Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ)最新文献

Genomic variations of herpes simplex viruses type 2 (HSV 2) isolated from Chiang Mai, Thailand and Okinawa, southernmost part of Japan were studied. The genomic polymorphism of 40 HSV 2 Chiang Mai strains and 10 HSV 2 Okinawan strains was analyzed by means of the variations of cleavage sites and electrophoretic mobilities of DNA fragments after digestion with 4 restriction endonucleases (RE (BamH I, Kpn I, EcoR I, and Bgl II)). Using the main 6 variable RE cleavage sites, HSV 2 strains were classified into 10 major groups. The strains categorized into group 1 and 9 were predominant in Chiang Mai. Groups 1, 3, 4, 5, 6 and 7 were found only in Chiang Mai, but contained only a few strains each. Groups 2 and 10 were found only in Okinawa, whereas groups 8 and 9 were found in both Chiang Mai and Okinawa.

We report the clinical and parasitological effects of a modified treatment regimen for suramin. Twenty adult males received up to 5 g (72.5 to 84.7 mg/kg) of suramin over 36 days. Detailed clinical and laboratory examinations were done before treatment and then at intervals over 2 years. Nodules were removed at 6, 13, 26 and 52 weeks for histology. Systemic tolerance was good. Anterior segment inflammation was however common and 2 patients required intervention to prevent posterior synechiae. No new posterior segment lesions developed; a rare improvement occurred in one patient with papillitis. Proteinuria, mostly mild, occurred in nearly all patients. Previously unreported renal glycosuria was documented in one patient. Microfilariae in the skin and anterior chamber did not change significantly for 5 or more weeks after which rapid reductions occurred. Ocular parasites were absent at 2 years and skin microfilariae were near zero. Peripheral blood eosinophil counts fell in parallel with those of microfilariae in the skin and anterior chamber and were normal at one and two years. These findings at 2 years may provide indirect evidence of a macrofilaricidal or a permanent chemosterilant effect on the adult worms. Nodule examination revealed an embryotoxic effect from week 6, a lethal effect on the male worms from month 3 and on the female worms from month 6 after treatment started. At one year 34% of the female worms examined were alive. Thus total doses of suramin in the range 72.5 to 84.7 mg/kg have only a modest lethal effect on the female worms. Suramin remains a restricted drug and a suitable replacement is urgently needed.

To aid the development of a macrofilaricidal agent for Onchocerca volvulus, the African bovine parasite, O. ochengi, was evaluated as a drug screen by testing three known filaricidal drugs. Groups of five Zebu cattle, naturally infected with more than 15 palpable O. ochengi nodules in the ventral skin, were treated with either suramin (10 mg/kg/day i.v. for 6 days), ivermectin (200 micrograms/kg, s.c.), CGP 20376 (20 mg/kg orally) or left untreated as controls and examined at intervals up to 137 days post-treatment (d.p.t.). After ivermectin treatment, microfilarial densities in the skin decreased within one week to virtually zero and remained at a very low level. A similar rapid and profound reduction was seen after CGP 20376 treatment, but by 137 d.p.t. microfilarial skin densities were approaching pre-treatment levels. With suramin, skin microfilarial densities fell to very low levels after 12 weeks but rose slightly by 137 d.p.t. Effects on the macrofilariae were assessed by sequential nodulectomies at -3 and 28, 84 and 137 d.p.t. By 137 d.p.t. embryogenesis was almost completely interrupted in the CGP 20376 and ivermectin treated animals, although not in the suramin treated group, but in all three groups the majority of remaining intrauterine microfilariae were pathologically altered. Degenerating intrauterine microfilariae accumulated in the ivermectin and in the CGP 20376, but not in the suramin treated worms. The motility of male and female worms was not reduced by any treatment except for female worms at 84 d.p.t. with CGP 20376. Viability of the worms as indicated by the MTT-formazan reduction assay was not reduced in any of the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Stilbamidinium hexachloroiridiate was found trypanocidal in vitro against Trypanosoma brucei brucei IPP at 600 microM after a 1 h incubation period and 30 microM after 24 h. This activity was confirmed in mice with a subcutaneous treatment at 20 mg/kg in a single dose. It was then evaluated on T.b. brucei murine CNS model. At the early stage, a subcutaneous treatment at 2 mg/kg/day x 5 cured 50% mice where-as one single dose at 10 mg/kg was completely inactive. Higher doses failed to cure the mice. Nevertheless, hexachloroiridiate salt of stilbamidine was 3.3 fold less toxic than dihydrochloride salt. Although the compound appeared inactive at the late stage of the murine trypanosomiasis, the difference of toxicity justified its evaluation on the early stage of sheep trypanosomiasis. The compound was trypanocidal at 2 mg/kg in a single dose when administered 8 days after infection. The study of iridium serum kinetic showed that stilbamidinium hexachloroiridiate was distributed rapidly according to a monocompartmental model. Moreover, iridium persisted in serum for a long time. The compound in aqueous suspension with 1% carboxymethylcellulose acted therefore as a controlled release system with a bioavailability allowing its trypanocidal action at the early stage.

