Contribution of Complex I NADH Dehydrogenase to Respiratory Energy Coupling in Glucose-Grown Cultures of Ogataea parapolymorpha.
The thermotolerant yeast Ogataea parapolymorpha (formerly Hansenula polymorpha) is an industrially relevant production host that exhibits a fully respiratory sugar metabolism in aerobic batch cultures. NADH-derived electrons can enter its mitochondrial respiratory chain either via a proton-translocating complex I NADH-dehydrogenase or via three putative alternative NADH dehydrogenases. This respiratory entry point affects the amount of ATP produced per NADH/O2 consumed and therefore impacts the maximum yield of biomass and/or cellular products from a given amount of substrate. To investigate the physiological importance of complex I, a wild-type O. parapolymorpha strain and a congenic complex I-deficient mutant were grown on glucose in aerobic batch, chemostat, and retentostat cultures in bioreactors. In batch cultures, the two strains exhibited a fully respiratory metabolism and showed the same growth rates and biomass yields, indicating that, under these conditions, the contribution of NADH oxidation via complex I was negligible. Both strains also exhibited a respiratory metabolism in glucose-limited chemostat cultures, but the complex I-deficient mutant showed considerably reduced biomass yields on substrate and oxygen, consistent with a lower efficiency of respiratory energy coupling. In glucose-limited retentostat cultures at specific growth rates down to ∼0.001 h-1, both O. parapolymorpha strains showed high viability. Maintenance energy requirements at these extremely low growth rates were approximately 3-fold lower than estimated from faster-growing chemostat cultures, indicating a stringent-response-like behavior. Quantitative transcriptome and proteome analyses indicated condition-dependent expression patterns of complex I subunits and of alternative NADH dehydrogenases that were consistent with physiological observations.IMPORTANCE Since popular microbial cell factories have typically not been selected for efficient respiratory energy coupling, their ATP yields from sugar catabolism are often suboptimal. In aerobic industrial processes, suboptimal energy coupling results in reduced product yields on sugar, increased process costs for oxygen transfer, and volumetric productivity limitations due to limitations in gas transfer and cooling. This study provides insights into the contribution of mechanisms of respiratory energy coupling in the yeast cell factory Ogataea parapolymorpha under different growth conditions and provides a basis for rational improvement of energy coupling in yeast cell factories. Analysis of energy metabolism of O. parapolymorpha at extremely low specific growth rates indicated that this yeast reduces its energy requirements for cellular maintenance under extreme energy limitation. Exploration of the mechanisms for this increased energetic efficiency may contribute to an optimization of the performance of industrial processes with slow-growing eukaryotic cell factories.


The role of putative fibrinogen Aalpha-, Bbeta-, and GammaA-chain integrin binding sites in endothelial cell-mediated clot retraction.
In this study, endothelial cell-mediated clot retraction was supported by fibrin generated from several purified fractions of plasma fibrinogen, purified proteolytic fragments of plasma fibrinogen, recombinant normal fibrinogen, and recombinant variant fibrinogen. These results were surprising because some of these fibrinogens lack domains that are known binding sites for the integrin receptors that support clot retraction. Specifically, fibrinogens lacking Aalpha-chain RGD residues at 572-574 or lacking the gamma-chain residues AGDV 408-411 supported endothelial cell-mediated clot retraction as well as intact fibrinogen. Thus, clot retraction mediated by endothelial cells is not dependent on either of these sites. A variety of monoclonal antibodies against the integrin alphavbeta3 partially inhibited the endothelial cell-mediated retraction of clots formed from plasma fibrinogen. As expected, an antibody to the platelet integrin alphaIIbbeta3 did not inhibit endothelial cell-mediated clot retraction. These results indicate that this retraction is mediated at least in part by alphavbeta3. These results support the conclusion that (a) neither of the two fibrinogen cell binding sites described above is required to support clot retraction or that (b) either site alone or in conjunction with other fibrin(ogen) region(s) can support clot retraction. Thus, endothelial cell-mediated clot retraction appears to be dependent on fibrinogen cell binding sites other than those required to support adhesion of resting platelets to immobilized fibrinogen and platelet aggregation.


Assessment of Nine Driver Gene Mutations in Surgically Resected Samples from Patients with Non-Small-Cell Lung Cancer.
The mutational profile of oncogenic driver genes play an important role in non-small-cell lung cancer (NSCLC). The need of a testing panel capable of comprehensively determining patient genotypes in limited amounts of material has increased since the recent association of nine core oncogenic driver genes as tumor predictive biomarkers. Surgically resected samples from 214 NSCLC patients (168 patients with adenocarcinomas and 46 with squamous cell cancers) were included. A multiplexed PCR-based assay was developed to simultaneously test 118 hotspot mutations and fusions in nine driver genes. The sensitivity of the kit was 1% for gene mutation and 450 copies for gene fusion. Genetic alterations were detected in 143 (66.8%) patients by the assay. The three most common alterations identified were EGFR mutations (50.9%), KRAS mutations (8.4%) and ALK fusions (4.7%). Eight (3.7%) patients harbored concurrent mutations, and the most common partners were EGFR mutations which were observed in the eight patients. No associations between survival and EGFR, KRAS, and ALK status were observed. Patients with two or more alterations exhibited shorter DFS compared to those with single mutations (P=0.032), whilst had no significant difference in OS (P=0.245). However, only TNM stage was an independent predictor of OS (HR=2.905, P<0.001) as well as DFS (HR=2.114, P<0.001) in our cohort in multivariate analysis. Furthermore, patients with the L858R mutation had longer DFS (P=0.014) compared to other sensitizing mutations and tended to have better OS but the differences were not significant (P=0.06). These findings suggest this multiplex gene panel testing technique can be efficiently used to detect nine driver genes in a limited number of specimens. This methodology would have the potential to save both specimens and time compared to the combination of all assays by other methods.


Motor Neuron Diseases Accompanying Spinal Stenosis: A Case Study.
A 75-year-old man, who was healthy, visited the hospital because of shooting pain and numbness in both lower limbs (right > left). The patient had an L4/5 moderate right foraminal stenosis and right subarticular disc protrusion and received a lumbar epidural block. The patient experienced severe weakness in the right lower limb after 2 days. Lumbar and cervical magnetic resonance images were taken and electromyography and a nerve conduction study were performed to arrive at the diagnosis of a motor neuron disease. The patient expired 4 months later with respiratory failure due to motor neuron disease. This case suggests that any abnormal neurological symptoms that occur after an epidural block should be examined thoroughly via testing and consultations to identify the cause of the symptoms.


The determination of manganese in urine.
A simple and quite sensitive methods of manganese determination in urine is proposed. The manganese level as well as the fluoride level are directly proportional to the level of these substances in plasma and it doesn't depend substantially on the diuresis. In not exposed population the manganese level of 24 +/- 4 nmol.l-1 (mean +/- +/- 2 SEM) was found, in population exposed to the manganese containing aerosole as much as 3500 nmol.l-1 without any signs of health damage. The level of 400 nmol.l-1 is suggested to be a safe biological limit. During the shift, the manganese level in urine evidently raises. The simultaneous inhalation of manganese and fluoride results in their interaction.


Soft tissue grafting to improve the attached mucosa at dental implants: A review of the literature and proposal of a decision tree.
Scientific data and clinical observations appear to indicate that an adequate width of attached mucosa may facilitate oral hygiene procedures thus preventing peri-implant inflammation and tissue breakdown (eg, biologic complications). Consequently, in order to avoid biologic complications and improve long-term prognosis, soft tissue conditions should be carefully evaluated when implant therapy is planned. At present the necessity and time-point for soft tissue grafting (eg, prior to or during implant placement or after healing) is still controversially discussed while clinical recommendations are vague. To provide a review of the literature on the role of attached mucosa to maintain periimplant health, and to propose a decision tree which may help the clinician to select the appropriate surgical technique for increasing the width of attached mucosa. The available data indicate that ideally, soft tissue conditions should be optimized by various grafting procedures either before or during implant placement or as part of stage-two surgery. In cases, where, despite insufficient peri-implant soft tissue condition (ie, lack of attached mucosa or movements caused by buccal frena), implants have been uncovered and/or loaded, or in cases where biologic complications are already present (eg, mucositis, peri-implantitis), the treatment appears to be more difficult and less predictable. Soft tissue grafting may be important to prevent peri-implant tissue breakdown and should be considered when dental implants are placed. The presented decision tree may help the clinician to select the appropriate grafting technique.


Inferring the Chemotactic Strategy of P. putida and E. coli Using Modified Kramers-Moyal Coefficients.
Many bacteria perform a run-and-tumble random walk to explore their surrounding and to perform chemotaxis. In this article we present a novel method to infer the relevant parameters of bacterial motion from experimental trajectories including the tumbling events. We introduce a stochastic model for the orientation angle, where a shot-noise process initiates tumbles, and analytically calculate conditional moments, reminiscent of Kramers-Moyal coefficients. Matching them with the moments calculated from experimental trajectories of the bacteria E. coli and Pseudomonas putida, we are able to infer their respective tumble rates, the rotational diffusion constants, and the distributions of tumble angles in good agreement with results from conventional tumble recognizers. We also define a novel tumble recognizer, which explicitly quantifies the error in recognizing tumbles. In the presence of a chemical gradient we condition the moments on the bacterial direction of motion and thereby explore the chemotaxis strategy. For both bacteria we recover and quantify the classical chemotactic strategy, where the tumble rate is smallest along the chemical gradient. In addition, for E. coli we detect some cells, which bias their mean tumble angle towards smaller values. Our findings are supported by a scaling analysis of appropriate ratios of conditional moments, which are directly calculated from experimental data.


Outreach to diversify clinical trial participation: A randomized recruitment study.
Racial and ethnic minorities remain underrepresented in clinical research, yet few recruitment strategies have been rigorously evaluated. We experimentally tested whether targeted recruitment letters acknowledging diabetes health disparities and health risks specific to recipients' racial/ethnic group improved two metrics of trial participation: willingness to be screened and enrollment. This experiment was efficiently nested within a randomized clinical trial examining a preventive lifestyle intervention among women at high risk for diabetes. Pregnant women with gestational diabetes or impaired glucose tolerance (N = 445) were randomized to receive a targeted recruitment letter with health risk information specific to their racial/ethnic group (n = 216), or a standard letter with risk information for the general population (n = 229). All letters were bilingual in English and Spanish. The targeted as compared to the standard letter did not improve screening or enrollment rates overall or within separate racial/ethnic groups. Among Latina women who preferred Spanish, the targeted letter showed trends for improved screening (66.7% vs 33.3%, p = .06) and enrollment rates (38.9% vs 13.3%, p = .13). In contrast, among Latina women who preferred English, the targeted letter significantly lowered screening (29.6% vs 57.1%, p = .04) and showed trends for lowered enrollment rates (25.9% vs 50.0%, p = .07). Results from this randomized study appear to suggest that recruitment letters with diabetes health risk information targeted to recipients' race/ethnicity may improve one metric of clinical trial participation among Latina women who prefer Spanish, but not English. Larger experimental studies, incorporating input from diverse participant stakeholders, are needed to develop evidence-based minority recruitment strategies.


