Etude de la prise en charge de l'infection à Helicobacter Pylori dans les villes de Pointe-Noire et de Brazzaville en 2015

Introduction : L'infection à Helicobacter Pylori affecte environ 50% de la population mondiale. Sa prévalence est plus élevée dans les pays en développement. Elle est à l'origine de pathologies gastroduodénales et son éradication est de ce fait recommandée.Nous avons réalisé une étude dont l'objectif a été d'évaluer les possibilités diagnostiques, thérapeutiques et la séroprévalence de l'infection à Helicobacter pylori dans les villes de Pointe-Noire et Brazzaville.Méthodes: Etude transversale descriptive et analytique menée de mars à septembre 2015.Résultats : Sur (7) sept tests existants et validés dans le monde; seuls (4) quatre tests étaient disponibles. La non disponibilité du test respiratoire à l'urée marquée au carbone 13 était à l'origine du non contrôle de l'éradication après traitement. Les protocoles d'éradication étaient la quadrithérapie séquentielle ou continue sur 10 jours sans sel de bismuth, non disponible au Congo-Brazzaville.Au total 130 patients ont été inclus; 54(41,5%) hommes et 76 (58,5%) femmes dont 121 (93,1%) patients étaient testés positifs à l'helicobacter pylori, avec une prédominance féminine. Le reflux gastro-oesophagien était la pathologie la plus représentée chez les patients testés positifs mais sans différence significative (P=0,287).Conclusion: L'insuffisance des tests diagnostiques de l'infection à Helicobacter pylori notamment du test respiratoire à l'urée marquée au carbone 13 au Congo-Brazzaville ne permettait pas aux praticiens de contrôler l'éradication. La quadrithérapie bismuthée n'était non plus disponible alors que la séroprévalence hospitalière reste élevée. Ainsi nous faisons le plaidoyer pour l'acquisition du test respiratoire à l'urée marquée au carbone 13 et la mise sur le marché des sels de Bismuth au Congo-Brazzaville

Evolution fatale à moyen terme d'une cirrhose virale C traitée par bithérapie pégylée

La prévalence de l'hépatite C au Congo-Brazzaville est élevée avec prédominance du génotype 4. Nous rapportons un premier cas clinique traité dont la réponse virologique en fin de traitement a été négative du fait que la patiente présentait tous les facteurs prédictifs de mauvaise réponse. L'évolution s'est faite vers les complications classiques mortelles. Les limites d'accès aux explorations et au traitement ont influencé négativement l'évolution. Nous venons de présenter un premier cas d'hépatite C traité à Pointe-Noire avec échec thérapeutique. Ce premier cas de notre expérience nous a permis de vivre la réalité de l'histoire naturelle et des difficultés de la prise en charge de l'hépatite C à Pointe-Noire

Modulation of human chondrocyte metabolism by recombinant human interferon.

OBJECTIVES: Interferon gamma (IFN gamma) is found to be elevated in the synovial fluid of patients with rheumatoid arthritis and osteoarthritis, suggesting its implication in joint disease pathogenesis. In this study, we investigated the effects of IFN gamma on the production of cytokines (IL-6, IL-8, IL-10), prostaglandin E(2)(PGE(2)), proteoglycans (PG), nitric oxide (NO), interleukin-1 receptor antagonist (IL-1ra) and stromelysin by non-stimulated and IL-1 beta-treated human chondrocytes. The role played by NO in the responses of chondrocytes to IFN gamma was also examined by incubation of chondrocytes with N(G)-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of NO synthase. METHODS: Enzymatically isolated human chondrocytes were cultured for 48 h in the absence or presence of IL-1 beta, IFN gamma or N(G)-monomethyl-L-arginine (L-NMMA) added solely or in combination. The productions of IL-6, IL-8, IL-10, IL-1ra and stromelysin were measured by enzyme amplified sensitivity immunoassays (EASIA). PG and PGE(2)were quantified by specific radioimmunoassays (RIA). Nitrite concentrations in the culture supernatants were determined by a spectrophotometric method based upon the Griess reaction. RESULTS: As expected, IL-1 beta highly stimulated NO, IL-6, IL-8, IL-10, IL-1ra, PGE(2)and stromelysin synthesis, but dramatically decreased PG production. NO, IL-6, IL-1ra and PGE(2)production by non-stimulated chondrocytes was dose-dependently increased by IFN gamma while PG production was inhibited. In the absence of IL-1 beta, IL-10 was undetectable in the culture supernatants. At the doses of 10 and 100 U/ml, IFN gamma markedly inhibited the constitutive and IL-1 beta-stimulated IL-8, IL-10 and stromelysin productions. Interestingly, IFN gamma synergized with IL-1 beta to increase NO, IL-6, IL-1ra and to depress PG production. As previously reported, the inhibition of NO synthesis by the competitive inhibitor L-NMMA led to enhancement of IL-6, IL-8 and PGE(2)production by IL-1 beta treated chondrocytes, but did not significantly modify IL-10, PG and MMP-3 productions. Inhibition of NO synthase significantly inhibited the stimulating effect of IFN gamma on IL-6 and IL-1ra but did not affect the inhibitory effect of IFN gamma on IL-8, PG or stromelysin production. CONCLUSIONS: These findings suggest that IFN gamma and IL-1 synergistically stimulate the production of IL-6, IL-1ra, NO and PGE(2)and inhibit PG synthesis. By contrast, IL-1 beta and IFN gamma have opposite effects on IL-8, IL-10 and stromelysin productions. These effects are not reversed by L-NMMA, suggesting that NO is not the principal mediator involved in responses of chondrocytes to IFN gamma.

