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Biomedical subjects

H Suginami

Publications and source records attributed to H Suginami.

At least 55 records · Page 3Linked to original sources

A factor inhibiting ovum capture by the oviductal fimbriae present in endometriosis peritoneal fluid.

The cumulus-fimbria interaction was investigated in vitro with endometriosis peritoneal fluid (PF). On the oviductal fimbria of the golden hamster, incubated with endometriosis and nonendometriosis PF and culture medium 199 contained 4% bovine serum albumin (control), was placed the mouse oocyte-cumulus complex at 5-minute intervals for 60 to 70 minutes. Incubation periods until the loss of fimbrial capability of ovum capture (ovum capturability disappearance time [OCDT] ) were 23.3 +/- 2.7 (standard error of the mean), 51.7 +/- 2.9, and 61.3 +/- 0.9 minutes with endometriosis, nonendometriosis PF, and control, respectively. OCDT was significantly decreased with cell-free and ultrafiltrated endometriosis PF containing a molecular size greater than 100,000 (26.0 +/- 2.8 and 26.1 +/- 2.5 minutes, respectively). A factor inhibiting fimbrial capability of ovum capture is present in endometriosis PF.

Adult↗

Ovulation induction with bromocriptine in normoprolactinemic anovulatory women.

Afternoon-evening and nocturnal serum PRL levels and PRL responsiveness to metoclopramide (MCP) were determined in 34 women with normoprolactinemic anovulation (nPRL-Anov) and in the early follicular phase (EFP) in 6 normal women. Subsequently, the nPRL-Anov women were treated with 5 mg bromocriptine (Br) twice daily for 2 months, and its action on ovulation was determined. Those women who did not respond to Br received 50-150 mg clomiphene for 5 days. The nPRL-Anov patients were classified into 3 groups in terms of the efficacy of Br treatment: group I, those who ovulated with Br (n = 13); group II, those who ovulated after receiving Br and clomiphene (n = 7); and group III, those who failed to ovulate after the above treatments (n = 10). Four patients dropped out of the study. Diurnal serum PRL levels were approximately 10 ng/ml in all patients, and no statistical difference was found among the groups. Peak nocturnal serum PRL levels (the highest PRL value during the 0000-0400 h period) were 38.0 +/- 23.9 (+/- SD) ng/ml in group I patients, higher (P less than 0.05) than in groups II and III and normal (EFP) women (20.1 +/- 9.1, 20.7 +/- 7.7, and 16.3 +/- 2.7 ng/ml for the group II and III patients and the EFP women, respectively). MCP induced rapid and marked elevation in serum PRL levels in all subjects. The maximum post-MCP PRL value in the group I patients was 224.2 +/- 89.7 ng/ml, which was significantly higher (P less than 0.002) than the maximum value in the remaining groups (120.5 +/- 25.8, 121.3 +/- 54.2, and 101.9 +/- 28.1 ng/ml, respectively). Ten (76.9%) and 12 (92.3%) group I patients had nocturnal PRL levels above 25 ng/ml and maximum PRL values after MCP above 150 ng/ml, respectively. We conclude that some nPRL-Anov patients have elevated nocturnal serum PRL levels or enhanced PRL responsiveness to MCP, indicative of nocturnal or latent hyperprolactinemia. Br effectively induced ovulation in these patients. A MCP provocation test can predict the outcome of Br treatment in such nPRL-Anov patients.

Adult↗

[Serum prolactin levels during sleep and in metoclopramide stimulation in normoprolactinemic anovulation and ovulation induction with bromocriptine].

