巻号一覧
32 巻 (1999)
- 6 号 p. 449-
- 5 号 p. 381-
- 4 号 p. 299-
- 3 号 p. 203-
- 2 号 p. 103-
- 1 号 p. 5-
32 巻, 3 号
選択された号の論文の13件中1～13を表示しています
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Hayato Kawakami, Hiroshi Hirano1999 年 32 巻 3 号 p. 203-222
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーMouse colon carcinoma (colon 26) cells were shown to attach to cultured hepatic sinusoidal endothelial cells only in the presence of PRP (platelet-rich-plasma). Most tumor cells first attached to the endothelial cells, not directly but via aggregated platelets. In addition, almost all of the tumor cells that were attached to the endothelial cells showed positive staining with ECA (Erythrina crystagalli agglutinin), whereas nearly half of the cells before adhesion were negative for ECA binding. ECA-positive cells, when separated from ECA-negative ones, revealed strong potency for metastasis to the liver compared with ECA-negative ones. ECA-positive materials were extracted from the crude membrane fraction of colon 26 cells and were found to suppress the platelet adhesion. These results indicate that the ECA-positive sugar chains on the surface of colon 26 cells should mediate the binding between colon 26 cells and platelets and the attachment of tumor cells to the endothelium. The attachment of colon carcinoma cells to hepatic sinusoidal cells induced as above may cause the successive events in establishing cancer metastasis.抄録全体を表示PDF形式でダウンロード (8674K) -
Daisuke Aoki, Eiko Saitoh, Yumi Matsumoto, Eiichiro Tominaga, Akira Hi ...1999 年 32 巻 3 号 p. 209-214
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーEmploying RMG-1 cells derived from human ovarian adenocarcinoma and COS-1 cells transfected with cDNA for β1, 4-galactosyltransferase, we first investigated the intracellular localization of this enzyme and secondly assayed separately the two types of soluble forms of it (designated as normal GaIT and GAT) in their culture supernatants. As results, the binding of anti-β1, 4-galactosyltransferase monoclonal antibody was strongly positive at the Golgi area surrounding nuclei and the staining pattern was the same between RMG-1 cells and COS-1 transformants. These data demonstrated that the cells were producing β1, 4-galactosyltransferase and had the ability to release it into the culture medium. As for the two soluble forms released from the cells, they were simultaneously detected in the culture medium by Western blotting and enzyme immunoassay using monoclonal antibodies, i. e., Mab8628 reactive to both GAT and normal-GaIT and Mab8513 specific for GAT, indicating that both were derived from RMG-1 and from the cDNA introduced into COS-1 cells. Since serum-GAT clinically reflects tumor growth more accurately or specifically than normal GaIT does, RMG-1 and COS-1 cells having cDNA for β1, 4-galactosyltransferase would be promising as sources of these enzymes in order to investigate the differences in the behaviors of normal GaIT and GAT associated with ovarian malignant tumors.抄録全体を表示PDF形式でダウンロード (3710K) -
Tadashi Tai, Masaharu Kotani, Ikuo Kawashima1999 年 32 巻 3 号 p. 215-222
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーWe established an improved method for the generation of mouse monoclonal antibodies (MAbs) to glycosphingolipids especially gangliosides, sialic acid-containing glycosphingolipids, by immunizing mice with purified gangliosides. Using this method, we generated and characterized a large number of the MAbs specific for gangliosides. These MAbs enabled us to examine the distribution of gangliosides in the brain. Immunohistochemical studies revealed that (i) the expression of each ganglioside is restricted to subsets of neuronal cells and their surrounding neuropils in the central nervous system of the adult rat, including cerebrum, cerebellum, hippocampus, and spinal cord, and (ii) the expression of gangliosides changes during the development. Immunocytochemical studies also revealed a cell type-specific expression of gangliosides in primary cultures of the rat cerebellum. These results suggest that the specific gangliosides may play important roles in various phenomena, involving cell-cell recognition, neurite outgrowth, synaptogenesis, transmembrane signalling, apoptosis, and cell growth and differentiation in the central nervous system.抄録全体を表示PDF形式でダウンロード (5660K) -
Mitsunori Kokubo, Toshiro Higuchi, Ken-ichi Kudoh, Yoshimitsu Fukuda, ...1999 年 32 巻 3 号 p. 223-228
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーThe preparation of tissue sections for light microscopy is absolutely necessary in pathology. However, this process is still mainly performed manually. The preparation of many samples from a large number of specimens requires meticulous skills. In addition, it is very difficult to maintain a constant accuracy of the producedure.
