagles were assigned to matched pairs, and each dog served as both an experimental and a control animal during different stages of the experiment. We trained all dogs to perform distinct operant tasks and exposed them to additional devices to which they were not trained. Later, dogs were tested in a new context. When acting as an experimental dog, access to the reward was granted immediately upon completion of trained operant tasks. When acting as a control, access to the reward was independent of the dog's actions and was instead granted after a delay equal to their matched partner's latency to complete their task. Thus, differences between the two situations could be attributed to experimental dogs having the opportunity to learn to control access to the reward. Experimental dogs showed signs of excitement (e.g., increased tail wagging and activity) in response to their achievements, whereas controls showed signs of frustration (e.g., chewing of the operant device) in response to the unpredictability of the situation. The intensity of emotional response in experimental dogs was influenced by the reward type, i.e., greatest response to food and least to another dog. Our results suggest that dogs react emotionally to problem-solving opportunities and that tail wagging may be a useful indicator of positive affective states in dogs.


Pulmonary embolism after adult spinal deformity surgery.
Retrospective review. To determine the incidence and identify the associated risk factors of pulmonary embolism (PE) in patients who receive pharmacologic thromboprophylaxis after adult spinal deformity surgery. The risk of PE after adult spinal deformity surgery is reported to be as high as 2.2%. However, the incidence and associated risks of PE in the same patient population who receive postoperative pharmacologic thromboprophylaxis is unknown. The study included 361 adult patients with spinal deformity who underwent 407 corrective spinal procedures for scoliosis, kyphosis, or kyphoscoliosis. The incidence of PE was determined and compared with a study (historical control) of similar patients undergoing similar surgery but without postoperative pharmacologic thromboprophylaxis. Their demographic information, American Society of Anesthesiologists score, operative time, surgical approach, surgical complexity, and intraoperative blood loss were also analyzed to determine the presence of associated risk factors. Despite universal pharmacologic thromboprophylaxis, 10 pulmonary emboli (2.4%) were diagnosed. Patients undergoing anterior spinal surgery were at a significantly higher risk than those undergoing posterior spinal surgery (P = 0.024). The right-side anterior approach was also associated with a significantly higher incidence of PE compared with the left-sided anterior approach (P = 0.018). Although the rate of PE after posterior spinal surgery did not differ from the historical control, the rate of PE after anterior surgery was reduced by 50% compared with the historical control. Age, gender, estimated blood loss, operative time, revision status, and the number of fusion levels were not significant variables for PE. There were 2 epidural hematomas requiring decompression (0.48%) and 1 wound hematoma (0.24%). Although pharmacologic thromboprophylaxis probably does not have a role after posterior spinal surgery, the data in this study suggest that it does lower the incidence of PE after anterior spinal surgery.


Enhancing effects of contingency instructions on fear acquisition and extinction in anxiety disorders.
Explicit instructions regarding stimulus-threat associations increase acquisition and extinction of fear in healthy participants. The current study aimed to investigate the effect of contingency instructions on fear acquisition and extinction in patients with anxiety disorders. Patients with various anxiety disorders (N = 104) and healthy comparison participants (N = 93) participated in a differential fear conditioning task (within-subjects design). Approximately halfway through the acquisition phase, participants were instructed about the stimulus-threat association, and approximately halfway through the extinction phase, participants were informed that the unconditioned stimulus (US) would no longer be administered. Outcome measures were: fear-potentiated startle, skin conductance, fearfulness ratings, and US expectancy ratings. Patients demonstrated overall increased physiological and subjective fear responses during acquisition and extinction phases, relative to the comparison group. There were no major differences in fear acquisition and extinction between patients with different anxiety disorders. During acquisition, instructions led to increased discrimination of fear responses between a danger cue (conditioned stimulus [CS]+) and safety cue (CS-) in both patients and comparison participants. Moreover, instructions strengthened extinction of fear responses in the patient and comparison group. Patients and healthy comparison participants are better able to discriminate between danger and safety cues when they have been explicitly informed about cues that announce a threat situation. Considering the analogies between fear extinction procedures and exposure therapy, this suggests that specific instructions on stimulus-threat associations during exposure therapy might improve short-term treatment efficacy. The question remains for future studies whether instructions have a positive effect on extinction learning in the longer term. (PsycINFO Database Record


Rationale and design of a randomised controlled trial evaluating the effectiveness of an exercise program to improve the quality of life of patients with heart failure in primary care: The EFICAR study protocol.
Quality of life (QoL) decreases as heart failure worsens, which is one of the greatest worries of these patients. Physical exercise has been shown to be safe for people with heart failure. Previous studies have tested heterogeneous exercise programs using different QoL instruments and reported inconsistent effects on QoL. The aim of this study is to evaluate the effectiveness of a new exercise program for people with heart failure (EFICAR), additional to the recommended optimal treatment in primary care, to improve QoL, functional capacity and control of cardiovascular risk factors. Multicenter clinical trial in which 600 patients with heart failure in NYHA class II-IV will be randomized to two parallel groups: EFICAR and control. After being recruited, through the reference cardiology services, in six health centres from the Spanish Primary Care Prevention and Health Promotion Research Network (redIAPP), patients are followed for 1 year after the beginning of the intervention. Both groups receive the optimized treatment according to the European Society of Cardiology guidelines. In addition, the EFICAR group performs a 3 month supervised progressive exercise program with an aerobic (high-intensity intervals) and a strength component; and the programme continues linked with community resources for 9 months. The main outcome measure is the change in health-related QoL measured by the SF-36 and the Minnesota Living with Heart Failure Questionnaires at baseline, 3, 6 and 12 months. Secondary outcomes considered are changes in functional capacity measured by the 6-Minute Walking Test, cardiac structure (B-type natriuretic peptides), muscle strength and body composition. Both groups will be compared on an intention to treat basis, using multi-level longitudinal mixed models. Sex, age, social class, co-morbidity and cardiovascular risk factors will be considered as potential confounding and predictor variables. A key challenges of this study is to guarantee the safety of the patients; however, the current scientific evidence supports the notion of there being no increase in the risk of decompensation, cardiac events, hospitalizations and deaths associated with exercise, but rather the opposite. Safety assurance will be based on an optimized standardised pharmacological therapy and health education for all the participants. Clinical Trials.gov Identifier: NCT01033591.