A cell viability assay, using fluorescence measurements has been developed for the screening of new compounds against African trypanosomes. 2',7'-Bis-(carboxyethyl)-5(6)-carboxyfluorescein-pentaacetoxymethyles ter (BCECF-AM), an esterase substrate, was used in the assay as a marker for cell viability. Fluorescence was quantified using an automated fluorescence scanner for multi-well plates. Trypanosoma brucei rhodesiense, T. congolense, T. evansi and T. equiperdum from continuously growing cultures were exposed to various concentrations of trypanocidal drugs for an incubation period of 72 h at 37 degrees C. Then BCECF-AM was added to the cell suspensions and after 60 minutes the fluorescence of the trypanosome suspension was measured using the Millipore Cytofluor 2300 fluorescence scanner, at 485 nm excitation and 530 nm emission wavelengths. Results of kinetic studies of the hydrolysis of the non-fluorescent BCECF-AM in trypanosomes showed that BCECF-AM is readily cleaved by non-specific esterases to a highly fluorescent product. Drug concentrations causing 50% inhibition of fluorescence (IC50-values) were measured fluorimetrically. Minimum inhibitory concentration (MIC) was determined microscopically.

In 1993, a three arm double-blind controlled trial was implemented in French Polynesia, to compare the tolerance and efficacy of single doses of the combination ivermectin (IVR) 400 micrograms.kg-1 plus diethylcarbamazine (DEC) 6mg.kg-1 vs IVR 400 micrograms.kg-1 or DEC 6 mg.kg-1 alone, for treatment of Wuchereria bancrofti carriers. Of the 57 treated male patients in whom microfilaremia (mf) densities ranged from 22 to 4,709 mf/ml, three groups of 19 were randomly selected, and allocated to one of the three treatments. Twelve months after treatment 37%, 16% and 16% of patients were mf negative in groups DEC, IVR and IVR plus DEC respectively. Mf percent return to pretreatment level was significantly lower in the group IVR + DEC (1.9%) than for DEC 6 (14.7%) or IVR 400 (11.6%). Antigenemia percent return to pretreatment level was lower in the groups IVR + DEC or DEC 6 than for IVR 400. The combination IVR + DEC proved to be the most effective on macrofilariae and microfilariae (antigenemia and mf negative patients). The combination will be a very powerful tool for control of lymphatic filariasis. An annual filariasis day could be the most cost-effective strategy for administration of the drugs.

To demonstrate the liver profile abnormalities in jaundiced falciparum malaria patients and to determine whether jaundice was associated with other complications in falciparum malaria, 390 patients with acute falciparum malaria were studied. 124 patients were jaundiced and the others were non-jaundiced. Hyperbilirubinemia (total serum bilirubin 3 to 64 mg/dl) was found in jaundiced patients predominantly as unconjugated bilirubin. Asparatate amino-transferase and alanine minotransferase were significantly higher in jaundiced patients (p < 0.01). There was a slight decrease of serum albumin in jaundiced malaria. The complications in jaundiced patients included cerebral malaria (n = 10), acute renal failure (n = 12), pulmonary edema (n = 3), shock (n = 3), and other severe malarial complications (n = 43). Jaundice was associated with cerebral malaria (p < 0.05), acute renal failure (p < 0.01), and hyperparasitemia (p < 0.01). After successful treatment, liver profile returned to normal within a few weeks. We found that jaundiced malaria patients had transient liver profile impairment which indicated predominantly hemolysis rather than liver damage; complications were more frequent in jaundiced patients.

