Onchocerciasis control in the Democratic Republic of Congo (DRC): challenges in a post-war environment.

OBJECTIVE: To evaluate onchocerciasis control activities in the Democratic Republic of Congo (DRC) in the first 12 years of community-directed treatment with ivermectin (CDTI). METHODS: Data from the National Programme for Onchocerciasis (NPO) provided by the National Onchocerciasis Task Force (NOTF) through the annual reports of the 21 CDTI projects for the years 2001-2012 were reviewed retrospectively. A hypothetical-inputs-process-outputs-outcomes table was constructed. RESULTS: Community-directed treatment with ivermectin expanded from 1968 communities in 2001 to 39 100 communities by 2012 while the number of community-directed distributors (CDD) and health workers (HW) multiplied. By 2012, there were ratios of 1 CDD per 262 persons and 1 HW per 2318 persons at risk. More than 80% of the funding came from the fiduciary funds of the African Programme for Onchocerciasis Control. The cost of treatment per person treated fell from US$ 1.1 in 2001 to US$ 0.1 in 2012. The therapeutic coverage increased from 2.7% (2001) to 74.2% (2012); the geographical coverage, from 4.7% (2001) to 93.9% (2012). Geographical coverage fell in 2005 due to deaths in loiasis co-endemic areas, and the therapeutic coverage fell in 2008 due to insecurity. CONCLUSIONS: Challenges to CDTI in DRC have been serious adverse reactions to ivermectin in loiasis co-endemic areas and political conflict. Targets for personnel or therapeutic and geographical coverages were not met. Longer term funding and renewed efforts are required to achieve control and elimination of onchocerciasis in DRC.

The African Programme for Onchocerciasis Control: impact on onchocercal skin disease.

OBJECTIVES: To assess the long-term impact of the African Programme for Onchocerciasis Control on itching and onchocercal skin disease (OSD). METHODS: Seven study sites in Cameroon, Sudan, Nigeria and Uganda participated. Two cross-sectional surveys were conducted of communities meso- and hyper-endemic for onchocerciasis before and after 5 or 6 years of community-directed treatment with ivermectin (CDTI). Individuals were asked about any general health symptoms including itching and underwent full cutaneous examinations. Onchocercal skin lesions were documented according to a standard classification. RESULTS: Five thousand one hundred and ninety three people were examined in phase I and 5,180 people in phase II. The presence of onchocercal nodules was a strongly significant (P < 0·001) risk factor for all forms of onchocercal skin disease: APOD (OR 1·66); CPOD (OR 2·84); LOD (OR 2·68); reactive skin lesions (OR 2·38) and depigmentation (OR 3·36). The effect of community-directed treatment with ivermectin was profound. At phase II, there were significant (P < 0·001) reductions in the odds of itching (OR 0·32), APOD (OR 0·28); CPOD (OR 0·34); reactive skin lesions (OR 0·33); depigmentation (OR 0·31) and nodules (OR 0·37). Reduction in the odds of LOD was also significant (OR 0.54, P < 0.03). CONCLUSIONS: This first multi-country report of the long-term impact of CDTI reveals a substantial reduction in itching and OSD. APOC operations are having a major effect in improving skin health in poor rural populations in Africa.

[African programme for onchocerciasis control (APOC): entomological aspects of Onchocerca volvulus transmission by Simulium neavei in the Basin of Sankuru (Eastern Kasai, Democratic Republic of Congo)]. / Aspects entomologiques de la transmission d'Onchocerca volvulus par Simulium neavei dans le bassin du Sankuru (Kasaï oriental, République Démocratique du Congo).

As part of a study to evaluate the long-term impact of community-directed treatment with ivermectin (CDTI), baseline entomological data on the transmission of onchocerciasis in the forest zone of the Eastern Kasai Province were collected from July 1998 to January 1999. Species of the Simulium neavei complex were the only vectors found in the site during the study. Nuisance activity was low with a mean biting rate of only 32.5 bites/man/day. The mean parturity rate was 29.4%, the mean rate of infected females was 14.5% and the mean number of infective Onchocerca larvae per 1000 parous flies was 659. These findings clearly show that the S. neavei population at the study site has high vector capability and constitutes a large microfilarial reservoir. The entomological features found in the study area are consistent with a hyper-endemic zone.

