In vitro models for neuropathic pain phenotypic screening in brain therapeutics.

The discovery of brain therapeutics faces a significant challenge due to the low translatability of preclinical results into clinical success. To address this gap, several efforts have been made to obtain more translatable neuronal models for phenotypic screening. These models allow the selection of active compounds without predetermined knowledge of drug targets. In this review, we present an overview of various existing models within the field, examining their strengths and limitations, particularly in the context of neuropathic pain research. We illustrate the usefulness of these models through a comparative review in three crucial areas: i) the development of novel phenotypic screening strategies specifically for neuropathic pain, ii) the validation of the models for both primary and secondary screening assays, and iii) the use of the models in target deconvolution processes.

Generation of an equine biobank to be used for Functional Annotation of Animal Genomes project.

The Functional Annotation of Animal Genomes (FAANG) project aims to identify genomic regulatory elements in both sexes across multiple stages of development in domesticated animals. This study represents the first stage of the FAANG project for the horse, Equus caballus. A biobank of 80 tissue samples, two cell lines and six body fluids was created from two adult Thoroughbred mares. Ante-mortem assessments included full physical examinations, lameness, ophthalmologic and neurologic evaluations. Complete blood counts and serum biochemistries were also performed. At necropsy, in addition to tissue samples, aliquots of serum, ethylenediaminetetraacetic acid (EDTA) plasma, heparinized plasma, cerebrospinal fluid, synovial fluid, urine and microbiome samples from all regions of the gastrointestinal and urogenital tracts were collected. Epidermal keratinocytes and dermal fibroblasts were cultured from skin samples. All tissues were grossly and histologically evaluated by a board-certified veterinary pathologist. The results of the clinical and pathological evaluations identified subclinical eosinophilic and lymphocytic infiltration throughout the length of the gastrointestinal tract as well as a mild clinical lameness in both animals. Each sample was cryo-preserved in multiple ways, and nuclei were extracted from selected tissues. These samples represent the first published systemically healthy equine-specific biobank with extensive clinical phenotyping ante- and post-mortem. The tissues in the biobank are intended for community-wide use in the functional annotation of the equine genome. The use of the biobank will improve the quality of the reference annotation and allow all equine researchers to elucidate unknown genomic and epigenomic causes of disease.

Acceptability of children's testing for HIV infection during routine care activities in the Pediatrics Department of Sourô Sanou Teaching Hospital in Bobo-Dioulasso (Burkina Faso).

BACKGROUND: In order to contribute to the fight against the pediatric HIV infection, we have assessed, through a study in which we have systematically proposed to carry out children's testing, the rate of acceptability and the feasibility of children's HIV testing during the routine activities of the department. We have also analyzed the reasons for the acceptability or the refusal of the child's HIV testing by the accompanying person. METHODS: The study took place from May to September 2015 including all the parents/legal guardians of any child aged 0 to 14 years coming for a consultation or who was hospitalized in the Pediatric Department of Souro Sanou Teaching Hospital. Counseling sessions conducted by community health workers focused on informing and proposing the principle of child testing. After obtaining the verbal and informed consent of the accompanying person, the first test was performed with Determine® by a hospital health worker. A second SD Bioline®/ImmunoCombII® test was performed if the first test was positive. With children aged less than 18 months, after a positive antibody test, we resorted to PCR for confirmation. RESULTS: A total of 848 accompanying persons, 568 of whom were female, underwent a pre-test interview during which the HIV test was offered to them. The mean age of accompanying persons was 30 (25.5 to 38) years; 747 accompanying persons (88.1%) accepted the testing of their child. We have found an influence of the accompanying person's religion (P=0.02) and the type of accompanying person on the acceptability of children's testing. Mothers were more willing to accept the test compared to other accompanying persons (P=0.002). The main reason for refusing the child's testing was the absence of one of the child's parents, mainly the father whose opinion was needed. The test was positive for HIV1 in 10 children. CONCLUSION: In health centers, getting the informed consent from parents to test their children is a big challenge. However, our study shows that this is possible, through the high rate of acceptability obtained.

[Missed opportunities for immunization in pediatric unit of the Bobo-Dioulasso hospital (Burkina Faso)]. / Les occasions manquees de vaccination en milieu hospitalier pediatrique de Bobo-Dioulasso (Burkina Faso).

