A simplified method for the detection of Y chromosome microdeletions in infertile men using a multiplex sequence-tagged site-based amplification

Sixteen sequence-tagged sites (STSs) were combined in five amplification reactions, to screen for deletions of DNA fragments located within the AZFa, AZFb, and AZFc regions of the Y chromosome. This multiplex strategy is fast and reliable, and most of the azoospermia-associated deletions reported so far are detected with this simplified method. Internal control STSs are included that allow discrimination between deletion and failure of amplification.     

Improved absorption of caseinophosphopeptide-bound iron: role of alkaline phosphatase

Hydrolysis of proteins could lessen their inhibiting effect on the poor absorption of cow's milk iron (Fe), which is responsible for the high incidence of Fe deficiency worldwide. When bound to Fe, caseinophosphopeptides (CPP) derived from milk proteins resist luminal digestion, enhance Fe solubility and could improve its bioavailability; brush border enzyme alkaline phosphatase activity could influence iron absorption by releasing free Fe; this study assessed its role in the absorption of CPP-bound Fe. Rat duodenal loops were perfused with Fe gluconate or Fe bound to the CPP of beta casein [beta-CN (1-25)], with or without the addition of an inhibitor of alkaline phosphatase, Na2WO4. The uptake of Fe-beta-CN (1-25) was greater than Fe gluconate. Na2WO4 enhanced the uptake of Fe-beta-CN (1-25) and not of Fe gluconate. So the release of free, insoluble Fe, by alkaline phosphatase seems to be prevented by providing Fe in the Fe-beta-CN (1-25) complex form. Its good disappearance rate makes beta-CN (1-25)-bound Fe a candidate for food fortification.     

Symptomatic Management of Fever in Children: A National Survey of Healthcare Professionals’ Practices in France

Despite the production and dissemination of recommendations related to managing fever in children, this symptom saturates the practices of primary healthcare professionals (HPs). Data on parent practices related to fever are available, but data on HPs’ practices are limited. We studied HPs’ practices, determinants of practices and concordance with recommendations in France. We conducted a national cross-sectional observational study between 2007 and 2008 among French general practitioners, primary care pediatricians and pharmacists. HPs were asked to include 5 consecutive patients aged 1 month to 12 years with acute fever. HPs completed a questionnaire about their practices for the current fever episode. We used a multilevel logistic regression model to assess the joint effects of patient- and HP-level variables associated with this behavior. In all, 1,534 HPs (participation rate 13%) included 6,596 children (mean age 3.7 ± 2.7 years). Physicians measured the temperature of 40% of children. Primary HPs recommended drug treatment for 84% of children (including monotherapy for 92%) and physical treatment for 62% (including all recommended physical treatments for 7%). HPs gave written advice or a pamphlet for 13% of children. Significant practice variations were associated with characteristics of the child (age, fever level and diagnosis) and HP (profession and experience). In France, despite the production and dissemination of national recommendations for managing fever in children, primary HPs’ observed practices differed greatly from current recommendations, which suggests potential targets for continuing medical education.     

Managing Fever in Children: A National Survey of Parents' Knowledge and Practices in France

Introduction

Identifying targets to improve parental practices for managing fever in children is the first step to reducing the overloaded healthcare system related to this common symptom. We aimed to study parents'' knowledge and practices and their determinants in managing fever symptoms in children in France as compared with current recommendations.

Methods

We conducted an observational national study between 2007 and 2008 of French general practitioners, primary care pediatricians and pharmacists. These healthcare professionals (HPs) were asked to include 5 consecutive patients from 1 month to 12 years old with fever for up to 48 hr who were accompanied by a family member. Parents completed a questionnaire about their knowledge of fever in children and their attitudes about the current fever episode. We used a multilevel logistic regression model to assess the joint effects of patient- and HP-level variables.

Results

In all, 1,534 HPs (participation rate 13%) included 6,596 children. Parental concordance with current recommendations for temperature measurement methods, the threshold for defining fever, and physical (oral hydration, undressing, room temperature) and drug treatment was 89%, 61%, 15%, and 23%, respectively. Multivariate multi-level analyses revealed a significant HP effect. In general, high concordance with recommendations was associated with high educational level of parents and the HP consulted being a pediatrician.

Conclusions

In France, parents'' knowledge and practices related to managing fever symptoms in children frequently differ from recommendations. Targeted health education interventions are needed to effectively manage fever symptoms in children.     

