7-Hydroxystaurosporine (UCN-01) induces apoptosis in human colon carcinoma and leukemia cells independently of p53

7-hydroxystaurosporine (UCN-01) is a more selective protein kinase C inhibitor than staurosporine. UCN-01 exhibits antitumor activity in experimental tumor models and is presently in clinical trials. Our study reveals that human myeloblastic leukemia HL60 and K562 and colon carcinoma HT29 cells undergo internucleosomal DNA fragmentation and morphological changes characteristic of apoptosis after UCN-01 treatment. These three cell lines lack functional p53, and K562 and HT29 cells are usually resistant to apoptosis. DNA fragmentation in HT29 and K562 cells occurred after 1 day of treatment while it took less than 4 h in HL60 cells. Cycloheximide prevented UCN-01-induced DNA fragmentation in HT-29 cells, but not in HL60 and K562 cells, suggesting that macromolecular synthesis is selectively required for apoptotic DNA fragmentation in HT29 cells. UCN-01-induced DNA fragmentation was preceded by activation of cyclin B1/cdc2 kinase. Further studies in HL60 cells showed that UCN-01-induced apoptosis was associated with degradation of CPP32, PARP, and lamin B and that the inhibitor of caspases (ICE/CED-3 cysteine proteases), Z-VAD-FMK, and the serine protease inhibitor, DCI, protected HL60 cells from UCN-01-induced DNA fragmentation. However, only DCI and TPCK, but not Z-VAD-FMK, inhibited DNA fragmentation in the HL60 cell-free system, suggesting that serine protease(s) may play a role in the execution phase of apoptosis in HL60 cells treated with UCN-01. Z-VAD-FMK and DCI also inhibited apoptosis in HT29 cells. These data demonstrate that the protein kinase C inhibitor and antitumor agent, UCN-01 is a potent apoptosis inducer in cell lines that are usually resistant to apoptosis and lack p53 and that caspases and probably serine proteases are activated during UCN-01-induced apoptosis.     

Insights into multistep phosphorelay from the crystal structure of the C-terminal HPt domain of ArcB

The histidine-containing phosphotransfer (HPt) domain is a novel protein module with an active histidine residue that mediates phosphotransfer reactions in the two-component signaling systems. A multistep phosphorelay involving the HPt domain has been suggested for these signaling pathways. The crystal structure of the HPt domain of the anaerobic sensor kinase ArcB has been determined at 2.06 A resolution. The domain consists of six alpha helices containing a four-helix bundle-folding. The pattern of sequence similarity of the HPt domains of ArcB and components in other signaling systems can be interpreted in light of the three-dimensional structure and supports the conclusion that the HPt domains have a common structural motif both in prokaryotes and eukaryotes.     

A growth control approach to Class II, Division 1 cases during puberty involving the simultaneous application of maxillary growth restriction and mandibular forward induction

A simple method that used headgear and a functional appliance simultaneously was used for the correction of Class II, Division 1 cases with severe denture base discrepancy. The treatment restricted the forward growth of the maxilla and advanced the mandible. The functional appliance, referred to as the mandibular growth advancer (MGA), advances the mandible progressively with a splint, with the objective of remodeling the condyle and the glenoid fossa in the temporomandibular joint. Functional adaptation was achieved as the muscles that are attached to the mandible adjusted to new positions. In the two cases that illustrate this method, the ANB angle decreased and the Ar-B distance increased over a short period to four and six times the mean Japanese growth rate, respectively. After the correction of the denture-base discrepancy, a multibracket fixed appliance was used for dental alignment, and good skeletal, occlusal relationships and profiles were obtained. Treatment of severe denture-base discrepancy in this manner may reduce the skeletal abnormality, decrease the number of extraction cases, and shorten the subsequent multibracket treatment time. And it may reduce the iatrogenic side effects caused by prolonged mechanotherapy with a fixed appliance.     

