Protein S-nitrosylation in photosynthetic organisms: A comprehensive overview with future perspectives.

BACKGROUND: The free radical nitric oxide (NO) and derivative reactive nitrogen species (RNS) play essential roles in cellular redox regulation mainly through protein S-nitrosylation, a redox post-translational modification in which specific cysteines are converted to nitrosothiols. SCOPE OF VIEW: This review aims to discuss the current state of knowledge, as well as future perspectives, regarding protein S-nitrosylation in photosynthetic organisms. MAJOR CONCLUSIONS: NO, synthesized by plants from different sources (nitrite, arginine), provides directly or indirectly the nitroso moiety of nitrosothiols. Biosynthesis, reactivity and scavenging systems of NO/RNS, determine the NO-based signaling including the rate of protein nitrosylation. Denitrosylation reactions compete with nitrosylation in setting the levels of nitrosylated proteins in vivo. GENERAL SIGNIFICANCE: Based on a combination of proteomic, biochemical and genetic approaches, protein nitrosylation is emerging as a pervasive player in cell signaling networks. Specificity of protein nitrosylation and integration among different post-translational modifications are among the major challenges for future experimental studies in the redox biology field. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock.

Predicting the functional states of human iPSC-derived neurons with single-cell RNA-seq and electrophysiology.

Human neural progenitors derived from pluripotent stem cells develop into electrophysiologically active neurons at heterogeneous rates, which can confound disease-relevant discoveries in neurology and psychiatry. By combining patch clamping, morphological and transcriptome analysis on single-human neurons in vitro, we defined a continuum of poor to highly functional electrophysiological states of differentiated neurons. The strong correlations between action potentials, synaptic activity, dendritic complexity and gene expression highlight the importance of methods for isolating functionally comparable neurons for in vitro investigations of brain disorders. Although whole-cell electrophysiology is the gold standard for functional evaluation, it often lacks the scalability required for disease modeling studies. Here, we demonstrate a multimodal machine-learning strategy to identify new molecular features that predict the physiological states of single neurons, independently of the time spent in vitro. As further proof of concept, we selected one of the potential neurophysiological biomarkers identified in this study-GDAP1L1-to isolate highly functional live human neurons in vitro.

[Symptomatic lumbar incisional hernia after open nephrectomy: What are the risk factors?]. / Éventrations lombaires symptomatiques après néphrectomies par lombotomie: quels sont les facteurs de risque?

OBJECTIVE: Lumbar incisional hernias after open nephrectomy are rare but can lead to aesthetic disorder, discomfort or intestinal obstruction. The aim of the study is to highlight their risk factors. PATIENTS AND METHODS: The characteristics of patients who suffered from symptomatic and surgically treated lumbar incisionnal hernia after open nephrectomy (study group "GE") were compared to those of patients who underwent open nephrectomy without postoperative incisional hernia (control group "GT") using the Student's t test and Mann-Whitney test (statistical significance P value<0.05). GT patients were randomly selected with a 1/4 ratio (1 lumbar incisionnal hernia vs 4 controls). RESULTS: From 2004 to 2014, 417 open nephrectomies were performed in one university hospital. Forty-five patients were included: 9 in GE and 36 in GT. There was no statistically significant difference between GT and GE for weight, height, body mass index (BMI), emergency, partial nephrectomy, laterality, rib resection, laparoscopic conversion to open surgery, postoperative complications, smoking, diabetes mellitus, cardiovascular history, obesity and sex, but there was a statistically significant difference for age, operative time, chronic obstructive pulmonary disease (COPD) and severe obesity (BMI>35) with, respectively, P=0.05, P=0.02, P=0.04 and P=0.02. CONCLUSION: Risk factors for lumbar incisional hernia after open nephrectomy are age, operative time, severe obesity and COPD. LEVEL OF EVIDENCE: 5.

Study of the impacts of patient-educators on the course of basic sciences in dental studies.

