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Biomedical subjects

H Asai

Publications and source records attributed to H Asai.

At least 91 records · Page 5Linked to original sources

Electron microscopic changes of bone marrow-derived cultured mast cells after injection into the skin of genetically mast cell-deficient W/Wv mice.

Phenotypes of bone marrow-derived cultured mast cells are different from those of connective tissue-type mast cells (CTMCs) that are found in the peritoneal cavity and the skin. When cultured mast cells of WBB6F1 - +/+ mouse origin were directly injected into the skin of genetically mast cell-deficient WBB6F1 - W/Wv mice, mast cells appeared in both the dermis and the subcutaneous tissue (beneath the panniculus carnosus). In contrast to cultured mast cells, mast cells that were observed in either the dermis or the subcutaneous tissue were stained with berberine sulfate, suggesting the content of heparin. Cultured mast cells acquired the electron microscopic features of CTMC in either the dermis or the subcutaneous tissue of WBB6F1 - W/Wv mice, but the electron density of mast-cell granules was significantly higher in the dermis than in the subcutaneous tissue. Such an electron microscopic difference was also observed after the injection of purified peritoneal mast cells of WBB6F1 - +/+ mice into the skin of WBB6F1 - W/Wv mice. From the present study, we suggest that the electron density of mast-cell granules in the skin of WBB6F1 - W/Wv mice is not dependent on the type of injected mast cells but on the anatomical sites at which the injected cells are located.

Animals↗

The relationship of SP and AP findings to hearing level in Menière's disease.

The relationship of summating potential (SP) and action potential (AP) to hearing threshold level was examined in 46 patients with a unilateral Ménière's disease. The SP and AP were recorded with the extratympanic method. The ratio of -SP and AP amplitudes in affected ears to those in contralateral ears (-SP and AP amplitude ratio) at 80 dB nHL of click stimuli was used as parameters of analysis in order to diminish the variability of SP and AP amplitudes among individuals. The results revealed that there is no relationship between -SP amplitude ratio and the hearing threshold level at any frequency, whereas AP amplitude ratio has a significant negative correlation (r = -0.419, p less than 0.01) to the average hearing threshold level at 2-8 kHz, but not at 0.25-1 kHz (p greater than 0.05). The results indicate that an increase in the ratio of -SP amplitude to AP amplitude with the deterioration of the hearing at higher frequencies reported by our previous study (Mori et al., 1987) results from a decrease in AP amplitude rather than an increase in -SP amplitude. The dependence of AP amplitude on hearing threshold level and the independence of -SP amplitude on hearing threshold level suggests that the mechanism underlying the increase in -SP amplitude may be different from that causing the hearing loss in Ménière's disease.

Acoustic Stimulation↗

Hybrid resistance to parental bone marrow-derived cultured mast cells in the skin but not in the peritoneal cavity of (WB X C57BL/6)F1-W/Wv mice.

When cultured mast cells of (WB X C57BL/6)F1-+/+(WBB6F1-+/+) and WB-+/+(WB) mice were directly injected into the skin of genetically mast cell-deficient WBB6F1-W/Wv mice, mast cell clusters appeared at the injection sites. Although in vitro colony-forming ability is comparable between cultured mast cells of WB mice and those of WBB6F1-+/+ mice, the number of WB mast cells necessary for the appearance of mast cell clusters in the skin of WBB6F1-W/Wv mice was significantly larger than the number of WBB6F1-+/+ mast cells. In spite of the presence of such an apparent hybrid resistance in the skin of WBB6F1-W/Wv mice to mast cells of the WB parent, both WB and WBB6F1-+/+ mast cells grow in the peritoneal cavity of WBB6F1-W/Wv mice with comparable efficiency. This is a demonstration of the tissue-related (nonrecirculating) expression of hybrid resistance against nonmalignant hematopoietic cells.

Animals↗

Changes in numbers and types of mast cell colony-forming cells in the peritoneal cavity of mice after injection of distilled water: evidence that mast cells suppress differentiation of bone marrow-derived precursors.

