Hepatitis B virus in drug users in France: prevalence and vaccination history, ANRS-Coquelicot Survey 2011-2013.

People who use drugs (PWUD) are a key population for hepatitis B virus (HBV) vaccination and screening. We aimed to estimate the seroprevalence of HBs antigen (HBsAg) and self-reported HBV vaccination history in French PWUD attending harm reduction centres using data from the ANRS-Coquelicot multicentre survey conducted in 2011-2013 in 1718 PWUD. Self-fingerprick blood samples were collected on dried blood spots to detect the presence of HBsAg. HBsAg seroprevalence was estimated at 1·4% [95% confidence interval (CI) 0·8-2·5]. It varied between PWUD born in high (7·6%, 95% CI 2·7-19·1), moderate (2·2%, 95% CI 0·8-5·7) and low (0·7%, 95% CI 0·3-1·5) endemic zones. Factors independently associated with HBsAg carriage were being born in a moderate or high endemic zone or reporting precarious housing. Self-reported HBV vaccination history varied from 47·4% in high endemic zones, to 59·3% and 62·6% for moderate and low endemic zones, respectively. Our results suggest that drug use plays a small and substantial role, respectively, in HBsAg carriage in PWUD born in high/moderate and low endemic zones.

Pharmacokinetic interactions of OBE001 and betamethasone in healthy female volunteers.

WHAT IS KNOWN AND OBJECTIVE: To treat preterm labour, antenatal corticosteroids and tocolytics are often co-administered. OBE001 is an orally active oxytocin receptor antagonist under development for preterm labour treatment. METHODS: Co-administration of OBE001 and betamethasone to determine pharmacokinetic interactions was studied during an open-label, randomized, three-period crossover study. Twelve healthy post-menopausal volunteers received either two consecutive OBE001 administrations of 600 mg/day, two intramuscular injections of 12 mg/day betamethasone or the two drugs administered in combination. The area under the plasma concentration-time curve (AUC), the maximum plasma concentration (Cmax) and the time to Cmax (tmax) for OBE001 and betamethasone were measured. RESULTS AND DISCUSSION: There was no effect on geometric mean Cmax after the second administration and AUCs of OBE001 [geometric mean ratio point estimate (90% CI): Cmax (Day2) 1·05 (0·98-1·12) and AUC(0-24 h )1·11 (0·99-1·23)/AUC(24 h-∞) 0·99 (0·93-1·06), respectively]; Cmax after the first administration together with betamethasone was increased by 12% [geometric mean ratio point estimates (90% CI): Cmax (Day1) 1·12 (0·96-1·32)]. Tmax after concomitant administration with betamethasone occurred with a median delay of 1 h. Geometric mean Cmax and AUCs of betamethasone were not affected by concomitant OBE001 administration [geometric mean ratio point estimate (90% CI): Cmax (Day1) 1·02 (0·98-1·07)/Cmax (Day2) 1·03 (0·98-1·08) and AUC(0-24 h) 1·07 (1·04-1·11)/AUC(24 h-∞ )1·04 (1·01-1·08), respectively], with no effect on median tmax . No subject was discontinued from the study due to adverse events. WHAT IS NEW AND CONCLUSION: AUC and Cmax of the betamethasone and OBE001 combination did not differ significantly between treatments. Co-administration of OBE001 and betamethasone was well tolerated and resulted in a tmax median delay of 1 h for OBE001 but not for betamethasone. Co-administration of OBE001 and betamethasone in clinics is feasible and does not require any specific precaution or administration adaptation.

Maternal tobacco smoking in pregnancy and children's socio-emotional development at age 5: The EDEN mother-child birth cohort study.

