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.
Agrobacterium tumefaciens* ; Agrobacterium* ; Endocytosis ; Genome, Plant ; In Vitro Techniques ; Plant Cells ; Plant Tumors ; Plants ; Plasmids ; Polyethylene Glycols ; Protoplasts* ; Spheroplasts* ; Tobacco* ; Virulence ; Wounds and Injuries

No abstract available.
Hepatitis B Vaccines* ; Hepatitis B* ; Hepatitis* ; Humans ; Infant*

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)
Animals ; Ascitic Fluid ; Autoradiography ; Chromatography, Ion Exchange ; Cytochrome P-450 Enzyme System* ; Cytochromes* ; DEAE-Cellulose ; Electrophoresis ; Enzyme-Linked Immunosorbent Assay ; Fatty Acids ; Female ; Humans ; Hybrid Cells ; Hydrocarbons ; Liver* ; Mice ; Polyethylene Glycols ; Rats* ; Spleen

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.
Actins ; Ankyrins ; Chloroform ; Cholesterol ; Cytoskeleton ; Electrophoresis ; Erythrocyte Membrane* ; Erythrocytes* ; Glycoproteins ; Hemolysis ; Humans* ; Liposomes* ; Membranes ; Molar ; Nucleic Acids ; Octoxynol ; Osmolar Concentration ; Phospholipids ; Serine ; Sodium ; Spectrin ; Sucrose ; Ultracentrifugation

PURPOSE: To evaluate differential points of patterns of clustered microcalcification between malignant(n=17) and benign(n=46) lesions on mammogram MATERIALS AND METHODS: We retrospectively and prospectively evaluated mammograms of surgically confirmed 63 patients showing clustered microcalcifications. Area, density, number, size, shape of calcification were evaluated along with associated mass and parenchymal distortion. RESULTS: Malignant calcifications were more variable in size(14/17, 77% vs 25/46, 53%) and shape(l 1/17, 64. 8% vs 13/46, 28.2%) than benign counterparts. Pepper, fine granular, branching, comma, tadpole and wormiform calcification were observed in malignant lesion with statistical significance. The malignant calcifications showed more faint(12/17, 70.5% vs 23/46, 50%), irregular margin(17/17, 100% vs 19/46, 42%) and they were usually associated with parenchymal distortion(16/17, 94% vs 9/46, 20%) and ill-defined masses(10/17, 58.9% vs 12/46, 26.1%). CONCLUSION: Clustered microcalcifications with variable size and shape, faint or irregular margin, parenchymal distortion, ill-defined masses seen on mammography, suggest malignancy.
Humans ; Larva ; Mammography ; Prospective Studies ; Retrospective Studies

No abstract available.
Nipples* ; Poroma*

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.
Antibodies, Monoclonal ; Brain* ; Cell Membrane ; Chromatography, High Pressure Liquid ; Isoenzymes* ; Phospholipases* ; Second Messenger Systems ; Seminal Vesicle Secretory Proteins ; Seminal Vesicles* ; Transducers ; Type C Phospholipases* ; Uterus*

Organophosphate is known to damage both the peripheral and central nervous system. We report a case of organophosphate-induced peripheral polyneuropathy with myelopathy. A 46 years old woman who had ingested a large amount of insecticide (organophosphate) was transported to our hospital. Following medical treatment, she was transferred to the Department of Rehabilitation Medicine 1 month later. Upon admission to rehabilitation medicine, the patient was quadriplegic with markedly decreased muscle tone and strength. Electrodiagnostic examination revealed low amplitude of sensory nerve action potential (SNAP), unevokable compound muscle action potential in distal muscles and abnormal spontaneous activities with needle electromyography, which were compatible with peripheral polyneuropathy. Three months later, motor and sensory function of upper extremities were normalized. The muscle tone of lower extremity increased to Ashworth grade II. Follow-up electrodiagnostic examination revealed normalization of SNAP and disappearance of spontaneous activities, but somatosensory evoked potential which were initially normal, revealed prolonged P40 latencies in the lower extremities. These electrophysiological findings were thought to result from the spinal cord lesion and correlated with clinical findings. We diagnosed the patient as peripheral polyneuropathy with delayed myelopathy induced by organophosphate.
Action Potentials ; Central Nervous System ; Electromyography ; Evoked Potentials, Somatosensory ; Female ; Follow-Up Studies ; Humans ; Lower Extremity ; Middle Aged ; Muscle Hypotonia ; Muscles ; Needles ; Polyneuropathies* ; Rehabilitation ; Sensation ; Spinal Cord ; Spinal Cord Diseases* ; Upper Extremity

No abstract available.
Kidney Transplantation* ; Pregnancy*

PURPOSE: Nutritional as well as genetic and hormonal factors play an important role in the bone mineralization during childhood and adolescence. There are several physical and metabolic changes in obese children, and these changes may influence on the mineralization of the skeleton. The studies about bone mineralization of obese children are rare and contradictory. This study was performed to evaluate the influence of childhood obesity on bone mineral density(BMD). METHODS: The BMD of 49 obese and 41 non-obese children were measured at lumbar spines(L2-L4) using dual energy X-ray bone absorptiometry. Then, the results were assessed and compared according to the degree of obesity and pubertal sex maturation. RESULTS: There were no significant differences in BMD between obese children and non-obese children(0.87+/-.19 g/cm2 vs 0.81+/-.13 g/cm2). BMD increased according to the Tanner' pubertal staging, and the most marked increment was observed at overt puberty. No sex difference in BMD was seen in both obese and non-obese children. BMD was highly correlated with age, height, weight and body mass index(BMI), but there was no significant correlation between BMD and osteocalcin. CONCLUSION: BMD of obese children was not significantly different from that of non-obese children, and BMD also was not changed according to the degree of obesity. These findings suggest that BMD is not influenced by obesity in children.
Adolescent ; Bone Density* ; Calcification, Physiologic ; Child* ; Humans ; Obesity* ; Osteocalcin ; Pediatric Obesity ; Puberty ; Sex Characteristics ; Sexual Maturation ; Skeleton