2018 updated European League Against Rheumatism evidence-based recommendations for the diagnosis of gout.
Although gout is the most common inflammatory arthritis, it is still frequently misdiagnosed. New data on imaging and clinical diagnosis have become available since the first EULAR recommendations for the diagnosis of gout in 2006. This prompted a systematic review and update of the 2006 recommendations. A systematic review of the literature concerning all aspects of gout diagnosis was performed. Recommendations were formulated using a Delphi consensus approach. Eight key recommendations were generated. A search for crystals in synovial fluid or tophus aspirates is recommended in every person with suspected gout, because demonstration of monosodium urate (MSU) crystals allows a definite diagnosis of gout. There was consensus that a number of suggestive clinical features support a clinical diagnosis of gout. These are monoarticular involvement of a foot or ankle joint (especially the first metatarsophalangeal joint); previous episodes of similar acute arthritis; rapid onset of severe pain and swelling; erythema; male gender and associated cardiovascular diseases and hyperuricaemia. When crystal identification is not possible, it is recommended that any atypical presentation should be investigated by imaging, in particular with ultrasound to seek features suggestive of MSU crystal deposition (double contour sign and tophi). There was consensus that a diagnosis of gout should not be based on the presence of hyperuricaemia alone. There was also a strong recommendation that all people with gout should be systematically assessed for presence of associated comorbidities and risk factors for cardiovascular disease, as well as for risk factors for chronic hyperuricaemia. Eight updated, evidence-based, expert consensus recommendations for the diagnosis of gout are proposed.


Multiple receptors for calcitonin gene-related peptide and amylin on guinea-pig ileum and vas deferens.
1. The responses of the electrically stimulated guinea-pig ileum and vas deferens to human and rat calcitonin gene-related peptide (CGRP) and amylin were investigated. 2. The inhibition of contraction of the ileum produced by human alpha CGRP was antagonized by human alpha CGRP8-37 (apparent pA2 estimated at 7.15 +/- 0.23) > human alpha CGRP19-37 (apparent pA2 estimated as 6.67 +/- 0.33) > [Tyr0]-human alpha CGRP28-37. The amylin antagonist, AC187, was three fold less potent than CGRP8-37 in antagonizing human alpha CGRP. 3. Both human beta- and rat alpha CGRP inhibited contractions of the ileum, but this was less sensitive to inhibition by CGRP8-37 than the effect of human alpha CGRP. However, CGRP19-37 was twenty times more effective in inhibiting the response to rat alpha CGRP (apparent pA2 estimated as 8.0 +/- 0.1) compared to human alpha CGRP. 4. Rat amylin inhibited contractions in about 10% of ileal preparations; this effect was not antagonized by any CGRP fragment. Human amylin had no action on this preparation. 5. Both human and rat alpha CGRP inhibited electrically stimulated contractions of the vas deferens, which were not antagonized by 3 microM CGRP8-37 or 10 microM AC187. 6. Rat amylin inhibited the stimulated contractions of the vas deferens (EC50 = 77 +/- 9 nM); human amylin was less potent (EC50 = 213 +/- 22 nM). The response to rat amylin was antagonized by 10 microM CGRP8-37 (EC50 = 242 +/- 25 nM) and 10 microM AC187 (EC50 = 610 +/- 22 nM). 7. It is concluded that human alpha CGRP relaxes the guinea-pig ileum via CGRP1-like receptors, but that human beta CGRP and rat alpha CGRP may use additional receptors. These are distinct CGRP2-like and amylin receptors on guinea-pig vas deferens.


Platyzoma: a new look at an old link in ferns.
Record of a chromosome number of 2n = 76, unusual for pteridophytes, in Platyzoma microphyllum R. Br. emphasizes other unique features of this monotypic Queensland fern and provides new evidence of its possible relationships. Other characteristics of this plant, which are not known among terrestrial ferns, are incipient heterospory-having two sizes of spores and a dioecious condition of the gametophytes. These and other morphological features show relationships of Platyzoma to members of the Schizaeaceae and Marsileaceae and relationships of these families to the Polypodiaceae, in which it is treated under the subfamily Platyzomatoideae.


Modifications to ART service delivery models by health facilities in Uganda in promotion of intervention sustainability: a mixed methods study.
In November 2015, WHO released new treatment guidelines recommending that all diagnosed as HIV positive be enrolled on antiretroviral therapy (ART). Sustaining and expanding ART scale-up programs in resource-limited settings will require adaptations and modifications to traditional ART delivery models to meet the rapid increase in demand. We identify modifications to ART service delivery models by health facilities in Uganda to sustain ART interventions over a 10-year period (2004-2014). A mixed methods approach involving two study phases was adopted. In the first phase, a survey of a nationally representative sample of health facilities (n = 195) in Uganda which were accredited to provide ART between 2004 and 2009 was conducted. The second phase involved semi-structured interviews (n = 18) with ART clinic managers of 6 of the 195 health facilities purposively selected from the first study phase. We adopted a thematic framework consisting of four categories of modifications (format, setting, personnel, and population). The majority of health facilities 185 (95%) reported making modifications to ART interventions between 2004 and 2014. Of the 195 health facilities, 157 (81%) rated the modifications made to ART as "major." Modifications to ART were reported under all the four themes. The quantitative and qualitative findings are integrated and presented under four themes. Format: Reducing the frequency of clinic appointments and pharmacy-only refill programs was identified as important strategies for decongesting ART clinics. Home-based care programs were introduced to reduce provider ART delivery costs. Personnel: Task shifting to non-physician cadre was reported in 181 (93%) of the health facilities. Visits to the ART clinic were rationalized in favor of the sub-population deemed to have more clinical need. Two health facilities focused on patients living nearer the health facilities to align with targets set by external donors. Over the study period, health facilities made several modifications ART interventions to improve fit with their resource-constrained settings thereby promoting long-term sustainability. Further research evaluating the effect of these modifications on patient outcomes and ART delivery costs is recommended. Our findings have implications for the sustainability of ART scale-up programs in Uganda and other resource-limited settings.


Integrated biochemical and computational approach identifies BCL6 direct target genes controlling multiple pathways in normal germinal center B cells.
BCL6 is a transcriptional repressor required for mature B-cell germinal center (GC) formation and implicated in lymphomagenesis. BCL6's physiologic function is only partially known because the complete set of its targets in GC B cells has not been identified. To address this issue, we used an integrated biochemical-computational-functional approach to identify BCL6 direct targets in normal GC B cells. This approach includes (1) identification of BCL6-bound promoters by genome-wide chromatin immunoprecipitation, (2) inference of transcriptional relationships by the use of a regulatory network reverse engineering approach (ARACNe), and (3) validation of physiologic relevance of the candidate targets down-regulated in GC B cells. Our approach demonstrated that a large set of promoters (> 4000) is physically bound by BCL6 but that only a fraction of them is repressed in GC B cells. This set of 1207 targets identifies several cellular functions directly controlled by BCL6 during GC development, including activation, survival, DNA-damage response, cell cycle arrest, cytokine signaling, Toll-like receptor signaling, and differentiation. These results define a broad role of BCL6 in preventing centroblasts from responding to signals leading to exit from the GC before they complete the phase of proliferative expansion and of antibody affinity maturation.


Expression and dimerization of the rat activin subunits betaC and betaE: evidence for the ormation of novel activin dimers.
Activins are cytokines of the transforming growth factor beta family, which plays a central role in the determination of cell fate and the regulation of tissue balance. Family members are composed of two subunits and this dimerization is critical for liganding their cognate receptors and execution of proper functions. In the current study we focused on the localization of activin betaA, betaB, betaC and betaE subunits in the adult rat and analyzed the composition of putative activin beta dimers. By dissecting tissue distribution of various activins, we found that the liver, in particular the hepatocytes, is the major source for activin betaC and betaE transcripts, since other tissues almost failed to express these isoforms. In sharp contrast, the emergence of activin betaA and betaB appeared ubiquitous. Using a highly selective proteome approach, we were able to identify homo- as well as heterodimers of individual activin subunits, indicating a high redundancy of ligand composition. Certainly, this broad potential to homo- and heterodimerize has to be considered in future studies on activin function.


Virus-like particles as vaccines and vessels for the delivery of small molecules.
Virus-like particles (VLPs) structurally mimic the viral capsid and have therefore been extensively, and quite successfully, used as vaccine and viral serology reagents. The ability of VLPs to include nucleic acids and small molecules has also made them novel vessels for gene and drug delivery. The regular, repetitive surface of VLPs has been exploited as a template for nanoscale synthesis. Recent progress has been made in the development of several virus models.


[Supramolecular organization of glycolytic enzymes].
On the basis of the analysis of the data on adsorption of glycolytic enzymes to structural proteins of skeletal muscle and to erythrocyte membranes, the data on enzyme-enzyme interactions and the data on the regulation of activity of glycolytic enzymes by cellular metabolites the structure of glycolytic enzyme complex adsorbed to a biological support has been proposed. The key role in the formation of the multienzyme complex belongs to 6-phosphofructokinase. The enzyme molecule has two association sites, one of which provides the fixation of 6-phosphofructokinase on the support and another is saturated by fructose-1,6-bisphosphate aldolase. The multienzyme complex fixed on structural proteins of skeletal muscle contains one tetrameric molecule of 6-phosphofructokinase and at two molecules of other glycolytic enzymes. Hexokinase is not involved in the complex composition. The molecular mass of the multienzyme complex is about 2,6 X 10(6) Da. The formation of the multienzyme complex leads to the compartmentation of the glycolytic process. The problem of integration of physico-chemical mechanisms of enzyme activity regulation (allosteric, dissociative and adsorptive mechanisms) is discussed.


THE EFFECT OF ANTISERUM, ALONE AND WITH HYDROCORTISONE, ON FOETAL MOUSE BONES IN CULTURE.
1. The effects of normal rabbit serum and of rabbit antiserum to whole foetal mouse tissues, on the isolated limb bones of late foetal mice were studied in organ culture, and the influence of hydrocortisone on these effects was investigated. 2. Unheated normal serum caused slight loss of metachromatic material from the cartilage matrix, and some resorption of both cartilage and bone. 3. In unheated antiserum to foetal mouse tissues, the terminal cartilage was smaller and less metachromatic than in paired controls in normal serum, while osteoclasis was so intense that in many explants the bone had almost disappeared. The amount of necrosis varied with different batches of antiserum. 4. The changes produced by normal serum and antiserum could be largely prevented by heating the sera to 57 degrees C for 45 minutes. 5. The effects could also be inhibited by the addition of hydrocortisone to the unheated sera; as little as 0.1 microg hydrocortisone per ml of medium had a well marked protective action. 6. It is suggested that (a) unheated antiserum causes a release of lysosomal enzymes with consequent breakdown of intercellular material, (b) this release is due to an indirect action on the lysosome via an increased permeability of the cell membrane, (c) hydrocortisone does not affect the antigen-antibody reaction, but inhibits the autolytic changes that normally follow this reaction, possibly by stabilising both the lysosomal and cell membranes.