BACKGROUND: There is debate as to whether maternal tobacco use in pregnancy is related to offspring behaviour later on. We tested this association examining multiple aspects of children's behaviour at age 5 and accounting for parental smoking outside of pregnancy, as well as child and family characteristics. METHODS: Data come from a prospective community based birth cohort study (EDEN; n=1113 families in France followed since pregnancy in 2003-2005 until the child's 5th birthday). Maternal tobacco use in pregnancy was self-reported. Children's socio-emotional development (emotional symptoms, conduct problems, symptoms of hyperactivity/inattention, peer relationship problems, prosocial behaviour) was assessed by mothers using the Strengths and Difficulties Questionnaire (SDQ) at age 5 years. Logistic regression analyses controlled for Inverse Probability Weights (IPW) of maternal tobacco use calculated based on study center, children's characteristics (sex, premature birth, low birth weight, breastfeeding), maternal characteristics (age at the child's birth, psychological difficulties and alcohol use in pregnancy, post-pregnancy depression, and smoking), paternal smoking in and post-pregnancy, parental educational attainment, family income, parental separation, and maternal negative life events. RESULTS: Maternal smoking in pregnancy only predicted children's high symptoms of hyperactivity/inattention (sex and study center-adjusted ORs: maternal smoking in the 1st trimester: 1.95, 95%CI: 1.13-3.38; maternal smoking throughout pregnancy: OR=2.11, 95%CI: 1.36-3.27). In IPW-controlled regression models, only children of mothers who smoked throughout pregnancy had significantly elevated levels of hyperactivity/inattention (OR=2.20, 95%CI: 1.21-4.00). CONCLUSIONS: Maternal tobacco smoking in pregnancy may contribute directly or through epigenetic mechanisms to children's symptoms of hyperactivity/inattention.

Calcium influx and protein kinase C alpha activation mediate arachidonic acid mobilization by the human NK-2 receptor expressed in Chinese hamster ovary cells.

We have cloned a cDNA encoding the human ileal neurokinin-2 (NK-2) receptor which mediates powerful neurokinin A-stimulated arachidonic acid (AA) and prostaglandin release when expressed in CHO cells. Two major signal transduction events appear to underlie this response. Firstly, AA liberation is critically dependent upon agonist-stimulated influx of extracellular Ca2+ although not release from intracellular stores. Secondly, NK-2 receptor-linked AA mobilization requires concomitant PKC activation and based upon limited subtype immunodetectability as well as toxin, identical pretreatment inhibits AA release partially and blocks PKC alpha translocation completely. These observations indicate that in this cell system AA liberation reflects NK-2 receptor-dependent activation of two distinct but converging signal transduction pathways regulated by different G-protein species and involving Ca2+ influx and PKC alpha activation.

Palmitoylation but not the extreme amino-terminus of Gq alpha is required for coupling to the NK2 receptor.

Gq alpha and G11 alpha differ from other G protein alpha subunits in that they have unique, conserved 6 residue amino-terminal extensions. Wild-type and amino-terminal mutants of Gq alpha expressed in COS cells were analyzed for their ability to functionally couple with co-expressed neurokinin NK2 receptor. Wild-type, T2A and delta 2-7 Gq alpha were able to stimulate agonist driven phospholipase C (PLC) activity in identical manners. Other activities of these two amino-terminal mutants including aluminum fluoride stimulated PLC activity, palmitoylation, interaction with G beta gamma subunits and GTP gamma S-induced trypsin resistance are also similar to the wild-type alpha subunit. This demonstrates that the NK2 receptor is able to functionally interact with the alpha subunit of Gq and that the first seven amino-acids of Gq alpha are not required for any of the alpha subunit functions tested. In contrast to the T2A and delta 2-7 mutants, a C9,10A Gq alpha mutant was not able to couple to either the NK2 receptor or PLC, as assessed by high-affinity agonist binding and activation of PLC either in intact cells or in vitro. The C9,10A protein was able to assume a GTP gamma S-induced trypsin-resistant conformation and partitioned primarily to the pelletable fraction in a manner similar to the wild-type protein. However, it was not labeled with [3H]palmitic acid. This suggests that blocking palmitoylation at the amino-terminus of Gq alpha results in a loss of functional activity which reflects an inability to interact with both the receptor and downstream signaling targets.

Expression and characterization of human D4 dopamine receptors in baculovirus-infected insect cells.

The human D4 dopamine receptor has been genetically engineered for expression in insect cells using the baculovirus system. A D4 cDNA gene fusion construct [(1991) Nature 350, 610-614] was synthetically modified to remove two introns from the coding region, and expressed in S. frugiperda (Sf9) cells as a fusion with a short sequence from the polyhedrin protein. Binding assays with [3H]spiperone indicated high levels of D4 receptor binding 90 h after infection and a pharmacological profile identical to that reported for D4 receptors expressed in COS-7 cells using the cDNA gene hybrid. We also show that the agonist binding affinity of D4 receptors expressed in Sf9 cells can be shifted by GTP-gamma-S, indicating coupling to G-proteins.

Preparation, application and biological characterization of interleukin-1 beta mutant protein biotinylated at a single site.

