[Unilateral approach for over the top bilateral lumbar decompression].
The main goal is bilateral microsurgical decompression of the cauda equina using a unilateral over the top approach. The challenge is to achieve decompression with minimal iatrogenic trauma to anatomical structures in the approach region and in the target area. Degenerative spinal disorders including lumbar central stenosis, lumbar lateral recess spinal stenosis, and foraminal narrowing. This technique is performed in patients presenting primarily with neurogenic claudication, leg or buttock symptoms, heaviness in the legs with or without radicular symptoms, with or without neurological deficits, and comparable MRI findings. There are no limitations regarding number of affected segments or the extent of narrowing. All available conservative treatment modalities not exhausted. Lack of serious neurological deficit. Minimally invasive, muscle-sparing and facet-joint-sparing bilateral enlargement of the lumbar spinal canal through a unilateral microsurgical cross-over approach. Patients are mobilized early 4-6 h postoperatively. Light sports activities (e.g., ergometer cycling, swimming) are allowed after 2 weeks. The same is true for the return to normal daily or work activities except for heavy physical work (usually 4 weeks out of work). Soft lumbar brace for 4 weeks (optional). The clinical outcomes are good to excellent. Meta-analyses and large case series report success rates for microsurgical decompression procedures of 73.5-95%. The reoperation rates are low (0.5-10%).


The role of structural biology in the design of sirtuin activators.
Sirtuins are NAD+-dependent protein lysine deacylases and mono-ADP-ribosylases whose activity regulates different pathways, including DNA damage repair, cell survival and metabolism, reactive oxygen species (ROS) detoxification, inflammation, cardiac function, and neuronal signaling. Considering the beneficial effects of specific sirtuin isoforms on health and lifespan, the past two decades have seen a mounting interest in the development of sirtuin activators. The availability of enzyme-activator co-crystal structures has proven significant throughout the years for elucidating the mechanisms of action of activators and designing more potent and selective molecules. In this review, we highlight the most interesting examples of sirtuin activators and provide comprehensive coverage of the role that structural biology played in their discovery and characterization.


Regulating Interfacial Lithium Ion by Artificial Protective Overlayers for High-Performance Lithium Metal Anodes.
The main limitation of lithium (Li) metal anodes lies in their severe dendrite growth due to nonuniform Li+ flux and sluggish Li+ transportation at the anode surface. Fabricating artificial protective overlayer with tunable surficial properties on Li metal is a precise and effective strategy to relieve this problem. In this Concept article, we focus on the basic principles of regulating interfacial Li+ through artificial protective overlayers and summarize the material preparation as well as structural design of these overlayers. The remaining challenges and promising directions of artificial protective overlayers are then highlighted to provide clues for the practical application of Li metal anodes.


Joint flexibility problems and the impact of its operationalisation.
Dissatisfaction is being voiced with the generally used way joint flexibility problems are defined (operationalised), i.e. as a range of motion (ROM) one or more degrees lower than normative ROM of healthy subjects. Other, specifically more function-related operationalisations have been proposed. The current study evaluated the effect of applying different operationalisations of joint flexibility problems on its prevalence. ROM data of 95 joints affected by burns of 23 children were used, and data on 18 functional activities (Burn Outcome Questionnaire (BOQ)). Five methods were used to operationalise joint flexibility problems: (1) ROM below normative ROM, (2) ROM below normative ROM minus 1SD, (3) ROM below normative ROM minus 2SD, (4) ROM below functional ROM, and (5) a score of 2 or more on the Likert Scale (BOQ). Prevalence of joint flexibility problems on a group level ranged from 13 to 100% depending on the operationalisation used. Per joint and movement direction, prevalence ranged from 40% to 100% (Method 1) and 0% to 80% (Methods 2-4). 18% of the children received '2' on the Likert Scale (Method 5). The operationalisation of joint flexibility problems substantially influences prevalence, both on group and joint level. Changing to a function-related operationalisation seems valuable; however, international consensus is required regarding its adoption. The study is registered in the National Academic Research and Collaborations Information System of the Netherlands (OND1348800).


Nano-SiO2 transport and retention in saturated porous medium: Influence of pH, ionic strength, and natural organics.
Nano silica (nSiO2), induces potential harmful effects on the living environment and human health. It is well established that SiO2 facilitates the co-transport of a variety of other contaminants, including heavy metals and pesticides. The current study focused on the systematic evaluation of the effects of multiple physicochemical parameters such as pH (5, 7, and 9), ionic strength (10, 50, and 100 mM), and humic acid (0.1, 1, and 10 mg/L) on the transport and retention of nSiO2 in saturated porous medium. Additionally, the influent concentration of nSiO2 (10, 50, and 100 mg/L) was also varied. Our experimental findings indicate that the size of nSiO2 aggregates was directly related to the pH, ionic strength, HA, and particle concentration had a significant impact on the breakthrough curves (BTCs). The stability provided by the varying concentrations of pH and humic acid had a significant effect on the size of nSiO2 aggregates and transport (C/C0 > 0.7). The presence of a greater magnitude of negative charge on the surface of both nSiO2 and quartz sand resulted in less aggregation and enhanced flow of nSiO2 through the sand column. The Electrostatic and steric repulsion forces were the primary governing mechanisms in relation to the size of nSiO2 aggregates, affecting the single-collector efficiency and attachment efficiency, which determined the maximal transport of nSiO2. Conversely, a probable increase in Van der Waals force of attraction exacerbated the particle deposition and reduced their mobility for high ionic strength, and particle concentrations (C/C0 < 0.1). The formation of large nSiO2 aggregates, in particular, was principally responsible for the enhancement of nSiO2 retention in sand columns over a broad range of IS and particle concentration. The interaction energy profiles based on the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory were determined to understand the mechanism of nSiO2 deposition. Aditionally, all the experimental BTCs were mathematically simulated and justified by the colloidal filtration theory (CFT). Considering the environmental ramifications, the transport behavior of nSiO2 was further evaluated in various natural matrices such as river, lake, ground, and tap water. The nSiO2 suspended in the river, lake, and tap water had significantly higher mobility (C/C0 > 0.7), whereas groundwater indicated higher retention (C/C0 < 0.3). The study advances our collective knowledge of physicochemical and environmental parameters that can affect particle mobility.


The prognostic significance of immunophenotype in high-grade non-Hodgkin's lymphoma.
The prognostic value of immunophenotyping lymphomas was assessed by studying 51 cases of high-grade non-Hodgkin's lymphoma for which long term clinical follow-up (14-28 years) was available. Using antibodies which identify T- and B-cell-related antigens in formalin-fixed, paraffin-embedded material, 43 were shown to be of B-cell and eight of T-cell phenotype. In terms of survival, cases of high-grade T-cell lymphoma fared significantly worse (P less than 0.05) than cases of high-grade B-cell subtype. These findings support the belief that T-cell lymphomas have a more aggressive clinical course than their B-cell counterparts.


Bayesian statistical inference enhances the interpretation of contemporary randomized controlled trials.
Randomized trials generally use "frequentist" statistics based on P-values and 95% confidence intervals. Frequentist methods have limitations that might be overcome, in part, by Bayesian inference. To illustrate these advantages, we re-analyzed randomized trials published in four general medical journals during 2004. We used Medline to identify randomized superiority trials with two parallel arms, individual-level randomization and dichotomous or time-to-event primary outcomes. Studies with P<0.05 in favor of the intervention were deemed "positive"; otherwise, they were "negative." We used several prior distributions and exact conjugate analyses to calculate Bayesian posterior probabilities for clinically relevant effects. Of 88 included studies, 39 were positive using a frequentist analysis. Although the Bayesian posterior probabilities of any benefit (relative risk or hazard ratio<1) were high in positive studies, these probabilities were lower and variable for larger benefits. The positive studies had only moderate probabilities for exceeding the effects that were assumed for calculating the sample size. By comparison, there were moderate probabilities of any benefit in negative studies. Bayesian and frequentist analyses complement each other when interpreting the results of randomized trials. Future reports of randomized trials should include both.


Three-month tapering and discontinuation of long- term, low-dose glucocorticoids in senior patients with rheumatoid arthritis is feasible and safe: placebo-controlled double blind tapering after the GLORIA trial.
The randomised placebo-controlled GLORIA (Glucocorticoid LOw-dose in RheumatoId Arthritis) trial evaluated the benefits and harms of prednisolone 5 mg/day added to standard care for 2 years in patients aged 65+ years with rheumatoid arthritis (RA). Here, we studied disease activity, flares and possible adrenal insufficiency after blinded withdrawal of study medication. Per protocol, patients successfully completing the 2-year trial period linearly tapered and stopped blinded study medication in 3 months. We compared changes in disease activity after taper between treatment groups (one-sided testing). Secondary outcomes (two-sided tests) comprised disease flares (DAS28 (Disease Activity Score 28 joints) increase >0.6, open-label glucocorticoids or disease-modifying antirheumatic drug (DMARD) increase/switch after week 4 of tapering) and symptoms/signs of adrenal insufficiency. In a subset of patients from 3 Dutch centres, cortisol and ACTH were measured in spot serum samples after tapering. 191 patients were eligible; 36 met treatment-related flare criteria and were only included in the flare analysis. Mean (SD) DAS28 change at follow-up: 0.2 (1.0) in the prednisolone group (n=76) vs 0.0 (1.2) in placebo (n=79). Adjusted for baseline, the between-group difference in DAS28 increase was 0.16 (95% confidence limit -0.06, p=0.12). Flares occurred in 45% of prednisolone patients compared with 33% in placebo, relative risk (RR) 1.37 (95% CI 0.95 to 1.98; p=0.12). We found no evidence for adrenal insufficiency. Tapering prednisolone moderately increases disease activity to the levels of the placebo group (mean still at low disease activity levels) and numerically increases the risk of flare without evidence for adrenal insufficiency. This suggests that withdrawal of low-dose prednisolone is feasible and safe after 2 years of administration.


Use of near-infrared spectroscopy combined with chemometrics for authentication and traceability of intact lemon fruits.
The reflectance NIR spectroscopy and chemometric data treatment on mature intact lemons, Limone di Sorrento PGI (cv Ovale di Sorrento) and Limone Costa D'Amalfi PGI (cv Sfusato Amalfitano) from Campania region, collected in 2018 and 2019, were used to predict properties, and discriminate cultivar and geographical provenance. By PCA, lemon NIR spectra grouped for production years due to the year variation of lemon properties attributable to annual climatic differences, homogeneous in all sites. This agrees with lemon chemical and physical differences by production year. Consequently, the relationship of NIR spectra with lemon quality properties by MLR and the cultivar and provenances discrimination by LDA were affected by year climatic difference; therefore, better model reliability was for single production year. NIR detectability of lemon properties did not appear beyond lemon thick peels, therefore the measured properties of lemon juices could derive from measurable properties of peel correlating with pulp properties.