Two different types of cells in the peritoneal cavity of mice produce mast cell colonies in methylcellulose. "Large" mast cell colonies are produced by bone marrow-derived precursors resembling lymphoid cells by light microscopy (L-CFU-Mast), whereas "medium" and "small" mast cell colonies are produced by morphologically identifiable mast cells (M-CFU-Mast and S-CFU-Mast, respectively). In the present study we eradicated peritoneal mast cells by intraperitoneal (IP) injection of distilled water. The regeneration process was investigated to clarify the relationship between L-CFU-Mast, M-CFU-Mast, and S-CFU-Mast. After injection of distilled water, M-CFU-Mast and S-CFU-Mast disappeared, but L-CFU-Mast increased, and then M-CFU-Mast and S-CFU-Mast appeared, suggesting the presence of a hierarchic relationship. When purified peritoneal mast cells were injected two days after the water injection, the L-CFU-Mast did not increase. In the peritoneal cavity of WBB6F1-+/+ mice that had been lethally irradiated and rescued by bone marrow cells of C57BL/6-bgJ/bgJ (beige, Chédiak-Higashi syndrome) mice, L-CFU-Mast were of bgJ/bgJ type, but M-CFU-Mast and S-CFU-Mast were of +/+ type. The injection of distilled water to the radiation chimeras resulted in the development of bgJ/bgJ-type M-CFU-Mast and then S-CFU-Mast. The presence of mast cells appeared to suppress the recruitment of L-CFU-Mast from the bloodstream and to inhibit the differentiation of L-CFU-Mast to M-CFU-Mast.

Animals↗

Hand splint for rheumatoid arthritis patients during gait training after joint replacement in lower extremity.

Rheumatoid arthritis patients have multiple joint problems, often making it difficult to use gait aids after a joint replacement in the leg. To address this problem, we designed a hand splint with a hook on the palmar side for use with parallel bars. Patients put these splints on both hands and they can walk holding the bars with the hooks. Best suited for using this splint are rheumatoid arthritis patients who are unable to hold parallel bars without marked pain in the hands or fingers, contracture of the wrist joint, or dislocation of finger joints. Several patients have tried this splint, which worked safely and satisfactorily during exercise.

Arthritis, Rheumatoid↗

Multiple bidirectional alterations of phenotype and changes in proliferative potential during the in vitro and in vivo passage of clonal mast cell populations derived from mouse peritoneal mast cells.

Mouse peritoneal mast cells (PMC) express a connective tissue-type mast cell (CTMC) phenotype, including reactivity with the heparin-binding fluorescent dye berberine sulfate and incorporation of [35S] sulfate predominantly into heparin proteoglycans. When PMC purified to greater than 99% purity were cultured in methylcellulose with IL-3 and IL-4, approximately 25% of the PMC formed colonies, all of which contained both berberine sulfate-positive and berberine sulfate-negative mast cells. When these mast cells were transferred to suspension culture, they generated populations that were 100% berberine sulfate-negative, a characteristic similar to that of mucosal mast cells (MMC), and that synthesized predominantly chondroitin sulfate [35S] proteoglycans. When "MMC-like" cultured mast cells derived from WBB6F1-+/+ PMC were injected into the peritoneal cavities of mast cell-deficient WBB6F1-W/Wv mice, the adoptively transferred mast cell population became 100% berberine sulfate-positive. In methylcellulose culture, these "second generation PMC" formed clonal colonies containing both berberine sulfate-positive and berberine sulfate-negative cells, but exhibited significantly less proliferative ability than did normal +/+ PMC. Thus, clonal mast cell populations initially derived from single PMC exhibited multiple and bidirectional alterations between CTMC-like and MMC-like phenotypes. However, this process was associated with a progressive diminution of the mast cells' proliferative ability.