Ever since 2006, Nantes University dental educators have started organising lectures led by the mother of a young patient suffering from ectodermic dysplasia (patient-educator) to help second-year students to better understand how important it is for their future dental work to better understand basic sciences. In this study, we have analysed this training experience on students' motivation. For this purpose, students were asked to complete questionnaires 10 days after the patient-educator's lecture (early assessment; n = 193) and 4 years later, during the last year of their dental studies (delayed assessment; n = 47). Moreover, 3 years after the first lecture, we analysed the ability of students to diagnose a mother carrying the ectodermic dysplasia genetic disorder, using a case-based learning exercise with a patient showing dental features similar to those exposed by the patient-educator (measure of knowledge; n = 42). Ten days after the lecture, the early assessment shows that all the students were interested in the lecture and 59% of the students declared being motivated to find out more about genetics whilst 54% declared the same thing about embryology courses. Moreover, 4 years later, 67% of the students remembered the patient-educator's lecture a little or very well. Three years after the course, 83% of the students diagnosed ectodermal dysplasia whilst studying the case-based example that listed typical dental phenotypes. In conclusion, this study shows that this original educational approach enhances dental students' motivation in learning basic sciences and that patient-educators could offer many benefits for students and patients.

[Is obesity a barrier to kidney transplantation?]. / L'obésité est-elle un frein à la transplantation rénale ?

INTRODUCTION: Kidney transplantation is the most suitable of ESRD care. The proportion of obese people is increasing in the general population and patients with kidney impairment. It is important to assess the impact of obesity on surgical complications of kidney transplantation. The aim of this retrospective study was to signify the correlation between obesity and the occurrence of postoperative urological complications during the first year. METHODS: We conducted a retrospective study from March 1999 to December 2009. We conducted a chart review of patients undergoing kidney transplantation. The kidneys were taken from cadaveric donors. Data collected included age, weight, height, preoperative BMI; causal nephropathy, smoking, hypertension, diabetes, anticoagulation therapy. Intraoperative data included operative time (DO), cold ischemia. Urological complications were recorded during the first year after the kidney transplantation (vascular anastomotic strictures, ureterovesical stenosis, lymphorrheas, pyelonephritis, hematoma, wound infection). Statistical analysis consisted of a t-test for independent samples and univariate and multivariate logistic regression for the occurrence of complications. RESULTS: Four hundred and twenty-two patients were transplanted in total. We excluded 20 patients. BMI and duration of surgery patients with complications were significantly different from those of patients with no complications (P=0.016 and P=0.039, respectively). Obese (n=48) had more diabetes (12.5% versus 3.7%, P=0.014), were more often smoking (35.4% versus 22%, P=0.012), had a longer DO (203.64minutes versus 182.46minutes, P=0.006), and complications (62.5% versus 50.28%, P=0.03) than patients with a BMI <30 (n=354). After adjusting for age, smoking, DO, diabetes and BMI showed that only BMI was an independent predictor of the occurrence of postoperative complications with P=0.048 and RR=1.058 [CI: 1 to 1.119]. However, there was no more transplantectomy obese (P=0.911). CONCLUSION: Our study showed that there is a significant risk of surgical complications after kidney transplantation in obese patients. But ultimately, this does not affect graft survival because there are no more transplantectomies or return to dialysis. LEVEL OF EVIDENCE: 5.

Poor long-term stability of the Corvita abdominal stentgraft.

INTRODUCTION: The endovascular treatment of abdominal aortic aneurysms was introduced in the early 1990s, with different generations of devices using various options for either the stent skeleton or the membrane. REPORT: Corvita generated one of these devices using braided stainless steel and a porous spun polycarbonate urethane membrane. DISCUSSION: In this report, we describe a case involving Corvita stentgraft explantation for complete aneurysm reperfusion after 13 years, demonstrating major degradation of the polyurethane membrane.

[The impact of obesity on renal function at one year after kidney transplantation: single-center experience]. / Impact de l'obésité sur la fonction rénale à 1 an chez les transplantés rénaux: expérience mono centrique.