Dif-Fusion: Toward High Color Fidelity in Infrared and Visible Image Fusion With Diffusion Models.
Color plays an important role in human visual perception, reflecting the spectrum of objects. However, the existing infrared and visible image fusion methods rarely explore how to handle multi-spectral/channel data directly and achieve high color fidelity. This paper addresses the above issue by proposing a novel method with diffusion models, termed as Dif-Fusion, to generate the distribution of the multi-channel input data, which increases the ability of multi-source information aggregation and the fidelity of colors. In specific, instead of converting multi-channel images into single-channel data in existing fusion methods, we create the multi-channel data distribution with a denoising network in a latent space with forward and reverse diffusion process. Then, we use the the denoising network to extract the multi-channel diffusion features with both visible and infrared information. Finally, we feed the multi-channel diffusion features to the multi-channel fusion module to directly generate the three-channel fused image. To retain the texture and intensity information, we propose multi-channel gradient loss and intensity loss. Along with the current evaluation metrics for measuring texture and intensity fidelity, we introduce Delta E as a new evaluation metric to quantify color fidelity. Extensive experiments indicate that our method is more effective than other state-of-the-art image fusion methods, especially in color fidelity. The source code is available at https://github.com/GeoVectorMatrix/Dif-Fusion.


Airborne microbial flora in a cattle feedlot.
A total of 1,408 cattle held in eight commercial feedlot pens were used to examine the quantity and diversity of microorganisms in cattle feedlot air. The effect of two feeding patterns on the generation of airborne dust and the total numbers of microorganisms was also examined (four feedlot pens/treatment). Microbial samples were collected, and dust particles that were 2.5 microm or less in diameter were measured with a Dustrak monitor during the evening dust peak for 4 days at sites both upwind and downwind of the feedlot pens. An Andersen biological cascade sampler was employed with different medium and incubation combinations for the capture and identification of bacteria and fungi. The results showed that when bacteria were considered, only nonpathogenic gram-positive organisms were recovered. However, gram-negative bacteria may have been present in a viable but nonculturable state. Fungi were recovered in smaller numbers than bacteria, and none of the fungi were pathogenic. The Dustrak results showed that one feeding pattern resulted in cattle behavior that generated levels of downwind dust lower (P = 0.04) than the levels generated by the behavior resulting from the other feeding pattern. However, the Andersen sampler results showed that there were no differences between feeding patterns with regard to the total number or diversity of microorganisms. The disparity may have been due to the different operating principles of the two systems. The overall numbers of microorganisms recovered were lower than those reported in studies of intensively housed farm animals in which similar recovery techniques were used.


Assembly of amber fragments of the beta subunit of Escherichia coli RNA polymerase.
Immunoblotting of size-separated whole cell proteins permitted the study of protein-protein interaction. Briefly, proteins obtained from cleared cell lysates of Escherichia coli were separated by glycerol gradient centrifugation and analysed by blotting against a set of specific antibodies. We have applied this procedure to the assembly of 11 N-terminal amber fragments of the beta subunit of E. coli RNA polymerase ranging in size between 97% and 23% the length of the intact beta polypeptide (1342 amino acids). In this way, we have been able to define regions on the beta polypeptide involved in the assembly of RNA polymerase.


Competence Mutant of Haemophilus influenzae with Abnormal Ratios of Marker Efficiencies in Transformation.
In studies of competence-deficient mutants of Haemophilus influenzae which absorb deoxyribonucleic acid (DNA) but fail to produce transformants, it was observed that in some mutants the residual transforming activity for different markers varied widely, i.e., produced a ratio effect. One of these mutants, com(-56), was studied intensively to determine the cause of the residual efficiency of transformation and the reason for the ratio effect. The residual frequency of transformation was higher for markers considered single-site mutations (like naladixic acid resistance), whereas the least efficient markers tested were those conferring resistance to high levels of streptomycin or novobiocin which are more complex than single-site mutations. Measurement of frequencies of cotransformation indicated that overall genetic linkage was reduced. Transfection was fairly efficient with phage S2 DNA, but not prophage DNA. Donor marker activity could be detected in transformed cell lysates, but not linked to recipient markers in recombinant molecules. Sucrose gradient analysis of such lysates revealed that donor material was associated with recipient DNA in at least normal quantities, but lacked detectable genetic activity. Material from donor DNA labeled with heavy isotopes was incorporated into recipient chromosomal fragments having a density indistinguishable from normal density, unlike the hybrid density recombinant material found in normal cells. No excessive solubilization or nicking of unincorporated donor was detected. It is postulated that this strain contains a hyperactive nuclease, which reduces the effective size of the input DNA during the integration process.