Animals↗

Phenotypic changes of bone marrow-derived mast cells after intraperitoneal transfer into W/Wv mice that are genetically deficient in mast cells.

The ability of mouse IL-3-dependent, bone marrow culture-derived mast cells (BMMC) to generate serosal mast cells (SMC) in vivo after adoptive transfer to mast cell-deficient mice has been defined by chemical and immunochemical criteria. BMMC differentiated and grown from WBB6F1-+/+ mouse progenitor cells in medium containing PWM/splenocyte-conditioned medium synthesized a approximately 350,000 Mr protease-resistant proteoglycan bearing approximately 55,000 Mr glycosaminoglycans, as defined by gel filtration of each. Approximately 85% of the glycosaminoglycans bound to the cell-associated BMMC proteoglycans were chondroitin sulfates based upon their susceptibility to chondroitinase ABC digestion; HPLC of the chondroitinase ABC-generated unsaturated disaccharides revealed these glycosaminoglycans to be chondroitin sulfate E. As determined by heparinase and nitrous acid degradations, approximately 10% of the glycosaminoglycans bound to BMMC proteoglycans were heparin. In contrast, mast cells recovered from the peritoneal cavity of congenitally mast cell-deficient WBB6F1-W/Wv mice 15 wk after intraperitoneal injection of BMMC synthesized approximately 650,000 Mr protease-resistant proteoglycans that contained approximately 80% heparin glycosaminoglycans of approximately 105,000 Mr. Thus, after adoptive transfer, the SMC of the previously mast cell-deficient mice were like those recovered from the normal WBB6F1-+/+ mice that were shown to synthesize approximately 600,000 Mr proteoglycans that contained approximately 80% heparin glycosaminoglycans of approximately 115,000 Mr. As assessed by indirect immunofluorescence staining and flow cytometry using the B1.1 rat mAb (an antibody that recognizes an epitope located on the neutral glycosphingolipid globopentaosylceramide), approximately 5% of BMMC bound the antibody detectably, whereas approximately 72% of the SMC that were harvested from mast cell-deficient mice 15 wk after adoptive transfer of BMMC were B1.1-positive; approximately 82% of SMC from WBB6F1-+/+ mice bound the antibody. These biochemical and immunochemical data are consistent with the results of previous adoptive transfer studies that characterized mast cells primarily on the basis of morphologic and histochemical criteria. Thus, IL-3-dependent BMMC developed in vitro, cells that resemble mucosal mast cells, can give rise in vivo to SMC that express phenotypic characteristics of connective tissue mast cells.

Animals↗

Changing processes from bone marrow-derived cultured mast cells to connective tissue-type mast cells in the peritoneal cavity of mast cell-deficient w/wv mice: association of proliferation arrest and differentiation.

Connective tissue-type mast cells (CTMC) and mast cells grown in vitro exhibit many differences in morphology, biochemistry, and function. When cultured mast cells of WBB6F1-+/+ mouse origin were injected into the peritoneal cavity of genetically mast cell-deficient WBB6F1-W/Wv mice, however, the cultured mast cells acquired characteristics similar to CTMC. In this study, we analyzed the changing process. When the density of the cultured mast cells was measured by Percoll density gradient centrifugation, the proportion of dense mast cells increased after injection into the peritoneal cavity. Because the increase in proportion of dense mast cells paralleled the increase in proportion of heparin-containing mast cells, both parameters may be used as an index for differentiation activity of cultured mast cells into CTMC. When proliferation activity of mast cells was estimated by the incorporation of bromodeoxyuridine, the proliferation activity decreased after the i.p. transfer. Moreover, when cultured mast cells were recovered 10 wk after the i.p. transfer, the mast cells almost lost proliferation activity in the same culture condition that had been used for establishment of cultured mast cells from the bone marrow of WBB6F1-+/+ mice. These results demonstrate that the proliferation arrest and the acquisition of CTMC-like characters are associated after i.p. transfer of cultured mast cells.

