A phase 2 study of inotuzumab ozogamicin in patients with indolent B-cell non-Hodgkin lymphoma refractory to rituximab alone, rituximab and chemotherapy, or radioimmunotherapy.
This phase 2 study evaluated the efficacy and safety of inotuzumab ozogamicin (InO) in patients with indolent B-cell non-Hodgkin lymphoma (NHL) refractory to rituximab alone, rituximab plus chemotherapy or anti-CD20 radioimmunotherapy. Patients received InO 1·8 mg/m(2) intravenously on a 28-d cycle for a planned 4-8 cycles. The initial InO dose and schedule could be adjusted for tolerability and patients were allowed to receive 2 additional cycles (up to 8 total) after achieving a complete response (CR). The primary endpoint was overall response. Eighty-one patients were enrolled, among whom 48 (59%) received ≥3 InO cycles and 13 (16%) completed the treatment phase. The overall response rate was 67% (CR, 31%). Median (95% confidence interval) progression-free survival was 12·7 (8·9-26·9) months; median overall survival was not reached. Haematological adverse events (AEs) were common, particularly thrombocytopenia (74%) and neutropenia (56%). These were also the most common AEs leading to treatment discontinuation (37% and 11%, respectively); 58% of patients reported AEs leading to treatment discontinuation. InO demonstrated robust activity in these heavily pretreated patients, although treatment duration was limited by haematological toxicities. Additional studies may determine dosing regimens that allow for reduced toxicity.


Evaluation of zeolite-sand mixtures as reactive materials protecting groundwater at waste disposal sites.
To recognize properties of a mixture of Vistula sand (medium sand acc. to USCS) with Slovak zeolite as reactive materials suitable for permeable reactive barriers proposed for protection of groundwater environment in vicinity of old landfills comprehensive laboratory investigations were performed. The present study investigates the removal of contaminants specific for landfill leachates onto zeolite-sand mixtures containing 20%, 50% and 80% of zeolite (ZS20, ZS50 and ZS80). Taking into account the results of batch tests it was concluded that the Langmuir isotherm best fitted the data. It was observed that the presence of ammonium, calcium and magnesium decreases the removal efficiency of copper by 32%. Column tests of contaminant migration through the attenuation zone of the reactive materials were interpreted using the software package CXTFIT, which solves a one-dimensional advection-dispersion equation. Column test results also indicate the strong influence of the presence of interfering substances on copper immobilisation; dynamic sorption capacities decrees twofold. Throughout the landfill leachate flow through ZS80 sample, a constant reduction of NH4+ (at 100%), K+ (at 93%) and Fe(total) (at an average of 86%) were observed. There was no reduction in chemical oxygen demand and biochemical oxygen demand.


Anti-tumor effects and potential therapeutic response biomarkers in α-emitting meta-211At-astato-benzylguanidine therapy for malignant pheochromocytoma explored by RNA-sequencing.
Targeted α-particle therapy is a promising option for patients with malignant pheochromocytoma. Recent observations regarding meta-211At-astato-benzylguanidine (211At-MABG) in a pheochromocytoma mouse model showed a strong anti-tumor effect, though the molecular mechanism remains elusive. Here, we present the first comprehensive RNA-sequencing (RNA-seq) data for pheochromocytoma cells based on in vitro211At-MABG administration experiments. Key genes and pathways in the tumor α-particle radiation response are also examined to obtain potential response biomarkers. Methods: We evaluated genome-wide transcriptional alterations in the rat pheochromocytoma cell line PC12 at 3, 6, and 12 h after 211At-MABG treatment; a control experiment using 60Co γ-ray irradiation was carried out to highlight 211At-MABG-specific gene expression. For comparisons, 10% and 80% iso-survival doses (0.8 and 0.1 kBq/mL for 211At-MABG and 10 and 1 Gy for 60Co γ-rays) were used. Results: Enrichment analysis of differentially expressed genes (DEGs) and analysis of the gene expression profiles of cell cycle checkpoints revealed similar modes of cell death via the p53-p21 signaling pathway after 211At-MABG treatment and γ-ray irradiation. The top list of ranked DEGs demonstrated the expression of key genes on the decrease in the survival following 211At-MABG exposure, and four potential genes (Mien1, Otub1, Vdac1 and Vegfa genes) of 211At-MABG therapy. Western blot analysis indicated increased expression of TSPO in 211At-MABG-treated cells, suggesting its potential as a PET imaging probe. Conclusion: Comprehensive RNA-seq revealed contrasting cellular responses to γ-ray and α-particle therapy, leading to the identification of four potential candidate genes that may serve as molecular imaging and 211At-MABG therapy targets.


Free radicals in ischemic and reperfusion myocardial injury: is this the time for clinical trials?
An increasing number of studies in cell cultures, isolated perfused hearts, and intact animal preparations purport to show the significance of free radical production in the pathophysiology of myocardial injury and necrosis of ischemic and reoxygenated or reperfused tissue. Additionally an impressive array of therapeutic approaches has been developed to interfere with the generation of free radicals and to thereby salvage jeopardized myocardium. Based on these data clinical trials are now being conducted or planned. However, a critical examination of the experimental data raises questions that undermine total acceptance of the conclusions of the investigators and the enthusiastic extrapolations to the clinical arena. Issues such as endotoxin contamination of infused free radical scavenger enzymes, incorrect dosing, possibly improper selection of control conditions, species specificity, inadequate consideration of the dependence of infarct size on coronary collateral blood flow, and the possibly transient nature of any benefits of anti-free radical interventions cast doubt on some of the accumulated data and their general applicability to humans. These issues must be satisfactorily addressed before this experimental approach can be fully embraced by the clinical cardiologist. Additionally, uniformity in the experimental model and conditions might eliminate some of the confusion and make it easier to compare the results of different studies. Free radical mechanisms may be important in the heart, but more data are needed to rigorously document an unequivocal effect of therapeutic interventions in experimental models before clinical trials are appropriate.


Connective tissue growth factor is expressed in malignant cells of Hodgkin lymphoma but not in other mature B-cell lymphomas.
Connective tissue growth factor (CTGF) has a major role in development of fibrosis and in the wound-healing process. Microarray analysis of 44 classical Hodgkin lymphoma (cHL) samples showed higher CTGF messenger RNA expression in the nodular sclerosis (NS) than in the mixed cellularity (MC) subtype. When analyzed by immunohistochemical analysis, Hodgkin-Reed-Sternberg (H-RS) cells and macrophages in 23 cHLs and "popcorn" cells in 2 nodular lymphocyte predominant Hodgkin lymphomas showed expression of CTGF protein correlating with the extent of fibrosis. In NS, CTGF was also expressed in fibroblasts and occasional lymphocytes. Malignant cells in 32 samples of various non-Hodgkin lymphomas were negative for CTGF. A staining pattern of stromal cells similar to that of NS cHL was seen in anaplastic large cell lymphoma. Macrophages stained positively in Burkitt lymphomas and in some mantle cell lymphomas. The high occurrence of fibrosis in cHL may be related to CTGF expression by malignant H-RS cells.


Neuroanatomical Basis of Individuality in Muscle Tuning Function: Neural Correlates of Muscle Tuning.
In a conventional view of motor control, the human brain might employ an optimization principle that leads a stereotypical motor behavior which we observe as an averaged behavioral data over subjects. In this scenario, the inter-individual motor variability is considered as an observation noise. Here, we challenged this view. We considered a motor control task where the human participants manipulated arm force by coordinating shoulder and elbow torques and investigated the muscle-tuning function that represents how the brain distributed the ideal joint torques to multiple muscles. In the experimental data, we observed large inter-individual variability in the profile of a muscle-tuning function. This contradicts with a well-established optimization theory that is based on minimization of muscle energy consumption and minimization of motor variability. We then hypothesized the inter-subject differences in the structure of the motor cortical areas might be the source of the across-subjects variability of the motor behavior. This was supported by a voxel-based morphometry analysis of magnetic resonance imaging; The inter-individual variability of the muscle tuning profile was correlated with that of the gray matter volume in the premotor cortex which is ipsilateral to the used arm (i.e., right hemisphere for the right arm). This study suggests that motor individuality may originate from inter-individual variation in the cortical structure.


Shoulder appearances at MR imaging in long-term dialysis recipients.
To examine shoulder appearances at magnetic resonance (MR) imaging in long-term dialysis recipients. Twenty-two chronic dialysis recipients underwent 1.0-T MR imaging with a combination of T1-, T2-, and T2*-weighted sequences. Rotator cuff tendon thickening was graded as present or absent by a musculoskeletal radiologist, who also measured the supraspinatus and subscapularis tendon thicknesses with electronic calipers. The long-axis dimension and location of focal osseous lesions, in addition to their T1, T2, and T2* signal intensities, were noted. Supraspinatus (n = 9) and subscapularis (n = 10) tendon thickening was frequently observed. Six (27%) of the 22 patients had combined thickening of the supraspinatus and subscapularis tendons without substantial involvement of the infraspinatus or teres minor tendons. These patients had undergone dialysis longer (median, 19.2 years; range, 16.3-22.8 years) than had the other patients (median, 11.7 years; range, 5.8-19.3 years; P: =.004). The 29 intraosseous lesions had high, intermediate, and low T2 signal intensity in six (21%), nine (31%), and 14 (48%) instances, respectively. Supraspinatus and/or subscapularis tendon thickening is common in chronic dialysis recipients. Bone lesions in such patients are of variable T2 signal intensity and usually subchondral or adjacent to the greater tuberosity.


Climate change adaptation and the role of fuel subsidies: An empirical bio-economic modeling study for an artisanal open-access fishery.
Climate change can severely impact artisanal fisheries and affect the role they play in food security. We study climate change effects on the triple bottom line of ecological productivity, fishers' incomes, and fish consumption for an artisanal open-access fishery. We develop and apply an empirical, stochastic bio-economic model for the Senegalese artisanal purse seine fishery on small pelagic fish and compare the simulated fishery's development using four climate projections and two policy scenarios. We find that economic processes of adaptation may amplify the effects of climate variations. The regions' catch potential increases with climate change, induced by stock distribution changes. However, this outcome escalates over-fishing, whose effects outpace the incipiently favorable climate change effects under three of the four climate projections. Without policy action, the fishery is estimated to collapse in 2030-2035 on average over 1000 runs. We propose an easily implementable and overall welfare-increasing intervention: reduction of fuel subsidies. If fuel subsidies were abolished, ecological sustainability as well as the fishery's welfare contribution would increase regardless of the climate projection.


