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Volume 33 (2000)
- Issue 6 Pages 405-
- Issue 5 Pages 319-
- Issue 4 Pages 235-
- Issue 3 Pages 153-
- Issue 2 Pages 63-
- Issue 1 Pages 1-
Volume 33, Issue 5
Displaying 1-9 of 9 articles from this issue
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REVIEW
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Hiroshi Hasegawa, Vinci Mizuhira, Mitsuru Notoya2000 Volume 33 Issue 5 Pages 319-340
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSMore than a decade ago, we introduced a technique using a computer controlled household microwave oven for biological tissue fixation. This microwave irradiation fixation (MWIF) method has produced excellent results, enabling quick and homogeneous infiltration of tissues with a fixative containing chemical reagents. We also described the application of MWIF to light and electron microscopy, including those for enzyme and immunocytochemistry, autoradiography and the distribution of calcium ions. In this review, we present an overview of our experience. To ascertain that microwave irradiation (MWI) would enable aldehyde to quickly and homogeneously penetrate a tissue block, microautoradiography was performed using [3H]-formaldehyde as a tracer. With MWI, prominently developed silver grains were distributed homogeneously in the section, while the control section only had silver grains around it and not inside. Biochemical analysis revealed that complete tissue fixation occurred only after chemical cross-linking of proteins with the infiltrating aldehyde, which was a gradual process. This means that the tissue blocks must be left in the fixative for at least 30-60 min after MWI. To fix soluble peptides and protein or detect Ca2+ with MWI, we used a fixative containing a small amount of tannic acid (0.1%) or chemical reagents, which are very effective for these peptides and proteins, or Ca2+ in cells. Staining using MWI is also very fast and reliable for both light and electron microscopy.View full abstractDownload PDF (4987K)
REGULAR ARTICLES
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Rie Ikeda, Masanori Murakoshi, Masashi Ohtani, Masashi Tagawa, Takashi ...2000 Volume 33 Issue 5 Pages 341-347
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSTZT-1027, an inhibitor of tubulin polymerization, is a derivative of dolastatin 10 isolated from the Indian Ocean sea hare Dolabella auricularia. TZT-1027 has good anticancer activity. We investigated the effect on B16 melanoma of TZT-1027 in mice histopathologically and by agarose gel electrophoresis. At 1 mg/kg of TZT-1027, there was a good correlation between the number of mitosis-like cells and apoptotic bodies. In mitosis-like cells, the nuclear membrane had disappeared, and the chromosomes had morphological features typical of mitosis. However, not all cells in mitosis had proliferated. In contrast, at 2 mg/kg of TZT-1027, many apoptotic bodies were detected soon after treatment, but these were not involved in the dynamic changes to mitosis-like cells. Furthermore, TUNEL-positive cells were clearly visible at 1 hr after treatment and the nuclei of these cells showed chromatin margination, cap-shape chromatin clamping, and nuclear condensation on electron microscopy. Therefore, we speculated that the effect on B16 melanoma cells of TZT-1027 differed with the treatment dose, and a dose of 2 mg/kg induces apoptosis in tumor cells by a direct mechanism.View full abstractDownload PDF (1519K) -
Midori Ono2000 Volume 33 Issue 5 Pages 349-353
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSBecause angiogenesis is indispensable to the growth, invasion and metastasis of tumors, we conducted this study to elucidate the process of angiogenesis and the morphology of the new vessels in endometrial aspirated specimens. Cellular specimens from 21 cases of well differentiated endometrioid adenocarcinoma were employed in the investigation. Immunohistochemical staining was performed according to cell type using a CD34 monoclonal antibody as a vascular endothelial cell marker and α-smooth muscle actin as a pericyte marker. Microprojections that appeared to be sprouts of endothelial cells and finger-like processes that resembled endothelial cell extensions from the initial stage of angiogenesis were seen in a portion of the vascular network. Scattered cul-de-sacs that were in the process of extending into avascular fields were also observed. In some cases where several cul-de-sacs were observed, fork-like vessels were also seen, indicating that vigorous angiogenesis was occurring. This study elucidated the process of angiogenesis and the morphology of the new vessels in endometrial aspirated specimens.View full abstractDownload PDF (934K) -