Diurnal and nocturnal serum prolactin (PRL) levels were determined in 37 normoprolactinemic anovulatory (nPRL-Anov) patients and 6 normal cycling women each in the early follicular phase (EFP) and the luteal phase (LP). PRL responsiveness to metoclopramide (MCP) was assessed. The nPRL-Anov patients received 5mg/day bromocriptine (CB-154) for 2 months and ovulation was investigated. To the CB-154 non-responders was added 150mg/day clomiphene (CL) for 5 days. The nPRL-Anov patients were classified into 3 groups; i.e. I: those who ovulated with CB-154 (n = 14), II: those who ovulated with CB-154 + CL (n = 6), and III: those who failed to ovulate (n = 10). Nocturnal serum PRL levels in I (42.0 +/- 26.2ng/ml; M +/- SD) were significantly higher than those in EFP, II and III (p less than 0.05). Eleven (78.6%) of I had nocturnal PRL levels higher than 25ng/ml. The peak PRL values at MCP provocation were significantly higher in I (213.3 +/- 89.1ng/ml) than in EFP, II and III (p less than 0.01). Thirteen (92.9%) of I had peak PRL values more than 150ng/ml. The peak PRL values and nocturnal PRL levels correlated. There were some exhibiting nocturnal or latent hyperprolactinemia, who responded effectively to CB-154, among nPRL-Anov patients. A MCP provocation test could be used in prediction of the efficacy of CB-154 treatment when applied to nPRL-Anov patients.

Anovulation↗

Qualitative and quantitative differences in hLH species in the first and second LH release induced by continuous stimulation with synthetic LHRH in normal menstrual cycle as assessed by isoelectrofocusing.

It is known that continuous stimulation with LHRH induces a biphasic release of LH. The two pools theory is generally accepted in interpreting this phenomenon. The present study was conducted in order to elicit qualitative differences in LH included in the two pools. Five hundred micrograms synthetic LHRH was infused for 2h in 10 women in various phases of the menstrual cycle and 5 h blood sampling was performed during and after the infusion. Biphasic release of LH was demonstrated in all the cases investigated. The responsiveness to LHRH was most predominant in the midcycle, followed by the luteal phase. Pools of plasma samples at 30-105 min and 150-225 min of the LHRH infusion (plasma pools I and II; corresponding to the initial and the second LH release, respectively) were subjected to isoelectrofocusing (IEF) fractionation, pH range of 3.5-10. LH in plasma was classified in terms of isoelectric point (pI); i.e. A(pI = 9.13), B(8.60), C(8.16), D(7.67), E(7.24), and F(LH species migrating in the acidic pH area). Percentage of high alkaline LH species was significantly elevated in plasma pool I of the midcycle. No phasic difference was observed in the IEF profiles in plasma pool II. Consistent and significant increase and decrease in the percentage of acidic and high alkaline LH species, respectively, were observed throughout the menstrual cycle when the IEF profiles of plasma pool II were compared with those of pool I. The ratio of biological to immunological LH activities (B/I ratio) was markedly depressed in acidic LH species. The highest B/I ratios were obtained in the LH species migrating in the mid-alkaline region. The acidic LH species might represent the young generation of LH molecules prior to acquisition of biological potency. LH species migrating in the mid-alkaline region might be mature and bio-potent forms of the hormone. High alkaline LH species might represent LH molecules over-processed and having lost a degree of biological potency during a prolonged period of storage.

Adult↗

A case of endometriosis of the lung treated with danazol.

A 25-year-old Japanese woman, complaining of catamenial hemoptysis and severe cough complicated with menorrhalgia, was diagnosed as having pulmonary and pelvic endometriosis. She was treated with danazol for 20 weeks. Significant improvement of her condition was achieved during the treatment period. Catamenial hemoptysis recurred at the first menstruation after termination of the treatment. Readministration of danazol was refused. Therefore, surgical removal of the affected lobe of the right lung was performed. Cases of this rare disorder are reviewed.

Adult↗

Subpopulations of luteinizing hormone (LH) possessing various ratios of bioactivity to immunoreactivity in the female rat pituitary glands and their changes during the estrous cycle.