Frequent inactivation of PTEN by promoter hypermethylation in microsatellite instability-high sporadic colorectal cancers.
Loss of PTEN tumor suppressor function is observed in tumors of breast, prostate, thyroid, and endometrial origin. Allelic losses in the proximity of the PTEN locus (10q23) also occur in sporadic colorectal cancers (CRCs), but biallelic inactivation of this site has not been frequently demonstrated. We hypothesized that alternative mechanisms of PTEN allelic inactivation, such as promoter hypermethylation, might be operative in CRC and that PTEN inactivation may be related to recognized forms of genomic instability. We characterized a cohort of 273 sporadic CRCs by determining their microsatellite instability (MSI) status. Of these, 146 cancers were examined for PTEN promoter methylation by methylation-specific PCR. Mutations at the poly(A)6 repeat sequences in PTEN exons 7 and 8 and deletions at the 10q23 locus were also identified using microsatellite analysis. The presence of PTEN protein was determined by immunostaining, and the results were correlated with the promoter methylation status. We observed that PTEN promoter hypermethylation was a frequent occurrence in MSI-high (MSI-H) tumors (19.1% of MSI-H versus 2.2% of MSI-low/microsatellite stable tumors; P = 0.002). A PTEN mutation or a deletion event was present in 60% of the tumors with promoter region hypermethylation. Hypermethylation of the PTEN promoter correlated significantly with either decreased or complete loss of PTEN protein expression (P = 0.004). This is the first demonstration of PTEN inactivation as a result of promoter hypermethylation in MSI-H sporadic CRCs. These data suggest that this silencing mechanism plays a major role in PTEN inactivation and, in colon cancer, may be more important than either allelic losses or inactivating mutations. The significant correlation of PTEN hypermethylation with MSI-H tumors further suggests that PTEN is an additional important "target" of methylation along with the hMLH1 gene in the evolution of MSI-H CRCs and also confers the "second hit" in the biallelic inactivation mechanism for some proportion of tumors.


The specificity of the search template.
When searching for a target object, observers use an internal representation of the target's appearance as a search template. This study used naturalistic stimuli to examine the specificity of this template. Observers first learned several name-image pairs; they then participated in a search experiment in which the names served as cues and the images served as targets. To test whether the observers searched for the targets using an exact image template, we included targets that were transformations of the studied image and targets that belonged to the same subordinate-level category as the studied image. The same stimuli were also used in a search experiment involving image cues. The name cue and image cue experiments produced different patterns of results. Unlike image cues, name cues produced similar benefits for transformations of the studied images as for the studied images themselves. Also unlike image cues, names cues produced no benefit for members of the same subordinate-level category as the studied image. These results suggest that when observers are trained on an image, they develop a search template that is relatively specific for the image but still tolerant to changes in scale and orientation.


Are posture data from simulated tasks representative of field conditions? Case study for overhead electric utility workers.
Many ergonomics studies are conducted in laboratory-simulated work environments to assess risks for the development of musculoskeletal disorders under more controlled conditions. However, the simulated conditions could be of questionable validity with respect to reproduction of field conditions involving risk factors. The objective of this study was to verify whether the postures recorded for neck extension/flexion and upper arm elevation of overhead electric utility workers in a simulated environment were similar to those recorded in a field environment. Of the three frequently performed tasks analysed, two presented similar exposure in both conditions. However, differences were identified for a more complex task (relay replacement). These results suggest that simulated tasks may be more representative for more standardised tasks. This may indicate that researchers investigating risks should avoid simplifying complex tasks when reproducing field posture exposure in laboratories, since omitting extra subtasks may lead to an inaccurate reproduction of field exposure. Practitioner Summary: Studies comparing results from field and simulated environments are necessary to evaluate to what degree postural exposure reproduced in laboratory is representative of the exposure occurring in real work situations. This is particularly relevant for tasks involving training in simulated environment due to safety constraints.


Sequencing batch reactor process for the removal of nitrogen from anaerobically treated domestic wastewater.
This work presents the performance of a sequencing batch reactor (SBR) system used as a means of removing nitrogen from domestic wastewater containing a low chemical oxygen demand (COD) to nitrogen ratio due to pre-treatment with an anaerobic reactor. The aim of the work was to determine the feasibility of this system for the removal of nitrogen from the domestic wastewater. An SBR with a working volume of 5 L was investigated at different cycle times of 12, 8 and 6 h, at 18 °C. The efficiency of the SBR varied together with the duration of the cycle, where the optimum performance was seen in the 6 h cycle with the anoxic-aerobic-anoxic sequence. Due to the low quantity of organic matter present in the domestic wastewater after the anaerobic treatment, an additional supply of external carbon was necessary before the second anoxic stage. The removal efficiencies obtained were: 98% for total Kjeldahl nitrogen, 84% for total nitrogen and 77% for soluble COD. The reactor was thus shown to be viable, and it was concluded that this process may be successfully applied as a post-treatment for the removal of nitrogen from anaerobically treated domestic wastewater.


The overarm-throwing pattern among U-14 ASA female softball players: a comparative study of gender, culture, and experience.
A developmental description of overarm-throwing characteristics of U-14female ASA softballplayers is presented here. Comparisons were made between these athletes and teens of similar age in the United States (Runion, Roberton, & Langendorfe 2003) and in Germany (Ehl, Roberton, & Langendorfer, 2005). A majority of the softball players demonstrated the most advanced developmental levels for the backswing foot, humerus, and forearm components. Seventy percent (n = 26) demonstrated a Level 2 trunk, while 30% (n = 11) demonstrated a differentiated trunk, Level 3. The average throwing velocity was 62.58 ft/s (19.07 m/s). Comparisons were made to girls with less throwing experience in the United States (Runion et al.) and Germany (Ehl et al.); the softball players threw with greater velocity (9.12ft/s [2.77 m/s]and 15.89 ft/s [4.84 m/s], respectively) and demonstrated superior trunk, humerus, and forearm actions. The boys from both countries threw the ball faster than the softball players. Significant chi-square analyses found the softball players had superior humeral actions (Level 3) compared to the German boys and superior forearm actions (Level 3) compared to both groups of bays. Experience seems to have an impact when comparisons among girls are made; yet, explanations for differences seen between boys and girls still remain unclear and warrant further research.


Angular dependence of the interaction energy between the N lone pair of amines and a proton: relevance to drug-receptor systems.
Ab initio molecular orbital calculations with a 4-31G basis set have been performed to study the angular dependence of the interaction energy between a lone electron pair of nitrogen and a proton. In this study ammonia and trimethylamine were used as models of biologically active amines. A proton was used as a model of an electrophilic site at the receptor. Results obtained confirm previous indications that the energy required to bend the proton from the lone pair direction decreases markedly as the two species are further separated from one another. Implications regarding the interactions of drugs and hormones at specific receptors are discussed.


Nutritional status of under-five children living in an informal urban settlement in Nairobi, Kenya.
Malnutrition in sub-Saharan Africa contributes to high rates of childhood morbidity and mortality. However, little information on the nutritional status of children is available from informal settlements. During the period of post-election violence in Kenya during December 2007-March 2008, food shortages were widespread within informal settlements in Nairobi. To investigate whether food insecurity due to post-election violence resulted in high prevalence of acute and chronic malnutrition in children, a nutritional survey was undertaken among children aged 6-59 months within two villages in Kibera, where the Kenya Medical Research Institute/Centers for Disease Control and Prevention conducts population-based surveillance for infectious disease syndromes. During 25 March-4 April 2008, a structured questionnaire was administered to caregivers of 1,310 children identified through surveillance system databases to obtain information on household demographics, food availability, and child-feeding practices. Anthropometric measurements were recorded on all participating children. Indices were reported in z-scores and compared with the World Health Organization (WHO) 2005 reference population to determine the nutritional status of children. Data were analyzed using the Anthro software of WHO and the SAS. Stunting was found in 47.0% of the children; 11.8% were underweight, and 2.6% were wasted. Severe stunting was found in 23.4% of the children; severe underweight in 3.1%, and severe wasting in 0.6%. Children aged 36-47 months had the highest prevalence (58.0%) of stunting while the highest prevalence (4.1%) of wasting was in children aged 6-11 months. Boys were more stunted than girls (p < 0.01), and older children were significantly (p < 0.0001) stunted compared to younger children. In the third year of life, girls were more likely than boys to be wasted (p < 0.01). The high prevalence of chronic malnutrition suggests that stunting is a sustained problem within this urban informal settlement, not specifically resulting from the relatively brief political crisis. The predominance of stunting in older children indicates failure in growth and development during the first two years of life. Food programmes in Kenya have traditionally focused on rural areas and refugee camps. The findings of the study suggest that tackling childhood stunting is a high priority, and there should be fostered efforts to ensure that malnutrition-prevention strategies include the urban poor.


The effect of proximity to hurricanes Katrina and Rita on subsequent hurricane outlook and optimistic bias.
This study evaluated how individuals living on the Gulf Coast perceived hurricane risk after Hurricanes Katrina and Rita. It was hypothesized that hurricane outlook and optimistic bias for hurricane risk would be associated positively with distance from the Katrina-Rita landfall (more optimism at greater distance), controlling for historically based hurricane risk and county population density, demographics, individual hurricane experience, and dispositional optimism. Data were collected in January 2006 through a mail survey sent to 1,375 households in 41 counties on the coast (n = 824, 60% response). The analysis used hierarchal regression to test hypotheses. Hurricane history and population density had no effect on outlook; individuals who were male, older, and with higher household incomes were associated with lower risk perception; individual hurricane experience and personal impacts from Katrina and Rita predicted greater risk perception; greater dispositional optimism predicted more optimistic outlook; distance had a small effect but predicted less optimistic outlook at greater distance (model R(2) = 0.21). The model for optimistic bias had fewer effects: age and community tenure were significant; dispositional optimism had a positive effect on optimistic bias; distance variables were not significant (model R(2) = 0.05). The study shows that an existing measure of hurricane outlook has utility, hurricane outlook appears to be a unique concept from hurricane optimistic bias, and proximity has at most small effects. Future extension of this research will include improved conceptualization and measurement of hurricane risk perception and will bring to focus several concepts involving risk communication.


