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T Fukaya

Publications and source records attributed to T Fukaya.

At least 91 records · Page 5Linked to original sources

Substance P induces inositol 1,4,5-trisphosphate and intracellular free calcium increase in cultured normal human epidermal keratinocytes.

Substance P is a neuropeptide which is present in peripheral C nerve endings and released from them. Free nerve endings of C nerve are present in human epidermis. The effects of substance P on the transmembrane signaling system of pig epidermal sheets were previously reported. In these studies, a small amount of cells other than keratinocytes contaminated the epidermal sheets and the species difference from human was also noticed. Therefore we investigated the effects of substance P on cultured normal human epidermal keratinocytes. Alteration of intracellular free calcium (Ca2+) in single living keratinocytes was studied using an inverted fluorescence microscope and Ca(2+)-sensitive dye, Fura 2-AM. Treatment of normal human epidermal keratinocytes with substance P resulted in an increase in inositol 1,4,5-trisphosphate and in intracellular Ca2+. Substance P inhibited DNA synthesis of the keratinocytes in a dose-dependent manner. These results are consistent with the view that substance P stimulates phosphatidylinositol-4,5-bisphosphate hydrolysis of human keratinocytes, resulting in inositol 1,4,5-trisphosphate-Ca2+ signal.

Calcium↗

Quantitation of P450 aromatase immunoreactivity in human ovary during the menstrual cycle: relationship between the enzyme activity and immunointensity.

To understand changes associated with the menstrual cycle in the human ovary, it is very important to examine chronological changes in P450 aromatase (P450arom) enzymatic activity in the normal cycling ovary. Therefore, we initially examined the correlation between intensity of P450arom immunoreactivity and its biochemical enzymatic activity in five estrogen-producing human cancer cell lines (HHUA, Ishikawa, HEC-59, OMC-2, and MCF-7). P450arom immunointensity per cell was evaluated by the CAS 200 computed image analysis system, and its catalytic activity per 10(6) culture cells was analyzed by the tritiated water method. A significant correlation (r = 0.959) was demonstrated between P450arom immunoreactivity and enzymatic activity under optimal conditions of tissue fixation and immunohistochemical procedures. We then investigated P450arom immunointensity in 31 specimens of normal cycling human ovaries to examine chronological changes in P450arom activity per cell throughout the menstrual cycle. In the follicular phase, P450arom was observed in the granulosa cells of one selected antral follicle per case during the mid- to late proliferative period, and its immunointensity per granulosa cell in the follicle was not significantly different between mid- and late proliferative periods, although serum estradiol level was markedly elevated in the late proliferative period. In the luteal phase, both P450arom immunointensity per luteinized granulosa cell in a corpus luteum and serum estradiol level reached a peak in the mid-secretory period. These findings indicate that different factors may influence ovarian P450arom activity during the follicular and luteal phases, i.e., an increased number of granulosa cells in the selected follicle during the follicular phase but changes in P450arom activity per luteinized granulosa cell in the corpus luteum during the luteal phase.

3,3'-Diaminobenzidine↗

Influence of HLA antigens on reproduction among Japanese population: study of haplotypes in 247 families.

Although an increased incidence of shared HLA antigens among couples experiencing recurrent spontaneous abortion or infertility has been observed, antigenic effects remain to be clarified. These effects, which are suspected to exist among fertile couples, were examined in this study. A group of 247 healthy Japanese families and their 682 children, whose HLA antigen haplotypes could be determined, were studied. Haplotypes of the fathers were compared with those of the mothers, and the combination of HLA antigens in the A, B, C and DR loci was determined as antigenic patterns against the mother. Patterns inherited by children were examined and deviations of the patterns were studied according to the natural rule that either of the haplotypes should be inherited at equal rates. Lower rates of children with antigens common with those of the mother, which may show a result of reproductive selection, were observed among only the first born children in the A (p < 0.05) and DR loci (p < 0.01), but rates similar to those expected were observed among the second born and later children. Lower rates were also observed in the B and C loci among all children with no significant difference between groups of the children. The necessity of differing antigenic factors in order to achieve pregnancy in the population was shown. Important effects of HLA antigenicity in reproductive failure are suggested.

