Isolation of cytotoxicity-potentiating substances from red ginseng / 대한암학회지

No abstract available.

The wormicidal substance of fresh water fishes on Clonorchis sinensis V. Purification and chemical characterization of clonorchicidal substance from epidermal mucus of Cyprinus carpio

As a series of studies to clarify clonorchicidal substances in body surface mucus of some fresh-water fishes, the substance in the epidermal mucus of Cyprinus carpio was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature. Wormicidal trial was done in vitro, and the results obtained are summarized as follows: The mucus was extracted by ethyl ether and separated into 4 fractions by column chromatography using benzene as solvent. The second fraction with yellowish red colour among them showed the strongest clonorchicidal effect. The yellowish red fraction obtained by column chromatography was then fractionated into 6 spots by thin layer chromatography with petrol. ether/chloroform(30/70, v/v), and the Rf. 0.714 spot among the 6 spots showed the strongest effect. The Rf. 0.714 spot was further fractionated into 6 spots by thin layer chromatography with benzene/acetone (90/10, v/v), and the Rf. 0.800 spot among the later 6 spots revealed the strongest effect. The Rf. 0.800 spot was chromatographed on column with benzene and 2 fractions were obtained. The second fraction of light brown colour represented the final purified fraction. By these purification procedures, clonorchicidal substance was purified 15-fold with 0.03 percent yield from the mucus of C. carpio, and 10mg of the final fraction killed the cercaria in 26 min, the metacercaria in 115 min, and the adult in 443 min. Infra red and nuclear magnetic resonance spectrometric analysis of the purified substance revealed that the substance belongs to an ethyl ester of unsaturated fatty acid with 2 double bonds, 15 methylene groups and l methyl group.

The wormicidal substance of fresh water fishes on Clonorchis sinensis IV. Preliminary research on the wormicidal substance from mucus of Cyprinus carpio nudus

As a series of studies on the suitability as a second intermediate host of Clonorchis sinensis, the wormicidal effects of mucus from epidermis of Cyprinus carpio nudus were observed. The results obtained were summarized as follows: On the silica gel thin layer chromatography with the greenish yellow supernatant in acetone:benzene/10:90 as carrier, seven spots were observed in iodine chamber. The spot of Rf. 0.225 value among them disclosed the strongest wormicidal effect on the cercaria, the excysted metacercaria, and the adult of C. sinensis. Meanwhile, no seasonal variations were found in Rf. values and effect of the wormicidal fractions. The wormicidal materials in epidermal mucus of C. carpio nudus and Carassius carassius were compared in their effect, and the former was more effective killing substance than the latter. It seemed that C. carpio nudus could not be proper intermediate host of C. sinensis according to results of the present study, in addition to our previous observation results.

Experimental Infection of Clonorchis sinensis to Cyprinus carpio nudus

As a part of observation on the suitability as a second intermediate host of Clonorchis sinensis, various infection experiments were tried to Cyprinus carpio nudus. The results obtained were summarized as follows: The cercariae which attempted to contact with Cyprinus carpio nudus in the water were observed under the stereomicroscope. After contact, cercariae began to separate its tails from the bodies at 4 minutes increasingly, and then the number ranged to 80 percent at 13 minutes after the encounter. But very few cercariae could actually invade into the epidermis of the fish. When Cyprinus carpio nudus were exposed to a number of cercariae in the beaker, only a few cercariae could invade through the epidermis. Most of the invaded cercariae were killed before forming the cyst. Also rare encysted cercariae were found to be dead within 48 hours. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in the indoor aquarium. When observed after 12 days, to determine the susceptibility, no metacercaria was found infected. Cyprinus carpio nudus were reared with Parafossarulus manchouricus shedding numerous cercariae in Kimhae lake for 2 months in summer of 1982, in order to check the natural infection. No metacercaria of Clonorchis sinensis was found from the fish. In the cases of control fish, Pseudorasbora parva, numerous cercariae were inspected on the course of invasion through epidermis of the fish under the stereomicroscope. And many metacercariae were also found from the fish whenever they were kept in beaker, indoor aquarium, or the lake for natural infection. The results suggested that Cyprinus carpio nudus might not be proper intermediate host of Clonorchis sinensis.