[Cardiovascular surgery in patients with HIV: epidemiology, current indications, and long-term outcome].
The new highly active antiretroviral therapies used for the treatment of patients with human immunodeficiency virus (HIV) infection have dramatically modified the natural history of the disease. Although these therapies may themselves have adverse cardiovascular side-effects, their use has also resulted in seropositive patients surviving long enough to be exposed to the same risk factors as the general population. As a consequence, referrals for cardiovascular surgery have increased. We carried out a retrospective analysis of 39 consecutive HIV patients who underwent cardiovascular surgery between January 1998 and December 2006. The patients' mean age was 49+/-10 years and 77% (n=30) were male. The CD4-lymphocyte count was <500 cells/microl in 56% of patients (n=22). The main indications for surgery were endocarditis (n=18, 46%), coronary artery disease (n=12, 31%), and aortic disease (n=5, 13%). The in-hospital mortality rate was 5% (n=2). Postoperative complications included systemic infection (n=2, 5%), stroke (n=1, 3%), kidney failure (n=1, 3%) and respiratory failure (n=5, 13%). Repeat surgery for bleeding was necessary in 2 patients (5%). There were no postoperative complications in 29 patients (72%). After a clinical follow-up period of 8.2 years, the long-term mortality rate was 10.8% (4/37), giving survival rates at 1, 3 and 5 years of 91%, 86% and 81% respectively. Cardiovascular surgery can be performed with complete safety in HIV patients, and with promising results. This study indicates that, currently, long-term survival in this particular group of patients is potentially greater than 80%.


Spontaneous Medial Cribriform CSF Leak: Endoscopic Surgical Repair with Free Mucosal Graft-Our Experience.
Cribriform plate is the commonest site of spontaneous CSF leak, the fragility of the plate and juxtaposition of arachnoid's investment to the bone, where the olfactory nerve pierces the skull made this area, a vulnerable site for CSF leak. Transnasal endoscopic approach has gained popularity for CSF leak repair over the years. To describe the 5 year experience of spontaneous medial cribriform CSF leak repair with free mucosal graft in a tertiary medical centre. All patients who underwent transnasal endoscopic repair with free mucosal graft for spontaneous medial cribriform CSF leak in our institution between 2011 and 2016 were reviewed. Twelve patients were identified, all were women with a mean age of 44.5 years. The defect was localised by preoperative computed tomography scans with 1 mm cuts and MR cisternography. Via medial approach, the mucosa surrounding the entire defect was denuded and the defect was closed with free mucosal graft harvested either from the middle turbinate or from the nasal septum and middle turbinate was finally sutured with septum to stabilise the repair. The overall success rate was 100% with the first attempt with no recurrence or postoperative complications. Follow up ranged from 1 to 5 years. The endoscopic transnasal technique with free mucosal graft for the repair of spontaneous medial cribriform CSF rhinorrhoea is associated with a very high success rate and it should be considered for majority of cases.


Therapeutically useful mycobacteriophages BPs and Muddy require trehalose polyphleates.
Mycobacteriophages show promise as therapeutic agents for non-tuberculous mycobacterium infections. However, little is known about phage recognition of Mycobacterium cell surfaces or mechanisms of phage resistance. We show here that trehalose polyphleates (TPPs)-high-molecular-weight, surface-exposed glycolipids found in some mycobacterial species-are required for infection of Mycobacterium abscessus and Mycobacterium smegmatis by clinically useful phages BPs and Muddy. TPP loss leads to defects in adsorption and infection and confers resistance. Transposon mutagenesis shows that TPP disruption is the primary mechanism for phage resistance. Spontaneous phage resistance occurs through TPP loss by mutation, and some M. abscessus clinical isolates are naturally phage-insensitive due to TPP synthesis gene mutations. Both BPs and Muddy become TPP-independent through single amino acid substitutions in their tail spike proteins, and M. abscessus mutants resistant to TPP-independent phages reveal additional resistance mechanisms. Clinical use of BPs and Muddy TPP-independent mutants should preempt phage resistance caused by TPP loss.


Noninvasive measurement of cardiac pressures by induced ventricular wall resonance: preliminary results in the dog.
Experiments in the excised dog heart and in live dogs are presented wherein a method of induced right ventricular wall resonance has been utlized to estimate the pressure in the right ventricle of the heart. Externally induced cardiac wall vibrations produced low frequency signals which, when subjected to spectrum analysis and coherent signal processing, permitted close approximation of static pressures and wave forms in the right ventricle at normal and hypertensive levels.


Neurological symptoms in acute COVID-19 infected patients: A survey among Italian physicians.
COVID-19 is a pandemic disease and questions rise about the coronavirus 2 (Sars-CoV-2) effect on nervous system. This involvement could help explaining the pathogenesis of this condition and lead to novel therapeutic approaches. To assess the occurrence of neurological symptoms in COVID-19 patients during the Italian pandemic outbreak, as reported by physicians. In the early days of pandemic emergence we developed an online survey open to all Italian clinicians involved in the diagnosis and management of COVID-19 patients. The survey was structured in three sections, with nine different items concerning the presence of different specific clinical abnormalities. Each item was graded from "absent" to "severe" in a 4-point Likert's scale. Likert's scale data were analyzed by studying the distribution of responses by using medians and bar charts-relative frequencies. Also, in order to analyze differences in symptoms findings depending on the group of specialty, Likert's scale data were combined into two nominal categories ("absent"/"low" and "moderate"/"high"/") and a contingency table chi-square test was used. 126 physicians of 9 different medical specialties, from 10 regions of Italy, filled the online survey. The results show that 87.3% of practitioners reported neurological symptoms. In most cases these were mild and non-specific, but they were severe in a minority of patients. The most common symptoms observed were headache, myalgia and taste and smell abnormalities. Whilst there was no difference between neurologists and non-neurologists, we found that experienced clinicians (defined as clinicians that evaluated more than 30 patients) reported neurological symptoms more frequently than non-expert. Neurological symptoms have frequently been ported during the Italian COVID-19 pandemic, and thus should be monitored for all affected patients. Whilst some of the disturbances reported may be non-specific and common to other infectious diseases, smell and taste abnormalities might indicate nervous system as entry door for SARS-CoV-2 virus. This interpretation should promote research trials to avoid nervous system involvement.


Chronic application of curare does not increase the level of motoneuron survival-promoting activity in limb muscle extracts during the naturally occurring motoneuron cell death period.
Naturally occurring motoneuron cell death during development is a well-described phenomenon and the existence of a survival factor provided by target muscles has been postulated. Blockade of activity by chronic application of a neuromuscular junction blocker rescues almost all motoneurons from cell death. The present study was conducted in order to examine the possibility that the motoneuron survival-promoting activity in muscles increases following activity blockade. Cell culture was used to assess the degree of motoneuron survival-promoting activity present in muscle extracts. Embryonic chick motoneurons were labeled by injecting the water-insoluble fluorescent dye, DiI (Molecular Probes, Inc.) into the spinal nerves both before and during the cell death period. The labeled cells extending long neurites were counted after 2 days of culture as viable motoneurons in low-density heterogeneous cell cultures. The culture medium, Ham F12/DMEM (1:1 mixture) supplemented with 10% horse serum, 5% chick serum, and 5% fetal calf serum, was employed as a basic culture medium for assessing motoneuron survival factor, since it supported the survival of a significantly higher number of motoneurons derived from embryos before cell death than those during the cell death period, thus representing the motoneuron's requirement for survival factor in vivo. The number of surviving motoneurons clearly increased in proportion to the amount of muscle extract added to the culture medium. In comparison with control chick embryos, the dose-response relation between the number of surviving motoneurons and the amount of muscle extract added did not change when embryos were used after chronic application of curare. These results therefore indicate that survival factor derived from target muscle is crucial to the in vitro motoneurons during the cell death period, but do not support the idea that inactive muscle contains a higher amount of the survival factor.


Challenges and innovations of drug delivery in older age.
Both drug delivery performance and various age-related physical, mental and physiological changes can affect drug effectiveness and safety in elderly patients. The many drug delivery systems developed over the years include recent novel transdermal, nasal, pulmonary and orally disintegrating tablets that provide consistent, precise, timely and more targeted drug delivery. Certain drug delivery systems may be associated with suboptimal outcomes in the elderly because of the nature of drug present, a lack of appreciation of the impact of age-related changes in drug absorption, distribution and clearance, the limited availability of pharmacokinetic, safety and clinical data. Polypharmacy, patient morbidity and poor adherence can also contribute to sub-optimal drug delivery systems outcomes in the elderly. The development of drug delivery systems for the elderly is a poorly realised opportunity, with each system having specific advantages and limitations. A key challenge is to provide the innovation that best meets the specific physiological, psychological and multiple drug requirements of individual elderly patients.


Factors Associated with Low-Level Viremia in People Living with HIV in the Italian Antiviral Response Cohort Analysis Cohort: A Case-Control Study.
Despite effective antiretroviral therapies (ARTs), a subset of people living with HIV (PLWH) still experience low-level viremia (LLV, i.e., 50-1,000 copies/mL). The present study compared PLWH experiencing LLV with those maintaining virological suppression (VS) and explored the potential impact of preexisting drug resistance and other factors on LLV. We conducted a retrospective, 1:1 matched case-control study within a cohort of drug-experienced VS subjects from the Italian Antiviral Response Cohort Analysis database, followed in the period 2009-2019. Cases were individuals experiencing LLV, while controls were those who maintained VS. Matching was for calendar year of first ART regimen. Preexisting drug resistance was calculated as cumulative genotypic susceptibility score (GSS) according to regimen administered at the observational period start. To explore the effect of cumulative GSS, treated as a binary variable (≥2 and <2) and other factors on LLV, we performed a logistic regression analysis. Within a main population of 3,455 PLWH, 337 cases were selected. Cases were comparable to the controls for both gender and age. However, cases showed that they had experienced a longer time since HIV diagnosis, a higher number of drugs previously administered, lower baseline CD4+ T cell count and a higher zenith viral load (VL). By multivariate analysis, we found that higher zenith VL [adjusted odds ratio (aOR) (95% confidence interval [CI]) 1.30 (1.14-1.48)], a cumulative usage of both PI [aOR (95% CI): 2.03 (1.19-3.48)] and InSTI [aOR (95% CI): 2.23 (1.47-3.38)] and a cumulative GSS <2 [aOR (95% CI) 0.67 (0.46-0.98)], were associated with a higher risk in developing LLV. In current high-efficacy ART era, in drug-experienced PLWH, the predictors of increased risk of LLV were the presence of preexisting drug resistance, higher zenith VL, and previous PI, and InSTI exposure.