Adult↗

Is treatment of long-term and consecutive use of clomiphene citrate effective in anovulatory patients? Results of multi-centric retrospective studies.

The efficacy of long term consecutive treatment of clomiphene citrate (CC) was investigated in 608 infertile women associated with anovulatory disorders (classified as WHO group II amenorrhea). This study was performed in multi-centric facilities of obstetrics and gynecology by a retrospective randomized assessment. Infertile women associated with anovulation were treated by 50-150 mg of CC for five consecutive days in each treatment cycle. In this study, 200 pregnancies were observed and the total pregnancy rate was 32.9%. The number of abortions were 33 out of the 200 pregnancies (16.5%). The cumulative pregnancy rate within the pregnant subjects reached 90% in initial 10 treatment cycles and there was no difference between 50 mg and 100 mg of CC treatment. There were remarkable differences among the pregnancy rates achieved during consecutive treatments at the primary (clinics and hospital), secondary (regional center hospitals) and advanced facilities. Consecutive treatment by CC for infertile women associated with anovulation may be effective during initial 10 cycles. Although ovulation can be confirmed during the following cycles, the stimulation protocol should be reconsidered after 12 consecutive cycles therapy with CC, because cycle fecundity decreases. Hence, to improve pregnancy rates, it is important to check the previous administration of CC which reflects adversely on the subsequent conception rate especially in advanced facilities where these patients are referred to.

Anovulation↗

The role of macrophage colony stimulating factor in the peritoneal fluid in infertile patients with endometriosis.

Macrophage colony stimulating factor (M-CSF) concentrations in the peritoneal fluid of 44 infertile patients were measured. The concentration of M-CSF was significantly higher in the patients with endometriosis than in those without endometriosis. There was no significant difference between the M-CSF concentration and stage of endometriosis classified by revised American Fertility Society (rAFS) classification in the 44 patients. However, in primary infertility patients with endometriosis there was a significant correlation between peritoneal M-CSF levels and rAFS points of endometriosis. We speculate that there is some relationship between the increased levels of M-CSF in the peritoneal cavity and the primary infertility associated with endometriosis especially progression of endometriosis.

Ascitic Fluid↗

Laparoscopic evaluation of the onset and progression of endometriosis.

OBJECTIVE: To clarify the pathogenesis of endometriosis on the basis of analysis of primary lesion sites, age at onset, rate of progression, and response to drug treatment. STUDY DESIGN: The clinical records of 690 women with laparoscopically confirmed endometriosis were retrospectively analyzed based on the revised American Fertility Society point system. RESULTS: The primary site of endometriosis was the uterosacral ligament and pelvic peritoneum/pouch of Douglas in 73% of patients with stage I disease, whereas only 16% had ovarian lesions. However, disease progression was associated with an increasing frequency of ovarian lesions. In terms of the revised American Fertility Society score, endometriosis progressed at a mean rate of 0.3 point per month. Thus the earliest onset of endometriosis was estimated at 3 to 4 years after menarche. Drug therapy improved the revised American Fertility Society score by about 50%. Patients with a low response to an initial cycle of therapy generally showed further improvement after an additional treatment cycle. CONCLUSIONS: Because endometriosis may occur as early as 3 to 4 years after menarche and gradually progresses, drug therapy, including long-term treatment, should be carried out in women with definitive evidence of endometriosis who must maintain their reproductive potential.

Adolescent↗

Is pelvic endometriosis always associated with chronic pain? A retrospective study of 618 cases diagnosed by laparoscopy.

