Homogeneity of Phospholipase C of Bovine Uterus and Seminal Vesicle Compared with Brain Isozymes / 영남의대학술지

Phosphoinositide-specific Phospholipase C (PI-PLC) is a second messenger of signal transducer on cell membrane. In the previous study, PLC of bovine brain has been purified three isozymes. In this paper, uterus and seminal vesicle have been purified. Two peaks of PI-PLC activity were resolved when bovine uterus and seminal vesicle proteins were chromatographed on a DEAE and phenyl TSK 5PW HPLC column. Each two peak was compared with PI-PLC I, II and III from bovine brain and we got the retention time on HPLC. The peak fractions with PLC activity were tested homogeneity with brain PLC monoclonal antibodies (Mab). Mab-labeled affigels were bounded in the range of 73.8%~97.5% with PLC I, II and III. Homogeneity of fractions were revealed that DEAE F-1 and phenyl F-1-I were highest level of PLC III in uterus and seminal vesicle and DEAE F-2 and phenyl F-2-I were mixed PLC I and II.

Change of Pulmonary Function after Exercise Loading Test and Preventive Measures on Exercise-induced Asthma in Chlidren

No abstract available.

Purification of Band 3 from the Human Erythrocyte Membrane and its Incorporation into Liposome / 영남의대학술지

Band 3, the predominant 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneously upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane (ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol (1:1 molar ratio) were dissolved in chloroform and the chloroform was removed by rotator evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Production of Monoclonal Antibody to Polychlorinated Biphenyl Induced Cytochrome P-450 LMII in Rat Liver / 영남의대학술지

Cytochrome P-450 (CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated biphenyl (PCB) – induced CP-450 LMII. The spleen cells derived from immunized mice were fused with SP2 myeloma cells using polyethylene glycol (PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterin-thymidine (HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cells(×107) were inoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascetic fluids. Monoclonal antibody (Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and I¹²⁵ labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW:55,000 and 110,000)

Cytological Study of the Introduction of Agrobacterium tumefaciens Spheroplasts into Nicotiana tabacum Protoplasts / 영남의대학술지

Agrobacterium tumefaciens induces cancerous growths called crown galls at wound sites on dicotyledonous plants. A large plasmid called T1 plasmid is responsible for virulence. Upon tumor induction, part of the plasmid, termed T-DNA, becomes integrated into plant genome and its genetic sequences are expressed. These properties allow T1 plasmids to be used as gene vectors in plants. Several in vitro methods for the transfer of T1 plasmid into plant cell have been developed. One of them is the treatment of bacterial spheroplasts and plant protoplasts mixture with polyethylene glycol that is generally used as fusogen in cell-to-cell fusion. Several workers investigated the interaction of bacterial spheroplasts with plant protoplasts in the presence of polyethylene glycol and suggested that the interaction is not fusion but endocytosis. In this report we observed the interaction of Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts by electron microscope. Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts were prepared and mixed in the presence of polyethylene glycol and high pH-high Ca²⁺ buffer. Then the interaction of the spheroplasts with the protoplasts was examined by transmission electron microscope. After the treatment of polyethylene glycol the spheroplasts adhered to the surface of the protoplasts and then they were engulfed by the protoplasts. After the high pH-high Ca²⁺ buffer treatment the engulfed spheroplasts lost their cell integrity. No fusion process was observed. Thus all these observation suggest that the introduction process of Agrobacterium tumefaciens spheroplasts into Nicotiana tabacum protoplasts with the aid of polyethylene glycol is endocytosis.

Environmental Studies on Paragonimiasis in Korea

Environmental studies regarding Paragonimiasis have been done in Sulak-Myun, Kapyung-Goon (I district) and Yongmoon-Myun, Yangpyung-Goon (II district), both of which are considered "active" endemic areas, and in Byukje-Myun, Koyang-Goon (III district) an "arrested" area. 1. Paragonimiasis is still active in the Uhmos River, Sulak-Myun, Kapyung-Goon, where 49% of crayfish contained Paragonimus metacecaria. Snails which are the first intermediate host of Paragonimus westermani were moderately present. In the Oonke River, Yongmoon-Myun, Yangpyung-Goon, crayfish which are the second intermediate host of Paragonimus westermani, were found in every area of the river, but snails were not found in any area of either branch of the river. They were caught in areas which are located below the junction. In the Daeja river, Bykje-Myun, Koyang-Goon, crayfish were found only in the upper stream of the river and snails were not caught in any area of the river. 2. The results described above are attributed to the fact that snails, which have minimal mobility and vitality, were complete1y obliterated by the long, unprecedented drought in 1965. In contrust, crayfish with great mobility and vitality survived. Thus without the snail, the life cycle of Paragonimus western1ani become incomplete, and the clinical disease disappeared. 3. Rainfall is very important in maintaining water flow in the river, a necessary factor for the life cycle of Paragonimus westermani. Therefore we have charted tile amount of rainfall in the Seoul and Choonchun areas in 1965, compared with the average of the past 30 years, in order to ascertain the degree of drought which influnced the Paragonimus westermani hosts and cohabitants. 4. Geographic conditions which are also very important in maintaining water flow in small rivers was investigated. The conclusion was drawn, that a long river has an advantage over a short during drought; a tortuous river provides a better opportunity for miracidia of Paragonimus westermani eggs to migrate to snails, and for Pargonimus cercaria to invade crayfish than a straight rapidly flowing river. 5. We must conclude that man may ingest Paragonimus metacercaria by drinking water from shallow wells and streams in areas endemic to paragonimiasis, when we note that one crayfish of four caught in shallow wells in Kapyung-Goon contained many Paragonimus metacercaria. 6. Other factors are the increase in traffic due to the expanding population in Korean farm areas, and the increase in the use of farm chemicals, which seep into the rivers and reduce the number of snails, crayfish and crabs available as potential hosts. 7. In recent years, the wild fauna in the mountains, one of the important terminal hosts of Paragonimus westermani, have been markedly reduced in number. This may have contributed to decreasing paragonimiasis as directly infective to man. 8. Health education on paragonimiasis, informing the people of the dangers of eating raw or inadequately cooked crayfish and crabs, or those soaked less than 10 days in soysauce, and drinking the water from shallow wells and streams, is demonstrated to have decreased the incidence of this disease. 9. Because of the preceding facts, paragonimiasis in Kyunggi-Prvince is being decreased.