To solve these problems we propose two new test machineries for automatic highspeed sample preparations. The first apparatus is conceptually based on attaching pressure-sensitive adhesive tape on the surface of a paraffin block prior to sectioning. This whole process could be repeated automatically. The sectioning is performed by a newly developed high-precision automatic microtome. Using the proposed machine, cut sections could be continuously mounted on the tape and stained automatically.
We also developed the second machine to mount sections on the object glass automatically by using static electricity. The principle of this method is based on electrostatically charging the surface of a paraffin block by which insulation tape was attracted and attached to the surface of the block. After sectioning the block, the section remains adhered on the insulation tape by electrostatic forces. Subsequently, the section is transferred to the object glass by using static electricity as well. In the use of such methods, tissue section on conventional object glass could be made fully automatic.抄録全体を表示PDF形式でダウンロード (6898K) -
Akihiko Kudo, Hiroshi Hirano1999 年 32 巻 3 号 p. 229-233
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーIn histochemistry, image analysis has become quite common and is now regarded as a powerful tool that gives us semi-quantitative and precise interpretation on the obtained image data. However, image analysis itself is a time-consuming and laborious work, and so researchers in the laboratory desire to make this task proceed as automatically as possible. In NIH Image and Adobe photoshop® software running on Power Macintosh computers, processes by automation are available in both programs, though they are quite different in each software. By use of both of these programs, the analysis processes become easier to set up. To automatically process the data, the suitable preprocessing of digital image data is required. That is, image processing programs are equipped with many processing programs, such as those for filtering, defining the threshold of images, or masking uninteresting areas. Additionally, a method for adequate labeling of the target substances and controlled conditions for the image digitization are always helpful.抄録全体を表示PDF形式でダウンロード (7031K) -
Kei-Ichi Hirai, Hiroki Shimada, Jiehong Pan, Seiichi Katoh, Eriko Sima ...1999 年 32 巻 3 号 p. 239-242
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーThe generation of reactive oxygen species in the mitochondria was investigated using electron microscopic cytochemistry and energy-filtering transmission electron microscopy in reference to the toxic mediator of paraquat poisoning. When isolated intact mitochondria from rat livers were reacted with paraquat in the presence of NADH, mitochondrial NADH-quinone oxidoreductase generated superoxide anions to cause the destruction of mitochondria which resulted in cell death. Cerium chloride captured hydrogen peroxide and formed cerium perhydroxide precipitates on the outer surface of the mitochondrial outer membrane. This localization was also clearly identified with electron spectroscopic imaging based on electron energy loss spectral analysis.抄録全体を表示PDF形式でダウンロード (2937K) -
Hideaki Tamaki, Shohei Yamashina1999 年 32 巻 3 号 p. 243-252
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーTo correlate the structure and function of the Golgi apparatus, we employed computergenerated three-dimensional reconstruction from electron micrographs of serial sections treated for thiamine pyrophosphatase (TPPase) cytochemistry. Three-dimensional analyses were conducted for rat parotid acinar cells in a resting state, in the course of mitotic division, and during postnatal development.
