The synthesis of unlabeled and 32P-labeled phosphocitrate and analytical systems for its identification

A method for the synthesis of phosphocitrate is described using 2-cyanoethyl phosphate to phosphorylate triethyl citrate. Following alkaline hydrolysis of the coupled intermediate, phosphocitrate was purified by ion-exchange chromatography on an AG 1-X8 (HCO₃^?) column. The method was also used to prepare [³²P]phosphocitrate. Phosphocitrate was characterized by ¹H NMR, ³¹P NMR, and ¹³C NMR spectroscopy. In addition methods for thin-layer chromatography and enzyme assay are detailed for the detection of phosphocitrate.     

Expression of N-linked sialyl Le(x) determinants and O-glycans in the carbohydrate moiety of human amniotic fluid transferrin during pregnancy

Transferrin, a glycoprotein involved in iron transport in body fluids, was isolated from amniotic fluid of a hydramniospatient by sequential anion-exchange chromatography and gel filtration. The N-glycans of human amniotic fluid transferrin (hAFT) were enzymatically liberated by PNGase-F digestion, isolated by gel filtration and fractionated by (high-pH) anion-exchange chromatography. After alkaline borohydride treatment of native hAFT, the released O-glycans were isolated by gel filtration and fractionated by anion-exchange chroma-tography. Structure elucidation of 14 N- and 2 O-glycans was performed by 500 or 600 MHz1H-NMR spectroscopy. Besides conventional N-glycans established earlier for human serum transferrin (hST), new (alpha1-3)-fucosylated N- glycans were found, representing sialyl Le(x) elements. Furthermore, as compared to hST, a higher degree of (alpha1-6)-fucosylation and an increase in branching from di- to triantennary compounds has been detected. The presence of O-glycans is demonstrated for the first time in transferrin.      

Imagerie et bilan d'une infertilité masculine

A male cause is responsible in near 50% of infertilities. The radiologist takes place in a multidisciplinary management, after clinical and biological screening, which distinguishes:

- excretory infertilities, of which some causes are curable. Transrectal sonography (TRUS) and scrotal sonography are the first tests. In case of epididymal obstacle, scrotal abnormalities may correspond to constitutional or acquired causes; TRUS is normal. TRUS usually identifies congenital bilateral absence of vas deferens; without renal agenesia, a genital form of cystic fibrosis must be evocated. In case of distal obstacles, TRUS may be completed with MRI, especially in case of voluminous cystic tumors. Vasography, which directly shows was deferens patency, is required to accurately diagnose obstruction when ultrasound is not conclusive; vasography must be integrated in a surgical strategy.

- secretory azoospermies, from gonadic or hypothalamo-hypophyseal causes. Scrotal sonography may complete clinical examination. When hypothalamo-hypophyseal axis must be explored, MRI is the reference test.

- oligo-astheno-teratospermies, where infertilities are often mixed, with various male factors.

Three groups must be explored: hyperprolactinemies (MRI); chronic genital infection (ultrasound); varicoceles; Doppler color ultrasound may help to the detection; spermatic phlebography produce a pretherapeutic cartography, and may be the first step of a percutaneous sclerotherapy.     

Effects of dipole position,orientation and noise on the accuracy of EEG source localization

Background  

The electroencephalogram (EEG) reflects the electrical activity in the brain on the surface of scalp. A major challenge in this field is the localization of sources in the brain responsible for eliciting the EEG signal measured at the scalp. In order to estimate the location of these sources, one must correctly model the sources, i.e., dipoles, as well as the volume conductor in which the resulting currents flow. In this study, we investigate the effects of dipole depth and orientation on source localization with varying sets of simulated random noise in 4 realistic head models.     

Molecular mechanism of ligand recognition by membrane transport protein,Mhp1

The hydantoin transporter Mhp1 is a sodium‐coupled secondary active transport protein of the nucleobase‐cation‐symport family and a member of the widespread 5‐helix inverted repeat superfamily of transporters. The structure of Mhp1 was previously solved in three different conformations providing insight into the molecular basis of the alternating access mechanism. Here, we elucidate detailed events of substrate binding, through a combination of crystallography, molecular dynamics, site‐directed mutagenesis, biochemical/biophysical assays, and the design and synthesis of novel ligands. We show precisely where 5‐substituted hydantoin substrates bind in an extended configuration at the interface of the bundle and hash domains. They are recognised through hydrogen bonds to the hydantoin moiety and the complementarity of the 5‐substituent for a hydrophobic pocket in the protein. Furthermore, we describe a novel structure of an intermediate state of the protein with the external thin gate locked open by an inhibitor, 5‐(2‐naphthylmethyl)‐L‐hydantoin, which becomes a substrate when leucine 363 is changed to an alanine. We deduce the molecular events that underlie acquisition and transport of a ligand by Mhp1.     

Active transportation to school over 2 years in relation to weight status and physical activity

Objective: To prospectively examine potential benefits of active commuting to school on measures of weight status and physical activity in a sample of youth. Research Methods and Procedures: A cohort of students from seven elementary schools was measured four times—in the fall and spring of fourth grade (N = 1083) and fifth grade (N = 924). Participants were classified as active (walking, biking, or skateboarding to school almost every day for baseline analyses or at least 2 d/wk for analyses of consistent active commuting) or non‐active commuters to school. Accelerometers were used to measure physical activity. Height, weight, and skinfolds were objectively assessed. Results: Boys who actively commuted to school had lower BMI (p < 0.01) and skinfolds (p < 0.05) than non‐active commuters to school in the fourth grade. Active commuting to school over 2 years was not associated with BMI change or overweight status. Discussion: Walking and cycling to school may contribute to preventing excessive weight gain, or leaner children may walk or cycle to school.     

SKPDB: a structural database of shikimate pathway enzymes

Background  

The functional and structural characterisation of enzymes that belong to microbial metabolic pathways is very important for structure-based drug design. The main interest in studying shikimate pathway enzymes involves the fact that they are essential for bacteria but do not occur in humans, making them selective targets for design of drugs that do not directly impact humans.