Selective biotinylation of human interleukin-1 beta was achieved by reaction of an interleukin-1 beta mutant protein containing a surface-accessible cysteine with maleimido-biotin. A defined interleukin-1 beta derivative biotinylated at a single site was obtained. This compound retained full affinity for both the interleukin-1 receptor and streptavidin or avidin; however, its biological activity was significantly reduced in proliferative assays. Biotinylated interleukin-1 beta mutant protein bound to fluorescein-labelled avidin was used in flow cytometric analysis of interleukin-1 receptors. Internalization of the complex between biotinylated interleukin-1 beta mutant protein and streptavidin in a murine thymoma cell line was demonstrated.

Synthesis and characterization of fluorescent and photoactivatable MIP-1alpha ligands and interactions with chemokine receptors CCR1 and CCR5.

Photoaffinity and fluorescent analogues of the 70-amino acid chemokine macrophage inflammatory protein-1alpha (MIP-1alpha) were designed, synthesized, characterized, and applied to probe MIP-1alpha interactions with the chemokine receptors CCR1 and CCR5. The photoactivatable MIP-1alpha ligand, BP-MIP-1alpha, and the fluorescent ligand, Flu-MIP-1alpha were prepared by selective chemical coupling of p-benzoylphenylthiocarbamyl or fluoresceinthiocarbamyl, respectively, at the N-terminus of MIP-1alpha. Both ligands BP-MIP-1alpha and Flu-MIP-1alpha retained high binding affinity and agonist potency at CCR1 and CCR5. Photoaffinity labeling of CCR1 and CCR5 receptors stably expressed in CHO cells resulted in specific covalent attachment of [(125)I]BP-MIP-1alpha and production of protein complexes of 54 and 48 kDa, respectively, on SDS-PAGE. This represents the first photo-cross-linking between a chemokine and its receptor. Flu-MIP-1alpha selectively labeled CCR1 or CCR5 receptors expressed in CHO cells and was used to characterize receptor binding domains. When bound to CCR1 or CCR5 receptors, the fluorescence signal of Flu-MIP-1alpha was quenched by collision with iodide indicating that the N-terminal end of MIP-1alpha is accessible to the solvent. These data strongly suggest that the N-terminal end of MIP-1alpha interacts with domains of CCR1 or CCR5 receptors located at the extracellular surface. The photoactivatable BP-MIP-1alpha described here should prove valuable for the identification of contact sites on receptors by photoaffinity labeling experiments.

Synthesis and characterization of selective fluorescent ligands for the neurokinin NK2 receptor.

Several fluorescent probes for the NK2 receptor were designed, synthesized, and pharmacologically characterized. These fluorescent ligands are analogues of the selective NK2 heptapeptide antagonist N-alpha-benzoyl-Ala-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH2 (1, GR94800). They were obtained by substitution of 2,n-diaminoalkyl amino acid (n = 3-6) for Ala1 and the subsequent coupling of the fluorophore NBD (7-nitrobenz-2-oxa-1,3-diazol-4-yl) or fluoresceinthiocarbamyl to the N-omega amino group. The fluorescent derivatives retained high binding affinities for the NK2 receptor in transfected CHO cells. In contrast, fluorescent derivatives made by replacing the N-alpha-benzoyl group of 1 by NBD or fluorescein were considerably less active. The effect on ligand potency of varying the length of the spacer arm between the peptide moiety and the fluorescent group was also studied. The most potent fluorescent antagonists were N-alpha-benzoyl-Dab(gamma-NBD)-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH2 (5B), pKi = 8.87 for NK2; N-alpha-benzoyl-Orn (delta-NBD)-Ala-D-Trp-Phe- D-Pro-Pro-Nle-NH2 (4B), pKi = 8.84; and N-alpha-benzoyl-Lys(epsilon-NBD)-Ala-D-Trp-Phe-D-Pro-Pro-Nle-NH2 (3B), pKi = 8.83. These three compounds were highly selective for NK2 over NK3 and NK1 receptors. We show that these fluorescent ligands are useful tools for the detection of NK2 receptor expression by flow cytometry. Additionally, these fluorescent probes should prove valuable for fluorescence microscopy and study of ligand-receptor interaction by spectrofluorimetry.

Fluorescent pseudo-peptide linear vasopressin antagonists: design, synthesis, and applications.

