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BackgroundCongenital toxoplasmosis is a treatable, preventable disease, but untreated causes death, prematurity, loss of sight, cognition and motor function, and substantial costs worldwide.Methods/FindingsIn our ongoing USA feasibility/efficacy clinical trial, data collated with other ongoing and earlier published results proved high performance of an Immunochromatographic-test(ICT) that enables accurate, rapid diagnosis/treatment, establishing new paradigms for care. Overall results from patient blood and/or serum samples tested with ICT compared with gold-standard-predicate-test results found ICT performance for 4606 sera/1876 blood, 99.3%/97.5% sensitive and 98.9%/99.7% specific. However, in the clinical trial the FDA-cleared-predicate test initially caused practical, costly problems due to false-positive-IgM results. For 58 persons, 3/43 seronegative and 2/15 chronically infected persons had false positive IgM predicate tests. This caused substantial anxiety, concerns, and required costly, delayed confirmation in reference centers. Absence of false positive ICT results contributes to solutions: Lyon and Paris France and USA Reference laboratories frequently receive sera with erroneously positive local laboratory IgM results impeding patient care. Therefore, thirty-two such sera referred to Lyon’s Reference laboratory were ICT-tested. We collated these with other earlier/ongoing results: 132 of 137 USA or French persons had false positive local laboratory IgM results identified correctly as negative by ICT. Five false positive ICT results in Tunisia and Marseille, France, emphasize need to confirm positive ICT results with Sabin-Feldman-Dye-test or western blot. Separate studies demonstrated high performance in detecting acute infections, meeting FDA, CLIA, WHO ASSURED, CEMark criteria and patient and physician satisfaction with monthly-gestational-ICT-screening.Conclusions/SignificanceThis novel paradigm using ICT identifies likely false positives or raises suspicion that a result is truly positive, rapidly needing prompt follow up and treatment. Thus, ICT enables well-accepted gestational screening programs that facilitate rapid treatment saving lives, sight, cognition and motor function. This reduces anxiety, delays, work, and cost at point-of-care and clinical laboratories.Author’s SummaryToxoplasmosis is a major health burden for developed and developing countries, causing damage to eyes and brain, loss of life and substantial societal costs. Prompt diagnosis in gestational screening programs enables treatment, thereby relieving suffering, and leading to > 14-fold cost savings for care. Herein, we demonstrate that using an ICT that meets WHO ASSURED-criteria identifying persons with/without antibody toToxoplasma gondiiin sera and whole blood with high sensitivity and specificity, is feasible to use in USA clinical practice. We find this new approach can help to obviate the problem of detection of false positive anti-T.gondiiIgM results for those without IgG antibodies toT.gondiiwhen this occurs in present, standard of care, predicate USA FDA cleared available assays. Thus, this accurate test facilitates gestational screening programs and a global initiative to diagnose and thereby prevent and treatT.gondiiinfection. This minimizes likelihood of false positives (IgG and/or IgM) while maintaining maximum sensitivity. When isolated IgM antibodies are detected, it is necessary to confirm and when indicated continue follow up testing in ∼2 weeks to establish seroconversion. Presence of a positive ICT makes it likely that IgM is truly positive and a negative ICT makes it likely that IgM will be a false positive without infection. These results create a new, enthusiastically-accepted, precise paradigm for rapid diagnosis and validation of results with a second-line test. This helps eliminate alarm and anxiety about false-positive results, while expediting needed treatment for true positive results and providing back up distinguishing false positive tests.

International audience; The burden of congenital toxoplasmosis has become small in France today, in particular as a result of timely therapy for pregnant women, fetuses and newborns. Thus, the French screening and prevention program has been evaluated and recently confirmed despite a decline over time in the incidence of toxoplasmosis. Serological diagnosis of maternal seroconversion is usually simple but can be difficult when the first trimester test shows the presence of IgM, requiring referral to an expert laboratory. Woman with confirmed seroconversion should be referred quickly to an expert center, which will decide with her on treatment and antenatal diagnosis. Although the level of proof is moderate, there is a body of evidence in favor of active prophylactic prenatal treatment started as early as possible (ideally within 3 weeks of seroconversion) to reduce the risk of maternal-fetal transmission, as well as symptoms in children. The recommended therapies to prevent maternal-fetal transmission are: (1) spiramycin in case of maternal infection before 14 gestational weeks; (2) pyrimethamine and sulfadiazine (P-S) with folinic acid in case of maternal infection at 14 WG or more. Amniocentesis is recommended to guide prenatal and neonatal care. If fetal infection is diagnosed by PCR on amniotic fluid, therapy with P-S should be initiated as early as possible or continued in order reduce the risk of damage to the brain or eyes. Further research is required to validate new approaches to preventing congenital toxoplasmosis.

Flow regulators are widely used in hospitals to assist with intravenous (IV) infusion of medication. The rupture of a flow regulator at the base of the clamp was observed during parenteral nutrition. This rupture resulted in fluid leakage and an inlet of air, responsible for an air embolism in a fragile patient who had undergone a bilateral lung transplant. The patient's clinical condition required him to be transferred to a continuous monitoring unit. A serious Adverse Event in Healthcare (AEH) was reported, as well as a medical device vigilance report. A Feedback Committee (FC) was set up and it recommended an audit within the health care departments to study the conditions for use of flow regulators and to propose corrective actions. Despite the technical data sheet of the device not recommending the administration of lipid emulsions and glucose solutions above 10%, the manufacturer's expert report concluded that the mechanical failure could not be linked to the type of solution. However, the audit did reveal a lack of knowledge of certain rules for using this device. The analysis of this AEH is part of the establishment's patient safety procedure. The AEH highlighted a deviation in care concerning the conditions for use of flow regulators, thus resulting in misuse. The collaboration between the various actors involved in the analysis of this AEH led to the implementation of improvement actions on the root causes, related to the lack of information and of training for professionals on correct use of the medical device.