Heparan sulfate chains of perlecan are indispensable in the lens capsule but not in the kidney.
Mice lacking exon 3 of perlecan (Hspg2) gene were generated by gene targeting. Exon deletion does not alter the expression or the reading frame but causes loss of attachment sites for three heparan sulfate (HS) side chains. Hspg2(Delta 3 / Delta 3) mice are viable and fertile but have small eyes. Apoptosis and leakage of cellular material through the lens capsule are observed in neonatal lenses, and lenses degenerate within 3 weeks of birth. Electron microscopy revealed altered structure of the lens capsule through which cells had formed extensions. No kidney malfunction, such as protein uria, was detected in Hspg2(Delta 3 / Delta 3) mutant mice, nor were ultrastructural changes observed in the glomerular basement membranes (BMs). To achieve further depletion in the HS content of the BMs, Hspg2(Delta 3 / Delta 3) mice were bred with collagen XVIII null mice. Lens defects were more severe in the newborn Col18a1(-/-) x Hspg2(Delta 3 / Delta 3) mice and degeneration proceeded faster than in Hspg2(Delta 3 / Delta 3) mice. The results suggest that in the lens capsule, HS chains have a structural function and are essential in the insulation of the lens from its environment and in regulation of incoming signals.


Using High-Dimensional Approaches to Probe Monocytes and Macrophages in Cardiovascular Disease.
High dimensional approaches that characterize single cells at unprecedented depth have helped uncover unappreciated heterogeneity, a better understanding of myeloid cell origins, developmental relationships and functions. These advancements are particularly important in cardiovascular disease, which remains the leading cause of death worldwide. Gradual, monocyte-dependent inflammatory processes, such as the development of atherosclerotic plaque within arterial vessels, contrasts with the robust acute response within the myocardium that occurs when a vessel is occluded. Monocytes and macrophages differentially contribute to tissue injury, repair and regeneration in these contexts, yet many questions remain about which myeloid cell types are involved in a coordinated, organ-level sterile inflammatory response. Single cell RNA sequencing, combined with functional analyses have demonstrated that at least three populations of resident cardiac macrophages exist, and after tissue injury, there is significant diversification of the tissue macrophage pool driven by recruited monocytes. While these studies have provided important insights, they raise many new questions and avenues for future exploration. For example, how do transcriptionally defined sub-populations of cardiac macrophages relate to each other? Are they different activation states along a pre-defined trajectory of macrophage differentiation or do local microenvironments drive newly recruited monocytes into distinct functions? The answers to these questions will require integration of high-dimensional approaches into biologically relevant in vivo experimental systems to ensure the predicted heterogeneity possess a functional outcome.


Rationale and design of the LosmApimod To Inhibit p38 MAP kinase as a TherapeUtic target and moDify outcomes after an acute coronary syndromE trial.
p38 mitogen-activated protein kinase (MAPK) mediates cytokine production and amplification of the inflammatory cascade. Through inhibition of p38 MAPK, losmapimod appears to attenuate the inflammatory response in the vascular wall and thus may help stabilize plaques. The LATITUDE-TIMI 60 trial is a randomized, double-blind, placebo-controlled, parallel-group, multicenter study planned to be conducted in a 3-stage design. Overall, the trial is designed to include 25,500 patients hospitalized with non-ST-elevation or ST-elevation myocardial infarction (MI) randomized to oral losmapimod (7.5 mg twice daily) versus matching placebo. Part A consists of a leading cohort (n = 3,500) that will provide an initial assessment of safety and exploratory efficacy before progressing to part B. Part B (n = ~22,000) of the study is event driven and will provide the primary assessment of efficacy. An independent safety review will be conducted after 3,500 patients in part B1 to determine whether a more focused schedule of clinic visits and laboratory assessments can be implemented (part B2). All patients are to be treated with study drug until week 12 and followed up until week 24. The primary end point is the composite of cardiovascular death, MI, or severe recurrent ischemia requiring urgent coronary revascularization. The key secondary end point is the composite of cardiovascular death or MI. The trial is designed to provide ≥90% power for the primary end point. The LATITUDE-TIMI 60 trial will determine the efficacy and safety of short-term p38 MAPK inhibition with losmapimod in acute MI. The trial design adopts a stepwise approach to decision making and collection of data.


Glucosamine/L-lactide copolymers as potential carriers for the development of a sustained rifampicin release system using Mycobacterium smegmatis as a tuberculosis model.
The present study aims at developing a new, ultrafine particle-based efficient antibiotic delivery system for the treatment of tuberculosis. The carrier material to make the rifampicin (RIF)-loaded particles is a low molecular weight star-shaped polymer produced from glucosamine (core building unit) and L-lactide (GluN-LLA). Particles were made via electrohydrodynamic atomization. Prolonged release (for up to 14 days) of RIF from these particles is reported. Drug release data fits the Korsmeyer-Peppas equation, which suggests the occurrence of a modified diffusion-controlled RIF release mechanism in vitro and is also supported by differential scanning calorimetry and drug leaching tests. Cytotoxicity tests on Mycobacterium smegmatis showed that antibiotic-free GluN-LLA and polylactides (PLA) particles (reference materials) did not show any significant anti-bacterial activity. The minimum inhibitory concentration and minimum bactericidal concentration values obtained for RIF-loaded particles showed 2- to 4-fold improvements in the anti-bacterial activity relative to the free drug. Cytotoxicity tests on macrophages indicated that cell death correlates with an increase of particle concentration but is not significantly affected by material type or particle size. Confocal microscopy was used to track internalization and localization of particles in the macrophages. The uptake of GluN-LLA particles is higher than those of their PLA counterparts. In addition, after phagocytosis, the GluN-LLA particles stayed in the cytoplasm and showed favorable long-term drug release behavior, which facilitated the killing of intracellular bacteria when compared to free RIF. The present studies suggest that these drug carrier materials are potentially very attractive candidates for the development of high-payload, sustained-release antibiotic/resorbable polymer particle systems for treating bacterial lung infections.


Rheb, an activator of target of rapamycin, in the blackback land crab, Gecarcinus lateralis: cloning and effects of molting and unweighting on expression in skeletal muscle.
Molt-induced claw muscle atrophy in decapod crustaceans facilitates exuviation and is coordinated by ecdysteroid hormones. There is a 4-fold reduction in mass accompanied by remodeling of the contractile apparatus, which is associated with an 11-fold increase in myofibrillar protein synthesis by the end of the premolt period. Loss of a walking limb or claw causes a loss of mass in the associated thoracic musculature; this unweighting atrophy occurs in intermolt and is ecdysteroid independent. Myostatin (Mstn) is a negative regulator of muscle growth in mammals; it suppresses protein synthesis, in part, by inhibiting the insulin/metazoan target of rapamycin (mTOR) signaling pathway. Signaling via mTOR activates translation by phosphorylating ribosomal S6 kinase (s6k) and 4E-binding protein 1. Rheb (Ras homolog enriched in brain), a GTP-binding protein, is a key activator of mTOR and is inhibited by Rheb-GTPase-activating protein (GAP). Akt protein kinase inactivates Rheb-GAP, thus slowing Rheb-GTPase activity and maintaining mTOR in the active state. We hypothesized that the large increase in global protein synthesis in claw muscle was due to regulation of mTOR activity by ecdysteroids, caused either directly or indirectly via Mstn. In the blackback land crab, Gecarcinus lateralis, a Mstn-like gene (Gl-Mstn) is downregulated as much as 17-fold in claw muscle during premolt and upregulated 3-fold in unweighted thoracic muscle during intermolt. Gl-Mstn expression in claw muscle is negatively correlated with hemolymph ecdysteroid level. Full-length cDNAs encoding Rheb orthologs from three crustacean species (G. lateralis, Carcinus maenas and Homarus americanus), as well as partial cDNAs encoding Akt (Gl-Akt), mTOR (Gl-mTOR) and s6k (Gl-s6k) from G. lateralis, were cloned. The effects of molting on insulin/mTOR signaling components were quantified in claw closer, weighted thoracic and unweighted thoracic muscles using quantitative polymerase chain reaction. Gl-Rheb mRNA levels increased 3.4-fold and 3.9-fold during premolt in claw muscles from animals induced to molt by eyestalk ablation (ESA) and multiple leg autotomy (MLA), respectively, and mRNA levels were positively correlated with hemolymph ecdysteroids. There was little or no effect of molting on Gl-Rheb expression in weighted thoracic muscle and no correlation of Gl-Rheb mRNA with ecdysteroid titer. There were significant changes in Gl-Akt, Gl-mTOR and Gl-s6k expression with molt stage. These changes were transient and were not correlated with hemolymph ecdysteroids. The two muscles differed in terms of the relationship between Gl-Rheb and Gl-Mstn expression. In thoracic muscle, Gl-Rheb mRNA was positively correlated with Gl-Mstn mRNA in both ESA and MLA animals. By contrast, Gl-Rheb mRNA in claw muscle was negatively correlated with Gl-Mstn mRNA in ESA animals, and no correlation was observed in MLA animals. Unweighting increased Gl-Rheb expression in thoracic muscle at all molt stages; the greatest difference (2.2-fold) was observed in intermolt animals. There was also a 1.3-fold increase in Gl-s6k mRNA level in unweighted thoracic muscle. These data indicate that the mTOR pathway is upregulated in atrophic muscles. Gl-Rheb, in particular, appears to play a role in the molt-induced increase in protein synthesis in the claw muscle.


Division of Labor among Primitively Social Bees.
Australian bees of the genera Allodapula and Exoneura which commonly live in colonies of two to several individuals exhibit a division of labor among the adult females. Most foraging individuals are unfertilized and have slender ovaries, yet gather pollen and carry it to the nest; in contrast, fertilized egg-laying bees are not commonly foragers. Such castes are functionally similar to workers and queens found in some members of two other families of bees and represent a noteworthy example of physiological and behavioral parallelism in which the activities of different individuals are coordinated to form a functional unit.


