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Biomedical subjects

M Hiroi

Publications and source records attributed to M Hiroi.

At least 91 records · Page 5Linked to original sources

Ovarian fecundity in patients with endometriosis can be estimated by the incidence of apoptotic bodies.

OBJECTIVE: To investigate the incidence of apoptotic bodies in membrana granulosa in patients with endometriosis. DESIGN: Retrospective analysis. SETTING: Yamagata University Hospital. PATIENT(S): Fifty-eight normoovulatory women with endometriosis and 45 patients with male factor infertility underwent ovulation induction for IVF-ET with GnRH analogues and gonadotropins. INTERVENTION(S): Patients underwent follicle aspiration after the administration of hCG. MAIN OUTCOME MEASURE(S): Recovered granulosa cells (GCs) were examined by fluorescence microscopy, and the incidence of apoptotic bodies was tabulated. These data and the other parameters associated with IVF results were analyzed on the basis of both the revised American Fertility Society (AFS) classification of endometriosis and the existence of chocolate cysts. RESULT(S): The incidence of apoptotic bodies in membrana granulosa of patients with endometriosis was significantly higher than that of the control (male factor infertility) group and increased as the stage of the revised AFS classification advanced. The incidence of apoptotic bodies in membrana granulosa was significantly higher in patients with chocolate cysts than in those without chocolate cysts. The patients with endometriosis had smaller numbers of developed follicles (> or = 15 mm), harvested oocytes, and mature oocytes than the male factor infertility patients. The existence of chocolate cysts corresponded with a reduced number of both harvested oocytes and mature oocytes. CONCLUSION(S): Through the apoptosis of GCs, the existence of endometriosis may have negatively affected the follicle development and oocyte quality.

Adult↗

Characterization of cationic amino acid transporter and its gene expression in rat hepatic stellate cells in relation to nitric oxide production.

BACKGROUND/AIMS: Nitric oxide is a potent mediator of hepatic sinusoidal hemodynamics and affects hepatic stellate cells (Ito cells, fat-storing cells). Although nitric oxide production may depend on the induction of inducible nitric oxide synthase and on transport of extracellular L-arginine, the precise mechanisms controlling nitric oxide production in stellate cells have not been well characterized. METHODS: Using stellate cells prepared from the male Wistar rat, kinetic analysis of L-arginine transport and reverse transcription-polymerase chain reaction for cationic amino acid transporter were carried out. The effect of tumor necrosis factor-alpha and interferon-gamma on L-arginine transport, mRNA expression of cationic amino acid transporter and inducible nitric oxide synthase, and nitric oxide production of stellate cells was assessed. RESULTS: The L-arginine transport system functioning in the transformed hepatic stellate cells was system y+, possibly mediated by cationic amino acid transporter-1 and cationic amino acid transporter-2B (Km approximately 50 microM). Tumor necrosis factor-alpha enhanced cationic amino acid transporter-2B mRNA expression and L-arginine transport, whereas cationic amino acid transporter-1 mRNA expression remained unchanged. Interferon-gamma induced the expression of inducible nitric oxide synthase mRNA without obvious changes in L-arginine transport. Interferon-gamma in combination with tumor necrosis factor-alpha induced nitric oxide production with an enhancement in cationic amino acid transporter-2B mRNA expression, inducible nitric oxide synthase mRNA expression, and L-arginine transport, while extracellular L-lysine competitively inhibited this nitric oxide production. CONCLUSIONS: In transformed hepatic stellate cells, tumor necrosis factor-alpha and interferon-gamma have a crucial role in nitric oxide production, and extracellular L-arginine transport and inducible nitric oxide synthase expression are regulated in a differential cytokine-specific manner. As the estimated Km of L-arginine transporter in transformed hepatic stellate cells is very similar to the physiological L-arginine concentration in portal vein, we assume that increased portal L-arginine concentration may easily affect sinusoidal blood flow through enhancement of autocrine nitric oxide production in transformed hepatic stellate cells of diseased liver.

Amino Acid Transport Systems, Basic↗

The role of myofibroblasts at the tumor border of invasive colorectal adenocarcinomas.