Subpolar link to the emergence of the modern equatorial Pacific cold tongue.
The cold upwelling "tongue" of the eastern equatorial Pacific is a central energetic feature of the ocean, dominating both the mean state and temporal variability of climate in the tropics and beyond. Recent evidence for the development of the modern cold tongue during the Pliocene-Pleistocene transition has been explained as the result of extratropical cooling that drove a shoaling of the thermocline. We have found that the sub-Antarctic and sub-Arctic regions underwent substantial cooling nearly synchronous to the cold tongue development, thereby providing support for this hypothesis. In addition, we show that sub-Antarctic climate changed in its response to Earth's orbital variations, from a subtropical to a subpolar pattern, as expected if cooling shrank the warm-water sphere of the ocean and thus contracted the subtropical gyres.


Effect of iron-, iodine-, and beta-carotene-fortified biscuits on the micronutrient status of primary school children: a randomized controlled trial.
Deficiencies of iron, iodine, and vitamin A are prevalent worldwide and can affect the mental development and learning ability of schoolchildren. The aim of this study was to determine the effect of micronutrient-fortified biscuits on the micronutrient status of primary school children. Micronutrient status was assessed in 115 children aged 6-11 y before and after consumption of biscuits (fortified with iron, iodine, and beta-carotene) for 43 wk over a 12-mo period and was compared with that in a control group (n = 113) who consumed nonfortified biscuits. Cognitive function, growth, and morbidity were assessed as secondary outcomes. There was a significant between-group treatment effect on serum retinol, serum ferritin, serum iron, transferrin saturation, and urinary iodine (P <0.0001) and in hemoglobin and hematocrit (P <0.05). The prevalence of low serum retinol concentrations (<0.70 micromol/L) decreased from 39.1% to 12.2%, of low serum ferritin concentrations (<20 microg/L) from 27.8% to 13.9%, of anemia (hemoglobin <120 g/L) from 29.6% to 15.6%, and of low urinary iodine concentrations (<100 microg/L) from 97.5% to 5.4%. There was a significant between-group treatment effect (P <0.05) in cognitive function with the digit span forward task (short-term memory). Fewer school days were missed in the intervention than in the control group because of respiratory- (P = 0.097) and diarrhea-related (P = 0.013) illnesses. The intervention had no effect on anthropometric status [corrected]. Fortified biscuits resulted in a significant improvement in the micronutrient status of primary school children from a poor rural community and also appeared to have a favorable effect on morbidity and cognitive function [corrected].


Haptoglobin phenotype prevalence and cytokine profiles during Plasmodium falciparum infection in Dogon and Fulani ethnic groups living in Mali.
The Fulani are known to have a lower parasitaemia and less clinical episodes of malaria as compared to the Dogon sympatric ethnic group, living in Mali. Higher circulating malaria-specific antibody titers and increased pro-inflammatory cytokine levels have been shown in Fulani individuals. Several studies have tried to link haptoglobin (Hp) phenotypes with susceptibility to malaria, but without consensus. This study investigated the role of Hp phenotypes and cytokine levels in Dogon and Fulani during asymptomatic Plasmodium falciparum infection. Two different cohorts were combined in this study: a 2008 cohort with 77 children aged between two and ten years and a 2001 cohort, with 82 children and adults, aged between 11 and 68 years. Hp phenotypes in plasma were measured by Western Blot. Circulating levels of sCD163, IL-6, IL-10, IFN-γ and TNF were measured by ELISA. Multiple regression analysis was performed to associate Hp phenotypes with cytokine profiles. In addition, in vitro stimulation of peripheral blood mononuclear cells (PBMCs) with Hp:Hb complexes was performed and cytokine release in corresponding supernatants were measured using cytometric bead array. The results revealed a higher Hp2-2 phenotype prevalence in the Fulani. The Hp2-2 phenotype was associated with a higher susceptibility to P. falciparum infection in Dogon, but not in Fulani. In concordance with previous studies, Fulani showed increased inflammatory mediators (IL-6, IFN-γ) and additionally also increased sCD163 levels compared to Dogon, irrespective of infection. Furthermore, infected individuals showed elevated sCD163 levels compared to uninfected individuals, in both Fulani and Dogon. Multiple regression analysis revealed that the Hp1-1 phenotype was associated with higher levels of TNF and IFN-γ, as compared to the Hp2-2 phenotype. In vitro stimulation of PBMCs with Hb:Hp1-1 complexes resulted in a pro-inflammatory cytokine profile, whilst stimulation with Hb:Hp2-2 complexes showed a more balanced profile. Ethnicity might be an important confounder on the Hp phenotype-dependent susceptibility to malaria and future studies could consider taking this into account when designing new immunological studies. Although, the relatively small sample size used in this study warrens for precautions in the interpretation of the data and these findings should ideally be validated in a bigger cohort.


Meta-analysis of methods used to assess implant stability.
To evaluate correlations between Periotest, cutting torque or insertion torque, and/or the reverse torque test with resonance frequency analysis measurements. The combined search terms dental implant, primary stability, resonance frequency analysis, removal torque, vibration analysis, torque, Periotest, cutting torque, and insertion torque and specific inclusion and exclusion criteria were used to retrieve eligible articles published between 1998 and 2008 (up to April 1) from PubMed, EMBASE, OVID, The Cochrane Library databases, and 10 journals by hand-searching. The pooled P value was calculated with the Fisher method. The correlation coefficients (r) were combined using fixed and random effect models. Heterogeneity of the studies was tested by performing the homogeneity test. Fifty articles were identified, but 3 studies were excluded because of limitations in study design. Studies that fulfilled the inclusion criteria fell into a variety of categories (11 human cadaver, 16 clinical, 15 animal, and 5 in vitro studies). Assessment of P values revealed that correlation between cutting torque or insertion torque and resonance frequency analysis was significant (6 studies; P = .0022). Correlation between Periotest and cutting torque or insertion torque was significant (1 study; P = .015), and correlations between resonance frequency analysis and reverse torque test values (1 study; P = .319; r = 0.405) and Periotest and resonance frequency analysis (P = .28) were insignificant. Nine articles provided r values, and cutting torque or insertion torque and resonance frequency analysis showed a statistically significant and direct relationship of 55.4% (P = .000). One study provided an r value of -0.149. A statistically significant and direct relationship of 87.6% was found between cutting torque or insertion torque and reverse torque (2 studies; P = .000). This meta-analysis of 47 studies showed a statistically significant correlation between cutting torque or insertion torque and resonance frequency analysis.


Drug-Eluting Medical Textiles: From Fiber Production and Textile Fabrication to Drug Loading and Delivery.
Drug-eluting medical textiles have recently gained great attention to be used in different applications due to their cost effectiveness and unique physical and chemical properties. Using various fiber production and textile fabrication technologies, fibrous constructs with the required properties for the target drug delivery systems can be designed and fabricated. This review summarizes the current advances in the fabrication of drug-eluting medical textiles. Different fiber production methods such as melt-, wet-, and electro-spinning, and textile fabrication techniques such as knitting and weaving are explained. Moreover, various loading processes of bioactive agents to obtain drug-loaded fibrous structures with required physicochemical and morphological properties, drug delivery mechanisms, and drug release kinetics are discussed. Finally, the current applications of drug-eluting fibrous systems in wound care, tissue engineering, and transdermal drug delivery are highlighted.


Enhancing Precision in L-band Electron Paramagnetic Resonance Tooth Dosimetry: Incorporating Digital Image Processing and Radiation Therapy Plans for Geometric Correction.
Following unforeseen exposure to radiation, quick dose determination is essential to prioritize potential patients that require immediate medical care. L-band electron paramagnetic resonance tooth dosimetry can be efficiently used for rapid triage as this poses no harm to the human incisor, although geometric variations among human teeth may hinder accurate dose estimation. Consequently, we propose a practical geometric correction method using a mobile phone camera. Donated human incisors were irradiated with calibrated 6-MV photon beam irradiation, and dose-response curves were developed by irradiation with a predetermined dose using custom-made poly(methyl methacrylate) slab phantoms. Three radiation treatment plans for incisors were selected and altered to suit the head phantom. The mean doses on tooth structures were calculated using a commercial treatment planning system, and the electron paramagnetic resonance signals of the incisors were measured. The enamel area was computed from camera-acquired tooth images. The relative standard uncertainty was rigorously estimated both with and without geometric correction. The effects on the electron paramagnetic resonance signal caused by axial and rotational movements of tooth samples were evaluated through finite element analysis. The mean absolute deviations of mean doses both with and without geometric correction showed marginal improvement. The average relative differences without and with geometric correction significantly decreased from 21.0% to 16.8% (p = 0.01). The geometric correction method shows potential in improving dose precision measurement with minimal delay. Furthermore, our findings demonstrated the viability of using treatment planning system doses in dose estimation for L-band electron paramagnetic resonance tooth dosimetry.


Time and metamorphic petrology: calcite to aragonite experiments.
Although the equilibrium phase relations of many mineral systems are generally well established, the rates of transformations, particularly in polycrystalline rocks, are not. The results of experiments on the calcite to aragonite transformation in polycrystalline marble are different from those for earlier experiments on powdered and single-crystal calcite. The transformation in the polycrystalline samples occurs by different mechanisms, with a different temperature dependence, and at a markedly slower rate. This work demonstrates the importance of kinetic studies on fully dense polycrystalline aggregates for understanding mineralogic phase changes in nature. Extrapolation of these results to geological time scales suggests that transformation of calcite to aragonite does not occur in the absence of volatiles at temperatures below 200 degrees C. Kinetic hindrance is likely to extend to higher temperatures in more complex transformations.