Fluoresceinyl and rhodamyl groups have been coupled by an amide link to side-chain amino groups at positions 1, 6, and 8 of pseudo-peptide linear vasopressin antagonists (Manning et al. Int. J. Pept. Protein Res. 1992, 40, 261-267) through different positions on the fluorophore, to give tetraethylrhodamyl-DTyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-Tyr-NH2 (2), 4-HOPh(CH2)2CO-DTyr(Me)-Phe-Gln-Asn-Lys(5-carboxyfl uoresceinyl)-Pro-A rg-NH2 (4), 4-HOPh(CH2)2CO-DTyr(Me)-Phe-Gln-Asn-Lys(5- or 6-carboxytetramethylrhodamyl)-Pro-Arg-NH2 (5, 6), 4-HOPh(CH2)2CO-DTyr(Me)-Phe-Gln-Asn-Arg-Pro-Lys(5- or 6- carboxyfluoresceinyl)-NH2 (8, 9), and 4-HOPh(CH2)2CO-DTyr(Me)-Phe-Gln-Asn-Arg-Pro-Lys(5- or 6- carboxytetramethylrhodamyl)-NH2 (10, 11). The closer to the C-terminus the fluorophore, the higher the affinities of the fluorescent derivatives for the human vasopressin V1a receptor transfected in CHO cells. The compound 10 has a Ki of 70 pM, as determined by competition experiments with [125I]-4-HOPhCH2CO-DTyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-NH2. It showed a good selectivity for human V1a receptor versus human OT (Ki = 1.2 nM), human vasopressin V1b (Ki approximately 27 nM), and human vasopressin V2 (Ki > 5000 nM) receptor subtypes. All fluorescent analogues were antagonists as shown by the inhibition of vasopressin induced inositol phosphate accumulation. These fluorescent ligands are efficient for labeling cells expressing the human V1a receptor subtype, as shown by flow cytofluorometric experiments or fluorescence microscopy. They are also appropriate tools for structural analysis of the vasopressin receptors by fluorescence.

An asanguineous reperfusion solution. An effective adjunct to cardioplegic protection in high-risk valve operations.

The protection afforded by cardioplegia during elective ischemic arrest can be partly compromised by a reperfusion injury, which may impede the recovery of cardiac function. We previously showed experimentally that this postischemic damage could be largely avoided by an appropriate crystalloid reperfusate. The present study was thus undertaken to assess the effects of this "reperfusion solution" clinically. One hundred twelve patients undergoing valve replacement with the aid of hypothermic cardioplegia (K+ 12 mEq, Mg2+ 26 mEq) were prospectively divided in two groups: Group I (n = 49) received an unmodified blood reperfusate. In Group II (n = 63), 1 L of the reperfusion solution was delivered just prior to removal of the aortic clamp. The formulation of the reperfusion solution adhered to the following principles: (1) maintenance of cardioplegia (K+ = 15 mEq), (2) replenishment of Ca2+ stores (Ca2+ = 2.5 mEq), (3) substrate provision (glutamate = 2,942 gm), (4) buffering (pH = 7.70 at 28 degrees C), and (5) hyperosmolarity (370 mOsm). The two groups were matched for preoperative data except for a higher incidence of isolated aortic valve replacement (p = 0.01) in Group II. Also, the cross-clamp time (mean +/- standard error of the mean) was longer in Group II (94 +/- 4 minutes versus 63 +/- 4 minutes, p less than 10(-6]. The reperfusion solution was found to increase both the rate and extent of postischemic functional recovery, as evidenced by (1) a lower proportion of catecholamine-supported patients 48 hours after operation (9/63 [14.28%] versus 16/49 [32.6%] in the control group [p less than 0.03]) and (2) a lower amount (gamma/kg/min) of dobutamine required to achieve stable hemodynamics (11 +/- 1 versus 26 +/- 6 in the control group [p less than 0.03]). A similar recovery pattern was noted in the high-risk subgroup of patients with mitral valve disease. Further, serial postoperative hemodynamic measurements were performed in 31 randomly selected patients (10 control and 21 reperfused). Although the reperfused patients were found to be at higher risk because of lower preoperative cardiac indices and longer cross-clamp times, they consistently achieved better postoperative hemodynamics with a lower incidence of catecholamine support. This hemodynamic improvement was particularly reflected by a higher left ventricular stroke work index throughout the postoperative course, the difference being significant 6 hours and 12 hours postoperatively.(ABSTRACT TRUNCATED AT 400 WORDS)

The lateral forearm flap: an anatomic study.

The anatomy of the lateral forearm flap has been studied in 12 fresh cadaver arms with methylene blue and latex injections and arteriography. The posterior radial collateral artery was found to divide constantly into two terminal branches, an anterior and a posterior division. The anterior division is the nutrient vessel of the flap. This artery extends significantly beyond the lateral epicondyle of the elbow into the lateral aspect of the forearm (range 13 to 18 cm, average 15 cm). This allows raising a fasciocutaneous flap in the proximal forearm with a much longer vascular pedicle than the classic lateral arm flap. Other advantages include very thin skin and subcutaneous tissue and less sensory deficit at the donor site. Based on these results, this newly designed lateral forearm flap has been used in 13 clinical cases. Its main indications are whenever soft, thin, pliable skin is needed for small to moderate-sized defects.

