[Biomarkers of alcohol consumption, an update]. / Biomarkers voor alcoholgebruik.

Alcohol disorder is a growing public health concern, both national and globally. Accurate assessment using standardized questionnaires is an important step towards effective management of the patient suffering from alcohol disorder. The use of biochemical laboratory measures often helps in obtaining independent estimations of alcohol use. Because of (very) low specificity and sensitivity, the use of traditional biomarkers ASAT, ALAT, ASAT-ALAT ratio and MCV is not recommended anymore in the Netherlands. Use of BAC is recommended to detect recent drinking as is CDT measurement in blood, possibly together with gGT, for the diagnosis of chronic excessive alcohol use. EtG in urine can be used for short term monitoring a patient in an abstinence traject. PEth in blood is a very promising biomarker in detecting recent and chronic alcohol consumption as well as monitoring abstinence. The availability of a fast and cheap method for detecting PEth with a well-defined and accepted cut off, are essential study objectives.

Checklist of the vascular flora of the Sunda-Sahul Convergence Zone.

Background The Sunda-Sahul Convergence Zone, defined here as the area comprising Australia, New Guinea, and Southeast Asia (Indonesia to Myanmar), straddles the Sunda and Sahul continental shelves and is one of the most biogeographically famous and important regions in the world. Floristically, it is thought to harbour a large amount of the world's diversity. Despite the importance of the area, a checklist of the flora has never before been published. Here we present the first working checklist of vascular plants for the Sunda-Sahul Convergence Zone. The list was compiled from 24 flora volumes, online databases and unpublished plot data. Taxonomic nomenclature was updated, and each species was coded into nested biogeographic regions. The list includes 60,415 species in 5,135 genera and 363 families of vascular plants. New information This is the first species-level checklist of the region and presents an updated census of the region's floristic biodiversity. The checklist confirms that species richness of the SSCZ is comparable to that of the Neotropics, and highlights areas in need of further documentation and taxonomic work. This checklist provides a novel dataset for studying floristic ecology and evolution in this biogeographically important region of very high global biodiversity.

Pharmacokinetic/pharmacodynamic analysis of adjunctive perampanel in subjects with partial-onset seizures.

OBJECTIVES: Explore perampanel pharmacokinetics (PK) in all subjects (aged ≥12 years) vs adolescents (aged ≥12 to ≤17 years) with partial-onset seizures (POS) and identify factors explaining between-subject variability in efficacy using a population PK/pharmacodynamic (PD) analysis. MATERIALS & METHODS: Population PK analysis was performed using nonlinear mixed-effect modeling with data from phase II/III randomized, double-blind, placebo-controlled studies of adjunctive perampanel in POS. Perampanel exposure was predicted for all subjects and adolescents. Population PK/PD analyses were performed using data from phase III studies to explore the relationship between perampanel exposure and 28-day average seizure frequency and responder probability. RESULTS: Pooled perampanel PK data from 1318 subjects were described by a one-compartment disposition model. In the absence of antiepileptic drugs (AEDs) affecting perampanel PK, estimated perampanel apparent clearance (CL/F) was 0.668 L/h (all subjects) and 0.682 L/h (adolescent subjects). Co-administration of carbamazepine and oxcarbazepine/phenytoin reduced perampanel exposure. Gender, Asian race (excluding Japanese or Chinese), and increasing alanine aminotransferase lowered perampanel CL/F, but differences were small and not considered clinically relevant. Adolescent outcomes were similar to the total population. Based on PK/PD data from 1748 subjects, percent reduction in 28-day average seizure frequency from baseline and responder probability increased with increasing perampanel exposure; concomitant CYP3A-inducing AEDs lowered perampanel exposure but did not impact the slope for responder probability. CONCLUSIONS: These results are consistent with previous analyses but expand on these through inclusion of a larger number of patients from different ethnic groups, and demonstrate that outcomes were similar between adults and adolescents.

NAD9/NAD7 (mitochondrial nicotinamide adenine dinucleotide dehydrogenase gene)-A new "Holy Grail" phylogenetic and DNA-barcoding marker for Arcellinida (Amoebozoa)?