Studies of apurinic/apyrimidinic endonuclease/ref-1 expression in epithelial ovarian cancer: correlations with tumor progression and platinum resistance.
A crucial step in the DNA base excision repair pathway involves the cleavage of an apurinic/apyrimidinic site by an apurinic/apyrimidinic endonuclease (APE). The major APE in mammalian cells is APE/ref-1, a multifunctional enzyme that acts not only as a DNA repair enzyme but as a redox-modifying factor for a variety of transcription factors. The purpose of this study is to determine whether APE/ref-1 expression differs with ovarian cancer progression and metastasis and in platinum-resistant disease. Ovarian tissue sections were obtained from the Cooperative Human Tissue Network for studies of APE/ref-1 expression and the metastatic process and from our institutional Department of Pathology for studies of APE/ref-1 expression and platinum resistance. Tissue microsections from formalin-fixed, paraffin-embedded specimens of epithelial ovarian cancers were stained using a monoclonal antibody to APE/ref-1 followed by standard immunohistochemical techniques. Slides were then analyzed for the percentage of positively staining nuclei as well as staining intensity using a blinded coding system. All epithelial ovarian cancers expressed APE/ref-1. There were no significant differences in the percentage or intensity of nuclear staining in primary tumors from patients with early- versus advanced-stage disease or in primary tumors versus metastasis from patients with advanced disease. Both platinum-sensitive and platinum-refractory tumors demonstrated a range from minimal to high intensity staining nuclei with a median value of 2+ staining on a scale of 0-3+. The median value for the percentage of nuclei involved was 70% in the platinum-sensitive group and 90% in the platinum-refractory group (P = 0.118). APE/ref-1 expression is ubiquitous among epithelial ovarian cancers and is unaltered with the metastatic process. APE/ref-1 expression does not appear to differ between platinum-sensitive and platinum-refractory ovarian cancers and thus is not a useful biomarker for platinum resistance. Combined with evidence that APE/ref-1 expression and function may not be equivalent in all cell types and tissues, future work will investigate APE/ref-1 as a potential therapeutic target.


Monte Carlo calculations of kQ, the beam quality conversion factor.
To use EGSnrc Monte Carlo simulations to directly calculate beam quality conversion factors, kQ, for 32 cylindrical ionization chambers over a range of beam qualities and to quantify the effect of systematic uncertainties on Monte Carlo calculations of kQ. These factors are required to use the TG-51 or TRS-398 clinical dosimetry protocols for calibrating external radiotherapy beams. Ionization chambers are modeled either from blueprints or manufacturers' user's manuals. The dose-to-air in the chamber is calculated using the EGSnrc user-code egs_chamber using 11 different tabulated clinical photon spectra for the incident beams. The dose to a small volume of water is also calculated in the absence of the chamber at the midpoint of the chamber on its central axis. Using a simple equation, kQ is calculated from these quantities under the assumption that W/e is constant with energy and compared to TG-51 protocol and measured values. Polynomial fits to the Monte Carlo calculated kQ factors as a function of beam quality expressed as %dd(10)x and TPR10(20) are given for each ionization chamber. Differences are explained between Monte Carlo calculated values and values from the TG-51 protocol or calculated using the computer program used for TG-51 calculations. Systematic uncertainties in calculated kQ values are analyzed and amount to a maximum of one standard deviation uncertainty of 0.99% if one assumes that photon cross-section uncertainties are uncorrelated and 0.63% if they are assumed correlated. The largest components of the uncertainty are the constancy of W/e and the uncertainty in the cross-section for photons in water. It is now possible to calculate kQ directly using Monte Carlo simulations. Monte Carlo calculations for most ionization chambers give results which are comparable to TG-51 values. Discrepancies can be explained using individual Monte Carlo calculations of various correction factors which are more accurate than previously used values. For small ionization chambers with central electrodes composed of high-Z materials, the effect of the central electrode is much larger than that for the aluminum electrodes in Farmer chambers.


The role of ethics in pediatrics.
Medical ethics has increased in importance in medical education and practice as a consequence of advances in definitive treatment of patients. Most problems in ethics related to medicine have certain common denominators. Medical scientists and practitioners frequently disclaim any ethical absolutes or consider as important only nonmoral consequences rather than ethical principles such as justice, noninjury, truth-telling, etc. Failure to differentiate moral from nonmoral decisions as well as differences in level of moral reasoning also account for substantial differences in analyzing a given clinical problem. Finally, individual moral policy may be rendered ineffective because of institutional policies that result from purely pragmatic considerations.


Employee acceptance of wearable technology in the workplace.
Wearable technology has many industrial applications. Optimal use adherence and outcomes largely depend on employee acceptance of the technology. This study determined factors that predict employee acceptance of wearables. An online survey of 1273 employed adults asked about demographics, job and organizational characteristics, experience with and beliefs about wearables, and willingness to use wearables. Use cases focused on workplace safety elicited the highest acceptance. An employee's performance expectancy and their organizational safety climate were common predictors of acceptance across use cases. Positive past experiences coincided with involving employees in choosing the device and adequately informing them about data use. Organizations intending to implement wearable technology should (a) focus its use on improving workplace safety, (b) advance a positive safety climate, (c) ensure sufficient evidence to support employees' beliefs that the wearable will meet its objective, and (d) involve and inform employees in the process of selecting and implementing wearable technology.


Identification and Validation of Housekeeping Genes for Gene Expression Analysis of Cancer Stem Cells.
The characterization of cancer stem cell (CSC) subpopulation, through the comparison of the gene expression signature in respect to the native cancer cells, is particularly important for the identification of novel and more effective anticancer strategies. However, CSC have peculiar characteristics in terms of adhesion, growth, and metabolism that possibly implies a different modulation of the expression of the most commonly used housekeeping genes (HKG), like b-actin (ACTB). Although it is crucial to identify which are the most stable HKG genes to normalize the data derived from quantitative Real-Time PCR analysis to obtain robust and consistent results, an exhaustive validation of reference genes in CSC is still missing. Here, we isolated CSC spheres from different musculoskeletal sarcomas and carcinomas as a model to investigate on the stability of the mRNA expression of 15 commonly used HKG, in respect to the native cells. The selected genes were analysed for the variation coefficient and compared using the popular algorithms NormFinder and geNorm to evaluate stability ranking. As a result, we found that: 1) Tata Binding Protein (TBP), Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide (YWHAZ), Peptidylprolyl isomerase A (PPIA), and Hydroxymethylbilane synthase (HMBS) are the most stable HKG for the comparison between CSC and native cells; 2) at least four reference genes should be considered for robust results; 3) the use of ACTB should not be recommended, 4) specific HKG should be considered for studies that are focused only on a specific tumor type, like sarcoma or carcinoma. Our results should be taken in consideration for all the studies of gene expression analysis of CSC, and will substantially contribute for future investigations aimed to identify novel anticancer therapy based on CSC targeting.


