Search PubMed⌕ Search

Biomedical subjects

M Hiroi

Publications and source records attributed to M Hiroi.

At least 127 records · Page 7Linked to original sources
PubMed1993

Assessment of Chlamydia trachomatis-specific IgA and IgG serum antibodies in genitourinary Chlamydia trachomatis infection--comparative study between HITAZYME and IPAzyme.

We assessed the C. trachomatis antibody assay kit HITAZYME (Hitachi Chemical Co., Ltd.) using clinical specimens. This kit is based on an enzyme immunoassay (EIA) which utilized purified Chlamydia trachomatis outer membrane antigen as the solid phase antigen. Twenty-nine untreated male urethritis patients, 816 pregnant housewives, 188 cervicitis patients, and 76 pelvic inflammatory disease patients were tested. Agreement between the HITAZYME test and antigen detection in infected area was assessed, and a comparison was made with IPAzyme (a commercially available indirect immunoperoxidase assay kit). 1) Summary of HITAZYME and IPAzyme IgA: Agreement between the two assays was relatively good, i.e., 82.6% (916/1109). However, 5.5% (61/1109) were HITAZYME (-), IPAzyme (+), and 11.9% (132/1109) were HITAZYME (+), IPAzyme (-). Thus, in quite a few cases the results did not agree. IgG: Agreement between the two assays was 73.7% (817/1109). Agreement was relatively low, 24.4% (271/1109) were HITAZYME (-), IPAzyme (+). 2) In the cases of disagreement, more specific Western blot analysis was performed to check the reactivity of the anti-C. trachomatis antibody. When IgA was used, agreement between HITAZYME and Western blot analysis was 69.6% (16/23), and agreement between IPAzyme and Western blot analysis was 30.4% (7/23), whereas when IgG was used, agreement between HITAZYME and Western blot analysis was 80.0% (12/15), and agreement between IPAzyme and Western blot analysis was 20.0% (3/15). There was significantly greater agreement with HITAZYME than with IPAzyme. In other words, HITAZYME had greater specificity when reacted with C. trachomatis antigen than IPAzyme. 3) The IgA antibody-positive rate in antigen (+) cases (male urethritis: 72.7%, pregnant housewives: 65.7%, cervicitis: 70.3%, pelvic inflammatory disease: 70.0%) was significantly (p < 0.01) higher than in antigen (-) cases (male urethritis: 16.7%, pregnant housewives: 13.6%, cervicitis: 22.6%, pelvic inflammatory disease: 30.4%). Therefore, IgA antibody can serve as a suitable indicator for active infection. 4) The IgG antibody-positive rate in antigen (-) female cases was 15.5% using HITAZYME and significantly (p < 0.01) lower than with IPAzyme. HITAZYME had greater specificity than IPAzyme. In conclusion, HITAZYME has relatively good sensitivity and specificity. Moreover, since it is an EIA assay, it allows objective evaluation of results. It permits processing of a large number of specimens because it is easy to perform. Thus, HITAZYME is a superior antibody assay for C. trachomatis. It can be used when antigen tests are difficult to perform. It is strongly anticipated that HITAZYME will be able to be used clinically as a screening test.

Chlamydia Infections↗
PubMed1993

Aging changes in the alignment of chromosomes after human chorionic gonadotropin stimulation may be a possible cause of decreased fertility in mice.

The relation between the alignment of chromosomes in the aged oocyte and its capacities of fertilization and development of oocytes was examined. More embryos in the condition of insemination at 12 and 16 h after human chorionic gonadotropin (hCG) however grew into two and more cell stages than those in the insemination of 20, 24 and 28 h. Some embryos in the group of insemination at 12, 16 and 20 h after hCG injection had grown into the blastocyst stage. The embryos inseminated 24 h after hCG and later had shown no further development. Twelve hours after hCG injection, 67% of oocytes have chromosomes on a straight line (A), and 29% have chromosomes scattered in one group (B). Three percent of oocytes have them separated toward both spindle ends (C) and 2% of oocytes have no chromosomes (D). At 16 h after hCG, the alignments of chromosomes are almost the same as that of 12 h after hCG injection. At 20 h after hCG, the most common alignment of chromosomes was scattered in a small group. The alignment of chromosomes of most oocytes was splitting at 24 and 28 h after hCG injection. Thus the oocytes with the chromosome alignment of A or B can be fertilized and developed into the blastocyst stage. The alignment of A and B lasts only about 8 h after ovulation, and the time was extremely restricted for oocytes to develop normally.