Effect of Vitamin D3 Supplementation on Respiratory Tract Infections in Healthy Individuals: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Vitamin D supplementation may be a simple preventive measure against respiratory tract infections (RTIs) but evidence from randomized controlled trials is inconclusive. We aimed to systematically summarize results from interventions studying the protective effect of vitamin D supplementation on clinical and laboratory confirmed RTIs in healthy adults and children. Medline, EMBASE, CENTRAL, and CINAHL were screened from inception until present (last updated in January 2016) completed by a search of the grey literature, clinical trial registers and conference abstracts. We included randomized trials comparing vitamin D versus placebo or no treatment. Two independent reviewers were responsible for study selection and data extraction. Cochrane's risk of bias tool and the GRADE approach were used for quality assessment. Estimates were pooled with random-effects models. Heterogeneity was explored by sub-group and meta-regression analyses. Of 2627 original hits, 15 trials including 7053 individuals were ultimately eligible. All used oral cholecalciferol. We found a 6% risk reduction with vitamin D3 supplementation on clinical RTIs, but the result was not statistically significant (RR 0.94; 95% CI 0.88 to 1.00). Heterogeneity was large (I-square 57%) and overall study quality was low. There were too few studies to reliably assess a potential risk reduction of laboratory confirmed RTI. Evidence was insufficient to demonstrate an association between vitamin D supplementation and risk of clinical RTI in sub-groups with vitamin D deficiency. In previously healthy individuals vitamin D supplementation does not reduce the risk of clinical RTIs. However, this conclusion is based on a meta-analysis where the included studies differed with respect to population, baseline vitamin D levels and study length. This needs to be considered when interpreting the results. Future trials should focus on vitamin D deficient individuals and apply more objective and standardized outcome measurements.


Radioimmunoassay of interleukin-6 in plasma.
We present a double-antibody radioimmunoassay for determining human interleukin-6 (IL-6) in biological fluids. The detection limit of the assay is 20 ng/L (B0 - 2 SD). Bound radioactivity in the range of 30% to 90% of the B0 counts corresponds to IL-6 concentrations of 100 to 14,000 ng/L. Analytical recovery of IL-6 added to EDTA-treated plasma averaged 25% more than that added to serum. The plasma concentration of IL-6 was therefore approximately 85 ng/L more than the concentration in simultaneously drawn serum. The mean serum concentration of IL-6 in 45 healthy subjects was 83 ng/L (range 20-290 ng/L), in 20 patients with multiple myeloma 303 ng/L, in 20 patients with rheumatoid arthritis 234 ng/L, and in 13 patients with systemic lupus erythematosus 183 ng/L. Markedly increased (greater than 3000 ng/L) concentrations of IL-6 were found in sera of patients with meningococcus meningitis and infectious peritonitis.


Empowering Site-Specific Bioconjugations In Vitro and In Vivo: Advances in Sortase Engineering and Sortase-Mediated Ligation.
Sortase-mediated ligation (SML) has emerged as a powerful and versatile methodology for site-specific protein conjugation, functionalization/labeling, immobilization, and design of biohybrid molecules and systems. However, the broader application of SML faces several challenges, such as limited activity and stability, dependence on calcium ions, and reversible reactions caused by nucleophilic side-products. Over the past decade, protein engineering campaigns and particularly directed evolution, have been extensively employed to overcome sortase limitations, thereby expanding the potential application of SML in multiple directions, including therapeutics, biorthogonal chemistry, biomaterials, and biosensors. This review provides an overview of achieved advancements in sortase engineering and highlights recent progress in utilizing SML in combination with other state-of-the-art chemical and biological methodologies. The aim is to encourage scientists to employ sortases in their conjugation experiments.


Maternal obesity in pregnancy and respiratory health in early childhood.
Obesity is associated with systemic inflammation, immunological changes, increased risk of respiratory infections and chronic respiratory illness. Maternal obesity in pregnancy increases the risk of pregnancy complications, caesarean sections and adverse birth outcomes, which have in turn been associated with respiratory illness in children. To our knowledge, the possible influence of maternal obesity in pregnancy on respiratory illness in early childhood beyond the newborn period has not been explored. We examined the relationship between a high maternal body mass index (BMI) in pregnancy and lower respiratory tract infections and wheeze up to 18 months of age in the Norwegian Mother and Child Study (MoBa), a population-based cohort study that includes 100,000 pregnant women, conducted at the Norwegian Institute of Public Health. We analysed data from the first 33 192 children, born between 1999 and 2005. In unadjusted analyses maternal obesity in pregnancy was related to both respiratory infections and wheeze in the children. In multivariable analyses, only an effect on wheeze remained. The risk of wheeze increased linearly with maternal BMI in pregnancy, and was 3.3% higher [95% CI 1.2, 5.3] for children with mothers who were obese during pregnancy, than for children of mothers with normal BMI. This effect was not mediated through obesity-related pregnancy complications, low birthweight, preterm birth or caesarean section.


Natural history and significance of esophageal squamous cell dysplasia.
Balloon-mesh cytologic screening for esophageal cancer done in 255 asymptomatic high-risk United States veterans (age greater than 40 years, ethanol abuse for greater than 20 years, and cigarette smoking greater than 20 pack years) identified 37 patients with squamous cell dysplasia. Of the 37 patients with dysplasia, 28 were re-evaluated prospectively at 6-month intervals for up to 36 months by balloon-mesh cytology, esophagoscopy with vital staining and biopsies, chest radiographs, oropharyngeal examination, and indirect laryngoscopy. During prospective follow-up evaluation, cytology specimens were repetitively normal in 16 patients (57%), showed inflammatory changes in eight patients (29%), persisted as dysplasia in two patients (7%) (both had endoscopic and histologic evidence of esophagitis), and progressed to carcinoma in two patients (7%) (one esophageal, one laryngeal). Although histologic findings concurred with the resolution of dysplasia, biopsy specimens were characterized by a similar difficulty in distinguishing dysplasia from inflammation. Erroneous histologic diagnoses of carcinoma in situ were made in two patients with reflux esophagitis evident endoscopically and confirmed during the course of a 24-36 month follow-up period. The authors conclude that squamous cell dysplasia detected by balloon-mesh cytology is seldom a precursor of esophageal cancer in the high-risk U.S. population but, rather, is often related to esophagitis. Thus, balloon-mesh cytology has limited use as a screening method for the early detection of esophageal cancer in the United States.


Flagellar phenotypic plasticity in volvocalean algae correlates with Péclet number.
Flagella-generated fluid stirring has been suggested to enhance nutrient uptake for sufficiently large micro-organisms, and to have played a role in evolutionary transitions to multicellularity. A corollary to this predicted size-dependent benefit is a propensity for phenotypic plasticity in the flow-generating mechanism to appear in large species under nutrient deprivation. We examined four species of volvocalean algae whose radii and flow speeds differ greatly, with Péclet numbers (Pe) separated by several orders of magnitude. Populations of unicellular Chlamydomonas reinhardtii and one- to eight-celled Gonium pectorale (Pe ∼ 0.1-1) and multicellular Volvox carteri and Volvox barberi (Pe ∼ 100) were grown in diluted and undiluted media. For C. reinhardtii and G. pectorale, decreasing the nutrient concentration resulted in smaller cells, but had no effect on flagellar length and propulsion force. In contrast, these conditions induced Volvox colonies to grow larger and increase their flagellar length, separating the somatic cells further. Detailed studies on V. carteri found that the opposing effects of increasing beating force and flagellar spacing balance, so the fluid speed across the colony surface remains unchanged between nutrient conditions. These results lend further support to the hypothesized link between the Péclet number, nutrient uptake and the evolution of biological complexity in the Volvocales.


Anomalous aortic origin of the pulmonary arteries: Case series and literature review.
Anomalous origin of the pulmonary arteries from the ascending aorta is a rare, but severe clinical entity necessitating a scrupulous evaluation. Either the right or the left pulmonary arteries can arise directly from the ascending aorta while the other pulmonary artery retains its origin from the right ventricular outflow tract. Such a finding can be isolated or can coexist with several congenital heart lesions. Direct intrapericardial aortic origin, however, must be distinguished with origin through a persistently patent arterial duct. In the current era, clinical manifestations usually become evident in the newborn rather than during infancy, as used to be the case. They include respiratory distress or congestive heart failure due to increased pulmonary flow and poor feeding. The rate of survival has now increased due to early diagnosis and prompt surgical repair, should now be expected to be at least 95%. We have treated four neonates with this lesion over the past 7 years, all of whom survived surgical repair. Right ventricular systolic pressure was significantly decreased at follow-up. Our choice of treatment was to translocate the anomalous pulmonary artery in end-to-side fashion to the pulmonary trunk. Our aim in this report is to update an Italian experience in the diagnosis and treatment of anomalous direct origin of one pulmonary artery from the aorta, adding considerations on the lessons learned from our most recent review of the salient literature.


Cell kinetic studies in mixed leukocyte cultures: an in vitro model of homograft reactivity.
The recognition phase of homograft immunity can be studied in vitro in the mixed leukocyte culture reaction. In this reaction, at the end of seven days, up to 30 per cent of the lymphocytes in culture "respond" to a single allogeneic cell stimulus. This paper presents evidence that the lymphocytes responding in culture divide with the generation time of 18-21 hours, with some asynchrony, and that a large percentage of the cells found at the end of the culture period may be the products by division of a small number of cells initially responding. Two estimates are made of the frequency of the initially responding unit (the cell(s) responsible for initiating the response). These are probably minimum estimates and are in the range of 1 in 200 to 1 in 2000 cells which can initially respond. This very high frequency of responding units as compared with the responding units in immediate type hypersensitivity (antibody production) is discussed.