INTRODUCTION: Kidney transplantation is the treatment of choice for ESRD. Several studies have investigated the factors that may affect kidney function at 1 year. The factors mentioned are anemia, hypercholesterolemia, immunosuppressors, etc. We studied the independent predictors of serum creatinine>100µmol/L at 1 year. MATERIALS AND METHODS: We conducted a retrospective study from March 1999 to December 2009. We conducted a chart review of 402 kidney transplant patients. The kidneys were removed from cadaveric donors. Data collected included age, weight, height, preoperative BMI, the causal nephropathy, smoking, hypertension, diabetes, anticoagulation. Intraoperative data included operative time, and cold ischemia. Statistical analysis consisted of a t-test for independent samples comparing the group with a creatinine≤100µmol/L vs>100 group, and univariate and multivariate Cox regression for a serum creatinine>100µmol/L at 1 year and test of correlation between BMI and serum creatinine at 1 year postoperatively. RESULTS: We found a significant difference in BMI and cold ischemia with P=0.008 and P=0.002, respectively. In contrast there was no difference in age, operative time and blood loss, P=0.758, P=0.941 and P=0.963, respectively. Multivariate Cox regression showed that donor age P=0.004 (HR: 1.016 and CI: 1.005-1.027), a recipient age P=0.023 (HR: 0.986 and CI: 0.974-0.998) and BMI P=0.001 (HR: 1.019 and CI: 1.010-1.028) were independent predictors of serum creatinine>100µmol/L at 1 year. The Pearson correlation coefficient r=0.154 (P=0.004) showed a significant correlation between BMI and serum creatinine. CONCLUSION: Our study showed that donor age, recipient age and BMI were independent predictors of renal function>100µmol/L at 1 year. Our results highlight the difficulty of the management of obesity in renal transplant patients. LEVEL OF EVIDENCE: 5.

D-cycloserine or similar physiochemical compounds may be uniquely suited for use in Bacillus anthracis spore decontamination strategies.

AIMS: As observed in the aftermath of the anthrax attacks of 2001, decontamination and remediation of a site contaminated by the accidental or intentional release of Bacillus anthracis spores is difficult, costly and potentially damaging to the environment. The identification of novel strategies that neutralize the threat of spores while minimizing environmental damage remains a high priority. We investigated the efficacy of d-cycloserine (DCS), an antibiotic and inhibitor of the spore-associated enzyme (alanine racemase) responsible for converting l-alanine to d-alanine, as a spore germination enhancer and antimicrobial agent. METHODS AND RESULTS: We characterized the impact of DCS exposure on both germinating spores and vegetative cells of fully virulent B. anthracis by evaluating spore germination kinetics, determining the minimum inhibitory concentrations (MICs) required to affect growth of the bacteria and performing macrophage viability assays. DCS enhanced germination induced by l-alanine and also efficiently killed the newly germinated spores. Furthermore, DCS proved nontoxic to macrophages at concentrations that provided protection from the killing effects of spores. Similar tests were conducted with Bacillus thuringiensis (subspecies kurstaki and Al Hakam) to determine its potential as a possible surrogate for B. anthracis field trials. Bacillus thuringiensis spores responded in a similar manner to B. anthracis spores when exposed to DCS. CONCLUSIONS: These results further support that DCS augments the germination response of spores in the presence of l-alanine but also reveal that DCS is bactericidal towards germinating spores. SIGNIFICANCE AND IMPACT OF THE STUDY: DCS (or similar compounds) may be uniquely suited for use as part of decontamination strategies by augmenting the induction of spore germination and then rendering the germinated spores nonviable.

Impact of routine completion angiography on the results of primary carotid endarterectomy: a prospective study in a teaching hospital.