BACKGROUND: In order to elucidate the significance of myofibroblasts in invasive growth of colorectal adenocarcinomas, we examined the number of myofibroblasts at the tumor border of colorectal adenocarcinomas. METHOD: A total of 91 invasive colorectal adenocarcinomas were examined immunohistochemically using anti-alpha-smooth muscle actin (ASMA) and high-molecular-weight caldesmon (h-CD) antibodies; 25 carcinomas confined to the submucosa (sm carcinomas), 40 carcinomas confined to the muscularis propria (mp carcinomas) and 26 carcinomas invading the subserosa or adventitia (ss carcinomas). We considered ASMA-positive and h-CD-negative stromal cells as myofibroblasts. RESULTS: Twenty-seven (67%) of the 40 mp carcinomas and 25 (96%) of the 26 ss carcinomas had a small number of myofibroblasts at the tumor border facing the muscularis propria. CONCLUSIONS: Although direct evidence is lacking, there is a possibility that the further immediately vertical and radial invasion of carcinoma cells into the subserosa or adventitia is associated with a smaller number of myofibroblasts at the tumor border facing the muscularis propria in mp carcinomas, resulting in a low incidence of mp and a high incidence of ss carcinomas in the colorectum.

Adenocarcinoma↗

Well differentiated adult-type fibrosarcoma arising from the occipital subcutaneous tissue in a 17-year-old man: case report with immunohistochemical study.

We report a case of a 17-year-old man with a spindle cell tumor in the occipital subcutaneous tissue. The enucleated tumor, measuring 2.5 x 2.0 x 1.0 cm, had a broad-bean shape and well circumscribed border and was localized to the subcutis without dermal involvement. Microscopically, the tumor was composed of uniform spindle cells showing interlacing bundle formation and a herringbone pattern. The neoplastic cells were separated by collagen fibers in parallel fashion, the amount of which varied with different areas in the tumor. Mitotic figures were eight mitoses per 50 high-power fields in number. The neoplastic cells were positive for vimentin, Factor-XIIIa, alpha-smooth muscle actin and CD34, but negative for desmin, calponin, high molecular weight caldesmon, smooth muscle myosin heavy chain, collagen type IV, laminin and S-100 protein. These immunohistochemical results indicated that the neoplastic cells showed differentiation toward fibroblasts/myofibroblasts/dendritic interstitial cells. Although more than 50% of the neoplastic cells were positive for CD34, the present tumor should be diagnosed as well differentiated fibrosarcoma, adult type, rather than extrapleural solitary fibrous tumor and fibrosarcomatous areas of dermatofibrosarcoma protuberans, on the basis of routine microscopic findings mentioned above.

Adolescent↗

Partial characterization of the gametes and development of a successful in vitro fertilization procedure in the mastomys (Praomys coucha): a new species for reproductive biology research.

The mastomys, a small African rodent, has been extensively used as a laboratory animal for cancer research. Previous preliminary studies have demonstrated that the animal has a relatively short estrous cycle and that superovulation can be induced. However, biological characterization of the gametes in this species has not been done. The purpose of this study is to determine requirements for fertilization and to devise a suitable method for in vitro fertilization (IVF). Data from these studies show that 1) cauda epididymal plasma (CEP) is required for maintaining sperm motility in vitro; 2) in a heterologous sperm penetration assay in vitro, optimal conditions for sperm capacitation and the acrosome reaction depend on time and CEP concentration; and 3) loss of the cortical granules is observed to occur spontaneously within 3-4 h after ovulation in superovulated eggs. On the basis of the data obtained from these studies, we have developed a method for preincubation of mastomys sperm and a successful mastomys IVF procedure. This species can now be used as a new laboratory animal for research in reproductive biology.

Acrosome↗

Neuroepithelial and ependymal changes in HTX rats with congenital hydrocephalus: an ultrastructural and immunohistochemical study.

