Table of Content

30 November 1988, Volume 6 Issue 4

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DIAGNOSTIC PROTEINS OF 31/32 KD IN SCHISTOSOMA JAPONICUM: REACTIVITY WITH PATIENT SERA AND MONOCLONAL ANTIBODIES

1988, 6(4):  245-248. 

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Sera of schistosomiasis japonica patients and of S. japonicum-infected mice were tested in immunoblots for their reactivity with adult schistosome proteins. We demonstra-ted the existence in S. japonicum of proteins sharing the diagnostic antigenicity of S. mansoni proteins, with approximately the same molecular weight of 31/32 KD, their reactivity with monoclonal antibodies and association with the schistosome gut. The proteins are, however, different between the two species at least in respect to one epitope and the relative mobility of one component. We confirmed the specificity of S. japonicum proteins by testing with sera of patients with other parasitic diseases. The data add support to the possible application of 31/32 KD schistosome proteins for serodiagnosis of sehistosomiasis Japonka.

SOME ULTRASTRUCTURAL CHARACTERISTICS OF PLASMODIUM YOELII YOELII IN ERYTHROCYTIC SCHIZOGONY AND GAMETOCYTOSIS

1988, 6(4):  249-252. 

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By pretreating Plosmodium yoelii yoelii-infected erythrocytes with saponin combined with glutaraldehyde in hypotonie buffer solution, the process of schizogony such as the formation of bud, apical complex, and trilaminate pellicle of the merozoites were clearly demonstrated under TEM.An organelle, which consisted of several saccules arranged in a parallel fashion together with a cluster of small vesicles, was seen in mature gametocytes near their nucleus. It was supposed to be the Golgi apparatus in terms of its morphological feature.

ABC-ELISA IN THE DETECTION OF SPECIFIC IgE IN PATIENTS WITH HYDATIDOSIS

1988, 6(4):  253-255. 

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A simple and sensitive immunoenzymatic assay-ABC(Avidin-Biotin-HRP Complex). ELISA for detection of anti-hydatid specific IgE antibodies is presented. Sera from 40 patients with hydatidosis tested by both ABC-ELISA and conventional ELISA showed that the former was more sensitive than tne latter, particularly in cases with IgE in lower levels. The positive percentages were 92.5% and 82.5% in s閞um samples diluted 1:100; 75% and 57.5% in 1:200, respectively. The difference of non-specific reactions in 24 cases with cysticercosis cellulosae and 25 cases with allergy was not significant with the two methods. The average levels of total IgE tested by the ABC-ELISA in patients with hydatidosis, cysticercosis, or allergy and in 25 healthy blood donors were of 5 982.96, 3 449.88, l 334.43 and l 150.54 IU/ml respectively, and the deviation from mean square showed differences of statistical significance.

DIAGNOSIS OF FILARIASIS BY INDIRECT FLUORESCENT ANTIBODY TEST WITH SONICATED MICROFILARIAE MALAYI AS ANTIGEN

1988, 6(4):  256-258. 

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Indirect immunofluorescent antibody test (IFA) using sonicated microfilariae of B. malayi as antigen was carried out to detect filariasis since 1983. The microfilariae were first sonicated into fragments 20-30μm long, and then applied to nohecutane-coated slides. Specific fluorescence was seen at the cut ends of the fragmented microfilariae and along the cuticle. Comparison of results obtained with sera collected by venepuncture and eluates of ear lobe blood on filtet paper showed no statistical difference. Hence, the latter could be used instead of the former.This antigen was tested against 114 plasma samples from both bancroftian and ma-layan microfilaraemia cases and 191 normal controls. At the titre of 1:10, the sensitivity was 86.8% and the specificity, 98.4%.In field investigation, when the same titer was considered as positive, the increase in positive rate among population coincided with the positive rate of microfilaraemia and the variation of positive rates ran parallel with the degree of endemicity. It is concluded that the sonicated microfilarial antigen might be useful for the epidemiological investigation and surveillance of filariasis.