[Epicardial atrial pacing using an active fixation bipolar endocardial lead in children].

Good performance was observed over 10 years after implantation of bipolar epicardial atrial pacing using an active fixation bipolar endocardial lead in 3 pediatric patients with congenital heart block. The bipolar endocardial lead which was supposed to be fixed transvenously was implanted on the atrial surface by first screwing the lead's helix into the myocardium. The catheter was then laid down on the atrial surface, and both electrodes were wrapped by the atrial tissue. The good performance of this pacing lead seemed to depend on stable positioning of the electrode. This lead is superior to the commercially available, and steroid eluting epicardial bipolar pacing lead, on the point of its small size in head and body, and could be applied to dual chamber pacing in smaller children.

[African Program for Onchocerciasis Control (APOC): Onchocerca volvulus transmission by Simulium damnosum s. l. in two regions of the Central African Republic]. / Programme africain de lutte contre l'onchocercose (APOC): transmission d'Onchocerca volvulus par Simulium damnosum s. l. dans deux régions de la République Centrafricaine.

Entomological baseline data were collected in the villages of Zinga and Boali-Falls in Central African Republic (CAR) in view of the long term impact assessment of community-directed treatment with ivermectin (CDTI). Morphological determinations revealed that flies caught in both sites belong to the sub-group Sou/Sq. In Boali, the nuisance was relatively high with biting rates averaging 243 bites/man/day, with a parous rate of 61.6% and a crude annual transmission potential (ATP) of 8,259 infective larvae/man/year; and the average number of infective larvae per 1,000 parous flies was 177. In Zinga, the mean biting rate was 191 bites/man/day, with a parous rate of 51.6%, a crude ATP of 3,422, and 86 infective larvae per 1,000 porous flies. In conclusion, the vectorial capacity and the entomological indices recorded are characteristic of high onchocerciasis transmission zones. However, some of the infective larvae found, maybe of animal origin, need identification to better determine the real level of endemicity.

[African Programme for Onchocerdasis Control (APOC): Onchocerca Simulium squamosum in two regions in the Republic of Cameroon]. / Programme africain de lutte contre l'onchocercose (APOC): intensité de la transmission d'Onchocerca volvulus par Simulium squamosum dans deux régions de la République du Cameroun.

In 1998, the African Programme for Onchocerciasis Control (APOC) decided to launch a long-term impact assessment of its operations. This paper reports the baseline entomological data collected throughout a whole year in two sites of Cameroon (Kahn and Bolo). The Simulium populations of the two study sites were characterized by parous rates of 7.2% and 33.5% respectively and infectivity by O. volvulus of 31 and 190 infective larvae per 1000 parous flies respectively The Annual Transmission Potentials (ATP) were respectively 523 and 9972 infective larvae per man and per year in Kahn and in Bolo. The Simulium populations studied in both sites, even though the ATP in Kahn is 19 times lower than that of Bolo, showed a pattern of an onchocerciasis hyperendemic zone in terms of vector capacities and entomological indices.

[Onchocerciasis in the area of Lastourville, Gabon. Clinical and entomological aspects]. / L'onchocercose dans la région de Lastourville, Gabon. Aspects cliniques et entomologiques.

The African Programme for Onchocerciasis Control (APOC) has implemented a series of surveys aimed at evaluating the long-term impact of its activities. The region of Lastourville (Gabon) is one of the selected sites for this study. A total of 886 persons was examined for skin lesions, and 459 out of them participated in detailed ocular examinations. Blackflies were collected during one year and dissected. Although the focus was found to be hypoendemic (prevalence of nodules: 7.7%), the frequency of onchodermatitis was relatively high. The lesions of the anterior segment of the eye were rare, but the prevalence of optic nerve disease, and of choroido-retinal lesions reached 5.2 and 2.7%, respectively. The annual transmission potential (2,171 infective larvae per man) was high, when compared with the results recorded in the human population. This may be due to the presence of Onchocerca spp. of animal origin in the blackflies. These results indicate that in the area of Lastourville, though regarded as the main focus of onchocerciasis in Gabon, the disease is relatively mild.