Background and importance Despite a strict regulatory framework of clinical trials (CTs), few standardised tools are available. Our national survey conducted in 2020 demonstrated that all clinical research pharmacists (CRPs) have initiated a quality approach, which is however very heterogeneous and implemented more in university hospital centres and cancer centres, with a high activity level. New standardised tools for the investigational health products (IHPs) circuit would be of interest to 88/94 CRPs. The most useful tool was the self-assessment grid, according to 94% of CRPs. Thus we developed such a tool to manage specific risks of IHPs (complex protocols, assignment of treatment numbers, confusing packaging or labelling). Aim and objectives The aim of this work was create a standardised self-assessment grid to manage the specific risks of IHPs. Material and methods A pharmacy resident and two doctors of pharmacy of a regional working group defined a list of 66 evaluation criteria, mainly based on good clinical practices and on a professional guide of 2020 by the national university centre hospitals’ pharmacists commission. The criteria were divided into three main parts: general organisation and support functions; pharmaceutical management of CTs; and risk assessment and risk management. Then the grid was sent for validation to the 94 CRPs who had answered our national survey. The Delphi method was used and consensus among experts was defined by a satisfaction rate of >80% for relevance, clearness and accessibility. Results The first round of proofreading by 16 pharmacists led to a consensus of 85% (56/66) for the criteria, which were considered relevant, clear and assessable. This led to 36 modifications, 4 deletions and 2 additions of criteria. The second round by 8 pharmacists led to a consensus of 88% (60/68) for the criteria and resulted in 18 modifications and 2 deletions. There remained only one criterion considered irrelevant, which was deleted. These results led us to validate the final version of the grid, including 62 criteria. Conclusion and relevance This interactive tool will be disseminated in a free public online ‘CTs’ toolbox. It will provide a conformity score per process, allowing specific risks to be identified across the circuit of IHPs, by pharmacies in any healthcare facility, whatever the level of activity. References and/or acknowledgements Conflict of interest No conflict of interest

La toxoplasmosis es una infeccion parasitaria cosmopolita que afecta a entre un tercio y un cuarto de la poblacion mundial. Las fuentes de contaminacion multiples (carne, agua, tierra, verduras) explican la adquisicion a menudo precoz de la infeccion en el transcurso de la vida. La infeccion adquirida suele ser benigna y asintomatica, aunque puede producir linfadenopatias. En Francia, por ejemplo, la toxoplasmosis congenita adquirida tras la infeccion materna durante el embarazo afecta a alrededor de 300 ninos cada ano. Al nacer, mas del 80% de ellos no tiene signos clinicos, pero alrededor de un 25% desarrolla coriorretinitis. Las formas graves de toxoplasmosis congenita, neurooculares o diseminadas, son infrecuentes en Francia. Las lesiones oculares, sean adquiridas o congenitas, son mas graves y mas precoces en otros paises, sobre todo de America del Sur, debido a la presencia de cepas mas virulentas. Ante la ausencia frecuente de signos clinicos, el diagnostico de la toxoplasmosis congenita se basa en pruebas biologicas efectuadas en laboratorios especializados. En un nino cuya madre sufrio una seroconversion durante el embarazo, solo la desaparicion completa de las inmunoglobulinas G (IgG), anticuerpos antitoxoplasmicos, permite descartar el diagnostico de toxoplasmosis congenita. La conducta actual consiste en tratar al recien nacido durante el primer ano de vida con una asociacion de sulfadiazina-pirimetamina o de sulfadoxina-pirimetamina. Este tratamiento no esta exento de toxicidad y su eficacia no ha sido demostrada. Las lesiones oculares pueden aparecer o recidivar bajo tratamiento. Es fundamental, por tanto, prevenir la toxoplasmosis congenita, sobre todo ofreciendo consejos de higiene a las mujeres embarazadas seronegativas.

Infection during pregnancy is common and the developing fetal brain is vulnerable to vertical transmission due to immaturity of the fetal immune system. Infection is a major cause of multiple organ abnormalities, including the neuraxis, due to the neurotropism of the infectious agents. This review sets out to give an overview of fetal infection, review the general principles of the nature and timing of the infectious insult with respect to outcomes, review the neuroimaging of infection by ultrasound and magnetic resonance imaging (MRI), and review the various pathogens involved, including the two most common, cytomegalovirus (CMV) and Toxoplasma, and also other common viral and nonviral infections.

Resume Introduction Parallelement a la decroissance du nombre d’adoptions internationales en France, les enfants adoptes sont de plus en plus âges et parfois en mauvaise sante. Face a ce phenomene, nous assistons depuis quelques annees a une augmentation des demandes de consultations de specialistes avant l’adoption. Cette etude analyse les raisons de telles consultations. Methode Etude descriptive multicentrique prospective allant du 1 er janvier au 31 decembre 2013. Resultats Dix centres ont participe a l’etude, pour un total de 196 consultations pre-adoption. Dans 88 % des cas, le motif principal etait un avis medical sur dossier pour un enfant identifie (32 %) ou un avis sur les particularites sanitaires d’un pays, que celui-ci soit identifie (34 %) ou non (23 %). Dans 6 % des cas, le motif etait surtout socio-familial et pour les 5 % restant, l’obtention d’informations sur l’adoption en general et ses procedures. Dans plus de 40 % des cas, que ce soit l’enfant ou le pays qui soit identifie, la Russie etait l’objet de la consultation du fait de dossiers complexes et tres charges et du syndrome d’alcoolisation fœtale redoute et souvent non evoque. Conclusion La degradation de la sante des enfants adoptes est une preoccupation de plus pour les futurs postulants a l’adoption et il convient d’avoir une grande experience de l’adoption avant de se lancer dans ce type de consultation pour informer au mieux les postulants sans choisir a leur place.

The BMI1 polycomb protein regulates self-renewal, proliferation and survival of cancer-initiating cells essentially through epigenetic repression of the CDKN2A tumor suppressor locus. We demonstrate here for the first time that BMI1 also prevents autophagy in chronic myeloid leukemia (CML) cell lines, to support their proliferation and clonogenic activity. Using chromatin immunoprecipitation, we identified CCNG2/cyclin G2 (CCNG2) as a direct BMI1 target. BMI1 downregulation in CD34+ CML cells by PTC-209 pharmacological treatment or shBMI1 transduction triggered CCNG2 expression and decreased clonogenic activity. Also, ectopic expression of CCNG2 in CD34+ CML cells strongly decreased their clonogenicity. CCNG2 was shown to act by disrupting the phosphatase 2A complex, which activates a PKCζ-AMPK-JNK-ERK pathway that engages autophagy. We observed that BMI1 and CCNG2 levels evolved inversely during the progression of CML towards an acute deadly phase, and therefore hypothesized that BMI1 could support acute transformation of CML through the silencing of a CCNG2-mediated tumor-suppressive autophagy response.