Ingested fat oxidation contributes 8% of 24-h total energy expenditure in moderately obese subjects.
The role of ingested fat in the etiology of obesity is controversial. The aims of this study were to determine the contributions of ingested fat oxidation to: 1) 24-h total energy expenditure (TEE), and 2) substrate oxidation during acute stationary cycle exercises in adult humans. Healthy, moderately obese (n = 18; BMI = 31 +/- 1 kg/m2) subjects (8 men; 10 women) were each studied in a whole-room calorimeter for 24 h. They were fed mixed meals (55, 30, and 15% as energy from carbohydrate, fat and protein, respectively) to maintain energy balance. Each subject performed 1255-kJ cycle exercises at 50% VO2max in the calorimeter. Study test meal fat was labeled with carbon-13 (13C). Ingested fat oxidation was estimated from breath 13CO2 excretion and the subject's chamber CO2 production. Total fat and carbohydrate oxidations were estimated from nonprotein respiratory quotient (NP-RQ) values. Endogenous fat oxidation was estimated as the difference between total fat and ingested fat oxidations. TEE was estimated from gas exchanges; 28 +/- 3% of ingested fat was oxidized and it provided 8 +/- 1% of 24-h TEE. During cycle exercises, ingested fat provided 50% of total fat oxidized and 13.0 +/- 2% of energy expended. Endogenous fat oxidation contributed 10.4 +/- 3% of energy expenditure during cycle exercises. This study extended to 24-h observations of previous studies that lasted 6-9 h on ingested fat oxidation in humans. Understanding the factors that promote ingested fat oxidation could lead to more effective obesity intervention programs.


Effects of structural modifications of anti-tumour antibiotics luzopeptins on cell growth and macromolecule biosynthesis.
Luzopeptin analogues, A, B and C (in decreasing degree of acetylation) were active, less active and inactive, respectively, against several animal tumour models. All bound to isolated DNA avidly. The present studies with cultured Novikoff hepatoma cells showed that luzopeptins A, B and C were inhibitory, less inhibitory and ineffective, respectively, on cell colony formation ability and whole-cell DNA and RNA synthesis. RNA synthesis was more sensitive than DNA synthesis to luzopeptins A and B. All had no direct effect on protein synthesis. All inhibited the DNA and RNA syntheses of isolated nuclei and luzopeptins B and C were slightly more active than luzopeptin A inhibition of RNA synthesis in isolated nuclei. All similarly inhibited DNA polymerase activity in vitro, but luzopeptin C was more active against RNA polymerase activity in vitro. It is concluded that luzopeptin cytotoxicity probably resulted from inhibition of DNA and RNA biosynthesis. Because all analogues bound avidly to isolated DNA and inhibited its functions, the differential anti-tumour activity may be attributed to the differential ability of these luzopeptins to traverse the cell membrane and, to some extent, other intracellular barriers. This probably is a result of difference in acetylation, resulting in differences in hydrophobicity.


Dendritic cell vaccine with Ag85A enhances anti-colorectal carcinoma immunity.
Dendritic cells (DCs) are able to trigger T-cell activation and thus have been considered important for vaccine production against cancers. Vaccines containing DCs have been reported to be effective for developing immunity against cancer cells. The interactions between DCs and auxiliary agents are critical in the development of second-generation vaccines. In the present study, it was evaluated whether Ag85A-mixed DCs could enhance anti-tumor immunity in laboratory mice with colorectal carcinoma. Functional and phenotypic analyses of the effects of Ag85A-mixed DCs were conducted via flow cytometry and measurement of T-cell proliferation. In addition, interferon (IFN)-γ production was assessed. The therapeutic efficacy of DC vaccination for colorectal carcinoma treatment in mice was investigated. It was identified that Ag85A-mixed DCs exhibited strong upregulation of CD80, CD86 and major histocompatibility complex class II. Cytotoxic T-lymphocytes with CT26-primed Ag85A-DCs were indicated to induce stronger responses against CT26 tumor cells and trigger IFN-γ production. Furthermore, the Ag85A-mixed DC vaccine exerted a considerable inhibitory effect on tumor progression in mice as compared with the control group. Therefore, DCs in combination with the Ag85A gene may reinforce anti-colorectal carcinoma immunity. The current study provides a novel potential strategy for cancer treatment by enhancing immunity via Ag85A-mixed DC vaccination.


Sodium alginate microspheres of metformin HCl: formulation and in vitro evaluation.
Metformin microspheres with sodium alginate alone and in combination with gellan were prepared using an emulsion-cross linking method. The prepared microspheres were evaluated for their physico-chemical characteristics like particle size, morphology using SEM, incorporation efficiency, equilibrium water content (swelling) and in vitro drug release. The effect of various formulation variables like polymer concentration (sodium alginate; and proportion of gellan in microspheres prepared by a combination of sodium alginate and gellan), drug loading, crosslinking agent concentration and cross-linking time on the in vitro dissolution of the prepared microspheres were evaluated. The results showed that both the particle size and the incorporation efficiency were proportional to the polymer concentration. In case of microspheres containing both sodium alginate and gellan, the mean diameter and the incorporation efficiency were higher than the corresponding microspheres containing only alginate, both increasing with an increase in proportion of gellan. The prepared microspheres were found to be discrete and spherical in shape and were successful in sustaining the drug release for 8 hours. Incorporation of gellan caused a significant decrease in drug release. The release followed a biphasic profile, in all cases, characterized by an initial phase of moderate drug release followed by a phase of higher release. Further, the kinetic treatment of the dissolution data revealed the prevalence of matrix diffusion kinetics.


Psoriasis during therapy with olanzapine.
The role of several drugs (lithium, betablockers, clonidine, digoxin, nonsteroidal anti-inflammatory agents, interferon and tetracyclines) in inducing or exacerbating psoriasis is not completely understood. We report two cases of psoriasis in patients treated with olanzapine, a new drug used for psychotic disorders. In the first patient worsening of a pre-existing mild psoriasis occurred shortly after starting olanzapine treatment, followed by a relapse of the dermatological disease after a new administration of the anti-psychotic drug. In the second case we observed the onset of psoriasis during olanzapine treatment in an individual without any previous history of psoriasis but with a family history of the disease. Although the mechanism by which psoriasis may be induced or exacerbated by olanzapine remains unknown, these observations underline the need for caution and close follow-up in the pharmacological management of patients with psoriasis or a family history of the disease.


The upper thermal tolerance of the secondary screwworm, Cochliomyia macellaria Fabricius (Diptera: Calliphoridae).
Determining the thermal tolerance of an organism is important when assessing its activity time and survival rate in a given environment. However, thermal tolerance is not a static trait and may be influenced by a number of environmental and organismal factors. We report the results of three experiments investigating the effects of environmental temperature, exposure duration, age, sex, and nutrient availability on the upper thermal tolerance of the adult secondary screwworm, Cochliomyia macellaria. The probability of knockdown and survival was determined using a static method for different environmental temperatures (22, 40, 42, 44, or 45 °C), exposure durations (1, 2, 4, or 6 h), and nutrient availabilities (no food or water, water only, or both food and water) for both sexes and two age classes (young = 7-9 days post pupal emergence, old = 10-12 days post pupal emergence). In general, environmental temperature and exposure duration had the greatest effects on both the probability of knockdown and survival. As temperature or duration increased, the probability of knockdown increased while the probability of survival decreased. The availability of nutrients (water only or food and water) increased thermal tolerance at moderate temperatures (42 and 44 °C), but had no effect at 45 °C. Female flies were more thermally tolerant than males, regardless of nutrient availability. And age exhibited negligible effects on the probabilities of knockdown or survival, regardless of nutrient availability. These data show multiple environmental factors affected the thermal tolerance of C. macellaria. Thus, such aspects of basic thermal biology should feature more prominently in applied fields using blow flies, including but not limited to forensic entomology, disease ecology, and pollination ecology.


[Interdisciplinary recommendations on targeted therapy in the treatment of renal cell carcinoma].
Recently, new data have been published on the treatment of metastasized renal cell cancer using targeted therapies. With the approval of the tyrosine kinase inhibitors Sunitinib and Sorafenib, two of these new therapies are now available in clinical practice. This has raised both new opportunities and new questions for the health care professionals involved. Here we report on a consensus conference addressing these questions with answers based on evidence from the recent literature.


The pH of the host niche controls gene expression in and virulence of Candida albicans.
Little is known of the biological attributes conferring pathogenicity on the opportunistic fungal pathogen Candida albicans. Infection by this pathogen, as for bacterial pathogens, may rely upon environmental signals within the host niche to regulate the expression of virulence determinants. To determine if C. albicans responds to the pH of the host niche, we tested the virulence of strains with mutations in either of two pH-regulated genes, PHR1 and PHR2. In vitro, PHR1 is expressed when the ambient pH is at 5.5 or higher and deletion of the gene results in growth and morphological defects at neutral to alkaline pHs. Conversely, PHR2 is expressed at an ambient pH below 5.5, and the growth and morphology of the null mutant is compromised below this pH. A PHR1 null mutant was avirulent in a mouse model of systemic infection but uncompromised in its ability to cause vaginal infection in rats. Since systemic pH is near neutrality and vaginal pH is around 4.5, the virulence phenotype paralleled the pH dependence of the in vitro phenotypes. The virulence phenotype of a PHR2 null mutant was the inverse. The mutant was virulent in a systemic-infection model but avirulent in a vaginal-infection model. Heterozygous mutants exhibited partial reductions in their pathogenic potential, suggesting a gene dosage effect. Unexpectedly, deletion of PHR2 did not prevent hyphal development in vaginal tissue, suggesting that it is not essential for hyphal development in this host niche. The results suggest that the pH of the infection site regulates the expression of genes essential to survival within that niche. This implies that the study of environmentally regulated genes may provide a rationale for understanding the pathobiology of C. albicans.


Odontogenic tumours in children and adolescents: a review of forty-eight cases.
Odontogenic tumours comprise a large heterogeneous group of lesions originating from odontogenic epithelium and/or ectomesenchyme and its vestiges. The aim of this study was to analyze odontogenic tumours in children and adolescents seen at a tertiary institution in South Western Nigeria and compare with results from previous studies. Archival records of the Department of Oral Pathology, University College Hospital Ibadan were reviewed. All histologically diagnosed odontogenic tumours in patients 19 years and below spanning a period of 21 years (1990-2011) were retrieved. Data regarding age, gender, and tumor topography were analyzed using SPSS for Window (version 18.0; SPSS Inc. Chicago, IL). One hundred and forty seven jaw swellings were seen in children and adolescents aged 19 or less during the study period, out of which 48 (32.7%) were odontogenic tumours. More cases were seen in males than females with a male: female ratio of 7:5. The mandible was the commonest site of occurrence with mandible: maxilla ratio of 11:4. Ameloblastoma was the commonest odontogenic tumours with 14 (29.1%) solid ameloblastoma and 9 (18.8%) cystic ameloblastoma cases followed by fibromyxoma with 8 (16.7%) cases. calcifying epithelial odontogenic tumour , calcifying cystic odontogenic tumour and odontogenic fibroma were occasionally seen. This study showed that ameloblastoma was the most common odontogenic tumour in children and no case of odontoma was seen. Odontogenic tumours in children and adolescents may not be as rare as previously reported by some authors and inclusion of keratocystic odontogenic tumour in this study slightly affected the relative incidence of odontogenic tumors in children and adolescents.