Subtherapeutic doses of chloroquine (CQ) are considered to promote development of Plasmodium falciparum resistance but little is actually known about the drug levels in the population in endemic areas. We have therefore measured blood concentrations of CQ in Tanzanian schoolchildren and related these to parasite microscopy. A total of 163 children (median age 11 years) in a suburb outside Dar es Salaam were followed during four weeks. Thick and thin blood films were obtained once weekly. Parasites were counted in 200 visual fields. CQ and desethyl-chloroquine (DECQ) were determined with HPLC in 100 microliters of capillary blood. During the study P. falciparum trophozoites were detected in a mean of 78% of the children, P. falciparum gametocytes in 7.7% and P. malariae parasites in a mean of 13%. The cumulative prevalence of P. falciparum trophozoites and P. malariae parasites was 96% and 28% respectively. On day 0 and day 28, CQ was found in 78% and 80% of the children and DECQ in 21% and 31% of them. A total of 19% of all children had a verified CQ intake during the study and 35% had probably taken CQ. With a few exceptions (9% had CQ concentrations > 100 nmol/l) drug levels were not sufficient to affect parasites with a reduced CQ susceptibility but could possibly promote development of resistance by eradicating the most susceptibility part of the parasite population.

Ivermectin affected the motility of Litomosoides carinii microfilariae in vitro in a dose dependent manner but did not completely immobilize the larvae and had no lethal effects when tested up to a concentration of 1000 ng/ml. However, killing of microfilariae was induced by ivermectin in vitro in the presence of spleen cells of Mastomys coucha or rats within 14 h. Optimum effects occurred at drug levels of 10-100 ng ivermectin/ml. Addition of infection serum led to increased cytotoxicity when compared with normal serum. Pretreatment in vitro of L. carinii microfilariae with ivermectin in cell-free medium and subsequent exposure to spleen cells caused also cytotoxic effects which appeared to be accelerated in comparison with simultaneous exposure of microfilariae to ivermectin and cells. Pretreated microfilariae, injected intravenously into naive M. coucha were rapidly eliminated from the blood of the recipients. These results suggest that the microfilariae become altered by the drug and thus susceptible to cell-mediated cytotoxic effects. Cytotoxicity did not depend on the attachment of cells to L. carinii microfilariae and was also induced when targets and effector cells were separated by membranes impermeable for cells. Thus ivermectin-induced cellular cytotoxicity to L. carinii microfilariae is at least partly mediated by soluble factors released by effective cells.

A Taenia crassiceps metacestode cDNA expression library in lambda gt 11 was screened with rabbit antisera to metacestodal T. solium and T. saginata crude extract. Primary clones (121) were identified, and after rescreening and lysogenization in Escherichia coli Y 1089, were tested in Western blot for reactivity with the same antisera. In addition, analyses were performed with rabbit antisera directed towards T. crassiceps and Echinococcus granulosus metacestode crude extract, sera from humans with neurocysticercosis (Mexico) and other important helminth diseases, mice and calves with experimental T. crassiceps and T. saginata infections and normal sera. Of those tested, 22 clones expressing beta-galactosidase fusion proteins (approximately 118-132 kDa) were reactive with IgG antibodies of cysticercotic patients and T. crassiceps infected mice. Of these clones, 11 were also sero-positive with calf-IgG antibodies against T. saginata larvae. None of the 22 clones reacted with IgG antibodies due to human cystic and alveolar echinococcosis, intestinal/hepatic or urinary schistosomiasis, African onchocerciasis or with sera from uninfected controls (man, rabbit, calf and mouse). Of these 22 clones, 15 have been subcloned into the plasmid vectors pGEX-2T (modified) and pT7T3 alpha 19. Expressed glutathione-S-transferase fusion proteins were again tested for sensitivity and specificity by Western blot, and concentrated by affinity chromatography. The nucleotide sequence of the cDNA inserts of 9 clones has been determined in pT7T3 alpha 19 and revealed identity in 4 and 5 clones, respectively.