Animals↗

Mutual phenotypic changes between connective tissue type and mucosal mast cells.

Connective tissue mast cells (CTMC) and mucosal mast cells (MMC) are apparently different, but the ontologic relationship between these two types of mast cells was unclear. We investigated the relationship by combining in vivo and in vitro techniques. Although bone marrow-derived mast cells developed in suspension culture resemble MMC, they acquired histochemical, electron microscopical, biochemical and immunological characteristics of CTMC after the transfer into the peritoneal cavity of genetically mast cell-deficient WBB6F1-W/Wv mice. On the other hand, when single peritoneal mast cells (typical CTMC) of WBB6F1-+/+ mice were injected into the stomach wall of WBB6F1-W/Wv mice, CTMC appeared in the muscularis propria and MMC in the mucosa. These results suggest that CTMC and MMC may interchange and that their phenotypes are determined by the anatomical microenvironment in which the final differentiation occurs.

Animals↗

Long-term monoclonal reconstitution of erythropoiesis in genetically anemic W/Wv mice by injection of 5-fluorouracil-treated bone marrow cells of Pgk-1b/Pgk-1a mice.

The spleen colony-forming assay does not represent the number of hematopoietic stem cells with extensive self-maintaining capacity because five to 50 spleen colony-forming units (CFU-S) are necessary to rescue a genetically anemic (WB X C57BL/6)F1-W/Wv(WBB6F1-W/Wv) mouse. We investigated which is more important for the reconstitution of erythropoiesis, the transplantation of multiple CFU-S or that of a single stem cell with extensive self-maintaining potential. The electrophoretic pattern of hemoglobin was used as a marker of reconstitution and that of phosphoglycerate kinase (PGK), an X chromosome-linked enzyme, as a tool for estimating the number of stem cells. For this purpose, we developed the C57BL/6 congeneic strain with the Pgk-1a gene. Bone marrow cells were harvested after injection of 5-fluorouracil from C57BL/6-Pgk-1b/Pgk-1a female mice in which each stem cell had either A-type PGK or B-type PGK due to the random inactivation of one or two X chromosomes. When a relatively small number of bone marrow cells (ie, 10(3) or 3 X 10(3] were injected into 200-rad-irradiated WBB6F1-W/Wv mice, the hemoglobin pattern changed from the recipient type (Hbbd/Hbbs) to the donor type (Hbbs/Hbbs) in seven of 150 mice for at least 8 weeks. Erythrocytes of all these WBB6F1-W/Wv mice showed either A-type PGK alone or B-type PGK alone during the time of reconstitution, which suggests that a single stem cell with extensive self-maintaining potential may sustain the whole erythropoiesis of a mouse for at least 8 weeks.

Anemia↗

[A parallel comparative double blind study of cefixime with cefroxadine in the treatment of acute lacunar tonsillitis].

The clinical efficacy and safety of cefixime (CFIX), a new oral cephalosporin, were compared with those of cefroxadine (CXD) in patients suffering from acute lacunar tonsillitis in a double blind study. Two hundred and fifty two patients were given each orally 100 mg of CFIX b.i.d. or 250 mg of CXD t.i.d. for, in principle, 7 days. Number of patients evaluated for clinical efficacy was 202 (103 treated with CFIX and 99 treated with CXD). As for the backgrounds of patients, more severe cases were found in the CFIX group than in the CXD group (P less than 0.01). Efficacy rates evaluated by individual doctors were 88.3% in the CFIX group and 91.9% in the CXD group. There was no significant difference between the 2 groups. Efficacy rates on the third day after the initiation of treatment evaluated by the committee were 40.8% in the CFIX group and 47.9% in the CXD group with no significant difference. Efficacy rate on the 7th day, however, was 79.8% in the CFIX group and 93.4% in the CXD group, showing a significant difference (P less than 0.05). Bacteriological effectiveness were satisfactory for both groups with eradication rates of 93.4% for the CFIX and 96.9% for the CXD group. Number of patients evaluated for safety was 226 (110 treated with CFIX and 116 treated with CXD). No significant difference was observed between the 2 drug groups in incidences of side effects; gastrointestinal disturbances or rashes were noted in 6 patients (5.5%) of the CFIX group and in 5 patients (4.3%) of the CXD group. As for the abnormal laboratory findings, elevation of GOT & GPT was observed in 1 patients of the CFIX group. From these results, it was concluded that 100 mg b.i.d. of CFIX was as useful as 250 mg t.i.d. of CXD in the treatment of acute lacunar tonsillitis.