Understanding of and attitudes to genetic testing for inherited retinal disease: a patient perspective.
The views of people with inherited retinal disease are important to help develop health policy and plan services. This study aimed to record levels of understanding of and attitudes to genetic testing for inherited retinal disease, and views on the availability of testing. Telephone questionnaires comprising quantitative and qualitative items were completed with adults with inherited retinal disease. Participants were recruited via postal invitation (response rate 48%), approach at clinic or newsletters of relevant charitable organisations. Questionnaires were completed with 200 participants. Responses indicated that participants' perceived understanding of genetic testing for inherited retinal disease was variable. The majority (90%) considered testing to be good/very good and would be likely to undergo genetic testing (90%) if offered. Most supported the provision of diagnostic (97%) and predictive (92%) testing, but support was less strong for testing as part of reproductive planning. Most (87%) agreed with the statement that testing should be offered only after the individual has received genetic counselling from a professional. Subgroup analyses revealed differences associated with participant age, gender, education level and ethnicity (p<0.02). Participants reported a range of perceived benefits (eg, family planning, access to treatment) and risks (eg, impact upon family relationships, emotional consequences). Adults with inherited retinal disease strongly support the provision of publicly funded genetic testing. Support was stronger for diagnostic and predictive testing than for testing as part of reproductive planning.


Delivery should happen soon and my pain will be reduced: understanding women's perception of good delivery care in India.
Understanding a woman's perspective and her needs during childbirth and addressing them as part of quality-improvement programmes can make delivery care safe, affordable, and respectful. It has been pointed out that the patient's judgement on the quality and goodness of care is indispensible to improving the management of healthcare systems. The objective of the study is to understand the aspects of care that women consider important during childbirth. Individual in-depth interviews (IDIs) and focus-group discussions (FGDs) with women who recently delivered were the techniques used. Seventeen IDIs and four FGDs were conducted in Jharkhand state in east India between January and March 2012. Women who had normal deliveries with live births at home and in primary health centres were included. To minimise recall bias, interviews were conducted within 42 days of childbirth. Using the transcripts of interviews, the data were analysed thematically. Aspects of care most commonly cited by women to be important were: availability of health providers and appropriate medical care (primarily drugs) in case of complications; emotional support; privacy; clean place after delivery; availability of transport to reach the institution; monetary incentives that exceed expenses; and prompt care. Other factors included kind interpersonal behaviour, cognitive support, faith in the provider's competence, and overall cleanliness of the facility and delivery room. Respondents belonging to low socio-economic strata with basic literacy levels might not understand appropriate clinical aspects of care, but they want care that is affordable and accessible, along with privacy and emotional support during delivery. The study highlighted that healthcare quality-improvement programmes in India need to include non-clinical aspects of care as women want to be treated humanely during delivery--they desire respectful treatment, privacy, and emotional support. Further research into maternal satisfaction could be made more policy relevant by assessing the relative strength of various factors in influencing maternal satisfaction; this could help in prioritising appropriate interventions for improved quality of care (QoC).


Quasistationarity in a model of long-range interacting particles moving on a sphere.
We consider a long-range interacting system of N particles moving on a spherical surface under an attractive Heisenberg-like interaction of infinite range and evolving under deterministic Hamilton dynamics. The system may also be viewed as one of globally coupled Heisenberg spins. In equilibrium, the system has a continuous phase transition from a low-energy magnetized phase, in which the particles are clustered on the spherical surface, to a high-energy homogeneous phase. The dynamical behavior of the model is studied analytically by analyzing the Vlasov equation for the evolution of the single-particle distribution and numerically by direct simulations. The model is found to exhibit long-lived nonmagnetized quasistationary states (QSSs) which in the thermodynamic limit are dynamically stable within an energy range where the equilibrium state is magnetized. For finite N, these states relax to equilibrium over a time that increases algebraically with N. In the dynamically unstable regime, nonmagnetized states relax fast to equilibrium over a time that scales as lnN. These features are retained in presence of a global anisotropy in the magnetization.


Workplace tobacco interventions.
Health promotion programs are becoming an integral part of work site activities. Recent data from Indiana businesses suggest that smoking is a leading concern. An objective has been adopted by the Indiana Chamber of Commerce that states that the number of employers with work site smoking cessation policies should increase. Smoking control and cessation programs implemented in industry have contributed to a decrease in the number of smokers and in the health risks of nonsmokers exposed to environmental tobacco smoke. This report describes the effectiveness of work site smoking control programs.


Long-term results after surgical treatment of the dominant lung adenocarcinoma associated with ground-glass opacities.
To evaluate the effect of first-time and eventual reiterative surgery on overall survival (OS) and disease-free survival (DFS) in Caucasian patients affected by an invasive adenocarcinoma (ADC) with at least another ground-glass opacity (GGO). We analysed 47 patients operated on for lung ADC, identified as main cancer (MC), with at least one synchronous GGO, from January 2003 to March 2017. Characteristics associated with the evolution of GGOs were investigated with logistic regression and overall and DFS were evaluated with Kaplan-Meier method. Forty-two (89%) patients received an anatomic resection of the MC, 5 patients were treated by a single or multiple wedge resections. In total, 9 (19.1%) patients had all the lesions resected undergoing simultaneous resection of ipsilateral GGOs at first surgery while the remaining 38 (80.9%) patients still had at least one GGO that was followed up by serial CT scan. At the median follow-up of 41 months, GGO evolved in 16 (42.1%) patients. The presence of solid component at the initial CT scan was the only risk factor for evolution of the GGO. Thirteen patients underwent surgical resection showing an invasive ADC in 9 patients, MIA in 3 and AIS in 1. New GGOs developed in 7 (14.9%) patients, in which three underwent surgery showing the presence of solid ADC, MIA and AAH. OS rate at 5 years was 97.4%. DFS at 3 years was 82% and was significantly influenced by the stage of MC. Patients affected by an invasive ADC with at least another GGO nodule enjoy good OS and DFS with a surgical reiterative approach. Part-solid GGO is associated with GGO progression requiring treatment, but OS is not influenced by the new onset or evolution of GGOs. DFS is affected by the stage of the MC which dictates the treatment strategy.


Simulation and verification of hydraulic properties and organic matter degradation in sand filters for greywater treatment.
To evaluate the treatment performance of vertical flow sand filters, the HYDRUS wetland module was used to simulate treatment in an experimental set-up. The laboratory filters were intermittently dosed with artificial greywater at a hydraulic loading rate of 0.032 m³ m⁻² day⁻¹ and an organic loading rate of 0.014 kg BOD5 m⁻² day⁻¹. The hydraulic properties of the filter were characterised, as were inflow and outflow concentrations of chemical oxygen demand (COD), biochemical oxygen demand (BOD), ammonia, nitrate and total nitrogen. The inverse simulation function of the HYDRUS software was used to calibrate the water flow model. The observed effect of water flowing faster along the column wall was included in the inverse simulations. The biokinetic model was calibrated by fitting heterotrophic biomass growth to measurements of potential respiration rate. Emphasis was put on simulating outflow concentrations of organic pollutants. The simulations were conducted using three models of varying degree of calibration effort and output accuracy. The effluent concentration was 245 mg COD L⁻¹ for the laboratory filters, 134 mg COD mg L⁻¹ for the model excluding wall flow effects and 338 mg COD mg L⁻¹ for the model including wall flow effects.


Lamotrigine pharmacokinetics in patients receiving felbamate.
Drug interactions can significantly complicate the management of patients receiving multiple medications. It is essential therefore that potential pharmcokinetic interactions be evaluated as new antiepileptic medications are introduced. Lamotrigine (LTG) is a recently marketed medication whose pharmacokinetics are significantly influenced by concomitant drugs. Felbamate (FBM), another relatively new antiepileptic agent has been associated with multiple interactions including both enzyme induction and inhibition. The purpose of the present pilot study was to evaluate potential differences in lamotrigine kinetics in six patients concomitantly receiving FBM compared to five patients receiving lamotrigine as monotherapy. There was no statistically significant differences in either apparent LTG oral clearance (0.026 +/- 0.005 vs. 0.024 +/- 0.01 l/kg per h, respectively), or in mean elimination half-life (33.7 +/- 7.5 vs. 40.2 +/- 15.05 h, respectively). Oral clearance values in our patients are also consistent with data reported previously in the literature. Data from this pilot study suggest that a marked effect of FBM upon lamotrigine pharmacokinetics is unlikely.


Studying NB-LRR immune receptor localization by agroinfiltration transient expression.
NB-LRR immune receptors in plants play dual roles as sentries and as activators of defense. The site in the cell where these activities take place can be different for different NB-LRRs. Furthermore, recognition and defense activation can occur in distinct subcellular compartments. Therefore, determining the subcellular localization of NB-LRRs is a key step toward understanding how they function. Recent advances in confocal microscopy enable high-resolution imaging of proteins in live cells. Agroinfiltration in the Nicotiana benthamiana model plant system is a convenient way of expressing proteins for localization studies. This chapter explains how to use N. benthamiana to transiently express NB-LRRs for confocal fluorescence microscopy.


Overlaps Between CDS Regions of Protein-Coding Genes in the Human Genome: A Case Study on the NR1D1-THRA Gene Pair.
For several decades, it has been known that a substantial number of genes within human DNA exhibit overlap; however, the biological and evolutionary significance of these overlaps remain poorly understood. This study focused on investigating specific instances of overlap where the overlapping DNA region encompasses the coding DNA sequences (CDSs) of protein-coding genes. The results revealed that proteins encoded by overlapping CDSs exhibit greater disorder than those from nonoverlapping CDSs. Additionally, these DNA regions were identified as GC-rich. This could be partially attributed to the absence of stop codons from two distinct reading frames rather than one. Furthermore, these regions were found to harbour fewer single-nucleotide polymorphism (SNP) sites, possibly due to constraints arising from the overlapping state where mutations could affect two genes simultaneously.While elucidating these properties, the NR1D1-THRA gene pair emerged as an exceptional case with highly structured proteins and a distinctly conserved sequence across eutherian mammals. Both NR1D1 and THRA are nuclear receptors lacking a ligand-binding domain at their C-terminus, which is the region where these gene pairs overlap. The NR1D1 gene is involved in the regulation of circadian rhythm, while the THRA gene encodes a thyroid hormone receptor, and both play crucial roles in various physiological processes. This study suggests that, in addition to their well-established functions, the specifically overlapping CDS regions of these genes may encode protein segments with additional, yet undiscovered, biological roles.