Periosteal suspension of the lower eyelid and cheek following subciliary exposure of facial fractures.

Following craniofacial procedures that involve stripping of the periosteum and soft tissue over the zygomatic maxillary complex, descent of soft tissue with a decrease in anterior projection over the malar area and increase in fullness in the nasolabial fold have been seen to be a problem by these authors. Simple repositioning of the soft tissues to their normal anatomic position may be used to alleviate this problem.

Emergency reconstruction of a collateral ligament of a metacarpophalangeal joint using Dacron material.

We present a case in which an open wound involving the ulnar collateral ligament of the metacarpophalangeal joint of the little finger was treated by ligament reconstruction using a strip of Dacron material, nerve grafting and coverage by a posterior interosseous artery pedicled flap. At a long term follow-up of 4 years, the joint was stable and had a full range of movement.

Versatility of the homodigital triangular neurovascular island flap in fingertip reconstruction.

This paper reports 25 cases of a versatile homodigital triangular neurovascular island flap for fingertip reconstruction. This flap was used to reconstruct traumatic oblique palmar amputations, with loss of fingertip pulp requiring advancement of not more than 2 cm. Good results were obtained in terms of fingertip contour and padding, sensibility, and functional recovery. The tendency for flexion contracture at the PIP joint was prevented by using a dynamic extension splint at night. Cold intolerance must be expected, especially in cold countries.

[Fatal hypoxemia following extracorporeal circulation for aortocoronary bypass. A case]. / Hypoxémie mortelle après circulation extra-corporelle pour pontage aorto-coronarien. Une observation.

A lethal case of acute respiratory distress syndrome of the adult following extracorporeal circulation for aorto-coronary bypass is reported. This case illustrates the two mechanisms currently thought to be responsible for this pathology: (1) the passage of blood over the synthetic fibres of the cardiopulmonary bypass machine activates the complement, and the systemic inflammatory reaction that ensues accounts for the increased permeability of the alveolar capillary membrane; (2) the haemodilution inherent in extracorporeal circulation explains the hypoproteinaemia usually observed at the end of the procedure; the resulting fall in capillary osmotic pressure upsets the equilibrium of pressures in the capillary vessels.

Isoniazid: an update on the multiple mechanisms for a singular action.

Isoniazid (INH) is one of the most commonly used drugs in treatment of human tuberculosis and the most efficient. Although it has been 60 years since isoniazid was introduced in anti-tubercular therapy and despite the simplicity of its chemical structure (C6H7N3O) with few functional groups, its exact mechanism of action, which could account for its specificity and exceptional potency against Mycobacterium tuberculosis and justify all profiles of INH-resistance, remains elusive and debatable. This complexity can find an explanation in the high reactivity of INH and also in the possibility that multiple targets and pathways could co-exist for this medicinal agent. Indeed, since the discovery of isoniazid's anti-tubercular potency, several propositions for its mode of action have been reported, including its conversion, by a catalase peroxidase within M. tuberculosis, into an active metabolite able, after reaction with NAD, to inhibit an enzyme (InhA) crucial to M. tuberculosis survival. This represents the most consensual mechanism described to date. Nevertheless, none of the proposed mechanisms considered independently can explain the singular and privileged action of the isoniazid structure on the tubercle bacillus, or all the profiles of resistance. The aim of this paper is to reconsider the literature reporting the different modes of action described for isoniazid in the light of the present and most relevant knowledge, with special attention to understanding the molecular mechanistic aspects of the drug's action.

[Setting up supportive care in oncology: reflexions and suggestions.]. / A propos de la mise en place des soins de support en cancérologie : pistes de réflexions et propositions.

A group of 19 health professionals implicated in supportive care wanted to suggest some reflexions for organization, setting and evaluation of the supportive care in institutions and health territories. The suggested organization must be applicable to any cancer patient and the place of the care whatever the age, the stage of the disease; in the future, must be applicable to any patient with serious chronic illness. This organization must allow to optimize the accompaniment and the care of the patients and their close relations by 1) precise and regular analysis of their needs; 2) the respect of the continuity of the health care; 3) the setting of collaborative practice and transversality in the care. It is not a new medical speciality but a coordination of competences for patients and their families.