La toxoplasmose est une infection parasitaire cosmopolite interessant entre le tiers et le quart de la population mondiale. Les sources de contamination multiples (viande, eau, terre, legumes) expliquent l'acquisition souvent precoce de l'infection au cours de la vie. L'infection acquise est le plus souvent benigne, asymptomatique ou a l'origine de lymphadenopathies. La toxoplasmose congenitale acquise apres infection maternelle en cours de grossesse concerne environ 300 enfants chaque annee en France. A la naissance, plus de 80 % d'entre eux n'ont pas de signes cliniques, mais un quart va developper des retinochoroidites. Les formes graves de toxoplasmose congenitale, neuro-oculaires ou disseminees, sont rares en France. Les atteintes oculaires, qu'elles soient acquises ou congenitales, sont plus severes et plus precoces dans d'autres pays, notamment en Amerique du Sud, en lien avec des souches plus virulentes. En l'absence frequente de signes cliniques, le diagnostic de la toxoplasmose congenitale repose sur des examens biologiques faits dans des laboratoires specialises. Chez un enfant ne d'une mere ayant fait une seroconversion en cours de grossesse, seule la disparition complete des immunoglobulines G (IgG), anticorps antitoxoplasmiques, permet de recuser le diagnostic de toxoplasmose congenitale. L'attitude actuelle consiste a traiter le nouveau-ne pendant toute la premiere annee de sa vie avec une association de sulfadiazine-pyrimethamine ou de sulfadoxine-pyrimethamine. Ce traitement n'est pas denue de toxicite et son efficacite n'a pas ete demontree. Des lesions oculaires peuvent apparaitre ou recidiver sous traitement. La prevention de la toxoplasmose congenitale basee sur des conseils d'hygiene a delivrer aux femmes enceintes seronegatives reste donc essentielle.

Resume Introduction En France metropolitaine, les leishmanioses cutanees s'observent le plus souvent chez des patients ayant sejourne au Maghreb ou en Amerique du Sud. Il est toutefois signale des leishmanioses cutanees dues aLeishmania infantum chez des patients apparemment non immunodeprimes qui n'ont jamais quitte la metropole. Observation Une fillette de huit ans, originaire de Haute-Savoie et n'ayant jamais sejourne outre-mer, presentait une ulceration crouteuse de la region sous-orbitaire droite evoluant depuis un an et resistant a divers traitements locaux. Les prelevements de la lesion confirmaient la presence de L. infantum. Sous traitement associant une cryotherapie a des injections intralesionnelles d'antimoniate de meglumine, la guerison etait obtenue en un mois. L'interrogatoire revelait des sejours reguliers dans les Pyrenees-Orientales, zone d'endemie du parasite. Discussion Devant une ulceration chronique sur une region decouverte, resistant aux traitements locaux chez un patient n'ayant jamais quitte la metropole, une leishmaniose cutanee doit etre evoquee et un sejour dans le Sud de la France, zone de transmission de L. infantum, doit etre recherche. En plus de l'examen direct du produit de curetage de la lesion, et de l'examen anatomopathologique, le diagnostic peut etre oriente par l'immuno-empreinte (Western-blot) et confirme par la PCR realisee sur un papier buvard prealablement applique sur la lesion. ________________________________________ Summary Background In France, cutaneous leishmaniasis is frequently seen in patients returning from North Africa or South America. Autochthonous leishmaniasis due to Leishmania infantum causes rather visceral forms. Nevertheless, cutaneous leishmaniasis caused by this parasite is occasionally seen in immunocompetent patients who have never been outside France. Patients and methods An 8-year-old girl living in the Haute-Savoie region and who had never travelled overseas presented with chronic ulceration of the right cheekbone that failed to regress under topical therapy. Laboratory tests demonstrated the presence of L. infantum. Following cryotherapy and intralesional injections of meglumine antimonite, the lesion resolved within a month. The patient's medical history revealed repeated journeys to the Pyrenees-Orientales region of southern France. Discussion For chronic ulceration on an uncovered area that does not resolve with topical therapy, cutaneous leishmaniasis should be considered in the differential diagnosis even if the patient has never left France. Trips to the South of France (an endemic region) should be sought in the history. In addition to direct examination of the product from curettage of the lesion and histopathology, non-invasive methods such as Western blotting with PCR run on filter paper impressions allow accurate diagnosis. Mots cles " Leishmania infantum; " Leishmaniose cutanee; " France; " Ulceration chronique; " PCR Keywords " Leishmania infantum; " Cutaneous leishmaniasis; " France; " Chronic ulceration; " PCR ________________________________________ Figures and tables from this article: Figure 1. Aspect de la lesion apres ablation de la croute. Figure options Figure 2. Guerison apres traitement.

In pediatric units, most of the intravenous medications are prepared by the attending nurse at the bedside that can be affected by an error margin, so can be imprecise. Despite the possible consequences of imprecise medications administration, published studies on the topic are scarce. The main objective of this study was to measure the difference between the prescribed vancomycine concentration and the actual concentration measured in the medication administered to the patient. The secondary objective was to determine which step in the preparation was linked to the difference in concentrations. It was a prospective study, setting in a pediatric and neonatal university hospital intensive care unit. Over a 3-month period, an aliquot from every preparation for continuous infusion of vancomycin, made at the bedside by a nurse, was collected and the modalities of the preparation noted. Vancomycin concentration was measured by high performance liquid chromatography. Sixty-four preparations, accounting for 24 patients (gestationnal age: 67 ± 75 weeks, weigh: 4.8 ± 6.5 kg) were included. Vancomycin concentrations ranged from 3.33 to 60.0mg/mL. Measured concentration were in mean 7% smaller than prescribed concentration (P

