Clonogenic Patterns of Human Pulmonary Adenocarcinoma Cell Lines (PC-9, PC-13 and PC-14) and How They Influence the Results of Test for Chemosensitivity to Cisplatin in the Human Tumor Clonogenic Assay

The clonogenic patterns of three human pulmonary adenocarcinomacell lines (PC-9, PC-13 and PC-l4) were studied by human tumorclonogenic assay (HTCA), and factors which could influence theresults of tests for the chemosensitivity of these tumor cellsto cisplatin in HTCA were determined. The results showed thateach tumor cell line had a characteristic clonogenic pattern.The time intervals for the cells to grow to the plateau phasevaried from 9 to 16 days, depending on the cell line and numberof cells plated. The number of cells plated could substantiallyinfluence the results of chemosensitivity tests. The percentageof surviving colonies increased markedly if too many cells (usually5 x 10⁴ or more/plate) were plated. For continuous exposure,the results of chemosensitivity tests were rather stable after7 days of incubation in each cell line, especially when fewerthan 2 x 10⁴ cells/plate were plated. For 1-hour exposure, theincubation periods for the results to become stable varied from7 to 16 days depending on the cell type, number of cells platedand drug concentration. It was stressed that for the correctevaluation of the chemosensitivity of cultured cell lines inHTCA, the clonogenic pattern of each tumor cell line shouldbe checked in detail before further experiments are conducted.The higher the concentration and the longer the exposure time,the more strongly cis-diamminedichloroplatinum (CDDP) suppressedthe colony growth in each of the three cell lines in HTCA, andit was recommended that CDDP should be used clinically in sucha way as to maintain a high serum level of the active form ofCDDP for a long time.     

Multiplex PCR-identified cutaneous tuberculosis evoked by Mycobacterium bovis BCG vaccination in a healthy baby

This is the first identified case of Mycobacterium bovis bacillus Calmette-Guerin (BCG)-derived cutaneous tuberculosis that localizes at a place different from the vaccination site in hosts without immune deficiency. A healthy baby with a developing abscess is described. A multiplex PCR identified the abscess as originating from M. bovis BCG Tokyo 172.     

Properties of an indirect composite material polymerized with two different laboratory polymerizing systems

The purpose of the current study was to evaluate the performance of two laboratory light polymerization systems used to polymerize an indirect composite (Sinfony). A two-step polymerization system (Visio-Alfa and Beta) and a halogen-metal halide unit (Twinkle MIII) were assessed. The composite was polymerized either with the Visio units or with the MIII unit for different exposure periods. Knoop hardness, water sorption, and solubility in water of the composite polymerized with the following modes were determined: Visio, 15 minutes; MIII, 30, 60, 90, 120, and 180 seconds. Extension of light exposure time to the MIII unit improved the hardness of the composite from 30.5 (30 s) to 40.7 (180 s), whereas hardness obtained with the Visio units resulted in 24.8 (15 minutes). Water sorption and solubility of the composite were greater when it was polymerized with the Visio units than with the MIII unit.     

A comparison of the effects of unfractionated heparin, dalteparin and danaparoid on vascular endothelial growth factor-induced tumour angiogenesis and heparanase activity

Disseminated intravascular coagulation (DIC) is the most common complication of solid tumours. In this study, the effectiveness of three polysaccharide anticoagulants (PSAs), at therapeutic doses, at inhibiting solid tumour growth was investigated. Mice with tumour xenografts were subcutaneously injected with either unfractionated heparin (UFH; 200 units kg(-1) day(-1)), dalteparin (75 units kg(-1) day(-1)) or danaparoid (50 units kg(-1) day(-1)). At these concentrations, these PSAs are equieffective at inhibiting blood coagulation activated factor X. In mice with Lewis lung carcinoma (LLC) tumours dalteparin and, to a lesser extent, UFH inhibited both tumour growth and angiogenesis, whereas danaparoid did not. In contrast, in mice with KLN205 tumours, all the PSAs inhibited tumour growth and angiogenesis. All the PSAs significantly inhibited proliferation, migration of endothelial cells and vessel formation in matrigel plugs containing vascular endothelial growth factor (VEGF) and there were no significant differences between these effects of the PSAs. The PSAs had no effect on endothelial cell tubular formation in vitro. Although all the PSAs inhibited VEGF production in KLN205 tumours in vivo and cells in vitro, in LLC tumours and cells only UFH and dalteparin inhibited VEGF production, whereas danaparoid did not. In both LLC and KLN205 tumours in vivo, heparanase activity was inhibited by UFH and dalteparin, but not by danaparoid. Hence, UFH and dalteparin may be more effective than danaparoid at inhibiting cancer progression in DIC patients with solid tumours, due at least in part to their ability to suppress VEGF and heparanase in tumours.     

Visualization of the Activity of Rac1 Small GTPase in a Cell

Rho family G proteins including Rac regulate a variety of cellular functions, such as morphology, motility, and gene expression. Here we developed a fluorescence resonance energy transfer-based analysis in which we could monitor the activity of Rac1. To detect fluorescence resonance energy transfer, yellow fluorescent protein fused Rac1 and cyan fluorescent protein fused Cdc42-Rac1-interaction-binding domain of Pak1 protein were used as intermolecular probes of FRET. The fluorophores were separated with linear unmixing method. The fluorescence resonance energy transfer efficiency was measured by acceptor photobleaching assisted assay. With these methods, the Rac1 activity was visualized in a cell. The present findings indicate that this approach is sensitive enough to achieve results similar to those from ratiometric fluorescence resonance energy transfer analysis.