Evaluation of the Safety of Everolimus-Eluting Bioresorbable Vascular Scaffold (BVS) Implantation in Patients With Chronic Total Coronary Occlusions: Acute Procedural and Short-Term Clinical Results.
The bioresorbable vascular scaffold (BVS) (Abbott Vascular) provides temporary scaffolding while eluting everolimus. There are limited data on its use in daily practice, especially in patients with stable angina pectoris referred for elective percutaneous coronary intervention (PCI) of chronic total occlusions (CTOs). The current study aims to investigate the safety and efficacy of BVS implantation in a selected patient cohort with CTO. A total of 70 consecutive patients, who underwent successful recanalization of CTO with BVS between September 13, 2012 and September 20, 2014 in three cardiac centers (Department of Cardiology, Bezmialem Vakif University, Istanbul, Turkey; Department of Interventional Cardiology, San Raffaele Hospital, Milan, Italy; and EMO-GVM Centro Cuore Columbus, Milan, Italy) were included in this CTO registry. Endpoints analyzed included: (1) the composite of all-cause death and non-fatal myocardial infarction (MI); and (2) the composite safety endpoint of major adverse cardiovascular events (MACEs), including death, MI and symptom-driven target lesion revascularization (TLR). Clinical data were obtained for 70 patients (mean age, 56.9 ± 9.4 years; 90.0% male) with a total number of 76 CTOs. At a median follow-up of 11.0 months (interquartile range, 7-18 months), both MACE and TLR rates were 4.3%. Two patients suffered from ischemia-driven TLR (1 patient at 6 months and 1 patient at 9 months after implantation). No death, MI, or stent thrombosis was observed during the follow-up period. Treatment of CTOs with BVS seems to be safe and effective, with a high technical success rate and acceptable MACE at short-term follow-up.


Challenges associated with the implementation of the nursing process: A systematic review.
Nursing process is a scientific approach in the provision of qualified nursing cares. However, in practice, the implementation of this process is faced with numerous challenges. With the knowledge of the challenges associated with the implementation of the nursing process, the nursing processes can be developed appropriately. Due to the lack of comprehensive information on this subject, the current study was carried out to assess the key challenges associated with the implementation of the nursing process. To achieve and review related studies on this field, databases of Iran medix, SID, Magiran, PUBMED, Google scholar, and Proquest were assessed using the main keywords of nursing process and nursing process systematic review. The articles were retrieved in three steps including searching by keywords, review of the proceedings based on inclusion criteria, and final retrieval and assessment of available full texts. Systematic assessment of the articles showed different challenges in implementation of the nursing process. Intangible understanding of the concept of nursing process, different views of the process, lack of knowledge and awareness among nurses related to the execution of process, supports of managing systems, and problems related to recording the nursing process were the main challenges that were extracted from review of literature. On systematically reviewing the literature, intangible understanding of the concept of nursing process has been identified as the main challenge in nursing process. To achieve the best strategy to minimize the challenge, in addition to preparing facilitators for implementation of nursing process, intangible understanding of the concept of nursing process, different views of the process, and forming teams of experts in nursing education are recommended for internalizing the nursing process among nurses.


Prevalence and treatment of neurological and psychiatric disorders among tertiary hospitals in Pakistan; findings and implications.
Mental health and neurological disorders are prevalent in Pakistan. However, there are considerable concerns with their management due to issues of access, availability of trained personnel and stigma alongside paucity of such data. Consequently, there is a need to document current treatment approaches starting with tertiary hospitals in Pakistan where patients with more severe mental and neurological disorders are typically treated. Subsequently, use the findings to help direct future policies and initiatives. Multi-centered, cross-sectional, prospective study principally evaluating current medicine usage among patients attending tertiary hospitals in Pakistan with psychiatric and neurological disorders. In addition, possible factors contributing to the prevalence of these disorders in this population to help with future care. All 23 tertiary care hospitals in the ten major Districts in Pakistan were included, which cover 75% of the population. 57,664 patients were evaluated of which 35.3% were females. Both females and males had multiple brain disorders and multiple co-morbidities. Schizophrenia was the most prevalent disorder overall among both females (25.2%) and males (30.4%). A median of six medicines were prescribed per patient, with antipsychotics and antidepressants the most prescribed medicines. Clozapine was the most prescribed medicine in males (12.25%) and females (11.83%) including for psychiatric disorders, with sodium valproate the most prescribed medicine in epilepsy in males (42.44% of all anti-epileptic medicines) as well as females (46.38%). There was a greater prevalence of both disorders among the lower classes. A greater prevalence of schizophrenia was seen in patients abusing alcohol and smokers. The divorce rate was higher among the studied patients and the prevalence of depression was higher among the widowed population. There were concerns with the quality of prescribing including the extent of polypharmacy as well as possible overuse of clozapine especially in patients with epilepsy, both of which need addressing.


Were early pterosaurs inept terrestrial locomotors?
Pterodactyloid pterosaurs are widely interpreted as terrestrially competent, erect-limbed quadrupeds, but the terrestrial capabilities of non-pterodactyloids are largely thought to have been poor. This is commonly justified by the absence of a non-pterodactyloid footprint record, suggestions that the expansive uropatagia common to early pterosaurs would restrict hindlimb motion in walking or running, and the presence of sprawling forelimbs in some species. Here, these arguments are re-visited and mostly found problematic. Restriction of limb mobility is not a problem faced by extant animals with extensive fight membranes, including species which routinely utilise terrestrial locomotion. The absence of non-pterodactyloid footprints is not necessarily tied to functional or biomechanical constraints. As with other fully terrestrial clades with poor ichnological records, biases in behaviour, preservation, sampling and interpretation likely contribute to the deficit of early pterosaur ichnites. Suggestions that non-pterodactyloids have slender, mechanically weak limbs are demonstrably countered by the proportionally long and robust limbs of many Triassic and Jurassic species. Novel assessments of pterosaur forelimb anatomies conflict with notions that all non-pterodactyloids were obligated to sprawling forelimb postures. Sprawling forelimbs seem appropriate for species with ventrally-restricted glenoid articulations (seemingly occurring in rhamphorhynchines and campylognathoidids). However, some early pterosaurs, such as Dimorphodon macronyx and wukongopterids, have glenoid arthrologies which are not ventrally restricted, and their distal humeri resemble those of pterodactyloids. It seems fully erect forelimb stances were possible in these pterosaurs, and may be probable given proposed correlation between pterodactyloid-like distal humeral morphology and forces incurred through erect forelimb postures. Further indications of terrestrial habits include antungual sesamoids, which occur in the manus and pes anatomy of many early pterosaur species, and only occur elsewhere in terrestrial reptiles, possibly developing through frequent interactions of large claws with firm substrates. It is argued that characteristics possibly associated with terrestriality are deeply nested within Pterosauria and not restricted to Pterodactyloidea as previously thought, and that pterodactyloid-like levels of terrestrial competency may have been possible in at least some early pterosaurs.


Liposomal bupivacaine in forefoot surgery.
Liposomal bupivacaine is increasingly being utilized in attempts to provide prolonged local analgesia in the immediate postoperative period. The purpose of this study was to quantify the reduction in opioid consumption as well as postoperative pain scores in the postoperative period when liposomal bupivacaine is used at the conclusion of forefoot surgery. This was a prospective therapeutic cohort study with 20 patients receiving liposomal bupivacaine at the conclusion of their forefoot procedure in addition to our routine multimodal analgesic protocol and 20 patients as the control without the addition of liposomal bupivacaine. Pain scores, number of narcotic pills consumed on postoperative days 1 through 4, need for refill, time to first refill and wound complications were evaluated. Mean number of narcotic pills consumed on postoperative day 1 (1.4 vs 3.6, P = .002) and day 2 (1.8 vs 3.6, P = .021) was significantly lower for the liposomal bupivacaine group than the control group. Daily pain scores were lower for those patients who received liposomal bupivacaine on postoperative days 1 through 4, although this was not statistically significant. Fewer patients required medication refills in the liposomal bupivacaine group compared to the control group, however this was not statistically significant (3 vs 7, P = .273). There was no increase in wound complications in the liposomal bupivacaine group when compared to the control (1 vs 4). Liposomal bupivacaine appears to be a useful adjunct in the treatment of postoperative pain when used as a part of a multimodal analgesic regimen in patients undergoing forefoot surgery. Level II, prospective comparative study.


Nutritional metabolic bone disease in juvenile veiled chameleons (Chamaeleo calyptratus) and its prevention.
Nutritional metabolic bone disease (NMBD) is one of the most frequently observed pathological conditions in herpetoculture. To develop guidelines for NMBD prevention in growing veiled chameleons (Chamaeleo calyptratus), 56 hatchlings were divided into 6 groups [group UV, with UVB exposure; group No: no supplements; group CaAUV: with calcium (Ca), vitamin A, UVB; group CaA: with Ca, vitamin A; group CaADUV: with Ca, vitamin A, cholecalciferol, UVB; and group CaAD, with Ca, vitamin A, cholecalciferol] and reared for 6 mo on locust-based diets. The nutrient composition of the locusts' diet and the locust-based diet for the chameleons was determined. The diagnosis included the detailed description of clinical findings, histopathology, measurements of serum Ca, 25-hydroxycholecalciferol (25-OHD(3)), liver 25-OHD(3), vitamin A, bone mineral density, and bone mineral concentration. Chameleons that received no dietary supplementation of Ca, vitamin A, and cholecalciferol developed NMBD. When Ca and vitamin A were supplemented, the chameleons did not develop NMBD, independently of additional UVB and dietary cholecalciferol. The best prevention for NMBD was achieved by chameleons that received locusts gut-loaded with 12% Ca and dusted with 250,000 IU/kg (75 mg/kg) vitamin A and 25,000 IU/kg (0.625 mg/kg) cholecalciferol plus provision of long (10 h/d), low irradiation exposure (3-120 μW/cm(2)) to UVB. Chameleons that were fed diets low in vitamin A, cholecalciferol, and Ca were diagnosed with fibrous osteodystrophy. We noticed an interaction of vitamin A and cholecalciferol supplementation in the storage of vitamin A in the liver and formation of colon calcifications. From these findings, recommendations for the rearing of juvenile chameleons were derived.


Prevalence and determinants of unprotected sex in intimate partnerships of men who inject drugs: findings from a prospective intervention study.
Unprotected sex, common among people who inject drugs, puts them and their partners at risk of sexually transmitted infections including human immunodeficiency virus (HIV). This analysis assesses the changes in sexual risk behavior with regular female partners (RFPs), among married men who inject drugs, before and after implementation of a HIV prevention intervention, and identifies correlates of unprotected sex. People who inject drugs (PWID) were assessed at three points: baseline, preintervention follow-up visit (FV)1, and postintervention FV2. Descriptive analysis was used for reporting changes in sexual behavior over time. Generalized estimating equation assessed the population-averaged change in self-reported unprotected sex with an RFP, attributable to intervention uptake. Multivariable logistic regression determined correlates of self-reported unprotected sex with an RFP at FV2. Findings suggest that the proportion of men reporting any unprotected sex remained high (baseline = 46.0%, FV1 = 43.5%, FV2 = 37.0%). A reduction was observed in unprotected sex after the intervention phase, but this could not be attributed to uptake of the intervention. Higher odds of self-reported unprotected sex with an RFP in the past three months at FV2 were associated with self-reported unprotected sex at baseline, living with family, and being HIV-negative. Married male PWID should receive counseling for safe sex with RFPs, especially those who are HIV-negative and live with their families.