Localization and isolation of anthelmintic activity on Clonorchis sinensis from stem cortex of Machilis thunbergii

The present work was to localize and isolate the anthelmintic substance from stem cortex of Machilis thunbergii which grows in all parts of the Korea peninsula. The dried M. thunbergii bark was extracted with a solvent mixture (Methanol: Ethyl acetate/1:1) by a Soxhlet extractor at 70C for 24 hours, and then the extract was separated into solid matter (negative wormicidal) and benzene extract (positive wormicidal) by benzene solvent. The anthelmintic action was observed on the cercaria, the liberated metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with a spot of benzene extract in benzene solvent, nine spots were observed in iodine chamber and the spot of Rf. 0.16 value of them appeared the most strong wormicidal effect on the cercaria and the liberated metacercaria of C. sinensis. Afterward, a 0.3 g colourless solid substance which was equivalent to the Rf. 0.16 value spot was isolated from the benzene extract of 3 kg M. thunbergii stem cortex by means of silica gel column chromatography using benzene carrier, and the adult of C. sinensis was killed within 6 minutes by 0.05 percent solution of the substance.

Clavate Cells Of Epidermis In Cyprinus Carpio Nudus With Reference To Its Defence Activity To Clonorchis Sinensis

No abstract available.

Isolation of anthelmintic substance on Clonorchis sinensis from tuber of Bletilla striata

The present work was done to isolate the anthelmintic substance from the tuber of Bletilla striata which grows Gangweon-do of the Korea peninsula. The dried B. striata tuber was extracted with ethyl ether by a Soxhlet extractor at 40C for 24 hours, and then the extract was separated into insoluble material (negative wormicidal) and soluble material (positive wormicidal) by petroleum ether solvent. The wormicidal substance was separated in the silica gel column chromatography from the petroleum ether extract, using benzene (90 percent) and acetone (10 percent) as a solvent. The anthelmintic activity was observed in vitro on the cercaria, the excysted metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with the wormicidal substance in benzene:acetone/90:10, five spots were observed in iodine chamber and the spot of Rf. 0.18 value of them appeared the most strong wormicidal effect on the cercaria of C. sinensis. Afterward, a substance which was equivalent to the Rf. 0.18 value spot was isolated from the wormicidal substance by means of silica gel column chromatography using 1 part of ethyl acetate and 6 parts of chloroform carrier, and the excysted matacercaria and the adult of C. sinensis were killed within 14 minutes and 128 minutes in the substance.

The wormicidal substances of fresh water fishes on Clonorchis sinensis II. Preliminary research on the wormicidal substances from mucous substances of various fresh water fishes