AIM: This study was to determine the prevalence of missed opportunities for immunization in the pediatrics unit of the Souro Sanou University Hospital. METHODS: This was a cross-sectional descriptive study conducted from May to June 2010 involving children aged 0-11 months, accompanying persons and health personnel. Self-questionnaires were administered to health personnel in order to gage their knowledge of vaccination practices. RESULTS: 177 children including 101 girls and 49 health workers were recruited. The vaccination rate for children targeted by the Expanded Programme on Immunization (EPI) was 38% in the city of Bobo-Dioulasso and the rate of missed vaccination occasions was 50%. The low level of parental education, ignorance of EPI targeted diseases and a poor organization of the vaccination unit were noted. CONCLUSION: The overall strategy for vaccine administration should be rethought in a country with limited resources such as Burkina Faso to optimize immunization, EPI target diseases contributing to the mortality and morbidity in these regions.

BUT: L'objectif était de déterminer la fréquence des occasions manquées de vaccination dans le département de pédiatrie du Centre Hospitalier Universitaire Sourô Sanou de Bobo-Dioulasso. MÉTHODE: il s'agissait d'une étude transversale descriptive de mai à juin 2010 enrobant les enfants âgés de 0­11 mois, les personnes accompagnatrices, les agents de santé. Des auto-questionnaires ont été administrés au personnel de santé afin de récolter leur connaissance sur la pratique de la vaccination. RÉSULTATS: Avaient été inclus dans l'étude, 177 enfants dont 101 filles ainsi que leurs accompagnants et 49 agents de santé. Le taux de couverture vaccinale des enfants cibles du programme élargi de vaccination (PEV) dans la ville de Bobo-Dioulasso était de 38% et le taux des occasions manquées de vaccination était de 50%. L'enquête avait noté un faible niveau d'instruction des parents, la méconnaissance des maladies cibles du PEV ainsi qu'une mauvaise organisation de l'unité de vaccination. CONCLUSION: La stratégie globale d'administration des vaccins devra être repensée dans les pays à ressources limitées comme le Burkina Faso afin d'optimiser la vaccination, les maladies cibles du PEV contribuant à alourdir la mortalité et la morbidité dans ces contrées.

[Study of factors associated to therapeutic failure of children infected by HIV1 under antiretroviral treatment in the Sanou Sourô University Hospital of Bobo-Dioulasso (2007-2013)]. / Etude des facteurs associes a l'echec therapeutique au cours du suivi des enfants infecte par le VIH1 sous traitement antiretroviral au CHU-Sanou Souro de Bobo-Dioulasso (2007­2013).

OBJECTIVE: To determine the prevalence of therapeutic failure and the associated factors in children aged 6 months to 15 years old with HIV 1 and regularly visiting the Sanou Sourô University Hospital (SSUH) of Bobo-Dioulasso. METHODS: A retrospective study was conducted concerning the children followed in the SSUH between February, 2007 and February 2013 infected by HIV 1 under ARV for at least six months. The diagnosis of therapeutic failure was defined according to the WHO criteria. RESULTS: the population of study was 311 infected patients, 53.8 % were male. The average age was of 108 months ± 67, and 62.0 % had a rate CD4 higher than 200 cells/ µ L; 43.5 % of the patients were at class III / IV of the WHO classification. Prevalence of the failure was at 19.6 %. The female genital organ (RR: 0.49 IC95 %: [0.24-0.99] p = 0.03), respect for the treatment ≤ 95 % (RR 0.37 IC95 % [0.15-0.92] p=0.04), the death of the mother (RR: 0.33 IC95 % [0.09-1.21] p=0.04) and the class WHO advanced (III / IV) (RR 0.26 IC95 % [0.09-0.77] p=0.02) was associated to the therapeutic failure. CONCLUSION: prevalence of therapeutic failure was high for the children being followed at the Sanou-Sourô University Hospital of Bobo-Dioulasso. The female genital organ, respect for the treatment ≤ 95 %, death of the mother, and the advanced WHO classification stages (III / IV) were associated with therapeutic failure.