Molecular phylogeny is an indispensable tool for assessing evolutionary relationships among protists. The most commonly used marker is the small subunit ribosomal RNA gene, a conserved gene present in many copies in the nuclear genomes. However, this marker is not variable enough at a fine-level taxonomic scale, and intra-genomic polymorphism has already been reported. Finding a marker that could be useful at both deep and fine taxonomic resolution levels seemed like a utopic dream. We designed Amoebozoa-specific primers to amplify a region including partial sequences of two subunits of the mitochondrial nicotinamide adenine dinucleotide dehydrogenase gene (NAD9/NAD7). We applied them to arcellinids belonging to distantly related genera (Arcella, Difflugia, Netzelia and Hyalosphenia) and to Arcellinid-rich environmental samples to obtain additional Amoebozoa sequences. Tree topology was congruent with previous phylogenies, all nodes being highly supported, suggesting that this marker is well-suited for deep phylogenies in Arcellinida and perhaps Amoebozoa. Furthermore, it enabled discrimination of close-related taxa. This short genetic marker (ca. 250bp) can therefore be used at different taxonomic levels, due to a fast-varying intergenic region presenting either a small intergenic sequence or an overlap, depending on the species.

[Melanoma of the anal margin]. / Mélanome de la marge anale.

BACKGROUND: Primary anal mucosal melanoma is rare and is associated with a poor prognosis. The observation of a case of anal melanoma at a localized stage in a woman led us to analyze recent data from the literature on therapeutic alternatives. PATIENTS AND METHODS: A 49-year-old woman presented with a pigmented swelling of the anal margin that had begun three months earlier. Complete local excision of the tumour was performed with the conservation of the anal sphincters. Histological examination revealed SSM mucosal melanoma. Abdominoperineal resection was finally performed because of tumoural invasion of the lateral margins. Staging assessment was normal. Half-yearly MRI monitoring of the pelvis was proposed and at nine months no relapse was seen. DISCUSSION: The unusual and misleading symptoms often account for the late diagnosis and poor prognosis of anal melanoma. Treatment is not well defined: local excision with conservation of the anal sphincters is recommended as first-line therapy, but the surgical technique is controversial. Abdominoperineal resection is recommended if the surgical margins are invaded, in the case of local recurrence or if the tumour is inaccessible. The place of adjuvant therapies remains to be defined. More recently, the discovery of mutation in c-KIT mucosal melanoma has allowed the use of biotherapy. Our observation underscores the importance of early detection of anal melanoma by all practitioners concerned in view of its aggressiveness and we report the difficulties of therapeutic management in the absence of established guidelines.

Mumps: burden of disease in France.

This article provides a review of the epidemiological data on mumps in France since 1986. The results of 26 years of monitoring in general practice by the Sentinel network are analysed, such as hospitalisation data between 2004 and 2010, as well as mortality data between 2000 and 2009. The annual incidence rate has plummeted between 1986 and 2011, from 859 cases per 100,000 inhabitants [95% CI: 798-920] to 9 cases per 100,000 inhabitants [95% CI: 4-14]. A change in the age distribution is significant with an increase of Relative Illness Ratio (RIR) for patients over 20 years. Since 2000, vaccine status has also changed, and the majority of recent mumps cases occur among previously vaccinated patients. The average annual hospitalisation rate is 3.2 per 1 million inhabitants. Mumps was identified as the initial cause of death in 1 case every 5 years. This study estimates the burden of mumps disease in France.

Diagnostic value of zinc protoporphyrin in a screening strategy for alpha-thalassemia.

The definitive diagnosis of alpha-thalassemia involves detection of a deletion of one or more alpha-globin that encode the alpha-chains of Hb (hemoglobin). To determine whether DNA analysis is indicated, screening tests such as mean corpuscular volume (MCV) and Hb typing are employed. alpha-Thalassemia often correlates with normal or low HbA2 values. Zinc protoporphyrin (ZPP) is usually high in ferropenic anemia or lead-poisoning and is normal or slightly raised in beta-thalassemia. Therefore, ZPP is currently used as a marker to discriminate between ferropenic anemia and beta-thalassemia. We investigated the diagnostic potential of ZPP < 150 micromol/mol heme in a screening strategy for alpha-thalassemia. We measured ZPP and performed DNA analysis for detecting the seven most prevalent alpha-thalassemia deletions, namely, alpha3.7, SEA, alpha20.5, alpha4.2, MED, FIL, and THAI, in the blood samples of 200 patients with MCV < 70 fL and HbA2 < or = 3.5%. Deletions were detected in 9% subjects in the ZPP > or = 150 group (n = 175) and 56% subjects in the ZPP < 150 group (n = 29); this difference was statistically significant (chi-square test, P < 0.001). We conclude that ZPP < 150 micromol/mol heme can be used in a new screening strategy for alpha-thalassemia.