Transvenous to arterial Onyx embolization.
Transarterial infusion of Onyx is now widely used in the treatment of arteriovenous malformations (AVMs) and dural arteriovenous fistulas (DAVFs). Transvenous Onyx embolization is rarely performed, and usually in cases of carotid cavernous fistulas. We describe a novel technique of transvenous Onyx embolization in the treatment of three high risk DAVFs and one vein of Galen (VOG) AVM. Cases were evaluated with specific attention to technical caveats, clinical indications, and complications. Patients were treated during 2011-2012 by two operators (FCA and CGM). Standard transfemoral venous approaches were employed in order to obtain access into the draining venous system in all cases. Four female patients, aged 5 months to 68 years, were treated. Two patients, one presenting with headache and the other with a bruit, demonstrated high risk DAVFs involving the transverse-sigmoid sinuses. One patient, who was asymptomatic, manifested a high risk ethmoidal DAVF that was discovered incidentally during treatment of a basilar aneurysm. The final patient presented with symptoms of a pineal region mass and was found to have a VOG AVM. In all cases, a microcatheter was navigated through the draining venous pouch and into the ostium of an arterial feeder. A single Onyx infusion from this position filled multiple arterial feeders and resulted in cures in the first three patients and a significant reduction in AVM nidus in the fourth patient. No complications occurred. Transvenous Onyx infusion into a single arterial feeder can produce retrograde embolization of numerous other arterial feeders supplying high risk DAVFs and complex cerebral AVMs. This technique appears to be safe and potentially curative in select cases.


Gender discrimination in the United States: Experiences of women.
To examine reported experiences of gender discrimination and harassment among US women. Data come from a nationally representative, probability-based telephone survey of 1596 women, conducted January-April 2017. We calculated the percentages of women reporting gender discrimination and harassment in several domains, including health care. We used logistic regression to examine variation in experiences among women by race/ethnicity and sexual orientation/gender identity. Sizable fractions of women experience discrimination and harassment, including discrimination in health care (18 percent), equal pay/promotions (41 percent), and higher education (20 percent). In adjusted models, Native American, black, and Latina women had higher odds than white women of reporting gender discrimination in several domains, including health care. Latinas' odds of health care avoidance versus whites was (OR [95% CI]) 3.69 (1.59, 8.58), while blacks' odds of discrimination in health care visits versus whites was 2.00 [1.06, 3.74]. Lesbian, gay, bisexual, transgender, and queer (LGBTQ) women had higher odds of reporting sexual harassment (2.16 [1.06, 4.40]) and violence (2.71 [1.43, 5.16]) against themselves or female family members than non-LGBTQ women. Results suggest that discrimination and harassment are widely experienced by women across multiple domains of their lives, particularly those who are a racial/ethnic minority or LGBTQ. Further policy and programmatic efforts beyond current legal protections for women are needed to meaningfully reduce these negative experiences, as they impact women's health care and their lives overall.


Associations Between Core Temperature Disorders and Outcomes of Pediatric Intensive Care Unit Patients.
The few studies of associations between fever and outcomes in pediatric intensive care unit (PICU) patients have conflicting findings. Associations between hypothermia and patient outcomes have not been studied. To investigate the incidence and characteristics of fever and hypothermia and their associations with adverse outcomes among PICU patients. Patients consecutively admitted to 2 PICUs in a 2-year period were prospectively studied. Core temperature was mainly measured by rectal or axillary thermometry. Fever and hypothermia were defined as core temperatures of greater than 38.0 °C and less than 36.0 °C, respectively. Prolonged mechanical ventilation, prolonged PICU stay, and PICU mortality were the adverse patient outcomes studied. Associations between patient outcomes and core temperature disorders were evaluated with univariate comparisons and multivariate analyses. Of 545 patients enrolled, fever occurred in 299 (54.9%) and hypothermia occurred in 161 (29.5%). Both temperature disorders were independently associated with prolonged mechanical ventilation and prolonged PICU stay (P < .001) but not with PICU mortality. Late onset of fever (P < .001) and hypothermia (P = .009) were independently associated with prolonged mechanical ventilation, fever magnitude and duration (both P < .001) were independently associated with prolonged PICU stay, and fever magnitude (P < .001) and infectious cause of hypothermia (P= .01) were independently associated with higher PICU mortality. These findings provide evidence that the manifestation and characteristics of fever and hypothermia are independent predictors of adverse outcomes in PICU patients.


Interaction of the ectomesenchyme and optic vesicle in mice.
We investigated eye development in embryos at the 17, 24, and 31 somite-pair stages homozygous for the two genes white (Mi-wh) and fidget (fi) (Mi-wh/Mi-whfi/fi), single homozygotes (fi/fi and Mi-wh/Mi-wh), and normal mice (the inbred C57 BL/Mib strain). The fi gene retards optic-vesicle growth, while the Mi-wh gene inhibits ectomesenchyme migration. It was shown that the mitotic index of the optic-vesicle wall and the retinal rudiment was considerably higher in Mi-wh/Mi-whfi/fi embryos than in fi/fi embryos and lower than in Mi-wh/Mi-wh and +/+ embryos. Ectomesenchyme affected by the white gene stimulated optic-vesicle growth, to some extent suppressing the effect of the fidget gene. The stimulation of optic-vesicle growth in the double homozygotes led to intensified induction of the lens placode. This indicates that the ectomesenchyme affects the optic rudiment at the optic-vesicle stage.


Genetic algorithm-based classifiers fusion for multisensor activity recognition of elderly people.
Activity recognition of an elderly person can be used to provide information and intelligent services to health care professionals, carers, elderly people, and their families so that the elderly people can remain at homes independently. This study investigates the use and contribution of wrist-worn multisensors for activity recognition. We found that accelerometers are the most important sensors and heart rate data can be used to boost classification of activities with diverse heart rates. We propose a genetic algorithm-based fusion weight selection (GAFW) approach which utilizes GA to find fusion weights. For all possible classifier combinations and fusion methods, the study shows that 98% of times GAFW can achieve equal or higher accuracy than the best classifier within the group.


Pharmacological Extracts and Molecules from Virola Species: Traditional Uses, Phytochemistry, and Biological Activity.
Virola is the largest genus of Myristicaceae in America, comprising about 60 species of medium-sized trees geographically spread from Mexico to southern Brazil. The plant species of this genus have been widely used in folk medicine for the treatment of several ailments, such as rheumatic pain, bronchial asthma, tumors in the joints, intestinal worms, halitosis, ulcers, and multiple infections, due to their pharmacological activity. This review presents an updated and comprehensive summary of Virola species, particularly their ethnomedicinal uses, phytochemistry, and biological activity, to support the safe medicinal use of plant extracts and provide guidance for future research. The Virola spp.'s ethnopharmacology, including in the treatment of stomach pain and gastric ulcers, as well as antimicrobial and tryponosomicidal activities, is attributable to the presence of a myriad of phytoconstituents, such as flavonoids, tannins, phenolic acids, lignans, arylalkanones, and sitosterol. Hence, such species yield potential leads or molecular scaffolds for the development of new pharmaceutical formulations, encouraging the elucidation of not-yet-understood action mechanisms and ascertaining their safety for humans.


It's Not "Just A Finger".
To provide a source of information to the athletic trainer on injuries to the fingers; to detail the pertinent anatomy of the finger and explain the techniques of a proper examination; and to discuss treatment and possible outcomes. Injuries to the fingers are often dismissed as inconsequential and are usually only touched upon briefly in articles on hand injuries. The article is divided into detailed sections pertaining to anatomy, examination, and specific injuries and their treatment. CLINICAL ADVANTAGE: This article provides a concise source of information to athletic trainers on finger injuries. They need not refer to multiple volumes of journals and texts to find the specific information they seek.


Galactose differentially modulates lunatic and manic fringe effects on Delta1-induced NOTCH signaling.
NOTCH signaling induced by Delta1 (DLL1) and Jagged1 (JAG1) NOTCH ligands is modulated by the β3N-acetylglucosaminyl transferase Fringe. LFNG (Lunatic Fringe) and MFNG (Manic Fringe) transfer N-acetylglucosamine (GlcNAc) to O-fucose attached to EGF-like repeats of NOTCH receptors. In co-culture NOTCH signaling assays, LFNG generally enhances DLL1-induced, but inhibits JAG1-induced, NOTCH signaling. In mutant Chinese hamster ovary (CHO) cells that do not add galactose (Gal) to the GlcNAc transferred by Fringe, JAG1-induced NOTCH signaling is not inhibited by LFNG or MFNG. In mouse embryos lacking B4galt1, NOTCH signaling is subtly reduced during somitogenesis. Here we show that DLL1-induced NOTCH signaling in CHO cells was enhanced by LFNG, but this did not occur in either Lec8 or Lec20 CHO mutants lacking Gal on O-fucose glycans. Lec20 mutants corrected with a B4galt1 cDNA became responsive to LFNG. By contrast, MFNG promoted DLL1-induced NOTCH signaling better in the absence of Gal than in its presence. This effect was reversed in Lec8 cells corrected by expression of a UDP-Gal transporter cDNA. The MFNG effect was abolished by a DDD to DDA mutation that inactivates MFNG GlcNAc transferase activity. The binding of soluble NOTCH ligands and NOTCH1/EGF1-36 generally reflected changes in NOTCH signaling caused by LFNG and MFNG. Therefore, the presence of Gal on O-fucose glycans differentially affects DLL1-induced NOTCH signaling modulated by LFNG versus MFNG. Gal enhances the effect of LFNG but inhibits the effect of MFNG on DLL1-induced NOTCH signaling, with functional consequences for regulating the strength of NOTCH signaling.


Prediction of hepatotoxicity for drugs using human pluripotent stem cell-derived hepatocytes.
Drug-induced liver toxicity is a main reason for withdrawals of new drugs in late clinical phases and post-launch of the drugs. Thus, hepatotoxicity screening of drug candidates in pre-clinical stage is important for reducing drug attrition rates during the clinical development process. Here, we show commercially available hepatocytes that could be used for early toxicity evaluation of drug candidates. From our hepatic differentiation technology, we obtained highly pure (≥98%) hepatocytes from human embryonic stem cells (hESCs) having mature phenotypes and similar gene expression profiles with those of primary human tissues. Furthermore, we optimized 96-well culture condition of hESC-derived hepatocytes suitable for toxicity tests in vitro. To this end, we demonstrated the efficacy of our optimized hepatocyte model for predicting hepatotoxicity against the Chinese herbal medicines and showed that toxicity patterns from our hepatocyte model was similar to those of human primary cultured hepatocytes. We conclude that toxicity test using our hepatocyte model could be a good alternative cell source for pre-clinical study to predict potential hepatotoxicity in drug discovery industries.