In resting acinar cells, the TPPase-positive trans elements are the same size as the TPPase-negative middle/cis elements horizontally, and they mutually overlap from side to side. During the mitotic stage, TPPase activity disappears from the disorganized Golgi apparatus and no secretory granule formation is observed. Recovery of the stacked structure and TPPase activity take place in daughter cells at the telophase, which are followed by secretory granule formation. During the differentiation of Golgi apparatus in postnatally developing acinar cells, TPPase activity is expressed following formation of the stacked structure, and then granule formation is initiated. These results clearly demonstrate the close relationship among the stacked structure, TPPase activity and the exocrine function of the Golgi apparatus.抄録全体を表示PDF形式でダウンロード (8480K) -
Takehiko Koji1999 年 32 巻 3 号 p. 255-260
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーTo understand better the regulation of gene expression, an analysis of the expression of trans-acting transcription regulatory factors in individual cells is essential. Although cellular analysis of specific mRNA expression is successfully performed by in situ hybridization, the results are not enough to tell whether an elevated level of certain mRNA is due to an increased rate of transcription of the gene or due to a decreased rate of degradation of the mRNA. Recently, we have developed a new method, Southwestern histochemistry, which allows us to localize transcription regulatory factors in situ. In principle, double-stranded (ds) DNA harboring specific response element base sequences is synthesized and labeled with a hapten such as thymine-thymine (T-T) dimer and digoxigenin (Dig). After the reaction of frozen tissue sections with the probe ds DNA, the signal is detected enzyme-immunohistochemically by horseradish peroxidase (HRP) -labeled anti-T-T dimer or anti-Dig, respectively. As a model system, we localized estrogen receptor (ER) in mouse uterine tissue sections by immunohistochemistry with 1D5 monoclonal antibody and Southwestern histochemistry using T-T dimerized estrogen responsive element (ERE) ds DNA of vitellogenin. Consequently, very similar localization of ER was obtained by both methods. This result, together with other examples, further indicates the usefulness of Southwestern histochemistry as a method to localize specific transcription regulatory factors in situ.抄録全体を表示PDF形式でダウンロード (4326K) -
Yoichi Tani1999 年 32 巻 3 号 p. 261-270
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーThe method of in situ hybridization (ISH) using non-radioactive detection systems has become a very important molecular tool in research and diagnosis. However, nonradioactive ISH techniques generally detect only relatively abundant DNA or RNA molecules. The methods of target amplification, which multiply target molecules, or the methods of signal amplification, which increase the sensitivity of visualization, have been introduced for the detection of rare targets. Polymerase chain reaction (PCR) has been employed in most approaches to target amplification. Although the general principle of PCR in situ amplification is simple, its practical methods have revealed poor amplification efficiency and reproducibility as well as target localization. We have evaluated PCR in situ amplification methods to detect single copies of viral DNA with an experimental model of cervical carcinoma cell lines, and compared them with catalyzed signal amplification methods that we have developed using hapten-conjugated tyramide. Both amplification methods demonstrated the capability of detecting a single copy of DNA. In this overview, the applications and limitations of both target and signal amplification are summarized.抄録全体を表示PDF形式でダウンロード (9159K) -
Haruo Ohtani1999 年 32 巻 3 号 p. 271-274
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーDetection of tissue mRNA in human surgical materials by non-radioactive in situ hybridization technique may be difficult owing to inconsistent retention of tissue mRNA. This inconsistency is generally ascribed to different timing of ligation of feeding arteries during surgical operation. To improve this inconsistency, we adopted fixation in 4% paraformaldehyde/0.5% glutaraldehyde for 12-16hr with paraffin embedding. This method resulted in improved retention of tissue mRNA: higher in sensitivity and more consistent with cases. Furthermore, this procedure has shortened the color development time of the labeled enzyme, alkaline phosphatase. This helps to reduce nonspecific reactivity because prolonged color development time (up to 24) is one of the major causes of non-specific reactions. Cut sections can be stored at 4°C for more than a year, not resulting in detectable reduction of sensitivity. These are major advantages of this method particularly as human surgical specimens are concerned, because accumulation of cases is important for clinicopathologic analyses. Therefore, this fixation method is, at least, recommendable to improve in situ hybridization technique.抄録全体を表示PDF形式でダウンロード (2727K) -
