Multidisciplinary care management has a positive effect on paediatric obesity and social and individual factors are associated with better outcomes.

AIM: The RePPOP Aquitaine network, which was established in south-west France to prevent and treat paediatric obesity, has developed a multidisciplinary approach based on multicomponent lifestyle interventions and family-based actions. This study assessed the impact of its care management programme and investigated the factors associated with better outcomes. METHODS: The impact of the RePPOP care management programme was assessed by changes in the body mass index (BMI) Z score, between baseline and the end-of-care management. We focused on 982 overweight or obese children (59.9% girls) with a mean age of 10.64 years and a range of 2.4-17.9 years. A multivariate analysis examined the independent factors associated with better outcomes. RESULTS: At the end-of-care management programme, 75.5% of children had decreased their BMI Z score. Initial characteristics significantly associated with better outcomes were as follows: the age at baseline between five years and 15 years of age, playing sport at a club, being followed up by RePPOP for longer than 10 months, no parental obesity and no academic difficulties. CONCLUSION: This study confirmed that multidisciplinary treatment had a significant positive effect on paediatric obesity and that social and individual factors affected the efficiency of the care management.

Overnight improvements in two REM sleep-sensitive tasks are associated with both REM and NREM sleep changes, sleep spindle features, and awakenings for dream recall.

Memory consolidation is associated with sleep physiology but the contribution of specific sleep stages remains controversial. To clarify the contribution of REM sleep, participants were administered two REM sleep-sensitive tasks to determine if associated changes occurred only in REM sleep. Twenty-two participants (7 men) were administered the Corsi Block Tapping and Tower of Hanoi tasks prior to and again after a night of sleep. Task improvers and non-improvers were compared for sleep structure, sleep spindles, and dream recall. Control participants (N = 15) completed the tasks twice during the day without intervening sleep. Overnight Corsi Block improvement was associated with more REM sleep whereas Tower of Hanoi improvement was associated with more N2 sleep. Corsi Block improvement correlated positively with %REM sleep and Tower of Hanoi improvement with %N2 sleep. Post-hoc analyses suggest Tower of Hanoi effects-but not Corsi Block effects-are due to trait differences. Sleep spindle density was associated with Tower of Hanoi improvement whereas spindle amplitude correlated with Corsi Block improvement. Number of REM awakenings for dream reporting (but not dream recall per se) was associated with Corsi Block, but not Tower of Hanoi, improvement but was confounded with REM sleep time. This non-replication of one of 2 REM-sensitive task effects challenges both 'dual-process' and 'sequential' or 'sleep organization' models of sleep-dependent learning and points rather to capacity limitations on REM sleep. Experimental awakenings for sampling dream mentation may not perturb sleep-dependent learning effects; they may even enhance them.

Melanoma incidence and Breslow tumour thickness development in the central Alpine region of South Tyrol from 1998 to 2012: a population-based study.

BACKGROUND: Cutaneous melanoma incidence is increasing worldwide, especially for in situ and thin (<1 mm) lesions, while thick (≥1 mm) lesions have been generally stable in many studies; although epidemiological data on melanoma is readily available, population-based studies, especially on mountain regions, are rare. OBJECTIVE: The aim of this study was to analyse cutaneous melanoma incidence and Breslow tumour thickness in the central Alpine mountain region of South Tyrol, northern Italy. METHODS: All newly diagnosed cutaneous in situ and invasive melanomas in the resident population from 1998 to 2012 were taken from the Pathology Unit, Bolzano Hospital and South Tyrol Cancer Registry. Incidence and Breslow tumour thickness were analyzed. Statistical analyses included Mann-Whitney and Kruskal-Wallis tests. RESULTS: A total of 784 in situ melanomas and 1663 invasive melanomas were collected. In situ melanomas showed the highest increase: European age-standardized incidence rose from 2.44 per 100,000 inhabitants in 1998-2002 to 16.01 in 2008-2012. Invasive melanoma incidence increased from 12.69 in 1998-2002, to 22.88 in 2008-2012. The incidence rise was observed in thin melanomas (from 8.39 to 16.18) and in thick melanomas (from 4.30 to 6.70). Breslow distribution revealed a median value of 0.62 mm (mean 1.34; SD 2.24; range 0.09-30.0) and did not change significantly over time (P = 0.286). Higher Breslow values were observed at advanced age (P < 0.001), among males (P = 0.017), in nodular melanomas (P < 0.001) and at higher Clark levels (P < 0.001). Significant differences were also found in urban hospitals compared to rural hospitals during the whole period (P = 0.004), but not in the last 5 years (P = 0.053). CONCLUSION: Incidence of cutaneous melanoma is increasing in South Tyrol, especially for in situ and thin lesions, but also for thick lesions; no reduction in median tumour thickness is observed. Rural areas and elevated altitudes may contribute to this effect.

