A Simple Method for the Observation of Cultured Rat Hepatocytes by Scanning Electron Microscopy

A simple method for the three-dimensional observation of inner and outer ultrastructures of the cultured hepatocyte was presented. The freeze cracked plane of the cultured cell was obtained by means of cracking the underlying substrate. This method made it possible to observe the ultrastructural features of both cell surface and cytoplasm in the same cell. Among several substrates examined in this investigation, the collagen gel sheet was the most useful for this purpose

Morphological Observations of Rabbit Gastric Fundus Cells in Primary Culture

The morphological properties of the cells isolated from rabbit gastric fundus and the cells grown on a collagen gel sheet were investigated, using both light and scanning electron microscopes. The cells used for this study were obtained in the form of lumps from the isthmus region. The lumps consisted of 4 different types of cells (mucous and immatured cells, and a small number of chief and parietal cells). These isolated cells were then grow on collagen gel in a defined medium. However, two of those cells (chief and parietal cells) gradually disappeared as the culture time progressed. The majority of the cells grown on the gel were found to be mucous and assorted unidentified cells. The mucous cells were covered with many short microvilli, and many PAS positive granules in their cytoplasm. Some of those cells were covered with a mucous like substance. Judging from the staining properties of each cell, the unidentified cells appeared to indicate that they were immature cells. It was found that the cell's morphology and population retained constant regardless of whether Bh-EGF was administered or not

Isolation and functional characterization of a β-eudesmol synthase, a new sesquiterpene synthase from Zingiber zerumbet Smith

AbstractIn this paper, we have identified a new sesquiterpene synthase gene (ZSS2) from Zingiber zerumbet Smith. Functional expression of ZSS2 in Escherichia coli and in vitro enzyme assay showed that the encoded enzyme catalyzed the formation of β-eudesmol and five additional by-products. Quantitative RT-PCR analysis revealed that ZSS2 transcript accumulation in rhizomes has strong seasonal variations. To further confirm the enzyme activity of ZSS2 and to assess the potential for metabolic engineering of β-eudesmol production, we introduced a gene cluster encoding six enzymes of the mevalonate pathway into E. coli and coexpressed it with ZSS2. When supplemented with mevalonate, the engineered E. coli produced a similar sesquiterpene profile to that produced in the in vitro enzyme assay, and the yield of β-eudesmol reached 100mg/L

Effects of employment of distinct strategies to capture antibody on antibody delivery into cultured cells

The characteristics of antibody delivery into cultured HeLa cells were examined by using two delivery systems. Both systems used a cell-penetrating peptide as a tool for intrusion of an antibody into the cells, but either a “protein A derivative” or “hydrophobic motif” was employed to capture the antibody. When we examined the uptake of the Alexa Fluor-labeled antibody by use of these two systems, both systems were found to effectively deliver the antibody into the cultured cells. However, when we compared the amount of antibody delivered by these systems with the amount of transferrin uptake, the former was 10 times smaller than the latter. The lower efficiency of antibody delivery than transferrin uptake seemed to be attributable to the involvement of the antibody delivery reagent, which failed to catch the antibody molecule. This interpretation was validated by an experiment using a larger amount of antibody, and the amount of antibody delivered by the “protein A derivative” system under this condition was determined to be 13 ng proteins/105 cells. The antibody delivery achieved by the “protein A derivative” or “hydrophobic motif” showed two differences, i.e., a difference in intracellular distribution of the delivered antibody molecules and a difference in the fluorescence spectrum observed with cellular lysates. Possible reasons for these differences between the two delivery systems are discussed

ZnCl2 and vitamin C, known as antioxidants, differently potentiate the cytotoxicity of H2O2 in rat thymocytes : Cytometric analysis using forward and side scatters