Leech predation on juvenile freshwater snails: effects of size, species and substrate.
The leechGlossiphonia complanata does not appear to have substantial impact on snail populations, but this may be due to most studies focusing on adult snails rather than juvenile snails. In this study I investigated how predation rates ofG. complanata feeding on newly-hatched and juvenile snails was affected by snail species, snail size, snail density and substrate, in a laboratory experiment. Number of snails eaten increased with increasing density resulting in a type II functional response curve. Predation rates were higher when leeches were feeding onLymnaea emarginata than onPhysa gyrina, whereas there was no significant difference in predation rates when they were feeding onL. emarginata andHelisoma anceps. Sandy substrates and greater snail size resulted in decreased predation rates. Sand reduced movement speed ofG. complanata, which probably reduced encounter rates. Thus, there was a comparatively large effect of leech predation on newly-hatched snails, due to a high probability of encounter and high predation rates, but spatial and temporal refuges probably reduce the importance of leech predation as a structuring force in freshwater snail assemblages.


Ischemia affects cardiac proteins in healthy animals less severely than in human patients.
Recently, our group showed that in human hearts proteins are extremely sensitive to ischemic injury. The purpose of this investigation was to evaluate the effects of ischemia on contractile and cytoskeletal proteins in rabbit and pig hearts and to compare these findings with those obtained in humans. Rabbit hearts were arrested by perfusion with Euro-Collins solution at different temperatures. Hearts perfused with buffer served as controls. Tissue samples were incubated for varying time intervals and processed for immunohistochemistry and electron microscopy. Porcine hearts were treated in the same manner. Changes in the localization of myosin, desmin, and tropomyosin antibodies were evaluated and the degree of ischemic injury was determined by electron microscopy. Healthy animal hearts tolerate ischemia better than human hearts. Cardiac proteins are more sensitive to ischemia than the ultrastructural cellular organelles. Temperatures as low as 0 degree C produce more cell damage than 4 degrees C and should therefore be avoided. The Euro-Collins solution protects the myocardium better than buffer. We conclude that healthy animal hearts are more resistant to ischemia than diseased human hearts and that results from experimental studies should be interpreted with caution with regard to the human situation.


An integrative model of shared decision making in medical encounters.
Given the fluidity with which the term shared decision making (SDM) is used in teaching, assessment and research, we conducted a focused and systematic review of articles that specifically address SDM to determine the range of conceptual definitions. In April 2005, we ran a Pubmed (Medline) search to identify articles published through 31 December 2003 with the words shared decision making in the title or abstract. The search yielded 681 citations, 342 of which were about SDM in the context of physician-patient encounters and published in English. We read and reviewed the full text of all 342 articles, and got any non-redundant references to SDM, which yielded an additional 76 articles. Of the 418 articles examined, 161 (38.5%) had a conceptual definition of SDM. We identified 31 separate concepts used to explicate SDM, but only "patient values/preferences" (67.1%) and "options" (50.9%) appeared in more than half the 161 definitions. Relatively few articles explicitly recognized and integrated previous work. Our review reveals that there is no shared definition of SDM. We propose a definition that integrates the extant literature base and outlines essential elements that must be present for patients and providers to engage in the process of SDM. The integrative definition of SDM is intended to provide a useful foundation for describing and operationalizing SDM in further research.


Stereoselective accumulation and biotransformation of chiral fungicide epoxiconazole and oxidative stress, detoxification, and endogenous metabolic disturbance in earthworm (Eisenia foetida).
>80 % of applied pesticides in agriculture will enter the soil and be exposed to soil animals. Little is known about the stereoselective metabolic effects of epoxiconazole (EPO) on soil animals. In this study, EPO-mediated stereoselective enrichment, biotransformation, oxidative stress, detoxification, and global metabolic profiles in earthworms were investigated by exposure to EPO and its enantiomers at 1 mg/kg and 10 mg/kg doses. Preferential enrichment of (-)-EPO was observed, and the five transformation products (TPs) exhibited the chemically specific stereoselective accumulation with inconsistent configurations. Biochemical markers related to reactive oxygen species (ROS) and detoxification (·OH- content, SOD, CAT, GST, and CYP450 enzymes) showed a significant stereoselective activation overall at the low-level exposure (p-value <0.05). Based on untargeted metabolomic analysis, the steroid biosynthesis and ROS-related biotransformation, glutathione metabolism, TCA cycle, amino acid metabolism, purine and pyrimidine metabolism of earthworms were significantly interfered with by EPO and its enantiomer exposure. More pronounced stereoselectivity was observed at the level of the global metabolic profile, while comparable levels of metabolic perturbations were identified at the individual metabolite level. This study provides novel insights into the stereoselective effects of the chiral fungicide EPO, and valuable evidence for soil environmental risk assessments.


Establishment of growth stimulating gene 2 protein time-resolved fluorescence immunoassay and its application in sepsis.
This study aimed to establish a highly sensitive time-resolved fluorescence immunoassay of growth stimulating express gene 2 protein (ST2-TRFIA) and evaluate its application value for sepsis. Two types of ST2 monoclonal specific antibodies against different epitopes of antigen molecule were used as coating and Eu3+-labeled antibodies. The double-antibody sandwich method was used in establishing ST2-TRFIA, and the methodology was evaluated. The established ST2-TRFIA was used in detecting ST2 concentration in the plasma samples of healthy controls and sepsis. The linear range of ST2-TRFIA was 1.446-500 ng/mL. Plasma ST2 concentrations detected through ST2-TRFIA were consistent with the results of fluorescence quantitative immunochromatography (ρ = 0.946). The plasma ST2 concentrations of patients with sepsis were significantly higher than those of healthy controls (P < 0.01). This study successfully established a highly sensitive ST2-TRFIA, which was highly comparable to commercially available fluorescent quantitative immunochromatographic kits and can facilitate the timely diagnosis of sepsis.


To-and-fro waveforms in the left gastric vein in portal hypertension.
Unusual to-and-fro waveforms were demonstrated in the left gastric vein on Doppler sonograms in four patients with liver cirrhosis. The patterns of the to-and-fro waveforms were diverse in each of the patients: both hepatopetal and hepatofugal flow occurred in a single waveform in case 1, changes in the flow direction with flow interruption were noted in case 2, and changes in flow direction without observation of flow interruption and changes after meals were noted in cases 3 and 4, respectively. These waveforms may represent a transitional phase during the development of a portal systemic shunt in patients with portal hypertension.


Optimization by hierarchical mutant production.
Inspired by the successful description of the first steps of molecular evolution by the quasispecies theory and the successful application of quasispecies-like algorithms to optimization problems, we propose a hierarchically organized algorithm. This new algorithm is able to solve a spin glass and a travelling salesman problem using only point mutations. Furthermore, it performs better under comparable circumstances than the ordinary quasispecies algorithm. Depending on the structure of the fitness landscape of the examined problem under consideration the hierarchically organized algorithm proves to be much more suitable than a simple quasispecies algorithm, especially in clustered landscapes. Tuning the error rates reveals the critical minimum copy fidelity necessary to guarantee optimization. We propose to incorporate hierarchical concepts into optimization algorithms inspired by biological evolution, such a genetic algorithms.


Psychosocial Mechanisms Underlying Older Black Men's Health.
To evaluate the psychosocial mechanisms underlying older Black men's self-rated health, we examined: (a) the individual, cumulative, and collective effects of stressors on health; (b) the direct effects of psychosocial resources on health; and (c) the stress-moderating effects of psychosocial resources. This study is based on a nationally representative sample of Black men aged 51-81 (N = 593) in the Health and Retirement Study (HRS). Ordinary least squares (OLS) regression models of the psychosocial determinants of self-rated health draw on data from the HRS 2010 and 2012 Core datasets and Psychosocial Modules. Each of the six measures of stressors as well as a cumulative measure of stressors are predictive of worse self-rated health. However, when considered collectively, only two stressors (chronic strains and traumatic events) have statistically significant effects. Furthermore, two of the five psychosocial resources examined (mastery and optimism) have statistically significant protective effects, and prayer moderates the harmful effects of traumatic events on self-rated health. Conventional measures of stressors and coping resources-originally developed to account for variance in health outcomes among predominantly white samples-may not capture psychosocial factors most salient for older Black men's health. Future research should incorporate psychosocial measures that reflect their unique experiences.


Self-regulation and early writing: A longitudinal examination from preschool through first grade.
Previous research has established the important role of self-regulation in supporting school readiness and academic learning. However, most of the research has not included examinations of the relation between early writing and self-regulation, although writing is an academic skill that also contributes to school readiness. Therefore, the purpose of this study was to explore the associations between self-regulation and early writing as children transitioned from preschool to first grade. Two groups of children were followed longitudinally. Group 1 (n = 171) was followed from preschool to kindergarten to Grade 1, and Group 2 (n = 196) was followed from a first year of preschool (Preschool 1) to a second year of preschool (Preschool 2) to kindergarten. Writing assessments included tasks at the word, sentence, and discourse levels. Self-regulation was measured using the Heads-Toes-Knees-Shoulders task. Writing and self-regulation were stable across the early years. Writing achievement was concurrently related to and significantly predicted later self-regulation. Self-regulation was concurrently associated with writing achievement but did not predict later writing achievement. Structural equation modeling results indicated that writing was a significant predictor of longitudinal change in self-regulation from preschool to kindergarten. The bidirectional relations between self-regulation and writing achievement were not supported by the data. Implications of the findings and future directions are discussed, including the effect of task demands in understanding relations between self-regulation and academic skills.