Differential effects of sleep deprivation on sleepwalking: Role of demographic and clinical profiles.

BACKGROUND: Although sleepwalking is one of the most prevalent and potentially injurious of the NREM parasomnias, it is still diagnosed primarily based on the patient's clinical history. Early pilot work suggested that sleep deprivation protocols could help obtain a polysomnographically-based (PSG) diagnosis of sleepwalking, but larger studies remain lacking. METHODS: We compared baseline PSG recordings with those obtained after 25hrs of sleep deprivation in a cohort of 124 consecutively assessed adult sleepwalkers. RESULTS: When compared to baseline recordings, post-sleep deprivation PSG assessments resulted in nearly twice as many somnambulistic episodes being recorded in the laboratory and significantly increased the proportion of patients (from 48 % to 63 %) experiencing at least one lab-based episode. Moreover, while 17 % of patients experienced a sleepwalking event exclusively during recovery sleep, only 2 % of patients did so solely at baseline. Sleep deprivation had similar facilitating effects on patents' somnambulistic events regardless of age of onset and positive versus negative family history for sleepwalking. Younger age and higher home episode frequency both predicted a positive response to sleep deprivation. A separate group of 17 patients with comorbid sleep disorders showed a similar increase in their proportion experiencing at least one episode during recovery sleep. CONCLUSION: The results from this large series of sleepwalkers provide strong support for the use of sleep deprivation in facilitating the occurrence of somnambulistic events in the sleep laboratory.

[Evolution of food supply (apart from school catering) between 2004/2005 and 2009/2010 in middle- and high-schools of Aquitaine, France]. / Évolution de l'offre alimentaire hors restauration scolaire entre 2004-2005 et 2009-2010 dans les établissements du second degré d'Aquitaine.

BACKGROUND: The aim of this study was to assess the evolution of food supply (apart from school catering) between school years 2004/2005 and 2009/2010, in middle- and high-schools from the Aquitaine region (southwest France), in order to evaluate the impact of actions conducted within the framework and the program Nutrition, Prevention and Health of children and adolescents in Aquitaine (southwest France). METHODS: Two surveys were carried out among all middle- and high-schools of the Aquitaine region in 2004/2005 (n=536) and 2009/2010 (n=539) within the framework of a regional multidisciplinary public health program "Nutrition, prevention and health of children and teenagers in Aquitaine". For both 2004/2005 and 2009/2010, data were collected using the same questionnaire and dealt with school characteristics and modalities of food supply (apart from school catering). RESULTS: Response rate was 84.1% in 2004/2005 and 79.6% in 2009/2010. The proportion of schools offering food to pupils (apart from school catering) significantly decreased in 5 years (from 80.1% to 50.1%, P<0.001). Between 2004/2005 and 2009/2010, we observed a stabilization in the proportion of schools offering free food (from 19.7 to 17%, P=0.3), a significant decrease of those selling food (from 62.8 to 37.1, P<0.001), offering vending machines (from 43.5 to 7.2, P<0.001) and a significant increase of those offering water supply (from 8.2 to 44%, P<0.001). The composition of each modality of food supply (apart from school catering) has also been improved: less sweet and fat food, more bread and fruits. CONCLUSION: This study shows an overall improvement of food supply apart from school catering (food sale, free food and vending machines) in middle- and high-schools from the Aquitaine region (southwest France) between 2004/2005 and 2009/2010. This improvement is related to the proportion of schools offering food (quantitative improvement), as well as to the composition of food supply (qualitative improvement). These results show an improvement of food supply (apart from school catering), suggesting that actions implemented in the framework of the program "Nutrition, prevention and health of children and adolescents in Aquitaine" may have led to these improvements.