Katsumasa Takahashi, Ken Kitamura, Toshihiro Takizawa1999 年 32 巻 3 号 p. 275-280
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーTo improve the sensitivity of in situ hybridization (ISH) carried out using oligonucleotide probes without radionuclides, we employed a new method entailing amplification of ISH signals by catalyzed reporter deposition. This method is based on biotin-tyramine-catalyzed signal amplification (BT-CSA) and was used to detect expression of a relatively scarce polynucleotide, kinesin 3A mRNA, in the central nervous system. Strong signals were detected in the hippocampus and in the granular cell layer of the cerebellum, but expression of kinesin 3A mRNA was not detected in Purkinje cells. ISH performed without the use of radionuclides has the advantage of being comparatively easy to perform, but has the disadvantage of low sensitivity. We showed that the low sensitivity can be overcome by using BT-CSA. Our method also had the advantage of enabling visualization of ISH signals via enzymatic reaction with horseradish peroxidase or by fluorescent staining.抄録全体を表示PDF形式でダウンロード (3799K) -
Huagang Zhu, Yoichiro Kato, Reiko Tanaka, Takao Obara, Kanji Sato, Mak ...1999 年 32 巻 3 号 p. 281-285
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーOsteopontin (OPN) is a secreted, adhesive glycophosphoprotein that has been implicated in both normal and pathologic processes. In recent years, it has been suggested that OPN expression also plays an important role in tumorgenesis, tumor progression and tumor metastasis. To investigate OPN expression in papillary thyroid carcinoma, thirty paraffin sections of papillary thyroid carcinoma confirmed pathologically were examined by immunohistochemistry and in situ hybridization. Immunopositive reactions for OPN-protein were obtained in the nuclear and perinuclear area in all cases. OPN mRNA was expressed in the carcinoma cells around the microcalcifications. The results demonstrated that OPN mRNA is expressed not only by the infiltrated macrophages but also by the papillary thyroid cancer cells. Hence, OPN expression can be considered a pathological feature of papillary thyroid carcinoma and further study into the relation between OPN and the biological behavior of papillary thyroid carcinoma is warranted.抄録全体を表示PDF形式でダウンロード (1459K) -
Young-Sik Kim, Sang Ju Lee, Insun Kim, Gyoung-Mi Kim, Seung R. Paik1999 年 32 巻 3 号 p. 287-294
発行日: 1999年
公開日: 2010/02/26
ジャーナル フリーThis study was carried out to show whether the removal of tissue-bound calcium ions during a heat-induced epitope retrieval (HIER) procedure improved immunohistochemical staining of the tissues. In addition, the proteolytic susceptibilities of the fixed tissues before and after various epitope retrieval procedures were also evaluated. Immuno-histochemical staining for MIB-1 antigen from paraffin-embedded cases of tonsillitis, gastric adenocarcinomas, and breast carcinomas was performed following heat-treatments for 1, 2, 3, and 5 min in the presence of various buffers such as 0.01 M citrate buffer (pH 6.0), 0.1 M Tris-HCI buffer (pH 8.0), and 1 mM EDTA-NaOH solution (pH 8.0), each of which has different calcium chelating capabilities. In addition, the immuno-histochemical staining was performed following 0.05% and 0.1% trypsinizations of the tissues before and after the epitope retrieval. The trypsin-treated tissues were also analyzed with sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
The immunohistochemical staining of MIB-1 was most prominent when 1 mM EDTA solution (pH 8.0) was employed during HIER. Although trypsin digestion of the tissues before and after HIER generally increased the staining intensity, the cellular morphology of the background was poorly preserved. In addition, no significant differences were observed with gel electrophoresis of the trypsin digested tissues prepared in the various epitope retrieval solutions. Taken together, it can be suggested that the EDTA solution is more effective than other buffers in exposing epitopes presumably by removing tissue-bound calcium ions. Also, an additional trypsin treatment is not necessary even before and after the HIER as long as the EDTA solution is used.抄録全体を表示PDF形式でダウンロード (8475K)
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