OBJECTIVE: To establish a relationship between pelvic pain and the stage and severity of pelvic endometriosis determined according to the revised American Fertility Society classification. STUDY DESIGN: Retrospective study of 618 cases diagnosed by laparoscopy as endometriosis at the Tohoku University Hospital from 1984 to 1991. RESULTS: Pelvic pain did not appropriately reflect the stage and severity of pelvic endometriosis in 40% of patients. Although the pain in some patients in stage IV showed some correlation with the stage and severity of the disease, others had either no symptoms or paradoxically complained of less pain than those patients with stage I or II endometriosis. CONCLUSIONS: With the present revised American Fertility Society classification of endometriosis we cannot evaluate the severity of pelvic pain.

Chronic Disease↗

Temporal and spatial localization of steroidogenic enzymes in premenopausal human ovaries: in situ hybridization and immunohistochemical study.

In situ hybridization and immunohistochemical localization of cytochrome P450 cholesterol side-chain cleavage (P450scc), 3 beta-hydroxysteroid dehydrogenase (3 beta HSD), cytochrome P450 17 alpha-hydroxylase (P450c17) and cytochrome P450 aromatase (P450arom) was performed in 50 morphologically normal human premenopausal ovaries, and correlated these findings with their endometrial phase. In general, mRNA expression of these enzymes examined by in situ hybridization were in good agreement with immunolocalization examined by immunohistochemistry. Expression of P450scc, 3 beta HSD and P450c17 was observed in large-sized preantral follicles, consisting of more than five layers of granulosa cells, preovulatory follicles, corpora lutea, and some degenerating corpora lutea and atretic follicles in all endometrial phases. Several follicles and/or corpora lutea positive for these enzymes were observed in the same ovary. Expression of P450arom was generally observed in only one follicle (antral or preovulatory follicle) or corpus luteum per case in mid proliferative to premenstrual phase, and was not observed in menstrual to early proliferative phase. These findings indicated that (1) expression of steroidogenic enzymes was associated with the continual human ovarian process including follicular development and atresia, and (2) especially, P450arom expression may occur only in a selected antral follicle and may have an important role in dominant follicular development.

3-Hydroxysteroid Dehydrogenases↗

Distribution of tenascin in normal cycling human ovary.

Tenascin is an extracellular matrix glycoprotein, which has been reported to be involved in parenchymal-mesenchymal interactions during morphogenesis, wound healing, and carcinogenesis. Tenascin immunolocalization was performed in 51 specimens of morphologically normal human ovaries by using a specific monoclonal antibody against purified human fibroblast tenascin. In preovulatory follicles, no significant immunoreactivity was detected. In functioning corpora lutea, immunoreactivity was present as a fine border around the periphery. In association with the involution of the corpora lutea, marked diffuse tenascin immunostaining in the intercellular space was observed. These data raise the question of whether tenascin may be involved in luteolysis and may play an important role in the ovarian cycle by regulating the involution of corpora lutea.

Cell Adhesion Molecules, Neuronal↗

Compression of the left brachiocephalic vein: cause of high signal intensity of the left sigmoid sinus and internal jugular vein on MR images.

To study the asymmetry in signal intensity of the sigmoid sinuses, internal jugular veins (IJVs), or both sets of structures on magnetic resonance (MR) images, the authors reviewed 226 serial sets of routine MR imaging studies. Cerebral digital subtraction angiography was performed in 20 patients with a markedly higher intensity and/or enhancement on the left; 15 of them also underwent venography. With every sequence, the left sigmoid sinus, IJV, or both were higher in signal intensity significantly more often than the right (range, P = .0001 to .0129). Angiography revealed hemostasis in the left IJV in 16 patients that disappeared during full inspiration in 14. In 13, venography revealed that the left brachiocephalic vein was compressed to occlusion between the aortic arch and the sternum during tidal volume ventilation. The occlusion disappeared at full inspiration, when the distance between the aortic arch and the sternum increased. This hemostasis could be the major cause of the frequent increased signal intensity of the left sigmoid sinus and IJV on MR images.