ANALYSIS OF THE COMMON ANTIGEN COMPONENT BETWEEN BRVGIA MALAYI ADULTS AND ONCHOCERCA ARMILLATA ADULTS

1988, 6(4):  259-260. 

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The common antigen component between B. malayi and O. armillata was demonstra-ted by cross immunoelectrophoresis(XIE) with antigens from these 2 species of filariae against the sera from patients infected with B. malayt. When the two antigens were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme linked im-munoblotting (ELIB), the component of approximately 43 KD was identified to be the common antigen component.

THE RECONFIRMATION OF THE EXSISTENCE OF LITHOGLYPHOPSIS MODESTUS (GREDLER, 1886) IN HUNAN PROVINCE, CHINA AND ITS ARTIFICIAL INFECTION WITH SCHISTOSOMA JAPONICUM

1988, 6(4):  261-263. 

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Gredler (1886) nominated a species of fresh water snail collected from Hunan Province, China, as Lithoglyphus modestus. Yen (1939) described four different species of Lithoglyphopsis, which had been collected from China and kept in the Natural Museum of Senkenberg, and one of them, L. modsstus, was dofined as the genotype of Lithoglyphpopsis, No further reports concerning the above-mentioned snail species were available since 1939.This paper reconfirms the existence of Lithoglyphopsis modestus in Hunan Province, as they have been found in Hengshan County of the province by the authors. The mo rphology of L. modestus was described in detail. The parasitologic relationship between L. modestus and Schistosoma japonicum was studied and the result showed that L. modestus can not serve as an intermediate host of S. japonicum in spite of the fact that miracidia did invade into the snail.

DETECTION OF ANTIBODIES AGAINST HUMAN HOOKWORMS IN EXPERIMENTALLY INFECTED MICE AND NATURALLY INFECTED HUMANS

1988, 6(4):  264-267. 

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Specific antibodies in the sera of mice experimentally infected with A. duodenale or N. americanus and of hookworm-infected individuals were detected by ELISA using in-fective larva or adult worm antigens.The antibody in mouse model could be detected as early as 7 days after infection with A. duodenale and remained detectable for at least 90 days, with a peak on the 15-30th day. But from 5-45 days after infection with N. americanus, antibody was dete-ctable at day 5 and only lasted 45 days with a peak on the l0th day. It is suggested that the pattern of antibody response may be useful for early diagnosis of hookworm infection and that it may be closely related to the duration of the larvae staying in the host tissues.Sera from 40 patients showed positive rates of 87.5 and 62.5% respectively with adult and larval antigens of A. duodenale. ELISA might be useful immunodiagnostic method for hookworm infection as well as a sero-epidemiological tool for mass examina-tion of ancylostomiasis.

STUDIES ON SURFACE ELECTMCITY OF ENTAMOEBA HISTOLYTICA

1988, 6(4):  268-271. 

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In this paper, the variation of surface electricity of two Entamoeba histolytica isolar es was investigated using the cell electrophoretic technique. By testing the electrophoretic iondition of amoeba in gradient pH and the effect of some enzymes, we provided infor-nations for the physiology and pathogenicity of this parasite.Cell electrophoresis for the two isolates of E. histolytica (isolate A and isolate B) n pH 2.05-8.00 Mcllvaine buffer revealed that EPM (electrophoretic mobility) was negative under the pH gradients of 3.00-8.00, which meant the amoeba cell surfacecar- ried negative charge in this condition. Surface charge densities of isolates A and B were found to be ( + ) 148.3-(-) 6035.9 esu/cm2 and (+ )222.4-(-) 6624.0 esu/cm2 respectively. In addition, except in pH 2.05, the EPMs in Mcllvaine buffer were different between the two amoeba isolates, with significant statistical difference.The enzyme test demonstrated there was no neuraminic acid on the surface of these two amoeba isolates. The surface negative groups of the protozoa were sensitive to trypsin and deoxyribonuclease I.

THE MOUSE MICRONECLEUS TEST FOR MUTAGENICITY OF PYRONARIDINE, CHLOROQUINE, QUINACRINE, PRIMAQUINE, FURAPYRIMIDONE AND PHENTHIOUREZINE

1988, 6(4):  272-274. 