Apoptosis is mediated by cysteine-dependent, aspartate-directed proteases of the caspase family that proteolyse strategic intracellular substrates to induce cell suicide. We describe here that engagement of apoptotic processes by Fas triggering or by staurosporine stimulation leads to the caspase-dependent inactivation of the nuclear factor kappa B (NF-kappaB) pathway after cleavage of IKK1 (IkappaB kinase 1) and NEMO (NF-kappaB essential modulator), which are needed to transduce NF-kappaB activation signals. In this study, we have analyzed in more detail, the role of NEMO cleavage, as NEMO, but not IKK1, is important for the pro-survival actions of NF-kappaB. We demonstrate that NEMO is cleaved after Asp355 to remove the last 64 C-terminal amino acids. This short form was unable to rescue NF-kappaB activation by tumor necrosis factor-alpha (TNF-alpha) when transfected in NEMO-deficient cells. Consequently, inactivation of NEMO resulted in an inhibition of the expression of antiapoptotic NF-kappaB-target genes coding for caspase inhibitors (cIAP-1, cIAP-2) or adaptors of the TNF receptor family. NEMO-deficient Jurkat cells transiently expressing a non-cleavable mutant of NEMO were less sensitive to TNF-alpha-induced apoptosis. Therefore, downmodulation of NF-kappaB activation via the proteolytic cleavage of NEMO could represent an amplification loop for apoptosis.

Acute myeloid leukemia (AML) cells carry molecular defects that promote their leukemic proliferation, resistance to apoptosis and defect in differentiation. Pharmacological targeting of the nuclear factor kappaB (NF-kappaB) pathway has been shown to promote apoptosis of primary AML cells and to sensitize blasts to neoplastic drugs (Frelin, Blood 2005, 105, 804). The Fms-like tyrosine kinase 3 (FLT3), which sustains proliferation of normal hematopoietic progenitors is frequently overexpressed or mutated in AML patients. Using Ba/F3 murine pre-B cells transfected with various mutants of FLT3 (ITD, D835V, D835Y) and the MV4-11 human AML line, we show that normal or oncogenic stimulation of FLT3 led to activation of NF-kappaB. Pharmacological inhibition of either FLT3 with AG1296 or NF-kappaB with the small molecule inhibitor of IkappaB kinase-2 AS602868 reduced viability and triggered cell death. Moreover, AS602868 was also found to interfere directly with FLT3 kinase activation. AS602868 thus appears to target two different kinases that play a crucial role in the pathogenesis of AML, making it particularly attractive as a new therapeutical approach for AML.

Resume Nous rapportons une transmission intrafamiliale de Trichophyton violaceum au niveau cutane chez les parents d’une fillette presentant des lesions paucisymptomatiques au niveau du cuir chevelu. Observation La fillette de 11 mois, originaire d’Ethiopie, presente des lesions squameuses du cuir chevelu dues a T. violaceum , traitees successivement par ketoconazole shampoing, puis griseofulvine 125 mg/j pendant huit semaines. Parallelement, les parents de l’enfant presentent des lesions d’epidermatophyties circinees egalement attribuees a T. violaceum pour lesquelles ils recoivent un traitement local par isoconazole. Un traitement preventif par ketoconazole shampoing leur est egalement prescrit. L’examen clinique, ainsi que l’ecouvillonnage systematique du cuir chevelu d’enfants originaires de zones d’endemie, permettrait de reduire le risque de transmission intrafamiliale et au niveau de la collectivite de dermatophytes anthropophiles, en permettant l’utilisation precoce de mesures prophylactiques adaptees.

Dans cette etude nous avons etudie l'efficacite de la molecule antiretrovirale 2',3'-dideoxyinosine (Videx®) contre les kystes cerebraux dans un modele murin de toxoplasmose cerebrale due a une souche kystogene sauvage de Toxoplasma gondii. L'importance de l'excipient a ete egalement etudie Trois doses ont ete utilisees: 50, 100 et 150 mg/kg de poids corporel/jour ont ete preparees. Pour les doses 50 et 150 mg/kg, la poudre pure de 2',3'-dideoxyinosine a ete mise en suspension dans du Maalox® avant de l'administrer aux souris; la dose de 100 mg/kg/j a ete preparee en broyant des comprimes de Videx® et mise en suspension dans de l'eau Une diminution du nombre des kystes ainsi qu'une modification morphologique de ceux-ci ont ete observees des la plus faible dose. La plus importante diminution a ete observee pour la dose de 100 mg/kg/j. Au bout de 30 jours de traitement a cette dose, 65 % des kystes ont ete detruits par rapport au groupe temoin. Pour les doses de 50 et 150 mg/kg/j preparees avec du Maalox®, respectivement 36 % et 51 % des kystes ont ete detruits. La ddl exerce donc un effet sur les kystes cerebraux meme a faible dose. La formulation galenique influence son action puisque les doses preparees avec le Maalox® ont ete moins efficaces que celles preparees avec les comprimes broyes.

The effect of artesunate and its metabolite dihydroartemisinin against the cerebral cysts of Toxoplasma gondii was studied. In vitro experiments were performed with the THP-1 cell line and showed an inhibition of parasite growth of approximately 70% with 0.1-0.5 microg/ml of dihydroartemisinin for 96 hr. However, with artesunate, dihydroartemisinin, or a combination (50:50) of them, the number of tachyzoites decreased approximately 40-50% and they appeared motionless. Fifty-eight to 72 hr after washing of the tachyzoites and THP-1 cells in culture, parasitized cells reappeared. In vivo, the 50:50 artesunate-dihydroartemisinin combination produced a decrease in cerebral cysts of approximately 40% after only 5 days of treatment. However, transplantations into naive mice using brains of treated mice gave positive results.

Du fait de la frequence des voyages et des resistances de P. falciparum aux antipaludiques, l'acces palustre est devenu une pathologie frequente en metropole. En 1997, 2 798 cas de paludisme ont ete rapportes au Centre national de reference pour les maladies d'importation (CNRMI). La majorite de ces acces etaient dus a P. falciparum, espece faisant courir un risque mortel surtout en cas de diagnostic retarde. Le biologiste joue un role capital, car la clinique de l'acces palustre n'etant pas specifique, le diagnostic est essentiellement parasitologique. Pour les laboratoires polyvalents, rarement confrontes a ces demandes d'examens auxquelles ils doivent repondre en urgence, le frottis mince reste l'element cle du diagnostic. Apres avoir passe en revue les differentes techniques, nous proposons un algorithme diagnostique qui permettra au biologiste de rendre un resultat fiable en un minimum de temps.