Laboratory evaluation of a novel capillary blood sampling device for measuring eight clinical chemistry parameters and HbA1c.

BACKGROUND: The Demecal set enables medically unskilled persons to produce diluted plasma from a single drop of capillary blood at any time and place. The sample is mailed to a certified laboratory for analysis. A marker compound in the dilution buffer enables to correct for individual blood sampling variation. A test dependent factor corrects for recovery of analytes. METHODS: Glucose, cholesterol, triglycerides, HDL and LDL cholesterol, creatinine, BUN, uric acid and HbA1c were evaluated. We studied the correction procedure for sampling variation by the marker compound, the precision of the analyzer in the low range, the influence of characteristics of the set on analyte recovery, and the stability of the samples at different temperatures. RESULTS: Using the marker compound in the buffer, variation in sampling could be corrected accurately. For dilutions up to 15 times, the precision of the analyzer was sufficient. Application of test specific recovery factors gave a good correlation with results of venous blood samples. Samples were stable for 4 days at 4 degrees C, 2-3 days at room temperature and 1 day at 37 degrees C. CONCLUSIONS: The Demecal set can be considered an alternative for venous blood sampling for the tested parameters, enabling patient friendly management of chronic disease.

A20 deletion is associated with copy number gain at the TNFA/B/C locus and occurs preferentially in translocation-negative MALT lymphoma of the ocular adnexa and salivary glands.

The genetic basis of MALT lymphoma is largely unknown. Characteristic chromosomal translocations are frequently associated with gastric and pulmonary cases, but are rare at other sites. We compared the genetic profiles of 33 ocular adnexal and 25 pulmonary MALT lymphomas by 1 Mb array-comparative genomic hybridization (CGH) and revealed recurrent 6q23 losses and 6p21.2-6p22.1 gains exclusive to ocular cases. High-resolution chromosome 6 tile-path array-CGH identified NF-kappaB inhibitor A20 as the target of 6q23.3 deletion and TNFA/B/C locus as a putative target of 6p21.2-22.1 gain. Interphase fluorescence in situ hybridization showed that A20 deletion occurred in MALT lymphoma of the ocular adnexa (8/42=19%), salivary gland (2/24=8%), thyroid (1/9=11%) and liver (1/2), but not in the lung (26), stomach (45) and skin (13). Homozygous deletion was observed in three cases. A20 deletion and TNFA/B/C gain were significantly associated (p<0.001) and exclusively found in cases without characteristic translocation. In ocular cases, A20 deletion was associated with concurrent involvement of different adnexal tissues or extraocular sites at diagnosis (p=0.007), a higher proportion of relapse (67% versus 37%) and a shorter relapse-free survival (p=0.033). A20 deletion and gain at TNFA/B/C locus may thus play an important role in the development of translocation-negative MALT lymphoma.

The beta and gamma classes of carbonic anhydrase.

There are currently five (alpha, beta, gamma, delta, zeta) classes of carbonic anhydrases (CA's) of which the alpha-class from mammalian sources has been studied to a much greater extent compared to the other four classes. Yet, CA's other than the alpha-class are widely distributed in Nature and play important roles in human health, the global carbon cycle, and industrial applications. In aerobic prokaryotes, beta-class CA's are implicated in maintaining internal pH and CO(2)/bicarbonate balances required for biosynthetic reactions. In anaerobic prokaryotes, beta-class CA's are implicated in the transport of CO(2) and bicarbonate across the cytoplasmic membrane that regulates pH and facilitates acquisition of substrates and product removal required for growth. In phototrophic organisms, beta-class CA's are particularly important for transport and concentration of CO(2) and bicarbonate for photosynthesis. The delta- and zeta-classes are proposed to function in marine diatoms to concentrate CO(2) for photosynthesis. Physiological roles for the gamma-class are not as well documented; however, the active site architecture and catalytic mechanism is well understood as are patterns of inhibition by sulfonamides and anions.

Conjunctival marginal zone B-cell lymphoma (MALT lymphoma) with amyloid and relapse in the stomach.