OBJECTIVE: To assess the usefulness of completion angiography in the prevention of stroke, carotid occlusion and residual stenosis after primary carotid endarterectomy (CEA) in the setting of a teaching hospital. MATERIAL AND METHODS: From January 1995 to August 2009, 1055 consecutive patients having 1179 CEAs were entered in a prospective study excluding patients with severe renal insufficiency, allergy to contrast media and patients with repeat CEA or carotid bypass. In this cohort, 552 patients (52.3%) were asymptomatic, 318 (30.2%) had a transient ischaemic attack (TIA) and 185 (17.5%) had a stroke. Routine completion angiography was obtained in all 1055 patients. The decision to perform a surgical revision was decided for any of the following defects: (1) a residual stenosis of more than 50% of the internal carotid artery (ICA) or common carotid artery (CCA) and of more than 70% of the external carotid artery (ECA), (2) any flap and (3) any intraluminal-filling defect. A postoperative duplex scan was obtained within a week after surgery and thereafter on a yearly basis. Median follow-up was 7 years. RESULTS: CEA was performed by a senior surgeon as first operator in 812 cases (69%) and by a trainee, with a scrubbed senior surgeon, in 367 cases (31%). Completion angiography revealed significant defects in 72 cases (6.1%) warranting revision for ECA flap (n = 30), thrombus in contact with the patch (n = 7), distal ICA flap or stenosis (n = 20) and CCA flap or residual plaque (n = 15). Logistic regression analysis showed that total length of the carotid plaque >6 cm (p = 0.02, Odds ratio: 2.31; 95% confidence interval (CI) (1.21-3.72)), eversion endarterectomy of the ECA (p = 0.01, Odds ratio 3.41; 95%CI (2.10-5.94)) and trainee as first operator (p = 0.02, Odds ratio 2.42; 95%CI (1.81-4.23)) were independent predictors of operative defects seen on completion angiography. No complication in relation to carotid catheterisation or injection of contrast media occurred in this series. The 30-day combined stroke and death rate was 1.5%, comparable between senior surgeons and trainees (p = 0.60). There was no significant difference in the combined stroke and death rate observed in patients with normal completion angiography (1.4%) compared with that of the patients with a defect corrected (2.8%) (p = 0.28, Odds ratio: 0.67; 95%CI (0.22-2.09)). But there was an increased incidence of postoperative TIA in the group with revision (p = 0.001, odds ratio: 5.8, 95%CI: 1.8-18.9). At 7 years, the freedom rate from >50% carotid restenosis or occlusion was 87.5 ± 6.7% in patients with normal completion angiography and 92 ± 5.4% in patients, who undergo a surgical revision. CONCLUSION: In a single centre, CEA with routine completion angiography resulted in good perioperative outcome. Plaque length, technique for external carotid artery (ECA) endarterectomy and trainee as first operator were independent predictors of operative defects seen on completion angiography.

Protection against Plasmodium falciparum malaria in chimpanzees by immunization with the conserved pre-erythrocytic liver-stage antigen 3.

In humans, sterile immunity against malaria can be consistently induced through exposure to the bites of thousands of irradiated infected mosquitoes. The same level of protection has yet to be achieved using subunit vaccines. Recent studies have indicated an essential function for intrahepatic parasites, the stage after the mosquito bite, and thus for antigens expressed during this stage. We report here the identification of liver-stage antigen 3, which is expressed both in the mosquito and liver-stage parasites. This Plasmodium falciparum 200-kilodalton protein is highly conserved, and showed promising antigenic and immunogenic properties. In chimpanzees (Pan troglodytes), the primates most closely related to humans and that share a similar susceptibility to P. falciparum liver-stage infection, immunization with LSA-3 induced protection against successive heterologous challenges with large numbers of P. falciparum sporozoites.

Factors associated with emergence of pristinamycin-resistant Staphylococcus aureus in a dermatology department: a case-control study.

BACKGROUND: Pristinamycin is used for the treatment of Staphylococcus aureus skin infection. Staphylococcus aureus pristinamycin resistance is usually low. The frequency of pristinamycin-resistant S. aureus (PRSA) increased in the Caen University Hospital dermatology department from 1% in 1998 to >11% in 1999-2002. OBJECTIVES: This study aimed to identify the factors associated with PRSA acquisition. METHODS: Incidences of PRSA and pristinamycin consumption were calculated for the dermatology department and for the rest of the hospital from 1997 to 2007. Individual factors of PRSA acquisition in the dermatology department from 2000 to 2001 were analysed in a retrospective case-control study including 23 cases of PRSA skin colonization or infection and 46 controls with pristinamycin-susceptible S. aureus. Clonal relatedness of isolates was analysed by pulsed-field gel electrophoresis and pristinamycin resistance genes were detected by polymerase chain reaction. Conditional logistic regression was performed to analyse the relationship between pristinamycin resistance and epidemiological and microbiological data. RESULTS: PRSA frequency and pristinamycin consumption were significantly higher in the dermatology department than in other hospital departments. Two epidemic clones of two and six isolates were found for periods of 1 and 2 months, respectively. Thirteen of the 23 PRSA isolates (57%), including all isolates of the two epidemic clones, were found 48 h after the hospitalization or later. PRSA was associated with pristinamycin use during the previous year [odds ratio (OR) 5.60, 95% confidence interval (CI) 1.41-22.22], cumulative use of antibiotics exceeding 1 week during the previous year (OR 4.63, 95% CI 1.47-14.54) and methicillin resistance (OR 6.35, 95% CI 1.38-29.15). CONCLUSIONS: Results suggest that antimicrobial selective pressure and microbial cross-transmission are involved in PRSA acquisition.