Effects of nimesulide and sodium diclofenac on interleukin-6, interleukin-8, proteoglycans and prostaglandin E2 production by human articular chondrocytes in vitro.

OBJECTIVES: The aim of this study was to investigate the effects of two nonsteroidal anti-inflammatory drugs (NSAIDs), nimesulide and sodium diclofenac, on the production of proteoglycans (PG), prostaglandin E2 (PGE2) and cytokines (IL-6 and IL-8) by human articular chondrocytes in vitro. METHODS: Enzymatically isolated chondrocytes were cultured under constant agitation in a well defined culture medium. Specific radioimmunoassays were used to quantify PG and PGE2 production. Cytokine production (IL-6 and IL-8) was assayed by enzyme amplified sensitivity immunoassays (EASIAs). RESULTS: At a concentration of 3 micrograms/ml, nimesulide did not affect the PG production by chondrocytes. This concentration was superior to the highest level of nimesulide found in the synovial fluid of patients with rheumatoid arthritis 3 hours after the last oral administration of nimesulide (100 mg twice daily for 7 days). At 6 micrograms/ml a significant reduction in the PG content was obtained in the cellular phase in 5 out of the 8 cultures investigated. No similar effect was observed in the culture supernatants. Above this concentration nimesulide inhibited PG production in a dose-dependent manner. At concentrations ranging from 0.005 to 1 microgram/ml diclofenac did not significantly alter PG production. At therapeutic concentrations PGE2 production was totally inhibited by nimesulide, thus suggesting that PG inhibition is not linked to PGE2 production. Nimesulide inhibited PGE2 production by unstimulated (IC50 = 6 ng/ml) and IL-1 beta-stimulated (IC50 = 6.9 ng/ml) chondrocytes. At these concentrations, PGE2 production was fully inhibited by diclofenac. Furthermore, both nimesulide and diclofenac at therapeutic concentrations significantly decreased spontaneous and IL-1 beta-stimulated IL-6 production by human chondrocytes, but did not modify IL-8 production. CONCLUSION: From the results of this study we conclude that nimesulide and diclofenac at therapeutic concentrations are potent inhibitors of PGE2 and IL-6 production while they do not modify proteoglycan or IL-8 production.

Nitric oxide downregulates interleukin 1beta (IL-1beta) stimulated IL-6, IL-8, and prostaglandin E2 production by human chondrocytes.

OBJECTIVE: To investigate the effects of endogenously produced nitric oxide (NO) on interleukin 6 (IL-6), IL-8, prostaglandin E2 (PGE2), and proteoglycan production by human chondrocytes. METHODS: Human articular chondrocytes were isolated from their extracellular matrix by triple successive enzymatic digestion of the cartilage and cultured 48 h in a well defined culture medium. IL-6 and IL-8 were directly assayed into culture media by specific enzyme amplified sensitivity immunoassays. Proteoglycans and PGE2 were quantified by specific radioimmunoassays. Cell culture media were assayed for NO2 using a spectrophotometric assay based upon the Griess reaction. RESULTS: Unstimulated chondrocytes produced low levels of NO, IL-6, IL-8, and PGE2. Production was significantly stimulated by IL-1beta and lipopolysaccharide (LPS). As well, proteoglycan synthesis was profoundly inhibited by IL-1beta and LPS. Inhibition of NO synthesis with the competitive inhibitor NG-monomethyl-L-arginine (L-NMMA) led to enhancement of IL-6, IL-8, and PGE2 production stimulated by either IL-1beta alone or in combination with LPS, whereas the inhibition of proteoglycan production by IL-1beta was not modified by L-NMMA. CONCLUSION: LPS and IL-1beta stimulated IL-6, IL-8, and PGE2 production are downregulated by endogenously produced NO, which could limit the inflammatory reaction occurring in arthritis.

Immunoreactive somatostatin in the rat ovary.