To investigate the pathogenesis of congenital hydrocephalus the brains of HTX rats aged between 16 days and 4 weeks and the brains of normal Wistar rats of the same ages were examined. In the fetal HTX rat brains, the lateral ventricles were symmetrically dilated from 20 days of gestation. The neuroepithelium bordering the ventricles showed thinning with cellular disarrangement and deformity. Similar neuroepithelial abnormalities were also found in the lateral ventricles of the HTX rat brain with no macroscopic signs of hydrocephalus at 20 days of gestation. The neuroepithelium showed flattening of the cells, widening of the intercellular spaces, formation of microvilli on the detached lateral cell surfaces, and frequent macrophage infiltration. On the other hand, the neuroepithelial cells of the third ventricle and the aqueduct were affected less severely or showed no significant abnormalities. Immunohistochemically, most of the neuroepithelium and ependyma of the lateral ventricles were positive for vimentin in both prenatal and postnatal hydrocephalic HTX rats, while a small number or none of those in normal control rats were positive. These morphological changes suggested that preferential involvement of the lateral ventricular neuroepithelium might be closely associated with the pathogenesis of congenital hydrocephalus in HTX rats.

Aging↗

So-called neoplastic myoepithelial cells in chondroid syringomas/mixed tumors of the skin: their subtypes and immunohistochemical analysis.

An immunohistochemical study of nine cases of chondroid syringomas/mixed tumors of the skin was performed to elucidate the nature of the so-called neoplastic myoepithelial cells (NMEC) in tumor tissues. These nine tumors contained NMEC of considerable variability in number from one tumor to another. These NMEC were classified into three types: (i) hyaline cells (plasmacytoid cells); (ii) spindle NMEC; and (iii) polyhedral cells. They showed different immunostaining patterns, as the following describes. Cytokeratin 14 was positive in most of the spindle NMEC and a large number of the polyhedral cells, and in a small number of the hyaline cells. Concerning low molecular weight cytokeratins, most of the hyaline cells showed immunoreactivity, whereas they were negative in many of the spindle NMEC and were expressed only in a small number of the polyhedral cells. alpha-Smooth muscle actin and muscle-specific actin were positive in the spindle NMEC but negative in any of the hyaline cells and polyhedral cells. These findings strongly indicate that the hyaline cells and the spindle NMEC are of the simple epithelial and myoepithelial types, respectively. The findings also suggest that the polyhedral cells show differentiation toward basal cells of the sweat gland dermal ducts or myoepithelial cells.

Adenoma, Pleomorphic↗

Sexual difference in early fetal crown-rump length versus gestational age in pregnancies arising from in vitro fertilization.

The purpose of this study was to investigate the influence of fetal sex on the crown-rump length (CRL) derived from pregnancies resulting from in vitro fertilization without the uncertain conception dating in the first trimester. With ultrasonography, we performed 334 and 353 examinations, on 88 male and 102 female fetuses, respectively. The relationship between gestational age and fetal CRL was explored with regression analysis: male CRL (mm) = 1.13 x gestational age (days) - 47.39 (r = 0.97), female CRL = 1.11 x gestational age - 46.44 (r = 0.97). These regression lines were mutually included in the 95% confidence intervals for each other. Our results indicate no statistically significant difference in the CRL between the two sexes, being supportive of using the identical fetal CRL criterion regardless of fetal sex for gestational age assessment with ultrasonography in the first trimester of pregnancy.

Crown-Rump Length↗

Development and sexual difference in embryonic heart rates in pregnancies resulting from in vitro fertilization.

The purpose of this study was to investigate the gestational change of embryonic heart rates (EHRs) and to estimate the influence of embryonic sex on the EHR in pregnancies resulting from in vitro fertilization in the early first trimester. With transvaginal ultrasonography, we performed 92 and 105 examinations, on 27 male and 30 female embryos, respectively. The EHR increased gradually from 87 beats per min at 38 days of gestation to 189 beats per min at 62 days of gestation. The relationship between gestational age and EHR was explored by regression analysis: male EHR (beats per min) = 3.78 x gestational age (days) - 51.30 (r = 0.95), female EHR = 3.65 x gestational age - 44.56 (r = 0.95). These regression lines were mutually included in the 95% confidence intervals for each other. Our results indicate both a close positive correlation between the EHR and gestational age and no statistically significant difference in the EHR between male and female embryos. These findings suggest that the EHR measurement is a novel method for very early ultrasound dating with the identical EHR criterion without regard to embryonic sex.