We analyzed by in situ hybridization the spatiotemporal expression of dm-20, myelin basic protein (MBP) and 2'-3' cyclic nucleotide phosphodiesterase (CNP) during embryonic and postnatal development of the normal mouse and two plp/dm-20 mutants: the jimpy mouse and a transgenic mouse overexpressing the plp gene. In the central nervous system (CNS) of the normal mouse, dm-20 mRNA was detected at embryonic day (E)9.5 in the laterobasal plate of the diencephalon. The pattern of expression of CNP transcript was superimposable on that of dm-20, but appeared slightly later, at E12.5. MBP mRNA was detected even later (E14.5), and, in addition, only in the caudal (rhombencephalon and spinal cord) territories of expression of dm-20 and CNP. These observations support our previous proposals: (1) dm-20-expressing cells in the germinative neuroepithelium are precursors of oligodendrocytes, and (2) oligodendrocytes emerge from distinct pools of precursors along the neural tube (Timsit et al., 1995). In the jimpy mutant, despite the mutation in the plp gene, cells of the oligodendrocyte lineage developed normally. It is only at the time of myelin deposition that oligodendrocytes die. During embryonic development of the transgenic mutant overexpressing plp, there were no alterations in the spatiotemporal pattern or the level of expression of dm-20 in the CNS, in contrast to the higher levels of dm-20 observed in the peripheral nervous system (PNS).

Halofantrine is an antimalarial drug which is widely prescribed for the treatment of infections with chloroquine-resistant strains of Plasmodium falciparum. Chemically, it is a phenanthrene methanol, belonging to the aryl-amino-alcohol family. It has recently been recognized that this drug may induce rare but serious, cardiotoxic effects, including lengthening of the QTc interval, 'torsade de pointes' and induction of late ventricular potentials. These events are thought to be related to a quinidine-like action of the drug. In addition, severe haemolytic accidents have been reported, mimicking blackwater fever and indicating an immunological process. As a result of these side-effects, new guidelines for prescription and more cautious use of halofantrine, particularly as a stand-by treatment for febrile access among travellers, are required.

A rapid-flow cytofluorometric susceptibility test for in vitro amphotericin B testing of yeasts was evaluated and compared to the National Committee for Clinical Laboratory Standards (NCCLS) M27-T reference broth macrodilution method. The flow cytofluorometric method is based on the detection of decreased green fluorescence intensity of cells stained with DiOC5(3), a membrane potential-sensitive cationic dye, after drug treatment. Testing was performed on 134 clinical isolates (Candida spp. and Torulopsis glabrata). From the dose-response curve obtained for each isolate, three endpoints were calculated by computer analysis (the concentrations at which the fluorescence intensity was reduced by 50, 80, and 90%, i.e., 50% inhibitory concentration [IC50], IC80, and IC90, respectively). A regression analysis correlating these endpoints with the M27-T MICs showed that the best agreement was obtained with IC80. The flow cytofluorometric method showed good reproducibility with control strains. These initial results suggest that the flow cytofluorometric method is a valid alternative to the NCCLS reference method.

This study was designed to evaluate drug use and drug costs of treatment of 1112 AIDS patients at the Infectious Diseases Unit at F. Houphouet Boigny Hospital in Marseilles, France, between January 1, 1990 and December 31, 1994. All drug expenditures directly or indirectly related to AIDS treatment were recorded for both inpatients and outpatients. There were 1952 hospital stays. For each stay baseline characteristics including age, sex risk factors, costs, and duration of hospitalization were noted. Patients were mainly young male drug addicts around thirty years of age. Reason for admission was also noted. The overall number of admissions per year has decreased since 1991 probably due to development of outpatient care. The number of stays per patient per year has decreased since 1993 because of the use of more appropriate therapeutic and prophylactic protocols. The number of drugs used was high increasing from 750 in 1990 to 868 in 1994. Cost of treatment doubled between 1990 and 1994 due to the introduction of many expensive new drugs. Closer analysis showed that the greatest increase in expenditure involved 'antibiotic/antiviral', 'psychiatry/neurology' and 'specialized therapy'. Although not frequently prescribed, costly drugs such as immunoglobulins, hematopoietic growth factors, and parenteral nutrition solutions accounted for a high proportion of total costs. Since AZT, ddI and ddC were used mainly for outpatient treatment, their cost was low in inpatients. Cytomegalovirus-related retinitis, tuberculosis, and multiple infections were cost-intensive complications. The increasing number of cytomegalovirus infections underlines the need for cost evaluation and surveillance of this complication. This study demonstrates that cost of treating AIDS patients is rising due to the use of more and costlier drugs. This finding underlines the need to evaluate and compare new therapeutic modalities in terms of cost effectiveness.

The role of blood platelets in the pathogenesis of malaria remains unclear. In this study we investigated the role of experimentally caused thrombocytopaenia in chronic (Balb/C) and acute (CBA/J) Plαsmodium-berghei-induced murine malaria. A group of 30 Balb/C mice were rendered thrombocyto-paenic by injection of anti-mouse platelet serum given every 2 days from day 2 to day 8 after malaria infection. Compared with the control group, thrombocytopaenic mice showed a greater mortality. The Kaplan-Meier estimate of the risk of death was 4.37-fold higher in the thrombocytopaenic group. Parasitaemia and weight loss was in agreement with the protective effect of platelets. In the acute malaria model, the survival rate was less significant but the estimated risk of death was 1.7-fold higher in the treated group. These results suggesting a protective role of platelets in murine malaria are not in line with previous reports of the literature. Taken together our data suggest, however, that platelets, similarly to some inflammatory cytokines like TNF, play a dual role in the pathogenesis of malaria. Depending on experimental conditions, e.g. the time of onset of thrombocytopaenia, the beneficial role of platelets may outweigh the deleterious one.