Twenty-four adult female Sprague-Dawley rats (3 from each of 8 litters), showing 4-day cycles, were used in the present study. Aqueous extracts of pools of 6 pituitary glands in each cycle date were fractionated with a column isoelectrofocusing (IEF) technique, pH range of 3.5-10. Biological and immunological LH activities were determined by an in vitro bioassay and a radioimmunoassay, respectively, in the original aqueous extracts of the pituitary glands and in the fractions separated by IEF. Pituitary content of LH was the highest in the proestrus before the preovulatory LH surge (1243.7 +/- 67.8 micrograms NIAMDD rat LH-RP-1/pituitary gland for the biological activity). In the estrus, after the LH surge, it was reduced to 688.9 +/- 51.2 micrograms/pituitary gland. The decreased pituitary content was recovered to the level in the proestrus during the metestrus and the diestrus (1047.0 +/- 53.8 and 1173.0 +/- 58.5 micrograms/pituitary gland, respectively). Rat LH in the pituitary aqueous extracts was separated into multiple subpopulations in terms of pI values by IEF; i.e. Subpopulations A (pI = 10.3), B (9.3), C (9.0), D (8.7), E (8.3), F (neutral LH), and G (acidic LH). Among them the most predominant one was Subpopulation A throughout the estrous cycle. Subpopulations A, B and C exhibited statistically significant cyclic changes as was observed in the pituitary LH content, whereas the remaining ones stayed at constant levels during the cycle. The highest ratio of biological to immunological LH activities (B/I ratio) was obtained in Subpopulation A (6.41), followed by G, C and B (5.15, 4.24 and 3.99, respectively). Depressed B/I ratios were revealed in D, E and F (2.59, 1.86 and 3.07, respectively). High alkaline LH subpopulations, i.e. A, B and C, preserving high biological potency and showing cyclic changes during the estrous cycle, seem to be the releasable types of the hormone and to be mainly discharged for the preovulatory LH surge. Although characteristic features of other types of the hormone are not known, it is possible that one of them, presumably the acidic LH, might be the newly-synthesized type of the hormone, which might attain releasability by certain molecular modifications involving a shift in the pI value.

Animals↗

Qualitative and quantitative changes in plasma luteinizing hormone (LH) under stimulation by intravenous infusion of synthetic luteinizing hormone-releasing hormone (LHRH) in Japanese monkeys (Macaca fuscata) as assessed by electrofocusing.

Using 5 adult Japanese monkeys (Macaca fuscata) of both sexes, the qualitative and quantitative changes in plasma LH caused by LHRH administration were examined before and after castration. Five hundred micrograms synthetic LHRH was infused for 2 hrs and blood samples were obtained every 15 min for 5 hrs under ketamine hydrochloride anesthesia. Plasma LH levels were determined by an in vitro bioassay using the mouse testis interstitial cell preparation. The LHRH infusion induced a biphasic release of LH in intact animals of both sexes and in castrated females but not in castrated males. The biphasic pattern of LH release was characterized by an initial acute rise followed by a plateau or a fall and the consecutive second rise after a long latent period. Aliquots of plasma samples obtained during 30-90 (corresponding to the initial rise) and 120-210 min (corresponding to the second rise) after the initiation of LHRH infusion were pooled separately (plasma pools I and II, respectively) and subjected to isoelectrofocusing (IEF) fractionation. The IEF fractionation was performed in ampholine-sucrose gradient, pH range 3.5-10, under a constant electrical power supply (3 W, 700 V) for 48 hrs at 4 degrees C. After the completion of the IEF fractionation the LH activity in each fraction was measured by the in vitro bioassay. The IEF fractionation separated plasma LH into 6 distinct species in terms of pI values. When the IEF profiles of plasma pools I and II in intact animals of both sexes were compared, the increase and the decrease in acidic to neutral LH and alkaline LH, respectively, were consistent and significant in plasma pool II. Changes of the same kind were further exaggerated when the IEF profiles in castrated animals were compared with those in non-castrated ones. The acidic LH may represent the LH species newly synthesized and not well processed. The LH species possessing pI values of 7.5-8.5 is likely to be the mature type of LH exhibiting high biological potency. The LH species migrating in the high alkaline region seems to be the LH of over-maturation.