Impact of an embedded simulation team training programme in a paediatric intensive care unit: a prospective, single-centre, longitudinal study.
To evaluate the impact of an embedded simulation-based team training programme on perceived performance and to compare the effect over different phases of the programme. This was a prospective, single-centre, longitudinal study over the first 2 years of the programme. A total of 219 multidisciplinary health-care professionals participated in simulation sessions, followed by anonymous evaluation questionnaires. The programme was divided into three different phases: introductory (first 6 months), intermediate (second 6 months) and established phase (second year). A total of 88.7% of participants evaluated the impact on overall practice as effective, 56.5% reported a highly effective impact. A total of 90.9% (391/430) of questions on non-technical skills (communication and teamwork) showed an effective impact, 55.6% a highly effective impact, whereas only 70.2% (262/373) of questions on technical skills showed an effective impact. There was a significant (p < 0.001) increased score for effective impact in all categories between the introductory and intermediate phase, which was maintained throughout the established phase. Overall, 72.7% of the participants felt more confident to attend a future critical event, 32.5% were highly confident. In the longitudinal analysis there was a stepwise significant (p < 0.05) increase of confidence of participants. In a subgroup analysis (n = 143) there was a significant (p < 0.001) higher confidence in participants who had attended at least 3 sessions (90.7 vs. 61%). There is a 6- to 12-month learning curve in the implementation of an embedded multidisciplinary team training programme. Repeated exposure to simulation is most beneficial to crisis resource management training and single, isolated exposure may not be sufficient.


Primary tissue failure of the Freestyle stentless aortic root bioprosthesis.
We present a case of early tissue failure of the Medtronic Freestyle stentless bioprosthesis used in the aortic position which led to reoperation 27 months after implantation. Almost one half of the left coronary cusp of the prosthetic valve had torn away along the annulus. The tear started from the left-non coronary commissure and reached the midpoint of the base of the left coronary cusp. The prosthetic valve was easily excised and replaced with a new mechanical valve. There was no sign of calcification or vegetation in the excised valve. The cause of the valve dysfunction was thought to be primary tissue failure. Long-term function of the Freestyle stentless valve should be carefully followed up.


A study of in situ fluid bed melt granulation using response surface methodology.
The objective of this work was to investigate the influence of selected individual variables (binder content, inlet air temperature, and product endpoint temperature) of in situ fluid bed melt granulation on the granule particle size distribution and percentage of dissolved carvedilol using a three-factor, five-level circumscribed central composite design. Increased binder content had the effect of increasing the granule particle size and drug dissolution rate. The effect of inlet air temperature and product endpoint temperature was found to be more pronounced in case of granule particle size parameters. Within the studied intervals, the optimal quantity of binder as well as optimal process parameters were identified and validated using response surface methodology. Utilizing these optimal process and formulation parameters, successful scaling up of the fluid bed melt granulation process was carried out. Granule characteristics obtained at pilot scale are comparable to those obtained at laboratory scale.


Alterations in the ribosomal protein bL12 of E. coli affecting the initiation, elongation and termination of protein synthesis.
In bacteria, ribosomal protein bL12 forms the prominent stalk structure on the ribosome and binds to multiple, distinct translational GTPase factors during the sequential steps of translation. Using a genetic selection in E. coli for altered readthrough of UGA stop codons, we have isolated seven different mutations affecting the C-terminal domain of the protein that forms the interaction surface with translation factors. Analysis of these altered proteins, along with four additional alterations previously shown to affect IF2-ribosome interactions, indicates that multiple steps of translation are affected, consistent with bL12's interaction with multiple factors. Surprisingly, deletion of the release factor GTPase, RF3, has relatively little effect on bL12-promoted stop codon readthrough, suggesting that other steps in termination are also influenced by bL12.


Natural bioactives in perspective: The future of active packaging based on essential oils and plant extracts themselves and those complexed by cyclodextrins.
Aiming to attend to consumers' increasing demand for synthetic additives-free food products (stimulated by personal preferences or health concerns), the food industry has been trying to introduce natural bioactive compounds as food preservatives. In this respect, the development of active food packaging incorporated with natural compounds could be of great interest. However, this scenario still has some particularities that can hinder its application in the food industry, such as relatively reduced stability and, in some cases, undesirable sensorial properties. Cyclodextrins showed up as an option to circumvent these drawbacks by forming inclusion complexes that can protect the active compounds and perform their controlled release for contact with packaged food. For industrial manufacturing of active packages based on naturally occurring bioactive compounds, inclusion complexation can be considered one of the most promising alternatives. Therefore, this review summarizes the potential of nanotechnology in active food packaging by applying cyclodextrins as a carrier for natural active compounds, which can be provided by conventional sources and alternatively (and cost-effectively) agro-food by-products. The present study will benefit prospects for exploring cyclodextrins in active food packaging, which industrial application, without a doubt, will increase in the coming years.


Detection and classification of contrast-enhancing masses by a fully automatic computer-assisted diagnosis system for breast MRI.
To evaluate a fully automatic computer-assisted diagnosis (CAD) method for breast magnetic resonance imaging (MRI), which considered dynamic as well as morphologic parameters and linked those to descriptions laid down in the Breast Imaging Reporting and Data System (BI-RADS) MRI atlas. MR images of 108 patients with 141 histologically proven mass-like lesions (88 malignant, 53 benign) were included. The CAD system automatically performed the following processing steps: 3D nonrigid motion correction, detection of lesions by a segmentation algorithm, extraction of multiple dynamic and morphologic parameters, and classification of lesions. As one final result, the lesions were categorized by defining their probability of malignancy; this so-called morpho-dynamic index (MDI) ranged from 0%-100%. The results of the CAD system were correlated with histopathologic findings. The CAD system had a high detection rate of the histologically proven lesions, missing only two malignancies of invasive multifocal carcinomas and four benign lesions (three fibroadenomas, one atypical ductal hyperplasia). The 86 detected malignant lesions showed a mean MDI of 86.1% (± 15.4%); the mean MDI of the 49 coded benign lesions was 41.8% (± 22.0%; P < 0.001). Based on receiver-operating characteristic analysis, the diagnostic accuracy of the CAD system was 93.5%. Using an appropriate cutoff value (MDI 50%), sensitivity was 96.5% combined with specificity of 75.5%. The fully automatic CAD technique seems to reliably distinguish between benign and malignant mass-like breast tumors. Observer-independent CAD may be a promising additional tool for the interpretation of breast MRI in the clinical routine.


Response of plasma lipoproteins and acute phase proteins to myocardial infarction.
Plasma concentrations of lipoprotein-lipids, apolipoprotein B (apoB) and of seven other proteins have been estimated serially in 27 patients up to three months following myocardial infarction. Results were compared with those from age- and sex-matched control subjects. At three months the mean total, low density lipoprotein (LDL) and high density lipoprotein (HDL) cholesterol concentrations were higher than those of the control subjects, whereas very low density lipoprotein (VLDL) cholesterol, total and VLDL triglyceride, and total and LDL apolipoprotein B concentrations were not significantly different. Relative to concentrations at three months total and LDL cholesterol and apolipoprotein B concentrations fell markedly, and a slight fall occurred in HDL cholesterol following infarction. VLDL cholesterol and total and VLDL triglyceride were decreased only on day one. Albumin and transferrin concentrations were higher and alpha 1-acid glycoprotein was lower at three months than in the control subjects; alpha 2-macroglobulin, caeruloplasmin, haptoglobin and immunoglobulin IgM were not significantly different. Following infarction albumin and transferrin fell, alpha 2-macroglobulin did not change, and alpha 1-acid glycoprotein, caeruloplasmin, haptoglobin and IgM rose. The changes in both lipids and protein are probably part of the general metabolic response to trauma.


A New Black Fly Species of Simulium (Gomphostilbia) (Diptera: Simuliidae) From Thailand.
Simulium (Gomphostilbia) fukudae sp. nov. is described based on females, males, pupae, and larvae from Thailand. This new species is placed in the Simulium batoense species-group of Simulium (Gomphostilbia). It is characterized by the female frons almost bare, male paramere covered with minute setae, pupal gill composed of six slender filaments, pupal terminal hooks cone-shaped, and cocoon with an anterodorsal projection. Taxonomic notes are given to separate this new species from the most similar species, Simulium (G.) sextuplum Takaoka & Davies and Simulium (G.) paukatense Takaoka, both from Malaysia.


Apparent Intracellular Helicobacter pylori Detected by Immunohistochemistry: The Missing Link in Eradication Failure.
Helicobacter pylori is primarily an extracellularly living bacterium. However, seemingly intracellular occurrence can often be detected by immunohistochemical stains. Considering antimicrobial resistance, we investigated the impact of the apparent intracellular H. pylori (aiHp) on treatment failure of first-line triple therapies. Gastric biopsies of 814 patients infected with H. pylori naive to treatment were analyzed before and after eradication therapy by immunohistochemistry. Of these, 373 received treatment consisting of amoxicillin, clarithromycin, and proton pump inhibitor (AC/PPI). Availability of polymerase chain reaction-based clarithromycin susceptibility test results from pretreatment gastric biopsies was a precondition for matching 52 aiHp to 52 non-aiHp cases within the AC/PPI group. AiHp were detected mostly in low counts predominantly in corpus biopsies, rarely in antrum biopsies (95.2% vs 24.6%); they were found in 497 (61%) of all patients and in 192 of 373 patients (51.5%) in the AC/PPI group. The eradication rate in aiHp versus non-aiHp cases was 44.4% versus 72.9% in the entire sample and 45.3% versus 66.8% in the AC/PPI group. Among the 104 paired patients, respective values were 46.2% versus 78.8%; in clarithromycin-susceptible cases, 60.6% versus 91.9%. Both aiHp and resistance to clarithromycin proved to be highly significant (P ≤ .001) and independent predictors of eradication failure. Twelve of 13 aiHp cases with a clarithromycin-sensitive strain who failed eradication developed resistance to the antibiotic. AiHp found by immunohistochemical staining especially in corpus biopsies proved to be a risk factor for failure of first-line triple therapies; occurrence of aiHp should be considered with regard to therapy options.


Molecular organization of dengue fusion peptide in phospholipid monolayers revealed by tensiometry and vibrational spectroscopy.
The interaction of Dengue fusion peptide (FLAg) in selected lipid Langmuir monolayers was characterized with surface pressure-area isotherms and infrared spectroscopy to investigate the role of the membrane charge and molecular organization in the peptide-lipid binding. Surface pressure-area isotherms were employed to analyze the thermodynamic and mechanical properties of the FLAg-lipid monolayer, showing that charged lipid monolayers showed different peptide adsorption patterns for an optimized peptide concentration (maximum membrane adsorption). Polarization modulation infrared reflection-absorption spectroscopy pointed out that incorporating FLAg changed the dipole orientations for the lipid polar head groups, as confirmed in PG-containing monolayers. Also, FLAg reorients the lipid film when it interacts with the phosphate and choline groups. Finally, analysis of the 310-helix bands suggests that FLAg assumes a configuration as a hairpin, an essential premise for the beginning of the membrane fusion process.