Maternal-child consumption of ultra-processed foods and sugar-sweetened beverages in informal settlements in Mumbai, India.
The global nutrition transition is associated with increased consumption of ultra-processed snack foods and sugar-sweetened beverages (UPF/SSB), contributing to the double burden of child obesity and undernutrition. This cross-sectional study describes the prevalence of maternal and child UPF/SSB consumption and the factors associated with frequent consumption in a convenience sample of 749 children ages 6 months through 6 years and their mothers participating in a community-based child oral health program in five informal settlement communities in Mumbai, India. Mothers were interviewed regarding maternal and child oral health and nutrition characteristics, including consumption of beverages and foods associated with tooth decay-milk, soda, tea with sugar, sweets, and chips/biscuits-using standardized questionnaires. Spearman correlations were used to assess for associations between various social factors and the frequency of maternal and child consumption of the five food categories. Chi-square tests were used to assess differences in child consumption patterns by age groups. Though reported soda consumption was low among both mothers and children, nearly 60% of children consumed sweets and chips/biscuits daily, four to five times the rate of mothers. Factors associated with children's frequent consumption of UPF/SSB included lower maternal education level, frequent maternal consumption of UPF/SSB, greater number of household members, greater amount of money given to the child, and closer proximity to a store. Our findings demonstrate social factors that may promote UPF/SSB consumption. The nutritional dangers of sugary drinks and non-nutritious snacks for mothers and young children should be addressed across maternal-child health, education, and social service programs. Early childhood nutrition interventions should involve the entire family and community and emphasize the need to limit children's consumption of unhealthy foods and beverages from an early age.


Identification of essential glutamates in the acetate kinase from Methanosarcina thermophila.
Acetate kinase catalyzes the reversible phosphorylation of acetate (CH3COO- + ATP<-->CH3CO2PO3(2-) + ADP). A mechanism which involves a covalent phosphoryl-enzyme intermediate has been proposed, and chemical modification studies of the enzyme from Escherichia coli indicate an unspecified glutamate residue is phosphorylated (J. A. Todhunter and D. L. Purich, Biochem. Biophys. Res. Commun. 60:273-280, 1974). Alignment of the amino acid sequences for the acetate kinases from E. coli (Bacteria domain), Methanosarcina thermophila (Archaea domain), and four other phylogenetically divergent microbes revealed high identity which included five glutamates. These glutamates were replaced in the M. thermophila enzyme to determine if any are essential for catalysis. The histidine-tagged altered enzymes were produced in E. coli and purified to electrophoretic homogeneity by metal affinity chromatography. Replacements of E384 resulted in either undetectable or extremely low kinase activity, suggesting E384 is essential for catalysis which supports the proposed mechanism. Replacement of E385 influenced the Km values for acetate and ATP with only moderate decreases in k(cat), which suggests that this residue is involved in substrate binding but not catalysis. The unaltered acetate kinase was not inactivated by N-ethylmaleimide; however, replacement of E385 with cysteine conferred sensitivity to N-ethylmaleimide which was prevented by preincubation with acetate, acetyl phosphate, ATP, or ADP, suggesting that E385 is located near the active site. Replacement of E97 decreased the Km value for acetate but not ATP, suggesting this residue is involved in binding acetate. Replacement of either E32 or E334 had no significant effects on the kinetic constants, which indicates that neither residue is essential for catalysis or significantly influences the binding of acetate or ATP.


Environmental stress increases the invasion success of antimicrobial resistant bacteria in river microbial communities.
Environmental microbiomes are constantly exposed to invasion events through foreign, antibiotic resistant bacteria that were enriched in the anthropic sphere. However, the biotic and abiotic factors, as well as the natural barriers that determine the invasion success of these invader bacteria into the environmental microbiomes are poorly understood. A great example of such invasion events are river microbial communities constantly exposed to resistant bacteria originating from wastewater effluents. Here, we aim at gaining comprehensive insights into the key factors that determine their invasion success with a particular focus on the effects of environmental stressors, regularly co-released in wastewater effluents. Understanding invasion dynamics of resistant bacteria is crucial for limiting the environmental spread of antibiotic resistance. To achieve this, we grew natural microbial biofilms on glass slides in rivers for one month. The biofilms were then transferred to laboratory, recirculating flume systems and exposed to a single pulse of a model resistant invader bacterium (Escherichia coli) either in presence or absence of stress induced by Cu2+. The invasion dynamics of E. coli into the biofilms were then monitored for 14 days. Despite an initially successful introduction of E. coli into the biofilms, independent of the imposed stress, over time the invader perished in absence of stress. However, under stress the invading strain successfully established and proliferated in the biofilms. Noteworthy, the increased establishment success of the invader coincided with a loss in microbial community diversity under stress conditions, likely due to additional niche space becoming available for the invader.


A quantitative study of the effects of several nucleoside analogues on established herpes encephalitis in mice.
Mice with established herpes encephalitis were used to compare the effects of chemotherapy using three different nucleoside analogues. Encephalitis was produced by intranasal inoculation of a type 1 strain of herpes simplex virus. Without chemotherapy all mice died within 5 to 7 days of inoculation. Oral acyclovir (ACV) was a successful preventative measure if commenced within 2 days of inoculation but much less effective if the onset of treatment was further delayed. From the third day, when central nervous system infection had definitely become established, ACV only reduced mortality if given intraperitoneally (i.p.) at regular 6-hourly intervals. Comparison with bromovinyldeoxyuridine (BVdU) and the new nucleoside analogue dihydroxypropoxymethylguanine (DHPG) using the same 6-hourly i.p. regimen revealed that BVdU was poorly effective, despite better activity in vitro, whereas DHPG was the most successful. Virus was rapidly eradicated from all parts of the brain by DHPG therapy, and by day 10, no infectious virus remained in the brains of treated mice, no virus antigens were observed and no trace of virus DNA could be detected in neural tissues by Southern blotting.


Functional multispectral optoacoustic tomography imaging of hepatic steatosis development in mice.
The increasing worldwide prevalence of obesity, fatty liver diseases and the emerging understanding of the important roles lipids play in various other diseases is generating significant interest in lipid research. Lipid visualization in particular can play a critical role in understanding functional relations in lipid metabolism. We investigated the potential of multispectral optoacoustic tomography (MSOT) as a novel modality to non-invasively visualize lipids in laboratory mice around the 930nm spectral range. Using an obesity-induced non-alcoholic fatty liver disease (NAFLD) mouse model, we examined whether MSOT could detect and differentiate different grades of hepatic steatosis and monitor the accumulation of lipids in the liver quantitatively over time, without the use of contrast agents, i.e. in label-free mode. Moreover, we demonstrate the efficacy of using the real-time clearance kinetics of indocyanine green (ICG) in the liver, monitored by MSOT, as a biomarker to evaluate the organ's function and assess the severity of NAFLD. This study establishes MSOT as an efficient imaging tool for lipid visualization in preclinical studies, particularly for the assessment of NAFLD.


Structure and expression of the egg-laying hormone gene family in Aplysia.
Transcription of the egg-laying hormone (ELH) gene family was examined by characterizing homologous cDNA clones from abdominal ganglion and atrial gland cDNA libraries. All cDNAs contain an exon that spans the coding region (exon III) and one or two additional exons. The tissue-specific expression of the ELH gene family was confirmed by the observation that exon III encodes the ELH precursor protein in the bag cell transcripts and either the A or B precursor proteins in the atrial gland transcripts. The cDNA clones also contain 5' untranslated exons not present in the previously isolated genomic clones. One type of transcript has a 40-base pair segment, designated exon I, contiguous with exon III. A second type of transcript has an additional 149 base pairs of DNA, designated exon II, located between exons I and III. Several genomic clones containing exons I and II were isolated. DNA sequence analysis reveals that exons I and II are directly linked and that they are separated from exon III by an intervening sequence of at least 5 kilobases (kb). Consensus sequences for a putative promotor region and also for RNA splicing and polyadenylation were identified. From this work we can describe a prototype ELH gene complete with identified sequences necessary for the proper initiation of transcription and the subsequent processing of the transcript.


Holey Substrate-Directed Strain Patterning in Bilayer MoS2.
Key properties of two-dimensional (2D) layered materials are highly strain tunable, arising from bond modulation and associated reconfiguration of the energy bands around the Fermi level. Approaches to locally controlling and patterning strain have included both active and passive elastic deformation via sustained loading and templating with nanostructures. Here, by float-capturing ultrathin flakes of single-crystal 2H-MoS2 on amorphous holey silicon nitride substrates, we find that highly symmetric, high-fidelity strain patterns are formed. The hexagonally arranged holes and surface topography combine to generate highly conformal flake-substrate coverage creating patterns that match optimal centroidal Voronoi tessellation in 2D Euclidean space. Using TEM imaging and diffraction, as well as AFM topographic mapping, we determine that the substrate-driven 3D geometry of the flakes over the holes consists of symmetric, out-of-plane bowl-like deformation of up to 35 nm, with in-plane, isotropic tensile strains of up to 1.8% (measured with both selected-area diffraction and AFM). Atomistic and image simulations accurately predict spontaneous formation of the strain patterns, with van der Waals forces and substrate topography as the input parameters. These results show that predictable patterns and 3D topography can be spontaneously induced in 2D materials captured on bare, holey substrates. The method also enables electron scattering studies of precisely aligned, substrate-free strained regions in transmission mode.