OBJECTIF: Déterminer la prévalence et les facteurs associés à l'échec thérapeutique chez les enfants âgés de six mois à 15 ans dépistés positifs au VIH-1 et suivis régulièrement au CHU-Sanou Sourô de Bobo-Dioulasso. MÉTHODES: Il s'agissait d'une étude de cohorte rétrospective ayant concerné les enfants suivis au CHUSS entre février 2007 et février 2013 infectés par le VIH-1 sous ARV depuis au moins six mois. Le diagnostic de l'échec thérapeutique a été défini selon les critères de L'OMS. RÉSULTATS: La population d'étude était de 311 patients infectés, 53,8% étaient de sexe masculin. L'âge moyen était de 108 mois ± 67, et 62,0% avait un taux CD4 supérieur à 200 cellules/µL ; 43,5% des patients étaient au stade III/IV de l'OMS. La prévalence de l'échec était de 19,6%. Le sexe féminin (RR : 0,49 IC95% : [0,24­0,99] p= 0,03), l'observance < 95% (RR :0,37 IC95% [ 0,15­0,92] p=0,04), le décès de la mère (RR : 0,33 IC95% [ 0,09­ 1,21] p=0,04) et le stade OMS avancé (III/IV ) (RR :0,26 IC95% [0,09­0,77] p=0,02) étaient associés à l'échec thérapeutique. CONCLUSION: la prévalence de l'échec thérapeutique était élevée chez les enfants suivis au CHU-Sanou-Sourô de Bobo-Dioulasso. Le sexe féminin, l'observance ≤ 95%, le décès de la mère, et le stade OMS avancé (III/IV) étaient associés à l'échec thérapeutique.

[Impact of vaccination on admissions for Haemophilus influenzae b meningitis from 2004 to 2008 in Bobo Dioulasso, Burkina Faso]. / Impact de la vaccination sur la méningite à H. influenzae b de 2004 à 2008, à Bobo Dioulasso (Burkina Faso).

OBJECTIVE: Vaccination against Haemophilus influenzae type b was introduced in Burkina Faso on 1st January 2006. This study thus sought to determine the impact of the first 30 months of vaccination on admissions for Hib meningitis in the department of pediatrics at the Sourô-Sanou University Hospital in Bobo Dioulasso. METHODS AND PATIENTS: Retrospective study of children aged zero to 14 years hospitalized from 1st January 2004 to 30th June 2008 for acute bacterial meningitis (laboratory-confirmed). RESULTS: During the study period, 416 children were admitted for acute bacterial meningitis. The bacterium isolated was identified in 386 cases and unidentified in 30 cases. Hib meningitis accounted for 42.3 % of the cases of identified bacterial meningitis before the introduction of the vaccine (2004 to 2005). This rate declined to 11.8 % for the first 30 months of vaccination (p < 0.001). No cases of Hib meningitis have been reported in the first half of 2008. CONCLUSION: Admissions for Hib meningitis in the Department of Pediatrics have practically disappeared two years after the introduction of the Hib vaccine into Burkina Faso's expanded program on immunization.

[Acute renal failure and Plasmodium falciparum malaria: a case report]. / Insuffisance rénale aiguë et paludisme à Plasmodium falciparum : à propos d'un cas.

Malaria is an endemic disease caused by one of the several Plasmodium species. Severe malaria is mainly due to Plasmodium falciparum in highly endemic areas. Acute renal failure (ARF) is a criterion of malaria severity as defined by WHO. Often observed in adults, particularly in India and Southeast Asia, this complication remains a rare complication of malaria in children. We report a case of oliguric ARF that occurred in a 7-year-old girl a few days after the onset of fever. The vascular obstruction by parasitized erythrocytes often causing tubular necrosis is the primary mechanism of renal failure. As a possible diagnosis, hemolytic uremic syndrome, renal failure and quartan hemoglobinuric nephropathy are other possible causes of renal failure in malaria. Renal biopsy, which was not performed in our patient, would have been a great help, but was not available. The outcome was favorable with recovery of renal function after 3 weeks of diuretic therapy. This development is not always the rule and the prognosis depends on early diagnosis and treatment options.

[Preoperative diet and complete dentures in Ivory Coast]. / Diététique préopératoire et prothèse adjointe totale en Côte d'Ivoire.

The toothless patients of developing country are a real problem of nutrition. The practitioners are in the habit of doing the prostheses restoration wit hoof thinking of alimentation. In this work, authors show the energetic and protein malnutrition of the toothless patients of Cote d'lvoire and propose a hyperprotein diet with their alimentation habit in order to prepare the psychic and physic site who must receive complete prostheses.