RLIM inhibits functional activity of LIM homeodomain transcription factors via recruitment of the histone deacetylase complex.

LIM domains are required for both inhibitory effects on LIM homeodomain transcription factors and synergistic transcriptional activation events. The inhibitory actions of the LIM domain can often be overcome by the LIM co-regulator known as CLIM2, LDB1 and NLI (referred to hereafter as CLIM2; refs 2-4). The association of the CLIM cofactors with LIM domains does not, however, improve the DNA-binding ability of LIM homeodomain proteins, suggesting the action of a LIM-associated inhibitor factor. Here we present evidence that LIM domains are capable of binding a novel RING-H2 zinc-finger protein, Rlim (for RING finger LIM domain-binding protein), which acts as a negative co-regulator via the recruitment of the Sin3A/histone deacetylase corepressor complex. A corepressor function of RLIM is also suggested by in vivo studies of chick wing development. Overexpression of the gene Rnf12, encoding Rlim, results in phenotypes similar to those observed after inhibition of the LIM homeodomain factor LHX2, which is required for the formation of distal structures along the proximodistal axis, or by overexpression of dominant-negative CLIM1. We conclude that Rlim is a novel corepressor that recruits histone deacetylase-containing complexes to the LIM domain.

Allosteric effects of Pit-1 DNA sites on long-term repression in cell type specification.

Reciprocal gene activation and restriction during cell type differentiation from a common lineage is a hallmark of mammalian organogenesis. A key question, then, is whether a critical transcriptional activator of cell type-specific gene targets can also restrict expression of the same genes in other cell types. Here, we show that whereas the pituitary-specific POU domain factor Pit-1 activates growth hormone gene expression in one cell type, the somatotrope, it restricts its expression from a second cell type, the lactotrope. This distinction depends on a two-base pair spacing in accommodation of the bipartite POU domains on a conserved growth hormone promoter site. The allosteric effect on Pit-1, in combination with other DNA binding factors, results in the recruitment of a corepressor complex, including nuclear receptor corepressor N-CoR, which, unexpectedly, is required for active long-term repression of the growth hormone gene in lactotropes.

[Patients' satisfaction and perinatal network: a preliminary report]. / Pré-étude sur la satisfaction des femmes au sein d'un réseau de santé périnatale.

OBJECTIVES: The objective of this work was to determine the principal axes for measuring the satisfaction of women transferred within a perinatal network. MATERIAL AND METHODS: We began with two successive qualitative sociological approaches (18 semi-structured interviews of women and of couples in 2004 and 2006-2007) and one quantitative approach (questionnaire survey of 583 women hospitalized in 15 of the 16 maternity units in the Auvergne network during a two-week period in 2004). RESULTS: The qualitative surveys show that the procedures at arrival at the new establishment and the feeling of consistency that they did or did not induce, the identification of the participants, an understanding of their role, and the objectives of the new examinations were all important dimensions. During hospitalization, explanations of who does what and the consistency of the information provided also played a role in this satisfaction. The quantitative survey (participation rate=89%) showed that 77% of the women were aware of the possibility of an in utero transfer. Nonetheless, most of them had very little information about the network. CONCLUSION: It is possible to identify specific themes associated with women's satisfaction about in utero transfers that are useful for constructing a questionnaire that must, in turn, be validated.

