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Biomedical subjects

T Motoyama

Publications and source records attributed to T Motoyama.

At least 181 records · Page 10Linked to original sources

[The effect of enzymes on progesterone-receptor binding and chromatin binding of the complex in the estrogen-primed rabbit uterus (author's transl)].

The present study was designed to determine the characteristics of the progesterone receptor and chromatin binding site ("acceptor") of the progesterone-receptor complex in the rabbit uterus. The uterus was obtained from an estrogen-primed immature female rabbit. The binding of progesterone to the uterine receptor was examined in vitro. The progesterone-receptor binding was reduced only by proteases, and phosphorus moiety may not be related for progesterone-receptor binding. The effects of enzymes on the acceptor of the chromatin were investigated. The progesterone-receptor complex was bound to the dehistonized chromatin. The dehistonized chromatins, which were pretreated with enzymes at 4 degrees C or 37 degrees C for 30 minutes, were incubated with 3H-progesterone prelabeled uterine cytosol at 4 degrees C for 30 minutes, and the radioactivity in the chromatin pellet was counted. Proteases effectively decreased the receptor binding capacity to the dehistonized chromatin in the following order: pronase greater than trypsin greater than papain greater alpha-chymotrypsin. DNAse moderately and phospholipase A slightly decreased its binding capacity. The results may indicate that the acceptor site of the progesterone receptor is nonhistone protein over DNA of chromatin and may contain phosphorus moiety.

Animals↗

Preliminary studies on streoid-binding proteins in human testes of testicular feminization syndrome.

This study was designed to detect either 5alpha-dihydrotestosterone (DHT) or 17beta-estradiol (E2)-binding protein in the testes of a 1-year-old patient with testicular feminization syndrome (TFS) and in the testes of patients with prostatic cancer. Sucrose gradient analyses revealed E27S protein binding (but no such 7S protein binding of DHT) in the testes of the patient with TFS, but both E2 and DHT 7 S protein binding was observed in normal senile testes. The dissociation constants (Kd) were measured by charcoal adsorption. The Kd of E2 protein binding in both testes of different status was approximately 1.3 x 10(-9) M, and the Kd of DHT protein binding was 2.0 x 10(-9) M in the senile testes. A ligand specificty study indicated characteristics of both E2 and DHT receptors in the senile testes. It is speculated that a deficiency of androgen receptor and the presence of estrogen receptor in the testes of patients with TFS lead to insensitivity to androgen as a result of the androgen receptor deficiency and to sensitivity to estrogen as a result of the presence of the estrogen receptor.

Androgen-Insensitivity Syndrome↗

[Interaction of norethindrone on estrogen and progesterone receptors in the rabbit uterine cytosol (author's transl)].