Role of physician extenders: more regulation is necessary before full integration into practice.
To discuss the drawbacks and propose recommendations for integrating physician extenders in ophthalmologic practice. In this article, the role of utilizing physician extenders in ophthalmology is discussed. A role for physician extenders has been suggested as more and more patients will require ophthalmologic care. Guidance is needed on how to best integrate physician extenders into eye care. However, quality of care is of the highest importance, and unless there is reliable and consistent training of extenders, using physician extenders to administer invasive procedures (e.g., intravitreal injection) should be avoided due to safety concerns.


REIN: a tool to highlight inequalities in health
On the occasion of the 20th anniversary of the REIN (French Renal Epidemiology and Information Network), a summary work on the contributions of the national French ESKD register was carried out. On the issue of Social Inequalities in Health, the following key messages were retained. Social inequalities in health exist throughout the journey of a patient with chronic kidney disease and manifest as territorial inequalities in access to home-based or independent dialysis treatment and to transplant, whether preemptive or otherwise. SIH are observed in adults as well as in the paediatric population. The female gender appears to be associated with a disparity in access to kidney transplant.


Prevention of calpain-dependent degradation of STK38 by MEKK2-mediated phosphorylation.
Serine-threonine kinase 38 (STK38) is a member of the protein kinase A (PKA)/PKG/PKC-family implicated in the regulation of cell division and morphogenesis. However, the molecular mechanisms underlying STK38 stability remain largely unknown. Here, we show that treatment of cells with either heat or the calcium ionophore A23187 induced STK38 degradation. The calpain inhibitor calpeptin suppressed hyperthermia-induced degradation or the appearance of A23187-induced cleaved form of STK38. An in vitro cleavage assay was then used to demonstrate that calpain I directly cleaves STK38 at the proximal N-terminal region. Deletion of the N-terminal region of STK38 increased its stability against hyperthermia. We further demonstrated that the MAPKK kinase (MAP3K) MEKK2 prevented both heat- and calpain-induced cleavage of STK38. MEKK2 knockdown enhanced hyperthermia-induced degradation of STK38. We performed an in vitro MEKK2 assay and identified the key regulatory site in STK38 phosphorylated by MEKK2. Experiments with a phosphorylation-defective mutant demonstrated that phosphorylation of Ser 91 is important for STK38 stability, as the enzyme is susceptible to degradation by the calpain pathway unless this residue is phosphorylated. In summary, we demonstrated that STK38 is a calpain substrate and revealed a novel role of MEKK2 in the process of STK38 degradation by calpain.


Transanoproctoplasty: a 21-year review.
Transanal anorectoplasty was developed through the 1980s by the senior author (AB) as an alternative approach to posterior sagittal anorectoplasty for the management of imperforate anus. This study evaluates this surgical approach and its longer-term results. Case notes of all patients treated from 1984 to 2005 were reviewed. Operative procedures, colostomy requirement, complications, and long-term outcome were recorded. Patients were grouped according to the status of the pelvic floor, the location of the rectal fistula, and the sacral ratio. A total of 245 patients (175 male, 70 female) underwent transanal anorectoplasty. The perineum was well formed in 208 patients (85%), moderately formed in 15 (6%), and poorly formed in 22 (9%) patients. Two hundred three patients (82.8%) had a visible perineal or vulval fistula, 24 (9.8%) had a prostatic urethral fistula, whereas 18 (7.4%) had a bulbourethral fistula. Overall, 6 patients (2.5%) had wound infection or breakdown. As continence is age related, 32 patients were excluded from the study. Of the remaining 212 patients, 182 are continent with no soiling or only minimal staining. Thirty patients born with a poor perineum are incontinent. Transanal anorectoplasty is a safe procedure with limited morbidity. It is sphincter sparing and permits accurate placement of the rectum with its internal sphincter within the anal canal. The anus lies accurately placed at the center of the external anal sphincter muscle complex. Given a well-performed surgical intervention, eventual continence relates to the original anatomy and neurology of the pelvic floor. Transanal anorectoplasty should be regarded as an alternative rather than as a substitute to posterior sagittal anorectoplasty for reconstruction of most forms of imperforate anus.


The effects of gold nanoparticles in wound healing with antioxidant epigallocatechin gallate and α-lipoic acid.
Topical applications of antioxidant agents in cutaneous wounds have attracted much attention. Gold nanoparticles (AuNPs), epigallocatechin gallate (EGCG), and α-lipoic acid (ALA) were shown to have antioxidative effects and could be helpful in wound healing. Their effects in Hs68 and HaCaT cell proliferation and in mouse cutaneous wound healing were studied. Both the mixture of EGCG + ALA (EA) and AuNPs + EGCG + ALA (AuEA) significantly increased Hs68 and HaCaT proliferation and migration. Topical AuEA application accelerated wound healing on mouse skin. Immunoblotting of wound tissue showed significant increase of vascular endothelial cell growth factor and angiopoietin-1 protein expression, but no change of angiopoietin-2 or CD31 after 7 days. After AuEA treatment, CD68 protein expression decreased and Cu/Zn superoxide dismutase increased significantly in the wound area. In conclusion, AuEA significantly accelerated mouse cutaneous wound healing through anti-inflammatory and antioxidation effects. This study may support future studies using other antioxidant agents in the treatment of cutaneous wounds. In this study, topically applied gold nanoparticles with epigallocatechin gallate and alpha-lipoic acid were studied regarding their effects in wound healing in cell cultures. Significant acceleration was demonstrated in wound healing in a murine model.


Insurance Coverage of Biologics for Moderate-to-Severe Psoriasis: A Retrospective, Observational 5-Year Chart Review.
With the variability in health insurance coverage for psoriasis systemic therapies, recent changes in coverage for biologics have yet to be evaluated. To determine changes in insurance coverage of biologics for moderate-to-severe psoriasis between 2009 and 2014, with a focus on insurance policies as stated in prior authorization (PA) forms, coverage denials, and time course of approval process. A retrospective chart review was performed on patients with a diagnosis of psoriasis (International Classification of Diseases [ICD], Ninth Edition, code ICD 696.1) seen at the Department of Dermatology, Medical Faculty Associates, George Washington University between January 1, 2009 and December 31, 2014. Exclusion criteria included <9 % body surface area, loss to follow-up, lack of biologic treatment, biologic treatment via a clinical trial, and lack of health insurance. For all other patients, metrics collected included age, sex, body surface area, health insurance plan, prior therapies, prescribed biologic, PA necessity, time in days between PA submission and coverage decision, and denial justifications. Eight hundred and sixty-four patients with a diagnosis of psoriasis within the time period were identified, 114 of who met the inclusion criteria. PA requirement increased from 16 % of patients prescribed a biologic in 2009 to 75 % of patients prescribed a biologic in 2014. The mean duration in days between PA submission and coverage decision from the insurance company increased from 3.7 days in 2009 to 6.7 days in 2014. PA denial rates increased from 0 % in 2009 to 19 % in 2014. The most common reason for coverage denial was failure to attempt alternative therapies prior to requesting biologics. Insurance coverage of biologics for moderate-to-severe plaque psoriasis has become increasingly regulated between 2009 and 2014. Given both the cost burden and potential benefits of these therapies, further examination of healthcare coverage and treatment accessibility is warranted for optimal patient outcomes.


Collagenolytic activity in experimental intestinal anastomoses. Differences between small and large bowel and evidence for the presence of collagenase.
Collagen degradation is thought to be an integral part of the healing sequence of intestinal anastomoses, but almost nothing is known about the enzyme activities involved. We have studied collagenolytic activities, extracted from 1 day-old intestinal anastomoses in the rat. Using either soluble type I collagen or fibrillar type I or type III collagen as a substrate, activities measured in extracts from anastomotic segments were compared to those in extracts from uninjured intestine, removed at operation: in all cases, the collagenolytic activity in anastomotic extracts was significantly higher. This increase was significantly more pronounced in large bowel than in small bowel. The activities were strongly inhibited by serum and metallo-chelating compounds. Analysis, by means of SDS-polyacrylamide gel electrophoresis, of the reaction products of the degradation of fibrillar type I collagen by the extracts revealed the presence of a multitude of fragments, amongst them TcA fragments characteristic for the activity of mammalian collagenase. Thus, the degradative capacity towards various collagen substrates is enhanced in the anastomotic area during the first postoperative period and a true mammalian collagenase is one of the enzymes present.


Carbene-Catalyzed [4 + 2] Cycloadditions of Vinyl Enolate and (in Situ Generated) Imines for Enantioselective Synthesis of Quaternary α-Amino Phosphonates.
A carbene-catalyzed enantioselective addition of enals to five-membered cyclic imines is developed. The reaction gives chiral quaternary α-amino phosphonates bearing tetrasubstituted carbon centers with excellent enantioselectivities. The imine substrates can be generated in situ from the corresponding amines under an oxidative condition that is compatible with the carbene catalysis. Thus, a one-pot cross-dehydrogenative-coupling (CDC) reaction between enals and amines is also realized with high enantioselectivity remaining. The method provides quick enantioselective access to amino phosphonates with potential applications in medicines and pesticides.


Effects of cord milking in late preterm infants and full-term infants: A systematic review and meta-analysis.
Umbilical cord milking (UCM) consists of performing several milkings of the cord from the placenta to the newborn. The objective was to evaluate the effects of UCM on newborns ≥34 weeks' gestation. Searches were conducted in MEDLINE, EMBASE, CINAHL, the Cochrane Database of Clinical Trials, and the clinicaltrails.gov database for randomized clinical trials (RCT), with no time or language restrictions, and for articles that compared UCM with other strategies. The main results were initial hemoglobin and hemoglobin after 6 weeks. The data were collected by two reviewers and the quality of the studies was assessed using the Cochrane Manual methodology. The sample included 1845 newborns in 10 RCTs. The use of UCM in ≥34 weeks' gestation newborns was not related to initial hemoglobin levels (pooled weighted mean difference: (PWMD = 0.40 g/L [-0.16 to 0.95]) or after 6 weeks (PWMD = 0.07 g/L [-0.29 to 0.27]). A reduction in hemoglobin levels was also observed at 6 weeks when the control group had undergone late clamping (PWDM = 0.16g/L [-0.26 to -0.06]). UCM produced no differences in hematologic variables for newborns with ≥34 weeks of gestation relative to controls. However, a slight decrease in hemoglobin levels is observed at 6 weeks when the control group is made up of newborns with late clamping.