Adolescent↗

The role of inhibin on pituitary FSH secretion and maturation of pituitary-gonadal axis in immature female rats.

The possible physiological significance of endogenous inhibin was evaluated in prepubertal female rats during sexual maturation. Ovarian/serum inhibin, serum FSH levels were measured from 4th day to 35th day of life by RIA. Serum inhibin levels were first detected on 4th day of life, thereafter, progressively increasing up to 27th day. Serum FSH levels gradually increased up to the 14th day with a sharp fall in the FSH levels on day 15 which afterwards was maintained at a lower level. Pregnant mare serum gonadotropin (PMSG) treatment on day 14 and day 22 rats showed different patterns of serum FSH and inhibin changes. In 14 day group, there was no difference between control and PMSG treatment, but in 22 day group, not only significant difference between control and PMSG treatment but also inverted relations between FSH and inhibin after PMSG treatment were found. It can be postulated from these findings that secretion of inhibin from the granulosa cells is initiated by FSH by its direct action on the ovary whereas the level of FSH itself is controlled by inhibin after FSH reaches peak values. Also it can be inferred that the feedback control mechanism of inhibin seems to be established around 3rd week of life.

Animals↗

[Cochlear nerve conduction block restoration of hearing after removal of cerebellopontine tumors].

Hearing restoration after removal of cerebellopontine tumors has only infrequently been observed. We present 5 cases with dramatic improvement of both pure tone threshold and speech threshold after excision of tumors. Two patients had acoustic neuroma, 2 meningioma and one cerebellar hemangioblastoma. Preoperatively, the patients suffered severe sensorineural hearing loss and had no measurable speech discrimination score. Recordings of the auditory brain stem response were absent or showed only wave I. However, results of the electrocochleogram were within normal limits. After removal of the tumors, their hearing returned to normal and remained stable thereafter. The data are consistent with the hypothesis that pressure from the tumor causes cochlear nerve conduction block and hearing impairment.

Adult↗

Triple cancers in the urogenital area of a patient with aplastic anemia.

Three epithelial neoplastic lesions, perineal Bowenoid papulosis, uterine cervical carcinoma, and bladder transitional cell carcinoma, which occurred in a mildly immunosuppressed patient who had aplastic anemia were studied for human papillomavirus (HPV) infection. In the Bowenoid papulosis, HPV type 16 DNA was identified by polymerase chain reaction (PCR) and by in situ hybridization (ISH). In contrast, in the uterine cervical carcinoma, HPV 16 was not detected, although possibly another unidentified type of HPV in the lesion was suggested by the ISH findings. In the bladder transitional cell carcinoma, neither papillomavirus genus-specific (PGS) antigen nor HPV DNA was found.

Adult↗

Adenylate cyclase induces intracellular calcium increase in single human epidermal keratinocytes measured by fluorescence microscopy using Fura 2-AM.

Intracellular calcium ([Ca2+]i) is an important second messenger of extracellular signals to induce various cellular responses. Extracellular and intracellular Ca2+ are considered to be important for cellular differentiation and proliferation of epidermal keratinocytes. Several mechanisms which increase [Ca2+]i have been demonstrated in various tissues, but in epidermal keratinocytes these mechanisms are poorly understood. In epidermal keratinocytes the adenylate cyclase-cyclic AMP response is thought to regulate cell proliferation and differentiation. However, the series of reactions which follow the cyclic AMP response remain unknown. Beta-adrenergic agonists increase [Ca2+]i in cultured epidermal keratinocytes, and we have therefore studied whether stimulation of keratinocyte adenylate cyclase could induce [Ca2+]i increase, by using fluorescence microscopy with Fura 2-AM. Adenosine and histamine, which are known to be keratinocyte adenylate cyclase receptor agonists, induced transient [Ca2+]i increase, as did epinephrine. In addition, forskolin, a direct adenylate cyclase activator, and dibutyryl-cyclic AMP also induced an increase in [Ca2+]i. In a calcium-free medium epinephrine, adenosine, histamine and dibutyryl-cyclic AMP induced an increase in [Ca2+]i. These results suggest that cyclic AMP in human epidermal keratinocytes regulates [Ca2+]i, which is released from intracellular stores.