Maria Luiza S. Mello, Silvya S. Maria Silvya S. Maria, Pedro H.P.A. Sc ...2000 Volume 33 Issue 5 Pages 355-359
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSThe frequency and responsiveness to the TUNEL assay were used to compare the DNA fragmentation associated with programmed cell death in erythrocytes of the circulating blood from chickens, pigeons, bullfrogs and tortoise species in which the life span and/or metabolic activities of these cells vary considerably. While most avian and adult bullfrog erythrocytes gave a positive response in this test, only 31% of the tortoise erythrocytes showed a positive TUNEL response, which was generally weak. The strongest positive response was seen in bullfrog erythrocytes. The positive results for avian erythrocytes may possibly be associated with their higher metabolic rate and body temperature while the lower response for tortoise erythrocytes may reflect the very long life span and lower metabolic rate of these cells. Based on previous literature reports, the results for the bullfrog are unlikely to be associated with the animal metabolic rate and do not necessarily indicate genomic inertness. Together with literature data showing divergent pathways for programmed cell death in erythrocytes, the differences in DNA fragmentation reported here indicate that further studies on vertebrate nucleate erythrocytes are required for a better understanding of cell death in these cells.View full abstractDownload PDF (810K) -
Toshiyuki Matsuzaki, Suzuki Takeshi, Shigeyasu Tanaka, Kuniaki Tanaka2000 Volume 33 Issue 5 Pages 361-365
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSRabbits were immunized with partial peptides corresponding to dog Na+-coupled myoinositol cotransporter (SMIT) and rat water channel aquaporin 3 (AQP3). Immunoblot analysis and immunofluorescence microscopy revealed that the antibodies corresponding to SMIT and to AQP3 both detected AQP3 protein in the kidney. Neither of them, on the other hand, detected SMIT protein. Immunogen peptides of both SMIT and AQP3 contain the identical sequence "KEQI", whereas other amino acids are different from each other. Immunohistochemical preabsorption studies demonstrated that the antibodies corresponding to SMIT mainly recognized the sequence "KEQI" as an epitope. These observations demonstrate that the utmost care should be taken in using anti-peptide antibodies.View full abstractDownload PDF (561K) -
Sheng-Chieh Lin, Steve R. Roffler, Tang-Lu Cheng, Kuo-Ting Chang, Jeff ...2000 Volume 33 Issue 5 Pages 367-375
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSChang hepatoma ascites (CHA) cells were previously used as an animal model for antibody-directed enzyme prodrug therapy (ADEPT) and internalization of surface lectin-binding sites in vivo and in vitro. A monoclonal antibody RH1 (Mab-RH1) that recognizes a CHA-specific antigen (CHAA) on the cell surface has been identified. Β -glucuronidase conjugated with Mab-RH1CHAA (Mab- β G-CHAA) inverts an antineoplastic prodrug, p-hydroxyaniline mustard glucuronide to toxic p-hydroxyaniline mustard and kills CHA cells. The possibility that CHAA may internalize into the tumor cells was hypothesized because the drug effect was reduced after long treatment. Nevertheless, the biological nature of CHAA and the mechanism of ADEPT remains unknown. In this study, various enzyme digestions characterize CHAA in situ. Furthermore, specific lectin-blocking methods determine the terminal monosaccharide that was recognized by Mab-RH1 on CHAA. The results indicate that the tumor-specific CHAA is a 32 kDa glycoprotein that is sensitive to protease treatment, but survives treatments of trypsin, chymotrypsin, lipase, neuraminidase, hyaluronidase, chondroitinase ABC and heparinase. Con A, WGA and RCA lectins block the recognition of Mab-RH1 and indicate that α-mannose, α-glucose, D-galactose and N-acetyl-D-glucosamine residues are essential for ADEPT. We conclude that internalization of Mab-βG-CHAA complexes causes the reduction of antineoplastic drug activity.View full abstractDownload PDF (1476K) -