Ha-ras p21-GTP levels remain constant during primary keratinocyte differentiation.
Several lines of evidence that indicate that mutation of the Ha-ras oncogene is the initiating event in mouse skin carcinogenesis. Keratinocytes known to possess a mutated Ha-ras have been shown to be resistant to differentiation. Thus, overstimulation of the Ha-ras signaling pathway appears to block normal keratinocyte differentiation, and we hypothesized that for normal keratinocytes to terminally differentiate, the Ha-ras signaling cascade must be turned off. In the present studies, we measured the level and activity state of Ha-ras p21 protein in cultured keratinocytes undergoing calcium-induced differentiation. We have employed Western blot analysis to demonstrate that Ha-ras p21 protein levels remain constant during primary newborn and adult keratinocyte differentiation. The overall level of Ha-ras p21 was higher in immortalized, benign, and malignant mouse keratinocyte cell lines than in normal keratinocytes but did not change within each cell type when subjected to differentiating conditions. The percentage of Ha-ras p21 protein in its active, GTP-bound form also remained unchanged during primary adult keratinocyte differentiation and in immortalized, benign, and malignant keratinocytes subjected to differentiating conditions. Our results indicate that terminal differentiation of primary adult mouse keratinocytes occurred in the presence of constant levels of Ha-ras p21-GTP, suggesting that the Ha-ras signaling pathway may be blocked at a point distal to a step involving the Ha-ras p21 protein itself.


Dynamic responses to intravenous urapidil and dihydralazine in normal subjects.
Hemodynamic responses after urapidil were compared with those after dihydralazine in placebo-controlled, double-blind studies after cumulative intravenous doses. We recorded heart rate, blood pressure, systolic time intervals corrected for heart rate (electromechanical systole and preejection period), electrical impedance cardiography [(dZ/dt)/RZ index and mean electrical thorax impedance], and M-mode echocardiogram (end-systolic and -diastolic diameters, end-systolic wall stress, fractional shortening, and cardiac output). Both drugs induced dose-dependent reductions in total peripheral resistance, which resulted in reduction in left ventricular end-systolic wall stress and increases in heart rate (limited at +10 bpm with urapidil), fractional shortening, cardiac output, and the (dZ/dt)/RZ index. With each drug, diastolic blood pressure fell by 5 mm Hg, the corrected preejection period shortened (dihydralazine greater than urapidil), the corrected electromechanical systole did not change, and mean electrical thorax impedance rose with urapidil. The spectrum of effects indicates that both drugs reduce left ventricular afterload, thereby increasing left ventricular pump performance. Urapidil also exerts some preload reduction.


Nitric oxide generation is increased in experimental renal warm ischaemia-reperfusion injury.
Nitric oxide has a clearly defined place in normal renal homoeostasis while there is a continuing debate as to its role under pathophysiological conditions. This study investigated the role of nitric oxide in a model of renal warm ischaemia-reperfusion injury. Groups of rats underwent bilateral renal warm ischaemia (for 15-60 min) followed by reperfusion (20 or 80 min) before unilateral nephrectomy for measurement of renal nitric oxide (as nitroxides) and oxidative damage. Renal function was measured on days 2 and 7 before killing and nephrectomy. A further group received the nitric oxide synthase inhibitor N(G)-nitro L-arginine methyl ester (L-NAME; 50 mg per kg body-weight) before induction of warm ischaemia. In early reperfusion there was a correlation between the duration of warm ischaemia (15-45 min) and renal nitrate (r2=0.97) which increased from a mean(s.e.m.) baseline value of 95(5.9) to 208(17.3) nmol per mg protein following 45 min of warm ischaemia. Levels were further raised at 80 min and maintained through to day 7 (241(12.5) nmol per mg protein in 45-min group). This rise was attenuated by L-NAME (P< 0.01) as was the early rise in oxidative damage seen otherwise. By day 7, however, oxidative damage was increased (all P< or = 0.01). Renal nitric oxide increased early in recoverable warm ischaemia-reperfusion injury and remained raised to day 7. Nitric oxide synthase inhibition ameliorated early but exacerbated late damage suggesting that the early burst of nitric oxide is cytotoxic but that overall nitric oxide may exert a cytoprotective effect.


Identification of necroptosis-related gene TRAF5 as potential target of diagnosing atherosclerosis and assessing its stability.
Atherosclerosis (AS) is a leading cause of morbidity and mortality in older patients and features progressive formation of plaques in vascular tissues. With the progression of atherosclerosis, plaque rupture may occur and cause stroke, myocardial infarction, etc. Different forms of cell death promote the formation of a necrotic core of the plaque, leading to rupture. Necroptosis is a type of programmed cell death that contributes to the development of cardiovascular disease. However, the role of necroptosis in AS has not yet been investigated. The Gene Expression Omnibus (GEO) database was used to obtain gene expression profiles. Differentially expressed genes (DEGs) and necroptosis gene sets were used to identify necroptosis-related differentially expressed genes (NRDEGs). The NRDEGs were used to construct a diagnostic model and were further screened using least absolute shrinkage selection operator (LASSO) regression and random forest (RF) analysis. The discriminatory capacity of the NRDEGs was evaluated using receiver operating characteristic (ROC) curves. Immune infiltration levels were estimated based on CIBERSORTx analysis. The GSE21545 dataset, containing survival information, was used to determine prognosis-associated genes. Univariate and multivariate Cox regression analyses combined with survival analysis determined gene prognostic values. RNA and protein levels were detected by RT-qPCR and western blotting in arteriosclerosis obliterans(ASO) and normal vascular tissues. Vascular smooth muscle cells (VSMCs) were treated with oxidized low-density lipoprotein (ox-LDL) to develop cell models of advanced AS. The effects of protein knockdown on necroptosis were assessed by western blotting and flow cytometry. EdU and Cell Counting Kit-8 assays were used to examine cell proliferation. TNF Receptor Associated Factor 5 (TRAF5) was identified as a diagnostic marker for AS based on the AUC value in both the GSE20129 and GSE43292 datasets. According to differential expression analysis, LASSO regression analysis, RF analysis, univariate analysis, multivariate analysis, and gene-level survival analysis, TRAF5 was markedly associated with necroptosis in AS. Silencing TRAF5 promotes necroptosis and attenuates the proliferation of ox-LDL-induced cell models of advanced AS. This study identified a diagnostic marker of necroptosis-related atherosclerosis, TRAF5, which can also be used to diagnose and assess atherosclerotic plaque stability. This novel finding has important implications in the diagnosis and assessment of plaque stability in atherosclerosis.


Laboratory tools for oligo-secretory myeloma diagnosis: capillary electrophoresis versus free light chain assay.
To investigate how capillary electrophoresis (CE) works in oligo-secretory myeloma (OSM), we report a case here of OSM using multiple diagnostic methods including gel electrophoresis (GE), CE, and free light chain assay (sFLC). Also, we provide a brief review of laboratory methods to compare their diagnostic utilities in OSM. A 72 year-old Korean male suffering from low back pain during the past 6 months was transferred to the department of neurosurgery in order to evaluate abnormal findings in an imaging study, suggesting plasma cell myeloma (PCM) with multiple bone metastasis. CE showed no suspicious M-component; however, it showed increased Kappa components and skewing Kappa/Lambda ratio (K/L). Bone marrow examination revealed plasma cells observed up to 70%, which were compatible with sFLC results. Based on these results, the diagnosis turned out to be OSM with multiple bone metastases. Thereafter, the patient started the first cycle of chemotherapy accompanied by palliative radiation therapy. In our case, sFLC showed abnormal Kappa and K/L results from both serum and urine specimen. Therefore, it seems to be more sensitive and appropriate than both GE and CE to diagnose OSM.


Biochemical and structural insights into an Ochrobactrum sp. CSL1 ribose-5-phosphate isomerase A and its roles in isomerization of rare sugars.
Rare sugars have received increasing attention due to their important applications as sweeteners and building blocks. The substrate specificity and catalytic properties of ribose-5-phosphate isomerase A (RpiA) in isomerization of rare sugars have not been extensively explored. In this study, an RpiA from Ochrobactrum sp. CSL1 was cloned and expressed in Escherichia coli. The biochemical and reaction features were explored and its broad substrate specificity was identified. A higher reaction rate in isomerizing l-rhamnose to l-rhamnulose by OsRpiA, compared with OsRpiB found in the same strain indicated higher efficiency in preparing rare sugars, which was verified by kinetics study. The 2.8 Å resolution structure of OsRpiA was then solved and used in subsequent molecular dynamics experiments, providing a possible explanation for its distinct substrate specificity. The present study highlighted the unique role of microbial RpiA in preparing rare sugars, and its structural information provided a reliable reference for further reaction mechanism research and enzyme engineering work.


Establishment and characterization of a continuous murine uterine cervix cancer cell line metastatic to lymph nodes and lungs.
The murine uterine cervix cancer (MUCC) cell line was derived from a chemically induced Kunming mouse uterine cervix cancer (U27) and maintained in culture on solid substrates for over 100 passages. Cultures were morphotypically heterogeneous and heteroploid, with a modal number of chromosomes = 80. Each cell showed at least two abnormal chromosomes. Immunogold-silver staining was positive for keratin, vimentin, and laminin but not for desmin. The population doubling time was 27.8 h with a saturation density of 3.2 X 10(5) cells/cm2 and a peak mitotic index of about 6%. MUCC cells produced colonies on tissue culture plastic (68%) and in soft agar (8%). MUCC cells were fully malignant inasmuch as they produced in syngeneic mice invasive tumors that reproducibly were metastatic to lymph nodes and lungs. The MUCC cell line is the first mouse cervix cancer cell line useful for the study of invasion and metastasis.


The Learning Early About Peanut Allergy Study: The Benefits of Early Peanut Introduction, and a New Horizon in Fighting the Food Allergy Epidemic.
Observational studies have explored associations between timing of peanut, egg, and milk introduction and food allergy development, noting significant associations with reduced respective rates of milk, egg, and peanut allergy associated with earlier timing of introduction. Interventional studies developed to more definitively explore these outcomes have been published for egg and peanut, and are ongoing for multiple other allergens. This review focuses on the recent publication regarding the LEAP (Learning Early About Peanut Allergy) study, its highly favorable results, the policy implications of its findings, and the horizon for primary prevention as a realistic strategy to prevent food allergy.