A novel stent for percutaneous heart valve implantation: first in vitro results.

Percutaneous aortic valve implantation has become an alternative technique to surgical valve replacement in patients with high risk for surgery. This technique is at its beginning and stents used for valve prostheses remain standard vascular stents. These stents are, however, not designed to undergo heart valve stress. They do not match the aortic environment geometry, and induce exaggerated tissue traumatism. Reduced implant lifetime may therefore be expected. The purpose of the present work is to evaluate in vitro the technical feasibility of noninvasive aortic valve replacement with a novel more specific stent. This stent is especially adapted to its implantation environment with a design that matches the shape of the aortic root while respecting the valve functions. We present a design, a manufacturing process and in vitro performances for the stent under static pressure loading and pulsatile flow. The stent shows good dynamic behavior in keeping position imposed at implantation time and in matching the aortic root dimensions changes. Prosthesis static and dynamic regurgitation are evaluated and show values close to those obtained with other commercially available prostheses.

Solidification mechanisms of chitosan-glycerol phosphate/blood implant for articular cartilage repair.

OBJECTIVE: Chitosan-glycerol phosphate (chitosan-GP) is a unique polymer solution that is mixed with whole blood and solidified over microfractured or drilled articular cartilage defects in order to elicit a more hyaline repair cartilage. For clinical ease-of-use, a faster in situ solidification is preferred. Therefore, we investigated the mechanisms underlying chitosan-GP/blood implant solidification. METHODS: In vitro solidification of chitosan-GP/blood mixtures, with or without added clotting factors, was evaluated by thromboelastography. Serum was analyzed for the onset of thrombin, platelet, and FXIII activation. In vivo solidification of chitosan-GP/blood mixtures, with and without clotting factors, was evaluated in microdrilled cartilage defects of adult rabbits (N=41 defects). RESULTS: Chitosan-GP/blood clots solidified in an atypical biphasic manner, with higher initial viscosity and minor platelet activation followed by the development of clot tensile strength concomitant with thrombin generation, burst platelet and FXIII activation. Whole blood and chitosan-GP/blood clots developed a similar final clot tensile strength, while polymer-blood clots showed a unique, sustained platelet factor release and greater resistance to lysis by tissue plasminogen activator. Thrombin, tissue factor (TF), and recombinant human activated factor VII (rhFVIIa) accelerated chitosan-GP/blood solidification in vitro (P<0.05). Pre-application of thrombin or rhFVIIa+TF to the surface of drilled cartilage defects accelerated implant solidification in vivo (P<0.05). CONCLUSIONS: Chitosan-GP/blood implants solidify through coagulation mechanisms involving thrombin generation, platelet activation and fibrin polymerization, leading to a dual fibrin-polysaccharide clot scaffold that resists lysis and is physically more stable than normal blood clots. Clotting factors have the potential to enhance the practical use, the residency, and therapeutic activity of polymer-blood implants.

Uptake and anterograde axonal transport of wheat germ agglutinin from retina to optic tectum in the chick.

The uptake and anterograde axonal transport of 125I-wheat germ agglutinin (WGA) has been investigated in the visual system of the chick. In order to obtain a marker with specific and homogeneous binding properties, the iodinated lectin was affinity purified by passage over an N-acetylglucosamine (NAcGlu)-Sepharose column after iodination. 22 h after vitreal injection of the purified 125I-WGA, radioactive label was found accumulated in the retinoreceptive layers of the contralateral optic tectum. Gel electrophoresis of tectal homogenates revealed that greater than 80% of the retrieved label ran in a band which comigrated with native WGA. In chicks injected with the fraction of the iodinated preparation that failed to bind to the affinity column, there was no evidence of tectal labeling. These findings support the hypothesis that WGA is selectively taken up by chick retinal ganglion cells and transported intact in an anterograde direction to their axon terminals in the contralateral optic tectum. This raises the possibility that constituents of perikaryal membrane, i.e., lectin receptors, are transported in an anterograde direction by chick retinal ganglion cells.