The stability of amphotericin B 5 mg/mL in 5% dextrose ophthalmic solution prepared by the Hospital Pharmacy Service was studied in different conditions of storage and use. Admixtures of amphotericin B were aseptically prepared in low-density polyethylene dropper bottles. The stability of amphotericin B was evaluated in ophthalmic dropper bottles stored in a refrigerator, at room temperature, protected from, or exposed to, light. To simulate the effect of exposure to air, some ophthalmic dropper bottles were opened twice daily and two drops were removed. Immediately after preparation, samples were collected to determine the initial drug concentration by high-performance liquid chromatography and to assess pH, osmolality and sterility. The same tests were conducted after four, eight and 15 days of storage in ophthalmic containers opened daily and unopened after eight, 15, 30, 60, 75 and 120 days of storage. Samples were visually inspected daily for signs of physical incompatibility. An additional study was conducted in four ophthalmic containers collected in the ophthalmology unit after eight or 15 days of current patient use testing the same parameters. Ophthamlic containers stored in the refrigerator (the closed and the opened daily set) showed no loss or deterioration of amphotericin B during the corresponding period of storage (120 and 15 days, respectively). We observed precipitation and degradation after 13 days of storage in ophthalmic containers exposed to normal lighting conditions at room temperature, and after 16 days in ophthalmic containers protected from light. There was no appreciable change in pH or osmolality in any of the samples. Microbiological invesigation disclosed negative culture results for all samples. This study shows that aseptically prepared amphotericin B ophthalmic solution packaged in low-density polyethylene bottles can be stored safely for up to 120 days when unopened and stored at 4 deg C and protected from light, for 16 days when stored at 22 deg C and protected from light and for 13 days when stored at 22 deg C and exposed to light.

In France, cutaneous leishmaniasis is frequently seen in patients returning from North Africa or South America. Autochthonous leishmaniasis due to Leishmania infantum causes rather visceral forms. Nevertheless, cutaneous leishmaniasis caused by this parasite is occasionally seen in immunocompetent patients who have never been outside France.An 8-year-old girl living in the Haute-Savoie region and who had never travelled overseas presented with chronic ulceration of the right cheekbone that failed to regress under topical therapy. Laboratory tests demonstrated the presence of L. infantum. Following cryotherapy and intralesional injections of meglumine antimonite, the lesion resolved within a month. The patient's medical history revealed repeated journeys to the Pyrénées-Orientales region of southern France.For chronic ulceration on an uncovered area that does not resolve with topical therapy, cutaneous leishmaniasis should be considered in the differential diagnosis even if the patient has never left France. Trips to the South of France (an endemic region) should be sought in the history. In addition to direct examination of the product from curettage of the lesion and histopathology, non-invasive methods such as Western blotting with PCR run on filter paper impressions allow accurate diagnosis.

Myelination in the central nervous system requires synthesis by oligodendrocytes of enormous amounts of lipids and proteins for incorporation in the developing myelin membranes. To approach the regulatory events coordinating the transcriptional activation of the genes that encode myelin proteins, we examined control of the myelin basic protein (MBP) locus. MBP plays a major role in myelin compaction. During development, MBP is already expressed in mature non-myelinating oligodendrocytes. Here we show that, in transgenic animals in which the E. coli lacZ reporter gene is under the control of increasingly large portions (256, 1900 and 3200 bp) of the MBP promoter, 5’ of the initiation of transcription site, reporter gene expression was initiated after myelin formation had started. This delayed expression of the transgene compared to MBP, strongly suggests that premyelinating expression is dependent on regulatory elements located outside of the 3200 bp sequence studied, while expression occurring at the time of myelin formation is dependent on the proximal promoter sequence.

Ocular lesions of congenital toxoplasmosis may occur and relapse unpredictably even a long time after birth. There is no consensus concerning the necessity or timing of ophthalmologic follow-up for these patients. We surveyed adults with congenital toxoplasmosis followed regularly since birth, in order to learn their perceptions of this follow-up. The goal of this study was to provide doctors with patient-reported information on how they perceived the long-term monitoring of their disease.Enrolled patients were given a two-question questionnaire addressing the way they perceived the long-term follow-up and their attitudes toward continuing it. Eligible patients had to be 18 years or older and to have undergone ophthalmologic follow-ups, including funduscopy, every year since birth. The last ophthalmologic examination had to be within one year of the patient's inclusion in the study.Of the 102 patients finally included in the study, 98% stated that the follow-up was useful and 92% reassuring. Among the 11% of patients who found the follow-ups frightening, the proportion of patients with low visual acuity and low score on the visual function test was significantly higher than among the others. All patients except two wished to continue with regular follow-up.Without general agreement or guidelines on how patients with congenital toxoplasmosis should be monitored, the patient's wishes are important in making a decision. Our study brought out a clear fact; the majority of patients found long-term follow-up useful and reassuring and wished to continue.

CD43 (leukosialin, gpL115, sialophorin) is a major sialoglycoprotein widely expressed on hematopoietic cells that is defective in the congenital immunodeficiency Wiskott-Aldrich syndrome. It is thought to play an important role in cell-cell interactions and to be a costimulatory molecule for T lymphocyte activation. Using a metabolic 35SO4(2-) radiolabeling assay or biotinylation of cell surface proteins, we describe here that CD43 are sulfated molecules the glycosylation of which is altered in human immunodeficiency virus type 1 (HIV-1)-infected leukemic T cells of the CEM line. Hyposialylation of O-glycans and changed substitution on N-acetylgalactosamine residues are observed. The glycosylation defect is associated with an impairment of CD43-mediated homotypic aggregation which can be restored by resialylation. The hyposialylation of CD43 on HIV-1+ cells may explain the high prevalence of autoantibodies directed against nonsialylated CD43 that have been detected in HIV-1-infected individuals. A defect in glycosylation of important molecules such as CD43 or, as we recently described, CD45 may explain alterations of T cell functions and viability in HIV-1-infected individuals. In addition, a possible implication of hyposialylation in the HIV-1-infected cells entrapment in lymph nodes could be envisioned.

