Genes encoding sequence-specific DNA binding proteins have been isolated by screening cDNA libraries constructed in rgt11 expression vector with recognition site DNAs. We isolated a rgt11 recombinant human cDNA clone, designated to C2, using a DNA probe consisted of heptamer of the perfect palindrome (PP; GGGGATTCCCC) of enhancer A (Enh A) of HLA dass I promoter. Sequencing analysis showed that this clone contained a partial cDNA homologous to NF-kB2. Lysogenic E. coli containing the C2 was generated and crude cell extract was prepared. Immunoblot using anti-B-galactosidase antibody showed that this lysogenic E. coli expressed B-galactosidase fusion protein. Electrophoretic mobility shift assay (EMSA) and DNase I footprinting assay were done using crude cell extract and their patterns were compared with nuclear protein extracted from an EBV transformed B lymphoblastoid cell line (BLCL). EMSA showed that crude cell extract prepared from E. coli lysogen speci5cally bound to the PP of Enh A region of HLA class I gene. DNase I footprinting assay showed that the binding sequence of this recombinant B-galactosidase fusion protein was identical to that of nuclear protein extracted from a BLCL. Our data indicate that a Agt11 recombinant cDNA clone was isolated from a human cDNA library using the PP of Enh A of the HLA class I promoter and this clone encoded a B-galactosidase fusion protein capable of binding to the PP and belongs to a NF-xB subunit.
Cell Line ; Clone Cells ; Deoxyribonuclease I ; DNA ; DNA, Complementary* ; DNA-Binding Proteins ; Electrophoretic Mobility Shift Assay ; Gene Library ; Genes, MHC Class I ; Herpesvirus 4, Human ; Humans* ; Mass Screening ; Nuclear Proteins

Subcutaneous emphysema is a complication of the pneumoperitoneum necessary to perform laparoscopy and will be seen more often as laparoscopic techniques are applied to a growing number of intraabdominal procedures. We report a case of subcutaneous emphysema and hypercarbia without pneumothorax or pneumomediastinum during laparoscopic cholecystectomy, which was treated by multiple puncture with 18G needle on emphysematous site. The suspected cause is inadvertent subcutaneous insufflation of carbon dioxide through the trocar sites by increased intra-abdominal pressure for the establishment of pneumoperitoneum. Immediate recognition, evaluation, and treatment of subcutaneous emphysema is necessary since this can be life-threatening complication.
Carbon Dioxide ; Cholecystectomy, Laparoscopic* ; Insufflation ; Laparoscopy ; Mediastinal Emphysema ; Needles ; Pneumoperitoneum ; Pneumothorax ; Punctures ; Subcutaneous Emphysema* ; Surgical Instruments

Aneuploidy ; Breast Neoplasms ; Carcinoma, Squamous Cell ; Cell Cycle Checkpoints ; Centrosome ; Colorectal Neoplasms ; Humans ; Phosphotransferases ; Protein-Serine-Threonine Kinases ; Uterine Cervical Neoplasms

Dopamine receptors in the CNS and other several tissues were identified by physiological, biochemical and radioligand binding techniques. But previous morphological and biochemical studies have been unable to charaterize or determine the tissue distribution of dopamine receptor subtypes because no selective ligands are available yet. Furthermore, the cellular distribution of the dopamine receptor subtypes in the rat kidney is not demonstrated well. The present study utilizes specific antibodies to characterize the renal distribution of this dopamine receptor subtype using light microscopic immunohistochemistry in the rat kidney. In the rat kidney, D1 receptor protein was localized to proximal tubule, distal tubule, renal vessels, medullary collecting tubule, juxtaglomerular apparatus(JGA) and glomerulus. And D2 receptor protein was localized to distal tubule, Henle's loop, proximal tubule, medullary collecting tubules, juxtaglomerular apparatus(JGA) and renal vasculature. The D1 and D2 receptors, which present in the central nervous system, are now identified in the rat kidney. There are some differences in receptors expressing sites on the previous radioligand binding and pharmacologic studies, but these results suggest that at least some of the renal dopamine DA1 and DA2 receptors correspond structually to the central dopamine D1 and D2 receptors.
Animals ; Antibodies ; Central Nervous System ; Dopamine* ; Immunohistochemistry* ; Kidney* ; Ligands ; Rats* ; Receptors, Dopamine ; Tissue Distribution