Animals↗
PubMed1993

Effect of an aromatase inhibitor in human follicular fluid on DNA synthesis of granulosa cells, theca cells, and follicles.

An aromatase inhibitor was identified in human follicular fluid. We sought to examine the effects of this inhibitor on granulosa cells, theca cells, a mixture of granulosa and theca cells, and follicles. Granulosa cells, theca cells, the mixture of granulosa and theca cells, and follicles were incubated with [3H]-thymidine and aromatase inhibitor at 37 degrees C for 40 h in Ham F-10 medium. The radioactivities of acid-soluble fractions were counted. Aromatase inhibitor markedly inhibited the DNA synthesis of granulosa cells and follicles, but it did not inhibit the mixture of granulosa and theca cells. This suggested that aromatase inhibitor was a factor in the follicle paracrine system and acted mainly on granulosa cells.

Animals↗
PubMed1993

Role of the extracellular Ca2+ on the intracellular Ca2+ changes in fertilized and activated mouse oocytes.

Changes in intracellular calcium concentration ([Ca2+]i) in fertilized mouse oocytes were measured using the calcium-sensitive dye, fura-2. After fertilization, an initial long-lasting [Ca2+]i increase was followed by a periodic [Ca2+]i increase. The periodic increase in [Ca2+]i persisted for 1 to 3 h and all fertilized oocytes extruded the second polar body within 4 h. The mean interval of periodic [Ca2+]i increase was 470 +/- 180 s (mean +/- SD). The frequency of the periodic [Ca2+]i increase depended on the extracellular calcium concentration. Verapamil and nifedipine inhibited the periodic [Ca2+]i increase. Sixty-five per cent of tested cells extruded the second polar body within 90 min of exposure to 7% ethanol. In these activated oocytes, the long-lasting [Ca2+]i increase was observed. However, no cells showed a repetitive increase in [Ca2+]i. Both release of calcium from intracellular stores and influx of extracellular calcium contribute to the increase in [Ca2+]i induced by ethanol. We conclude that the extrusion of the second polar body requires an increase in [Ca2+]i above a certain threshold level and the mobilization of calcium of both the intracellular and extracellular space in mouse oocytes.

Animals↗
PubMed1993

Proliferating cell nuclear antigen (PCNA), immunostaining and flow cytometric DNA analysis of renal cell carcinoma.

Nuclear PCNA immunoreactivity is found in the proliferating compartment of normal and tumor cells. The purpose of the present study was to examine immunohistochemical staining of PCNA in renal cell carcinoma and its relationship to tumor grading, flow cytometric DNA analysis and prognosis. The resected kidneys from 81 patients with renal cell carcinoma were used. Anti-PCNA monoclonal antibody was applied to paraffin sections to identify proliferating cells by the SAB technique. The proportion of PCNA-positive cells to negative cells was determined by counting the tumor cells stained by PCNA at 400x magnification and expressed as percentage of positive cells. In parallel with the PCNA immunostaining DNA flow cytometry was performed, using the cell suspension prepared from paraffin sections and stained with propidium iodide. 79 tumors were evaluable for PCNA immunohistochemistry and 54 tumors for DNA flow cytometry. Immunohistochemically, there was no significant difference in percentage of PCNA-positive cells between tumor grades G 1 and G 2 (1.4 +/- 1.3% in G 1 and 1.6 +/- 1.4% in G 2), while G 3 tumors contained high percentage of PCNA positive cells, the mean value being 7.6 +/- 5.3%. The percentage of PCNA-positive tumor cells was found to be correlated with histological grading of tumor and also with the S+G2+M phase fraction and the S phase fraction measured by flow cytometry (p < 0.01). The mean percentage of PCNA-positive tumor cells was higher in aneuploid tumors than in diploid tumors. There was no significant relationship between the percentage of PCNA positive cells and tumor staging. Sixty-one cases were available for survival analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Aneuploidy↗
PubMed1993

Gap junction of stratum granulosum cells of mouse follicles: immunohistochemistry and electron microscopy.