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Four antimalarials, pyronaridine, chloroquine, quinacrine, primaquine, and two an-tifilarials, furapyrimidone, phenthiourezine (I) were administered to groups of Kunming hybrid male mice at ≥LD50 doses. Giemsa stained bone marrow cell smears of the mice were made 24 h after administration and the micronulceated polychromatic erythrocytes pel 1000 polychromatic erythrocytes (MNPCE % ) were counted. The MNPCE %?of each mouse in the six drug groups was 4‰, within the range of the controls, 1.3±1.6‰ (X ± SD), indicating that the six drugs were negative in micronucleus test, having no Structural chromosome damage or damage to the spindle apparatus.

INHIBITORY EFFECTS OF EXTRACTS OF ASCARIS SUUM ON HUMAN BLOOD COAGULATION

1988, 6(4):  275-277. 

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The effects of various soluble extracts of Ascaris suum on human blood coagulation were studied. The extract of whole worm could prolong recalcification time (RT) and kaolin-activated, partial thromboplastin time (KPTT), but did not alter prothrombin time (PT), indicating that the intrinsic pathway of blood coagulation was inhibited by this extract, but the extrinsic pathway was not affected. Whole worm extract inhibited platelet aggregation induced by ADP, but did not influence the one induced by adrenaline. Neither whole worm extract nor body fluid caused fibrinolysis. In KPTT assays with three dif-ferent extracts, cuticle extract exhibited the strongest anticoagulant activity, while whole worm extract and body fluid much less so. These data suggested that anticoagulants of ascaris mainly exist in the cuticle,

STUDIES ON ADENYLATE CYCLASE IN SCHISTOSOMA JAPONICUM

1988, 6(4):  278-281. 

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The activity of adenylate cyclase in Schistosoma japonicum was assayed by radiometric raethod. The 3H-cAMP formed during enzymatic reaction was separated from substrate 3H-ATP by a short Dowex 1×2 (100-200 mesh) 0.5×3 cm column, then the radioactivity of 3H-cAMP was determined by liquid scintillation measurement.The optimum pH of the enzyme activity in S. japonicum homogenate is 7.5. Both male and female worms possess almost the same activity, which increases with the age between 20 to 48 days. The enzyme activity of 40-day-old adult worm was 80000 ± 3000 cpm/pair worm-minute. The activity of adenylate cyclase on the surface membrane of the male worm was one-fold higher than that of the female.3.3× 10-4M of nithrocyamine, metrifonate and niridazole had no effect on the activity of adenylate cyclase in S. japonicum homogenate. 3.3×10-M of furapromidum showed a slight inhibition, whereas saturated pyquiton inhibited 50% activity of the enzyme on the surface membrane of the worm.

APPLICATION OF DOT-IMMUNOBINDING ASSAY IN IMMUNO-DIAGNOSIS OF FALCIPARUM MALARIA

1988, 6(4):  282-284. 

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In this paper, the results of dot-immunobinding assay (DIA) in tne diagnosis of falciparum malaria were reported. Among 53 sera from confirmed falciparum malaria cases from Hainan, 96,2%(51/53) showed positive reactions, while none of the 42 serum samples from blood donors in Shanghai and Ningxia Hui Autonomous Region where malaria was absent was found positive (Table 1). No cross reaction was observed with sera from cases of schistosomiasis, visceral leishmaniasis, paragonimiasis and fasciolopsiasis buski. The positive rate of DIA is higher than that of conventional ELISA(84.9%) or IFA(83.0%). It is recommended as a serological test of choice for the diagnosis of fal-ciparum malaria.

EFFICACY OF COMBINED USE OF PYRONARINE, SULFADOXINE AND PYRIMETHAMINE IN THE TREATMENT OF FALCIPARUM MALARIA ON HAINAN ISLAND

1988, 6(4):  289-291. 