Sequence analysis of the agaB gene encoding a new beta-agarase from Vibrio sp. strain JT0107.

An agarase gene (agaB) was cloned from genomic DNA of Vibrio sp. strain JT0107. Analysis of the 3200 nucleotide sequence just before the agarase 0107 gene (agaA) which existed in genomic DNA of Vibrio sp. strain JT0107 revealed a putative single open reading frame coding for 955 amino acids. Comparison of the deduced amino acid sequence of AgaB to that of agarase 0107 revealed the existence of partially highly homologous regions. A part of this gene was expressed in Escherichia coli to yield a protein with agarase activity. This is the first report of evidence by genetic analysis that at least two different kinds of agarases exist in strain JT0107.

Human 5-lipoxygenase associates with phosphatidylcholine liposomes and modulates LTA4 synthetase activity.

A Ca2+ and a phosphatidylcholine (PC) as stimulatory factors to human 5-lipoxygenase (5-LO) were assessed to examine aspects of the regulatory mechanism of 5-LO. In the presence of Ca2+ (1 microM or less), PC liposomes distinctly stimulated the dual activities of 5-LO for the production of 5-HPETE from arachidonate and for its subsequent conversion to LTA4. At the same concentration of Ca2+, 5-LO was found to bind to PC liposomes. As with 5-LO activities, the binding was dependent on the range of Ca2+ concentration. The conversion ratios of 5-HPETE to LTA4 were dependent on PC liposome concentration and reached a maximum of 50% conversion. Among the four cell membrane lipids examined, PC liposomes demonstrated the highest conversion ratio of 5-HPETE to LTA4 by 5-LO. Most of the arachidonate added to the reaction mixture localized in PC liposomes. These results confirm that the intracellular increase of Ca2+ concentration causes 5-LO to associate with the cell membrane and perform an interfacial reaction. They also suggest that this binding of 5-LO to the cell membrane enhances the subsequent conversion from 5-HPETE to LTA4.

A structural study of calcium-binding equine lysozyme by two-dimensional 1H-NMR.

Since 1H-NMR spectra of the calcium bound form (holo) and the calcium free form (apo) of equine lysozyme have an overall similarity, the folded structure of apo equine lysozyme seems to be similar to the holo structure at 25 degrees C and pH 7.0, even at low ionic strengths except for subtle conformational change. However, calcium titration experiments showed that a number of resonances change by a slow exchange process. The changes saturated at one calcium ion per one lysozyme molecule, and no more change was observed by further addition of calcium ions. This shows that just one calcium ion binds to equine lysozyme. To make assignments for these changed proton resonances, two-dimensional 1H-NMR studies, correlated spectroscopy (COSY), two-dimensional homonuclear Hartmann-Hahn spectroscopy (HOHAHA) and nuclear Overhauser effect spectroscopy (NOESY) were carried out. A structural model of equine lysozyme based on the crystal structure of human lysozyme was estimated and used to assign some resonances in the aromatic and beta-sheet regions. It was possible to use some proton signals as a probe to determine the specific conformational change induced by calcium ions. The calcium binding constant KCa was estimated from calcium titration experiments in which changes in the proton signal were monitored. The log KCa value was found to be on the order of 6-7, which is in agreement with the calcium binding constant determined by fluorescence probes. This means that the protons are affected by specific calcium binding.

Escherichia coli ribosome is inactivated by Mirabilis antiviral protein which cleaves the N-glycosidic bond at A2660 of 23 S ribosomal RNA.