Protein-tyrosine phosphatase SHP2 contributes to GDNF neurotrophic activity through direct binding to phospho-Tyr687 in the RET receptor tyrosine kinase.
The signaling mechanisms by which neurotrophic receptors regulate neuronal survival and axonal growth are still incompletely understood. In the receptor tyrosine kinase RET, a receptor for GDNF (glial cell line-derived neurotrophic factor), the functions of the majority of tyrosine residues that become phosphorylated are still unknown. Here we have identified the protein-tyrosine phosphatase SHP2 as a novel direct interactor of RET and the first effector known to bind to phosphorylated Tyr(687) in the juxtamembrane region of the receptor. We show that SHP2 is recruited to RET upon ligand binding in a cooperative fashion, such that both interaction with Tyr(687) and association with components of the Tyr(1062) signaling complex are required for stable recruitment of SHP2 to the receptor. SHP2 recruitment contributes to the ability of RET to activate the PI3K/AKT pathway and promote survival and neurite outgrowth in primary neurons. Furthermore, we find that activation of protein kinase A (PKA) by forskolin reduces the recruitment of SHP2 to RET and negatively affects ligand-mediated neurite outgrowth. In agreement with this, mutation of Ser(696), a known PKA phosphorylation site in RET, enhances SHP2 binding to the receptor and eliminates the effect of forskolin on ligand-induced outgrowth. Together, these findings establish SHP2 as a novel positive regulator of the neurotrophic activities of RET and reveal Tyr(687) as a critical platform for integration of RET and PKA signals. We anticipate that several other phosphotyrosines of unknown function in neuronal receptor tyrosine kinases will also support similar regulatory functions.


Siliplant1 protein precipitates silica in sorghum silica cells.
Silicon is absorbed by plant roots as silicic acid. The acid moves with the transpiration stream to the shoot, and mineralizes as silica. In grasses, leaf epidermal cells called silica cells deposit silica in most of their volume using an unknown biological factor. Using bioinformatics tools, we identified a previously uncharacterized protein in Sorghum bicolor, which we named Siliplant1 (Slp1). Slp1 is a basic protein with seven repeat units rich in proline, lysine, and glutamic acid. We found Slp1 RNA in sorghum immature leaf and immature inflorescence. In leaves, transcription was highest just before the active silicification zone (ASZ). There, Slp1 was localized specifically to developing silica cells, packed inside vesicles and scattered throughout the cytoplasm or near the cell boundary. These vesicles fused with the membrane, releasing their content in the apoplastic space. A short peptide that is repeated five times in Slp1 precipitated silica in vitro at a biologically relevant silicic acid concentration. Transient overexpression of Slp1 in sorghum resulted in ectopic silica deposition in all leaf epidermal cell types. Our results show that Slp1 precipitates silica in sorghum silica cells.


1996 peritoneal dialysis--core indicators report.
The 1996 Peritoneal Dialysis-Core Indicators Study (PD-CIS) retrospectively reviews a random sample of peritoneal dialysis patients from the United States End-Stage Renal Disease (ESRD) program. Peritoneal dialysis (PD) patients are more likely to have a primary diagnosis of glomerulonephritis, less likely to be of African-American heritage, and are younger than hemodialysis patients. One third of PD patients now perform some form of automated peritoneal dialysis (APD) rather than continuous ambulatory peritoneal dialysis (CAPD). The dialysis prescriptions currently employed do not appear to be based on kinetic principles, and the intensity of dialysis achieved is below the proposed minimal guidelines for 30% of patients. In 1996, the mean dialysis index or wKt/Vurea for CAPD patients was 2.0 +/- 0.5 and was not significantly altered from the 1995 value of 2.1. Eighty-four percent of CAPD patients perform four or fewer exchanges daily, and only 27% of patients have prescriptions using infusion volumes greater than 2 L. Although hematocrits have improved since 1995, 30% of PD patients have a hematocrit below 30%. The mean serum albumin for PD patients is 3.5 g/dL, and 25% of patients have a 6-month average serum albumin value below 3.2 g/dL. In general, the indices monitored as predictive of health and well-being of PD patients afford significant opportunity for improvement.


Primum non nocere, safety of continuous renal replacement therapy.
To highlight selected adverse effects of continuous renal replacement therapy and to suggest measures for reducing harm. Adverse effects of continuous renal replacement therapy are related to bioincompatibility, bleeding, metabolic consequences, loss of heat, vascular access and human error. Bioincompatibility initiates coagulation and inflammation. Use of membranes with high pore size permits direct removal of the locally formed inflammatory mediators. Regional anticoagulation with citrate reduces the risk of bleeding and abolishes polymorphonuclear and platelet degranulation in the filter. Continuous renal replacement therapy is associated with increased losses of amino acids and the water-soluble vitamins carnitine and selenium. Compared to bicarbonate, lactate-buffered fluids may increase energy needs and glucose intolerance. Patients on continuous renal replacement therapy may lose heat. Several measures can be considered to improve safety of continuous renal replacement therapy. Among these are the use of membranes with high pore size, implementation of citrate to reduce bleeding and bioincompatibility, supplementation of water-soluble vitamins and selenium to increase antioxidant defense, use of bicarbonate-buffered solutions in the most critically ill, and prevention of loss of heat. Last but not least, improvement of safety needs continuous training of staff for better understanding of the method.


