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Biomedical subjects

M Nohara

Publications and source records attributed to M Nohara.

At least 37 records · Page 2Linked to original sources

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↗

Altered calcium handling in compensated hypertrophied rat cardiomyocytes induced by pressure overload.

To investigate Ca2+ handling in compensated hypertrophied cardiomyocytes, we measured Ca2+ transients and contraction of hypertrophied rat left ventricular myocytes induced by aortic constriction (AC). The fluorescence ratio (I405/I480) after indo-1/AM loading and circumferential length were simultaneously measured in isolated myocytes. The amplitude of Ca2+ transients (Ca-Amp) was higher in rats with AC than in sham-operated rats (Sham) (0.25 +/- 0.08 vs 0.17 +/- 0.05). There was a positive correlation between Ca-Amp and fractional shortening (FS) in both AC and Sham rats, whereas the ratio of FS/Ca-Amp was smaller in AC rats. These observations suggest that compensated hypertrophied cardiomyocytes exhibit an adaptive increase in Ca-Amp, associated with reduced myofilament responsiveness to an increase in Ca2+. Isoproterenol and forskolin increased Ca-Amp and FS, and decreased time to 50% decline of Ca2+ transients. Although myocytes from AC rats exhibited reduced responsiveness to isoproterenol, responses to forskolin did not differ between the 2 groups. The reduced beta-adrenergic response in Ca2+ handling was probably due to altered beta-adrenoceptor numbers, G-protein function and/or their coupling process.

Adrenergic beta-Agonists↗

Electron microscopic immunohistochemistry of ocular and extraocular pseudoexfoliative material.

To investigate the nature of extraocular pseudoexfoliative (PSX) material, and to validate the hypothesis that the PSX syndrome is a systemic disorder, we studied the immunoreactivity of intraocular and extraocular PSX material from patients with the PSX syndrome using antibodies against proteins of extracellular matrices. Surgical specimens of four different tissues were obtained from human eyes with the PSX syndrome: nine trabecular tissues, three cataractous lenses, six bulbar conjunctivas, and seven lid skins. These tissues were processed for electron microscopic immunohistochemistry, and stained with antibodies against vitronectin, fibronectin, laminin, and elastin, by an indirect immunogold procedure. Density of the gold particles located on the PSX material was estimated by video image analysis. Statistical analysis of the data revealed that the PSX material from trabecular tissues, lenses, bulbar conjunctivas, and lid skins had almost identical immunoreactivity to the antibodies studied. PSX materials of trabecular tissues, lenses, bulbar conjunctivas, and lid skins were found to have not only a similar ultrastructure, but also almost identical immunohistochemical characteristics. These results support the hypothesis that the PSX syndrome is a systemic disorder.

Aged↗

Preschool boys and girls use no differently.

Twenty-one videotape recorded interactions of 42 preschool children (aged 3;4 to 5;3) in same-sexed dyads were coded and analysed to see how these children used the word no in their interactions. Results showed that while boys and girls used the word no equally frequently, they were found to use the word in different ways. A trend for boys to use no when correcting or prohibiting their playmate's behaviour more often than girls was revealed. Girls were more likely to provide reasons for denying or rejecting their playmate's proposition or suggestion. The overall findings of this study are comparable to previously reported results indicating that males and females employ different styles of speech in order to accomplish the same goal. However, unlike most studies conducted on language and sex, this study offers a unique approach in that it examines how boys and girls use a word that occurs equally frequently in their vocabulary to accomplish different goals.

Child, Preschool↗

Sarin poisoning in Matsumoto, Japan.

A presumed terrorist attack with sarin occurred in a residential area of the city of Matsumoto, Japan, on June 27, 1994. About 600 residents and rescue staff were poisoned; 58 were admitted to hospitals, and 7 died. We examined clinical and laboratory findings of 264 people who sought treatment and the results of health examinations on 155 residents done 3 weeks after the poisoning. Findings for severely poisoned people were decreases in serum cholinesterase, acetylcholinesterase in erythrocytes, serum triglyceride, serum potassium and chloride; and increases in serum creatine kinase, leucocytes, and ketones in urine. Slight fever and epileptiform abnormalities on electroencephalogram were present for up to 30 days. Examination revealed no persisting abnormal physical findings in any individual. Acetylcholinesterase returned to normal within 3 months in all people examined. Although subclinical miosis and neuropathy were present 30 days after exposure, almost all symptoms of sarin exposure disappeared rapidly and left no sequelae in most people.

Acute Disease↗

Electron microscopic lectin histochemistry of the trabecular meshworks in human eyes.