What do emergency medicine learners want from their teachers? A multicenter focus group analysis.
To the best of the authors' knowledge, there are no reports describing what learners believe are good emergency medicine (EM) teaching practices. EM faculty developers are compromised by this lack of knowledge about what EM learners appreciate in their teachers. To determine what Canadian EM learners consider to be good prerequisites and strategies for effective teaching in the emergency department (ED). Clinical clerks and residents from the Canadian College of Family Physicians, Emergency Medicine certification [CCFP(EM)] fellowship program, the Royal College of Physicians and Surgeons of Canada, Emergency Medicine certification [FRCP(EM)] fellowship program, and off-service programs from all five Ontario medical schools participated in monitored focus-group sessions. Conversations were recorded, transcribed by a third party, and coded by two independent assessors using standard grounded theory methods. The text was categorized based on the final code into basic themes and specific qualifiers, which were then sorted by frequency of mention in the focus groups. Results are presented in descriptive fashion. Twenty-eight learners participated. They identified 14 major principles for good EM teaching, and a further 30 specific qualifiers. The top five principles were: "has a positive teacher attitude," "takes time to teach," "uses teachable moments well," "tailors teaching to the learner," and "gives appropriate feedback." Agreement on classification of ideas was 86%. Learners are sensitive to the constraints of the ED teaching environment, and have consistent views about good ED teaching practices. Among 14 general principles identified, "takes time to teach," "gives feedback," "tailors teaching to the learner," "uses teachable moments," and "has a good teacher attitude" were the most commonly reported.


Equally good fixation of cemented and uncemented cups in total trapeziometacarpal joint prostheses. A randomized clinical RSA study with 2-year follow-up.
Cup failure is a recognized problem in total trapeziometacarpal (TM) joint prostheses; it may be related to poor fixation, which can be revealed by radiostereometric analysis (RSA). We compared the early implant migration of cemented trapezium cups to that of uncemented screw cups. In a prospective, parallel-group, randomized patient-blinded clinical trial, we included 32 hands in 28 patients (5 males) with a mean age of 58 (40-77) years and with Eaton stage-2 or -3 osteoarthritis of the trapeziometacarpal joint. Patients were randomized to surgery with a cemented DLC all-polyethylene cup (C) (n = 16) or an uncemented hydroxyapatite-coated chrome-cobalt Elektra screw cup (UC) (n = 16). Uncemented cups were inserted without threading of the bone. Stereoradiographs for evaluation of cup migration (primary effect size) and DASH and pain scores were obtained during 2 years of follow-up. The 2-year total translation (TT) was similar (p = 0.2): 0.24 mm (SD 0.10) for the C (n = 11) and 0.19 mm (SD 0.16) for the UC (n = 11). Variances were similar (p = 0.4). Judged by RSA, 2 UC cups and 1 C cup became loose (TT > 1 mm). Both UC cups were found to be loose at revision. Grip strength, pain, and DASH scores were similar between groups at all measurement points. Early implant fixation and clinical outcome were equally good with both cup designs. This is the first clinical RSA study on trapezium cups, and the method appears to be clinically useful for detection of loose implants.


Accelerated Monte Carlo Rendering of Finite-Time Lyapunov Exponents.
Time-dependent fluid flows often contain numerous hyperbolic Lagrangian coherent structures, which act as transport barriers that guide the advection. The finite-time Lyapunov exponent is a commonly-used approximation to locate these repelling or attracting structures. Especially on large numerical simulations, the FTLE ridges can become arbitrarily sharp and very complex. Thus, the discrete sampling onto a grid for a subsequent direct volume rendering is likely to miss sharp ridges in the visualization. For this reason, an unbiased Monte Carlo-based rendering approach was recently proposed that treats the FTLE field as participating medium with single scattering. This method constructs a ground truth rendering without discretization, but it is prohibitively slow with render times in the order of days or weeks for a single image. In this paper, we accelerate the rendering process significantly, which allows us to compute video sequence of high-resolution FTLE animations in a much more reasonable time frame. For this, we follow two orthogonal approaches to improve on the rendering process: the volumetric light path integration in gradient domain and an acceleration of the transmittance estimation. We analyze the convergence and performance of the proposed method and demonstrate the approach by rendering complex FTLE fields in several 3D vector fields.


Hedgehog signaling regulates telomerase reverse transcriptase in human cancer cells.
The Hedgehog (HH) signaling pathway is critical for normal embryonic development, tissue patterning and cell differentiation. Aberrant HH signaling is involved in multiple human cancers. HH signaling involves a multi-protein cascade activating the GLI proteins that transcriptionally regulate HH target genes. We have previously reported that HH signaling is essential for human colon cancer cell survival and inhibition of this signal induces DNA damage and extensive cell death. Here we report that the HH/GLI axis regulates human telomerase reverse transcriptase (hTERT), which determines the replication potential of cancer cells. Suppression of GLI1/GLI2 functions by a C-terminus truncated GLI3 repressor mutant (GLI3R), or by GANT61, a pharmacological inhibitor of GLI1/GLI2, reduced hTERT protein expression in human colon cancer, prostate cancer and Glioblastoma multiforme (GBM) cell lines. Expression of an N-terminus deleted constitutively active mutant of GLI2 (GLI2ΔN) increased hTERT mRNA and protein expression and hTERT promoter driven luciferase activity in human colon cancer cells while GANT61 inhibited hTERT mRNA expression and hTERT promoter driven luciferase activity. Chromatin immunoprecipitation with GLI1 or GLI2 antibodies precipitated fragments of the hTERT promoter in human colon cancer cells, which was reduced upon exposure to GANT61. In contrast, expression of GLI1 or GLI2ΔN in non-malignant 293T cells failed to alter the levels of hTERT mRNA and protein, or hTERT promoter driven luciferase activity. Further, expression of GLI2ΔN increased the telomerase enzyme activity, which was reduced by GANT61 administration in human colon cancer, prostate cancer, and GBM cells. These results identify hTERT as a direct target of the HH signaling pathway, and reveal a previously unknown role of the HH/GLI axis in regulating the replication potential of cancer cells. These findings are of significance in understanding the important regulatory mechanisms that determine the functions of HH/GLI signaling in cancer cells.


Enhanced biodegradation of atrazine by bacteria encapsulated in organically modified silica gels.
Biodegradation by cells encapsulated in silica gel is an economical and environmentally friendly method for the removal of toxic chemicals from the environment. In this work, recombinant E. coli expressing atrazine chlorohydrolase (AtzA) were encapsulated in organically modified silica (ORMOSIL) gels composed of TEOS, silica nanoparticles (SNPs), and either phenyltriethoxysilane (PTES) or methyltriethoxysilane (MTES). ORMOSIL gels adsorbed much higher amounts of atrazine than the hydrophilic TEOS gels. The highest amount of atrazine adsorbed by ORMOSIL gels was 48.91×10-3μmol/mlgel, compared to 8.71×10-3μmol/mlgel by the hydrophilic TEOS gels. Atrazine biodegradation rates were also higher in ORMOSIL gels than the TEOS gels, mainly due to co-localization of the hydrophobic substrate at high concentrations in close proximity of the encapsulated bacteria. A direct correlation between atrazine adsorption and biodegradation was observed unless biodegradation decreased due to severe phase separation. The optimized PTES and MTES gels had atrazine biodegradation rates of 0.041±0.003 and 0.047±0.004μmol/mlgel, respectively. These rates were approximately 80% higher than that measured in the TEOS gel. This study showed for the first time that optimized hydrophobic gel material design can be used to enhance both removal and biodegradation of hydrophobic chemicals.


Involvement of CX3CL1 in the Migration of Osteoclast Precursors Across Osteoblast Layer Stimulated by Interleukin-1ß.
The trigger for bone remodeling is bone resorption by osteoclasts. Osteoclast differentiation only occurs on the old bone, which needs to be repaired under physiological conditions. However, uncontrolled bone resorption is often observed in pro-inflammatory bone diseases, such as rheumatoid arthritis. Mature osteoclasts are multinuclear cells that differentiate from monocyte/macrophage lineage cells by cell fusion. Although Osteoclast precursors should migrate across osteoblast layer to reach bone matrix before maturation, the underlying mechanisms have not yet been elucidated in detail. We herein found that osteoclast precursors utilize two routes to migrate across osteoblast layer by confocal- and electro-microscopic observations. The osteoclast supporting activity of osteoblasts inversely correlated with osteoblast density and was positively related to the number of osteoclast precursors under the osteoblast layer. Osteoclast differentiation was induced by IL-1ß, but not by PGE2 in high-density osteoblasts. Osteoblasts and osteoclast precursors expressed CX3CL1 and CX3CR1, respectively, and the expression of CX3CL1 increased in response to interleukin-1ß. An anti-CX3CL1-neutralizing antibody inhibited the migration of osteoclast precursors and osteoclast differentiation. These results strongly suggest the involvement of CX3CL1 in the migration of osteoclast precursors and osteoclastogenesis, and will contribute to the development of new therapies for bone diseases. J. Cell. Physiol. 232: 1739-1745, 2017. © 2016 Wiley Periodicals, Inc.


Herpes simplex virus US3 tegument protein inhibits Toll-like receptor 2 signaling at or before TRAF6 ubiquitination.
Herpes simplex virus (HSV) has evolved multiple strategies to modulate host immune responses. In a screen of HSV open reading frames to identify additional HSV-encoded proteins that affect NF-κB signaling, we identified the viral US3 tegument protein as an inhibitor of NF-κB signaling. We found that the US3 protein is required for inhibition of TLR2 signaling induced by viral infection and that this inhibition occurs at very early times post-infection. Expression of US3 in transfected cells inhibits TLR2 signaling induced by Zymosan, and this inhibition occurs at or downstream of MyD88 and upstream of p65. Polyubiquitination of TRAF6 is critical for its function in TLR2 signaling. Using US3-null and US3 kinase-defective mutant viruses, we demonstrate that HSV US3 reduces TRAF6 polyubiquitination and that the kinase activity of US3 is necessary for this effect. Therefore, US3 is necessary and sufficient for inhibiting TLR2 signaling at or before the stage of TRAF6 ubiquitination.