The prognostic value of E-cadherin and the cadherin-associated molecules alpha-, beta-, gamma-catenin and p120ctn in prostate cancer specific survival: a long-term follow-up study.
To determine the value of loss of expression of E-cadherin and cadherin associated molecules as prognostic markers for prostate cancer patients in a long-term follow-up study. Sixty-five prostate cancer specimens, obtained from patients with different stages of prostate cancer who underwent a radical prostatectomy or TUR-P between 1987 and 1991, were used for immunohistochemical analysis of the expression pattern of E-cadherin, alpha-, beta-, gamma-catenin and p120(ctn). Clinical records of these patients were studied for follow-up data and the prognostic value of expression of these adhesion molecules was determined by Kaplan-Meier survival analyses and multivariable proportional hazard regression analysis. Normal staining patterns were found in 36 cases (55.4%) for E-cadherin, 37 cases (56.9%) for alpha-catenin, 40 cases (61.5%) for beta-catenin, 25 cases (38.5%) for gamma-catenin, and 40 cases (61.5%) for p120(ctn). Overall, a strong correlation was found between the expression of E-cadherin and other cadherin-associated molecules. The 5-year survival rates for each staining were as follows: E-cadherin (normal 79.2%, aberrant 26.8%), alpha-catenin (normal 79.2%, aberrant 26.8%), beta-catenin (normal 73.1%, aberrant 27.3%), gamma-catenin (normal 86.4%, aberrant 37.1%), and p120(ctn) (normal 72.8%, aberrant 30.0%). There was a significant difference in survival between normal and aberrant expression in each staining (log rank P < 0.0001). The proportional hazard regression model including tumor stage and Gleason score revealed alpha-catenin expression as the best prognostic marker for patients with prostate cancer. Our data revealed a strong correlation between E-cadherin expression and other cadherin-associated molecules. Among these markers, alpha-catenin seems the best prognostic marker for prostate cancer specific survival. Larger studies are needed to confirm this result.


Yeast Deletomics to Uncover Gadolinium Toxicity Targets and Resistance Mechanisms.
Among the rare earth elements (REEs), a crucial group of metals for high-technologies. Gadolinium (Gd) is the only REE intentionally injected to human patients. The use of Gd-based contrasting agents for magnetic resonance imaging (MRI) is the primary route for Gd direct exposure and accumulation in humans. Consequently, aquatic environments are increasingly exposed to Gd due to its excretion through the urinary tract of patients following an MRI examination. The increasing number of reports mentioning Gd toxicity, notably originating from medical applications of Gd, necessitates an improved risk-benefit assessment of Gd utilizations. To go beyond toxicological studies, unravelling the mechanistic impact of Gd on humans and the ecosystem requires the use of genome-wide approaches. We used functional deletomics, a robust method relying on the screening of a knock-out mutant library of Saccharomyces cerevisiae exposed to toxic concentrations of Gd. The analysis of Gd-resistant and -sensitive mutants highlighted the cell wall, endosomes and the vacuolar compartment as cellular hotspots involved in the Gd response. Furthermore, we identified endocytosis and vesicular trafficking pathways (ESCRT) as well as sphingolipids homeostasis as playing pivotal roles mediating Gd toxicity. Finally, tens of yeast genes with human orthologs linked to renal dysfunction were identified as Gd-responsive. Therefore, the molecular and cellular pathways involved in Gd toxicity and detoxification uncovered in this study underline the pleotropic consequences of the increasing exposure to this strategic metal.


[The clinical picture of psychoneurotic disorders in neurodermatitis, psoriasis and alopecia totalis in children].
Clinical, psychological, and electroencephalographic examinations of 267 children suffering from neurodermatitis, psoriasis, and total alopecia have revealed neural and mental disorders in 73.78% of patients, these disorders being the most grave and polymorphic in neurodermatitis.


Identification and expression of microRNA in the brain of hibernating bats, Myotis lucifugus.
Recent research has highlighted roles for non-coding RNA i7n the regulation of stress tolerance in bats. In this study, we propose that microRNA could also play an important role in neuronal maintenance during hibernation. To explore this possibility, RT-PCR was employed to investigate the expression of eleven microRNAs from the brain tissue of euthermic control and torpid bats. Results show that eight microRNAs (miR-21, -29b, -103, -107, -124a, -132, -183 and -501) increased (1.2-1.9 fold) in torpid bats, while the protein expression of Dicer, a microRNA processing enzyme, did not significantly change during torpor. Bioinformatic analysis of the differentially expressed microRNA suggests that these microRNAs are mainly involved in two processes: (1) focal adhesion and (2) axon guidance. To determine the extent of microRNA sequence conservation in the bat, we successfully identified bat microRNA from sequence alignments against known mouse (Mus musculus) microRNA. We successfully identified 206 conserved pre-microRNA sequences, leading to the identification of 344 conserved mature microRNA sequences. Sequence homology of the identified sequences was found to be 94.76±3.95% and 98.87±2.24% for both pre- and mature microRNAs, respectively. Results suggest that brain function related to the differentiation of neurons and adaptive neuroprotection may be under microRNA control during bat hibernation.


Morphine dependence and diabetes. II. Alterations of normorphine potency in the guinea-pig ileum and mouse vas deferens and of ileal morphine dependence by changes in glucose concentration.
The effects of glucose in vitro at concentrations of 100, 200 and 400 mg/dl on responses of the guinea-pig ileum and mouse vas deferens to opiates were studied. Responses of the electrically stimulated ileum to normorphine were significantly reduced (IC50 values: 26.3 +/- 4.3, 40.5 +/- 3.7 and 71.8 +/- 16.6 nM) as the glucose concentration increased, whereas the potency of normorphine was not affected by equimolar substitutions of glucose with the nonmetabolizable sugar 3-O-methylglucose. The potency of naloxone to induce withdrawal contractions of ilea incubated in Ringer's solution for 3 or 4.5 hr with 1 microM morphine was also significantly decreased as the concentration of glucose in the incubations increased, indicating a reduction of acute dependence development by high glucose. The potency of normorphine was also reduced by increasing concentrations of glucose in electrically stimulated vasa deferentia from nondiabetic (db/m+, m+/m+) C57BL/KsJ mice (IC50 values: 0.53 +/- 0.01, 0.77 +/- 0.01 and 1.84 +/- 0.03 nM), but not in vasa deferentia from their diabetic (db/db) littermates. These results support our hypothesis that the decreased sensitivity to morphine and decreased development of physical dependence on morphine in in vivo models of diabetes reported previously by our laboratory is due primarily to the hyperglycemia associated with diabetes.