Comparative capacities of the pig colon and duodenum for luminal iron absorption

Iron deficiency is the most common human nutritional disorder in the world. Iron absorptive capacity of the small intestine is known to be much limited and therefore large quantities of iron salts must be used to treat iron deficiency. As a result, significant amounts of iron may reach the large intestine. This study compared the capacities of the small and large intestine to transfer luminal iron to the venous blood in relationship with the expression in epithelial cells of proteins involved in iron absorption using a pig model. Intracaecal injection of iron sulphate corresponding with 2.5 and 5.0 mg elemental iron per kg body mass resulted in modest, transient, but significant (p<0.05) increases in iron concentration in the portal blood plasma. By comparing portal blood plasma iron concentrations following injection in the duodenal and caecal lumen, we calculated that 5 h after injection, iron colonic absorption represented approximately 14% of duodenal absorption. Caecal and proximal colon mucosa accumulated iron to a much lower extent than the duodenal mucosa. Isolated colonocytes were found to express divalent metal transporter (DMT1) and ferritin, but to a lesser extent than the duodenal enterocytes. Ferroportin was highly expressed in colonocytes. In these cells as well as in enterocytes ferroportin was found to be glycosylated. In short term experiments and at a concentration in the range of that measured in the aqueous phases recovered from the large intestine luminal content after iron injection, iron sulphate did not alter colonocyte viability. We concluded that the colonic epithelial cells that express proteins involved in iron absorption are able to transfer luminal iron to the venous blood even if its relative participation in the overall intestinal absorption appears to be modest under our experimental conditions.     

Fourier transform infrared spectroscopy as a new tool for characterization of mollicutes

Fourier transform infrared (FT-IR) spectroscopy is a convenient physico-chemical technique to investigate various cell materials. Bacteria of class Mollicutes, identified by conventional methods, as Mycoplasma, Acholeplasma and Ureaplasma genera were characterized using this method. A data set of 74 independent experiments corresponding to fourteen reference strains of Mollicutes was examined by FT-IR spectroscopy to attempt a spectral characterization based on the biomolecular structures. In addition to the separation of Mollicutes within the lipidic region into five main clusters corresponding to the three phylogenetic groups tested, FT-IR spectroscopy allowed a fine discrimination between strains belonging to the same species by using selective spectral windows, particularly in the 1200-900 cm(-1) saccharide range. The results obtained by FT-IR were in good agreement with both taxonomic and phylogenetic classifications of tested strains. Thus, this technique appears to be a useful tool and an accurate mean for a rapid characterization of Mollicutes observed in humans.     

MLVA subtyping of genovar E Chlamydia trachomatis individualizes the Swedish variant and anorectal isolates from men who have sex with men

This study describes a new multilocus variable number tandem-repeat (VNTR) analysis (MLVA) typing system for the discrimination of Chlamydia trachomatis genovar D to K isolates or specimens. We focused our MLVA scheme on genovar E which predominates in most populations worldwide. This system does not require culture and therefore can be performed directly on DNA extracted from positive clinical specimens. Our method was based on GeneScan analysis of five VNTR loci labelled with fluorescent dyes by multiplex PCR and capillary electrophoresis. This MLVA, called MLVA-5, was applied to a collection of 220 genovar E and 94 non-E genovar C. trachomatis isolates and specimens obtained from 251 patients and resulted in 38 MLVA-5 types. The genetic stability of the MLVA-5 scheme was assessed for results obtained both in vitro by serial passage culturing and in vivo using concomitant and sequential isolates and specimens. All anorectal genovar E isolates from men who have sex with men exhibited the same MLVA-5 type, suggesting clonal spread. In the same way, we confirmed the clonal origin of the Swedish new variant of C. trachomatis. The MLVA-5 assay was compared to three other molecular typing methods, ompA gene sequencing, multilocus sequence typing (MLST) and a previous MLVA method called MLVA-3, on 43 genovar E isolates. The discriminatory index was 0.913 for MLVA-5, 0.860 for MLST and 0.622 for MLVA-3. Among all of these genotyping methods, MLVA-5 displayed the highest discriminatory power and does not require a time-consuming sequencing step. The results indicate that MLVA-5 enables high-resolution molecular epidemiological characterisation of C. trachomatis genovars D to K infections directly from specimens.