Three-dimensional display of the bronchi using helical CT

The purpose of this study was to evaluate the clinical usefulness of three-dimensional (3D) images of the bronchi obtained using helical CT. Thirteen patients with lung cancer, one with tracheal diverticulum, and one with bronchial amyloidosis were examined. The CT scanner employed was the Toshiba Xforce. The helical CT scan cycle consisted of 20 continuous rotations, each requiring 1.5 sec, for a total scanning time of 30 sec. Scans were obtained using a 5-mm X-ray beam width, a 5-mm/1.5 sec couchtop sliding speed, and a 2-mm reconstruction interval. 3D images were reconstructed using a CEMAX VIPstation. The optimal lower and upper threshold CT values for 3D images of the bronchi were -650 and -100 HU, respectively, and 3D images clearly depicted endobronchial lesions. Cartilage crescents were also demonstrated, but longitudinal and circular mucosal folds could not be visualized. In conclusion, 3D images of the bronchi acquired using helical CT were useful in evaluating endobronchial lesions.     

Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae.

Drug-resistance markers for yeast transformation are useful because they can be applied to strains without auxotrophic mutations. However, they are susceptible to technical difficulties, namely lower transformation efficiency and the appearance of drug-resistant mutants without the marker. To avoid these problems, we have constructed a phosphoglycerate kinase (PGK) promoter-driven YAP1 expression cassette, called PGKp-YAP1. Yeast cells containing PGKp-YAP1 were resistant to cycloheximide, a protein synthesis inhibitor, and also to cerulenin, a fatty acid synthesis inhibitor, but not to other drugs tested. The transformation efficiency of PGKp-YAP1 using cerulenin selection was comparable to that using a URA3 auxotrophic marker when low concentrations of cerulenin were used. Non-transformed drug-resistant colonies did appear on the low-concentration cerulenin plates. However, these non-transformed colonies could easily be identified, based on their cycloheximide sensitivity and/or their resistance to aureobasidin A to which the transformants were sensitive. Therefore, the dual drug resistance of PGKp-YAP1 could be used as an effective selection for PGKp-YAP1 recipient cells. The PGKp-YAP1 marker was used to disrupt the LYS2 gene and to transform an industrial yeast strain, indicating that this marker can be used for efficient and reliable gene manipulations in any Saccharomyces cerevisiae strain.     

Development of a distributed sensing technique using Brillouinscattering

This paper reviews the developments of a distributed strain and temperature sensing technique that uses Brillouin scattering in single-mode optical fibers. This technique is based on strain- and temperature-induced changes in the Brillouin frequency shift. Several approaches for measuring the weak Brillouin line are compared     

A new write/erase method to improve the read disturbcharacteristics based on the decay phenomena of stress leakage currentfor flash memories

This paper describes a new write/erase method for flash memory to improve the read disturb characteristics by means of drastically reducing the stress leakage current in the tunnel oxide. This new write/erase operation method is based on the newly discovered three decay characteristics of the stress leakage current. The features of the proposed write/erase method are as follows: 1) the polarity of the additional pulse after applying write/erase pulse is the same as that of the control gate voltage in the read operation; 2) the voltage of the additional pulse is higher than that of a control gate in a read operation, and lower than that of a control gate in a write operation; and 3) an additional pulse is applied to the control gate just after a completion of the write/erase operation. With the proposed write/erase method, the degradation of the read disturb life time after 10⁶ write/erase cycles can be drastically reduced by 50% in comparison with the conventional bipolarity write/erase method used for NAND type flash memory. Furthermore, the degradation can he drastically reduced by 90% in comparison with the conventional unipolarity write/erase method fur NOR-, AND-, and DINOR-type flash memory. This proposed write/erase operation method has superior potential for applications to 256 Mb flash memories and beyond     

A novel therapeutic approach for rectal varices: a case report of rectal varices treated with double balloon-occluded embolotherapy

We present a patient with continuous melena, diagnosed as rectal varices bleeding. She had a history of esophageal varices, which was treated by endoscopic ligation therapy. Eight years after the treatment of esophageal varices, the continuous melena began. Colonoscopic examination showed that the melena was caused by rectal varices, which were so severe that they could not be treated by either endoscopic sclerotherapy or surgical devascularization. Taking into considering the overall risk of treating rectal varices, we chose the approach of double balloon-occluded embolotherapy (DBOE) with 5% ethanolamine oleate with iopamodol as a liquid embolic material. DBOE is one of the interventional radiology techniques (Morita et al., Acta Hepatol Jpn 1994;35:109-120), but in this case was a completely new and novel clinical procedure for rectal varices. After the DBOE therapy, the condition of rectal varices was markedly improved. Thus, DBOE might be a new tool for treating inoperable rectal varices.