Embryonic and Fetal Development↗

Development of hepatic sinusoidal structure with special reference to the Ito cells.

To elucidate sinusoidal cell structure and function under normal conditions and their behavior in diseased settings, an understanding of their developmental aspects is needed. At day 10 of gestation in mice and rats or at 5 weeks of gestation in humans, the hepatic cords grow into the mesenchymal tissue of the septum transversum, and the primitive sinusoidlike structure is simultaneously observed between the liver cell cords. In the margin of the growing liver primordium, mesenchymal cells in the septum transversum are trapped in the subendothelial space. These subendothelial cells are at the early stages of organogenesis and become progenitors of the Ito cells. By days 12-14 of gestation in mice and rats or 8 weeks of gestation in humans, the basic structure of the sinusoids has developed. Embryonic hepatic sinusoids are usually lined by a continuous endothelium without basement membranes, and an incompletely fenestrated sinusoid appears at the middle gestational stage. In the late gestational stages, the Ito cells exhibit myofibroblastlike features in humans, mice, and rats. In association with this event, perisinusoidal reticular networks are gradually intensified. After birth until days 4-5 in mice and rats, the sinusoidal and perisinusoidal structures are almost completely formed, although slight morphological differences from those in adult livers still exist. What happens to sinusoidal endothelial cells and Ito cells in hepatic fibrosis-cirrhosis of the adult may be a deviated or uncontrolled occurrence of what goes on during the fetal period, i.e., a continuous nonfenestrated sinusoidal lining in the early embryonic stage and a myofibroblastlike transformation of Ito cells in late fetal life.

Animals↗

Aging-related changes in calcium oscillations in fertilized mouse oocytes.

Aging of oocytes, being not fertilized after ovulation for a prolonged time, considerably affects normal development of the fertilized oocyte. We examined effects of the aging on a series of highly repetitive Ca2+ transients commonly seen in fertilized mouse oocytes (Ca2+ oscillations). Frequency of Ca2+ oscillations in the aged oocyte [20 hrs after induction of superovulation by i.p. human chorionic gonadotropin (hCG)] was significantly higher (34.1 +/- 5.8 l/hr) than the fresh oocyte (14 hr post-hCG, 21.8 +/- 7.9 l/hr). Rates of rise and fall of individual Ca2+ transient in the aged oocyte were significantly slower than the fresh oocyte, whereas durations of individual Ca2+ transients were similar. When extracellular Ca2+ was raised from 2.04 mM to 5.00 mM, aged oocytes showed significant prolongation of the duration of individual Ca2+ transient, that resulted in a sustained elevation of intracellular Ca2+ ([Ca2+]i) in 33% of the aged oocyte. Transient increase in [Ca2+]i by photolysis of a caged Ca2+, Nitr-5, injected into cytoplasm was completely restored in the fresh oocyte [fluorescence intensity of [Ca2+]i indicator dye Fluo-3 (F480) returned to 97 +/- 2% of the control level, time constant = 37 +/- 9 sec]. In contrast, in the aged oocyte, restoration of F480 following Nitr-5 photolysis was incomplete (115 +/- 12% of the control) and slow (time constant = 64 +/- 23 sec). Because inhibition of the Ca2+ pump of the endoplasmic reticulum (ER) by 5 microM thapsigargin almost completely inhibited restoration of F480 following Nitr-5 photolysis in the fresh oocyte, we conclude that the aging-related changes in Ca2+ oscillations may be accounted for by dysfunction of intracellular Ca2+ regulation, presumably of the Ca2+ pump of the ER.

Aging↗

The incidence of apoptotic bodies in membrana granulosa can predict prognosis of ova from patients participating in in vitro fertilization programs.