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40 patients of falciparum malaria were treated by pyronaridine 800mg jointly with sulfadoxine 1000mg and pyrimethamine 50mg in 2days in endemic area of chloroquine-resistant falciparum malaria on Hainan Island. All the patients in tne combined treatment group became a febrile in an average of 35 ± 12 h and free from a sexual forms in 40 ± 14 h. No recruescence was seen in 39 patients followed-up 28days after medication, the cure rate being 100%. Only mild side effects were observed in the combined treatment group. Besides, 9 patients of falciparum malaria were treated with piperaquine phospbate l500mg alone for 3 days. The average fever subsidence time and parasite clearance time of the group was 39±l5h and 61±27h, respectively. One patient had recrudescence of parasitaemia and two patients showed high resistant level (RIII) to this drug within 28days after medication, the cure rate being 67%.

EFFICACY OF AMODIAQUINE, FANSIDAR AND THEIR COMBINATION IN THE TREATMENT OF CHLOROQUINE-RESISTANT FALCIPARUM MALARIA

1988, 6(4):  292-295. 

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During 1985-1986, a comparative double blind randomized trial was carried out in Hainan Island where Plasmodium falciparum is resistant to chloroquine.Fifty cases of falciparum malaria were treated with l800mg amodiaquine (AQ)/3 days, with 65.3% cure rate fonly 50% cure rate in the migrant cases). 30.7 hr and 60.3 hr were needed to clear their fever and asexual parasitaemia, respectively. 34.7% of all cases showed RI and RII.Twenty-one cases were treated with Fansidar(FD) at a single dose of l 500mg. The fever clearance time was 56.1 hr and asexual parasitaemia clearance time, 61.1 hr. Among 21 cases, 19 cases were cured, l case showed RI, l case showed S-RI. Fansidar was not good in fever control, but rather good in radical cure. Forty-nine cases were treated with amodiaquine plus fansidar (AFD) and 50 cases were treated with amodiaquine plus sulfadoxine (AS). Fever was controlled in 25.7 hr. and 25.0 hr., and the asexual blood parasites were eliminated in 52.8 hr. and 57.1 hr. respectively. Cure rate was 100% and 97.9% respectively.After drug administration, the rate of gametocytemia was rather high in the 4 groups, with the highest positive rate of 95.2% in FD group.

A COMPUTER PROGRAMME FOR AUTOMATIC RECOGNITION OF SCHISTOSOME EGGS

1988, 6(4):  296-298. 

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The digital image processing system is mainly composed of the IBM Personal Com-. puter with Plug-compatible, PCVISION Frame Grabber. The accuracy rate in recognising schistosome eggs is 88.2% and 85.4% for two different ways of processing stool, i.g. by nylon tissue concentration and Kato-Katz technique. The false positive rate is 1.8% and 2.2%, respectively. The system can not only process and analyse the morphological chara-cteristics of schistosome eggs in the stool but also print out the report or tables and statistical data of the observation results automatically. It is promising to be one of the possible diagnostic methods for schistosomiasis and deserve further investigation,

TRANSECTIONAL SURVEILLANCE IN WEST MUBEI WITH FILARIASIS BASICALLY ERADICATED

1988, 6(4):  299-300. 

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A transectional Surveillance was carried out in 384 villages of 12 counties in west Hubei where filariasis had been basically endicated 2-3 years ago.Among 155 235 persons surveyed in an area with bancroftian filariasis, 107 micro-filaremic patients were found by blood examination, the average microfilaremia rate (MFR) being 0.07%. However, MFR was as high as 1.31 and 1.91% in 2 villages, being slightly above the parameters used for "basical eradication" of filariasis, the highest microfilarial density was 40 mff/120μl and only l out of 2 785 Culex pipiens fatigans dissected was found harbouring 2nd stage larvae. The authors deemed that in such areas, Surveillance should be strengthened.Concurrently another follow-up observation in areas of malayan filariasis with 64 041 persons from 74 villages was carried out. The results showed that only one patient who had never been examined was found positive by blood examination, the average MFR being 0.0016%, the MFD being 7 mff/120μl. Dissection of 2 706 Anopheles sinensis failed to find larval infection. These results suggest that filariasis transmission in this area has been interrupted, the main task in the later Stage of controlling filariasis is to treat those patients with symptoms and signs.