Tetsuji Nagata2000 Volume 33 Issue 5 Pages 377-383
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSThe aging changes of the incorporations of radiosulfate (35SO4) into the tracheal cartilage were studied in several litters of mice at various ages, from embryos to postnatal 1 year by light microscopic radioautography. The results show that silver grains indicating the incorporations of radiosulfate were found over the cartilage matrices and the cartilage capsules in the hyaline cartilages of the tracheae of perinatal mice. The grain density was at the maximum at fetal day 19, then decreased from fetal day 19 to postnatal day 1, 3, 7, 14 and 30. The silver grains in the perinatal animals aged at postnatal day 1 and 3, moved from the internal layer to the external layer of the cartilage and from the interterritorial matrix to the territorial matrix and the cartilage capsule. No silver grains were found in the animals aged from 1 to 12 months. From these results, it is concluded that the radiosulfate was incorporated into sulfated complex carbohydrate in the cartilage matrices and the pattern of incorporation in the tracheae decreased with the aging of animals.View full abstractDownload PDF (1103K) -
Kazuharu Irie, Hidehiro Ozawa, Toshihiko Yajima2000 Volume 33 Issue 5 Pages 385-391
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSBased on their ultrastructural features, osteocytes have been classified into three phases: formative, resorptive, and degenerative. However, the mechanism of the morphological changes from formative osteocyte to resorptive osteocyte is still unclear. Therefore, we labeled bone matrix with calcein and examined chronologically the histochemical changes in osteocytes from two distinct areas in the mandible, focusing on the relationship between osteocytes and osteoclastic bone resorption. Tartrate-resistant acid phosphatase (TRAP) activity was detected only in osteocytes located near sites of osteoclastic bone resorption. Osteocytes far from osteoclasts lacked TRAP activity, even those that had been enclosed in bone matrix as long as the osteocytes that did show TRAP activity. Furthermore, glycogen granules evidenced by periodic acid-Schiff and the periodic acidthiocarbohydrazide-silver protein staining in osteocytes disappeared when the cell acquired TRAP activity. These findings suggest that the acquisition of TRAP activity in an osteocyte is closely synchronized with bone resorption. Disappearance of glycogen in osteocytes in the same area may also be related to the acquisition of TRAP activity or bone resorbing activity. The osteocytes characterized by glycogen accumulation must be in the phase that follows the formative phase, in which osteocytes display neither formation nor resorption activity.View full abstractDownload PDF (1623K) -
Hisashi Shiomi, Yasuyoshi Ohsaki, Akifumi Akamine, Tadahiko Iijima2000 Volume 33 Issue 5 Pages 393-398
Published: 2000
Released on J-STAGE: September 28, 2001
JOURNAL FREE ACCESSIt is well known that nerve growth factor (NGF) is expressed in the odontoblasts of developing teeth, but not in completely grown teeth in rats. However, there have been no detailed studies of the expression of NGF in reactivated primary odontoblasts or in newly differentiated odontoblast-like cells during pulp repair. Therefore, to examine the involvement of NGF in odontoblasts during pulp repair, we immunohistochemically observed the localization of NGF in dental pulp after cavity preparation in rat molars. Based on the depth of the cavity, two types of pulp reaction were thus distinguished. 1) In shal low cavities, primary odontoblasts secreted a new dentin matrix, reactionary dentin, and also expressed NGF. 2) In deep cavities, odontoblast-like cells, differentiated from pulpal mesenchymal cells, formed a large amount of reparative dentin or osteodentin. Additionally, intense NGF-like immunoreactivity was observed in odontoblast-like cells. These results suggest that NGF not only acts as a neurotrophic factor of dental pulp, but is also involved in the differentiation and functional change of odontoblasts during pulp repair.View full abstractDownload PDF (1095K)
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