Accelerate Healing of Severe Burn Wounds by Mouse Bone Marrow Mesenchymal Stem Cell-Seeded Biodegradable Hydrogel Scaffold Synthesized from Arginine-Based Poly(ester amide) and Chitosan.
Severe burns are some of the most challenging problems in clinics and still lack ideal modalities. Mesenchymal stem cells (MSCs) incorporated with biomaterial coverage of burn wounds may offer a viable solution. In this report, we seeded MSCs to a biodegradable hybrid hydrogel, namely ACgel, that was synthesized from unsaturated arginine-based poly(ester amide) (UArg-PEA) and chitosan derivative. MSC adhered to ACgels. ACgels maintained a high viability of MSCs in culture for 6 days. MSC seeded to ACgels presented well in third-degree burn wounds of mice at 8 days postburn (dpb) after the necrotic full-thickness skin of burn wounds was debrided and filled and covered by MSC-carrying ACgels. MSC-seeded ACgels promoted the closure, reepithelialization, granulation tissue formation, and vascularization of the burn wounds. ACgels alone can also promote vascularization but less effectively compared with MSC-seeded ACgels. The actions of MSC-seeded ACgels or ACgels alone involve the induction of reparative, anti-inflammatory interleukin-10, and M2-like macrophages, as well as the reduction of inflammatory cytokine TNFα and M1-like macrophages at the late inflammatory phase of burn wound healing, which provided the mechanistic insights associated with inflammation and macrophages in burn wounds. For the studied regimens of these treatments, no toxicity was identified to MSCs or mice. Our results indicate that MSC-seeded ACgels have potential use as a novel adjuvant therapy for severe burns to complement commonly used skin grafting and, thus, minimize the downsides of grafting.


A simple yeast-based system for analyzing inhibitor resistance in the human cancer drug targets Hsp90alpha/beta.
Heat shock protein 90 (Hsp90), a highly conserved molecular chaperone, is one of the most promising targets for cancer drug development. Whether any resistance to these Hsp90 inhibitor drugs could arise by Hsp90 mutation is still unknown. Yeast is readily engineered so that its essential Hsp90 function is provided by either isoform of the human cytosolic Hsp90, Hsp90alpha or Hsp90beta. However, its high intrinsic resistance to most drugs poses a major obstacle to the use of such Hsp90alpha- or Hsp90beta-expressing yeast cells as a model system to analyse whether drug resistance might arise by Hsp90 mutation. In order to overcome this problem, we have generated a strain that is both hypersensitive to Hsp90 inhibitors as it lacks multiple drug resistance genes, and in which different heterologous and mutant Hsp90s can be expressed by plasmid exchange. It is not rendered appreciably stress sensitive when made to express Hsp90alpha or Hsp90beta as its sole form of Hsp90. Should there be any development of resistance to the Hsp90 drugs now in cancer clinic trials, this system can provide a rapid initial test of whether any single nucleotide polymorphism appearing within the coding regions of Hsp90alpha or Hsp90beta could be a contributory factor in this resistance. We have used this strain to demonstrate that significant levels of resistance to the Hsp90 inhibitors radicicol and 17-allylamino-demethoxygeldanamycin (17-AAG) are generated as a result of the same single point mutation within the native Hsp90 of yeast (A107N), the human Hsp90alpha (A121N) and the human Hsp90beta (A116N).


A Consensus Molecular Classification of Muscle-invasive Bladder Cancer.
Muscle-invasive bladder cancer (MIBC) is a molecularly diverse disease with heterogeneous clinical outcomes. Several molecular classifications have been proposed, but the diversity of their subtype sets impedes their clinical application. To achieve an international consensus on MIBC molecular subtypes that reconciles the published classification schemes. We used 1750 MIBC transcriptomic profiles from 16 published datasets and two additional cohorts. We performed a network-based analysis of six independent MIBC classification systems to identify a consensus set of molecular classes. Association with survival was assessed using multivariable Cox models. We report the results of an international effort to reach a consensus on MIBC molecular subtypes. We identified a consensus set of six molecular classes: luminal papillary (24%), luminal nonspecified (8%), luminal unstable (15%), stroma-rich (15%), basal/squamous (35%), and neuroendocrine-like (3%). These consensus classes differ regarding underlying oncogenic mechanisms, infiltration by immune and stromal cells, and histological and clinical characteristics, including outcomes. We provide a single-sample classifier that assigns a consensus class label to a tumor sample's transcriptome. Limitations of the work are retrospective clinical data collection and a lack of complete information regarding patient treatment. This consensus system offers a robust framework that will enable testing and validation of predictive biomarkers in future prospective clinical trials. Bladder cancers are heterogeneous at the molecular level, and scientists have proposed several classifications into sets of molecular classes. While these classifications may be useful to stratify patients for prognosis or response to treatment, a consensus classification would facilitate the clinical use of molecular classes. Conducted by multidisciplinary expert teams in the field, this study proposes such a consensus and provides a tool for applying the consensus classification in the clinical setting.


Targeting mitochondrial translation by inhibiting DDX3: a novel radiosensitization strategy for cancer treatment.
DDX3 is a DEAD box RNA helicase with oncogenic properties. RK-33 is developed as a small-molecule inhibitor of DDX3 and showed potent radiosensitizing activity in preclinical tumor models. This study aimed to assess DDX3 as a target in breast cancer and to elucidate how RK-33 exerts its anti-neoplastic effects. High DDX3 expression was present in 35% of breast cancer patient samples and correlated with markers of aggressiveness and shorter survival. With a quantitative proteomics approach, we identified proteins involved in the mitochondrial translation and respiratory electron transport pathways to be significantly downregulated after RK-33 or DDX3 knockdown. DDX3 localized to the mitochondria and DDX3 inhibition with RK-33 reduced mitochondrial translation. As a consequence, oxygen consumption rates and intracellular ATP concentrations decreased and reactive oxygen species (ROS) increased. RK-33 antagonized the increase in oxygen consumption and ATP production observed after exposure to ionizing radiation and reduced DNA repair. Overall, we conclude that DDX3 inhibition with RK-33 causes radiosensitization in breast cancer through inhibition of mitochondrial translation, which results in reduced oxidative phosphorylation capacity and increased ROS levels, culminating in a bioenergetic catastrophe.


Lacto-fermented sauerkraut improves symptoms in IBS patients independent of product pasteurisation - a pilot study.
Lacto-fermented sauerkraut contains a natural variety of lactic acid bacteria (LAB) and has not previously been studied in the treatment of irritable bowel syndrome (IBS) patients. The present study investigated the effect of a daily lacto-fermented sauerkraut supplement in relation to IBS patients' gastrointestinal symptoms and gut microbiota composition. A randomized double-blinded intervention was conducted with 34 Norwegian IBS patients. The patients were consuming either pasteurized sauerkraut (PS; n = 15) or unpasteurized sauerkraut (UPS; n = 19) as a supplement to their daily diet for 6 weeks. The differences in change of symptoms were assessed using the questionnaire IBS-Symptom Severity Score (IBS-SSS) measured at the baseline, and at weeks 2, 4, 6 and 8 (follow-up). The gut microbiota composition was analysed using 16S rRNA gene amplicon sequencing of faecal samples from the baseline and week 6. The mean change in IBS-SSS was -38.57 ± 17.08 PS vs. -56.99 ± 16.92 UPS and was significantly improved in both groups (P < 0.04), while the improvement in symptoms was not different between the intervention groups. The sauerkraut intervention (pasteurized or not) also led to significant gut microbiota compositional changes as determined by 16S rRNA gene amplicon sequencing (un-weighted UniFrac: P = 0.001, weighted UniFrac: P = 0.001). Sauerkraut related LAB in feces (Lactobacillus plantarum and Lactobacillus brevis) were significantly more often present in the UPS-group. In conclusion lacto-fermented sauerkraut had an effect on IBS patients' symptoms and gut microbiota even though the study was underpowered. Our results indicate that the observed effect to a larger extent can be attributed to the potential prebiotics in lacto-fermented sauerkraut rather than the viable LAB. Future studies with greater statistical power are needed to clarify the possible effects of LAB from lacto-fermented sauerkraut in the treatment of IBS patients.


Full Mitochondrial Genome and Nuclear 18S rDNA Sequences Refine the Taxonomic Placement of Choleoeimeria taggarti n. comb. from the Prostate of Antechinus flavipes (Yellow-Footed Antechinus).
An unusual coccidian parasite was described previously from the prostate of a male Antechinus flavipes (family: Dasyuridae; common name: yellow-footed antechinus). Morphometrics and a partial nuclear 18S small subunit rDNA (18S rDNA) sequence were used to assign this parasite to the genus Eimeria; it was named Eimeria taggarti. We generated full nuclear 18S rDNA and mitochondrial genome sequences from this parasite and used the newly completed 18S rDNA and mitochondrial cytochrome c oxidase subunit I (COI) sequences to perform a more in-depth phylogenetic analysis. The parasite clustered closely with Choleoeimeria spp. and Acroeimeria spp. infecting herptiles in a well-supported clade that was the sister lineage to the Eimeriidae sensu stricto. The mitochondrial genome of this parasite contained 2 inverted segments compared to mitochondrial genomes from parasites in the Eimeriidae sensu stricto (i.e., Stieda body-possessing coccidia with 4 dizoic sporocysts); this mitochondrial genome arrangement was shared with the only Choleoeimeria species for which sequence data were available publicly. Examination of histological preparations and TEM images uncovered bivalvate sporocysts and otherwise confirmed previously described morphological features of the parasite. Based on our phylogenetic analyses and histological observations, we propose the generic reclassification of E. taggarti to Choleoeimeria taggarti n. comb.


Strategy towards the enantioselective synthesis of schiglautone A.
Herein is described a convergent enantioselective route to an advanced intermediate in the synthesis of schiglautone A, a Schisandra triterpenoid with an unusual architecture. The synthetic route to this intermediate displaying 6 of the 7 stereocenters builds upon two fragments, an aldehyde elaborated from the Wieland-Miescher ketone, and a ketone. The preparation of the latter features a lithiation-borylation enzymatic resolution sequence, which led to the formation of the desired product with high enantio- and diastereoselectivities. After aldol coupling of the two fragments, the final quaternary stereocenter was installed by cyclopropane opening. The functionalized intermediate was isolated as a single diastereoisomer and thus offers a valuable starting point for further synthetic exploration.