Differential usage of RNA templates by the rotavirus "in vitro" replication system.

Rotavirus genome synthesis is a two-step process; the synthesis of the eleven viral mRNAs and the production of the complementary strand (minus strand synthesis) yielding the double stranded genomic RNA (dsRNA). Purified rotavirus open cores are a helpful tool to study this replication event. These open cores can use viral dsRNA and mRNAs as template for RNA synthesis. In the present communication it was demonstrated that open cores prefers to use viral mRNA as template instead of the genomic dsRNA. The use by the open cores of both templates can be blocked through two independent strategies. In the presence of ssRNAs molecules, the use of the viral dsRNA template is completely blocked without affecting the minus strand synthesis. Nevertheless, an antisense oligonucleotide strategy previously reported to block the minus strand synthesis of the target gene do not affect the use of the dsRNA template. These results suggest the recognition of different signals in each template by the open core protein complex.

Translation enhancer in the 3'-untranslated region of rotavirus gene 6 mRNA promotes expression of the major capsid protein VP6.

The eleven rotavirus mRNAs contain 5'-cap structures and most end with the 3'-consensus sequence 5'-UGACC-3'. The UGACC functions as a common translation enhancer (3'-TE-con) that upregulates viral protein expression through a process mediated by the nonstructural protein NSP3. To address the possibility that gene-specific enhancers are also contained in the untranslated regions (UTRs) of the rotavirus mRNAs, we used rabbit reticulocyte lysates to investigate the translation efficiencies of analog RNAs containing viral-specific 5'-and 3'-UTRs and the open reading frame for chloramphenicol acetyltransferase. These experiments combined with the analysis of full-length viral RNAs and RNAs containing 3'-truncations showed that a highly active enhancer was present near the 5'-end of the 139-nucleotide 3'-UTR of the gene 6 mRNA (3'-TEg6). The 3'-TEg6 represents a functionally independent enhancer, as no other portion of the gene 6 mRNA was required for its activity. The 3'-TEg6 differs significantly from the 3'-TE-con in that the gene 6-specific enhancer does not require viral protein for activity and is formed by a sequence unique to only one of the eleven viral mRNAs. Together, our findings suggest that the 3'-UTR of the gene 6 mRNA contains two TEs, one is gene-specific (3'-TEg6) and the other is common to nearly all rotavirus genes (3'-TE-con). The activity of the 3'-TEg6 is likely important for directing the efficient translation of the gene 6 mRNA at levels sufficient to provide the 780 copies of VP6 necessary for the assembly of each progeny virion.

NSP5 phosphorylation regulates the fate of viral mRNA in rotavirus infected cells.

Elucidation of the function of the non-structural rotavirus proteins during infection is difficult in the absence of a reverse genetic system. To study the role of NSP5, nonstructural phosphoprotein NSP5, we constructed a reassortant strain (SACC11) in the SA11 background that harbours a heterologous segment 11 encoding a variant protein (h-NSP5). Cells infected by SACC11 produced viral polypeptides at earlier times than SA11 infected cells while showing less accumulation of genomic dsRNA. These changes suggested that NSP5 might direct viral messenger RNA to protein synthesis or genome replication. Distinct patterns of proteins were shown to form complexes with NSP5 in co-immunoprecipitation studies with SA11 and SACC11 infected cells. Recombinant h-NSP5 from either bacteria or eucaryotic cells migrated faster in PAGE suggesting that it was hypophosphorylated. Indeed, the kinase inhibitor H-7 enhanced translation of viral proteins in SA11 but not SACC11 infected cells suggesting that NSP5 function in the regulation of the fate of viral positive strand RNA is mediated by phosphorylation.

Identification of adenovirus 7h heterogeneity in the E3 region.