The authors report a localized (primary) conjunctival marginal zone B-cell lymphoma (mucosa-associated lymphoid tissue (MALT)-type), with amyloid deposition with relapse in the stomach, 14 months after the initial diagnosis. Ocular adnexal marginal zone B-cell MALT lymphoma is often localized at diagnosis; some relapse in typical MALT sites. There are few reports of localized conjunctival lymphoma with a relapse in the stomach. The authors suggest that all patients with localized ocular adnexal lymphoma be followed for an extended period.

Effect of cationic carriers on the pharmacokinetics and tumor localization of nucleic acids after intravenous administration.

Nucleic acid based therapeutics are currently being studied for their application in cancer therapy. In this study, the effect of different cationic delivery systems on the circulation kinetics, tumor localization, and tissue distribution of short interfering RNA (siRNA) and plasmid DNA (pDNA) was examined, after intravenous administration in mice bearing a s.c. Neuro 2A tumor. Nanosized particles were formed upon complexation of siRNA with the cationic liposome formulation DOTAP/DOPE and the targeted, cationic polymer RGD-PEG-PEI. Both the circulation kinetics and the overall tumor localization of the siRNA complexes were similar to non-complexed siRNA. Importantly, the different carriers changed the intratumoral distribution of siRNA within the tumor. pDNA was effectively condensed with linear polyethylenimine (PEI), PEGylated linear PEI (PEG-PEI) or poly(2-dimethylamino ethylamino)phosphazene. Only PEG-PEI was able to improve the pDNA circulation kinetics. All pDNA complexes yielded similar pDNA tumor localization (1% of the injected dose, 60 min after administration). We conclude that the level of nucleic acid tumor localization is independent on the type of formulation used in this study. Therefore, the value of carrier systems for the intravenous delivery of nucleic acids cannot be solely attributed to benefits relevant during the transport towards the tumor. Rather, the benefits are arising from carrier-induced changes in the intratumoral fate of the nucleic acids.

Suture anchors are superior to transglenoid sutures in arthroscopic shoulder stabilization.

PURPOSE: We retrospectively compared 2 groups of high-demand patients with post-traumatic anterior shoulder instability to determine whether arthroscopic stabilization was superior with transglenoid suture or suture anchors. METHODS: In a retrospective comparative study we investigated the results of 246 high-demand patients, with post-traumatic anterior shoulder instability, who underwent arthroscopic capsulolabral reconstruction: 165 (mean age, 27.5 years; mean follow-up, 80 months) were evaluated after treatment with transglenoid sutures, and 81 (mean age, 26.6 years; mean follow-up, 27 months) were treated with suture anchors in a consecutive period. We compared both techniques with regard to recurrence rate, postoperative complications, range of motion, sport activity, work, and patient satisfaction. RESULTS: In the anchor group recurrent dislocation after surgery occurred in 7 patients (8.7%), all within 18 months postoperatively. This finding was significantly (P = .009) better than that in the transglenoid group, in which recurrent postoperative dislocation occurred in 57 patients (34%), in a period of 0 to 115 months after surgery. Postoperative complications were seen in 4 of 81 patients in the suture anchor group, whereas a significantly (P = .01) higher rate was found in the transglenoid suture group, with 36 complications in 35 of 165 patients. CONCLUSIONS: The data presented in this study suggest that the modern suture anchor technique results in a better outcome after shoulder stabilization, with fewer complications and lower recurrence rates, than the transglenoid repair. We conclude that the suture anchor technique should be a preferred method for arthroscopic shoulder stabilization surgery. LEVEL OF EVIDENCE: Level III, retrospective, comparative therapeutic study.

[Main issues and contributions of the evaluation of health care networks]. / Les problématiques et les apports de l'evaluation d'un réseau de santé.

Health care networks correspond to a form of horizontal and flexible organisation, which consists in dealing with complex problems of health and long-term care. Networks rest entirely on the mechanisms of coordination between the various actors of care, whose keystone is the medical information flow. The challenge of the health networks evaluation is to prove the added value of a network organization, instead of a conventional disease-centred health care. The evaluation of health care networks is legally required but not sustained by specialists. Evaluation must ensure the existence of a real management; measure the interest of health care professionals and the impact of the network on professional practices. Moreover, evaluation accompanies the development of the network and offers the advisability to the decision maker of obtaining a tool for control of management, i.e. an information tool and a decision-making help, with the aim of a continuous improvement of the quality of care. However the absence of formation of the medical coordinators to management, makes networks management problematic. Thus, health care networks cannot make the saving in a good administrative and management base. In the same way health care networks must obtain an information system adapted to their type of coordination, allowing the evaluation required by the financier.