The gap junction (GJ) of stratum granulosum cells was investigated in growing and mature follicles of the mouse by means of light- and electron-microscopic immunohistochemistry, freeze-fracturing and deep-etching methods. GJ increased in number and size in concert with the progress of follicular maturation and decreased rapidly in the preovulatory mature follicle. GJ membrane showed invagination, most remarkably at 5 h after hCG injection, within the cytoplasm of the granulosum cell. Two types of internalization seemed possible, annular internalization and highly convoluted tubular invagination, both by an actin-mediated mechanism. At least, a part of the internalized GJ membrane seemed to be degraded by a certain lysosomal enzyme (s). Connexin43 (derived from rat cardiac muscle GJ) was indicated to be a constituent of GJ protein in the mouse stratum granulosum cell. There was no GJ particle coexisting with tight junctional strands; by contrast, a formation plaque appeared quite often on granulosum cells at various stages. Such a plaque is presumed to represent a precursor structure of the new GJ of these cells. So far examined, however, the formation plaque failed to show positive reaction against anti-Cx43 antibody.

Animals↗
PubMed1992

Midline malignant B-cell lymphoma with leukemic transformation.

This article presents a case of lethal midline granuloma on the palate of a 44-year-old woman, which had been identified histologically as B-cell lymphoma with leukemic transformation in the terminal stages. At the first visit, physical and laboratory examinations showed no remarkable findings except for a necrotizing ulcer of the palate, and the biopsy specimens only showed massive inflammatory cell infiltration and necrosis of the granulation tissue. There was a short-term resolution after treatment with cyclophosphamide and prednisolone, but the disease reactivated and the necrotic ulcerative lesion progressively advanced into the nasal cavity. Specimens from the third biopsy exhibited histologic features that were consistent with malignant lymphoma of the diffuse, mixed B-cell type. Chemotherapy with the regimen of cyclophosphamide, doxorubicin, vincristine, and prednisolone was adopted but was interrupted after a short time because of bone marrow suppression. Subsequently, large numbers of enlarged abnormal lymphocytes with a few vacuoles in the cytoplasm appeared in the circulating blood, indicating leukemic transformation of the midline lymphoma. The patient died on the seventh day after the initiation of chemotherapy. In the presentation of this case, the authors mention clinically important matters regarding midline lethal lymphoma and briefly discuss the pathophysiology and pathogenesis.

Adult↗
PubMed1992

Factors promoting embryo implantation in in vitro fertilization and embryo transfer.

The low implantation rate after embryo transfer (ET) is the most important problem to be solved in in vitro fertilization and embryo transfer (IVF-ET). Various factors which may affect embryo implantation in the endometrium have been examined. In this study, the influence of the timing of luteinization on the establishment of pregnancy was investigated. Follicle stimulation was performed by luteinizing hormone-releasing hormone agonist, human menopausal gonadotropin and human chorionic gonadotropin (hCG). hCG was injected on the day when the mean diameter of two follicles exceeded 16 mm, and 36 h after the injection the oocyte was retrieved. Progesterone (P4) supplement was started on the day of ovum retrieval (50 mg), followed by 30 mg for 14 days. The patients were divided into three groups: (1) the pregnant patients (n = 20: group Pre); (2) the nonpregnant patients without bleeding during P4 injection (n = 35; group N), and (3) the nonpregnant patients with bleeding during P4 injection (n = 30; group B). The number of follicles that developed, endometrial thickness, the day of operation, the number of oocytes harvested, the number of good quality oocytes, the number of oocytes fertilized, the number of embryos transferred, estradiol (E2) and P4 levels, and the E2/P4 ratio were examined. The endometrial thickness of group Pre (10.9 +/- 0.6 mm; mean +/- SEM) on the day of hCG injection was significantly (p less than 0.05) greater than that of groups N (9.0 +/- 0.4 mm) and B (9.2 +/- 0.3 mm).(ABSTRACT TRUNCATED AT 250 WORDS)

Buserelin↗
PubMed1992

Decreased natural killer cell activity in women with endometriosis.

Natural killer (NK) cell activity is characterized by the spontaneous capacity of lymphoid cells derived from nonimmunized hosts to recognize and lyse certain tumor cell lines, virus-infected cells and transplanted tumor cell lines. Endometriosis is characterized by implantation and proliferation of autologous ectopic tissue in the pelvic cavity. Therefore, we focused on examining NK cell activity in women with endometriosis. NK cell activity in peripheral blood from women with endometriosis was lower than in women without endometriosis. On the basis of this finding, we analyzed also the effect of peripheral sera of women with endometriosis on NK cell activity. In the presence of peripheral sera of women with endometriosis, NK cell activity was significantly suppressed as compared with the sera of women without endometriosis. The suppressive effect of sera of women with endometriosis on NK cell activity showed dose-dependent curves. These studies provide the speculation that natural immunity mediated by NK cells may modulate the development of endometrial implants.