Adenovirus genotype 7h was previously reported to be originated from a recombination event between adenovirus genotypes 7p and 3p. Based on those findings, further characterization of other adenovirus 7h strains become important to determine whether all adenovirus 7h strains arose from a single recombinational event. To explore such a possibility, 160 clinical isolates were studied after developing a PCR assay using a primer set designed to amplify the region corresponding to E3-7,7 Kd of adenovirus ADV 7p and E3-9 Kd of adenovirus 3p. The assay was able to differentiate most of the subgenus B strains from adeno 7h with the genotype 3d. The study of several adenovirus 7h clinical isolates revealed the existence of three variants of adeno 7h. One of the variants, 7h3, shows a high degree of similarity with gene E3-9 Kd of ADV 3p, but lacks the corresponding AUG codon. Our results suggest that more than one recombination event may explain the detection of three different types of adenovirus 7h. The genotype variants of adeno 7h were detected in different years, indicating that the recombination events took place independently from each other. The study of the recombination region may allow further understanding of the function of several viral polypeptides in the immune response, and understanding the mechanism involved in virulence associated to adenovirus 7h.

Identification of sequences in rotavirus mRNAs important for minus strand synthesis using antisense oligonucleotides.

The core of the rotavirion consists of three proteins, including the viral RNA polymerase, and 11 segments of double-stranded (ds)RNA. The RNA polymerase of disrupted (open) cores is able to catalyze the synthesis of dsRNA from exogenous viral mRNAs in vitro. In this study, we have identified sequences in exogenous viral mRNAs important for RNA replication using antisense oligonucleotides. The results showed that oligonucleotides complementary to the highly conserved 3'-terminal sequence of rotavirus mRNAs prevented all but basal levels of dsRNA synthesis. Notably, we observed that the addition of oligonucleotides which were complementary to nonconserved sequences present either at the 5'- or 3'-end of a viral mRNA effectively inhibited its replication without interfering with the replication of other viral mRNAs present in the same replication assay. Thus, the nonconserved sequences in rotavirus mRNAs contain gene-specific information that promotes RNA replication. The fact that antisense oligonucleotides inhibited dsRNA synthesis indicates that the strandedness (single- versus double-stranded) and secondary structure of the viral mRNA template are factors that affect the efficiency of minus strand synthesis.

[Use of a simple polymerase chain reaction combined with a heteroduplex mobility assay to characterize the non coding 5' region of hepatitis C virus]. / Uso de una reacción de polimerasa en cadena simple combinada con un ensayo de movilidad de heteroduplexes en la caracterización de la región 5' no codificante del virus de la hepatitis C.

BACKGROUND: One of the most used methods for the characterization of hepatitis C virus strains is the use of a nested polymerase chain reaction (PCR) with a restriction fragment length polymorphism (RFLP) assay. Sometimes, RFLP results do not differentiate new strains. There are other more complex methods and only the sequencing of the PCR fragment allows a correct characterization of the strain. AIM: To report the detection of hepatitis C virus using a single PCR assay of the 5' non codifying region. MATERIAL AND METHODS: Thirty five serum samples coming from patients with chronic hepatitis or blood donors were assayed for hepatitis C virus. RESULTS: The reported method increases the PCR sensitivity through the combination of polyacrylamide gel electrophoresis and silver staining of amplified products. This allowed the semi quantitative estimation of viral load and the characterization of amplified products through their electrophoretic motility. These PCR products were used in a heteroduplex motility assay; allowing the discrimination between sequences of different genotypes. CONCLUSIONS: Heteroduplex assays can be used to characterize the 5' non codifying region of the hepatitis C virus for routine laboratory purposes.

Structural domains of streptokinase involved in the interaction with plasminogen.