Acute Disease↗

Comparison of summating potential in Menière's disease between trans- and extratympanic electrocochleography.

The summating potential (SP) recorded concurrently with transtympanic (TT) and extratympanic (ET) electrocochleography in Menière ears was compared with respect to polarity and amplitude. There was a difference in SP polarity and amplitude between the two methods. The incidence of the increase in the ratio of -SP amplitude to the amplitude of the auditory nerve action potential (AP) (-SP/AP ratio) at 80 dB nHL as a response to a click was higher with the ET than with the TT method. Most of the ears exhibiting an increased +SP/AP ratio with the TT method showed an increased -SP/AP ratio with the ET method. The incidence of increased SP/AP ratios including -SP/AP and +SP/AP ratios was similar with both methods. A +SP was recorded in 14 of 25 ears upon stimulation with an 8-kHz tone burst with the TT method, whereas no +SP was recorded in 25 ears with the ET method. Tone bursts of 1 and 2 kHz produced a -SP and similar waveforms with both methods. Comparison of -SP amplitude recorded with both methods revealed that the ratio of -SP amplitude with both methods is different across the stimulus frequencies. The present study suggests that the -SP elicited by tone bursts of mid to low frequencies is more stable in the diagnosis of Menière's disease than -SP elicited by a click because of the lesser influence of the electrode location and that the increase in the +SP recorded with the TT method may be of clinical significance in the diagnosis of Menière's disease.

Action Potentials↗

Diagnostic value of extratympanic electrocochleography in Menière's disease.

In order to examine the usefulness of the noninvasive extratympanic electrocochleography (ECochG) in the diagnosis of Menière's disease, the results of extratympanic ECochG, performed on 28 normally hearing ears, 3 ears with otosclerosis and 325 ears with sensorineural hearing loss of various origins including 56 Menière ears, were analyzed. The characteristics of summating potential (SP) parameters in Menière ears were different from those of ears with hair cell loss due to streptomycin and kanamycin ototoxicity and noise, while there was no difference between the action potential (AP) of the cochlear nerve parameters of Menière ears and ears with hair cell loss. A much higher incidence of increased-SP/AP amplitude ratios in Menière's disease was observed than in hearing loss of other origins. The results demonstrate the usefulness of noninvasive extratympanic ECochG in the diagnosis of Menière's disease.

Action Potentials↗

[An electron microscopic study of malignant giant cell tumor of bone].

Three cases of malignant giant cell tumors were studied by light microscope and electron microscope. Light microscopic observation revealed three different types of tumor cells, namely, malignant stromal cells, fibrosarcomatous cell and osteosarcomatous cells. Electron microscopic observation revealed that fibrosarcomatous cells had ultrastructure similar to that of malignant stromal cells. Both types of cells showed nuclear pseudo-inclusions (NPI), and well developed organellae related to matrix synthesis in the cytoplasm. Polysaccharide staining of the same cells revealed positive products in the cytoplasm. Osteosarcomatous cells showed irregular nuclei which contained NPI and well developed r-ER. The cells were located in the non-calcified matrix which consisted of collagen fibers. No alkaline phosphatase activity was observed in this area. These facts suggest that: (1) malignant stromal cells and fibrosarcomatous cells synthesize collagenous stroma as normal fibroblasts do, and (2) osteosarcomatous cells are devoid of the ability to promote matrix calcification.

Adult↗