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Biomedical subjects

H Funahashi

Publications and source records attributed to H Funahashi.

At least 145 records · Page 8Linked to original sources
PubMed1993

Effects of follicular fluid at fertilization in vitro on sperm penetration in pig oocytes.

The effects of porcine follicular fluid (PFF) on sperm penetration of pig oocytes and on prevention of polyspermy were examined and characteristics of spermatozoa exposed to PFF were determined. The addition of PFF at the level of 1 and 10% to the prefertilization and fertilization media decreased penetration rates and the mean number of spermatozoa in penetrated eggs regardless of the origin of PFF. In the presence of BSA, supplementation of 0.1% PFF to prefertilization and fertilization media and 1% PFF to prefertilization media did not decrease the penetration rates but did increase monospermic penetration to 54 and 68%, respectively. When PFF was added to prefertilization media, the number of spermatozoa binding to the zona and the percentage of acrosome-intact spermatozoa decreased with increased PFF concentration (from 43.1 +/- 2.8 and 73.1 +/- 4.9% to 7.2 +/- 1.3 and 15.7 +/- 15.4%, respectively). At the end of prefertilization incubation, sperm agglutination was observed and the degree depended on PFF concentration. Supplementation of fetal calf serum to prefertilization and fertilization media blocked the effects of PFF on sperm penetration and binding of spermatozoa to the zona. These results indicate that the prefertilization incubation of porcine spermatozoa in suitable concentrations of porcine follicular fluid will effectively reduce both the number of spermatozoa that attach to the surface of pig eggs and the incidence of polyspermy.

Acrosome↗
PubMed1993

Proliferating cell nuclear antigen immunostaining in breast cancer and its relation to prognosis.

Proliferating cell nuclear antigen (PCNA) appears in the cell nuclei during the late G1 to S phases of the cell cycle and is thought to be closely related to cellular proliferation. The authors have conducted an immunohistochemical study in order to investigate the tissue expression of PCNA and its clinical significance in breast cancer. Excluding cases with absolutely no positive cells on the section specimen, the mean value (%) for the PCNA labeling index (LI) was 30.4 in 187 cases of invasive ductal carcinoma. No correlations between PCNA LI and any clinicopathological factors such as tumor diameter and tumor stage were observed. Also, no significant correlation was observed with Ki-67 LI. A positive correlation was, however, observed with the tissue expression of c-erbB-2 protein. We divided 82 patients with stage II invasive ductal carcinoma into PCNA LI of < 10, PCNA LI of 10-50 and PCNA of > 50, and analyzed the specimens for any correlation with prognosis. The group with PCNA LI of > 50 had significantly poorer prognoses than the other groups. From the above, we concluded immunostaining for PCNA to be useful as a prognostic factor and as an indicator of the degree of malignancy in breast cancer.

Antigens, Neoplasm↗
PubMed1993

Our technique of parathyroid autotransplantation in operation for papillary thyroid carcinoma.

BACKGROUND: For papillary thyroid carcinoma, we have used our standard operative method consisting of total thyroidectomy, bilateral lymph node dissection, and parathyroid autotransplantation. Our special parathyroid transplantation method involves transplantation of the minced glands into the pectoralis major muscle. We investigated whether this method is useful for preserving parathyroid function. METHODS: In 17 patients who underwent our standard operation for papillary thyroid carcinoma, the intact parathyroid hormone (PTH) and serum calcium levels were determined for 4 weeks after the operation. RESULTS: In all cases the intact PTH levels fell below detectable limits immediately after operation and then recovered to the preoperative level about 2 weeks after the operation. The same recovery patterns were observed in two groups undergoing transplantation of two glands and three or four glands. CONCLUSIONS: These recovery patterns of intact PTH levels seem to indicate that the glands were grafted successfully and recovered their function. Our method is thought to be useful for preserving parathyroid function after surgery.

Calcium↗
PubMed1993

[Evaluation on our procedure for autotransplantation of parathyroid glands by the intact-PTH].

In thyroid cancer surgery since 1978 we have made parathyroidal autotransplantation which has been to resect the entire parathyroid, cut fine to mud and autotransplant into greater pectoral muscle. To evaluate our procedure and consider the mechanism of hypocalcemia and tetanus following surgery, we examined the intact-PTH recovered to about 80% of its preoperative value on 14-postoperative-days and thereafter remained almost constant. It remained below the sensitivity of measurement from immediately to 3-postoperative-days, and with our procedure, parathyroid was considered to be totally resected. Comparison between 2 (6 cases) and more autotransplanted glands (11 cases) revealed that the former showed slightly later functional recovery, but recovered to the almost some extent as the other 21-postoperative-days. Comparison between the group requiring calcium supplement therapy (12 cases) and the otherwise (5 cases) revealed almost the same course of recovery. Thus, our procedure seems to enable us to make satisfactory functional preservation and expect functional recovery by 2-gland autotransplantation at least. The supplement therapy was considered not to be detrimental to the take of autotransplanted glands and functional recovery, and no correlation was noted between the onset of tetanus and intact-PTH.