Norethindrone (ENT), which is a representative in estrane series of progestogen, is not only strongly progestational but also estrogenic and in some cases, antiestrogenic. To understand progestational effect and antiestrogenic effect, the interactions of ENT on estrogen and progestogen receptors were studied in the uterine cytosol of white female rabbit. The 274,200 X G supernatant of uterine homogenate was used as cytosol. 3H-Estradiol, 3H-Progesterone, 3H-ENT or cold ENT were incubated with uterine cytosol at 4 degrees C for 2 hours. Results are as follows: 1. Sucrose gradient centrifugation [5 approximately 20% linear and 40,000 rpm (159,200 X G) for 16 hours at 4 degrees C]: ENT was bound to extrogen 8S receptor in immature rabbit uterus (Fig. 2 & 3), and to progestogen 8S receptor in estrogen primed rabbit uterus (Fig. 5). 2. Kinetic study, determined by dextran coated charcoal (0.001% dextran and 0.1% charcoal): (1) In the uterine cytosol of immature rabbit, 3H-estradiol-receptor binding was observed with Kd divide by 3.6 X 10-9 M and it was revealed that ENT was a competitive inhibitor to this binding with Ki divide by 2.6 X 10-6 M, as in Fig. 6. (2) 8S component, obtained by centrifugation of uterine cytosol (Fig. 1) in estrogen primed rabbit, binds 3H-progesterone with Kd divide by 8.1 X 10-10 M and Bm (maximal binding sites) divide by 5.0 X 10-8 M/mg of protein, and ENT was a competitive inhibitor in this binding with Ki divide by 2.3 X 10-9 M (FIG. 7 & 8). 3H-ENT-8S binding was demonstrated with Kd divide by 1.1 X 10-9 M and Bm divide by 8.7 X 10-8 M/mg of cytosol protein (Fig. 8). These results indicate: (a) ENT is bound to both estrogen and progestogen receptors in 8S macromolecules of uterine cytosol, (b) competitive inhibition of ENT to these bindings indicated that ENT is bound to these receptors at the steroid binding sites where estradiol and progesterone bind to, (c) ENT has much more affinity to progestogen receptor (Ki divide by 2.3 X 10-9 M) than to estrogen receptor (Ki divide 2.6 X 10-6 M), (d) while ENT is bound to progestogen and estrogen receptors at the same time, Bm of ENT (8.7 X 10-8 M/mg of cytosol protein) is more than Bm of progesterone (5.0 X 10-9 M/mg of cytosol protein), and Kd of ENT (1.1 X 10-9 M) was less than Ki of ENT (2.3 X 10-9 M) in the binding to progesterone-receptor. Biologically, while ENT is bound to progestogen -receptor with high affinity and to estrogen receptor with low affinity, ENT is actually progestational in low dose and antiestrogenic in high dose but the anti-estrogenicity seems to be incomplete in vivo as ENT may be metabolized to a potent estrogenic compound, ethinyl estradiol

Animals↗

Separation and detection of bile acid 3-glucuronides in human urine by liquid chromatography/electrospray ionization-mass spectrometry.

A method for the separation and detection of bile acid 3-glucuronides by liquid chromatography (LC)/electrospray ionization (ESI)-mass spectrometry (MS) has been developed. On the ESI mode, glucuronides were characterized by an intense pseudo-molecular ion [M-h]- with a doubly charged ion [M-2H]2-, and the ratio of these negative ions were markedly influenced by an acidic component of salt added to a mobile phase, according to a pKa value of an acidic moiety at C-24. Bile acid 3-glucuronides in human urine were extracted with a SepPak C18 cartridge, followed by purification on lipophylic ion exchange gel, piperidinohydroxypropyl Sephadex LH-20. Subsequent resolution into unconjugated, glycine and taurine conjugated bile acid 3-glucuronides was attained by LC on a Develosil ODS-HG-5 column using 20 mM ammonium acetate (pH 7.0)-methanol (10:12, v/v) as a mobile phase, where 3-glucuronides were monitored with a characteristic negative ion [M-H]-. The newly developed method was applied to the quantitatiation of bile acid 3-glucuronides in human urine with satisfactory sensitivity and reliability.

Bile Acids and Salts↗

Mixed gonadal dysgenesis: endogenous hormonal effects in the endometrium and histogenesis of germinoma.

The clinicopathologic features of two patients with mixed gonadal dysgenesis are presented, with specific reference to the relationship between endogenous sex hormones and the endometrium and the development of neoplastic disease. One patient, whose immature gonad contained granulosa cells and theca cells, had elevated serum estrogen levels and an endometrium with frequent ciliated metaplasia and squamous metaplasia. Another patient had elevated serum testosterone levels and atrophic endometrium. Both had gonadal tumors, more specifically, germinomas, which contained many calcified nodules within the tumor. These findings suggest that these germinomas arose from a gonadoblastoma.

Adult↗

Extremely well differentiated squamous cell carcinoma of the breast. Report of a case with a comparative study of an epidermal cyst.

BACKGROUND: Although the cytologic diagnosis of malignancy by fine needle aspiration is easy in cases of moderately to poorly differentiated squamous cell carcinoma in various organs, it is not always easy in cases of the well-differentiated type. CASE: A 46-year-old female presented with a slowly growing mass in the right breast; it was diagnosed as an extremely well differentiated squamous cell carcinoma. The diagnosis of malignancy was initially not established by fine needle aspiration cytology because of minimal cellular atypia. The smear contained many squamous cells, mainly of the superficial type, with little cellular atypia, in a background of horny material. However, neither anuclear superficial cells nor granular cells were found. CONCLUSION: Absence of anuclear keratinized cells and granular cells is probably a helpful finding for distinguishing squamous cell carcinoma from epidermal cyst in the breast region.