[Taxonomic position of certain representatives of sulphate-reducing corrosive microbial community].
Sulfate-reducing bacteria and their heterotrophic satellites have been isolated from the biofilm formed on steel by sulfidogenic corrosive microbial community. Bacteria were characterized according to phenotypical features and investigated by the methods of molecular-genetic analysis. In accordance with the phenotypical features the studied strain of sulfate-reducing bacteria were related to Desulfovibrio genus, Gram-positive strains of heterotrophic satellites were related to Bacillus genus, B. firmus and B. subtilis species. Gram-negative strains, as to their physiological-biochemical characteristics were related to Pseudomonas genus, P. aeruginosa species, Stenotrophomonas genus, S. maltophilia species, and Aeromonas genus, A. hydrophila/caviae species. Taxonomic position of certain representatives of the community is confirmed by the molecular-genetic methods. A comparative analysis of the sequencing results has evidenced for the identity of sequences of 16S rRNA of the studied bacteria with analogous sequence of strains from the GenBank database. Nucleotide sequence of strain 27 has a 99% homology with the sequence of Pseudomonas aeruginosa, strain 36 deposited at GenBank to nucleotide sequence of Bacillus subtilis. As to sulfate-reducing bacteria, only their belonging to Desulfovibrio genus has been confirmed. Thus the results of sequence-analysis of 16S rRNA genes are in agreement with the data obtained from studying the phenotypical features.


Inter-professional practice in the prevention and management of child and adolescent self-harm: foster carers' and residential carers' negotiation of expertise and professional identity.
Inter-professional collaboration remains a significant concern within healthcare and social care. However, there has been scant attention paid to practices at the interface of clinicians and carers, namely foster carers and residential carers. The present study considers child and adolescent self-harm management and prevention practices as a site of empirical interest due to reports that multi-agency teams are not effectively operating. Drawing upon a grounded theory approach, data were generated via semi-structured interviews and focus groups with residential carers (n = 15) and foster carers (n = 15) in Wales. Themes were developed through axial coding. The results present two central themes to explain the nature and perceived causes of inter-professional discord. First, there are clear contestations in expertise, with carers challenging clinicians' propositional knowledge in favour of their own experiential expertise. However, participants simultaneously endorse medical dominance, which contributes to their sense of disempowerment and marginalisation. Second, is the preclusion of carers' professional identity, primarily due to inadequate professionalisation procedures. Meanwhile, the privileging of their parenting role is perceived to support the perpetuation of courtesy stigma. Carers are then compelled to undertake the effortful labour of legitimisation. Together these thematic insights provide direction on mechanisms to improve inter-professional interactions, notably around training and accreditation.


Forefoot reconstruction by reversed island flaps in diabetic patients.
Soft-tissue coverage of the foot is often difficult, especially when the distal third of the foot (dorsal or plantar aspects) is involved. The clinical situation can be further complicated when diabetic patients are affected by painful and unstable wounds of this kind because of the familiar phenomenon of vasculopathy. The purpose of this study was to evaluate the possibility of using distally based foot flaps to cover forefoot defects in diabetic patients. Preoperative selection of patients was the key to this study; those who had other major disease, chronic infection, bone involvement, and/or insufficient foot vascularization were excluded from the study. The authors report a series of 12 diabetic patients in whom the reconstruction of medium-sized defects (ranging from 1.5 x 2.0 cm to 3.0 x 7.0 cm) of the forefoot was performed using distally based dorsalis pedis flaps or medial plantar flaps. The transferred flaps survived and adapted well to the defects, except for one flap in a patient who had a slight venous insufficiency at outset. Wearing their own footwear, patients could walk after 20 to 30 days. After the follow-up period (3 months to 3 years), no skin breakdown in the treated areas was observed. Temporary donor-site pain was reported by medial plantar flap patients, and partial skin graft loss at the donor site occurred in some of the dorsalis pedis patients. The authors suggest that in selected cases, medium-sized soft-tissue defects involving the dorsal aspects or the weight-bearing areas of the diabetic foot can be successfully covered with distally based island flaps.


[A not very essential obesity: the Rohhad syndrome. Description of two cases and review of the literature].
Rapid-onset Obesity with Hypothalamic Dysfunction, Hypoventilation, and Autonomic Dysregulation (ROHHAD) is a rare and complex pediatric disorder. Children typically show ROHHAD after the first years of life with rapid weight gain and subsequently autonomic nervous system dysregulation (altered pain perception, pupillary dysfunction, hypothermia and bradycardia); alveolar hypoventilation with risk of cardiorespiratory arrest and hypothalamic dysfunction (central diabetes insipidus, hypothyroidism, growth hormone and corticotrophin deficiency). Tumours of neural crest origin, such as ganglioneuroblastoma and ganglioneuronoma, are reported in 33% of the patients and may be found in the chest or abdomen. Here we describe two girls who presented with rapid weight gain, at the age of 5 and 9 years respectively. The first was admitted due to obesity and central hypothyroidism. After two months she rapidly developed a clinical picture characterized by thermal dysregulation, hypodipsia and severe hypernatriemia, hypertrigliceridemia, alveolar hypoventilation supported by mechanical ventilation. The second presented with rapid-onset obesity and a mild hyperprolactinemia. After three months of follow-up she was admitted due to a clinical picture of hypothermia, seizures and hyponatremia. Subsequentely she developed altered water balance (severe hypernatremia) and severe hypoventilation. Chest CT and MR imaging showed a posterior mediastinal mass. Endocrinological investigation showed corticotrophin deficiency and central hypothyroidism treated with specific replacement therapies. On the basis of our experiences we can infer that it is necessary perform specific further investigations of hypothalamic function in all the children with rapid onset obesity in order to early prevent the catastrophic consequences that may occur in this syndrome.