Animals↗

Immunohistochemical localization of a human chorionic gonadotropin-like substance in the human pituitary gland.

A substance with the biological, immunological, and physicochemical properties similar to those of hCG (hCG-like substance) has been found in human pituitary homogenates and urine of nonpregnant subjects. The purpose of the present study was to localize the hCG-like substance in human pituitary gonadotropin producing cells (gonadotrophs) by using an unique antiserum raised against a synthetic hCG beta C-terminal peptide (P75R). Twenty-five pituitary glands were obtained at autopsy from women of various chronological ages. An immunohistochemical method was employed for localizing hLH, hFSH, and an hCG-like substance on the horizontal serial sections of the pituitary glands. The LH and FSH cells were indistinguishable with the antisera employed in the present study. The hCG-like substance was only localized in the cytoplasm of the gonadotrophs, rather commonly when the pituitary glands from postmenopausal women were examined. The P75R serum pretreated with excessive amount of native hCG failed to exhibit the immunostaining. Normal human tissues aside from pituitary glands did not react with the P75R serum. The results obtained in the present study and those reported by others may indicate that the hCG-like substance present in human pituitary glands is the precursor of hLH.

Adolescent↗

Biological and immunological characterization of human luteinizing hormone discharged in a pulsatile fashion in the normal menstrual cycle.

Biological and immunological measurements of human luteinizing hormone (hLH) were performed with plasma samples obtained by continuous withdrawal over a 10-hour period in various phases of the normal menstrual cycle. A pulsatile pattern of hLH discharge was demonstrated by both the in vitro bioassay and radioimmunoassay throughout the menstrual cycle, although the frequency and the amplitude of the pulse differed according to the menstrual stage. The decreased half disappearance time of immunoreactive hLH (83.7 min with 95% confidence limits at 64.3 and 103.1 min), in comparison with that of bioactive hLH (122.9 (98.2: 147.6) min), and the rapid recovery of the depressed biological to immunological activity ratio (B/I ratio) at the peak of the pulse (0.87 (0.77:0.96) in the periovulatory phase indicated the concomitant discharge of hLH subunits with hLH. The differences in the half disappearance times of bioactive hLH in the follicular and luteal phases (103.6 (94.7:112.5) min and 87.2 (72.5: 101.9) min, respectively) and in the B/I ratios at the peaks of the pulses in these two phases (1.17 (1.05:1.30) and 1.03 (0.94:1.12), respectively) suggested the selective secretion of hLH subpopulations with various biological and immunological potencies and with various clearance rates. The significantly depressed B/I ratios at the trough in the luteal phase (0.58 (0.44:0.76) were sufficient to question the overall reliability of the hLH-RIA procedure employed, especially when applied to low hLH levels, even after several methodological improvements.

Adult↗

Biological and immunological characterization of human luteinizing hormone discharged by the stimulation of synthetic luteinizing hormone-releasing hormone (LH-RH) in normal and anovulatory women.

Plasma samples were obtained by repeated venopunctures immediately before, and at 15, 30, 60, 120 and 180 min after intravenous bolus administration of 100 micrograms synthetic LH-RH in normal and various anovulatory women. Plasma hLH levels were determined by an in vitro bioassay and a radioimmunoassay with improved reagents. The LH-RH stimulation induced an abrupt elevation of both biological and immunological hLH activities in normal and anovulatory women, although the responsiveness to LH-RH differed from case to case. Both elevated biological and immunological hLH activities decreased gradually with a half disappearance time of 122.4 +/- 27.9 min and 112.5 +/- 25.4 min, respectively (Mean +/- S.D.). A transient and significant depression in the ratio of biological to immunological hLH activities (B/I ratio) was observed at 15 min after the LH-RH administration in normal subjects. This depression is attributable to the cross-contamination of an increased amount of hLH subunits induced by LH-RH stimulation. The B/I ratios were significantly elevated throughout the investigation period in the anovulatory patients when compared with those in the normal subjects. This elevation appears to indicate the increased discharge of special type(s) of hLH subpopulations of high biological potency in the anovulatory cases.