Prosthetic carotid bypass grafts for atherosclerotic lesions: a prospective study of 198 consecutive cases.

OBJECTIVES: Carotid endarterectomy (CEA) is the standard treatment for atherosclerotic lesions involving the carotid bifurcation. However, CEA can be challenging under some conditions. The goal of this study was to determine the outcome and durability of prosthetic carotid bypass grafting (PCB) with polytetrafluoroethylene (PTFE) grafts as an alternative to CEA. METHODS: This is a prospective series of 198 consecutive patients with PCB, representing 12.4% of 1595 patients with a carotid reconstruction procedure performed in our department between September 1986 and December 2006. Qualifying event was stroke in 67 patients (34%) and transient ischaemic attack (TIA) in 45 (23%), and 86 patients (43%) were asymptomatic. Primary indications for PCB were extensive atherosclerotic lesions (n=71; 36%), carotid stenosis associated with kinking (n=49; 25%), recurrent stenosis (n=47; 23%), stenosis after radiation therapy (n=18; 9%) and technical failure of CEA (n=13; 7%), with excessive arterial wall thinning and perforation after endarterectomy (n=10) or intimal flap on completion digital angiography (n=3). RESULTS: The combined stroke and death rate at 30 days were 0.5% (one stroke). Median follow-up was 9.5 years (interquartile range (IQR): 6.2-18.3 years). At 10 years, primary patency was 97.9+/-3.4%. Six PCBs (3.0%) became occluded during follow-up; one patient had a restenosis greater than 50% and 18 patients (9.1%) had a restenosis of less than 50%. Five patients had an ipsilateral stroke (one postoperative stroke, one at 103 days with a patent PCB and three related to occlusion of the PCB at 4, 13 and 15 years after the procedure). At 10 years, cumulative stroke-free survival was 98.4+/-3.2%, and cumulative survival was 78.8+/-7.0%. CONCLUSIONS: PCB is a safe surgical alternative and is durable, with a low incidence of graft restenosis, when CEA seems hazardous.

Maintaining the knowledge and neonatal resuscitation skills of student midwives 6 months after an educational program.

BACKGROUND: Training student midwives in neonatal resuscitation is essential because the midwife is present at every birth and must be able to perform resuscitation procedures when needed. The objective of this study was to evaluate student midwives' retention of theoretical knowledge about resuscitation as well as their practical application of that knowledge 6 months after training. METHOD: A cross-sectional study was conducted for two consecutive years, 2015 and 2016, among 49 student midwives in the middle of their second and final year of training at the University of Applied Sciences in Western Switzerland. The study included assessments of both theoretical knowledge and practical skills regarding neonatal resuscitation 6 months after each participant had completed the training program. The students' theoretical knowledge was evaluated using a multiple-choice question (MCQ) test, the results of which were compared with results from the same MCQ test that had been obtained 6 months earlier. The students' practical skills were evaluated following a simulation workshop by analyzing recorded videos and applying scores using an adapted validated grid. RESULTS: The MCQ pretest and MCQ posttest (after 6 months) scores showed no statistically significant difference (z=-1.583, P=0.113). In terms of the practical skills assessment, 25% of the students (11/44) were considered insufficiently skilled for the table preparation. During the simulation, 22 teams of students were available for analysis. Of these, 11 teams (50.0%) were considered insufficiently skilled. The skills observation findings showed an integration of technical gestures for the majority of the teams, but the items demonstrated a lack of organization and management. CONCLUSION: Neonatal resuscitation training for student midwives shows that their theoretical knowledge seems to be well assimilated while practical skills are unevenly retained. Certain elements, as indicated by the detail of the analysis grid, need to be reinforced by additional workshops before the end of the training.

Evaluation of an in vitro mast cell degranulation test in the context of food allergy to wheat.