Predictors of mortality and loss to follow-up among drug resistant tuberculosis patients in Oromia Hospitals, Ethiopia: A retrospective follow-up study.
Drug resistance tuberculosis (DR-TB) patients' mortality and loss to follow-up (LTF) from treatment and care is a growing worry in Ethiopia. However, little is known about predictors of mortality and LTF among drug-resistant tuberculosis patients in Oromia region, Ethiopia. The current study aimed to identify predictors of mortality and loss to follow-up among drug resistance tuberculosis patients in Oromia Hospitals, Ethiopia. A retrospective follow up study was carried out from 01 November 2012 to 31 December 2017 among DR-TB patients after calculating sample size using single proportion population formula. Mean, median, Frequency tables and bar charts were used to describe patients' characteristics in the cohort. The Kaplan-Meier curve was used to estimate the probability of death and LTF after the treatment was initiated. The log-rank test was used to compare time to death and time to LTF. The Cox proportional hazard model was used to determine predictors of mortality and LTF after DR-TB diagnosis. The Crude and adjusted Cox proportional hazard ratio was used to measure the strength of association whereas p-value less than 0.05 were used to declare statistically significant predictors. A total of 406 DR-TB patients were followed for 7084 person-months observations. Among the patients, 71 (17.5%) died and 32 (7.9%) were lost to follow up (LTF). The incidence density of death and LTF in the cohort was 9.8 and 4.5 per 1000 person-months, respectively. The median age of the study participants was 28 years (IQR: 27.1, 29.1). The overall cumulative survival probability of patients at the end of 24 months was 77.5% and 84.5% for the mortality and LTF, respectively. The independent predictors of death was chest radiographic findings (AHR = 0.37, 95% CI: 0.17-0.79) and HIV serostatus 2.98 (95% CI: 1.72-5.19). Drug adverse effect (AHR = 6.1; 95% CI: 2.5, 14.34) and culture test result (AHR = 0.1; 95% CI: 0.1, 0.3) were independent predictors of LTF. This study concluded that drug-resistant tuberculosis mortality and LTF remains high in the study area. Continual support of the integration of TB/HIV service with emphasis and work to identified predictors may help in reducing drug-resistant tuberculosis mortality and LTF.


Analysis of specific serum markers of colon carcinoma using a Bhattacharyya-based support vector machine.
We aimed to evaluate the specificity of 12 tumor markers related to colon carcinoma and identify the most sensitive index. Bhattacharyya distance was used to evaluate the index. Then, different index combinations were used to establish a support vector machine (SVM) diagnosis model of malignant colon carcinoma. The accuracy of the model was checked. High accuracy was assumed to indicate the high specificity of the index. The Bhattacharyya distances of carcinoembryonic antigen, neuron-specific enolase, alpha-feto protein, and CA724 were the largest, and those of CYFRA21-І, CA125, and UGT1A83 were the second largest. The specificity of the combination of the above seven indexes was higher than that of other combinations, and the accuracy of the established SVM identification model was high. Using Bhattacharyya distance detection and establishing an SVM model based on different serum marker combinations can increase diagnostic accuracy, providing a theoretical basis for application of mathematical models in cancer diagnosis.


[Protein kinase CK2 and cancer: further clues are accumulating].
Protein kinase CK2 appears to interact with different signaling pathways and therefore represents the prototype of a multifunctional protein kinase. The enzyme is highly expressed in most cancers and this higher expression has been tentatively correlated with the involvement of CK2 in the promotion of specific phases of the cell cycle. This review summarizes our current knowledge on how CK2 may also contribute to tumorigenesis through its direct interaction with the cell-survival circuitry.


Age-related differences in lean mass, protein synthesis and skeletal muscle markers of proteolysis after bed rest and exercise rehabilitation.
Bed rest-induced muscle loss and impaired muscle recovery may contribute to age-related sarcopenia. It is unknown if there are age-related differences in muscle mass and muscle anabolic and catabolic responses to bed rest. A secondary objective was to determine if rehabilitation could reverse bed rest responses. Nine older and fourteen young adults participated in a 5-day bed rest challenge (BED REST). This was followed by 8 weeks of high intensity resistance exercise (REHAB). Leg lean mass (via dual-energy X-ray absorptiometry; DXA) and strength were determined. Muscle biopsies were collected during a constant stable isotope infusion in the postabsorptive state and after essential amino acid (EAA) ingestion on three occasions: before (PRE), after bed rest and after rehabilitation. Samples were assessed for protein synthesis, mTORC1 signalling, REDD1/2 expression and molecular markers related to muscle proteolysis (MURF1, MAFBX, AMPKα, LC3II/I, Beclin1). We found that leg lean mass and strength decreased in older but not younger adults after bedrest (P < 0.05) and was restored after rehabilitation. EAA-induced mTORC1 signalling and protein synthesis increased before bed rest in both age groups (P < 0.05). Although both groups had blunted mTORC1 signalling, increased REDD2 and MURF1 mRNA after bedrest, only older adults had reduced EAA-induced protein synthesis rates and increased MAFBX mRNA, p-AMPKα and the LC3II/I ratio (P < 0.05). We conclude that older adults are more susceptible than young persons to muscle loss after short-term bed rest. This may be partially explained by a combined suppression of protein synthesis and a marginal increase in proteolytic markers. Finally, rehabilitation restored bed rest-induced deficits in lean mass and strength in older adults.


Functional genomics of calcium channels in human melanoma cells.
Ca(2+)-signaling of human melanoma is in the focus of intensive research since the identification of the role of WNT-signaling in melanomagenesis. Genomic and functional studies pointed to the important role of various Ca(2+) channels in melanoma, but these data were contradictory. In the present study we clearly demonstrate, in a number of different ways including microarray analysis, DNA sequencing and immunocytochemistry, that various human melanoma cell lines and melanoma tissues overexpress ryanodine receptor type 2 (RyR2) and express P2X(7) channel proteins as compared to melanocytes. These channels, although retain some of their usual characteristics and pharmacological properties, display unique features in melanoma cells, including a functional interaction between the two molecules. Unlike P2X(7), RyR2 does not function as a calcium channel. On the other hand, the P2X(7) receptor has an antiapoptotic function in melanoma cells, since ATP-activation suppresses induced apoptosis, while knock down of the gene expression significantly enhances that.


Multi-Imager Compatible, MR Safe, Remote Center of Motion Needle-Guide Robot.
We report the development of a new robotic system for direct image-guided interventions (DIGI; images acquired at the time of the intervention). The manipulator uses our previously reported pneumatic step motors and is entirely made of electrically nonconductive, nonmetallic, and nonmagnetic materials. It orients a needle-guide with two degrees of freedom (DoF) about a fulcrum point located below the guide using an innovative remote center of motion parallelogram type mechanism. The depth of manual needle insertion is preset with a third DoF, located remotely of the manipulator. Special consideration was given to the kinematic accuracy and the structural stiffness. The manipulator includes registration markers for image-to-robot registration. Based on the images, it may guide needles, drills, or other slender instruments to a target (OD < 10 mm). Comprehensive preclinical tests were performed. The manipulator is MR safe (ASTM F2503-13). Electromagnetic compatibility (EMC) testing (IEC 60601-1-2) of the system shows that it does not conduct or radiate EM emissions. The change in the signal to noise ratio of the MRI due to the presence and motion of the robot in the scanner is below 1%. The structural stiffness at the needle-guide is 33 N/mm. The angular accuracy and precision of the manipulator itself are 0.177° and 0.077°. MRI-guided targeting accuracy and precision in vitro were 1.71 mm and 0.51 mm, at an average target depth of ∼38 mm, with no adjustments. The system may be suitable for DIGI where [mm] accuracy lateral to the needle (2D) or [mm] in 3D is acceptable. The system is also multi-imager compatible and could be used with other imaging modalities.


The Wild West of executive coaching.
Annual spending on executive coaching in the United States is estimated at 1 billion dollars. Yet information about coaching's effectiveness is scarce and unreliable. No one has yet demonstrated conclusively what qualifies an executive coach or what makes one approach to executive coaching better than another. Barriers to entry are nonexistent--many executive coaches know little about business, and some know little about coaching. The coaching certifications offered by various self-appointed bodies are difficult to assess, and methods of measuring return on investment are questionable. But strategic coaching can provide critical help both to individuals and to organizations. In this article, Stratford Sherman, a senior vice president of Executive Coaching Network, and Alyssa Freas, the founder and CEO, explore the popularity of executive coaching and investigate ways to make the most of the experience. They argue that coaching is inevitably a triangular relationship between the client, the "coachee," and the coach. Its purpose is to produce behavioral change and growth in the coachee for the economic benefit of the client. The best way to maximize the likelihood of good results is to qualify all the people involved. Even so, many triangular relationships continue to generate conflict among all three parties. At the most basic level, coaches serve as suppliers of candor, providing leaders with the objective feedback they need to nourish their growth. Coaching gets executives to slow down, gain awareness, and notice the effects of their words and actions. On a larger scale, the best coaching fosters cultural change for the benefit of the entire organization. It provides a disciplined way for businesses to deepen relationships with their most valued employees while also increasing their effectiveness.


Control of neurotransmitter release by two distinct membrane-binding faces of the Munc13-1 C1C2B region.
Munc13-1 plays a central role in neurotransmitter release through its conserved C-terminal region, which includes a diacyglycerol (DAG)-binding C1 domain, a Ca2+/PIP2-binding C2B domain, a MUN domain and a C2C domain. Munc13-1 was proposed to bridge synaptic vesicles to the plasma membrane through distinct interactions of the C1C2B region with the plasma membrane: (i) one involving a polybasic face that is expected to yield a perpendicular orientation of Munc13-1 and hinder release; and (ii) another involving the DAG-Ca2+-PIP2-binding face that is predicted to result in a slanted orientation and facilitate release. Here, we have tested this model and investigated the role of the C1C2B region in neurotransmitter release. We find that K603E or R769E point mutations in the polybasic face severely impair Ca2+-independent liposome bridging and fusion in in vitro reconstitution assays, and synaptic vesicle priming in primary murine hippocampal cultures. A K720E mutation in the polybasic face and a K706E mutation in the C2B domain Ca2+-binding loops have milder effects in reconstitution assays and do not affect vesicle priming, but enhance or impair Ca2+-evoked release, respectively. The phenotypes caused by combining these mutations are dominated by the K603E and R769E mutations. Our results show that the C1-C2B region of Munc13-1 plays a central role in vesicle priming and support the notion that two distinct faces of this region control neurotransmitter release and short-term presynaptic plasticity.