Adult↗

Measurements of human luteinizing hormone in plasma by in vitro bioassay and by conventional and improved radioimmunoassays.

Daily plasma samples obtained from normally menstruating women were measured for human luteinizing hormone (hLH) by in vitro bioassay and by conventional and improved radioimmunoassays (RIA). The improvement of hLH RIA consisted in the choice of standard and the extensive purification of tracer. The profiles of the biological and immunological activity of hLH throughout the cycle showed a qualitative similarity with the mid-cycle hLH surge. The estimated values obtained with the conventional RIA were consistently and significantly higher than those by the improved RIA (p less than 0.001). The elevation of the Y-intercept on RIA (5.22 +/- 3.22 uIU/ml) indicated the possible influences of crossreacting substances on the estimates obtained with the conventional RIA. The Y-intercept values calculated from the bioassay and the improved RIA estimates were indistinguishable from the origin (0.45 +/- 1.81 mIU/ml). The inter-individual variations of the biological/immunological activity ratios indicate the different proportions of multiple sub-populations of hLH molecules with various biological and immunological activities in each case. The improved RIA was concluded to be more preferable for detecting hLH levels in biological fluid than the conventional RIA. The need for more suitable standard preparations for hLH RIA was proposed.

Adult↗

A trial for the improvement of the in vitro bioassay for human luteinizing hormones.

In order to improve the sensitivity and the reliability of the in vitro bioassay for human luteinizing hormone, various strains of male mice at various chronological ages were sacrificed to obtained dispersed testes interstitial cell preparations. They consisted of DDN, DDY, DBA, C57, C3H, ICR, and BAL/C at 5, 7 and 9 weeks of age. The C57 Black strain, especially at 7 and 9 weeks of age, gave the most satisfactory standard dilution curve with a sensitivity of less than 12.5 microIU/tube, with a wide assayable range and with a parallelism to the sample dilution curve. Improvement of the assay is expected with the use of the C57 Black strain as the source of the cell suspension.

Animals↗

Influence of the purity of the iodinated tracer on the specificity of the radioimmunoassay of human follicle-stimulating hormone.

The objective of this study was to establish whether [125I]hFSH subunits are formed during iodination of hFsh by a mild (lactoperoxidase) technique, and if so which fractionation procedure provides the best separation of [125I]hFSH from its subunits. In addition, the effect of the 125I subunits on the specificity of the hFSH radioimmunoassay technique was examined. The products of iodination of hFSH were fractionated by a number of techniques consisting of low and high resolution gel filtration procedures and adsorption on either cellulose of Concanavalin A coupled to Sepharose. The efficacy of the adsorption methods was assessed by a subsequent high resolution gel filtration procedure (Ultrogel AcA 54). Radioreceptor and radioimmunoassay methods were used to monitor the profiles of hFSH, hFSH alpha and hFSH beta subunits and to assess the recoveries of radioreceptor-active hFSH obtained by the different procedures. Iodination of a highly purified hFSH preparation invariably resulted in the formation of both [125I]hFSH subunits. The proportion of radioactivity associated with the 125I subunit peak to that of the [125I]hFSH peak was 10% when the iodination products after low resolution gel filtration were fractionated by a high resolution gel filtration procedure. The corresponding amount of subunits obtained by adsorption on cellulose and Concanavalin A was 71% and 6%, respectively. Thus in comparison with low resolution gel filtration there was a partial separation of [125I]hFSH from [125I]hFSH subunits on Concanavalin A while no separation was effected on cellulose. In fact, the results suggest that 125I subunits were formed during fractionation on cellulose. The recoveries of hFSH activity following fractionation on cellulose and Concanavalin A were significantly lower (27% and 65%) than those obtained by either low or high resolution gel filtration techniques. A marked reduction ;n the specificity of the RIA of hFSH was found when [125I]hFSH subunits rather than [125I]hFSH were used as tracer in the assay. It is concluded that the presence of 125I subunits in [125I]hFSH preparations used as tracer in the RIA of hFSH can diminish the specificity of the assay. The routine employment of a high resolution gel filtration procedure which provides a complete separation of 125I subunits from the intact hormone, is suitable for the elimination of this source of assay invalidity while the other fractionation procedures tested (cellulose adsorption or Concanavalin A) are not.