Adult↗
PubMed1992

C-erbB-2 protein in the sera of breast cancer patients.

The c-erbB-2 protein was measured in sera of patients with breast cancer or benign breast diseases to study the significance of this protein as a tumor marker. The mean value and positive rate for this protein (assuming 20U/ml as the cut-off value) were 11.8 U/ml (0%) in benign breast disease (n = 30), 11.8 U/ml (3.1%) in stage I/II primary breast cancer (n = 64), 38.2 U/ml (29.4%) in stage III/IV primary breast cancer (n = 17), 17.9 U/ml (33.3%) in locally recurrent breast cancer (n = 12), 298.4 U/ml (51.0%) in recurrent breast cancer with distant metastases (n = 51), and 12.9 U/ml (0%) in those with no evidence of recurrence (n = 57). Thus, the serum c-erbB-2 protein level was significantly higher in the distant metastatic group. In patients with distant metastases, there was a close association between expression of c-erbB-2 protein in the primary tumor and the serum c-erbB-2 protein level. On the basis of these results, serum c-erbB-2 protein was thought to be useful as a tumor marker for postoperative monitoring of breast cancer, especially in patients positive for expression of this protein in primary cancer tissue.

Breast Neoplasms↗
PubMed1992

Fertilization and early cleavage in vitro of ageing bovine oocytes after maturation in culture.

Bovine follicular oocytes cultured for 20 to 48 hours were inseminated with frozen thawed spermatozoa. Significantly higher penetration rates (94 to 100% vs 86 to 94%: P < 0.05) and proportions of polyspermy (35 to 78% vs 22 to 45%: P < 0.01) were obtained for oocytes cultured continuously for 24 hours with spermatozoa than for those separated from spermatozoa 8 hours after insemination. The most prominent effect of ageing of oocytes on early events of penetration was observed in the incidence of polyspermy rather than in the penetration rate and the proportion of pronuclear plus cleaved oocytes: the proportion of polyspermic oocytes significantly increased (P < 0.05) in oocytes inseminated after 28 to 48 hours of culture (36 to 78%) compared with those cultured for 20 to 24 hours (22 to 35%) for maturation. Culture experiments for early development of penetrated oocytes indicated that no significant differences were observed in the proportions of oocytes cleaved to the two- to four-cell stage 48 hours after insemination among those cultured for 20 to 40 hours for maturation. However, further cleavage to the four- to sixteen-cell stage 72 to 96 hours after insemination was greatly inhibited as ageing of oocytes proceeded from 28 hours in culture for maturation.

Journal Article↗
PubMed1992

Low frequency of rearrangements of the ret and trk proto-oncogenes in Japanese thyroid papillary carcinomas.

We investigated the frequency of rearrangements of the ret and trk proto-oncogenes in Japanese thyroid tumors. DNAs from 38 thyroid papillary carcinomas and 14 follicular adenomas were analyzed by Southern blotting. Rearrangements of the ret and trk proto-oncogenes were detected in one and two papillary carcinomas, respectively, but not in follicular adenomas. Analysis by a reverse transcriptase-polymerase chain reaction method showed that the ret rearrangement-positive tumor contained the PTC/retTPC chimeric transcript, which was reported to be found specifically in thyroid tumors and adenomatous goiter. We also found that rearranged mRNA of the trk proto-oncogene was expressed at high levels in one of two trk rearrangement-positive tumors. Our results indicated that the frequency of rearrangements of these proto-oncogenes in Japanese papillary carcinomas was much lower than that in Italian patients.

Adenoma↗
PubMed1992

ACTH increases expression of c-fos, c-jun and beta-actin genes in the dexamethasone-treated rat adrenals.

Our recent finding that ACTH increases c-fos mRNA in the adrenal gland of hypophysectomized rats indicates that the gene product FOS may play an important role(s) in mediating the action of ACTH. However, hypophysectomy employed in that study causes the disappearance of trophic hormones other than ACTH and may modify the effect of ACTH. Thus, in the present investigation, dexamethasone-treated rats were used. Since FOS functions only when it dimerizes with JUN (the product of c-jun gene), the changes in the levels of c-fos and c-jun mRNAs were studied together with that of beta-actin mRNA which is also affected by ACTH. Northern blot analysis was employed to determine the mRNA levels. It was demonstrated that ACTH increases the mRNAs coding c-fos and c-jun in the adrenal glands of dexamethasone-treated, ACTH-suppressed rats. The c-fos mRNA was not detectable before ACTH administration. After ACTH administration, the mRNA levels were transiently increased, the maximum level being observed at 30 min after ACTH. At 180 min post ACTH, the level returned to the unstimulated level. The mRNA coding c-jun was detectable before ACTH administration and it also increased rapidly after ACTH with maximal stimulation at 30 min. However, the mRNA level at 180 min post ACTH was still higher than the unstimulated level. The changes in beta-actin mRNA were approximately the same as those of c-jun mRNA. These results suggest that increased expression of c-fos, c-jun and beta-actin genes by ACTH may play an important role in mediating its action on the adrenals.