Cytotoxicity, Immunologic↗
PubMed1991

Immunoelectron microscopic identification of asialo GM1-positive cells in adult rat liver.

For the electron microscopic identification of asialo GM1-positive cells, fresh-frozen sections fixed with cold acetone and PLP-fixed vibratome sections of adult rat livers were prepared immunocytochemically using the avidin-biotin-peroxidase complex method. Asialo GM1-positive cells were located mainly in the sinusoids, and rarely in Glisson's sheath and portal veins. In the sinusoids, most pit cells, showing the ultrastructural characteristics of large granular lymphocytes (LGL), were positive for asialo GM1 but a few pit cells were asialo GM1-negative. There were several, morphological differences between asialo GM1-positive and -negative pit cells. The asialo GM1-negative pit cells were smaller and had less-developed cell organelles and fewer dense granules, suggesting a more immature stage of development. Almost all the monocytes, segmented neutrophils and eosinophils, and small or large lymphocytes in the sinusoids also showed positive reaction for asialo GM1. In Glisson's sheath, in addition to pit cells and lymphocytes, mast cells were also positive for asialo GM1. In contrast, fixed cells such as liver parenchymal cells, endothelial cells, Kupffer cells and Ito cells within the liver lobules, as well as biliary epithelial cells, smooth muscle cells, fibroblasts, endothelial cells and pericytes in Glisson's sheath were all negative for asialo GM1. Thus, cell surface asialo GM1 expression is not specific for pit cells (LGL) in the rat liver.

Animals↗
PubMed1991

Effects of clomiphene citrate on serum hormone levels and endometrial thickness in an in vitro fertilization and embryo transfer program.

Hormonal alterations and pregnancy rates were compared between the clomiphene citrate (CC)-hMG combination cycle and the hMG cycle alone in an in vitro fertilization and embryo transfer program. In the CC-hMG cycle, the numbers of follicles developed, oocytes recovered and embryos transferred were higher than those in the hMG cycle (p less than 0.01, p less than 0.01 and p less than 0.05, respectively). Pregnancy rates of the CC-hMG cycle (6.7%/operation, 7.1%/embryo transfer), however, were lower than those of the hMG cycle (25%/operation, 31.6%/embryo transfer). PRL levels in the hMG cycle increased according to estradiol levels. In the CC-hMG cycle, CC might inhibit estradiol-induced hyperprolactinemia and the increase was lower even if the estradiol level was much higher than that of the hMG cycle until day 2. On day 4, the PRL level in the CC-hMG cycle further increased. The thickness of endometrium on day -2 in the hMG cycle alone was much thicker than that in the CC-hMG cycle (9.8 +/- 0.4 vs. 6.8 +/- 0.4 mm, mean +/- SEM).

Clomiphene↗
PubMed1991

Embryonic heart rates: development in early first trimester and clinical evaluation.

One hundred and forty-three women in the early first trimester of gestation were examined 364 times using transvaginal sonography, and the development of embryonic heart rate was studied. In each case gestational age was revised retrospectively by either recorded basal body temperature or ultrasound crown-rump length dating between 9 and 10 weeks. Embryonic cardiac activity could be detected as early as 37 days of gestation. In 133 continuing pregnancies, embryonic heart rate rose from an average of 97.7 beats per min at 36-38 days to 174.7 beats per min at 60-62 days. A significant correlation was seen between gestational age and embryonic heart rate (p less than 0.001). The regression equation for heart rate was as follows: heart rate = 3.850 x gestational age (days) -54.64 (r = 0.908, n = 347), in short, embryonic heart rate continued to rise about 4 beats per min every day until 8 weeks of gestation. In this series, 10 pregnancies resulted in spontaneous abortion in the first trimester, and all of them showed relative bradycardia. Embryonic heart rate measurements in 8 of them were below the 95% prediction intervals for normal heart rate plotted against gestational age. This study suggests that embryonic heart rate measurement by ultrasound may be a new method for dating early first trimester, and that first trimester bradycardia may be associated with a poor prognosis for the pregnancy.

Abortion, Spontaneous↗
PubMed1991

Electron microscopic localization of acridine orange binding to euchromatin in human neuroblastoma cells.