Barriers and facilitators to physical activity among ethnic Chinese children: a qualitative systematic review.
The review aimed to synthesize the barriers and facilitators from the available studies that explored physical activity among ethnic Chinese children and uncover any differences or similarities in these barriers and facilitators. Physical activity promotes overall health, fitness, and well-being in children, yet prevalence of this has been low among ethnic Chinese children who reside in either Chinese and non-Chinese territories. Research has been conducted to explore the barriers and facilitators to physical activity among ethnic Chinese children. However, no qualitative systematic review has been conducted to synthesize these barriers and facilitators. Studies were considered for inclusion if they explored the barriers and facilitators to physical activity among ethnic Chinese children aged six to 17 years in either Chinese or non-Chinese territories, or among people who had responsibility for them in school, home, and community settings. The review included studies that focused on their views, experiences, attitudes, understandings, perceptions, and perspectives. Studies were included if they focused on qualitative data including, but not limited to, designs such as phenomenology, ethnography, grounded theory and action research. In addition, the authors considered cross-sectional surveys to find any free text relating to the review question. MEDLINE, Embase, CINAHL, PsycINFO, BNI, AMED, Web of Science, Scopus, CNKI, Wanfang and VIP databases were searched to identify published studies. The search for unpublished studies included EThOS, OpenGrey, ProQuest Dissertations and Theses, CNKI and Wanfang. Databases were searched from their inception dates to 10 December 2018 and no language restrictions were applied. The JBI guidelines for qualitative systematic reviews were followed in conducting the review. The JBI process of meta-aggregation was used to identify categories and synthesize findings. Out of 9460 records identified, 11 qualitative studies met the eligibility criteria and were included in the review. Using the JBI checklist for qualitative research (10 criteria), the critical appraisal scores of the majority of studies ranged from a moderate score of 6 (n = 1) to a high score of 7 and above (n = 9). Seven studies were from China, two from Australia, one each from the United Kingdom and the United States. The sample size ranged from 12 to 115 participants. A total of 56 findings were extracted and aggregated into 21 categories, based on the similarity of meaning. From studies conducted in the Chinese territories, four synthesized findings (personal, socio-cultural, environmental, and policy- and program-related barriers and facilitators) were aggregated from 37 extracted findings and 14 aggregated categories. From studies conducted in the non-Chinese territories, only two synthesized findings (personal and socio-cultural barriers and facilitators) were derived from 19 extracted findings and seven aggregated categories. Based on the ConQual scores, confidence in the synthesized findings was moderate. Four broad themes emerged from the participants' accounts, namely personal, socio-cultural, environmental, and policy- and program-related factors. Barriers and facilitators at the personal and socio-cultural level (e.g., parents and teachers) were most frequently cited, reflecting the importance of children's self-influence and the role of adults. Future interventions are needed to address the identified barriers and enhance the facilitators. PROSPERO CRD42018097124.


(2E)-1-(2,4,6-Trimethoxyphenyl)-3-(4-chlorophenyl)prop-2-en-1-one, a Chalcone Derivative, Promotes Apoptosis by Suppressing RAS-ERK and AKT/FOXO3a Pathways in Hepatocellular Carcinoma Cells.
Liver cancer is an extremely common cancer with the highest mortality rate and poor prognosis. Owing to their low systemic toxicity and few side effects, natural compounds may provide better therapeutic effects for patients. (2E)-1-(2,4,6-trimethoxyphenyl)-3-(4-chlorophenyl)prop-2-en-1-one (TMOCC), a chalcone derivative, exhibits cytotoxicity towards many tumor cells. However, the anticancer mechanism of TMOCC has not been elucidated in human hepatocellular carcinoma (HCC). Cell Counting Kit-8 and colony formation assays were used to evaluate the effects of TMOCC on viability and proliferation. Mitochondrial transmembrane potential and flow cytometry assays were used to detect apoptosis. The expression levels of proteins related to apoptosis, the RAS-ERK and AKT/FOXO3a signaling pathways were assessed using western blot. Potential targets of TMOCC were detected using molecular docking analysis. TMOCC inhibited viability and proliferation, and induced the loss of mitochondrial transmembrane potential, apoptosis and DNA double-strand breaks in both HCC cells. The RAS-ERK and AKT/FOXO3a signaling pathways were suppressed by TMOCC. Finally, ERK1, PARP-1, and BAX were identified as potential targets of TMOCC. Taken together, our results show that TMOCC promotes apoptosis by suppressing the RAS-ERK and AKT/FOXO3a signaling pathways. TMOCC may be a potential multi-target compound that is effective against liver cancer.


Chemical Structure of Fe-Ni Nanoparticles for Efficient Oxygen Evolution Reaction Electrocatalysis.
Bimetallic iron-nickel-based nanocatalysts are perhaps the most active for the oxygen evolution reaction (OER) in alkaline electrolytes. Recent developments in literature have suggested that the ratio of iron and nickel in Fe-Ni thin films plays an essential role in the performance and stability of the catalysts. In this work, the metallic ratio of iron to nickel was tested in alloy bimetallic nanoparticles. Similar to thin films, nanoparticles with iron-nickel atomic compositions where the atomic iron percentage is ≤50% outperformed nanoparticles with iron-nickel ratios of >50%. Nanoparticles of Fe20Ni80, Fe50Ni50, and Fe80Ni20 compositions were evaluated and demonstrated to have overpotentials of 313, 327,, and 364 mV, respectively, at a current density of 10 mA/cm2. While the Fe20Ni80 composition might be considered to have the best OER performance at low current densities, Fe50Ni50 was found to have the best current density performance at higher current densities, making this composition particularly relevant for electrolysis conditions. However, when stability was evaluated through chronoamperometry and chronopotentiometry, the Fe80Ni20 composition resulted in the lowest degradation rates of 2.9 μA/h and 17.2 μV/h, respectively. These results suggest that nanoparticles with higher iron and lower nickel content, such as the Fe80Ni20 composition, should be still taken into consideration while optimizing these bimetallic OER catalysts for overall electrocatalytic performance. Characterization by electron microscopy, diffraction, and X-ray spectroscopy provides detailed chemical and structural information on as-synthesized nanoparticle materials.


Blockade of CTLA-4 decreases the generation of multifunctional memory CD4+ T cells in vivo.
CTLA-4 is known as a central inhibitor of T cell responses. It terminates T cell activation and proliferation and induces resistance against activation induced cell death. However, its impact on memory formation of adaptive immune responses is still unknown. In this study, we demonstrate that although anti-CTLA-4 mAb treatment during primary immunization of mice initially enhances the number of IFN-γ-producing CD4(+) T cells, it does not affect the size of the memory pool. Interestingly, we find that the CTLA-4 blockade modulates the quality of the memory pool: it decreases the amount of specialized "multifunctional" memory CD4(+) T cells coproducing IFN-γ, TNF-α, and IL-2 in response to Ag. The reduction of these cells causes an immense decrease of IFN-γ-producing T cells after in vivo antigenic rechallenge. Chimeric mice expressing CTLA-4-competent and -deficient cells unmask, which these CTLA-4-driven mechanisms are mediated CD4(+) T cell nonautonomously. In addition, the depletion of CD25(+) T cells prior to the generation of Ag-specific memory cells reveals that the constitutively CTLA-4-expressing natural regulatory T cells determine the quality of memory CD4(+) T cells. Taken together, these results indicate that although the inhibitory molecule CTLA-4 damps the primary immune response, its engagement positively regulates the formation of a high-quality memory pool equipped with multifunctional CD4(+) T cells capable of mounting a robust response to Ag rechallenge.


Essential role for the p55 tumor necrosis factor receptor in regulating hematopoiesis at a stem cell level.
Hematopoietic stem cell (HSC) self-renewal is a complicated process, and its regulatory mechanisms are poorly understood. Previous studies have identified tumor necrosis factor (TNF)-alpha as a pleiotropic cytokine, which, among other actions, prevents various hematopoietic progenitor cells from proliferating and differentiating in vitro. However, its role in regulating long-term repopulating HSCs in vivo has not been investigated. In this study, mice deficient for the p55 or the p75 subunit of the TNF receptor were analyzed in a variety of hematopoietic progenitor and stem cell assays. In older p55(-/-) mice (>6 mo), we identified significant differences in their hematopoietic system compared with age-matched p75(-/-) or wild-type counterparts. Increased marrow cellularity and increased numbers of myeloid and erythroid colony-forming progenitor cells (CFCs), paralleled by elevated peripheral blood cell counts, were found in p55-deficient mice. In contrast to the increased myeloid compartment, pre-B CFCs were deficient in older p55(-/-) mice. In addition, a fourfold decrease in the number of HSCs could be demonstrated in a competitive repopulating assay. Secondary transplantations of marrow cells from primary recipients of p55(-/-) marrow revealed impaired self-renewal ability of p55-deficient HSCs. These data show that, in vivo, signaling through the p55 subunit of the TNF receptor is essential for regulating hematopoiesis at the stem cell level.


Recombinant human granulocyte colony-stimulating factor reduces colonic epithelial cell apoptosis and ameliorates murine dextran sulfate sodium-induced colitis.
Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is a potentially effective therapy for Crohn's disease. The purpose of this study was to test the rhG-CSF in murine dextran sulfate sodium-induced colitis (DSS colitis). Murine colitis was induced by feeding with water containing 3% DSS for 9 days. Six to 7-week-old female BALB/c mice were given rhG-CSF (100 microg/kg) or phosphate-buffered saline (PBS) subcutaneously once a day from day 0 to day 8, and the mice were sacrificed at days 3, 5, 7 and 9. Tissue specimens from the transverse colon, descending colon and rectum were obtained and stained with hematoxylin and eosin. Inflammation was scored for severity, extent, epithelial damage and crypt loss. TUNEL staining was performed to assess epithelial cell apoptosis. Treatment with rhG-CSF significantly attenuated body-weight loss, stool score and shortening of the colon length in comparison with treatment with PBS (p<0.01,<0.05,<0.01, respectively). Histological scores for inflammation, epithelial cell damage and crypt loss of the rectum were less severe at day 9 in the rhG-CSF group than in the PBS group (p<0.01, 0.05, 0.01, respectively). The number of TUNEL-positive cells in the rectum was smaller in the rhG-CSF group than in the PBS group (p<0.001). Treatment with rhG-CSF ameliorates murine DSS colitis by suppressing mucosal inflammation and epithelial damage in the rectum. The prevention of epithelial cell apoptosis seems to precede the anti-inflammatory action of rhG-CSF.


Tensile properties of bioactive fibers for tissue engineering applications.
Cell transplantation using biocompatible, biodegradable scaffolds offers the possibility of creating or regenerating tissue to replace organ function when deficiency arises. The role of these temporary substrates is to support and guide the expanding cell culture until it becomes structurally integrated with the host tissue. 45S5 Bioglass(R) is a 4-component, melt-derived bioactive glass, which has been approved for human clinical use by the Food and Drug Administration. The biocompatibility and biodegradability of 45S5 Bioglass(R) are long established, whereas research into its performance as an extracellular scaffold is currently underway. In this study the tensile strengths (93 +/- 8 and 82 +/- 14 MPa), elongation to fracture (0.7 +/- 0.05%) and Weibull's moduli (3.0 and 3.5) of 45S5 Bioglass(R) fibers (mean diameters 193 and 280 microm) for tissue engineering applications are reported. The tensile strengths of the fibers are compared with those of bulk 45S5 Bioglass(R) and a range of biodegradable polymer materials currently used in the field of tissue engineering. Aspects of glass and fiber technology relevant to the design and manufacture of extracellular ceramic scaffolds are also discussed.