Underwater turning movement during foraging in Hydromedusa maximiliani (Testudines, Chelidae) from southeastern Brazil.
A type of locomotor behavior observed in animals with rigid bodies, that can be found in many animals with exoskeletons, shells, or other forms of body armor, to change direction, is the turning behavior. Aquatic floated-turning behavior among rigid bodies animals have been studied in whirligig beetles, boxfish, and more recently in freshwater turtle, Chrysemys picta. In the laboratory we observed a different kind of turning movement that consists in an underwater turning movement during foraging, wherein the animal pivoted its body, using one of the hindlimbs as the fixed-point support in the substratum. We describe, analyze and quantify this movement during foraging in Hydromedusa maximiliani, using observations made in the laboratory. We studied 3 adult specimens (2 males, 1 female) and 2 non-sexed juveniles of H. maximiliani. They were kept individually in an aquarium filled with water and small fish. They were filmed, in dorsal view, at 30 frames per second. Sequences were analyzed frame by frame and points were marked on limbs and shell to enable analysis of variation in limb flexion and extension, as well as rotation movements. While foraging, turtles frequently turned their bodies, using one hind leg as the pivot point. This underwater turning movement, in addition to slow movements with the neck stretched, or staying nearly immobile and scanning the surroundings with lateral movements of the neck (in arcs up to 180°), and fast attacks of neck, may increase prey capture rates.


Analysis of 15 novel full-length BK virus sequences from three individuals: evidence of a high intra-strain genetic diversity.
To determine the variability of BK virus (BKV) in vivo, the sequences of nine full-length molecular clones from the striated muscle and heart DNA of a patient with BKV-associated capillary leak syndrome (BKVCAP), as well as three clones each from the urine of one human immunodeficiency virus type 2-positive (BKVHI) and one healthy control subject (BKVHC), were analysed. The regulatory region of all clones corresponded to the archetypal regulatory region usually found in urine isolates. Analysis of the predicted conformation of BKVCAP proteins did not suggest any structural differences on the surface of the viral particles compared with BKVHI and BKVHC clones. No amino acid changes common to most BKVCAP clones could be identified that have not already been reported in non-vasculotropic strains. However, the coding region of each clone had unique nucleotide substitutions, and intra-host variability was greater among BKVCAP clones, with a mean difference of 0.29 % per site compared with 0.16 % for BKVHI and 0.14 % for BKVHC. The clones from each strain formed monophyletic clades, suggesting a single source of infection for each subject. The most divergent BKVCAP clones differed at 0.55 % of sites, implying a rate of nucleotide substitution of approximately 5 x 10(-5) substitutions per site per year, which is two orders of magnitude faster than estimated for the other human polyomavirus, JC virus.


Use of hydrophobic moment plot methodology to aid the identification of oblique orientated alpha-helices.
A number of alpha-helix forming peptides have been reported which appear to promote membrane fusion and other biological events related to the disruption of a hydrophobic/hydrophilic interface, due to the presence of a hydrophobicity gradient along the helical long axis. When alpha-helices from this class were analysed according to hydrophobic moment plot methodology a linear association was found to exist between the mean hydrophobic moment, <microH>, and the corresponding mean hydrophobicity, <H0>. This association was described by the least squares regression line: <microH>=0.508-0.422<H0> and, here, a methodology to aid the prediction of oblique orientated alpha-helices is presented, based on a 99% prediction band around this regression line. This methodology is intended to provide an initial identification of candidates for further investigation by other techniques such as the molecular hydrophobic potential and laboratory based experimentation, not to assign function.


The effects of estrogen and raloxifene treatment on antioxidant enzymes in brain and liver of ovarectomized female rats.
Recent studies documented that estrogen have antioxidant properties in-vitro, there are conflicting results on the effect of estrogen in vivo. We aimed to investigate the effects of estradiol and Raloxifene on the antioxidant enzyme [superoxide dismutase (SOD) and catalase (CAT)] activities and MDA levels in brain and liver homogenates of ovariectomized female rats. Twelve weeks after ovariectomy, female Sprague-Dawley rats (n = 26) were divided into three groups: (1) Ovariectomized placebo group (n = 6) was given physiologic saline. (2) Estrogen group (n = 10) was given Ethynyl estradiol, 0.1 mg/kg sc. (3) Raloxifene group (n = 10) was given raloxifene, 1 mg/kg sc during 8 weeks. Ten rats were used as naive controls without any treatment (Sham operated group, n = 10). Ovariectomy lead to an increase in the CAT activities in liver tissue samples compared to the sham group (p = 0.056, Mann-Whitney test). While estrogen treatment reversed to normal levels of CAT activities, raloxifene remained as ineffective. Superoxide dismutase activities and MDA levels in liver were remained unchanged in all groups. There was no significant change in the brain tissue SOD and CAT activities between the control ovariectomy, estrogen treated, and raloxifen treated groups. We determined an increase in MDA levels in brain of ovariectmised rat (p = 0.02). While raloxifene treatment reversed to normal levels of MDA (p = estrogen treatment failed. Our data showed that estrogen may play a role in regulation of CAT and SOD activities in liver due to its antioxidative effects. We can suggest estrogen and raloxifene exert their antioxidative effects in brain rather than liver. Since Raloxifene's effect is more clear than estradiol, raloxifene may be suggested primarily for treatment and/or prevention of diseases which can be resulted from oxidative stress in postmenopausal women.