Adenosine↗

Substance P induces intracellular calcium increase and translocation of protein kinase C in epidermis.

Substance P is a neuropeptide present in, and released from, peripheral C nerve endings. The presence of substance P-positive nerve fibres in the epidermis has been reported. We investigated the effect of substance P on the transmembrane signalling system of pig epidermal keratinocytes. Treatment of pig epidermis with substance P resulted in an increase in inositol 1,4,5-trisphosphate (IP3), and in intracellular free calcium. The treatment also resulted in translocation of protein kinase C from a cytosol to a membrane fraction. Substance P, however, did not affect the beta-adrenergic- or histamine (H2)- adenylate cyclase responses of the epidermis. Neither forskolin-induced, nor cholera toxin-induced cyclic AMP accumulation were affected by substance P treatment. These results consistent with the view that substance P stimulates phosphatidylinositol-4,5-bisphosphate (PIP2) hydrolysis of keratinocytes, resulting in IP3-Ca2+ and diacylglycerol-protein kinase C signal activation. Although protein kinase C is known to affect the epidermal adenylate cyclase system, no evidence for such 'cross-talk regulation' was detected in keratinocytes by substance P treatment.

Animals↗

The spontaneous LH surge in ovarian hyperstimulation for assisted reproductive technology.

Thirty-seven patients who underwent assisted reproductive technology programme, in vitro fertilisation or embryo transfer and gamete intra-fallopian transfer, were randomised into two groups in order to evaluate frequency of premature spontaneous LH surge when stimulated with different type of hMG. An hMG (FSH:LH = 1:1) was administered for eighteen patients (group A) and the different highly purified hMG (FSH:LH = 19:1) was administered for nineteen patients (group B). Blood samples are drawn from day 3 of the cycle until hCG administration, and serum LH level are measured in the frozen samples. There were no statistical difference in the total amount of hMG used before hCG injection, days of hMG injection and the number of oocytes retrieved between two groups. The premature spontaneous LH surge occurred only one patient in group A (5.6%), but more frequent LH surge was observed in group B (42.1%) (p < 0.05). It is suggested that the different FSH/LH ratio may be the reason for the difference of the incidence of premature spontaneous LH surge.

Chorionic Gonadotropin↗

Adenylate cyclase induces intracellular Ca2+ increase in single human epidermal keratinocytes of the epidermal sheet as measured by digital imaging microscopy using Fura 2-AM.

Intracellular Ca2+ ([Ca2+]i) is thought to act as a second messenger of transmembrane signalling systems. However, no measurement of [Ca2+]i has been made in intact epidermal keratinocytes. We have developed a method for measuring [Ca2+]i in human keratinocytes from pure epidermal sheet by the application of digital imaging fluorescence microscopy with the use of Fura 2-AM. Normal human pure epidermal sheets were obtained by dispase treatment. Epinephrine and salbutamol induced transient [Ca2+]i increases. Propranolol, a beta-antagonist, inhibited this response, while prazosin and yohimbine (alpha 1- and alpha 2-antagonists, respectively) did not affect the response. Histamine and adenosine, also receptor agonists of the epidermal adenylate cyclase system, induced a similar [Ca2+]i increase, as did forskolin, a direct activator of adenylate cyclase. These data coincide with those previously presented for cultured human epidermal keratinocytes, and reveal that adenylate cyclase activation induces an increase of [Ca2+]i in intact epidermal cells. This technique enables the kinetics of [Ca2+]i in various skin disorders to be investigated.

Adenosine↗