Metabolism alters the selectivity of angiotensin-(1-7) receptor ligands for angiotensin receptors.
The present study examined whether metabolism of the putative angiotensin-(1-7) receptor agonist and antagonist [angiotensin-(1-7) and D-alanine(7) angiotensin-(1-7), respectively] altered their ability to interact with angiotensin AT(1), AT(2), and AT(4) receptor subtypes. Both angiotensin-(1-7) and D-alanine(7) angiotensin-(1-7) competed with low affinity for (125)I-sarcosine(1), isoleucine(8) angiotensin II binding to AT(1) and AT(2) receptors in rat liver and adrenal medulla membranes, respectively, and competed with low affinity for (125)I-angiotensin IV binding to AT(4) receptors in bovine kidney epithelial cell membranes. In vitro renal metabolism of the angiotensin-(1-7) receptor ligands (incubating peptides with rat cortical tissue homogenates) had minimal influence on low-affinity binding to AT(1) and AT(2) receptors, yet caused a significant and dramatic shift toward high-affinity binding for AT(4) receptors. Low-affinity angiotensin II binding to the AT(4) receptor was also shifted toward high-affinity binding following renal metabolism of the peptide. Conversely, angiotensins with high affinity for the AT(4) receptor (e.g., angiotensin IV) were shifted toward low-affinity binding states following peptide metabolism. Incubation of (125)I-angiotensin-(1-7) with rat cortical tissue generated the high-affinity AT(4) receptor ligand (125)I-angiotensin-(3-7), whereas the renal metabolism of (125)I-angiotensin II generated both (125)I-angiotensin-(3-7) and (125)I-angiotensin IV. These results reveal that renal metabolism of angiotensin-(1-7) receptor ligands and angiotensin II yields products that have high affinity for the AT(4) receptor and could potentially contribute to the biologic actions of the parent peptide in the kidney.


The role of climate-adaptive technological innovation in promoting agriculture carbon efficiency: impact and heterogeneity in economic development.
Achieving global climate change mitigation targets requires low-carbon production in agriculture. In such an endeavor, a new classification of climate-adaptive technology is defined to affect agriculture towards the low-carbon direction, but such an impact has seldom been empirically tested in the literature. In this paper, we investigate the impact of climate-adaptive technological innovation on agricultural carbon efficiency, a proxy for low-carbon agriculture. We use a stochastic directional distance function framework and a cross-country dataset covering 38 OECD countries. Additionally, we test the heterogeneous impact, considering that regional economic development is a crucial condition for deploying advanced technologies. The findings show that climate-adaptive technological innovation can promote carbon efficiency in agriculture, and this aggregate effect hides significant heterogeneity at different levels of economic development. The higher the economic development level is, the better climate-adaptive technological innovation contributes to improving agricultural carbon efficiency. Then, related policy implications are set forth.


Temporal lobe epilepsy with amygdala enlargement: A systematic review.
Temporal lobe epilepsy (TLE) with enlargement of the amygdala (AE) is a distinct clinical entity with contrasting clinical features from TLE with hippocampal sclerosis (HS). The objectives of this systematic analysis were to study the clinical characteristics and treatment outcome of people with TLE with AE. Pubmed, Embase, Cochrane, Web of Science, Scopus, and Medline were searched using the keywords amygdala enlargement, temporal lobe epilepsy, epilepsy, and seizure in November 2020. We found 18 studies that satisfied the inclusion criteria. A total of 361 patients were included in this analysis. The mean age of onset was 36.2 years, and febrile seizure was uncommon compared to TLE with HS subjects. The type of aura and automatism was similar to TLE with HS, though less prevalent. Electroencephalography (EEG) was usually concordant with the side of AE. Anti-seizure medications (ASM), surgical, and immunotherapy were used in different studies. 86 patients underwent surgery with Engel I outcome in 69.7%. Histopathology of the resected samples was predominantly dysplasia and gliosis. A group of patients that responded well to immunotherapy with subsequent reduction of amygdala volume (AMV) purported an autoimmune etiology of AE. Heterogeneity was the main drawback that prevented comparability among the studies. The methods of measurement of AMV also differed widely in the included studies, and standardization of its method is still lacking. This analysis suggests TLE with AE as a distinctive group of patients either due to a developmental anomaly or autoimmune etiology.


Valproic Acid versus Lamotrigine as First-line Monotherapy in Newly Diagnosed Idiopathic Generalized Tonic -Clonic Seizures in Adults - A Randomized Controlled Trial.
Idiopathic Generalized Tonic-Clonic Seizures (GTCS) are frequently encountered in adults. Their successful control is necessary to improve the quality of life of these patients. Valproic acid is a simple branched-chain carboxylic acid and lamotrigine is a phenyltriazine derivative. Opinions differ in regards to their effectiveness in idiopathic GTCS. To compare the effectiveness of valproic acid and lamotrigine in newly diagnosed adults with idiopathic generalized tonic-clonic seizures. The present prospective randomized study was conducted on 60 patients suffering from idiopathic GTCS. Thirty patients received valproic acid and rest 30 patients received lamotrigine. All patients were followed regularly monthly for one year for treatment response and adverse effects. After 12 months follow-up, 76.67% patients taking valproic acid and 56.67% patients taking lamotrigine were seizure-free. Common adverse effects recorded were nausea, dyspepsia, headache and skin rash. Valproic acid is more effective than lamotrigine as first-line drug in the treatment of adults with newly diagnosed idiopathic generalized tonic-clonic seizures.


Precise plant breeding using new genome editing techniques: opportunities, safety and regulation in the EU.
Several new plant breeding techniques (NPBTs) have been developed during the last decade, and make it possible to precisely perform genome modifications in plants. The major problem, other than technical aspects, is the vagueness of regulation concerning these new techniques. Since the definition of eight NPBTs by a European expert group in 2007, there has been an ongoing debate on whether the resulting plants and their products are covered by GMO legislation. Obviously, cover by GMO legislation would severely hamper the use of NPBT, because genetically modified plants must pass a costly and time-consuming GMO approval procedure in the EU. In this review, we compare some of the NPBTs defined by the EU expert group with classical breeding techniques and conventional transgenic plants. The list of NPBTs may be shortened (or extended) during the international discussion process initiated by the Organization for Economic Co-operation and Development. From the scientific point of view, it may be argued that plants developed by NPBTs are often indistinguishable from classically bred plants and are not expected to possess higher risks for health and the environment. In light of the debate on the future regulation of NPBTs and the accumulated evidence on the biosafety of genetically modified plants that have been commercialized and risk-assessed worldwide, it may be suggested that plants modified by crop genetic improvement technologies, including genetic modification, NPBTs or other future techniques, should be evaluated according to the new trait and the resulting end product rather than the technique used to create the new plant variety.


Effects of G6pc2 deletion on body weight and cholesterol in mice.
Genome-wide association study (GWAS) data have linked the G6PC2 gene to variations in fasting blood glucose (FBG). G6PC2 encodes an islet-specific glucose-6-phosphatase catalytic subunit that forms a substrate cycle with the beta cell glucose sensor glucokinase. This cycle modulates the glucose sensitivity of insulin secretion and hence FBG. GWAS data have not linked G6PC2 to variations in body weight but we previously reported that female C57BL/6J G6pc2-knockout (KO) mice were lighter than wild-type littermates on both a chow and high-fat diet. The purpose of this study was to compare the effects of G6pc2 deletion on FBG and body weight in both chow-fed and high-fat-fed mice on two other genetic backgrounds. FBG was reduced in G6pc2 KO mice largely independent of gender, genetic background or diet. In contrast, the effect of G6pc2 deletion on body weight was markedly influenced by these variables. Deletion of G6pc2 conferred a marked protection against diet-induced obesity in male mixed genetic background mice, whereas in 129SvEv mice deletion of G6pc2 had no effect on body weight. G6pc2 deletion also reduced plasma cholesterol levels in a manner dependent on gender, genetic background and diet. An association between G6PC2 and plasma cholesterol was also observed in humans through electronic health record-derived phenotype analyses. These observations suggest that the action of G6PC2 on FBG is largely independent of the influences of environment, modifier genes or epigenetic events, whereas the action of G6PC2 on body weight and cholesterol are influenced by unknown variables.


Mechanistic Insight into the Effect of Metal Ions on Photogeneration of Reactive Species from Dissolved Organic Matter.
The photogeneration of reactive species (RS) from dissolved organic matter (DOM) exhibits a great impact on the attenuation of pollutants in natural waters. However, the effect of metal ions on the photogeneration of excited triplet-state DOM (3DOM*), singlet oxygen (1O2), and hydroxyl radical (•OH) by effluent organic matter (EfOM), fulvic acid (FA), and humic acid (HA) is poorly understood. Here, we provided the first evidence that the quenching of 3DOM* was positively correlated with the complexation capacity of metal ions with DOM. Generally, the paramagnetic metal ions (Cr3+, Mn2+, Fe3+, and Cu2+) exhibited higher conditional stability constants (log KML) with DOM and stronger inhibition for RS than the others (Mg2+, Ca2+, Al3+, and Zn2+). For DOM of different sources, the metal binding capacity increased in the order of EfOM < HA < FA and the humic substances were more susceptible to metal ions. The inhibition was attributed to both static and dynamic quenching of 3DOM* by metal ions. The dynamic quenching rate constants of metal ions for 3DOM* were estimated as ∼109 M-1 s-1, which was positively related to the corresponding log KML. These findings highlight crucial links between metal-DOM complexation and 3DOM* quenching and, consequently, the inhibition of RS.


Egg perivitelline fluid proteome of a freshwater snail: Insight into the transition from aquatic to terrestrial egg deposition.
Proteins from the egg perivitelline fluid (PVF) are assumed to play critical roles in embryonic development, but for many groups of animals their identities remain unknown. Identifying egg PVF proteins is a critical step towards understanding their functions including their roles in evolutionary transition in habitats. We applied proteomic and transcriptomic analysis to investigate the PVF proteome of the eggs of Pomacea diffusa, an aerial ovipositing freshwater snail in the family Ampullariidae. The PVF proteins were separated with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method, and proteomic analysis was conducted using an LTQ Velos ion trap mass spectrometer coupled with liquid chromatography. Comparison of PVF proteomes and evolution analyses was performed between P. diffusa and other ampullariids. In total, 32 egg PVF proteins were identified from P. diffusa. They were categorized as PV1-like subunits, immune-responsive proteins, protein degradation, signaling and binding, transcription and translation, metabolism, oxidation-reduction and proteins with unknown function. Interestingly, the proteome includes a calcium-binding protein important in forming the hard eggshell that enabled the terrestrial transition. However, it does not include PV2, a neurotoxic protein that was assumed to be present in all Pomacea species. The PVF proteome data from P. diffusa can help us better understand the roles that reproductive proteins played during the transition from underwater to terrestrial egg deposition. Moreover, they could be useful in comparative studies of the terrestrialization in several groups of animals that occurred independently during their evolution.


