Inositol 1,4,5-triphosphate(InsP,) is a second messenger for obilizing intracellular Cal'. It can be dephosphorylated by soluble and particulate forms on InsP, 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate(InsP,) by InsP, 3-kinase. These enzymes represent possible targets for the regulation of the InsP,AnsP. signal. InsP, 3-kinase which catalyses th ATP-dependent phosphorylation of InsP, was purified from bovine brain tissue. All operation were carried out at 41C. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of InsP, 3-kinase, I and U were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were 1, 0.6 and 11, 4.8 nM/min/mg, and folds
Brain* ; Catalysis ; Chromatography ; Chromatography, DEAE-Cellulose ; Chromatography, High Pressure Liquid ; DEAE-Cellulose ; Inositol* ; Isoenzymes ; Phosphorylation ; Phosphotransferases* ; Polyethylene Glycols ; Second Messenger Systems

No abstract available.
Menisci, Tibial*

No abstract available.

PURPOSE: Na+-K+ ATPase Activity has beem estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characterist-Isc of T1-201 is known to be simiIar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive Na+-K+ ATPase activity using T1-201 and compare with that using Rb-86. MATERIALS AND METHODS: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular Na+-K+ ATPase activity. Na+-K+ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. RESULTS: Na+-K+ ATPase ase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 50-70% or 28% increases from baseline activity, respectively. CONCLUSION: These results suggests that T1-201 could replace Rb-86 in measurement of ouabain sensititive Na+-K+ ATPase activity in vitro. High level of glucose concentration decreased cellular Na+-K+ ATPase activity, but insulin or PMA increased it.
Adenosine Triphosphatases* ; Animals ; Aorta ; Cardiac Glycosides ; Glucose ; Humans ; Insulin ; Membranes* ; Myocytes, Smooth Muscle ; Myristic Acid ; Ouabain ; Potassium ; Rats ; Umbilical Arteries

PURPOSE: Direct injection of doxorubicin into the eyelids results in permanent loss of muscle fiber and it is considered an attractive nonsurgical method in essential blepharospasm therapy. However, necrosis of skin overlying orbicularis oculi muscle is the most serious side effect of this therapy. The purpose of this study is to determine the effect of doxorubicin delivered by osmotic pump which release doxorubicin slowly, and to evaluate the degree of overlying skin injury following chemical myectomy. METHODS: Thirty three rabbits were assigned to three groups according to the doxorubicin concentration. The first group received direct injections of 0.5 mg doxorubicin diluted in 0.1 ml of saline in the right lower eyelid. and osmotic pump was inserted into the left lower eyelid which contained 0.5 mg doxorubicin in 0.1 ml of saline. The second group received 1 mg doxorubicin and the third group received 2 mg doxorubicin. Eight weeks after injection, the eyelids were assessed for the degree of muscle fiber loss microscopically. For the evaluation of functional change of muscle, an EMG study was carried out. RESULTS: Skin necrosis developed in all rabbits except for one which received injection of 0.5 mg doxorubicin. Skin necrosis appeared earlier in the direct injection group. The duration of skin necrosis was shortened at lower concentrations(0.5 mg, 1.0 mg) with a pump delivery(P<0.05). But there was no statistical differences in the 2.0 mg concentration. The size of necrosis was much smaller in rabbits using pump delivery than those of direct injection group in high doxorubicn concentrations(1.0 mg, 2.0 mg)(P<0.05). The total size of muscle fiber was decreased after a doxorubicin injection. There was no statistical difference between the direct injection group and the pump group(P<0.05). The similar effect on the muscle was noted regardless of the slow release of the doxorubicin into the muscle. Light microscopic study demonstrated destructive change of muscle and it was replaced by connective tissues. Electron microscopic study showed destruction of micro-architecture of muscle fibers. Functionally, in EMG study, there was no motor activity in the injection area. But some motor unit potentials appeared in the periphery of skin necrosis site. CONCLUSIONS: These results suggest that the osmotic pump may be used as an effective adjuvant in preventing skin necrosis in blepharospasm treatment.
Blepharospasm ; Connective Tissue ; Doxorubicin* ; Eyelids* ; Motor Activity ; Necrosis* ; Rabbits ; Skin*

The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.
Base Sequence* ; DNA, Ribosomal* ; Nucleotides ; Phylogeny ; RNA, Ribosomal, 5.8S ; Trichoderma*

No abstract available.
Collagen* ; Heparin* ; Mesangial Cells*

No abstract available.
Collagen* ; Glucose* ; Insulin* ; Mesangial Cells* ; Somatostatin*

Cervical intraepithelial neoplasia in human consist of dysplasia of various developmental stages and squamous cell carcinoma in situ of various types. These lesions can be diagnosed cytologically on cervico-vaginal smears, although the diagnostic reproducibility is limited. To obtain the objects morphologic distinction between normal squamous epithelial cell in different maturation, different stages of dysplastic cells and varieties of in situ carcinoma cells, Kontron IBAS-1 imaging analyzer was applied for the measurement of nuclear and cytoplasmic areas of each categorised cells. The followings are results obtained: 1) Nuclear and cytoplasmic areas of superfical (36.9 micrometer2, 2319.9 micrometer2) intermedicate (45.7 micrometer2, 2989.7 micrometer2) and parabasal cells (50.8 micrometer2, 432.7 micrometer2) of normal squamous epithelium origin are mostly distinctive between cell types. However, cytoplasmic areas of both superficial and intermediate cells and nuclear areas of both intermediate and parabasal cells are not significantly different. 2) Normal squamous cells and various dysplastic cells show obvious difference in their nuclear and cytoplasmic areas, while difference between cytoplasmic areas of both parabasal (432.7 micrometer2) and severe dysplastic cells (409.7 micrometer2) are not statistically significant. 3) No statistical difference is observed in between nuclear areas of both moderate dysplastic (112.3 micrometer2) and severe dysplastic cell (117.6 micrometer2). 4) Varieties of carcinoma in situ cells and severe dysplastic cells are in difference in their nuclear and cytoplastic areas, whereas nuclear areas from both in situ carcinoma cells (95.3 micrometer2) of large cell type and severe dysplasia (117.6 micrometer2) are not distinctive. The results lead the author to consider that the morphometric analysis for various parameters of cell constituents are of value in making objective distinction between cells from cervical intraepithelial neoplasia in human.
Humans

The frozen section technique is a means of intraoperative pathological diagnosis, and a procedure of great value to the surgeon. This method should be accurate, rapid and reliable. This method serves useful purposes, such as determining the presence of tumor, its type (especially whether it is benign or malignant), the adequacy of a biopsy of a suspected lesion, and the conditions of the surgical margins. But, it bears many disadvantages, the most of which is the danger of incorrect diagnosis. We studied the indications, the limitations, and the accuracy of the frozen section method and the materials studies was total of frozen section diagnosis of 809 cases was 98.1% with 0.5% of false negative, 0% of false positive 0.5% of incorrect histological diagnosis or grading errors, and 0.9% of deferred cases. The tissues submitted were lymph node, gastrointestinal tract, skin subcutaneous tissues in decreasing oder of frequency. The false positive case is not present, while the false negative cases were 4.
Biopsy ; Diagnosis ; Frozen Sections* ; Gastrointestinal Tract ; Lymph Nodes ; Methods ; Pathology, Surgical* ; Skin ; Subcutaneous Tissue