Serological studies on toxoplasmosis were conducted with rabbits sera that were immunized with RH strain or infected with Beverley strain of Toxoplasma gondii. Complement fixation tests, agar-gel double diffusion tests and agar-gel immunoelectrophoresis were performed. Toxoplasma crude antigen was prepared from the organisms in mice peritoneal fluids, which were infected with RH strain of Toxoplasma gondii. The organisms were suspended in saline volume originally exudated and counted in hemocytometer for purity of the organisms over 99 per cent. These suspended organisms were prepared by sonication, and the solution was centrifuged for 30 min. at 10,000 rpm in 4C. These supernatant fluids were used as crude antigen. On the other hand, purified antigens were fractionated on Sephadex G-200 gel filtration. A Sephadex G-200 column, 80 by 4 cm, equilibrated with Tris-HCl-(0.1 M)-NaCl (1.0 M) buffer, pH 8.0 was used. The eluate fractions were collected in 3 ml per hour and the absorbance at 280 nm was measured with a Beckman Du-2 spectrophotometer. Each tube is pooled into 6 fractions by protein density graph. For immunization of rabbits, crude antigen of RH strain was emulsified with an equal amount of incomplete Freund's adjuvant and l ml of mixture was injected subcutaneously into them once a week for 5 successive weeks. Antisera were obtained at an interval of a week, beginning the first week after the last immunization, while several rabbits were infected with Beverley strain of Toxoplasma gondii by inoculating about 200 cysts and antisera were obtained from them serially at a week interval. The results were as follows: The sera from the rabbits immunized with the RH strain or infected with Beverley strain of Toxoplasma gondii againist the crude antigen showed the first positive reactions in 2 or 3 weeks after the administration or immunization in complement fixation tests. Maximum titers appeared in 4 or 5 weeks after immunization with RH strain and in 7 or 9 weeks after infection with Beverley strain respectively. Complement fixation tests showed the positive reactions in the rabbits sera immunized with RH strain against the purified antigens II, III, IV, V and VI: moreover, antigen IV fraction showed the highest titer. On the other hand in the rabbits sera infected with Beverley strain against the purifed antigens II, III and IV fractions showed the positive reaction; especially, antigen fraction IV showed the highest titer. In immuno-diffusion tests, the sera from the rabbits immunized with RH strain and infected with Beverley strain, against the crude antigen appeared the precipitin bands 2 weeks after the immunization or infection. And the former showed the 2 or 5 precipitin bands after 5-8 weeks and the latter showed the l or 2 precipitin bands after 6 weeks. The sera from the rabbits immunized with RH strain against the purified antigens II, III, IV,V and VI showed the precipitin bands, and the sera from the rabbits infected with Beverley strain against the purified antigens II, III and IV showed the precipitin bands in the immuno-diffusion tests. Especially antigen IV was the strongest reaction against the sera from RH strain and Beverley strain. In agar-gel immunoelectrophoresis, the immunized sera against the crude antigen showed 8 arcs. But the infected sera against the crude antigen showed 4 or 5 arcs. The immunized sera against the fractionated antigens II, III, IV, V, VI showed arcs, but against the fractionated antigen IV showed 6 arcs and in the antigens II, III, V, VI showed l or 2 arcs only. On the other hand, the infected sera against the fractionated antigens IV showed 4 arcs, II and III showed the l arcs, which was the most weak of all.
parasitology-protozoa-Toxoplasma gondii ; toxoplasmosis ; rabbit ; immunology ; electrophoresis

In an attempt to investigate the sensitivity of immunodiagnosis in cats experimentally infected with Paragonimus westermani, agar-gel precipitin reaction were studied. Metacercariae of P. westermani were administered to cats in various doses(2-100 metacercariae per cat) and antisera were obtained at an interval of a week. Precipitin bands appeared in homologous antigen-antibody in experimental paragonimiasis between 3 and 5 weeks after infection in all the cats. Almost all the cases in which a large number of worms were detected, showed strong reactions as revealed by deeply stained bands. Precipitin reactions did not necessarily parallel with the number of worms detected. This may be attributable to the individual difference of a cat's conditions. Very weak precipitin reactions were noticed between Clonorchis antigen and Paragonimus antisera of cats, but no reactions were noticed between Paragonimus antigen and Clonorchis antisera of cats or rabbits.
parasitology-helminth-trematoda ; paragonimiasis ; Paragonimus westermani ; immunology ; Clonorchis sinensis ; cat ; rabbit