Observer agreement analysis of MRI grading of knee osteoarthritis.
The aim of the study was to use Kappa statistical analysis to assess the inter and intraobserver agreement of the interpretation of the osteoarthritis changes observed on a FISP sequence, 1.0 Tesla, MRI analysis of the articular cartilage in 30 knees. The images were read on two separate occasions by three observers. The best agreement was seen in the patello-femoral compartment. The most experienced of the assessors produced the more consistent results. The results of the interobserver agreement had mainly "slight" or "fair" agreement. Our results are disappointing and accordingly, we have reservations about the use of MRI in the assessment of osteoarthritis of the knee.


Primary prevention of sudden cardiac death using implantable cardioverter defibrillators.
Despite substantial advances in prevention and treatment of cardiovascular diseases, sudden cardiac death (SCD) remains a leading cause of death in industrialized countries. Implantable cardioverter defibrillator (ICD) has been demonstrated to be an attractive option for primary prevention of SCD in high-risk patients. This review discusses the progress in the risk stratification for selecting high-risk patients, highlights the clinical trials of primary prevention for SCD, outlines the efficacy of combined use of cardiac resynchronization therapy with ICD, and analyses the cost-effectiveness issue of this device.


Fractional laser combined with 5-Aminolevulinic acid photodynamic therapy for the treatment of Cutaneous Rosai-Dorfman Disease: A case report.
Rosai-Dorfman disease(RDD) is a rare benign histiocytic proliferative disorder. Cutaneous Rosai-Dorfman disease(CRDD)is a rare variant of RDD with lesions localized to the skin. We report a case of patients with CRDD who was refractory to methotrexate, thalidomide, cyclosporine, glucocorticoid for local injection and external use, but resolved with fractional laser combined with 5-aminolevulinic acid photodynamic therapy (ALA-PDT). We believe that fractional laser technology combined with ALA-PDT for CRDD is a minimally invasive, effective and satisfactory treatment.


Depression, anxiety, and quality of life in patients with atrial fibrillation.
To examine the prevalence and persistence of depression and anxiety in patients with atrial fibrillation (AF), and their effect on future quality of life (QoL) status. The Beck Depression Inventory and State-Trait Anxiety Inventory were completed by 101 patients with AF (62 men; mean age +/- SD, 66.3 +/- 11.0 years), who were compared to 97 patients with hypertension (as "disease control" subjects) in sinus rhythm (64 men; mean age, 68.0 +/- 7.2 years) at baseline and at 6 months. QoL was ascertained at both time points using Dartmouth Care Cooperative Information Project charts. At baseline among AF patients, symptoms of depression, state anxiety, and trait anxiety prevailed in 38%, 28%, and 38%, respectively; analogous data for hypertensive patients were 30%, 23%, and 22%. AF patients displayed higher levels of trait anxiety (p < 0.05), with no significant differences in baseline depression, state anxiety, and QoL between patients with AF and disease control subjects. Symptoms of depression and anxiety (state and trait) persisted at 6 months in 36.8% and 33.3%, respectively. Symptoms of depression (p < 0.001) and anxiety (p < 0.001) at baseline, female gender (p = 0.01), ethnicity (p = 0.01), and employment status (p = 0.03) were significantly correlated with QoL at 6 months in the patients with AF. Multiple regression analysis revealed that baseline depression score provided the best independent prediction of 6-month QoL (R(2) = 0.20), although gender and employment status also entered the model. Approximately one third of AF patients have elevated levels of depression and anxiety, which persist at 6 months. Symptoms of depression were the strongest independent predictor of future QoL in these patients.


Free-Form Liquid Crystal Elastomers via Embedded 4D Printing.
Liquid crystal elastomers (LCEs) are a class of active materials that can generate rapid, reversible mechanical actuation in response to external stimuli. Fabrication methods for LCEs have remained a topic of intense research interest in recent years. One promising approach, termed 4D printing, combines the advantages of 3D printing with responsive materials, such as LCEs, to generate smart structures that not only possess user-defined static shapes but also can change their shape over time. To date, 4D-printed LCE structures have been limited to flat objects, restricting shape complexity and associated actuation for smart structure applications. In this work, we report the development of embedded 4D printing to extrude hydrophobic LCE ink into an aqueous, thixotropic gel matrix to produce free-standing, free-form 3D architectures without sacrificing the mechanical actuation properties. The ability to 4D print complex, free-standing 3D LCE architectures opens new avenues for the design and development of functional and responsive systems, such as reconfigurable metamaterials, soft robotics, or biomedical devices.


Brain-computer interface users speak up: the Virtual Users' Forum at the 2013 International Brain-Computer Interface Meeting.
More than 300 researchers gathered at the 2013 International Brain-Computer Interface (BCI) Meeting to discuss current practice and future goals for BCI research and development. The authors organized the Virtual Users' Forum at the meeting to provide the BCI community with feedback from users. We report on the Virtual Users' Forum, including initial results from ongoing research being conducted by 2 BCI groups. Online surveys and in-person interviews were used to solicit feedback from people with disabilities who are expert and novice BCI users. For the Virtual Users' Forum, their responses were organized into 4 major themes: current (non-BCI) communication methods, experiences with BCI research, challenges of current BCIs, and future BCI developments. Two authors with severe disabilities gave presentations during the Virtual Users' Forum, and their comments are integrated with the other results. While participants' hopes for BCIs of the future remain high, their comments about available systems mirror those made by consumers about conventional assistive technology. They reflect concerns about reliability (eg, typing accuracy/speed), utility (eg, applications and the desire for real-time interactions), ease of use (eg, portability and system setup), and support (eg, technical support and caregiver training). People with disabilities, as target users of BCI systems, can provide valuable feedback and input on the development of BCI as an assistive technology. To this end, participatory action research should be considered as a valuable methodology for future BCI research.


Association Between Endothelial Cell Stabilizing Medication and Small Vessel Disease Stroke: A Case-Control Study.
Increasing evidence suggests a role for endothelial cell (EC) dysfunction in pathogenesis of cerebral small vessel disease. Commonly used medications including certain antihypertensives and statins have EC-stabilizing effects. We used individual patient data from completed acute stroke trials to assess whether prior exposure to EC-stabilizing medications was associated with lacunar stroke, using lacunar stroke as a clinical proxy for cerebral small vessel disease. Across 12,002 patients with relevant data, 2,855 (24%) had a lacunar stroke presentation. Univariable analyses suggested potential confounding from vascular diseases treated with EC-stabilizing medications. Initial multivariable logistic regression gave conflicting results when describing the independent association of exposure to EC-stabilizing medication and lacunar stroke in the complete population (O.R. 0.87, 95% C.I.: 0.77- 0.98) and limited to those taking any antihypertensive (O.R. 1.51, 95% C.I.: 1.21-1.88). Re-running the analyses including statins in the EC-stabilizing category suggested a beneficial effect of EC-stabilizing medication exposure on lacunar stroke incidence (O.R. 0.83, 95% C.I.: 0.73-0.93). These results align with recent pre-clinical data and would support interventional trials of EC-stabilizing medication for preventing cerebral small vessel disease. Our results also suggest that analyses of EC-stabilizing interventions need to adjust for potential endothelial effects of other co-prescribed medication.


Regulation of myocardin factor protein stability by the LIM-only protein FHL2.
Extensive evidence indicates that serum response factor (SRF) regulates muscle-specific gene expression and that myocardin family SRF cofactors are critical for smooth muscle cell differentiation. In a yeast two hybrid screen for novel SRF binding partners expressed in aortic SMC, we identified four and a half LIM domain protein 2 (FHL2) and confirmed this interaction by GST pull-down and coimmunoprecipitation assays. FHL2 also interacted with all three myocardin factors and enhanced myocardin and myocardin-related transcription factor (MRTF)-A-dependent transactivation of smooth muscle alpha-actin, SM22, and cardiac atrial natriuretic factor promoters in 10T1/2 cells. The expression of FHL2 increased myocardin and MRTF-A protein levels, and, importantly, this effect was due to an increase in protein stability not due to an increase in myocardin factor mRNA expression. Treatment of cells with proteasome inhibitors MG-132 and lactacystin strongly upregulated endogenous MRTF-A protein levels and resulted in a substantial increase in ubiquitin immunoreactivity in MRTF-A immunoprecipitants. Interestingly, the expression of FHL2 attenuated the effects of RhoA and MRTF-B on promoter activity, perhaps through decreased MRTF-B nuclear localization or decreased SRF-CArG binding. Taken together, these data indicate that myocardin factors are regulated by proteasome-mediated degradation and that FHL2 regulates SRF-dependent transcription by multiple mechanisms, including stabilization of myocardin and MRTF-A.


Analysis of circulating immune complexes from patients with ankylosing spondylitis by gel electrophoresis and immunoblotting using antiserum against a psoriasis associated retrovirus-like particle.
Circulating immune complexes (CIC) were isolated from patients with ankylosing spondylitis (AS) and healthy blood donors by isopycnic ultracentrifugation in sucrose gradients. The CIC were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. The major components of the CIC were identified as albumin, immunoglobulins, and complement factors. A 70 kD component and several low molecular weight components (Mr 19 kD and 14 kD (doublet] were detectable only in CIC from patients with AS. An antiserum raised against the envelope glycoprotein, gp70, of a psoriasis associated retrovirus-like particle was applied to check for cross reacting activity. This antiserum reacted with both a 70 kD and a 40-45 kD component in CIC from three out of six patients but not with CIC from any of the blood donors.


Localized modes in photonic quasicrystals with Penrose-type lattice.
We investigate the properties of the resonant modes that occur in the transparency bands of two-dimensional finite-size Penrose-type photonic quasicrystals made of dielectric cylindrical rods. These modes stem from the natural local arrangements of the quasicrystal structure rather than, as originally thought, from fabrication-related imperfections. Examples of local density of states and field maps are shown for different wavelengths. Calculations of local density of states show that these modes mainly originate from the interactions between a limited numbers of rods.