Comparative Chemical Composition and Acetylcholinesterase (AChE) Inhibitory Potential of Cinnamomum camphora and Cinnamomum tamala.
Cinnamomum species have applications in the pharmaceutical and fragrance industry for wide biological and pharmaceutical activities. The present study investigates the chemical composition of the essential oils extracted from two species of Cinnamomum namely C. tamala and C. camphora. Chemical analysis showed E-cinnamyl acetate (56.14 %), E-cinnamaldehyde (20.15 %), and linalool (11.77 %) contributed as the major compounds of the 95.22 % of C. tamala leaves essential oil found rich in phenylpropanoids (76.96 %). C. camphora essential oil accounting for 93.57 % of the total oil composition was rich in 1,8-cineole (55.84 %), sabinene (14.37 %), and α-terpineol (10.49 %) making the oil abundant in oxygenated monoterpenes (70.63 %). Furthermore, the acetylcholinesterase inhibitory activity for both the essential oils was carried out using Ellman's colorimetric method. The acetylcholinesterase inhibitory potential at highest studied concentration of 1 mg/mL was observed to be 46.12±1.52 % for C. tamala and 53.61±2.66 % for C. camphora compared to the standard drug physostigmine (97.53±0.63 %) at 100 ng/ml. These multiple natural aromatic and fragrant characteristics with distinct chemical compositions offered by Cinnamon species provide varied benefits in the development of formulations that could be advantageous for the flavor and fragrance industry.


Inactivation of microsomal Ca(2+)-ATPase by 2-chloroethylethyl sulfide.
Exposure of liver microsomes to 2-chloroethylethyl sulfide (CEES) led to a dose-dependent decrease of Ca(2+)-ATPase activity. Studies on a structural requirement and a time dependence suggest that the enzyme inhibition may proceed via an instantaneous process involving an alkylation by an unstable intermediate, presumably a sulfonium form. It is noteworthy that the microsomal Ca(2+)-ATPase was more sensitive to CEES than the Na+/K(+)-ATPase from erythrocyte membranes. The Ca(2+)-ATPase was inhibited non-competitively by CEES, and its inhibitory action was independent of Ca2+ concentrations. The involvement of membrane phospholipid in the enzyme inhibition is excluded, since the temperature dependence of microsomal Ca(2+)-ATPase was not affected by CEES. Moreover, Triton X-100-solubilized Ca(2+)-ATPase was inactivated by the compound to the same extent as the membrane-bound enzyme was. Thus, it is suggested that CEES inactivates Ca(2+)-ATPase by alkylating the enzyme molecule at a region other than the active site.


Effect of starvation and elective surgery on hand dynamometry and circulating concentration of various proteins.
The effect of 4 days total starvation (water only) in five normal subjects on the circulating concentrations of various proteins was studied. Changes in plasma albumin and total protein concentrations were compared with those of six patients undergoing elective abdominal surgery with partial starvation and six patients undergoing orthopaedic surgery with adequate feeding - (0.126-0.146 MJ/kg/day and 1.2-1.4 g protein/kg/day). In a companion study hand grip strength was measured daily in ten normal subjects during starvation and in 18 patients undergoing surgery for hernia repair (n = 6), cholecystectomy (n = 6) and major abdominal surgery (n = 6). Starvation produced marked reductions (approximately 30%) in the circulating concentrations of retinol binding protein and prealbumin but did not significantly affect the plasma concentration of immunoglobulins (IgG, IgA, IgM) acute phase reactants (orosomucoid, haptoglobin, alpha(1) antitrypsin), albumin and total protein. On the other hand both types of elective surgery produced significant reductions in plasma albumin and total protein concentrations irrespective of feeding. Grip strength was not significantly altered by four days of starvation but surgery produced a temporary reduction in grip strength, the extent and duration of which was related to the severity of operation. This study helps to separate the effect of surgery and starvation on hand dynamometry and circulating protein concentrations and indicates their limitations as indicators of nutritional state.


Ketogenic diet: Assessing YouTube video information using quality, reliability, and text analytics methods.
Patients and the general audience refer social media platforms, such as YouTube, to learn and apply contemporary dietary methods. It is difficult for users to analyze the correctness and quality of information available on open platforms. Using scientific evaluation, this study assessed the quality, reliability, and content of YouTube videos on ketogenic diet (KD). Three experienced medical practitioners reviewed and evaluated 95 videos. The quality and reliability of the videos were assessed using the quality criteria for consumer health information and the global quality scale (GQS). Topic modeling and sentiment analysis were employed to determine the dominant themes and polarity of the information. Three types of publishers (doctors, educational institutions, and influencers) were identified for the study. The mean length of videos posted by doctors was high at 42.24 min. The reliability and quality scores ranging from 0 (low) to 5 (high) had an average of 3.08 ± 1.14 and 3.18 ± 1.18, respectively, for sampled videos. One-way analysis of variance reveals significant differences in DISCERN and GQS scores among doctors, educational institutions, and influencers. Topic discovery identified four themes: keto versus glucose, diabetes, KD food, and major chronic diseases. Sentiment analysis reveals positive content polarity, some content shared by doctors had a neutral sentiment. Content creators should augment the content by citing medical information and terminology. Viewers relied more on doctors for information related to KD. The aesthetic quality is high for all types of publishers. Publishers could focus on the discovered themes to create more content. Publishers should produce high-quality videos by improving esthetics (to increase engagement), and reliable medical information (to increase impact).


A description of Canadian epilepsy monitoring units: An initial step toward developing nursing practice consensus guidelines.
The epilepsy monitoring unit (EMU) is a well-established resource for investigating patients' seizures but is known to be heterogeneous in organization and clinical practice. The purpose of this study was to gain a better understanding of similarities and differences in EMU characteristics across Canada, with specific emphasis on EMU organization and nursing resources, which were currently unknown. Results would be used to develop a consensus on best nursing practice guidelines in EMUs with the goal to improve patient care and safety during epilepsy monitoring admissions. An 18-item survey was developed addressing EMU locations, types, nursing ratios, nursing roles, and other allied health resources. Surveys were distributed to lead nurses, physicians, and administrators in 29 EMUs across Canada. Results were tabulated and presented for each question in the survey. All EMUs were located in urban, teaching centers and divided similarly by patient age. The survey demonstrated considerable variability in EMU bed location and organization with the majority of EMUs being smaller, open units embedded in wards rather than larger, closed units. Independent of patient acuity, variability also existed in nurse-to-patient ratios, nursing skill level, specialty nursing support, and EEG technician availability. These findings highlight that EMU heterogeneity contributes to the challenges in the development of standardized safe care practices and that nursing education and nursing best practice recommendations need to be developed with baseline EMU nursing competencies, skills, and knowledge in mind.


Cerebrospinal fluid parameters and auditory brainstem responses following meningitis.
Auditory brainstem responses were measured in 94 children under 24 months of age immediately following treatment for bacterial meningitis. Evidence of peripheral hearing loss (thresholds of 30 dB HLn or greater) was found in 47% of the patients. In addition, 9% had prolonged interwave latencies, indicating the possible presence of retrocochlear pathology. Other clinical data were examined as well. CSF glucose concentration correlated with both the presence and magnitude of hearing loss (as measured by auditory brainstem responses). Magnitude of hearing loss also was associated with the presence of seizures. Although all children recovering from meningitis should be assessed for hearing loss, those who have had low CSF glucose concentrations and seizures appear to be at high risk.


Entrustable Professional Activities in Psychiatry: A Systematic Review.
Entrustable professional activities (EPAs) represent discrete clinical tasks that can be entrusted to trainees in psychiatry. They are increasingly being used as educational framework in several countries. However, the empirical evidence available has not been synthesized in the field of psychiatry. Therefore, the authors conducted a systematic review in order to summarize and evaluate the available evidence in the field of EPAs in undergraduate and graduate medical education in psychiatry. The authors searched PubMed, Cochrane Library, ERIC, Embase, PsycINFO, all Ovid journals, Scopus, Web of Science, MedEdPORTAL, and the archives of Academic Psychiatry for articles reporting quantitative and qualitative research as well as educational case reports on EPAs in undergraduate and graduate psychiatry education published since 2005. All included articles were assessed for content (development, implementation, and assessment of EPAs) and quality using the Quality Assessment Tool for Studies with Diverse Designs. The authors screened 2807 records and included a total of 20 articles in the final data extraction. Most studies were expert consensus reports (n = 6, 30%) and predominantly conducted in English-speaking countries (n = 17, 85%). Papers reported mainly EPA development and/or EPA implementation studies (n = 14, 70%), whereas EPA assessment studies were less frequent (n = 6, 30%). Publications per year showed an increasing trend both in quantity (from 1 in 2011 to 7 in 2018) and quality (from a QATSDD score of 27 in 2011 to an average score of 39 in 2018). The main focus of the articles was the development of individual EPAs for different levels of training for psychiatry or on curricular frameworks based on EPAs in psychiatry (n = 10, 50%). The lack of empirical controlled studies does currently not allow for meta-analyses of educational outcomes. The concept of EPA-based curricula seems to become increasingly present, a focus in the specialty of psychiatry both in UME and GME. The lack of empirical research in this context is an important limitation for educational practice recommendations. Currently there is only preliminary but promising data available for using EPAs with regard to educational outcomes. EPAs seem to be effectively used from a curriculum design perspective for UME and GME in psychiatry.


Massive subfascial hematoma after alteplase therapy for acute myocardial infarction.
The authors report a case of a patient who received alteplase for acute myocardial infarction and developed spontaneous subfascial hematoma without any evidence of direct trauma. Subfascial hematoma remains a rare and self-limited complication of thrombolytic therapy. The development of cutaneous ecchymosis associated with a sudden fall in hemoglobin after the administration of alteplase should strongly suggest the possibility of diffuse subfascial hematoma. Physicians should be aware of the possible association between the use of alteplase and the development of subfascial hemorrhage.