Ten normal human trabecular meshworks were examined by electron microscopy using avidin-biotin complex in order to investigate the localization of binding sites of eight lectins. The tissue specimens were fixed in paraformaldehyde and glutaraldehyde mixture and embedded in Lowicryl K4M at low temperature. The ultrathin sections were stained with biotin labelled lectins and colloidal gold labelled streptoavidin and were observed with the conventional transmission electron microscope. Some lectins such as ABA, ConA and DSA were localized on fine fibrils underneath the endothelium of the trabecular wall of the Schlemm's canal, electron-dense cores of elastic fibers, fine granular like materials, basement membranes, collagen fibers and the long-spacing fibers. However, the other lectins such as DBA, SBA, Lotus, UEA-I and RCA60 were not specifically localized in these tissues. From the results it was demonstrated that the differential ultrastructural localization of glycoconjugate residues in the human trabecular meshworks can be revealed using this lectin staining.

Adult↗

Lectin electron microscopic histochemistry of the pseudoexfoliative material in the skin.

PURPOSE: To investigate the hypothesis that pseudoexfoliation (PSX) syndrome is a systemic disorder, the authors studied the composition of glycoconjugates in the intraocular and extraocular PSX material at the electron microscopic level, using a panel of lectins as cytochemical probes. METHODS: The authors examined 8 lid skins, 11 trabecular tissues, and 3 cataractous lenses from human eyes with PSX syndrome. Tissues were processed for electron microscopical histochemistry and stained with PNA, RCA120, DBA, SBA, ConA, WGA, UEA-I, and Lotus, with an indirect lectin-colloidal gold technique. RESULTS: Both the intraocular and extraocular PSX materials manifested almost identical reactivity to lectins, which indicated that glycoconjugates in the PSX material contained with sugar residues of galactose (PNA, RCA120), alpha-mannose (ConA), and N-acetyl-D-glucosamine (WGA). On the other hand, it was indicated that sugar residues of N-acetyl-D-galactosamine (DBA, SBA) and fucose (UEA-I, Lotus) were absent. Granular inclusions and microfibrils in the capsule and ocular zonules were stained similarly and weakly. CONCLUSIONS: The intraocular and extraocular PSX materials contained the same sugar residues of glycoconjugates, which suggested that those materials had the same nature. This study, the first documentation of lectin-binding sites on the extraocular PSX material, supported the hypothesis of PSX syndrome as a systemic disorder.

Exfoliation Syndrome↗

Cumulus mass maintains embryo quality.

OBJECTIVE: To determine if co-culture of early stage embryos with their own cumulus mass improves embryo quality. DESIGN: Before insemination, cumulus masses along with cumulus matrices were separated from the oocytes surrounded by a few remaining cumulus cells. Fourteen hours after insemination, fertilized oocytes were each placed onto the cumulus cells, and matrix and co-culture commenced. The embryos were observed every 24 hours. Fifty-three oocytes were treated in co-culture (C) and 59 oocytes were treated in routine culture (U). RESULTS: Thirty-four (C) and 43 (U) oocytes were fertilized and placed on growth media for further culture. Twenty-four hours after culture, 10 embryos (29%) in C and 12 (28%) in U were good quality, and 4 embryos (12%) in C and 7 (16%) in U were of poor quality. Seventy-two hours after culture, 10 (29%) in C and 8 (18%) in U were of good quality, and 3 (9%) in C and 13 (30%) were of poor quality. The percentage of good quality embryos in the co-culture group was significantly higher than in the control group after 72 hours. Conversely, the percentage of poor quality embryos in the co-culture group was significantly lower than that in the control group after 72 hours. CONCLUSION: Co-culture maintains embryo quality over prolonged culture times. This facilitates the development of good quality embryos for ET.

Cells, Cultured↗

Localization of elastin in the normal and glaucomatous human trabecular meshwork.

PURPOSE: The extracellular materials (ECMs) in the trabecular meshwork (TM) are thought to play a crucial role in aqueous outflow resistance. Immunohistochemical localization of elastin, one of the major ECMs in the normal and glaucomatous human TM, was examined ultrastructurally. METHODS: Eight normal eye bank eyes and 16 trabeculectomy specimens of primary open angle glaucoma (POAG, 11 eyes from 8 cases), congenital glaucoma (2 eyes from 1 case), and juvenile glaucoma (3 eyes from 2 cases) were embedded in Lowicryl K4M at low temperature. The distribution of elastin was studied by the protein A-gold technique. RESULTS: In normals, the gold particles indicating the antigenic sites for elastin existed mainly in the central amorphous element of the elastic-like fibers, and a few gold particles were observed within the area containing fine granular-like material and fine fibrillar-like material. No labeling was observed in cellular materials or other ECMs. In congenital and juvenile glaucoma, labeling was similar to that observed in normals. In POAG specimens compared to normals, there was an increased amount of elastin-bound immunogold particles along the inner canal endothelium. The increased gold particles, which did not have a fibrillar arrangement and were not enclosed by electron-dense microfibrils, were found within the area containing fine fibrillar-like material. However, labeling within the elastic-like fibers was similar to that observed in normals. CONCLUSIONS: Under electron microscopy, elastin could be localized in the normal and glaucomatous human TM. The results of this investigation suggest that elastin may play an important role in the etiology of POAG.