Immunoreactive SRIH was detected in the rat ovary (15.6 pg/mg wet weight, 520 pg/mg protein) and was localized to the granulosa cells (168 +/- 6 pg/10(6) cells). Serial dilution studies showed parallelism of the inhibition curve for synthetic SRIH-14 and those of extracts of whole ovary and media conditioned by granulosa cells. The quantity of immunoreactive SRIH released into granulosa cell conditioned media decreased with time, while the intracellular content remained relatively constant. Gel chromatography showed peaks of immunoreactivity co-eluting with SRIH-14 (38%), SRIH-28 (31%) and a high molecular weight component. The addition of synthetic SRIH-14 stimulated meiotic maturation in cumulus-enclosed rat oocytes, with dose dependency being observed at SRIH-14 concentrations between 1 and 1000 pmol/l. No evidence of pre-pro-SRIH gene expression could be demonstrated in either rat ovary or testis using both Northern analysis and reverse transcriptase/polymerase chain reaction amplification of polyadenylated RNA. SRIH may be produced in the ovary during a specific stage of ontogeny or by an alternative gene. It is also possible that SRIH is actively taken up and stored by granulosa cells without being produced locally.

Studies on processing, particle formation, and immunogenicity of the HIV-1 gag gene product: a possible component of a HIV vaccine.

Antigens in a particulate conformation were shown to be highly immunogenic in mammals. For this reason, the particle forming capacity of derivatives of the HIV-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. Moreover, p55 gag was expressed in combination with the protease (p55-PR) or with the entire polymerase (p55-pol), respectively. Budding of 100-160 nm p55 core particles, resembling immature HIV-virions, was observed in the eucaryotic expression systems only. In comparison to the vaccinia virus driven expression of p55 in mammalian cells, considerably higher yields of particulate core antigen were obtained by infection of Spodoptera frugiperda (Sf9) insect cells with the recombinant Autographa californica nuclear polyhedrosis (AcMNPV) baculovirus. Mutation of the NH2-terminal myristylation signal sequence prevented budding of the immature core particles. Expression of the HIV p55-PR gene construct by recombinant baculovirus resulted in complete processing of the p55 gag precursor molecule in this system. The introduction of an artificial frameshift near the natural frameshift site resulted in constitutive expression of the viral protease and complete processing of p55, both in Escherichia coli and in vaccinia virus infected cells. Interestingly, significant processing of p55 resembling that of HIV infected H9 cells could also be achieved in the vaccinia system by fusing the entire pol gene to the gag gene. Moreover, processing was not found to be dependent on amino-terminal myristylation of the gag procursor molecule, which is in contrast to observations with type C and type D retrovirus. However, complete processing of p55 into p24, p17, p9 and p6 abolished particle formation. Purified immature HIV-virus like particles were highly immunogenic in rabbits, leading to a strong humoral immune response after immunization. Empty immature p55 gag particles represent a noninfectious and attractive candidate for a basic vaccine component.

Truncated versions of the two major Epstein-Barr viral glycoproteins (gp250/350) are secreted by recombinant Chinese hamster ovary cells.

The expression of the two Epstein-Barr virus (EBV) major membrane proteins gp250/350 (MA-BLLF1) on the surface of recombinant CHO clones cannot be amplified by methotrexate (MTX) selection, perhaps due to toxic effects of these membrane proteins. After removal of sequences encoding the part of the glycoproteins responsible for membrane anchorage, the gp250/350 is secreted into the medium. Following selection with MTX, this construct allows the amplification of the expression products. Besides the possible use of these proteins in protection experiments, they can also be used as antigens for diagnosis, which opens an efficient approach for control of EBV-related neoplasias by early therapy.

Expression of the Epstein-Barr virus major membrane proteins in Chinese hamster ovary cells.

The gene of the major membrane antigen (gp250/350) of the Epstein-Barr virus (EBV) was isolated and inserted under the control of the SV40 early promoter into a eukaryotic expression vector, which allows selection for dhfr+ phenotype. Following transfection with this vector, Chinese hamster ovary cells express on their surfaces proteins immunologically similar to the major EBV membrane antigen. The transcript encoding gp250/350 is processed by partial splicing similarly but more efficiency than in B95-8 cells from which the DNA originates.

An immunoprecipitation blocking assay for the analysis of EBV induced antigens.

A technique for the analysis of EBV antigens in extracts of unlabelled EBV infected cells has been developed. Using this technique we have demonstrated that the EBV early antigen complex consists of several proteins and is not completely expressed in chemically induced Raji cells. Studies with a range of sera from patients with infectious mononucleosis and nasopharyngeal carcinoma have shown that despite similar titers by the indirect fluorescent antibody test different populations of proteins were precipitated by different sera.