No Abstract Available.
Amblyopia*

In an attempt to investigate the antigen-antibody relations and the value of immunodiagnosis for several helminths, Ouchterlony tests and immunoelectrophoreses were carried out. Taenia saginata, Cysticercus sp. of cestodes, Clonorchis sinensis, Fasciola hepatica and Paramphistomum sp. of trematodes,and Ascaris suum of nematodes were used as antigens. On the other hand, antisera were obtained by injecting 0.5 ml each of the above antigens and the same amount of complete Freund's adjuvant into rabbits ten times at an interval of one week. The result obtained in this study are as follows: A larger number of precipitin arcs were demonstrated in homologous antigen-antibody reactions than in heterologous antigen-antibody reactions both in Ouchterlony tests and immunoelectrophoreses. Gross reactions were observed between the different species of the same class, but no cross reactions were noticed when the classes were different with one or two exceptions, such as between T. saginata, F. hepatica and A. suum. In A. suum, the difference between the male and female was more distinct in Ouchterlony test and immunoelectrophoresis than in the examination of organs such as genital organ and coeliac fluid. Immunoelectrophoresis revealed specific arcs and higher sensitive reaction than Ouchterlony test, and was considered to be a more valuable method for identifing species and immunological diagnosis.
parasitology-helminth-trematoda ; cestoda ; nematoda ; immunoelectrophoresis ; Taenia saginata ; Cysticercus ; Clonorchis sinensis ; Fasciola hepatica ; Paramphistomum sp. ; Ascaris suum ; antigen ; immunology

In an attempt to investigate the specific antigenic substance of Fasciola hepatica, Ouchterlony tests and immunoelectrophoretic analyses were carried out. Crude Fasciola antigen was prepared and fractionated by Sephadex G-200 column to Antigen I, II and III according to protein content. Crude antigens of Paragonimus westermani, Clonorchis sinensis and Paramphistomum sp. were also prepared for control and absorption study. Antiserum was prepared by injecting 0.5 ml of crude Fasciola antigen with same amount of complete Freund's adjuvant in rabbits, 10 times at an interval of l week. The results obtained in this study were as follows: Crude Fasciola antigen reacted with antiserum with 9 precipitin bands by Ouchterlony test and with 11 bands by immunoelectrophoresis. Cross reaction was observed between Paragonimus, Clonorchis and Paramphistomum antigens and anti-Fasciola rabbit serum respectively. By Ouchterlony test, 3-4 cross reacting bands were found. Anti-Fasciola sera which were absorbed with respective Paragonimus, Clonorchis and Paramphistomum antigens, reacted with Fasciola crude antigen. Ouchterlony test gave 5-6 precipitin bands. Further reaction between Fasciola antigen and antiserum absorbed with the above 3 antigens concomitantly gave 5 precipitin bands by Ouchterlony test and 7 bands by immunoelectrophoretic analyses. Fractionated Fasciola antigens (Antigens I, II and III) reacted with anti-Fasciola rabbit serum in immunoelectrophoresis. Antigen I, II and III gave 2, 3 and 5 precipitin bands respectively. Anti-Fasciola rabbit serum which was absorbed with 3 trematodes antigens gave, by immunoelectrophoresis, 1 band with Antigen I, 2 bands with Antigen II and III of Fasciola hepatica. From the above results, it is concluded that Fasciola hepatica possessed the specific antigenic substance not cross-reacted with other trematodes.
parasitology-helminth-trematoda ; Fasciola hepatica ; Paragonimus westermani ; Clonorchis sinensis ; Paramphistomum sp. ; antigen ; immunology ; electrophoresis