The purpose of the present study was to examine the distribution pattern of acridine orange (AO) chromatin interaction products (AOCI) in human neuroblastoma IMR-32 cells and to test whether AO labeling is correlated with BrdU incorporation, and immunohistochemical localization of DNA polymerase alpha, and human N-myc-gene product. Effects of aphidicolin, alpha-amanitin, and actinomycin D on visualization of AO binding to euchromatin and on N-myc-gene expression were also examined. About 25% of the cell nuclei in logarithmic growth phase were immunohistochemically demonstrated to be labeled with BrdU after incubation at 37 degrees for 30 min, indicating cells in DNA synthesis. Most of the cell nuclei showed positive immunoreactivity to DNA polymerase alpha, while human N-myc gene product was found in about 60-80% of the cell nuclei. Electron microscopic studies revealed that about 25% of neuroblastoma cells showed characteristic AOCI within cell nuclei. In the presence of aphidicolin, alpha-amanitin, and actinomycin D, positive cells for N-myc gene product decreased markedly. Percentages of AO positive cells and numbers of AOCI per cell nucleus also showed a marked decrease. But northern blot analysis demonstrated that the expression level of N-myc gene was only repressed by the transcriptional inhibitors alpha-amanitin and actinomycin D. However, no repression was caused by aphidicolin. The present and previous studies of the authors suggest that the ultracytochemical AO method may be indicative for conformational changes of chromatin of cells confined to the cell cycle. Inhibitors of RNA and DNA synthesis then may change the conformational state of chromatin.(ABSTRACT TRUNCATED AT 250 WORDS)

Acridine Orange↗
PubMed1990

Distribution pattern of acridine orange chromatin interaction products in rat glioma C6 cells at different phases of the cell cycle.

The purpose of the present study was to examine the distribution pattern of electron-dense acridine orange (AO) chromatin interaction products in rat glioma C6 cells at different phases of the cell cycle. For synchronization in the early S-phase the cells in logarithmic growth were treated with 3 micrograms/ml aphidicolin, a specific inhibitor of DNA polymerase alpha and then cultured in normal medium. For synchronization in the M-phase the cells cultured with aphidicolin and then returned to normal medium were treated with 0.05 micrograms/ml colcemid. Histoautoradiographic analysis of the C6 cells using the pulse chase method demonstrated approximately 16 h of cell cycle time and about 6.5 h of S-phase. Ultracytochemically, AO chromatin interaction products were found in all phases of the cell cycle except for the mitotic phase, namely in G1, S, and G2. The highest percentage of AO chromatin interaction products was observed in the early S-phase and the lowest in the G2 phase. The mean number of AO chromatin interaction products per nuclear area increased in the course of S-phase parallel with an increase of 3H-uridine uptake during the S-phase. The results show a characteristic distribution pattern of AO label specific for each of the four stages of the cell cycle, however, the significance of the coincident RNA synthetic activity remains to be elucidated.

Acridine Orange↗
PubMed1990

Involvement of prolactin in the control of follicular maturation.

This study was designed to explore the role of prolactin (PRL) in the modulation of follicular growth and steroidogenesis. An ovarian perifusion technique was used to determine the effect of PRL on steroid production by ovarian tissue. Immature female rats were injected with 5 IU pregnant mare's serum gonadotropin to induce follicular maturation. After 40 h rats were killed, ovaries removed, and follicles dissected out. Follicles were divided into two groups (small, less than or equal to 4 mm; large, greater than or equal to 5 mm in diameter), cut into fine fragments, and perifused for 6 in the presence or absence of ovine PRL (100 or 1,000 ng/ml). In the group of small follicles, either of the PRL doses reduced the ability of follicular tissues to convert androgen to estrogen. HCG-induced production of progesterone by small follicles was also suppressed by higher doses of PRL. On the other hand, in the group of large follicles, PRL showed no effects on the aromatization and progesterone release by the follicular fragments. These results suggest that the small immature follicle is more sensitive to the inhibitory action of PRL on steroidogenesis and folliculogenesis. Thus it is possible that PRL may have a modulatory role in follicular growth and maturation and, consequently, in the selection of ovulatory follicle.

Androstenedione↗
PubMed1990

Turner's syndrome--review of the literature with reference to a successful pregnancy outcome.

A 30-year-old Japanese female with Turner's syndrome had become pregnant and delivered a normal male infant by cesarean section. Her menarche was at age 14, and secondary sex characteristics developed normally. She had some features of Turner's syndrome, and cytogenetic studies from peripheral blood lymphocytes and several tissues revealed only 45,XO karyotype without evidence of mosaicism. To our knowledge, she is only the thirteenth case of monosomy X to achieve pregnancy. A review of the literature indicates a high incidence of fetal wastage and increased chromosomal errors in the offspring of women with a 45,XO cell line with or without mosaicism.

Adult↗