The present work which was investigated in July 1979, was to observe the wormicidal effects of the external mucous substances of 9 species of fresh water fishes (Cyprinus carpio, Parasilurus asotus, Anguilla japonica, Ophicephalus argus, Carassius carassius; golden crusian carp, Misgurnus anguillicaudatus, Zacco platypus, Pseudorasbora parva and Carassius carassius; crusian carp) on cercaria, liberated metacercaria and adult of Clonorchis sinensis. The mucous substances extracted by ether from the above 9 species of fishes were separated into many spots on the silica gel thin layer chromatography, and divided into many fractions in the silica gel column chromatography which used petroleum ether (30 percent) and chloroform (70 percent) as a solvent. On the silica gel thin layer chromatography of ether extracts from 9 species of fresh water fishes, each of the Rf. values which had wormicidal effect on the cercaria of C. sinensis were different from others; Rf. value of C. carassius (crusian carp) was 0.937, 0.709 in O. argus, 0.612 in A. japonica, 0.576 in P. asotus, 0.451 in C. carpio, 0.701 in Z. platypus, 0.385 in C. carassius(golden crusian carp) and 0.15 in P. parva. Time for wormicide was different from each other, too. It took 14 min. in a case of C. carassius(crusian carp), 25 min. in Z. platypus, 26 min. in C. carassius(golden crusian carp), 28 min. in C. carpio, 30 min. in P. asotus, 35 min. in O. argus, 40 min. P. parva and 180 min. in A. japonica. But any of the spots of M. anguillicaudatus did not show wormicidal effect on the cercaria within 10 hours. In the silica gel column chromatography of ether extracts from 9 species of fresh water fishes, the fractions which had wormicidal effect on the metacercaria of C. sienesis were different from each other; in a case of C. carassius(crusian carp), the first fraction only had wormicidal effect, the 2nd in P. asotus, the 3rd in O. argus, the 4th in C. carassius(golden crusian carp), the 2nd in C. carpio and the 4th in P. parva. Time for wormicide was 40 min. in C. carassius(crusian carp), 52 min. in P. asotus, 74 min. in C. carassius(golden crusian carp), 92 min. in O. argus, 95 min. in C. carpio and 140 min. in P. parva, but any of the fractions of A. japonicas, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 10 hours. The fractions which had wormicidal effect on the metacercaria of C. sinensis had also the same effect on the adult, but it took longer time to kill them. It took 3 hours in a case of C. carassius(crusian carp), 4 hrs. in C. carassius(golden crusian carp), 5 hrs. in P. asotus, 6 hrs. and 6 min. in O. argus, 6 hrs. and 10 min. in C. carpio and half and 6 hrs. in P. parva. But any of the fractions of A. japonica, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 24 hrs.

The wormicidal substances of fresh water fishes on Clonorchis sinensis III. Seasonal variation on the wormicidal substances of mucous substances of Carassius carassius

The epidermal mucous substances of C. carassius which were extracted by ether were separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether (30 percent) and chloroform(70 percent) as a solvent. The wormicidal effects of the spots and the fractions on the cercaria, the liberating metacercaria and the adult of C. sinensis, were observed by month giving attention to ambient water temperature. Among the four spots of the thin 1ayer chromatography of the ether extracts from C. carassius, the uppermost first spots (Rf. 0.924-0965) had a strong wormicidal effect on the cercaria in May, June, July, August and September, and the cercaria which came in contact with the materials was killed in 29-49 min., but in other months (Rf. 0.612-0.752) it was killed only after 50-115 min. In column chromatography of ether extracts from C. carassius, the first fractions which had a wormicidal effect killed the metacercariae within 38-163 min. in May, June, July, August, and September, but it took about 120-458 min. to kill them in other months, meanwhile the wormicidal time of the fractions on the adults was 445-835 min. in May, June, July, August and September, but was 878-2,130 min. in other months. The Rf. value of the uppermost first spot of the thin layer chromatography of ether extrart from the mucous substance of C. carassius which was reared in the aquarium controlled at 23 C was 0.79, and its first fraction of column chromatography killed the metacercaria and the adult of C. sinensis within 315, and 1,850 min. respectively. As summarized above, the wormicidal materials of the epidermal mucous substances of C. carassius had no relation to ambient water temperature, but the secretion period of this wormicidal materials was the same as the liberating period of the cercariae from Parafossarulus manchouricus.

The wormicidal substances of fresh water fishes on Clonorchis sinensis I Preliminary research on the wormicidal substance from mucous substance of Carassius carassius

The present work was to observe the wormicidal effects of the external mucous substance of Carassius carassius and Pseudorasbora parva on cercaria, metacercaria (liberating) and adult of Clonorchis sinensis. The mucous substance which was extracted by ether was separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether(30 percent) and chloroform(70 percent) as solvent. R(f) 0.952 value of the silica gel thin layer chromatography and the first fraction (reddish yellow) frorn the silica gel column chromatography of the ether extracts from C. carassius had the wormicidal effect on all of the developing stages of C. sinensis. Cercaria was killed within 14 minutes, metacercaria within 27 minutes and adult 2.5 hours by the first fraction. But in the normal saline solution (control) and in the other fraction, cercaria and metacercaria were alive more than 24 hours and adult more than 72 hours. The extracts from the mucous substance of P. parva that is well penetrated by cercaria had no wormicidal effect on them.