Two fragments of recombinant streptokinase, comprising amino acids Val143-Lys293 (17-kDa rSK) or Val143-Lys386 (26-kDa rSK), were cloned and expressed in Escherichia coli, purified to homogeneity and their interactions with plasmin(ogen) were evaluated. Both 17-kDa rSK and 26-kDa rSK bound to plasminogen with a 1:1 stoichiometry and with affinity constants of 3.0 x 10(8) M-1 and 12 x 10(8) M-1, respectively, as compared to 6.3 x 10(8) M-1 for the binding of intact recombinant streptokinase to plasminogen. Binding of 17-kDa rSK to plasminogen-Sepharose was displaced by addition of increasing concentrations of recombinant streptokinase, whereas bound recombinant streptokinase was not displayed by 17-kDa rSK. In equimolar mixtures of plasminogen and 26-kDa rSK, the appearance of amidolytic activity as monitored with a chromogenic substrate, was significantly delayed compared to the equimolar mixture with recombinant streptokinase (60% of the maximal activity after 30 min, compared to maximum activity within < or = 2 min). In contrast, no amidolytic activity was generated in equimolar mixtures of plasminogen and 17-kDa rSK. Plasminogen was rapidly activated by catalytic amounts (1:100 molar ratio) of recombinant streptokinase (60-70% within 10-15 min), whereas only 4% of the plasminogen was activated within 60 min with 26-kDa rSK, and no plasmin was generated with 17-kDa rSK. Complexes of plasmin with 17-kDa rSK or with 26-kDa rSK were very rapidly inhibited by alpha 2-antiplasmin (apparent second-order inhibition rate constant of approximately 2 x 10(7) M-1 s-1), whereas the complex with recombinant streptokinase was resistant to inhibition. With 26-kDa rSK, inhibition by alpha 2-antiplasmin resulted in dissociation of the complexes and recycling of functionally active 26-kDa rSK to other plasminogen molecules; 17-kDa rSK, in contrast, remained associated with the plasmin-alpha 2-antiplasmin complex. These findings suggest that different regions of the streptokinase molecule are involved in binding to plasminogen, in active-site exposure, and in impairment of the inhibition of plasmin by alpha 2-antiplasmin. Thus, the 17-kDa region spanning Val143-Lys293 in streptokinase mediates its binding to plasminogen but does not induce activation. Furthermore, this region does not interfere with the inhibition of the complex with plasmin by alpha 2-antiplasmin.

Kappa-opioid receptor mediated antinociception in rats is dependent on the functional state of dihydropyridine-sensitive calcium channels.

The modulatory effect of the dihydropyridine Ca2+ channel antagonist nimodipine on the analgesic action of the kappa-opioid receptor agonist U-69,593 was analyzed using the tail-flick test in rats. The antinociceptive effect of U-69,593 (0.25-4 mg/kg) was antagonized by L-type Ca2+ channel blockade with nimodipine (200 microgram/kg, i.p.), the ED50 being increased from 1.4 to 7.3 mg/kg. On the contrary, when an increase in the density of these channels was induced by means of chronic and simultaneous treatment with nimodipine (1 microgram/h, 7 days) and sufentanil (2 micrograms/h, 8 days), the analgesic effect of U-69,593 was potentiated by 5-fold. Our results suggest a functional coupling between kappa-opioid receptors and L-type Ca2+ channels in nociception.

[The immunogenic properties and prophylactic efficacy of a live polyvalent influenza vaccine in children 5 to 14 years old]. / Izuchenie immunogennykh svoistv i profilakticheskoi éffektivnosti zhivoi grippoznoi polivalentnoi vaktsiny u detei 5--14 let.

Production lots of a live influenza vaccine made of strains A/47/T (N1H1), A/47/6/2 (H3N2), and B/60/32 were used for vaccination of 3663 children aged from 5 to 14 years inoculated twice with monovaccines, a trivaccine made of the above strains, or placebo. Both mono- and polyvaccine were practically areactogenic. An average per cent of subjects with a significant rise in antibody titres to the respective three antigens was 60%. The efficacy of the vaccination was 31.0-42.8% for monopreparations and 36.3% for the trivaccine. The studies showed the possibility and expedience of using for children the live influenza vaccine in the form of a polyvalent preparation including current influenza type A and B viruses.

[The inoculation properties of live recombinant influenza vaccine types A and B used separately and jointly in children 3 to 14]. / Privivochnye svoistva zhivoi grippoznoi rekombinantnoi vaktsiny tipov A i B pri razdel'nom i sovmestnom primenenii detiam 3--14 let.

The reactogenicity and immunizing activity of vaccine influenza virus A (H1N1) and B strains used as mono- and bi-preparations in children of 3 to 14 years was studied. No increased reactogenicity after the use of bivaccine was observed in the children. Febrile reactions as well as 9 other clinical symptoms which could indicate the reactogenicity of the vaccines were identical for mono- and bivaccine and corresponded to the requirements of the technical documents for the vaccine. The optimal conditions for the evaluation of the immunogenicity of the B component by HI test were developed, and the necessity of using additionally the enzyme immunoassay for this purpose is substantiated. The above method demonstrated that the immunogenicity of the live influenza type A and B vaccine was high in children. No significant inhibition of immunological parameters was observed when the two viruses were combined in the bivaccine.