Identification of the critical domains of the delta-opioid receptor involved in G protein coupling using site-specific synthetic peptides.
A large body of evidence implicates the second and third intracellular loops and the carboxyl-terminal portion of many G protein-coupled receptors as sites responsible for the interaction to G proteins. We synthesized a number of peptides from selected sites of the murine delta-opioid receptor and measured their ability to modify ligand-stimulated G protein activation and 3H agonist binding to the receptor. In membranes from Rat-1 fibroblasts transfected to express the murine delta-opioid receptor stably (clone D2 cells), the delta-opioid agonist [D-Ser2-Leu5-Thr6]enkephalin (DSLET) stimulated high affinity GTPase activity, which was inhibited by peptides that are derived from the proximal (i3.1) and the distal portions (i3.3) of the third intracellular loop with IC50 values of 15 +/- 5 and 50 +/- 4 microM, respectively. Peptides i3.1 and i3.3 inhibited DSLET-stimulated [35S]guanosine 5'-O-thiotriphosphate binding in the same membranes. However, a peptide designated i4, which was derived from a juxtamembranous region of the carboxyl-terminal tail of the delta-opioid receptor, failed to alter agonist-mediated high affinity GTPase activity or agonist-driven [35S]guanosine 5'-O-thiotriphosphate binding. Specific binding of [3H]DSLET to membrane preparations from clone D2 was reduced by peptides i3.1 and i4. Combinations of these peptides abolished detectable [3H]DSLET binding in the same membranes. Peptides i3.1 and i3.3 also destabilized the high affinity state of the receptor as assessed in 3H agonist binding on membranes from neuroblastoma X glioma (NG108-15) hybrid cells, which express the delta-opioid receptor endogenously; furthermore, delta-opioid receptor-stimulated GTPase activity in the same membranes was inhibited by peptides i3.1 and i3.3 but i4 was inactive. In contrast, peptides derived from the second intracellular loop (i2.1 and i2.2), an intermediate portion of the third intracellular loop (i3.2), and the extreme amino-terminal region of the receptor were without effect in these assays. These observations indicate that although peptides i3.1, i3.3, and i4 act via different mechanisms, they provide evidence that at least two sites of the third intracellular loop and part of the carboxyl-terminal tail of the delta-opioid receptor are important in the interaction between this receptor and cellular G proteins. Collectively, these results provide novel information about regions of the delta-opioid receptor that are involved in G protein coupling and high affinity agonist binding.


Recombinant human acid alpha-glucosidase enzyme therapy for infantile glycogen storage disease type II: results of a phase I/II clinical trial.
Infantile glycogen storage disease type II (GSD-II) is a fatal genetic muscle disorder caused by deficiency of acid alpha-glucosidase (GAA). The purpose of this study was to investigate the safety and efficacy of recombinant human GAA (rhGAA) enzyme therapy for this fatal disorder. The study was designed as a phase I/II, open-label, single-dose study of rhGAA infused intravenously twice weekly in three infants with infantile GSD-II. rhGAA used in this study was purified from genetically engineered Chinese hamster ovary (CHO) cells overproducing GAA. Adverse effects and efficacy of rhGAA upon cardiac, pulmonary, neurologic, and motor functions were evaluated during 1 year of the trial period. The primary end point assessed was heart failure-free survival at 1 year of age. This was based on historical control data that virtually all patients died of cardiac failure by 1 year of age. The results of more than 250 infusions showed that rhGAA was generally well tolerated. Steady decreases in heart size and maintenance of normal cardiac function for more than 1 year were observed in all three infants. These infants have well passed the critical age of 1 year (currently 16, 18, and 22 months old) and continue to have normal cardiac function. Improvements of skeletal muscle functions were also noted; one patient showed marked improvement and currently has normal muscle tone and strength as well as normal neurologic and Denver developmental evaluations. Muscle biopsies confirmed that dramatic reductions in glycogen accumulation had occurred after rhGAA treatment in this patient. This phase I/II first study of recombinant human GAA derived from CHO cells showed that rhGAA is capable of improving cardiac and skeletal muscle functions in infantile GSD-II patients. Further study will be needed to assess the overall potential of this therapy.


Robust adaptive fuzzy-neural control of nonlinear dynamical systems using generalized projection update law and variable structure controller.
In this paper, a robust adaptive fuzzy-neural control scheme for nonlinear dynamical systems is proposed to attenuate the effects caused by unmodeled dynamics, disturbance, and modeling errors. A generalized projection update law, which generalizes the projection algorithm modification and the switching-sigma adaptive law, is used to tune the adjustable parameters for preventing parameter drift and confining states of the system to the specified regions. Moreover, a variable structure control method is incorporated into the control law so that the derived controller is robust with respect to unmodeled dynamics, disturbances, and modeling errors. To demonstrate the effectiveness of the proposed method, several examples are illustrated in this paper.


Netrin-1 mediates neuronal survival through PIKE-L interaction with the dependence receptor UNC5B.
Netrins, a family of secreted molecules, have critical functions in axon guidance and cell migration during neuronal development. In addition to its role as a chemotropic molecule, netrin-1 also acts as a survival factor. Both UNC5 (that is, UNC5A, UNC5B, UNC5C or UNC5D) and DCC are transmembrane receptors for netrin-1 (Refs 8, 9). In the absence of netrin-1, DCC and UNC5 act as dependence receptors and trigger apoptosis. However, how netrin-1 suppresses the apoptotic activity of the receptors remains elusive. Here we show that netrin-1 induces interaction of UNC5B with the brain-specific GTPase PIKE-L. This interaction triggers the activation of PtdIns-3-OH kinase signalling, prevents UNC5B's pro-apoptotic activity and enhances neuronal survival. Moreover, this process relies strongly on Fyn because PIKE-L is tyrosine phosphorylated in response to netrin-1, and the netrin-1-mediated interaction of UNC5B with PIKE-L is inhibited in Fyn-null mice. Thus, PIKE-L acts as a downstream survival effector for netrin-1 through UNC5B in the nervous system.


Topical acne treatment with acetylcysteine: clinical and experimental effects.
Among the large variety of effects of acetylcysteine, several of them relate to cutaneous function in general and to sebaceous activity in particular. Due to the latter, a study was undertaken to investigate a possible effect of this substance in grade I acne. An analysis of the data from 99 patients in a double-blind study demonstrated that 5% acetylcysteine topical gel is significantly superior to placebo (P = .04) in reducing comedo counts. Comparable results were obtained in both sexes. This study indicates that acetylcysteine is an effective therapeutic option for the treatment of mild to moderate acne.


Influenza vaccine coverage, awareness, and beliefs regarding seasonal influenza vaccination among people aged 65 years and older in Central Saudi Arabia.
To investigate the level of awareness, sources of knowledge, and beliefs about the influenza vaccine in people ≥65 years in central Saudi Arabia. A cross-sectional study that used self-administered questionnaires conducted in 3 primary healthcare centers in Riyadh, Saudi Arabia between May 2018 and August 2018. There were 496 Saudis participants (70.2% male) were included, of whom, 47.8% had been vaccinated against influenza at least once. Of unvaccinated individuals, 46% believed that the vaccine was unnecessary. The majority of participants (70.2%) preferred to receive medical information on vaccination from doctors and health workers. Participants with higher levels of education were significantly more likely to have been vaccinated (p less than 0.05). Compared to the vaccinated participants, unvaccinated individuals were significantly less likely to be aware of the Ministry of Health campaign against influenza, believe that influenza vaccine does not weaken the immune system, know that elderly people and people with long-standing health problems should be regularly vaccinated against influenza, and believe that the influenza vaccine was the best way of prevention. Approximately 40% of the participants considered the influenza vaccine to be very effective and safe. There is a need for further programs in order to increase the awareness level of influenza vaccination. Also, health workers need to make more effort to educate patients because they are the preferred source of information.


Suicide in farmers in Scotland.
Farmers and farm workers have higher than expected rates of suicide and undetermined deaths in UK studies, and some rural areas of Scotland have higher than average male suicide rates. Firearm access seems to be an influencing factor in England and Wales. Type of farming, and farming social networks may also be important. This article describes suicide and undetermined deaths in male farmers and farm workers in Scotland from 1981-1999 using anonymised, routine data. Deaths of men aged 15-74 years from suicide or undetermined cause were identified from anonymised Scottish death records. Farmers and farm workers were identified using occupation codes. Methods of suicide used by farmers were compared with those of the general male population of the same age. A multiple linear regression was used to examine the influence of farm type, and the proportion of farmers in the working population of an area. 307 male farmers or farm workers died by suicide or undetermined cause in the time period. The overall rate was 31.4/100,000 per year (95% CI 28.1-35.1). Deaths using firearms were over-represented (29% of farming deaths compared with 3.6% in the general male population). There was no significant association between the male suicide rate in an area, and the farming suicide rate. Areas with lower proportions of farmers tended to have higher rates of farming suicide and undetermined deaths. This one factor described 85% of the variance among areas. Deaths were substantially more likely to involve firearms than suicide and undetermined deaths in the general male population. Less use of other methods did not completely compensate for this, indicating that method availability is likely to contribute to farming suicide rates. Farmers in areas where farming is less common were more likely to die by suicide, and this described most of the differences among areas. Networks and social supports may be important protective factors for farmers.


Hypoxemia after gastric bypass surgery for morbid obesity.
Fifty-six patients who underwent Roux-en-Y gastrojejunostomy for morbid obesity had arterial blood gas analysis before surgery and during the first five postoperative days. Preoperatively, seven subjects were hypoxemic and three were hypercapneic. Twenty-four hours after gastric bypass, 75% of the patients had an arterial oxygen pressure (Pao2) less than 60 mm Hg. Compared with preoperative measurements, blood gas values on the first postoperative day showed a 13.7-mm Hg decrease in the mean arterial Pao2 and a 5.0-mm Hg increase in the mean arterial carbon dioxide pressure. Arterial Pao2 determinations on the third, fourth, and fifth postoperative days returned toward, but remained significantly less than, paired values obtained before surgery. Patients with hypoxemia after gastrojejunostomy were significantly older and had significantly lower preoperative arterial Pao2 measurements than patients who were not hypoxemic after surgery. Weight, body mass index, and preoperative spirometric measurements did not distinguish between those patients who did and did not become hypoxemic postoperatively. We conclude that hypoxemia commonly follows gastric bypass for morbid obesity, and thus we recommend that all patients undergoing this procedure be treated with supplemental oxygen for at least the first three postoperative days.