Ribosome-inactivating proteins (RIPs) are known to inactivate eukaryotic ribosomes, which results in the inhibition of protein synthesis, but there has been no evidence that they inactivate the ribosomes of Escherichia coli. Recently, Mirabilis antiviral protein (MAP), a RIP, has been shown to inhibit the protein synthesis of E. coli as well as eukaryotes. To elucidate its mechanism, E. coli ribosomes treated with MAP were analyzed by polyacrylamide/agarose composite gel electrophoresis and RNA sequencing using reverse transcriptase with DNA primer. The 23 S rRNAs, with an A260 value for ribosomes of 15, were completely cleaved in vitro by a 30 minute treatment with MAP at a concentration of 100 nM at 37 degrees C and a subsequent treatment with aniline. However, they were not affected by ricin A-chain under the same conditions. The primer extension of DNA polymerization stopped before A2660 of 23 S rRNA in RNA sequencing. Furthermore, both 16 S and 23 S rRNAs were cleaved by the MAP and aniline treatments when naked E. coli rRNAs were used as substrates, and the primer extension stopped before bases A2660 and A1014, respectively, in RNA sequencing. As the A2660 region has been shown to interact with the elongation factors EF-Tu and EF-G these results indicate that MAP cleaves the N-glycosidic bond at A2660 in E. coli 23 S RNA resulting in the inactivation of the ribosome.

Crystallization and preliminary X-ray crystallographic analysis of Mirabilis antiviral protein.

Mirabilis antiviral protein is a single-chain ribosome-inactivating protein purified from the tuberous root of Mirabilis jalapa L. We obtained several forms of crystals of the protein by the hanging drop vapor diffusion method, but most of these crystals were not suitable for X-ray crystallography. After refining the growth conditions, crystals of crystallographic quality were grown in 20-microliters droplets of an equi-volume mixture of 1.5% (w/v) protein solution and a reservoir solution containing 49 to 50% (w/v) ammonium sulfate and 50 mM-ammonium citrate (pH 5.4) at room temperature. Addition of 2 mM-adenine sulfate reduced twinning and "crystal shower". The resulting trigonal crystals diffract beyond 2.5 A resolution using a rotating anode X-ray generator. The space group was determined to be P3(1)21 or P3(2)21 (a = b = 103.9.A, c = 134.6 A, alpha = beta = 90 degrees, gamma = 120 degrees) based on their precession photography of h0l and hk0 zones. There seems to be three monomers in an asymmetric unit for VM = 2.51 A3/Da.

Crystallization and preliminary X-ray crystallographic studies of recombinant human leukotriene A4 hydrolase complexed with bestatin.

Recombinant human leukotriene A4 hydrolase complexed with bestatin, an inhibitor of metalloprotease, has been crystallized by the hanging drop vapor diffusion method using 0.1 M phosphate buffer (pH 6.5) and 50 to 54% saturated ammonium sulfate. The orthorhombic crystals belong to the space group I222 or I2(1)2(1)2(1) with unit cell dimensions of a = 273.6 A, b = 261.3 A and c = 52.9 A. They diffract beyond 2.5 A resolution and a native data set up to 3 A resolution has been collected on an imaging plate Weissenberg camera using synchrotron radiation.

[Delayed onset malignant hyperthermia after a closure of ventricular septal defect].

An 18 month-old girl was diagnosed as ventricular septal defect (VSD) with mild aortic valve prolapse. She underwent a closure of VSD. Intra-and early postoperative course was uneventful. However, 20 hours after surgery, sudden bradycardia led to cardiac arrest and strong muscle rigidity was seen. Hyperkalemia and metabolic acidosis rapidly progressed and resuscitation was failed. Extracorporeal life support and continuous hemodialysis were initiated, but the patient died with multiple organ failure on 5th postoperative day. Her clinical course supported the diagnosis of delayed onset malignant hyperthermia. Histopathological findings of muscle biopsy were consistent with rhabdomyolysis, and immunopathological stains demonstrated changes as in a Duchenne type muscular dystrophy carrier. Delayed onset malignant hyperthermia is an extremely rare complication of general anesthesia. We should be aware of this lethal condition, which occurs with a certain time lag after surgery, especially when the patient has possible background of myopathy.

[Cor triatriatum diagnosed in adulthood; report of a case].

Cor triatriatum is a rare congenital cardiac anomaly especially in adulthood. A 68-year-old female was diagnosed as a cor triatriatum classified to Lucas-Schmidt IA, severe degree of mitral regurgitation and atrial fibrillation. Resection of the abnormal diaphragm in the left atrium and miral valve replacement were performed. Although the reason of sudden death of this patient after discharge is unknown, surgical intervention for atrial fibrillation should have performed to prevent a thromboembolism in such cases.