Biopsy, Needle↗

Ascitic fluid cytologic features of a malignant mixed mesodermal tumor of the ovary.

A case of malignant mixed mesodermal tumor of the ovary in a postmenopausal patient presenting with abdominal distension is reported. Cytologic examination of smears of the ascitic fluid showed the presence of adenocarcinomatous and sarcomatous cells (with some of the latter being giant cells) plus numerous unidentifiable cells that bore some resemblance to either mesothelial cells or macrophages. Electron microscopic studies showed a clear differentiation of the adenocarcinomatous and sarcomatous cells from positively identified mesothelial cells and macrophages also present in the ascitic specimen, indicating that the unidentified cells in fact originated in the adenocarcinoma (endometrioid carcinoma), chondrosarcoma and unclassified sarcoma found in the surgical specimen. The differential diagnostic cytomorphologic and electron microscopic features are described in detail.

Adenocarcinoma↗

Immunohistochemical properties of malignant mesothelioma cells in histologic and cytologic specimens.

Although there is general agreement that immunohistochemical methods can aid in the pathologic diagnosis of malignant mesothelioma, some studies have produced conflicting results. To obtain comparable and reproducible results, unequivocal malignant mesotheliomas were studied with the biotin-streptavidin-peroxidase complex method in 14 formalin-fixed, paraffin-embedded tissue specimens, 5 ethanol-fixed smear slides and 3 cold acetone-fixed smear slides. The expression of CA-125, epithelial membrane antigen (EMA) and vimentin by malignant epithelial mesothelioma cells was hindered by their poor preservation in formalin fixative. Cytologic specimens fixed in cold acetone were the best type for immunohistochemistry. The majority of malignant epithelial mesothelioma cells in the smear slides fixed in cold acetone were positive for CA-125, EMA, low-molecular-weight cytokeratin and vimentin, but none of them were positive for carcinoembryonic antigen, CA-19-9, epithelial antigen, high-molecular-weight cytokeratin or Leu-M1.

Biomarkers↗

Fine needle aspiration cytology of primary mediastinal germ cell tumors.

Fine needle aspiration cytology was performed on six patients with malignant mediastinal germ cell tumor: 1 pure seminoma, 1 pure embryonal carcinoma, 1 pure yolk sac tumor and 3 mixed germ cell tumors containing teratoma. Their cytologic features were compared with each other and with the cytologic features of thymoma, which arises commonly in the anterior mediastinum. A definitive cytologic diagnosis could be made only in the cases of seminoma because of its characteristic cytologic features. Cytochemical staining for germ cell alkaline phosphatase was helpful in diagnosing seminoma in the cytologic examination, while the presence of hyaline globule or alpha-fetoprotein immunostaining as the cytologic diagnostic feature of yolk sac tumor were not necessarily found in fine needle aspiration cytology. All epithelial, lymphocytic and mixed type thymomas were easily differentiated from the four types of germ cell tumor examined.

Adolescent↗

Mucinous cystic tumor of the retroperitoneum. A report of two cases.

Two cases of retroperitoneal mucinous cystic tumors and some diagnostic problems on aspiration cytology are presented. The first was a mixed benign, borderline and malignant mucinous tumor, and the second was a mixed benign and borderline mucinous tumor. Our knowledge of the cytology of ovarian mucinous cystic tumors was not helpful in making a correct diagnosis in these cases. Based on our experience, we emphasize the following. First, consider methods of collecting enough cells to make a diagnosis. Second, consider the possibility of mixed histologic features. Direct aspiration, if possible, from papillary lesions inside the cyst will probably lead to an accurate diagnosis. Measurement of carcinoembryonic antigen levels in the cystic fluid is useful in checking for underdiagnosis of such tumors.

Adult↗