Keep and promote biodiversity at polluted sites under phytomanagement.
The phytomanagement concept combines a sustainable reduction of pollutant linkages at risk-assessed contaminated sites with the generation of both valuable biomass for the (bio)economy and ecosystem services. One of the potential benefits of phytomanagement is the possibility to increase biodiversity in polluted sites. However, the unique biodiversity present in some polluted sites can be severely impacted by the implementation of phytomanagement practices, even resulting in the local extinction of endemic ecotypes or species of great conservation value. Here, we highlight the importance of promoting measures to minimise the potential adverse impact of phytomanagement on biodiversity at polluted sites, as well as recommend practices to increase biodiversity at phytomanaged sites without compromising its effectiveness in terms of reduction of pollutant linkages and the generation of valuable biomass and ecosystem services.


Pathway Markers for Pro-resolving Lipid Mediators in Maternal and Umbilical Cord Blood: A Secondary Analysis of the Mothers, Omega-3, and Mental Health Study.
The omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) are precursors to immune regulatory and specialized pro-resolving mediators (SPM) of inflammation termed resolvins, maresins, and protectins. Evidence for lipid mediator formation in vivo can be gained through evaluation of their 5-lipoxygenase (LOX) and 15-LOX metabolic pathway precursors and downstream metabolites. We performed a secondary blood sample analysis from 60 participants in the Mothers, Omega-3, and Mental Health study to determine whether SPM and SPM precursors are augmented by dietary EPA- and DHA-rich fish oil supplementation compared to soy oil placebo. We also aimed to study whether SPM and their precursors differ in early and late pregnancy or between maternal and umbilical cord blood. We found that compared to placebo supplementation, EPA- and DHA-rich fish oil supplementation increased SPM precursor 17-hydroxy docosahexaenoic acid (17-HDHA) concentrations in maternal and umbilical cord blood (P = 0.02). We found that the D-series resolvin pathway marker 17-HDHA increased significantly between enrollment and late pregnancy (P = 0.049). Levels of both 14-HDHA, a maresin pathway marker, and 17-HDHA were significantly greater in umbilical cord blood than in maternal blood (P < 0.001, both).


Chemical characterization and bioactive properties of Geranium molle L.: from the plant to the most active extract and its phytochemicals.
After a period of indifference, in which synthetic compounds were favored, there is an increasing interest in the study of the biological properties of plants and the active principles responsible for their therapeutic properties. Geranium molle L. has been used in the Portuguese folk medicine for the treatment of various ailments including cancer but, unlike many of the species from the Geranium genus, its phytochemical characterization and biological activity are virtually unexplored. In this study a G. molle sample from Trás-os-Montes, north-eastern Portugal, was chemically characterized regarding nutritional value, free sugars, organic acids, fatty acids and tocopherols, and several aqueous (decoction, infusion) and organic (n-hexane, dichloromethane, ethyl acetate, acetone, methanol) extracts of the plant were assessed for their bioactive properties. The antioxidant activity was evaluated by means of the free radicals scavenging activity, reducing power and inhibition of lipid peroxidation. The cytotoxicity of the different extracts was assessed in vitro against several human cancer cell lines (breast, lung, cervical and hepatocellular carcinomas) and, additionally, their hepatotoxicity was evaluated using a porcine liver primary cell culture. G. molle was shown to be rich in carbohydrates and proteins, providing tocopherols and essential fatty acids. Amongst the various extracts, the acetone extract was found to have the highest content of phenolic compounds (mainly ellagitannins, but also some flavone and flavonol glycosides) as well as the highest antioxidant and cytotoxic activities. To the best of our knowledge, this is the first report on the chemical composition and bioactive properties of G. molle.


RSV-Induced H3K4 Demethylase KDM5B Leads to Regulation of Dendritic Cell-Derived Innate Cytokines and Exacerbates Pathogenesis In Vivo.
Respiratory syncytial virus (RSV) infection can result in severe disease partially due to its ability to interfere with the initiation of Th1 responses targeting the production of type I interferons (IFN) and promoting a Th2 immune environment. Epigenetic modulation of gene transcription has been shown to be important in regulating inflammatory pathways. RSV-infected bone marrow-derived DCs (BMDCs) upregulated expression of Kdm5b/Jarid1b H3K4 demethylase. Kdm5b-specific siRNA inhibition in BMDC led to a 10-fold increase in IFN-β as well as increases in IL-6 and TNF-α compared to control-transfected cells. The generation of Kdm5bfl/fl-CD11c-Cre+ mice recapitulated the latter results during in vitro DC activation showing innate cytokine modulation. In vivo, infection of Kdm5bfl/fl-CD11c-Cre+ mice with RSV resulted in higher production of IFN-γ and reduced IL-4 and IL-5 compared to littermate controls, with significantly decreased inflammation, IL-13, and mucus production in the lungs. Sensitization with RSV-infected DCs into the airways of naïve mice led to an exacerbated response when mice were challenged with live RSV infection. When Kdm5b was blocked in DCs with siRNA or DCs from Kdm5bfl/fl-CD11c-CRE mice were used, the exacerbated response was abrogated. Importantly, human monocyte-derived DCs treated with a chemical inhibitor for KDM5B resulted in increased innate cytokine levels as well as elicited decreased Th2 cytokines when co-cultured with RSV reactivated CD4+ T cells. These results suggest that KDM5B acts to repress type I IFN and other innate cytokines to promote an altered immune response following RSV infection that contributes to development of chronic disease.


UK population norms for the modified dental anxiety scale with percentile calculator: adult dental health survey 2009 results.
A recent UK population survey of oral health included questions to assess dental anxiety to provide mean and prevalence estimates of this important psychological construct. A two-stage cluster sample was used for the survey across England, Wales, and Northern Ireland. The survey took place between October-December 2009, and January-April 2010. All interviewers were trained on survey procedures. Within the 7,233 households sampled there were 13,509 adults who were asked to participate in the survey and 11,382 participated (84%). The scale was reliable and showed some evidence of unidimensionality. Estimated proportion of participants with high dental anxiety (cut-off score = 19) was 11.6%. Percentiles and confidence intervals were presented and can be estimated for individual patients across various age ranges and gender using an on-line tool. The largest reported data set on the MDAS from a representative UK sample was presented. The scale's psychometrics is supportive for the routine assessment of patient dental anxiety to compare against a number of major demographic groups categorised by age and sex. Practitioners within the UK have a resource to estimate the rarity of a particular patient's level of dental anxiety, with confidence intervals, when using the on-line percentile calculator.


Eradication of elephant ear mites (Loxoanoetus bassoni) in two African elephants (Loxodonta africana).
Elephant ear mites, not previously described in North America, were eradicated in two African elephants (Loxodonta africana) after six otic instillations of ivermectin at 2-wk intervals. The microscopic examination of a clear, mucoid discharge collected from the external ear canals of two wild-born African elephants housed in a New York State zoo for 25 yr revealed live mites (Loxoaneotus bassoni). The cytologic examination demonstrated no evidence of inflammation or infection. Both elephants were asymptomatic with normal hemograms and serum chemistry panels. A diagnosis of otoacariasis was made. Each elephant was treated six times with 5 ml of 1% ivermectin syrup instilled in each ear canal once every 2 wk. Microscopic examinations of clear mucus collected from each elephant's ear canals 9 days after the first instillation of ivermectin were negative for any life stages of ear mites. Microscopic examinations of mucus collected from both elephants' ear canals at 6, 11, and 16 wk, as well as annually post-treatment for 7 yr, confirmed eradication of the ear mites. The L. bassoni ear mite was first identified in the external ear canals of wild, asymptomatic, lesion-free, African elephants culled in Kruger National Park in South Africa. However, a new species in the same genus of mites (Loxoanoetus lenae) was identified at the necropsy of an 86-yr-old Asian elephant (Elephas maximus) living in a circus in Australia. The autopsy revealed a marked, ballooning distension of bone around the left external acoustic meatus, suggestive of mite-induced otitis externa, as seen in cattle infested with ear mites (Raillieta auris). Elephant health care providers should identify the prevalence of, and consider treatment of, elephants in their care infested with ear mites, given the possible risk for adverse health effects.


Accuracy of dipyridamole SPECT imaging in identifying individual coronary stenoses and multivessel disease in women versus men.
Older women frequently undergo dipyridamole perfusion imaging and can have advanced coronary artery disease, but little data exist on the accuracy of perfusion imaging in detecting disease in individual vascular territories and multivessel disease in women, compared with men. From a database of patients undergoing myocardial single photon emission computed tomography (SPECT) perfusion imaging, 107 unselected sequential patients (58 women, 49 men) who underwent sestamibi dipyridamole stress and cardiac catheterization within 6 months of each other were identified. Data were analyzed to compare sensitivities for detection of individual coronary stenoses and multivessel disease. The concordance between perfusion image results and cardiac catheterization for individual coronary territories for women was 75%, and for men, it was 65% (P = .09). In women, the presence of disease of the left anterior descending coronary artery was detected more frequently than it was in men, 84% versus 44% (P = .004). The detection of disease in the territories of the left circumflex and right coronary arteries was similar for both groups. For women, the accuracy of perfusion imaging in identifying the presence/absence of multivessel coronary disease was 64%, compared with 71 % for men (P = not significant). The accuracy of dipyridamole sestamibi SPECT imaging in detecting multivessel disease was similar for men and women. The sensitivity of dipyridamole sestamibi SPECT imaging in detecting disease of the left anterior descending artery was better in women.


U structured network with three encoding paths for breast tumor segmentation.
Breast ultrasound segmentation remains challenging because of the blurred boundaries, irregular shapes, and the presence of shadowing and speckle noise. The majority of approaches stack convolutional layers to extract advanced semantic information, which makes it difficult to handle multiscale issues. To address those issues, we propose a three-path U-structure network (TPUNet) that consists of a three-path encoder and an attention-based feature fusion block (AFF Block). Specifically, instead of simply stacking convolutional layers, we design a three-path encoder to capture multiscale features through three independent encoding paths. Additionally, we design an attention-based feature fusion block to weight and fuse feature maps in spatial and channel dimensions. The AFF Block encourages different paths to compete with each other in order to synthesize more salient feature maps. We also investigate a hybrid loss function for reducing false negative regions and refining the boundary segmentation, as well as the deep supervision to guide different paths to capture the effective features under the corresponding receptive field sizes. According to experimental findings, our proposed TPUNet achieves more excellent results in terms of quantitative analysis and visual quality than other rival approaches.