Mutations in the p53 gene occur during the development of colorectal carcinomas, and play an important role in the conversion of adenoma into carcinoma. To detect the p53 gene mutation and its pattern of expression in colorectal carcinomas, polymerase chain reaction for exons 5, 6, 7, and 8, recombinant gene cloning, and automated DNA sequencing were performed with 30 fresh colorectal carcinomas. Each tissue was also analyzed by immunohistochemical staining for p53 protein. p53 protein was detected in 25 of 30 (83.3%) colorectal carcinomas by immunohistochemical study. p53 mutation was detected in 4 of 30 (13.3%) colorectal carcinomas. The distribution of these mutations among these exons investigated was as follows: Three mutations in exon 5 (66.7%) and 1 mutation in exon 7 (33.3%). One case with mutation in exon 5 had mutations at three different codons. Mutations in exon 5 were found at codon 153 (GGG to AGG: Gly to Arg), 170 (TGC to GGC: Cys to Gly), 186 (CTA to TTA: silent mutation), 158 (GCG to ACG: Ala to Thr), and 176 (ACG to ATG: Thr to Met). Mutation in exon 7 was found at codon 248 (AGG to AGA: silent mutation). Four of them were missense mutations. Two of 6 mutations were silent mutations. Five transition mutations and 1 transversion mutation were also detected. All cases with mutations by automated DNA sequencing showed positive p53 protein immunohistochemical stainining. In conclusion, p53 gene mutation was detected in 4 of 30 (13.3%) colorectal carcinomas, located in codon 153, 158, 170, 176, and 186 of exon 5 and codon 248 of exon 7. Further studies are needed to evaluate the significance of the codon 153 mutation which was not recognized in other studies on colorectal carcinomas.
Adenoma ; Clone Cells ; Cloning, Organism ; Codon ; Colorectal Neoplasms* ; DNA* ; Exons ; Genes, p53* ; Mutation, Missense ; Polymerase Chain Reaction ; Sequence Analysis, DNA*

In order to find out a valuable control measure for soil-transmitted parasties, the infectivity in mice of Ascaris eggs irradiated with Cobalt(60) were examined. The results were summarised as follows. In vitro, Ascaris eggs irradiated with larger doses of Cobalt(60) developed poorly, and no difference was found between fresh eggs and those cultured for a few days. Ascaris eggs irradiated with doses of 200,000 rad. developed at the rate of 90 percent after 4 weeks, whereas those irradiated with 1,000,000 rad. developed 28 percent. Ascaris eggs irradiated with Cobalt(60) after 2 weeks of culture were poor in development compared with those of 4 week culture. Eggs cultured for 5 weeks showed weaker infectivity in mice than those cultured for 8 weeks. In the control groups, the infectivity in mice of Ascaris eggs was remained the same between 5 and 8 weeks. The minimum dose of Cobalt(60) irradiation effective for preventing infectivity in mice was estimated to be 200,000 rad.
parasiotology ; radiology ; prevention ; Ascaris suum ; nematode ; Cobalt(60) ; irradiation

Indirect fluorescent antibody tests were performed with sera of paragonimiasis patients and skin test positive sera against Paragonimus antigen. Paragonimus antigen was prepared from lyophilized adult worms of P. westermani by defatting with ethyl-ether before extracting with barbital buffered saline. Preparation of Paragonimus antigen for the indirect fluorescent antibody test was based upon Sato's method used for sero-diagnosis of anisakiasis, with Sephadex G-25 instead of Sepharose 4B. The results were as follows: The indirect fluorescent antibody titers of paragonimiasis patient's sera ranged from 1:64 to 1:512, whereas the control sera showed titers of less than 1:16. As controls, Clonorchis patient's sera and parasite-free healthy human sera were used. In indirect fluorescent antibody tests, the skin test positive human sera against Paragonimus antigen showed a positive rate of 41.5 per cent in the case of titers more than 1:40. On the other hand, complement fixation tests on the same sera showed a positive rate of 32.5 per cent in the case of titers more than 1:20.
parasitology-helminth-trematoda ; paragonimiasis ; Paragonimus westermani ; diagnosis ; indirect fluorescent antibody tests ; serum ; ethyl-ether

No abstract available.
Hematuria* ; Humans

No Abstract available.
Infection Control*