Comparative X-ray standing wave analysis of metal-phosphonate multilayer films of dodecane and porphyrin molecular square.
The nanoscale structures of multilayer metal-phosphonate thin films prepared via a layer-by-layer assembly process using Zr(4+) and 1,12-dodecanediylbis(phosphonic acid) (DDBPA) or porphyrin square bis(phosphonic acid) (PSBPA) were studied using specular X-ray reflectivity (XRR), X-ray fluorescence, and long-period X-ray standing wave (XSW) analysis. The films were prepared in 1, 2, 3, 4, 6, and 8 layer series on both Si(001) substrates for XRR and on 18.6 nm period Si/Mo layered-synthetic microstructure X-ray mirrors for XSW. After functionalizing the SiO(2) substrate surfaces with a monolayer film terminated with phosphonate groups, the organic multilayer films were assembled by alternating immersions in (a) aqueous solutions containing Zr(4+)or Hf(4+) (final metal layer only) cations and then (b) organic solvent solutions of PO(3)-R-PO(3)(4-), where R was DDBPA or PSBPA spacer molecule. The Hf(4+) cation served as the marker for the top surface of the films, whereas the Zr(4+) cation was present in all other layers. The PSBPA also contained Zn and Re atoms at its midline which served as heavy-atom markers for each layer. The long-period XSW generated by the 0th- (total external reflection) through 4th-order Bragg diffraction conditions made it possible to examine the Fourier transforms of the fluorescent atom distributions over a much larger q(z) range in reciprocal space which permitted simultaneous analysis of Hf, Zn/Re, and Zr atomic distributions.


Detection of Chlamydia trachomatis in genital swabs: comparison of commercial and in house amplification methods with culture.
To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies (EB) and also to test 245 urethral and endocervical specimens for Chlamydia trachomatis by the four assays as well as conventional culture. EB titrations were run in duplicate in each commercial assay and six times in the in house PCR. Clinical samples were aliquoted and tested by each assay and were considered positive if C trachomatis was detected by two or more separate tests or if the sample was either culture or immunofluorescence positive. Major outer membrane protein (MOMP) specific primers were used as a confirmatory assay for the in house PCR. The in house PCR, Roche Cobas Amplicor, LCR, and Amplicor plate kit gave detection limits of approximately 1, 1-2, 2, and 2-4 EBs respectively. By the criteria described above for definition of a C trachomatis positive result in clinical samples we identified 23 true positives among the 245 clinical specimens. The in house PCR detected all 23 giving a sensitivity of 100% and a specificity of 98%. The Roche Cobas Amplicor, Roche Amplicor plate kit, and LCR detected 21, 19, and 19 of these respectively giving sensitivities of 87.5%, 82%, and 82% respectively and specificities of 99.5%, 99%, and 100% respectively. The culture gave a sensitivity of 78% and specificity of 100%. All four amplification assays had a greater sensitivity than the culture used routinely in this laboratory. The in house plasmid PCR had the greatest sensitivity and when combined with confirmation by immunofluorescence detected the greatest number of positives. This increased sensitivity is likely to have been achieved by the use of a DNA purification step and of nested primers in the amplification stage and their combined use in routine diagnostic assays for chlamydia might increase the frequency of C trachomatis detections. However, this assay is much less user friendly than the two semiautomated commercial assays investigated in this study.


Quasi-experimental trial of diabetes Self-Management Automated and Real-Time Telephonic Support (SMARTSteps) in a Medicaid managed care plan: study protocol.
Health information technology can enhance self-management and quality of life for patients with chronic disease and overcome healthcare barriers for patients with limited English proficiency. After a randomized controlled trial of a multilingual automated telephone self-management support program (ATSM) improved patient-centered dimensions of diabetes care in safety net clinics, we collaborated with a nonprofit Medicaid managed care plan to translate research into practice, offering ATSM as a covered benefit and augmenting ATSM to promote medication activation. This paper describes the protocol of the Self-Management Automated and Real-Time Telephonic Support Project (SMARTSteps). This controlled quasi-experimental trial used a wait-list variant of a stepped wedge design to enroll 362 adult health plan members with diabetes who speak English, Cantonese, or Spanish and receive care at 4 publicly-funded clinics. Through language-stratified randomization, participants were assigned to four intervention statuses: SMARTSteps-ONLY, SMARTSteps-PLUS, or wait-list for either intervention. In addition to usual primary care, intervention participants received 27 weekly calls in their preferred language with rotating queries and response-triggered education about self-care, medication adherence, safety concerns, psychological issues, and preventive services. Health coaches from the health plan called patients with out-of-range responses for collaborative goal setting and action planning. SMARTSteps-PLUS also included health coach calls to promote medication activation, adherence and intensification, if triggered by ATSM-reported non-adherence, refill non-adherence from pharmacy claims, or suboptimal cardiometabolic indicators. Wait-list patients crossed-over to SMARTSteps-ONLY or -PLUS at 6 months. For participants who agreed to structured telephone interviews at baseline and 6 months (n = 252), primary outcomes will be changes in quality of life and functional status with secondary outcomes of 6-month changes in self-management behaviors/efficacy and patient-centered processes of care. We will also evaluate 6-month changes in cardiometabolic (HbA1c, blood pressure, and LDL) and utilization indicators for all participants. Outcomes will provide evidence regarding real-world implementation of ATSM within a Medicaid managed care plan serving safety net settings. The evaluation trial will provide insight into translating and scaling up health information technology interventions for linguistically and culturally diverse vulnerable populations with chronic disease. ClinicalTrials.gov: NCT00683020.


Diagnosis of the hemolytic state using serum levels of erythrocyte adenylate kinase.
Red cell hemolysis is classically diagnosed by a combination of nonspecific laboratory tests, including serum bilirubin, LDH, and the reticulocyte count. None of these tests alone or in combination has the specificity to reliably ascertain the presence of hemolysis. We have previously demonstrated that erythrocyte adenylate kinase (EAK) is a red cell specific enzyme released from damaged red cells. Its activity can be measured in serum by rapid electrophoresis or immunological methods and correlates linearly with the degree of hemolysis in vitro. We now report on a clinical study comparing EAK levels in patients with and without hemolysis. The clinical diagnosis of hemolysis was established in hospitalized patients with anemia by the combined elevation of the bilirubin, LDH, and reticulocyte count in the absence of liver disease and demonstrable blood loss. The normal range of serum EAK was determined in 30 healthy nonanemic voluntary blood donors and was 0-3.5 Units (mean = 0.5). In 25 patients with hemolytic anemia due to sickle cell disease, hemolytic transfusion reactions, or TTP, the mean EAK level was 62.4 with a range 0-298 Units (P < 0.001 compared to normals). Levels of EAK exceeded the normal range in 24 of 25 patients (96%). In a control group of 44 hospitalized patients with liver disease or myocardial infarction and no clinical evidence of hemolysis, the mean EAK level was 0.12 with a range of 0-3.2 (P = 0.1, NS compared to normals and P < 0.001 compared to patients with hemolysis). None of the control patients had EAK levels that exceeded the normal range. The diagnostic sensitivity of the EAK assay for hemolysis, as calculated according to Baye's algorithm, was 96%, with a specificity and accuracy of 97%. Measurement of serum EAK represents a highly sensitive and specific test for the diagnosis of hemolytic anemia.


Physical Function Impairment and Frailty in Middle-Aged People Living With Human Immunodeficiency Virus in the REPRIEVE Trial Ancillary Study PREPARE.
People with human immunodeficiency virus (PWH) are at risk for accelerated development of physical function impairment and frailty; both associated with increased risk of falls, hospitalizations, and death. Identifying factors associated with physical function impairment and frailty can help target interventions. The REPRIEVE trial enrolled participants 40-75 years of age, receiving stable antiretroviral therapy with CD4+ T-cell count >100 cells/mm3, and with low to moderate cardiovascular disease risk. We conducted a cross-sectional analysis of those concurrently enrolled in the ancillary study PREPARE at enrollment. Among the 266 participants, the median age was 51 years; 81% were male, and 45% were black, and 28% had hypertension. Body mass index (BMI; calculated as weight in kilograms divided by height in meters squared) was 25 to <30 in 38% and ≥30 in 30%, 33% had a high waist circumference, 89% were physically inactive, 37% (95% confidence interval, 31%, 43%) had physical function impairment (Short Physical Performance Battery score ≤10), and 6% (4%, 9%) were frail and 42% prefrail. In the adjusted analyses, older age, black race, greater BMI, and physical inactivity were associated with physical function impairment; depression and hypertension were associated with frailty or prefrailty. Physical function impairment was common among middle-aged PWH; greater BMI and physical inactivity are important modifiable factors that may prevent further decline in physical function with aging. NCT02344290.


CP-25 inhibits PGE2-induced angiogenesis by down-regulating EP4/AC/cAMP/PKA-mediated GRK2 translocation.
G protein-coupled receptor kinase 2 (GRK2), a type of cytosolic enzyme, transiently translocates to the plasma membrane upon G protein-coupled receptors (GPCRs) activation, and it also binds to extracellular signal-regulated kinase (ERK) to inhibit the activation of ERK. GRK2 deficiency in endothelial cells (ECs) leads to increased pro-inflammatory signaling and promotes recruitment of leukocytes to activated ECs. However, the role of GRK2 in regulating angiogenesis remains unclear. Here, we show that GRK2 is a novel regulatory molecule on migration and tube formation of ECs, vessel sprouting ex vivo and angiogenesis in vivo. We identify that EP4/AC/cAMP/protein kinase A (PKA)-mediated GRK2 translocation to cells membrane decreases the binding of GRK2 and ERK1/2 to inhibit ERK1/2 activation, which promotes prostaglandin E2 (PGE2)-induced angiogenesis. GRK2 small interfering RNA (siRNA) inhibits the increase in PGE2-induced HUVECs migration and tube formation. In vivo, PGE2 increases ECs sprouting from normal murine aortic segments and angiogenesis in mice, but not from GRK2-deficient ones, on Matrigel. Further research found that Lys220 and Ser685 of GRK2 play an important role in angiogenesis by regulating GRK2 translocation. Paeoniflorin-6'-O-benzene sulfonate (CP-25), as a novel ester derivative of paeoniflorin (pae), has therapeutic potential for the treatment of adjuvant arthritis (AA) and collagen-induced arthritis (CIA), but the underlying mechanism of CP-25 on angiogenesis has not been elucidated. In our study, CP-25 inhibits the migration and tube formation of HUVECs, and angiogenesis in mice by down-regulating GRK2 translocation activation without affecting GRK2 total expression. Taken together, the present results revealed that CP-25 down-regulates EP4/AC/cAMP/PKA-mediated GRK2 translocation, restoring the inhibition of GRK2 for ERK1/2, thereby inhibiting PGE2-stimulated angiogenesis.