Application of poly(2-(dimethylamino)ethyl methacrylate)-based polyplexes for gene transfer into human ovarian carcinoma cells.

Previously, attempts were made in our laboratory to transfect human ovarian cancer (OVCAR-3) cells, growing in the peritoneal cavity of nude mice, by intraperitoneal administration of poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA)-based polyplexes. However, hardly any transfection of the OVCAR-3 cells was observed. The aim of the present study was to examine whether pDMAEMA-polyplexes can transfect OVCAR-3 cells in vivo at DNA doses much higher than used previously [J. Gene Med. 1 (1999) 156-158]. We also explored a specific targeting strategy based on the use of folic acid (FA) as a targeting ligand directed against the folate receptor overexpressed on OVCAR-3 cells. Luciferase expression by OVCAR-3 cells mediated by pDMAEMA-based polyplexes was evaluated in the mouse i.p. OVCAR-3 xenograft model of ovarian cancer. By virtue of new formulation options, we were able to administer polyplex dispersions into OVCAR-3 bearing mice at much larger doses (75-120 microg DNA) than used previously (15 microg). The feasibility of folate-mediated targeting of the polyplexes was studied after coupling of FA to preformed polyplexes with poly(ethylene glycol) (PEG) as a spacer. Intraperitoneal administration of naked pLuc plasmid did not result in significant gene expression by the tumor cells. Administration of uncoated, positively charged pDMAEMA-based polyplexes at a DNA dose of 75-120 microg yielded significant transfection activity. However, also considerable gene expression was observed in non-target cells. To avoid transfection of non-target cells, an active targeting strategy based on the use of FA was studied. At a dose of 75 microg DNA (N/P 5), the folate-targeting approach yielded about 10-fold lower luciferase transfection levels in organs lined by the mesenthelial layer. This beneficial site-avoidance effect was achieved without compromising the degree of tumor cell transfection. Successful transfection of OVCAR-3 cells growing in the peritoneal cavity of nude mice can be achieved by i.p. administration of polyplexes at doses between 75 and 120 microg DNA. It was further demonstrated that active targeting of polyplexes to OVCAR-3 cells growing in the peritoneal cavity of mice is a realistic possibility to avoid transfection of non-target cells.

Epiphytism and pollinator specialization: drivers for orchid diversity?

Epiphytes are a characteristic component of tropical rainforests. Out of the 25,000 orchid species currently known to science, more than 70% live in tree canopies. Understanding when and how these orchids diversified is vital to understanding the history of epiphytic biomes. We investigated whether orchids managed to radiate so explosively owing to their predominantly epiphytic habit and/or their specialized pollinator systems by testing these hypotheses from a statistical and phylogenetic standpoint. For the first approach, species numbers of 100 randomly chosen epiphytic and terrestrial genera were compared. Furthermore, the mean number of pollinators per orchid species within the five subfamilies was calculated and correlated with their time of diversification and species richness. In the second approach, molecular epiphytic orchid phylogenies were screened for clades with specific suites of epiphytic adaptations. Epiphytic genera were found to be significantly richer in species than terrestrial genera both for orchids and non-orchids. No evidence was found for a positive association between pollinator specialization and orchid species richness. Repeated associations between a small body size, short life cycle and specialized clinging roots of twig epiphytes in Bulbophyllinae and Oncidiinae were discovered. The development of twig epiphytism in the first group seems repeatedly correlated with speciation bursts.

Merits of polyoxotungstates as environmental remediation catalysts: a novel wet oxidation technology for refractory industrial pollutants.

The catalytic effect of silicadodecatungstate (SiW12O40(4-)) on the oxidation of nitrobenzene in subcritical water (T = 100-300 degrees C; P = 0.1-8.6MPa, pH = 2.6) was examined. In the absence of SiW12O40(4-) catalyst nitrobenzene oxidation slowed down significantly after a brief period of rapid degradation and an average of 40% nitrobenzene removal. The catalytic effect of SiW12O40(4-) becomes particularly pronounced after a temperature dependent induction period. Catalyzed wet air oxidation continued until the degradation of the parent compound was complete provided that experimental conditions were harsh enough (T > 200 degrees C). Product distribution analysis suggested that the oxidation paths for catalyzed and uncatalyzed wet air oxidation of nitrobenzene are similar, but the analysis was not definitive.