Adult↗

Dominance of activated T cells and interleukin-6 in aqueous humor in Vogt-Koyanagi-Harada disease.

PURPOSE: To determine the immunopathologic role of the lymphocytes and lymphokines in aqueous humor (AH) of patients with Vogt-Koyanagi-Harada disease (VKH). METHODS: The distribution of leukocyte subsets in the peripheral blood and AH was examined using fluorescein isothiocyanate-conjugated monoclonal antibodies. The levels of lymphokines, such as interleukin-2 (IL-2) and interleukin-6 (IL-6), in the sera, AH, and cerebrospinal fluid from the patients with VKH were determined using an enzyme-linked immunosorbent assay. RESULTS: T cells constituted the majority of lymphocytes within AH. The value for CD4+ cells (helper/inducer T lymphocytes) in AH was 51.7% +/- 14.9% (mean +/- SD) and that for CD8+ cells (cytotoxic/suppressor T lymphocytes) was 31.1% +/- 13.0%. The percentage of HLA-DR+ cells (B lymphocytes, monocytes, macrophages, and activated T lymphocytes) in AH (50.8% +/- 24.9%) significantly exceeded (P < 0.001) that in blood (13.1% +/- 4.2%). The percentage of CD8+ cells in AH from three patients with the delayed type of VKH rose during their clinical course. The level of IL-6 was significantly elevated in AH from the patients with VKH. The level of IL-6 in AH correlated with the number of lymphocytes in AH, and it reflected the severity of the inflammatory response in AH of patients with VKH. The level of IL-2 in the sera, AH, and cerebrospinal fluid was in the normal range. CONCLUSIONS: Aqueous humor lymphocytes from the patients with VKH were more activated than were peripheral blood lymphocytes. IL-6 may play an important role as an inflammatory mediator in VKH. It may be useful to analyze the lymphocyte subsets and the levels of lymphokines, especially of IL-6, at the site of inflammation in uvea to improve the criteria for assessing the prognosis of VKH.

Adult↗

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↗

[Lectin histochemical studies on species differences in the mammalian trabecular meshwork].

Lectin histochemical studies were performed to clarify the species differences in the location of glycoconjugates in the trabecular meshwork (TM) of various mammals, i.e., the mouse, rat, rabbit, pig, and ox, to find an experimental model for human TM. Cryosections were made and stained with sixteen kinds of biotinylated lectin followed by an avidin-biotin-peroxidase complex (ABC). Strong positive reactions in the TM of all 5 kinds of mammals were observed for ConA, PHA-E4, PHA-L4, WGA, ABA, LCA, RCA 60, RCA 120, DSA, and SSA. The TM of the 5 kinds of mammals was weakly positive for Lotus. Rabbit, pig and ox TM were positive for MAM and others were negative. Rabbit, pig and ox TM were weakly positive for PNA and were negative in the other's. Rat TM was weakly positive for SBA and was negative in the other's. The TM of all 5 kinds of mammals was negative for UEA-I and DBA. It could be concluded that species difference exists in lectin binding site in the TM.

Animals↗

Decision making in psychiatric civil commitment: an experimental analysis.

OBJECTIVE: Legislation in Canada and the United States that was intended to decrease the use of civil commitment has resulted in a paradoxical increase in involuntary hospital admissions. To elucidate the reasons for this increase, this study was designed to assess the relative importance of various factors involved in the decision to commit a patient. METHOD: All psychiatrists in Ontario were sent a questionnaire asking them to make commitment decisions based on hypothetical case vignettes. Four factors were systematically varied in the vignettes: the patients' legal commitability, clinical treatability, alternative resources, and psychotic symptoms. Completed questionnaires, with three vignettes each, were returned by 495 respondents. RESULTS: All four variables were statistically significant in the expected direction; legal commitability (i.e., dangerousness to self and/or others, inability to care for self) and presence of psychotic symptoms accounted for the majority of the variance in the final decision to commit. CONCLUSIONS: These results suggest that psychiatrists in Ontario rely primarily on legally mandated factors (i.e., psychosis and dangerousness) in making their decisions to commit, although a considerable amount of individual variation is also evident.

Adult↗