The ‘antioxidant hypothesis’ proposes that antioxidant nutrients afford protection against chronic diseases by decreasing oxidative damages. The ability of zinc to retard oxidative processes has been recognized for many years. However, the application of ZnCl2 potentiates the cytotoxicity of H2O2. Thus, some antioxidants may be cytotoxic under certain oxidative conditions. Therefore, in this study, the effect of vitamin C, one of antioxidant nutrients, on the cells treated with H2O2 has been examined to see if vitamin C potentiates the cytotoxicity of H2O2. Experiments were carried out with flow cytometer and rat thymocytes. Vitamin C also potentiated the cytotoxicity of H2O2. The increase in cell lethality induced by the combination of H2O2 and ZnCl2 was associated with the increase in population of shrunken cells with increased intensity of side scatter. However, it was not the case for the combination of H2O2 and vitamin C. The profile of cytotoxicity induced by H2O2 and vitamin C was different from that by H2O2 and ZnCl2. It may be suggested that the effects of zinc and vitamin C varies from cytoprotective to cytotoxic, being dependent on the type of oxidative stress

Elastic Net Constraints for Shape Matching

We consider a parametrized relaxation of the widely adopted quadratic assignment problem (QAP) formulation for minimum distortion correspondence between deformable shapes. In order to control the accuracy/sparsity trade-off we introduce a weighting parameter on the combination of two existing relaxations, namely spectral and game-theoretic. This leads to the introduction of the elastic net penalty function into shape matching problems. In combination with an efficient algorithm to project onto the elastic net ball, we obtain an approach for deformable shape matching with controllable sparsity. Experiments on a standard benchmark confirm the effectiveness of the approach. © 2013 IEEE

Formation of High-Mass stars in an isolated environment in the Large Magellanic Cloud

The aim of this study is to characterize the distribution and basic properties of the natal gas associated with high-mass young stellar objects (YSOs) in isolated environments in the Large Magellanic Cloud (LMC). High-mass stars usually form in Giant Molecular Clouds (GMCs) as part of a young stellar cluster, but some OB stars are observed far from GMCs. By examining the spatial coincidence between the high-mass YSOs and 12CO (J = 1-0) emission detected by NANTEN and Mopra observations, we selected ten high-mass YSOs that are located away from any of the NANTEN clouds but are detected by the Mopra pointed observations. The ALMA observations revealed that a compact molecular cloud whose mass is a few thousand solar masses or smaller is associated with the high-mass YSOs, which indicates that these compact clouds are the sites of high-mass star formation. The high-density and high-temperature throughout the clouds are explained by the severe photodissociation of CO due to the lower metallicity than in the Galaxy. The star formation efficiency ranges from several to as high as ~ 40%, indicating efficient star formation in these environments. The enhanced turbulence may be a cause of the efficient star formation therein, as judged from the gas velocity information and the association with the lower density gas.Comment: 31 pages, 14 figures, PASJ accepted for publicatio

pH-resistant Inhibitor of Mitochondrial ADP/ATP Carrier

Bongkrekic acid (BKA), isolated from Burkholderia cocovenenans, is known to specifically inhibit the mitochondrial ADP/ATP carrier. However, the manner of its interaction with the carrier remains elusive. In the present study, we tested the inhibitory effects of 17 bongkrekic acid analogues, derived from the intermediates obtained during its total synthesis, on the mitochondrial ATP/ATP carrier. Rough screening of these chemicals, done by measuring their inhibitory effects on the mitochondrial ATP synthesis, revealed that 4 of them, KH-1, 7, 16, and 17, had moderate inhibitory effects. Further characterization of the actions of these 4 analogues on mitochondrial function showed that KH-16 had moderate; KH-1 and KH-17, weak; and KH-7, negligible side effects of both permeabilization of the mitochondrial inner membrane and inhibition of the electron transport, indicating that only KH-7 had a specific inhibitory effect on the mitochondrial ADP/ATP carrier. Although the parental bongkrekic acid showed a strong pH dependency of its action, the inhibitory effect of KH-7 was almost insensitive to the pH of the reaction medium, indicating the importance of the 3 carboxyl groups of BKA for its pH- dependent action. A direct inhibitory effect of KH-7 on the mitochondrial ADP/ATP carrier was also clearly demonstrated