Relationships between emotional disorders, personality dimensions, and binge eating disorder in French obese adolescents.

PURPOSE: Binge eating disorder (BED) is associated with higher psychopathology, including emotional and personality disorders, in the adult population, whether or not they are obese; although few data are available on adolescents, particularly among obese adolescents. OBJECTIVE: To explore the association of both emotional disorders and personality dimensions with BED in obese adolescents. METHODS: The sample consisted of 115 French adolescents enrolled at a clinical unit for the multidisciplinary care of their overweight or obesity. BED was defined using the Binge Eating Scale (BES). Emotional disorders and personality dimensions were assessed using the following tools: Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Junior Temperament and Character Inventory (JTCI); impulsivity was determined by the Barratt Impulsiveness Scale (BIS) and the Toronto Alexithymia Scale (TAS). Associations between emotional disorders (BDI/BAI) and personality dimensions (TAS/BIS/JTCI), considered first separately and then jointly with BED were determined with multivariate analysis. RESULTS: More severe depression (ß=0.27, CI [0.06; 0.48], P=0.011), a higher level of excess weight (ß=1.91, CI [0.22; 3.59], P=0.027), older age (ß=1.28, CI [0.43; 2.14], P=0.003), and greater cooperativeness (ß=0.36, CI [0.07; 0.66], P=0.017) were independently significantly associated with the presence of BED. CONCLUSIONS: This cross-sectional study underlines the co-occurrence of emotional and personality disorders with BED. This points out the importance of a multidisciplinary approach and the relevance of a joint diagnosis of binge eating, emotional disorders, and personality dimensions in obese adolescents, for better prevention and treatment of pediatric obesity.

Characterization of quail Pax-6 (Pax-QNR) proteins expressed in the neuroretina.

After differential screening of a cDNA library constructed from quail neuroretina cells (QNR) infected with the v-myc-containing avian retrovirus MC29, we have isolated a cDNA clone, Pax-QNR, homologous to the murine Pax-6, which is mutated in the autosomal dominant mutation small eye of mice and in the disorder aniridia in humans. Here we report the characterization of the Pax-QNR proteins expressed in the avian neuroretina. From bacterially expressed Pax-QNR peptides, we obtained rabbit antisera directed against different domains of the protein: paired domain (serum 11), domain between the paired domain and homeodomain (serum 12), homeodomain (serum 13), and carboxyl-terminal part (serum 14). Sera 12, 13, and 14 were able to specifically recognize five proteins (48, 46, 43, 33, and 32 kDa) in the neuroretina. In contrast to proteins of 48, 46, and 43 kDa, proteins of 33 and 32 kDa were not recognized by the paired antiserum (serum 11). Paired-less and paired-containing proteins exhibited the same half-life (6 h) and were phosphorylated mostly on serine residues. Immunoprecipitations performed with subcellular fractions of neuroretinas showed that the paired-containing proteins were located in the nucleus, whereas the 33- and 32-kDa proteins were found essentially in the cytoplasmic compartment. However, immunofluorescence experiments performed after transient transfections showed that p46 and p33/32 were also located in vivo into the nucleus. Thus, the Pax-QNR/Pax-6 gene can produce proteins with two DNA-binding domains as well as proteins containing only the DNA-binding homeodomain.

Link between nasal carriage of Staphylococcus aureus and infected diabetic foot ulcers.

AIMS: Nasal carriage of Staphylococcus aureus in diabetic patients may be a risk factor for diabetic foot lesion infections. The aims of this study were to compare the genotypic profiles of S. aureus strains isolated from nares and diabetic foot ulcers (DFUs) using microarray technology. METHODS: Patients were included if they were admitted for diabetic foot infection (DFI) at any of three diabetology departments of Montpellier and Nîmes University Hospitals between 1 September 2010 to 30 June 2012. All S. aureus isolates were analyzed using oligonucleotides arrays; S. aureus resistance and virulence genes were determined and each isolate was affiliated to a clonal complex. RESULTS: The prevalence of S. aureus nasal carriage among the 276 included patients was 39.5% (n=109), while 36.6% (n=101) had S. aureus at both sites (nares and foot wounds) and, of these patients, 65.3% of patients harboured the same strain at both sites. In addition, the spread of the methicillin-resistant S. aureus (MRSA) ST398 clone in DFI and its tropism for bone were also further confirmed. CONCLUSION: These findings appear to provide new arguments in favour of the systematic detection of nasal S. aureus carriage to anticipate the management of DFI.