To determine virulence factors of isolates of Plasmodium falciparum and the potential role of cytokines in cerebral malaria, 46 Malagasy patients presenting with cerebral (n = 10), severe (n = 10), and uncomplicated (n = 26) malaria were enrolled in a study. The capacity of 21 of 46 P. falciparum isolates to form rosettes in vitro and to adhere to human umbilical vein endothelial cells (HUVECs) that express intercellular adhesion molecule-1 receptors and to C32 amelanotic melanoma cells that express mainly CD36 receptors was investigated together with the effects of tumor necrosis factor alpha (TNF-alpha), granulocyte macrophage-colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-6 alone and in two-by-two combinations on the cytoadherence of infected erythrocytes to HUVECs. Plasma levels of these cytokines were also measured in the patients at admission. The percentage of rosette formation was higher for the isolates from patients with cerebral (n = 6; 19.5%) and severe (n = 6; 30.5%) malaria than for those from patients with uncomplicated malaria (n = 9; 5%) (P < 0.002). The cytoadherence properties of the isolates did not differ among the three groups whatever the target cell used, but adherence to melanoma cells was systematically higher than that to HUVECs. Adhesion to HUVECs was increased more after TNF-alpha stimulation than after GM-CSF, IL-3, or IL-6 stimulation (P < 0.01). Only the combination of TNF-alpha and IL-3 enhanced cytoadherence more than TNF-alpha used alone (P < 0.02). No difference in the modulation of cytoadherence by cytokines was found in relation to the severity of the disease. TNF-alpha and IL-6 levels in peripheral blood were higher in the patients with cerebral and severe malaria than in the patients with uncomplicated malaria (P < 0.005). Most of the patients' sera contained little or no IL-3 or GM-CSF. Our results challenge the role of intercellular adhesion molecule-1 as the principal receptor mediating the cytoadherence of P. falciparum-infected erythrocytes and contrast with data obtained in the murine model.

In pediatric units, most of the intravenous medications are prepared by the attending nurse at the bedside that can be affected by an error margin, so can be imprecise. Despite the possible consequences of imprecise medications administration, published studies on the topic are scarce. The main objective of this study was to measure the difference between the prescribed vancomycine concentration and the actual concentration measured in the medication administered to the patient. The secondary objective was to determine which step in the preparation was linked to the difference in concentrations. It was a prospective study, setting in a pediatric and neonatal university hospital intensive care unit. Over a 3-month period, an aliquot from every preparation for continuous infusion of vancomycin, made at the bedside by a nurse, was collected and the modalities of the preparation noted. Vancomycin concentration was measured by high performance liquid chromatography. Sixty-four preparations, accounting for 24 patients (gestationnal age: 67 ± 75 weeks, weigh: 4.8 ± 6.5 kg) were included. Vancomycin concentrations ranged from 3.33 to 60.0mg/mL. Measured concentration were in mean 7% smaller than prescribed concentration (P10(-3)), with a large confidence interval (75.8%-120.4% of the prescribed concentration). Imprecision the preparations was much higher than this admitted for manufactured preparation. We could not highlight any factor related to the difference in concentrations, but one third of the preparation did not respect all the ISO 7886 standards for syringes use. Bedside vancomycin preparations, like preparations for other molecules, are far more imprecise than industrial intravenous medications. Our results urge that all pediatric intravenous medications should be made only by manufacturers or pharmacists. However, it also urged clinical studies, in parallel to pharmacodynamic and pharmacokinetic studies, to make intravenous treatments as accurate as they should be.

Despite the publication of guidelines for handling antineoplastic agents, measurable amounts of these drugs are still found at various hospital sites. In this context, the French cancer network ONCORA supported the present study to assess the impact of environmental contamination controls on the quality of practices during the preparation of cytotoxic drugs. The first part of the study was conducted at five voluntary hospitals. A total of 65 wipe samples of objects and surfaces were taken in the drug preparation rooms and analyzed for the presence of 5-fluorouracil (5-FU). Measurable amounts of 5-FU were detected in 21 samples (32%). Many surfaces within Biological Safety Cabinets and isolators were found contaminated (36%). The worse results were obtained on gloves and on the outside of infusion bags. The same method was applied during the second part of the study, conducted six months after the end of the first audit. Global contamination was reduced to 17%. This study shows that appropriate handling helps decrease the number of samples contaminated, making it possible to recommend these controls for evaluating and improving the quality of practices. Since 2007, the network's laboratory has extended its activities to all French hospitals interested in this quality assurance programme.

A longitudinal study was carried out in Burkina Faso to investigate the natural development of the immune response to Plasmodium falciparum malaria. Three bleedings were carried out before, during, and after the seasonal peak of transmission. Detailed antigen mapping and antibody prevalence of the 248 collected serum samples were established by immunoblotting on the basis of several epidemiological and biological parameters. An improved Western immunoblotting system was used to analyze up to 67 serum samples on each nitrocellulose sheet. This system allowed us to perform the entire study with strictly comparable conditions. Two different blood-stage antigens (exoantigens and somatic antigens) were used to analyze the distribution of different classes and subclasses of immunoglobulins according to the age of the individuals, the presence or absence of a malarial attack, the transmission period, the origin of parasite isolates, and the response to intraerythrocytic stages. Although this analysis emphasizes strong individual variations, reactions with two major antigens of 115 and 103 kDa were especially noted. These antigens induced high antibody levels and prevalences but were probably not involved in protection. The prevalence of immunoglobulin G (IgG) antibodies differed by isotype. Most of antigens stimulating IgG production were also responsible for the IgM antibody response. The role played by these antibodies in the development of natural immunity against malaria is discussed.

To date, only 20–30% of acute myeloid leukemia (AML) patients escape relapse to achieve long-term survival,1 demonstrating the crucial need for new targeted therapies.

Prescribing chemoprophylaxis against malaria is a medical act which is often controversial. The choice of appropriate molecule depends on destination, length of stay and tolerance to a given product. This latter aspect is difficult to evaluate. Among the three most commonly used drugs for malarial prevention--Savarine, Lariam and doxycycline--the first enjoys the reputation of being well tolerated, the second is perceived as being poorly tolerated and the third, little used until now, is suspected of serious complications. It is impossible to say by reviewing the available literature, which of these three molecules is indeed the least well tolerated, but the studies have followed heterogeneous evaluation methods. We propose that the various scientific associations concerned by the issue draw together an information sheet on side effects so as to encourage practioners to investigate all cases of intolerance of antimalarial drugs.