Actins↗
PubMed1992

Primary pigmented nodular adrenocortical disease (PPNAD): immunohistochemical and in situ hybridization analysis of steroidogenic enzymes in eight cases.

Primary pigmented nodular adrenocortical disease (PPNAD) is a rare but an interesting adrenocortical disorder associated with ACTH-independent hypercortisolism. We have studied eight cases of the adrenals with PPNAD by immunohistochemistry of all steroidogenic enzymes involved in cortisol biosynthesis (P-45scc, 3 beta-HSD, P-450c21, P-45017 alpha, and P-45011 beta) and also by performing in situ hybridization of P-45017 alpha in seven cases in order to localize the sites of specific steroidogenesis in this unique disorder. Immunoreactivity of all the enzymes examined was intense in almost all of the cells in adrenocortical nodules, especially the cells with abundant eosinophilic cytoplasm in all the cases examined. The internodular cortex, which demonstrated atrophy in five cases, normal appearance in two cases and hyperplasia in one case, was negative for the enzymes with an exception of 3 beta-HSD. Hybridization signals of P-45017 alpha were condensed over the nodules in in situ hybridization study, suggestive of an increased production of the enzyme itself in cortical cells of the nodules. These results may be consistent with autonomous cortisol production by the nodular cells and indicate that almost all of the cells in the nodules produce cortisol, which can also explain the presence of hypercortisolism despite small sizes of adrenals in PPNAD. Immunoreactivity of steroidogenic enzymes is observed in a small cluster of cortical cells with abundant eosinophilic cytoplasm located at the zona reticularis but not in adjacent non-nodular cortex, which may support an abnormal development of the zona reticularis as a possible pathogenesis of this disorder.

3-Hydroxysteroid Dehydrogenases↗
PubMed1991

First cleavage of enucleated rat eggs following transplantation of karyoplast removed from pronuclear eggs stored at 2 to 6 degrees C for various durations.

Pronuclear rat eggs were cultured for 24 to 48 hours at 37 degrees C after storage at 2 to 6 degrees C for 0 to 216 hours in medium. Very high proportions (89 to 97%) of eggs cleaved to the two-cell stage after storage for 0 to 48 hours. The proportion, however, decreased rapidly in eggs stored for 72 hours (60%), and eggs stored for more than 120 hours cleaved poorly. However, when male and female pronuclei from stored eggs were transplanted into enucleated fresh eggs, 92 to 100% of the fused eggs with karyoplast stored for 0 to 144 hours cleaved. Although the cleavage rate was reduced to 50% when karyoplast from eggs stored for 168 hours was transplanted, this reduction was not significant. Complete loss of cleaving ability was observed in fused eggs with the karyoplast stored for 216 hours. These results clearly indicate that the pronuclei of rat eggs can be stored for a longer period than the cytoplasm at low temperatures (2 to 6 degrees C).

Journal Article↗
PubMed1991

Developmental capacity of bovine oocytes collected from ovaries of individual heifers and fertilized in vitro.

Bovine follicular oocytes from individual heifers (n=49) were separately matured, fertilized with frozen-thawed spermatozoa and cultured with cumulus cells. Although there were great variations in the number (mean+/-SD=19.1+/-11.9) of oocytes collected from individual heifers and the percentages of the oocytes cleaved 48 hours after insemination (mean+/-SD=69.5+/-18.4) and developed to the morula stage 7 days after insemination (mean+/-SD=10.9+/-10.9), there were significant correlations between the numbers of oocytes collected and cleaved (the correlation coefficient: r=0.9336) or developed to morula stage (r=0.6633), indicating that oocytes from different heifers have the same developmental ability after in vitro fertilization. Ten morulae and 12 blastocysts which were obtained 7 and 8 days after insemination were transferred, one by one, to each uterine horn of 11 recipients. At Day 60 of pregnancy, 8 (80%) fetuses were identified in 4 (80%) of 5 recipients into which 10 embryos were transferred at Day -1 of synchrony. However, only 3 (25%) fetuses were identified in 2 (40%) of 6 recipients into which 12 embryos were transferred at Day 0 or +1 of synchrony.

Journal Article↗
PubMed1991

Dexamethasone-nonsuppressible cortisol in two cases with aldosterone-producing adenoma.