Adsorption of avidin on microfabricated surfaces for protein biochip applications.
The adsorption of the protein avidin from hen egg white on patterns of silicon dioxide and platinum surfaces on a microchip and the use of fluorescent microscopy to detect binding of biotin are described. A silicon dioxide microchip was formed using plasma-enhanced chemical vapor deposition while platinum was deposited using radiofrequency sputtering. After cleaning using a plasma arc, the chips were placed into solutions containing avidin or bovine serum albumin. The avidin was adsorbed onto the microchips from phosphate-buffered saline (PBS) or from PBS to which ammonium sulfate had been added. Avidin was also adsorbed onto bovine serum albumin (BSA)-coated surfaces of oxide and platinum. Fluorescence microscopy was used to confirm adsorption of labeled protein, or the binding of fluorescently labeled biotin onto previously adsorbed, unlabeled avidin. When labeled biotin in PBS was presented to avidin adsorbed onto a BSA-coated microchip, the fluorescence signal was significantly higher than for avidin adsorbed onto the biochip alone. The results show that a simple, low-cost adsorption process can deposit active protein onto a chip in an approach that has potential application in the development of protein biochips for the detection of biological species.


The acute effects of dimebolin, a potential Alzheimer's disease treatment, on working memory in rhesus monkeys.
Dimebolin (latrepirdine), a compound with multiple potential drug targets, is being evaluated in clinical trials for the treatment of Alzheimer's disease (AD) and preliminary results suggest it can slow the disease process. There is also evidence that dimebolin directly improves aspects of cognition. Here we examined the acute effect of dimebolin on components of working memory in non-human primates, young adult (11-17 years old) and aged (20-31 years old) rhesus macaques. The effects of dimebolin (3.9-118 µg kg(-1)) on working memory, as measured by performance on delayed matching-to-sample (DMTS), were examined in the normal young adult monkeys and aged adult monkeys. All the monkeys studied were proficient in the performance of a computer-assisted DMTS task. In a subsequent experiment in the same subjects, dimebolin was administered 15 min before a cognitively-impairing dose (20 µg kg(-1)) of scopolamine. In both the young adult and aged monkeys, dimebolin significantly increased the DMTS task accuracies. In young adults, the task improvement was associated with long (retention/retrieval) delay trials, and a protracted enhancement was observed for sessions run 24 h post administration of a single dose. Dimebolin did not significantly attenuate the scopolamine-induced impairment. In the aged monkeys, dimebolin significantly improved the reduced task accuracies associated with long delay intervals. Here we demonstrated that dimebolin is able to improve components of working memory in monkeys and to induce a protracted response for at least 24 h after administration of a single dose.


The Septin Gene StSep4 Contributes to the Pathogenicity of Setosphaeria turcica by Regulating the Morphology, Cell Wall Integrity, and Pathogenic Factor Biosynthesis.
Septins are a conserved group of GTP-binding proteins found in all eukaryotes and are the fourth-most abundant cytoskeletal proteins. Septins of some pathogenic fungi are involved in morphological changes related to infection. Our previous studies have identified four core septins (StSep1-4) in Setosphaeria turcica, the causal agent of northern corn leaf blight, while only StSep4 is significantly upregulated during the invasive process. We therefore used forchlorfenuron (FCF), the specific inhibitor of septin, and ΔStSep4 knockout mutants to further clarify the role of septins in S. turcica pathogenicity. FCF treatment caused a dose-dependent reduction in S. turcica colony growth, delayed the formation of infection structures, and reduced the penetration ability. ΔStSep4 knockout mutants displayed abnormal mycelium morphology, slow mycelial growth, conidiation deficiency, delayed appressorium development, and weakened pathogenicity. StSep4 deletion also broke cell wall integrity, altered chitin distribution, decreased the melanin content, and disrupted normal nuclear localization. A transcriptomic comparison revealed that genes differentially expressed between ΔStSep4 and WT were enriched in terms of ribosomes, protein translation, membrane components, and transmembrane transport activities. Our results demonstrate that StSep4 is required for morphology and pathogenicity in S. turcica, making it a promising target for the development of novel fungicides.


Status epilepticus of inflammatory etiology: a cohort study.
Inflammation-related epilepsy is increasingly recognized; however, studies on status epilepticus (SE) are very infrequent. We therefore aimed to determine the frequency of inflammatory etiologies in adult SE, and to assess related demographic features and outcomes. This was a retrospective analysis of a prospective registry of adult patients with SE treated in our center, from January 2008 to June 2014, excluding postanoxic causes. We classified SE episodes into 3 etiologic categories: infectious, autoimmune, and noninflammatory. Demographic and clinical variables were analyzed regarding their relationship to etiologies and functional outcome. Among the 570 SE consecutive episodes, 33 (6%) were inflammatory (2.5% autoimmune; 3.3% infectious), without any change in frequency over the study period. Inflammatory SE episodes involved younger patients (mean age 53 vs 61 years, p = 0.015) and were more often refractory to initial antiepileptic treatment (58% vs 38%, odds ratio = 2.19, 95% confidence interval = 1.07-4.47, p = 0.041), despite similar clinical outcome. Subgroup analysis showed that, compared with infectious SE episodes, autoimmune SE involved younger adults (mean age 44 vs 60 years, p = 0.017) and was associated with lower morbidity (return to baseline conditions in 71% vs 32%, odds ratio = 5.41, 95% confidence interval = 1.19-24.52, p = 0.043) without any difference in mortality. Despite increasing awareness, inflammatory SE etiologies were relatively rare; their occurrence in younger individuals and higher refractoriness to treatment did not have any effect on outcome. Autoimmune SE episodes also occurred in younger patients, but tended to have better outcomes in survivors than infectious SE.


[The vagus-inhibiting action of alpha-endorphin].
An intravenous bolus injection of 0.1 ml alpha-endorphin (1 x 10(-8)-1 x 10(-4) g/ml) didn't change the heart rate in frogs. The parasympathetic bradycardia induced by the peripheral vagus stimulation was decreased by alpha-endorphin. This vago-inhibitory action was dose-dependent (1 x 10(-5)-1 x 10(-4) g/ml). The maximal inhibitory action was watched in 4-8 and 9-15 minutes after bolus injection of alpha-endorphin in concentration of 1 x 10(-5) and 1 x 10(-4) g/ml accordingly.