Step-by-step build-up of covalent poly(ethylene oxide) nanogel films.

Hydrogels based on poly(ethylene glycol) (PEG) are commonly used for studies related to cell fate and tissue engineering. Here we present a new covalent layer-by-layer build-up process leading to PEG coatings of nanometer size called "nanogel films". Compared to macroscopic hydrogels, such nanogels should provide a fine control over the structure and the thickness of the coating. Alternated deposition of bifunctional and tetra functional PEG molecules reacting through thiol/maleimide click chemistry is evaluated by quartz crystal microbalance. We first study parameters influencing the build-up process of such coatings and demonstrate the importance of (i) the nature of the first deposited layer, (ii) the PEG concentrations and (iii) the length of the PEG chains that appears to be the most significant parameter influencing film growth. The build-up process can be extended to a large variety of substrates like SiO2 or polymers by using an appropriate anchoring layer. Covalent functionalization of these nanogel films by proteins or enzymes is suited by modifying the biomolecules with thiol or maleimide groups and immobilizing them during the build-up process. Activity of the embedded enzymes can be maintained. Moreover ligands like biotin can be incorporated into the film and recognition by streptavidin can be modulated by playing with the number of PEG layers covering biotin. Compared to well-known PEG hydrogels, these new coatings are promising as they allow to (i) build thin nanometric coatings, (ii) finely control the amount of deposited PEG and (iii) organize the position of the embedded biomolecules inside the film layers.

RT-PCR analysis of corticotroph-associated genes expression in carcinoid tumours in the ectopic-ACTH syndrome.

OBJECTIVE: ACTH is frequently produced in non-pituitary tumours, leading to the ectopic-ACTH syndrome, but the molecular mechanisms of its expression remain obscure. This study was aimed at understanding the transcription mechanisms of the ACTH-precursor gene in carcinoid tumours of the lung or thymus. DESIGN: Transcripts coding for a series of corticotroph-associated transcription factor genes were detected, together with markers of the corticotroph phenotype. We studied a series of 41 carcinoid tumours including 15 with proven ectopic-ACTH syndrome. METHODS: Specific RT-PCR reactions were designed for each gene including alternatively spliced isoforms. RESULTS: The markers of the corticotroph phenotype were detected in all ACTH-positive tumours. Expression of the Tpit and Pitx1 genes were not restricted to ACTH-positive tumours but were also detected in many ACTH-negative carcinoids. Only a subset of ACTH-negative tumours expressed NAK-1/Nur77, and NeuroD1 expression was detected in approximately 50% of the tumours regardless of their secretory status. The glucocorticoid receptor alpha was detected in every tumour in contrast to its beta isoform detectable in a few tumours only. Chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TF1) and peroxisome proliferator-activated receptor (PPAR) gamma2 were expressed in 50% of the tumours of each group whereas PPARgamma1 was expressed in almost every tumour. CONCLUSIONS: ACTH-positive carcinoids do not share a characteristic expression pattern of the corticotroph-associated transcription factor genes, suggesting that the transcriptional mechanisms of the ACTH-precursor gene differ from those in normal pituitary corticotrophs. Expression of Tpit and Pitx1 genes in most carcinoids suggests that some aspects of the pituitary corticotroph phenotype may belong to general carcinoid differentiation.

Characterization of a paired box- and homeobox-containing quail gene (Pax-QNR) expressed in the neuroretina.