Congenital toxoplasmosis (CT) is the most frequently encountered congenital infection. The aim of the present review is to report about the long-term outcome of functional and morphological manifestations after an early confirmation of the diagnosis.We report on a cohort of patients with serologically confirmed CT, born between 1988 and 2001, who were followed up prospectively at a single centre (Institute of Parasitology, Hôpital de la Croix Rousse, University of Lyon). All patients underwent regular ophthalmological, parasitological and paediatric controls on a half-yearly or annual basis. Ocular manifestations were documented and visual acuity was assessed as far as was possible.1,506 seroconversions were diagnosed during pregnancy, and 327 of the live-born offspring were seropositive for CT. At the end of the study period, ocular manifestations occurred in 79 of the children (24 %), and in 7 cases (9 %) they were discovered during the first month of life. In 72 of the patients (91 %), the ocular manifestations developed between 1 and 151 months (median follow-up time: 6.3 +/- 3.7 years; range: 0.5 - 14 years) under medical treatment for the first year of life. At the end of the study period, information regarding visual acuity was available for 66 patients. In 55 of these (83 %), visual acuity was normal in both eyes, and in 11 (17 %), it was reduced below 0.5. In 1 child, both eyes were functionally affected, one severely.CT may not become manifest ocularly until after a decade, which underlines the necessity for long-term ophthalmological monitoring of the infected individuals. In our patients, the prognostic outcome after therapy for the first year of life was better than that reported in the literature. This favourable outcome may serve as a basis for patient counselling.

NF-kappaB (Nuclear Factor-kappaB) was described for the first time in 1986 as a nuclear protein binding to the kappa immunoglobulin-light chain enhancer. Since then, NF-kappaB has emerged as an ubiquitous factor involved in the regulation of numerous important processes as diverse as immune and inflammatory responses, apoptosis and cell proliferation. These last two properties explain the implication of NF-kappaB in the tumorigenic process as well as the promise of a targeted therapeutic intervention. This review focuses on the current knowledge on NF-kappaB regulation and discusses the therapeutic potential of targeting NF-kappaB in cancer in particular during radiotherapy.

Plasmodium falciparum drug resistance is a major problem in malaria endemic areas. Molecular markers and in vitro tests have been developed to study and monitor drug resistance. However, none used alone, can provide sufficient data concerning the level of drug resistance and to issue precise guideline for drug use policies in endemic areas. We propose real-time PCR for the simultaneous detection of pfcrt and pfmdr1 genes mutations. The aim of this study was not to provide definitive data concerning the rate of mutations in an endemic area, but to describe a powerful method allowing the detection of major pfmdr1 and pfcrt mutations.

Congenital toxoplasmosis (CT) may lead to serious neurological or sensory consequences. A serological screening of women at risk of acquiring toxoplasmosis became mandatory in France, first during the visit before wedding (1978), then during the visit for pregnancy declaration (1985) and at last with a monthly follow-up during pregnancy since 1992. The efficacy and the profitability of the program was never assessed, in spite of the modification of the epidemiological context. However medico-economical studies were conducted in countries in which no prevention program for CT was available to determine the interest of an antenatal screening similar to the French one or of other prevention strategies.Eight studies comparing at least two strategies were selected. Methodologies used in those studies were analyzed by two independent readers with the help of a standardized scale. A score was calculated for each study.Each study analyzed suffered from methodological limitations, in particular concerning the estimation of antenatal treatment efficacy, which could lead to invalidate their conclusion. The most reliable studies in regard to methodological guidelines, that is with the higher score, concluded that antenatal screening was not contributive. However, they could not be transposed directly in the present French situation, because of the difference of the epidemiological and economical context.Given the difficulty to obtain a clear conclusion, it seems necessary to perform a rigorous decision analysis to identify the more effective and acceptable program in terms of human and financial costs for preventing congenital toxoplasmosis.

To assess the consequences for the fetus of maternal toxoplasma infection acquired during the first 8 weeks of gestation and to set guidelines on how to manage these maternal infections.Data were prospectively analyzed on 360 pregnancies followed-up in our department due to a toxoplasma infection during the 8 first weeks of pregnancy. Estimates of the risk of fetal infection were based on all cases, including those which could not be followed up until infection was ruled out or confirmed. Severity of infection was estimated based on ultrasound findings during pregnancy, neonatal and long-term postnatal clinical, neurological and ophthalmologic work up.Out of the 360 included women, 336 gave birth to a live born child: 7 (2%) were infected, 302 (90%) were free of infection and follow-up was insufficient to conclude about the 27 (8%) remaining infants. The estimated risk of fetal infection ranged between 2 and 10% based on live born children and between 3 and 14% when the 24 interrupted pregnancies were included. At their last clinical evaluation at 70 months of age, all 7 children, including the 2 who had inactive peripheral eye lesions and the one who had a unique intracranial calcification were free of any ophthalmologic or neurological impairment.Our study confirms that in the event of a maternal infection during the first 8 weeks of pregnancy the risk of fetal infection is low and results mainly in a spontaneous termination of pregnancy. Future parents should be assured that conversely to a common opinion, the prognosis of congenital toxoplasmosis in live-born children is good. For these early maternal infections as for those acquired later, we recommend immediate treatment with spiramycin, monthly ultrasound surveillance, amniocentesis and treatment with pyrimethamine and sulphamides if the PCR is positive. Abortion should be restricted to cases with ultrasound lesions

Monthly serological screening of non immune pregnant women, required by law in France, is sometimes interrupted before delivery. As a consequence, late maternal infections resulting in a high risk of vertical transmission with potentially severe late sequelae are not detected.

The transcription factor nuclear factor (NF) kappaB is involved in the regulation of cell survival. NFkappaB is activated in many malignant tumors and seems to play a role in the resistance to cytostatic treatments and escape from apoptosis. We have studied the effects on NFkappaB activation of two topoisomerase poisons and DNA damaging agents that are used in chemotherapy: SN38 (7-ethyl-10-hydroxycamptothecin), the active metabolite of CPT11, and doxorubicin. In HeLa cells, both drugs activate NFkappaB using a preexisting pathway that requires a functional IkappaB-specific kinase complex, IkappaB-specific kinase activation, IkappaB-alpha phosphorylation, and degradation. Blocking NFkappaB activation by stable expression of a mutant super-repressor IkappaB-alpha molecule sensitized HeLa cells to the apoptotic actions of drugs and tumor necrosis factor. RNase protection assay analysis demonstrate that NFkappaB is involved in the regulation of a complex pattern of gene activation and repression during the cellular response of HeLa cells to topoisomerase poisons. The blockade of NF-kappaB activation seems to shift the death/survival balance toward apoptosis.