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*

Afferent loop syndrome(ALS) is caused by obstruction of the afferent loop after subtotal gastrectomy with Billroth II gastrojejunostomy. Prompt diagnosis of ALS is important as perforation of the loop occurs. The aim of this study is to ascertain the value of sonography and CT to diagnose ALS. We describe the radiologic findings in ten patients with ALS. The causes of ALS, established at surgery, included cancer recurrence (n=4), internal hernia(n=4), marginal ulcer (n=1), and development of cancer at the anastomosis site(n=1). Abdominal X-ray and sonography were performed in all cases, upper GI series in five cases and computed tomography in two cases. The dilated afferent loop was detected in only two cases out of ten patients in retrospective review of abdominal X-ray. ALS with recurrence of cancer was diagnosed in three cases by upper GI series. Of the cases that had sonography, the afferent loop was seen in the upper abdomen crossing transversely over the midline in all ten patients. The causes of ALS were predicted on the basis of the sonograms in three of the five cancer patients. In two cases of computed tomography, the dilated afferent loop and recurrent cancer at the remnant stomach were seen. Our experience suggests that the diagnosis of afferent loop syndrome can be made on the basis of the typical anatomic location and shape of the dilated bowel loop in both sonography and computed tomography.
Abdomen ; Afferent Loop Syndrome* ; Diagnosis ; Gastrectomy ; Gastric Bypass ; Gastric Stump ; Gastroenterostomy ; Humans ; Peptic Ulcer ; Recurrence ; Retrospective Studies

PURPOSE: Prostatic tumor induced gene-1 (PTI-1) is a mutated human EF-la and putative prostatic carcinoma tumor-inducing oncogene, that is differently expressed in prostatic cancer and benign prostatic hyperplasia. And, it is more sensitive marker than prostate- specific antigen (PSA) for detecting human prostate cancer in the bloodstream. This study invastigated the expression of PTI-1 in paraffin embedded tissue of prostatic carcinoma, prostatic intraepithelial neoplasia, and benign prostatic hyperplasia using in situ PCR. MATERIALS AND METHODS: we evaluated expression of PTI-1 in prostatic carcinoma with prostatic intraepithelial neoplasia (PIN) of 32 cases, benign hyperplasia of 20 cases, high grade transitional cell carcinoma of 10 cases and colon cancer of 10 cases for control group. Also, the immunohistochemical staining for PSA was performed to comparison with clinical value of PSA. RESULTS: The serum level of PSA was closely related to stage and Gleason score (p < 0.05). However, the results of immunohistochemical stains were variable to stage and Gleason score. PTI-1 using in situ PCR expressed in 50% of prostatic carcinoma, 41% of prostatic intraepithelial neoplasia, 10% of benign hyperplasia and colon cancer (p < 0.05). No expression is observed in transitional cell carcinoma. In prostatic carcinoma, PTI-1 expressed in 43.8% (7/16) of stage II, 50.0% (5/10) of stage III, and 66.7% (4/6) of stage IV (p<0.05). In PIN, expression of PTI-1 was similar to prostatic carcinoma (p<0.05). CONCLUSION: PTI-1 represented a relatively sensitive marker for prostatic carcinoma and PIN, indicator of prostatic carcinoma progression.
Carcinoma, Transitional Cell ; Colonic Neoplasms ; Coloring Agents ; Humans ; Hyperplasia ; Neoplasm Grading ; Oncogenes* ; Paraffin ; Polymerase Chain Reaction* ; Prostatic Hyperplasia* ; Prostatic Intraepithelial Neoplasia* ; Prostatic Neoplasms