Molecular Mechanisms and Tumor Biological Aspects of 5-Fluorouracil Resistance in HCT116 Human Colorectal Cancer Cells

5-Fluorouracil (5-FU) is a cornerstone drug used in the treatment of colorectal cancer (CRC). However, the development of resistance to 5-FU and its analogs remain an unsolved problem in CRC treatment. In this study, we investigated the molecular mechanisms and tumor biological aspects of 5-FU resistance in CRC HCT116 cells. We established an acquired 5-FU-resistant cell line, HCT116R^F10. HCT116R^F10 cells were cross-resistant to the 5-FU analog, fluorodeoxyuridine. In contrast, HCT116R^F10 cells were collaterally sensitive to SN-38 and CDDP compared with the parental HCT16 cells. Whole-exome sequencing revealed that a cluster of genes associated with the 5-FU metabolic pathway were not significantly mutated in HCT116 or HCT116R^F10 cells. Interestingly, HCT116R^F10 cells were regulated by the function of thymidylate synthase (TS), a 5-FU active metabolite 5-fluorodeoxyuridine monophosphate (FdUMP) inhibiting enzyme. Half of the TS was in an active form, whereas the other half was in an inactive form. This finding indicates that 5-FU-resistant cells exhibited increased TS expression, and the TS enzyme is used to trap FdUMP, resulting in resistance to 5-FU and its analogs.     

Physico-chemical properties of glycyrrhizic acid in aqueous media. I. Surface-active properties and formation of molecular aggregates (author''s transl)

As shown in 138 embryos and fetuses in the 40-285 mm crown-rump length range, carpal and carpal-metacarpal "fusions" arise from incomplete separation of the cartilaginous precursors rather than from failure of initiation, thus accounting for the "fusions" seen in postnatal radiographs and the grooves that are evident enough in adult fusions. Radiographs selected from over 20,000 apparently normal individuals provide postnatal counterparts for the prenatal examples shown in histological sections.     

The effect of roll-processing orientation on stress corrosion cracking of warm-rolled 304L stainless steel in oxygenated and deoxygenated high temperature pure water

Stress corrosion crack growth rates of 304L stainless steel after one-directionally warm-rolled along the longitudinal (L) direction were measured in oxygenated and deoxygenated pure water at 288 °C. The fracture mode is mainly intergranular in the Transverse-longitudinal (T-L) orientation specimen, while it is occasionally intergranular in the Longitudinal-transverse (L-T) orientation specimen. The crack growth rates in the T-L orientation are higher than those in the L-T orientation in both oxygenated and deoxygenated environments, which is related to the effective cracking growth path as the result of microstructural anisotropy and applied loading direction.     

A subcloned human esophageal squamous cell carcinoma cell line with low thrombomodulin expression showed increased invasiveness compared with a high thrombomodulin-expressing clone--thrombomodulin as a possible candidate for an adhesion molecule of squamous cell carcinoma

Inside-out vesicles of plasma membranes prepared from a plant source were used as models to investigate effects of centrifugal forces on separations of early and late endosome populations by aqueous two-phase partition. Endosome subpopulations were resolved readily by preparative free-flow electrophoresis where acidification of the interiors of late endosomes occurred upon addition of ATP to activate a proton translocating ATPase. The resultant increased diffusion potential provided for a surface difference between late and early endosomes to permit electrophoretic separation. With the plant membranes, unincubated inside-out plasma membrane vesicles modeled early endosomes, whereas inside-out vesicles incubated with 1 mM ATP modeled late endosomes. A latent, 2,4-dichlorophenoxyacetic acid (2,4-D)-(auxin)-stimulated NADH:protein disulfide reductase measured spectrophotometrically was used as an enzymatic marker for both populations of inside-out vesicles. Phase partition behavior of each population was quantitated using total protein as the parameter.     

Cytochemistry of gastric parietal cells with high-pressure freezing followed by freeze-substitution

Gastric parietal cells were examined for changes in their ultrastructure and distribution of the proton pump during feeding and fasting states in rats. The fundic glands from rats fed ad libitum or fasted with free access to water were cryofixed using high-pressure freezing followed by freeze-substitution in acetone containing osmium or acrolein and then embedded in Epon 812 or Lowicryl K4M resin, respectively. Excellent ultrastructural preservation was achieved. During the feeding state, intracellular canaliculi and numerous microvilli were well developed, while tubulovesicles were poorly developed. In contrast, during the fasting state, the microvilli in the narrowed space of the intracellular canaliculi were tightly packed and the tubulovesicles were enlarged. Ultrathin sections were immunostained with antibodies against the alpha- and beta-subunits of the proton pump, H+ x K(+)-ATPase, using the immunogold method. The labelling was strong and clearly localized in comparison with that obtained using the conventional chemical-fixation method. Each subunit was localized on the membrane of the microvilli, intracellular canaliculi and tubulovesicles. The distribution of subunit proteins varied between the two states. During ad libitum feeding, the immunolabelling was localized strongly on the membranes of the microvilli and intracellular canaliculi. In contrast, the labelling was strong on the tubulovesicle membrane in the fasting state. The results obtained with each anti-subunit antibody by H+ x K(+)-ATPase immunostaining revealed differences in distribution and labelling density between the feeding and fasting states.     

Formation of large DNA aggregates induced by spermidine

The formation of large DNA aggregates induced by spermidine was investigated by UV absorptiometry and polarizing microscopy. The present results reveal that it is stepwise and involves the following morphological variations: fiber, fiber bundles, and a highly condensed phase. Furthermore, the influence of DNA concentration on not only the spermidine concentration required for the DNA aggregation but also the concentration of free spermidine during the aggregation is analyzed.     

Decorrelation of SAR data by urban damages caused by the 1995 Hyogoken-nanbu earthquake

Large part of urban area of around Kobe city were damaged by the 1995 Hyogoken-nanbu earthquake. For detecting the damaged area, the authors computed correlation coefficients from single-look complex ERS-1/SAR data. Two types of correlations, intensity correlation and coherence were computed. Decorrelation was found in the damaged urban built-up area in the data pair of which interval included the earthquake occurrence. The similarity of the decorrelation between the intensity correlation and the coherence indicated that major factor of the decorrelation is closely related to interferometric processes, which results in the change of speckle patterns in the single-look intensity images. A normalized difference was calculated from the correlation coefficients between the data pair including the earthquake occurrence and the pair before the earthquake. The distribution patterns of the pixels for which normalized difference was higher than a threshold showed good correspondence with the result of the ground survey. The result of this study indicates the possibility of detecting urban disasters using the decorrelation of SAR data.