BACKGROUND: Antigenic profiles obtained by ELISA with IgE from patients with wheat food allergy (WFA) established that major allergens are albumins/globulins (AG) for children suffering from atopic eczema/dermatitis syndrome (AEDS), omega5-gliadins for adults suffering from wheat-dependent exercise-induced anaphylaxis (WDEIA), anaphylaxis or urticaria and low-molecular-weight (LMW) glutenin subunits for patients with anaphylaxis. We aimed to characterize a new mast cell transfectant for its ability to degranulate with wheat proteins and patient sera and compare these results to those obtained by ELISA. METHODS: Thirty sera from patients with WFA were tested: 14 with AEDS (group 1) and 16 with WDEIA, anaphylaxis or urticaria (group 2). An IgE Fc receptor (FcepsilonRI) humanized rat RBL-2H3 line was established by transfection with cDNAs encoding alpha-, beta- and gamma-subunits for the human IgE receptor. RESULTS: A humanized RBL clone was selected for its capacity to express mRNA alpha-, beta- and gamma-subunits of FcepsilonRI, to bind allergen-specific human IgE and to degranulate. In group 1, sera induced enhanced degranulation with AG extract, but rarely reacted with gliadins and glutenins. In group 2, half of the sera showed degranulation with LMW glutenins whereas the AG fraction and lipid transfer proteins were rarely positive. omega5-Gliadins did not appear as a major allergen in degranulation assays, although functional allergen-specific IgE was measurable in appreciable amounts. CONCLUSION: Our data demonstrate that in wheat food allergen evaluation, correlation exists between mast cell degranulation and IgE measurements, depending on the type of allergen. Therefore, the biological activity of some allergen types may also be affected by other parameters.

Assessing toxicity of metal contaminated soil from glassworks sites with a battery of biotests.

The present study addresses toxicological properties of metal contaminated soils, using glassworks sites in south-eastern Sweden as study objects. Soil from five selected glassworks sites as well as from nearby reference areas were analysed for total and water-soluble metal concentrations and general geochemical parameters. A battery of biotests was then applied to assess the toxicity of the glassworks soil environments: a test of phytotoxicity with garden cress (Lepidium sativum); the BioTox™ test for toxicity to bacteria using Vibrio fischeri; and analyses of abundancies and biomass of nematodes and enchytraeids. The glassworks- and reference areas were comparable with respect to pH and the content of organic matter and nutrients (C, N, P), but total metal concentrations (Pb, As, Ba, Cd and Zn) were significantly higher at the former sites. Higher metal concentrations in the water-soluble fraction were also observed, even though these concentrations were low compared to the total ones. Nevertheless, toxicity of the glassworks soils was not detected by the two ex situ tests; inhibition of light emission by V. fischeri could not be seen, nor was an effect seen on the growth of L. sativum. A decrease in enchytraeid and nematode abundance and biomass was, however, observed for the landfill soils as compared to reference soils, implying in situ toxicity to soil-inhabiting organisms. The confirmation of in situ bioavailability and negative effects motivates additional studies of the risk posed to humans of the glassworks villages.

Oxidative stress implication in a new phenotype of amyotrophic quadricipital syndrome with cardiac involvement due to lamin A/C mutation.

This study aimed at evaluating OS in an amyotrophic quadricipital syndrome with cardiac impairment in a family of 80 members with a mutation in lamin A/C gene. Twelve patients had cardiac involvement (5 cardiac and skeletal muscles impairment). OS was evaluated in blood samples (thiobarbituric acid-reactive substances (TBARS), carbonylated proteins (PCO)) 6 "affected patients" with phenotypic and genotypic abnormalities without heart failure and 3 "healthy carrier" patients. OS was higher in affected patients than in healthy, as shown by the higher TBARS and PCO values. Patients with cardiac and peripheral myopathy exhibited a higher OS than patients with only cardiac disease (TBARS: 1.73 +/- 0.05 vs. 1.51 +/- 0.04 mmol/l (p = 0.051), PCO: 2.73 +/- 0.34 vs. 0.90 +/- 0.10 nmol/mg protein (p = 0.47)), and with healthy carriers patients (TBARS: 1.73 +/- 0.05 vs. 1.16 +/- 0.14 mmol/l (p = 0.05), PCO: 2.73 +/- 0.34 vs. 0.90 +/- 0.20 nmol/mg protein (p = 0.47)). OS may thus contribute to the degenerative process of this laminopathy. ROS production occurs, prior to heart failure symptoms. We suggest that the extent activation may also promote the variable phenotypic expression of the disease.