The retina is an integral part of the central nervous system, and consists of two layers, the outer pigmented layer and the inner sensory layer or neuroretina (NR). The NR layer contains several strata of cells (glial and neuronal) derived from proliferating neuroectodermal precursors that differentiate after terminal mitosis. In vitro, NR cells can differentiate not only into neuronal and glial types, but also into pigment and lens cells. Quail (Coturnix coturnix japonica) NR cells (QNR) infected with MC29 transforming retrovirus become pigmented after several passages in vitro. In order to characterize the genes expressed in these pigmented MC29 QNR, a cDNA library was prepared from these cells. After differential screening we have isolated a cDNA clone which identifies an RNA expressed in NR but not in the pigmented layer of the retina. This cDNA encodes a protein related to that of Drosophila, mouse and zebrafish paired box- and homeobox-containing segmentation genes and is called Pax-QNR. The expression of Pax-QNR in the NR is confined to the ganglionic cell layer and to the lower part of the inner nuclear layer containing the amacrine or correlation neurones.

A model for the development of the hypothalamic-pituitary axis: transcribing the hypophysis.

Mammalian organogenesis involves a sequential program to generate cells with specific fates and phenotypes from a common primordium, which is hypothesized to be the consequence of regulated overlapping patterns of expression of specific sets of transcription factors in a precise spatiotemporal manner. The hypothalamic-pituitary axis is critical for survival and homeostasis, controlling growth, reproduction, metabolism and behavior, and constitutes an ideal model in which to define the molecular markers to emergence of specific cell phenotypes from a common primordium. Development of the anterior pituitary gland is controlled by sequential series of gradients of specific signaling molecules that, in turn, appear to coordinate the expression of specific combinations of transcription factor-encoding genes, many of which as tissue-specific or tissue restricted factors that serially dictate cell-type determination and terminal differentiation events that underlie the differentiated cell phenotype.

Pax-QNR/Pax-6, a paired box- and homeobox-containing gene expressed in neurons, is also expressed in pancreatic endocrine cells.

After differential screening of a cDNA library constructed from quail neuroretina cells infected with the v-myc containing avian retrovirus MC29, we have isolated a cDNA clone Pax-QNR, homologous to the murine Pax6 which is mutated in the autosomal dominant mutation small eye (Sey) of the mouse and aniridia in man. Here we report the characterization of Pax-QNR/Pax-6 expression in the chicken, quail, and mouse pancreas. In situ hybridization performed with E3 chick embryos demonstrated that, in addition to the documented expression of Pax-QNR/Pax-6 in the neural tube, this gene is also expressed in the pancreatic bud. This expression is later restricted to discrete parts of the organ. From bacterially expressed Pax-QNR peptides we obtained rabbit antisera (paired domain, serum 11; domain between paired and homeo, serum 12; homeodomain, serum 13; and carboxyl-terminal part, serum 14) capable of specifically recognizing Pax-QNR/Pax-6 proteins (48, 46 kilodaltons) in cell lines derived from alpha- and beta-pancreatic cells, but not from exocrine derived cell lines. We conclude that Pax-QNR/Pax-6 represents another gene expressed both in the endocrine pancreas and neuro-ectodermic tissues.

[Impact of an intervention improving the food supply (excluding school meals) with educational support in middle and high schools]. / Impact d'une intervention sur l'offre alimentaire et sur les comportements alimentaires des adolescents (hors restauration scolaire).