Cloning and sequencing of the light chain variable region from NS-1 myeloma.
The aim of this study was to clone the gene encoding the light chain variable region (V(L)) of the murine myeloma cell line P3/NS1/1-Ag4-1 (NS-1). Total RNA was prepared from the NS-1 cell line and its fusion hybridoma cell line 2F9. The V(L) gene was amplified using reverse transcription PCR (RT-PCR) with family-specific primer pairs. The PCR products were cloned into the pGEM(®)-T easy vector, transfected into E. coli DH5α cells and the positive recombinants were identified and purified with EcoRI digestion and agarose gel electrophoresis. The sequence was identified using an automatic DNA sequencer and analyzed online using the IMGT/V-QUEST program (version 3.2.21). The PCR product that was amplified with the P9 and P14 primers was approximately 392 bp in size. Following digestion with EcoRI, the band of interest was also detected in positive recombinants using agarose gel electrophoresis. The sequences from the NS-1 and 2F9 cells were identical. The whole sequence was 387 bp long, encoding 128 amino acids (AA), including a 60 bp leader sequence. There was a TAA stop codon at 385-387 bp and only one cysteine was found, at 112AA/128AA. Analysis using IMGT/V-QUEST determined the cloned V- and J-segments as murine IG(κ)KV3-12*01 and IG(κ)KJ2*01, respectively. Accordingly, the NS-1 V(L) gene belongs to the Ig(κ) gene family V3 subgroup. The NS-1 V(L) gene was successfully cloned and it is a pseudo-Igκ chain gene in NS-1 cells. This study may aid the sequencing of genes encoding monoclonal antibodies produced by mouse hybridomas raised with NS-1 myeloma.


Structural identification of mouse urinary metabolites of pterostilbene using liquid chromatography/tandem mass spectrometry.
Pterostilbene, the dimethoxy derivative of resveratrol, has drawn much attention recently due to its potential beneficial health effects. The metabolic fate of pterostilbene, however, is not well understood. In the present study, we identified nine novel mouse urinary pterostilbene metabolites, pterostilbene glucuronide, pterostilbene sulfate, mono-demethylated pterostilbene glucuronide, mono-demethylated pterostilbene sulfate, mono-hydroxylated pterostilbene, mono-hydroxylated pterostilbene glucuronide, mono-hydroxylated pterostilbene sulfate, and mono-hydroxylated pterostilbene glucuronide sulfate, using liquid chromatography/atmospheric pressure chemical ionization and electrospray ionization tandem mass spectrometry. The structures of these metabolites were confirmed by analyzing the MS(n) (n = 1-3) spectra. To our knowledge, this is the first report of the identification of urinary metabolites of pterostilbene in mice.


[Oral premedication with midasolam and ketamine in children with neurosurgical diseases].
The study was undertaken to evaluate the efficiency and safety of oral premedication in children with intracranial neurosurgical pathology before elective interventions in two modalities: a combination of midasolam 0.5 mg/kg body weight and ketamine 3 mg/kg (Group 1; n = 80) and midasolam 0.75 mg/kg only (Group 2; n = 20). The study was conducted in 100 children aged 2 to 10 years. Medical and sweet syrups were used to dilute the agents. In Groups 1 and 2, an effective anxiolytic effect was achieved in 92 and 80%, respectively. A sedative effect was also more pronounced in Group 1 children. The mean duration of effect achievement was 20 minutes. There were no serious complications when both regimens were used. Both oral premedication modes are considered to be effective and must be used for premedication in children with neurosurgical diseases.


Orthopaedic management of juvenile chronic arthritis (JCA).
Juvenile chronic arthritis is resulting in joint destruction and frequently also in growth disturbances. In less than 50% pain is the cause of functional disability. It is important for rheumasurgeons to differentiate between the three main types of JCA because of the different prognosis. It is also necessary to realize the involvement of all connective tissue and the multiple organ affections in this disease, especially the kidney-function and the hematopoietic system. The natural course of the disease is characterized by fluctuations between remissions and exacerbations, more irregularly than in the adult type of Rh.A. The good results of rheumasurgery are highly dependent on an interdisciplinary combined unit, preferably working "on the same floor" in daily cooperation. Special training in rheumasurgical operative technique is necessary. Prophylactic measures against joint contractures is of great importance and should be common knowledge of all members of the therapeutic team. During a period of 13 years 528 synovectomies were performed. In a controlled study of open knee-synovectomies, recurrence of the inflammation was rarely seen when a radical early synovectomy had been performed. The centralized unit with well experienced staff, who master the problems of anesthesia, medication, intraoperative blood-transfusion and physiotherapy pre- and post-operatively, may obtain good results in the management of this difficult disease.


Role of endotoxin in the response to experimentally induced bacteremia in chronically prepared rats.
Chronically prepared rats were injected intravenously with live Escherichia coli in doses from approximately 10(5) to approximately 10(9) colony-forming units (CFU). Significant dose-related increase in plasma adrenocorticotropin and corticosterone occurred after approximately 10(7) CFU. Fever occurred after approximately 10(7) CFU but not after approximately 10(9) CFU. These responses changed significantly but were not blocked completely when > 94% of the viable E. coli was removed from the inoculates. The remaining endotoxin activity in the inoculates resembled lipopolysaccharide (LPS) extracted from the same strain of E. coli on electrophoretic gels. Plasma endotoxin increased for > or = 240 min to 5.1 +/- 0.9 endotoxin units (EU)/ml after approximately 10(7) CFU and to 440 +/- 59 EU/ml after approximately 10(9) CFU. Endotoxin at approximately 10(9) CFU caused death within 24 h that was not predicted by the total activity of endotoxin that was injected. In contrast, extracted LPS with its strain and total activity matched to approximately 10(7) CFU mimicked the responses to this nonfatal dose. The total endotoxin activity of the injected bacteria appears to account for the effects of nonfatal doses of E. coli but not for the effects of fatal doses.


Role of TNF-alpha in high-dose antigen therapy in experimental autoimmune neuritis: inhibition of TNF-alpha by neutralizing antibodies reduces T-cell apoptosis and prevents liver necrosis.
TNF-alpha has been implicated as a potentially detrimental cytokine in autoimmune disorders of the nervous system and has been reported to be elevated in antigen-specific therapy of experimental autoimmune neuritis (EAN) in vivo. To investigate the role of TNF-alpha in EAN in rats that had been subjected to antigen-specific therapy with human P2 protein, animals were cotreated with an anti-TNF-alpha neutralizing antibody and the effects of the antibody on disease determined. Using this strategy in adoptive transfer (AT-) EAN, antigen-induced T-cell apoptosis in inflamed sciatic nerve and in liver was reduced to levels observed in control animals indicating that TNF-alpha mediates antigen-induced apoptosis of inflammatory T-cells. Focal liver necrosis, which had been observed in earlier studies after antigen therapy in AT-EAN, was prevented by passive immunization with neutralizing anti-TNF-alpha antibody. Unexpectedly, neutralization of TNF-alpha only partly abolished the protective effect of antigen therapy on the overall disease course. This may indicate that inhibition of TNF-alpha exerts beneficial effects other than through T-cell apoptosis, or that some of the benefit of antigen therapy is mediated by other pathways. These results indicate that secretion of TNF-alpha during antigen therapy has the dual potential to mediate beneficial apoptosis of inflammatory T-cells in the inflammatory lesion and to induce liver damage as a severe side effect.


Hypokalemic respiratory muscle paralysis following Strongyloides stercoralis hyperinfection: a case report.
Strongyloides stercoralis hyperinfection in a malnourished 6-year-old boy was characterized by severe diarrhea, dehydration and marked hypokalemia, followed by acute respiratory failure due to respiratory muscle paralysis, and cardiac arrest. He was resuscitated and maintained with positive pressure ventilation and intravenous infusion of potassium and gluco-saline solution. These measures produced recovery from the respiratory muscle paralysis. Stool examination revealed eggs and numerous larvae of S. stercoralis, and thiabendazole was initiated. He continued to have severe diarrhea and again developed marked hypokalemia with respiratory muscle paralysis, abdominal distention, and cardiac arrhythmias with cardiac arrest episodes. In spite of fluid and electrolyte replacement the patient died.


Lack of nosocomial spread of Varicella in a pediatric hospital with negative pressure ventilated patient rooms.
All patients at the new British Columbia's Children's Hospital with chickenpox or Herpes zoster are nursed with appropriate precautions in single-bed rooms provided with negative pressure ventilation. Over a period of 1 year, no nosocomial infections were detected on follow-up of 110 susceptible patients who had been on wards at the same time as six cases of chickenpox and one immunocompromised patient with cutaneous dissemination of Varicella zoster. In a preceding study at the previous hospital, with similar staff, control measures, and patient population, in an isolation facility without negative pressure ventilation, nosocomial infections occurred in seven out of 41 susceptible patients who were on the same ward as two patients with chickenpox. These findings suggest that a simple negative pressure ventilation system without air locks is a useful adjunct in the control of cutaneous Varicella infections.


Early and mid-term outcomes of surgery of the ascending aorta/arch: is there a relationship with caseload?
The relationship between caseload and early outcome remains a subject for debate in cardiac surgery. Surgery of the thoracic aorta is an area of specialist expertise within the adult cardiac surgical field. There is, however, a conflict between the concentration of expertise and the provision of effective emergency cover. This study evaluates the early and mid-term outcomes of patients undergoing surgery of the ascending aorta/aortic arch in a single institution and compares the results of a single higher volume surgeon with lower volume operators. From March 1992 till August 2003, 296 procedures were carried out on 291 patients (aged 17-80, median 62) who underwent operations for replacement of the ascending aorta/aortic arch. One hundred and thirty procedures were carried out by the higher volume surgeon and 160 by one of the six lower volume surgeons (range 10-57). Emergency operation was performed in 138 (47%) patients. One or more other associated cardiac procedures were carried out in 65 patients (22%). The overall early mortality was 37 (12.5%). After adjustment for baseline differences, era and surgical risk/complexity, the risk of in-hospital death was lower in the higher volume group, but not significantly so. For survival to 3 years the overall risk of death was significantly lower for patients in the higher volume group (hazard ratio 0.72; 95% CI 0.54-0.95) Apart from post-operative renal failure no other significant differences between the two groups were observed. Elective surgery of the ascending aorta/arch was associated with low mortality. Outcomes after emergency surgery conformed to contemporary expectations. Only limited differences were identified both with respect to the case profile and early clinical outcomes. Better outcomes in the mid-term in the higher volume group persisted despite adjustment for differences in caseload and are worthy of further study. We believe that these data support our hypothesis that dissemination of appropriate techniques among a group of surgeons represents the most practical method of service provision.