OBJECTIVE: To investigate the relationship between the incidence of apoptotic bodies in membrana granulosa and follicular steroid concentrations in human follicles. DESIGN: Case-controlled prospective study for 132 individual follicles. SETTING: Procedures were performed in Yamagata University Hospital. PATIENT(S): Thirty-six normo-ovulatory women with tubal infertility underwent ovulation induction for IVF-ET with a conventional hyperstimulation method. INTERVENTION(S): Patients underwent follicle aspiration after the administration of hCG. MAIN OUTCOME MEASURE(S): The nuclei of recovered granulosa cells were examined by fluorescence microscopy, and the incidence of apoptotic bodies was tabulated. Intrafollicular steroids were evaluated mainly by RIA. These data were analyzed with respect to oocyte-retrieval, oocyte maturity, fertilization, and embryo quality. RESULT(S): Membrana granulosa cells in the follicles from which oocytes were subsequently fertilized showed a significantly lower incidence of apoptotic bodies than those in follicles from which the oocytes did not fertilize. Membrana granulosa cells in the follicles from which oocytes were developed into good quality showed a significantly lower incidence of apoptotic bodies than those in the follicles from which oocytes developed into fair and poor quality. The incidence of apoptotic bodies was significantly higher in the mural granulosa cell region than in the cumulus cell region in most cases. Intrafollicular E2, P, and free T levels were not different between the oocyte groups. CONCLUSION(S): These results indicate that lower incidence of apoptotic bodies in individual follicles is associated with better outcomes for oocytes. Also, mural granulosa cells and cumulus cell in each follicle may show differentiation during follicular maturation.

Apoptosis↗

Incidence of apoptotic bodies in membrana granulosa of the patients participating in an in vitro fertilization program.

OBJECTIVE: To investigate the incidence of apoptotic bodies in mural granulosa cell masses and cumulus cell masses. DESIGN: Nonrandomized, prospective study. SETTING: Department of Obstetrics and Gynecology, Yamagata University School of Medicine, Yamagata, Japan. PATIENT(S): One hundred twenty-nine normally ovulating women underwent ovulation induction for IVF-ET with GnRH analogue (GnRH-a) and gonadotropins. INTERVENTION(S): Patients underwent follicle aspiration after the administration of hCG. MAIN OUTCOME MEASURE(S): The nuclei of recovered granulosa cells were examined by fluorescence microscopy and the incidence of apoptotic bodies was tabulated. RESULT(S): The incidence of apoptotic bodies was significantly higher in mural granulosa cell masses than in cumulus cell masses in the entire group of 129 patients. Both incidence of apoptotic bodies of mural granulosa cell masses and cumulus cell masses were significantly higher in patients with less than six follicular oocytes compared with patients with six or more oocytes. Nonpregnant patients showed significantly higher incidence of apoptotic bodies in mural granulosa cell masses compared with pregnant patients. CONCLUSION(S): These results indicate that mural granulosa cell masses and cumulus cell masses may have different functions in follicular maturation. The incidence of apoptotic bodies in mural granulosa cell masses can be used as an indicator of success of IVF.

Adult↗

The optimal equilibration time for mouse embryos frozen by vitrification with trehalose.

To determine the best equilibration time in the cryoprotectant before rapid cooling, 8-cell mouse embryos were exposed to a vitrification solution containing ethylene glycol, Ficoll and trehalose in modified phosphate-buffered saline at 5 degrees C for varying periods of time. They were frozen using an ultra rapid freezing method, thawed in a 20 degrees C water bath and cultured for 24 h with 5-bromo-2-deoxyuridine. Embryo development and the number of sister chromatid exchanges, a sensitive indicator of genetic damage, were observed. The results demonstrated that embryo development after freezing and thawing was similar among the groups exposed for periods of 5-40 min. However, the number of sister chromatid exchanges was significantly smaller in the group exposed for 5 min, indicating that this was the safest equilibration time in the vitrification solution.

Animals↗

Stress of female infertility: relations to length of treatment.

The psychology of infertile women was investigated with a battery of psychological tests consisting of a semistructured interview, State Trait Anxiety Inventory (STAI), Center for Epidemiologic Studies Depression Scale (CES-D), and Cornell Medical Index (CMI). The subjects were 107 infertile women being treated for infertility. The semistructured interviews revealed that the stress factor for infertile women changes with the length of infertility. In the early states, the main stress is related to a physical inferiority complex, while later it changes into stress about what others outside the family say. According to STAI, CES-D and CMI, infertile women are considered to become more depressive the longer treatment persists. Therefore, counseling for infertile women should be adapted to long-term treatment.

Adult↗