Within the Nutrition, Prevention, and Health Program for children and teenagers in Aquitaine, an experimental intervention was implemented in 2007-2008 in the middle and high schools in Aquitaine (southwest France). This intervention aimed to improve the eating habits of adolescents, combining actions to improve the food supply sold during recreational times (remove/limit fat and sugar products sold and promote the sale of fruits and bread) and health education actions to make adolescents aware of the concept of nutritional balance and steer their choice towards recommended products. The aim of this paper was to evaluate the impact of the intervention on the eating behavior of adolescents and the food supply sold during recreational times in middle and high schools in Aquitaine. A survey was conducted before and after the implementation of the intervention in seven middle and high schools that have implemented actions (intervention group) and eight middle and high schools that have not implemented actions (control group). In these schools, 1602 adolescents answered the survey before and 1050 after the intervention (samples were independent because of the anonymity of responses). The impact of the intervention on the dietary behavior of teenagers was modeled using logistic regression adjusted on potential confounding variables (sex, age, and educational status). In multivariate analyses, the intervention was associated with more frequent daily intake of breakfast (OR=2.63; 95% CI [1.89; 3.66]) and lower intake of morning snacks (OR=0.66; 95% CI [0.48; 0.90]), higher consumption of starchy foods (OR=1.77; 95% CI [1.30; 2.42]), bread at breakfast, morning snacks, and a light afternoon meal (OR=1.43; 95% CI [1.07; 1.90]), and the food supply sold at recreational times (OR=1.34 95% CI [1.01; 1.78]). These results show that the "Improving food supply in middle and high schools associated with educational support actions" project led to the sales of recommended foods during recreational times and improved students' eating behavior. These results encourage partners to pursue these actions in all volunteer middle and high schools.

Rapid and sensitive method for the detection of B19 virus DNA using the polymerase chain reaction with nested primers.

The sensitivity of detection of B19 virus DNA in clinical specimens was evaluated by comparing the results of single PCR and nested PCR assays, with or without subsequent Southern blot hybridization to a radiolabelled B19 DNA probe. The inhibitory activity of human serum components on polymerase reaction was also determined. The sensitivity of B19 virus DNA detection decreased by a factor of 10(7) in the presence of 10% serum in the single PCR reaction mixture, and of 10(3) for nested PCR. When nested PCR products were analysed by Southern blot hybridization to a B19 radioactive DNA probe, the sensitivity of the assay increased to such a level of B19 DNA detection that the reaction was no longer influenced by the presence of serum inhibitors in the original sample. Less than ten B19 genome copies could thus be detected in a 10-microliters sample. A panel of 38 clinical samples, originating from patients with possibility of B19 virus infection, were assayed by this method. Only one sample was found to be positive after single PCR, whereas seven samples (including the former) gave a positive signal after nested PCR. The specificity of the nested PCR products was controlled by hybridization to the B19 DNA probe and DNA sequencing. No discrepancy in the results was observed between nested PCR alone and nested PCR followed by Southern blot analysis.

Implantable insulin pumps: infections most likely due to seeding from skin flora determine severe outcomes of pump-pocket seromas.

Complications at implantation site of implantable insulin pumps may lead to premature removal. To elucidate the origins and the outcomes of these local adverse events. We investigated seromas of the 'pump-pocket' that have been detected for an eight month-period during the follow-up of such-treated forty type 1 diabetic patients. At the start of study period, skin bacterial flora was sampled at umbilicus and groin, and isolated strains of Staphylococcus epidermidis were preserved in specific vials at -20 degrees C. Each time a seroma was detected at transcutaneous 45 days-refill of pump reservoir, it was sampled for bacterial cultures. Isolated strains of S. epidermidis from seroma were genetically compared to preserved strains of corresponding patients using Pulsed-Field Gel Electrophoresis (PFGE) after genomic restriction by SmaI. Among the ten seromas that occurred after a mean time of 9.9 months since implantation, S. epidermidis were isolated in five cases. Genetic comparison of isolated strains could be performed in three cases. Compared strains showed identical (in 2 cases) or closely related (in one case) PFGE profiles. While the five aseptic seromas resolved with rest, four infected cases required explantations after one to nineteen months in spite of antibiotic therapy and the fifth one persisted without impairment under long-term antibiotics. Our results suggest that seeding from the skin flora is a key-factor determining the severity of pump-pocket complications. We recommend that bacterial investigations of pump-pocket seromas should be systematically performed, while prophylactic measures might include antibiotic cover for each puncture of the pump-pocket.