To elucidate a possible prognostic factor, we studied 91 cases of breast carcinoma for the expression of n-tn23 protein using an immunohistochemical method, and compared these results with the known prognostic parameters of the breast carcinoma. The mn23 protein was intensely stained in the cytoplasm and/or the nucleus of carcinoma cells in 82 cases(90.1%). There were two patterns of cytoplasmic staining; heterogeneous pattern and homogeneous pattern. Among the positive cases, 43 cases(47.2%) were heterogeneous while 39 cases(42.8%) were homogeneous. Axillary lymph node metastases(p<0.005) was found more frequently in the heterogeneous pattern group(79.0%) than in the homogeneous pattern group(41.0%). There was no significant correlation between nm23 protein expression and other parameters such as patient age, tumor size, estrogen receptor, histopathologic grade, and p53 overexpression. Although axillary lymph node metastasis was correlated with the disease free status(p<0.0005) and patient survival (p<0.05), they showed no correlation with nin23 expression. Multivariate analysis showed that axillary lymph node metastasis was the only prognostic indicator(p<0.05), and the expression of nm23 protein was of borderline significance. The results suggest that the homogeneous and/or granular cytoplasmic expression of mn23 protein plays a role in the suppression of nodal metastasis in breast carcinoma and might contribute in predicting patient survival.
Neoplasm Metastasis

No abstract available.
Hand* ; Humans

Tumor angiogenesis is a process leading to formation of blood vessels within tumors and is crucial for maintaining a supply of oxygen and nutrients to support tumor growth and metastasis. Vascular endothelial growth factor(VEGF) plays a key role in tumor angiogenesis including induction of endothelial cell proliferation, migration, survival and capillary tube formation. VEGF binds to two distinct receptors on endothelial cells. VEGFR-2 is considered to be the dominant signaling receptor for endothelial cell permeability, proliferation, and differentiation. Bevacizumab(Avastin, Genetech, USA) is a monoclonal antibody against vascular endothelial growth factor. It is used in the treatment of cancer, where it inhibits tumor growth by blocking the formation of new blood vessels. The goal of this study is to identify the anti-tumor effect of Bevacizumab(Avastin) for oral squamous cell carcinoma cell lines. Human squamous cell carcinoma cell line(HN4) was used in this study. We examined the sensitivity of HN4 cell line to Bevacizumab(Avastin) by using in vitro proliferation assays. The results were as follows. 1. In the result of MTT assay according to concentration of Bevacizumab(Avastin), antiproliferative effect for oral squamous cell carcinoma cell lines was observed. 2. The growth curve of cell line showed the gradual growth inhibition of oral squamous cell carcinoma cell lines after exposure of Bevacizumab(Avastin). 3. In the apoptotic index, groups inoculated Bevacizumab(Avastin) were higher than control groups. 4. In condition of serum starvation, VEGFR-2 did not show any detectable autophosphorylation, whereas the addition of VEGF activated the receptor. Suppression of phosphorylated VEGFR-2 and phosphorylated MAPK was observed following treatment with Bevacizumab(Avastin) in a dose-dependent manner. 5. In TEM view, dispersed nuclear membrane, scattered many cytoplasmic vacuoles and localized chromosomal margination after Bevacizumab(Avastin) treatment were observed. These findings suggest that Bevacizumab(Avastin) has the potential to inhibit MAPK pathway in proliferation of oral squamous cell carcinoma cell lines via inhibition of VEGF-dependent tumor growth.
Blood Vessels ; Capillaries ; Carcinoma, Squamous Cell ; Cell Line ; Cytoplasm ; Endothelial Cells ; Humans ; Neoplasm Metastasis ; Nuclear Envelope ; Oxygen ; Permeability ; Starvation ; Vacuoles ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2

No abstract available.
Gestational Age* ; Humans ; Infant*

No abstract available.
Carcinoma, Renal Cell* ; Genes, Tumor Suppressor*

BACKGROUND: Afterload as well as myocardial contractil ity is an important factor for the adequacy of circulation after cardiac surgery . To noninvasively assess alterations in afterload, we evaluated the changes in aortic blood velocity waveform. MATERIAL AND METHOD: Ascending aortic blood flow was measured by continuous wave Doppler echocardiography befo re and after afterload manipulation in eight open-chest dogs. Nitroprusside was administered singly and simultaneously with epinephrine in various combinations. Left atrial pressure as an index of preload was maintained by saline administra tion. RESULT: The infusion of nitrop russide produced dose-dependent decreases in blood pressure and index of systemi c vascular resistance(ISVR) (all p<0.05 vs baseline), which was associated with increases in peak velocity(PV), mean acceleration(MA) and minute distance, and w ith a decrease in acceleration time(all p<0.05 vs baseline). ISVR obtained durin g nitroprusside infusion had a better correlation with both PV(r=-0.60, p=0.001) and MA(r=-0.52, p=0.003) than with velocity time integral(VTI) or the Doppler t ime intervals. The combined infusion of nitroprusside and epinephrine, unless IS VR was elevated, produced synergistic effects on PV, MA and VTI, but these Doppl er indexes tended to diminish with an elevation in afterload. CONCLUSION: Doppler measuremen t of PV and MA in the ascending aorta may be used to noninvasively assess change s in afterload.
Acceleration ; Animals ; Aorta ; Atrial Pressure ; Blood Pressure ; Dogs ; Echocardiography, Doppler ; Epinephrine ; Nitroprusside ; Regional Blood Flow ; Thoracic Surgery