Expression of a cDNA encoding human 5-lipoxygenase under control of the STA1 promoter in Saccharomyces cerevisiae.

A yeast-expression vector utilizing the STA1 promoter was constructed, and shown to be useful for the expression of a heterologous gene. A cDNA, encoding human 5-lipoxygenase (5LO), was inserted into the vector and expressed in Saccharomyces cerevisiae. The enzyme (yh5LO) produced in the transformant was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A4 synthase activities, which were stimulated by Ca2+ and ATP. The N-terminal end of yh5LO contained five extra amino acids not present in 5LO purified from human leukocytes. A human 5LO-secretion vector containing the STA1 signal sequence was also constructed. When this hybrid gene was expressed in S. cerevisiae, its product was glycosylated and accumulated in the fractions related to the secretory pathway.

Expression of human 5-lipoxygenase cDNA in Escherichia coli.

A cDNA for human 5-lipoxygenase (5LO) was inserted into the vector pKC (constructed from pKK223-3 by replacing its replication origin with that of pUC18) and expressed in Escherichia coli. The enzyme expressed was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A4 synthase activities, which were stimulated by Ca2+ and ATP. Its molecular mass (78 kDa) and NH2-terminal sequence were identical with those of 5LO purified from human leukocytes. The availability of the expression system will facilitate further studies on its regulation and the reaction mechanism of the enzyme.

Endotoxin transduces Ca2+ signaling via platelet-activating factor receptor.

Lipopolysaccharide (LPS) is a pathogenic substance causing severe multiple organ failures and high mortality. Although several LPS binding proteins have been identified, the molecular mechanism underlying the LPS signaling pathway still remains obscure. We have found that the LPS-induced Ca2+ increase in platelets and platelet aggregation is blocked by selective platelet-activating factor (PAF) receptor antagonists, thus suggesting a cross-talk between LPS and the PAF receptor. Next, we confirmed this hypothesis using the cloned PAF receptors [(1991) Nature 349, 342-346; (1991) J. Biol. Chem. 266, 20400-20405] expressed in Xenopus oocytes and Chinese hamster ovary (CHO) cells. In both systems, cells responded to LPS only when PAF receptors were expressed, and specific PAF binding was successfully displaced and reversibly dissociated by LPS. PAF receptor activation by LPS may represent a novel important pathway in the pathogenesis of circulatory collapse and systemic thrombosis caused by endotoxin.

A radioreceptor binding assay for platelet-activating factor (PAF) using membranes from CHO cells expressing human PAF receptor.

A simple and reproducible radioreceptor assay (RRA) has been developed using membranes from CHO cells which can stably express human platelet-activating factor (PAF) receptor. The CHO cells expressing the PAF receptor, termed CHO.1F8, showed a significant intracellular Ca2+ response to PAF, and the same binding properties to [3H]WEB 2086, a PAF antagonist, as reported (Kd, 13.6 +/- 1.9 nM; Bmax, 2.5 +/- 0.4 pmol/mg protein (n = 6)). A competitive binding assay was done using the CHO.1F8 cell membranes and [3H]WEB 2086. The minimum detectable dose of PAF was 0.3 nM (approximately 30 pg per well) and the assay was highly specific for PAF. This method makes it possible to handle large numbers of samples rapidly and simultaneously, since the receptor membrane is prepared in advance and the binding assay can be completed within 3 h. Using this method, we have determined the production and cell association of PAF in human neutrophils.

Inhibitory effects of 3-nitro-2,4,6-trihydroxybenzamides on Epstein-Barr virus early antigen induction.

Inhibitory effects of a series of 3-nitro-2,4,6-trihydroxybenzamides on Epstein-Barr virus early antigen (EBV-EA) induction were examined using Raji cells. Some of the tested compounds showed highly inhibitory activity, the N-octyl amide derivative being the most active among them. These results suggest the possibility that 3-nitro-2,4,6-trihydroxybenzamides might be listed as novel inhibitors of tumor promotion.