Structure determination and analysis of titin A-band fibronectin type III domains provides insights for disease-linked variants and protein oligomerisation.
Titin is the largest protein found in nature and spans half a sarcomere in vertebrate striated muscle. The protein has multiple functions, including in the organisation of the thick filament and acting as a molecular spring during the muscle contraction cycle. Missense variants in titin have been linked to both cardiac and skeletal myopathies. Titin is primarily composed of tandem repeats of immunoglobulin and fibronectin type III (Fn3) domains in a variety of repeat patterns; however, the vast majority of these domains have not had their high-resolution structure determined experimentally. Here, we present the crystal structures of seven wild type titin Fn3 domains and two harbouring rare missense variants reported in hypertrophic cardiomyopathy (HCM) patients. All domains present the typical Fn3 fold, with the domains harbouring variants reported in HCM patients retaining the wild-type conformation. The effect on domain folding and stability were assessed for five rare missense variants found in HCM patients: four caused thermal destabilization of between 7 and 13 °C and one prevented the folding of its domain. The structures also allowed us to locate the positions of residues whose mutations have been linked to congenital myopathies and rationalise how they convey their deleterious effects. We find no evidence of physiological homodimer formation, excluding one hypothesised mechanism as to how titin variants could exert pathological effects.


A small-molecule inhibitor targeting the mitotic spindle checkpoint impairs the growth of uterine leiomyosarcoma.
Uterine leiomyosarcoma (ULMS) is a poorly understood cancer with few effective treatments. This study explores the molecular events involved in ULMS with the goal of developing novel therapeutic strategies. Genome-wide transcriptional profiling, Western blotting, and real-time PCR were used to compare specimens of myometrium, leiomyoma, and leiomyosarcoma. Aurora A kinase was targeted in cell lines derived from metastatic ULMS using siRNA or MK-5108, a highly specific small-molecule inhibitor. An orthotopic model was used to evaluate the ability of MK-5108 to inhibit ULMS growth in vivo. We found that 26 of 50 gene products most overexpressed in ULMS regulate mitotic centrosome and spindle functions. These include UBE2C, Aurora A and B kinase, TPX2, and Polo-like kinase 1 (PLK1). Targeting Aurora A inhibited proliferation and induced apoptosis in LEIO285, LEIO505, and SK-LMS1, regardless of whether siRNA or MK-5108 was used. In vitro, MK-5108 did not consistently synergize with gemcitabine or docetaxel. Gavage of an orthotopic ULMS model with MK-5108 at 30 or 60 mg/kg decreased the number and size of tumor implants compared with sham-fed controls. Oral MK-5108 also decreased the rate of proliferation, increased intratumoral apoptosis, and increased expression of phospho-histone H3 in ULMS xenografts. Our results show that dysregulated centrosome function and spindle assembly are a robust feature of ULMS that can be targeted to slow its growth both in vitro and in vivo. These observations identify novel directions that can be potentially used to improve clinical outcomes for this disease.


The non-reversed vein femoro-distal bypass graft: a modification of the standard in situ technique.
The results of 85 in situ vein femoro-distal bypass grafts using a modified technique where the vein was completely mobilised but left "non-reversed" have been reviewed with particular regard to risk factors and complications. The distal anastomosis was to the infrageniculate popliteal artery in 55% and to the tibioperoneal trunk or a single calf vessel in the rest. Arteriographic run-off was by a single vessel in 42%. The primary failure rate at 1 month was 20% and the secondary failure rate 9%; the majority of early failures being due to missed technical errors despite the use of a pulse volume recorder. The cumulative secondary patency rate at 1 and 2 years was 77% and 72%; limb salvage 85% and 77%; and patient survival 89% and 83% respectively. The only significant risk factors were the level of the distal anastomosis and the run-off (P = 0.002 and 0.03 respectively). Complete mobilisation of the vein allows a tension free proximal anastomosis to the common femoral artery and avoids the risk of arteriovenous fistulae. A high vein utilisation rate of 93% was achieved by using a small 2.5 mm Hall valvulotome. Although there was a trend towards lower patency rates in veins with a minimum diameter less than 4 mm the results are still superior to PTFE. Compared to reversed vein the in situ technique has a better utilisation rate and the long-term patency rates are at least as good if not better. Improved methods of haemodynamic assessment during reconstruction to reduce technical errors may be the key to better early patency rates.


The potent mGlu receptor antagonist LY341495 identifies roles for both cloned and novel mGlu receptors in hippocampal synaptic plasticity.
Understanding the roles of metabotropic glutamate (mGlu) receptors has been severely hampered by the lack of potent antagonists. LY341495 (2S-2-amino-2-(1S,2S-2-carboxycyclopropyl-1-yl)-3-(xanth-9-y l)propanoic acid) has been shown to block group II mGlu receptors in low nanomolar concentrations (Kingston, A.E., Ornstein, P.L., Wright, R.A., Johnson, B.G., Mayne, N.G., Burnett, J.P., Belagaje, R., Wu, S., Schoepp, D.D., 1998. LY341495 is a nanomolar potent and selective antagonist at group II metabotropic glutamate receptors. Neuropharmacology 37, 1-12) but can be used in higher concentrations to block all hippocampal mGlu receptors, identified so far by molecular cloning (mGlu1-5,7,8). Here we have further characterised the mGlu receptor antagonist activity of LY341495 and have used this compound to investigate roles of mGlu receptors in hippocampal long-term potentiation (LTP) and long-term depression (LTD). LY341495 competitively antagonised DHPG-stimulated PI hydrolysis in AV12-664 cells expressing either human mGlu1 or mGlu5 receptors with Ki-values of 7.0 and 7.6 microM, respectively. When tested against 10 microM L-glutamate-stimulated Ca2+ mobilisation in rat mGlu5 expressing CHO cells, it produced substantial or complete block at a concentration of 100 microM. In rat hippocampal slices, LY341495 eliminated 30 microM DHPG-stimulated PI hydrolysis and 100 microM (1S,3R)-ACPD-inhibition of forskolin-stimulated cAMP formation at concentrations of 100 and 0.03 microM, respectively. In area CA1, it antagonised DHPG-mediated potentiation of NMDA-induced depolarisations and DHPG-induced long-lasting depression of AMPA receptor-mediated synaptic transmission. LY341495 also blocked NMDA receptor-independent depotentiation and setting of a molecular switch involved in the induction of LTP; effects which have previously been shown to be blocked by the mGlu receptor antagonist (S)-MCPG. These effects may therefore be due to activation of cloned mGlu receptors. In contrast, LY341495 did not affect NMDA receptor-dependent homosynaptic LTD; an effect which may therefore be independent of cloned mGlu receptors. Finally, LY341495 failed to antagonise NMDA receptor-dependent LTP and, in area CA3, NMDA receptor-independent, mossy fibre LTP. Since in the same inputs these forms of LTP were blocked by (S)-MCPG, a novel type of mGlu receptor may be involved in their induction.


DPPA3-HIF1α axis controls colorectal cancer chemoresistance by imposing a slow cell-cycle phenotype.
Tumor relapse is linked to rapid chemoresistance and represents a bottleneck for cancer therapy success. Engagement of a reduced proliferation state is a non-mutational mechanism exploited by cancer cells to bypass therapy-induced cell death. Through combining functional pulse-chase experiments in engineered cells and transcriptomic analyses, we identify DPPA3 as a master regulator of slow-cycling and chemoresistant phenotype in colorectal cancer (CRC). We find a vicious DPPA3-HIF1α feedback loop that downregulates FOXM1 expression via DNA methylation, thereby delaying cell-cycle progression. Moreover, downregulation of HIF1α partially restores a chemosensitive proliferative phenotype in DPPA3-overexpressing cancer cells. In cohorts of CRC patient samples, DPPA3 overexpression acts as a predictive biomarker of chemotherapeutic resistance that subsequently requires reduction in its expression to allow metastatic outgrowth. Our work demonstrates that slow-cycling cancer cells exploit a DPPA3/HIF1α axis to support tumor persistence under therapeutic stress and provides insights on the molecular regulation of disease progression.


Lu-atom-ordered oxonitridoaluminosilicate Ba0.9Ce0.1LuAl0.2Si3.8N6.9O0.1.
A single crystal of Ba0.9Ce0.1LuAl0.2Si3.8N6.9O0.1 (barium cerium lutetium aluminosilicate nitride oxide) was obtained by heating a mixed powder of Ba3N2, Si3N4, Al, Lu2O3, and CeO2 at 2173 K for 1 h under N2 gas at 0.85 MPa. X-ray single-crystal structure analysis revealed that the title oxynitride is hexa-gonal (lattice constants: a = 6.0378 (5) Å, c = 9.8133 (9) Å; space group: P63 mc) and isostructural with BaYbSi4N7. (Ba,Ce) and Lu atoms occupy twelvefold and sixfold coordination sites, respectively.


Progressive changes in lens crystallin glycation and high-molecular-weight aggregate formation leading to cataract development in streptozotocin-diabetic rats.
Because of their remarkable longevity, lens crystallins undergo a substantial amount of glycation (non-enzymatic glycosylation) during diabetic hyperglycemia. These post-translational modifications have the potential to disrupt the structural and functional properties of the lens crystallins and contribute to the formation of cataracts. Streptozotocin-induced diabetic rats were used to study the relationship between glycation of lens proteins and the formation of insoluble high-molecular-weight (HMW) aggregates believed to be responsible for cataract formation. After the onset of diabetes, cataracts developed in about 12- to 13 weeks. The animals were followed in this manner until cataracts developed and for an additional 63 days. Five control and five diabetic rats were killed every 3 weeks and lenses removed. Levels of glycated protein and glycated amino acids in lenses from each animal were examined by affinity chromatography. In addition, the changes in crystallin composition and development of HMW aggregates were monitored by molecular-sieve HPLC techniques. As diabetic hyperglycemia continued there was a linear increase in glycated protein in both the soluble and insoluble fractions. This increase was paralleled by an increase in the soluble HMW and insoluble HMW aggregates. Other changes included a decrease in reactive sulfhydryls which indicates an increase in disulfide bond formation. The gamma-crystallin levels also decreased in a linear fashion during the hyperglycemic pre-cataract and cataract stages. It appears that the glycation of lens crystallin, the disappearance of reactive sulfhydryls and the formation of HMW aggregates are interrelated.


[Enlargement labial commissuroplasty using the Préaux-Texier-Réal method. Our personal experience].
The augmentation labial commissuroplasty technique, as described by Préaux, Texier and Réal, after reduction of the buccal opening by lip-to-lip autoplasty is presented. The results achieved, the feasibility and the advantages of this procedure are compared with those of traditional techniques.