From hexamer to amyloid: marginal stability of apolipoprotein SAA2.2 leads to in vitro fibril formation at physiological temperature.
Serum amyloid A (SAA) is a major acute phase reactant and a small apolipoprotein of high density lipoproteins (HDL) in the serum. In cases of prolonged inflammation, SAA may form amyloid fibrils, leading to the disease of amyloid A (AA) amyloidosis. Recently, we have shown that murine SAA2.2, a non-amyloidogenic isoform in vivo, forms a hexamer in vitro containing a putative central channel. It is reported herein that upon thermal denaturation, hexameric SAA2.2 irreversibly dissociates to a misfolded monomer at physiological temperature, formation of which coincides with a significant loss of alpha-helical and gain of beta-sheet structure. When SAA2.2 is incubated for several days at 37 degrees C, sedimentation analytical ultracentrifugation reveals the presence of soluble high molecular weight aggregates, which upon further incubation undergo subsequent self-assembly into amyloid fibrils. Limited proteolysis experiments suggest that the in vitro amyloidogenecity of SAA2.2 is related to structural alteration in its N-terminus. Our observation that SAA2.2 can form amyloid fibrils in vitro at physiological temperatures suggests that SAA2.2's inability to cause amyloidosis may be related to other factors, such as the stabilization of hexameric SAA2.2 (possibly through ligand binding), and/or the slow kinetics of aberrant misfolding and self-assembly.


The use of starter cultures in the table olive fermentation can modulate the antiadhesive properties of brine exopolysaccharides against enterotoxigenic Escherichia coli.
The present study aimed to evaluate different mates of Candida boidinii and Lactobacillus pentosus strains as starters in green table olive fermentation. Changes in fermentation characteristics as well as changes in the functional properties of the microbial exopolysaccharides (EPS) produced during the process were registered. The in vitro adhesion test demonstrated that most EPS samples could specifically attach ETEC K88. In vitro studies with porcine intestinal cells showed the improved blocking activity of the fimbria (blocking test) when the mutant strain L. pentosus 119-14MT was used alone as a starter. All EPS samples showed the ability to block receptors in the cells (exclusion test) although without differences between starter treatments. In the displacement test, EPS samples failed to remove the pathogen once attached. According to these results, L. pentosus 119-14MT, a high EPS variant, seemed to be the most effective starter improving the anti-adhesive properties of brine EPS and increasing its ability to block the ETEC K88 fimbria. These results illustrate that the anti-adhesive properties of the EPSs produced during the traditional fermentation of olives could be modulated by the use of defined starters. This opens the door to new fermentation processes aimed to produce green table olives as functional food to prevent ETEC diarrhea.


Structural mechanism of glutamate receptor activation and desensitization.
Ionotropic glutamate receptors are ligand-gated ion channels that mediate excitatory synaptic transmission in the vertebrate brain. To gain a better understanding of how structural changes gate ion flux across the membrane, we trapped rat AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid) and kainate receptor subtypes in their major functional states and analysed the resulting structures using cryo-electron microscopy. We show that transition to the active state involves a 'corkscrew' motion of the receptor assembly, driven by closure of the ligand-binding domain. Desensitization is accompanied by disruption of the amino-terminal domain tetramer in AMPA, but not kainate, receptors with a two-fold to four-fold symmetry transition in the ligand-binding domains in both subtypes. The 7.6 Å structure of a desensitized kainate receptor shows how these changes accommodate channel closing. These findings integrate previous physiological, biochemical and structural analyses of glutamate receptors and provide a molecular explanation for key steps in receptor gating.


Therapeutic potential of aryl hydrocarbon receptor in autoimmunity.
Aryl hydrocarbon receptor (AhR), a type of transcriptional factor, is widely expressed in immune cells. The activation of AhR signaling pathway depends on its ligands, which exist in environment and can also be produced by metabolism. Normal expressions of AhR and AhR-mediated signaling may be essential for immune responses, and effects of AhR signaling on the development and function of innate and adaptive immune cells have also been revealed in previous studies. Recent studies also indicate that aberrant AhR signaling may be related to autoimmune diseases, including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), multiple sclerosis (MS), autoimmune uveitis (AU), autoimmune diabetes, Behcet's disease (BD) and myasthenia gravis (MG). Moreover, administration of AhR ligands or drugs has been proven effective for improving pathological outcomes in some autoimmune diseases or models. In this review, we summarize the effects of AhR on several innate and adaptive immune cells associated with autoimmunity, and the mechanism on how AhR participates in autoimmune diseases. In addition, we also discuss therapeutic potential and application prospect of AhR in autoimmune diseases, so as to provide valuable information for exploring novel and effective approaches to autoimmune disease treatments.


Carrier and thermal dynamics of silicon photonic resonators at cryogenic temperatures.
We describe measurement results of silicon photonic circuits at cryogenic temperatures. The interplay between optically induced heating and free carrier dynamics in nano-photonic ring resonators is investigated at temperatures down to 1.8 K. We find that the life-time of free carriers generated by two-photon absorption in silicon waveguides is reduced from 1.9 ns at room temperature to less than 100 ps below 10K. At the same time the thermal relaxation time is significantly elongated. Our work provides the first cryogenic measurement of ultra-short free-carrier lifetimes in silicon waveguides. The results further indicate that integrated optical chips can be easily thermo-optically stabilized at low temperatures.


Resource capture and competitive ability of non-pathogenic Pseudogymnoascus spp. and P. destructans, the cause of white-nose syndrome in bats.
White-nose syndrome (WNS) is a devastating fungal disease that has been causing the mass mortality of hibernating bats in North America since 2006 and is caused by the psychrophilic dermatophyte Pseudogymnoascus destructans. Infected bats shed conidia into hibernaculum sediments and surfaces, but it is unknown if P. destructans can form stable, reproductive populations outside its bat hosts. Previous studies have found non-pathogenic Pseudogymnoascus in bat hibernacula, and these fungi may provide insight into the natural history of P. destructans. We compared the relatedness, resource capture, and competitive ability of non-pathogenic Pseudogymnoascus isolates with P. destructans to determine if they have similar adaptations for survival in hibernacula sediment. All non-pathogenic Pseudogymnoascus isolates grew faster, utilized a broader range of substrates with higher efficiency, and were generally more resistant to antifungals compared to P. destructans. All isolates also showed the ability to displace P. destructans in co-culture assays, but only some produced extractible antifungal metabolites. These results suggest that P. destructans would perform poorly in the same environmental niche as non-pathogenic Pseudogymnoascus, and must have an alternative saprophytic survival strategy if it establishes active populations in hibernaculum sediment and non-host surfaces.


Disparate phylogeographic patterns of molecular genetic variation in four closely related South American small cat species.
Tissue specimens from four species of Neotropical small cats (Oncifelis geoffroyi, N = 38; O. guigna, N = 6; Leopardus tigrinus, N = 32; Lynchailurus colocolo, N = 22) collected from throughout their distribution were examined for patterns of DNA sequence variation using three mitochondrial genes, 16S rRNA, ATP8, and NADH-5. Patterns between and among O. guigna and O. geoffroyi individuals were assessed further from size variation at 20 microsatellite loci. Phylogenetic analyses using mitochondrial DNA sequences revealed monophyletic clustering of the four species, plus evidence of natural hybridization between L. tigrinus and L. colocolo in areas of range overlap and discrete population subdivisions reflecting geographical isolation. Several commonly accepted subspecies partitions were affirmed for L. colocolo, but not for O. geoffroyi. The lack of geographical substructure in O. geoffroyi was recapitulated with the microsatellite data, as was the monophyletic clustering of O. guigna and O. geoffroyi individuals. L. tigrinus forms two phylogeographic clusters which correspond to L.t. oncilla (from Costa Rica) and L.t. guttula (from Brazil) and which have mitochondrial DNA (mtDNA) genetic distance estimates comparable to interspecific values between other ocelot lineage species. Using feline-specific calibration rates for mitochondrial DNA mutation rates, we estimated that extant lineages of O. guigna diverged 0.4 million years ago (Ma), compared with 1.7 Ma for L. colocolo, 2.0 Ma for O. geoffroyi, and 3.7 Ma for L. tigrinus.


Effect of Abdominal Loading Location on Liver Motion: Experimental Assessment using Ultrafast Ultrasound Imaging and Simulation with a Human Body Model.
A protocol based on ultrafast ultrasound imaging was applied to study the in situ motion of the liver while the abdomen was subjected to compressive loading at 3 m/s by a hemispherical impactor or a seatbelt. The loading was applied to various locations between the lower abdomen and the mid thorax while feature points inside the liver were followed on the ultrasound movie (2000 frames per second). Based on tests performed on five post mortem human surrogates (including four tested in the current study), trends were found between the loading location and feature point trajectory parameters such as the initial angle of motion or the peak displacement in the direction of impact. The impactor tests were then simulated using the GHBMC M50 human body model that was globally scaled to the dimensions of each surrogate. Some of the experimental trends observed could be reproduced in the simulations (e.g. initial angle) while others differed more widely (e.g. final caudal motion). The causes for the discrepancies need to be further investigated. The liver strain energy density predicted by the model was also widely affected by the impact location. Experimental and simulation results both highlight the importance of the liver position with respect to the impactor when studying its response in situ.