Adsorption↗

Studies on a human chorionic gonadotrophin-like material present in non-pregnant subjects.

The presence of an hCG-like material in urinary and pituitary extracts and plasma obtained from non-pregnant subjects was investigated. Two assay methods were used to detect this material following fractionation of pituitary and urinary extracts by gel filtration (Ultrogel AcA 54) and/or isoelectrofocusing: a) a radioimmunoassay employing an antiserum raised against a specific sequence of the carboxy terminal region (residues 115--145) of the beta-subunit of hCG, and b) an in vitro bioassay method which measures both hLH and hCG activities. The fractionation procedures employed provide a satisfactory separation of highly purified hCG and hLH preparations. In the pituitary and urinary extracts hCG beta-peptide-like immunoactive (PIA) material was found consistently, which co-eluted with iodinated hCG following gel filtration and possessed pI values similar to those of hCG when subjected to isoelectrofocusing. The PIA material also exhibited in vitro biological activity similar to that shown by hLH and hCG. Detectable levels of immunoactive material were also found in plasma; however, the plasma levels of this PIA material were not influenced by classical endocrine measures such as the stimulation or inhibition of gonadotrophin secretion. The low levels of this material in plasma precluded its further characterization by gel filtration or electrofocusing. Whereas the present data and those reported by other investigators seem to suggest the presence of some hCG-like material in urinary and pituitary extracts and possibly in plasma of non-pregnant subjects, it is emphasized that the available evidence is not sufficiently conclusive to exclude other interpretations as to the nature of this material.

Biological Assay↗

Influence of the purity of the iodinated tracer on the specificity of the radioimmunoassay of human luteinizing hormone.

Human luteinizing hormone (HLH) iodinated with 125I by the use of a lactoperoxidase method, was fractionated by either cellulose adsorption, gel filtration or by the combination of these methods. The products of iodination were characterized by their in vitro biological LH activity and by their binding profiles with antisera to HLH, HLHalpha and HLHbeta subunits. Several radioactive components were obtained after gel filtration with or without an initial cellulose adsorption step. One of these fractions was identified as biologically active HLH and another as the HLHalpha subunit. Radioimmunoassay studies were conducted with different iodinated fractions as tracers, using three well defined and widely available antisera to HLH. The standard used was the HLH International Reference Preparation for immunoassay (68/40). Cross-reactivity was examined with several purified pituitary preparations, such as HFSH, HTSH, HLHalpha and HLHbeta subunit. A significantly higher cross-reactivity with HLHalpha, HFSH and HTSH was obtained with the [125I]HLHalpha fraction as tracer than with biologically active [125I]HLH. Furthermore, in the radioimmunoassay of HLH preparations of varying purity, significantly higher estimates of immunological activity were obtained with the [125I]HLHalpha tracer than with the biologically active [125I]HLH. It is concluded that the presence of [125I]HLHalpha in the [125I]HLH tracer can result in serious overestimates of the immunological activity in the measurement of LH. Therefore [125I]HLHalpha should be separated from [125I]HLH prior to radioimmunoassay. Many of the fractionation methods commonly used (cellulose adsorption and short column gel filtration systems) are inadequate for this purpose. However, an adequate separation can be achieved by the use of high resolution gel filtration systems.

Biological Assay↗