Forty-one patients with aldosterone-producing adenoma (APA) were subjected to a dexamethasone suppression test (DST) before surgery. Serum cortisol and urinary excretion of 17-hydroxycorticosteroids were suppressed by dexamethasone in 39 patients [DST(+)]. In two patients (cases A and B), they were not suppressed [DST(-)]. Clinical manifestations of the two DST(-) patients were similar to those of DST(+) patients. Hypertension, hypokalemia, high serum aldosterone levels, and suppressed PRA were found in all of the patients. The cut surfaces of the adenomas from all of the patients, including cases A and B, were golden yellow, which is typical of APA. However, atrophies of the adjacent normal tissues were evident exclusively in the two DST(-) patients. After removal of the affected adrenals, the serum cortisol level was suppressed by dexamethasone in one of the DST(-) patients (case B). These findings suggested autonomous cortisol production by APA. To evaluate whether cortisol could be produced from the adenoma tissue, the presence of several steroidogenic enzymes was studied by immunohistochemistry and mRNA analysis in the adenomas and the adjacent nonneoplastic adrenals from the 2 DST(-) and 5 DST(+) patients. Immunohistochemical analysis demonstrated that steroidogenic enzymes were expressed in APA tumor tissues from both DST(-) and DST(+) patients. In both groups, mRNAs coding steroidogenic enzymes were present not only in the nonneoplastic but also in the tumor tissues. Quantitative analysis of the mRNA levels revealed that in the adrenals from DST(+) patients, the mRNAs were more abundant in nonneoplastic tissue than in tumor tissue. However, in those from DST(-) cases, the mRNAs were much more abundant in the tumor tissues than in the nonneoplastic tissues. These results indicate that tumor cells of the two DST(-) patients autonomously synthesized not only aldosterone but also cortisol. The diameters of the tumors from the two DST(-) patients exceeded 3 cm, while those from other DST(+) patients were smaller. In patients with large APA, adrenal insufficiency should be anticipated upon removal of the tumor.

Adenoma↗
PubMed1991

[Study of hormone replacement therapy following total thyroidectomy in thyroid cancer--with special reference to the analysis of thyroid hormone peripheral effects, using indirect calorimetry].

Peripheral effects of thyroid hormones were examined using an indirect calorimetry in 18 patients with thyroid cancer before and after total thyroidectomy. Peripheral effects of exogenous thyroid hormones in TSH-suppression therapy after thyroidectomy were also studied. The subjects were maintained without hormone replacement for 3 weeks after total thyroidectomy. The ratio of resting energy expenditure to basal energy expenditure (REE/BEE) was determined before operation, before hormone replacement, and 1 and 5 weeks after the beginning of replacement, and the values were compared with changes in the blood thyroid hormone levels. Positive correlations were observed between the changes in endogenous thyroid hormone levels before and after total thyroidectomy and those in REE/BEE (free T3 vs. REE/BEE; r = 0.756, p less than 0.01), suggesting that evaluation of REE/BEE is clinically useful as an index of peripheral effects of thyroid hormones. Five weeks after the beginning of hormone replacement, T4 and free T4 were slightly range, and no enhancement of energy metabolism was noted. From these findings, the post-operative TSH suppression therapy carried out at our department is considered to be justifiable also from the viewpoint of energy metabolism.

Adult↗
PubMed1991

[Kinetics of PTH metabolism after total parathyroidectomy and autograft at forearm in patients with secondary hyperparathyroidism due to chronic renal failure].

Five patients with secondary hyperparathyroidism (2(0) HPT) due to chronic renal failure were investigated for PTH metabolism kinetics after total parathyroidectomy and autograft at the forearm (PTxG). We studied the kinetics of PTH for 24 hours after surgical treatment and gradually increased PTH levels when the parathyroid grafts became functional. It has been well known that the secreted intact PTH is peripherally cleaved in the region of reside 34 to yield amino- and carboxyl-terminal fragments. In this study, specific assays were used for intact PTH (INT-PTH), middle (HS-PTH, MM-PTH) and carboxyl-terminal (C-PTH) fragments. Half-lives were calculated from the mean values of all experiments by a two-compartment system; INT-PTH: 11.1 +/- 3.9 min, HS-PTH: 9.6 +/- 1.7 hrs., MM-PTH: 11.1 +/- 2.9 hrs., C-PTH: 10.1 +/- 1.2 hrs. (Mean +/- SD). The disappearance rate of biologically active intact PTH was rapid, whereas those of middle and carboxyl-terminal biologically inactive fragments were prolonged in the patients with chronic renal failure. The graft function was examined by PTH gradient between the venous samples in the grafted arm as compared to the non-grafted arm. PTH gradient of INT-PTH assay was over 1.5 one week following PTxG in all patients. This suggests that the intact PTH assay is a reliable predictor of clinical response to the graft function after PTxG.

Forearm↗