The 2018 Revised FDA Guidance for Early Alzheimer's Disease: Establishing the Meaningfulness of Treatment Effects.
The present report reviews the revised 2018 FDA guidance for early AD, with an emphasis on meaningfulness of clinical outcome assessments (COAs). A radical shift is evident in the importance given to establishing the meaningfulness of COAs in the 2018 draft versus the 2013 draft. The implications of this shift include the assertion that cognition is clinically meaningful, but that a persuasive effect on cognition, depending upon disease stage of the participants in the trial, is one that is of enough magnitude, established across multiple relevant domains, and can be supported by biomarkers reflecting underlying AD pathological changes. Meaningfulness is established through an understanding of the conceptual relevance of what is being measured and magnitude of any treatment effect. Precedent exists within other FDA guidance and independent good practices publications as to how meaningfulness may be assessed e.g. via evaluation of content validity and concepts such as minimally important difference. Additionally, FDA is developing a series of methodological Patient Focused Drug Development (PFDD) documents to provide further guidance on this topic, which are aimed at addressing gaps in methodology and recommended best practice. Importantly, application of PFDD approaches to AD is behind that in other areas and there is limited published content validity for COAs and a lack of supportive qualitative research. Initiatives to build robust conceptual models of AD and develop novel direct measures of meaningful health outcomes will have a significant impact on measurement of efficacy in clinical trials and on payer determinations of beneficiary value. Greater recognition of what is meaningful from the perspective of the patient and caregiver will inform regulatory reviews and determinations for payment and coverage of treatments.


Nutrient-based therapies for bipolar disorder: a systematic review.
Pharmacotherapy is the first line of treatment for bipolar disorder, but many patients continue to experience persistent subthreshold symptoms. Alternative adjunct treatments, including nutritional therapies, may have the potential to alleviate residual symptoms and improve the outcomes of standard pharmacotherapy. The aim of this paper is to critically review the current clinical evidence and mechanisms of action of nutrient-based therapies alone or in combination with commonly used pharmacotherapies for mania and bipolar depression. We conducted a Medline search for clinical trials conducted with humans, published in English from 1960 to 2012 using nutritional supplements such as n-3, chromium, inositol, choline, magnesium, folate and tryptophan alone or in combination with pharmacotherapies for the treatment of bipolar disorder. Preliminary data yields conflicting but mainly positive evidence for the use of n-3 fatty acids and chromium in the treatment of bipolar depression. Limited evidence found that inositol may be helpful for bipolar depression, but larger sample sizes are needed. Preliminary randomized, controlled trials suggest that choline, magnesium, folate and tryptophan may be beneficial for reducing symptoms of mania. Given the potential public health impact of identifying adjunct treatments that improve psychiatric as well as physical health outcomes, nutritional treatments appear promising for the management of bipolar disorder but require further study.


Retinal haemangioblastomas in von Hippel-Lindau germline mutation carriers: progression, complications and treatment outcome.
Evaluation of phenotype and treatment outcome of retinal haemangioblastomas (RH) in von Hippel-Lindau (VHL) disease and correlation of these features with the genotype of VHL germline mutation carriers. Retrospective analysis of a longitudinal cohort of 21 VHL germline mutation carriers and RH. Clinical and genetic data were obtained to analyse the correlation of genotype with phenotype and treatment outcomes. All patients were categorized in two genotypic categories: missense mutations (MM) and truncating mutations (TM). Mean follow-up duration was 16.3 years and did not differ significantly between mutation groups (p = 0.383). Missense mutations (MM) carriers (n = 6) developed more progression-related complications compared to TM carriers (n = 15) (p = 0.046). Vitreoretinal surgery was more often applied in MM carriers (p = 0.036). Moderate (visual acuity (VA)20/80 to 20/200) to severe (VA < 20/200) visual impairment was observed in 53.3% of the eyes of MM carriers and 28.1% of the eyes of TM carriers at last recorded visit. Missense mutations in VHL patients seem to have a higher prevalence of progression-related complications. Missense mutations (MM) carriers required therefore more often vitreoretinal surgical treatment with a worse treatment outcome. Genetic analysis may play a role in determining a pro-active treatment strategy and prognosis for RH.


Mechanisms of ascorbate-induced cytotoxicity in pancreatic cancer.
Pharmacologic concentrations of ascorbate may be effective in cancer therapeutics. We hypothesized that ascorbate concentrations achievable with i.v. dosing would be cytotoxic in pancreatic cancer for which the 5-year survival is <3%. Pancreatic cancer cell lines were treated with ascorbate (0, 5, or 10 mmol/L) for 1 hour, then viability and clonogenic survival were determined. Pancreatic tumor cells were delivered s.c. into the flank region of nude mice and allowed to grow at which time they were randomized to receive either ascorbate (4 g/kg) or osmotically equivalent saline (1 mol/L) i.p. for 2 weeks. There was a time- and dose-dependent increase in measured H(2)O(2) production with increased concentrations of ascorbate. Ascorbate decreased viability in all pancreatic cancer cell lines but had no effect on an immortalized pancreatic ductal epithelial cell line. Ascorbate decreased clonogenic survival of the pancreatic cancer cell lines, which was reversed by treatment of cells with scavengers of H(2)O(2). Treatment with ascorbate induced a caspase-independent cell death that was associated with autophagy. In vivo, treatment with ascorbate inhibited tumor growth and prolonged survival. These results show that pharmacologic doses of ascorbate, easily achievable in humans, may have potential for therapy in pancreatic cancer.


Alternative uses for co-products: Harnessing the potential of valuable compounds from meat processing chains.
Opportunities for exploiting the inherent value of protein-rich meat processing co-products, in the context of increased global demand for protein and for sustainable processing systems, are discussed. While direct consumption maybe the most profitable route for some, this approach is influenced greatly by local and cultural traditions. A more profitable and sustainable approach may be found in recognizing this readily available and under-utilised resource can provide high value components, such as proteins, with targeted high value functionality of relevance to a variety of sectors. Applications in food & beverages, petfood biomedical and nutrition arenas are discussed. Utilization of the raw material in its entirety is a necessary underlying principle in this approach to help maintain minimum waste generation. Understanding consumer attitudes to these products, in particular when used in food or beverage systems, is critical in optimizing commercialization strategies.


The role of hypoxia-inducible factor-1α in zinc oxide nanoparticle-induced nephrotoxicity in vitro and in vivo.
Zinc oxide nanoparticles (ZnO NPs) are used in an increasing number of products, including rubber manufacture, cosmetics, pigments, food additives, medicine, chemical fibers and electronics. However, the molecular mechanisms underlying ZnO NP nephrotoxicity remain unclear. In this study, we evaluated the potential toxicity of ZnO NPs in kidney cells in vitro and in vivo. We found that ZnO NPs were apparently engulfed by the HEK-293 human embryonic kidney cells and then induced reactive oxygen species (ROS) generation. Furthermore, exposure to ZnO NPs led to a reduction in cell viability and induction of apoptosis and autophagy. Interestingly, the ROS-induced hypoxia-inducible factor-1α (HIF-1α) signaling pathway was significantly increased following ZnO NPs exposure. Additionally, connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1), which are directly regulated by HIF-1 and are involved in the pathogenesis of kidney diseases, displayed significantly increased levels following ZnO NPs exposure in HEK-293 cells. HIF-1α knockdown resulted in significantly decreased levels of autophagy and increased cytotoxicity. Therefore, our results suggest that HIF-1α may have a protective role in adaptation to the toxicity of ZnO NPs in kidney cells. In an animal study, fluorescent ZnO NPs were clearly observed in the liver, lungs, kidneys, spleen and heart. ZnO NPs caused histopathological lesions in the kidney and increase in serum creatinine and blood urea nitrogen (BUN) which indicate possible renal possible damage. Moreover, ZnO NPs enhanced the HIF-1α signaling pathway, apoptosis and autophagy in mouse kidney tissues. ZnO NPs may cause nephrotoxicity, and the results demonstrate the importance of considering the toxicological hazards of ZnO NP production and application, especially for medicinal use.


The rate of device-associated nosocomial infections in a medical surgical intensive care unit of a training and research hospital in Turkey: one-year outcomes.
In the present study, we aimed to assess the rate and effect of device-associated nosocomial infections (DANIs), as well as the rate of antibiotic resistance, in the medical-surgical intensive care unit (ICU) of a research and training hospital in Turkey, and to compare our results with those reported by the National Nosocomial Infections Surveillance (NNIS) system and International Nosocomial Infection Control Consortium (INICC). A total of 509 patients were followed up within a 1-year period from 1 November 2007 to 1 November 2008. The total patient days were 4,087, the number of DANIs was 181. The ventilator-associated pneumonia rate in 1,000 ventilator days was 27.1, the rate of central venous catheter (CVC)-associated blood circulation infections in 1,000 CVC days was 11.8, and the rate of urinary catheter-associated urinary tract infections in 1,000 urinary catheter days was 9.6. The most frequently isolated microorganisms were Pseudomonas aeruginosa and Acinetobacter spp. Of the infections caused by Staphylococcus aureus, 81.2% were due to methicillin-resistant strains. Of the Enterobacteriaceae isolates, 53.5% were found to be resistant to ceftriaxone, while 29% of the P. aeruginosa isolates were found to be resistant to ciprofloxacin. The rates of use of devices such as ventilators, CVCs, and urinary catheters were 0.87, 0.93, and 0.98, respectively, which are higher than the rates reported by NNIS and INICC. On the other hand, the present DANI rate was higher than that reported by NNIS, but close to that reported by INICC. We conclude that the indications for and duration of device use should be reviewed.


Neuroscience20 (BRAIN20, SPINE20, and MENTAL20) Health Initiative: A Global Consortium Addressing the Human and Economic Burden of Brain, Spine, and Mental Disorders Through Neurotech Innovations and Policies.
Neurological disorders significantly impact the world's economy due to their often chronic and life-threatening nature afflicting individuals which, in turn, creates a global disease burden. The Group of Twenty (G20) member nations, which represent the largest economies globally, should come together to formulate a plan on how to overcome this burden. The Neuroscience-20 (N20) initiative of the Society for Brain Mapping and Therapeutics (SBMT) is at the vanguard of this global collaboration to comprehensively raise awareness about brain, spine, and mental disorders worldwide. This paper aims to provide a comprehensive review of the various brain initiatives worldwide and highlight the need for cooperation and recommend ways to bring down costs associated with the discovery and treatment of neurological disorders. Our systematic search revealed that the cost of neurological and psychiatric disorders to the world economy by 2030 is roughly $16T. The cost to the economy of the United States is $1.5T annually and growing given the impact of COVID-19. We also discovered there is a shortfall of effective collaboration between nations and a lack of resources in developing countries. Current statistical analyses on the cost of neurological disorders to the world economy strongly suggest that there is a great need for investment in neurotechnology and innovation or fast-tracking therapeutics and diagnostics to curb these costs. During the current COVID-19 pandemic, SBMT, through this paper, intends to showcase the importance of worldwide collaborations to reduce the population's economic and health burden, specifically regarding neurological/brain, spine, and mental disorders.