Biomedical subjects

T Monden

Publications and source records attributed to T Monden.

At least 91 records · Page 5Linked to original sources

PubMed1995

Transcriptional down-regulation by epidermal growth factor of TRH receptor mRNA in rat pituitary cells.

To gain insight into the mechanism underlying the epidermal growth factor (EGF)-induced changes in responsiveness to TRH and in the numbers of TRH receptors (TRH-Rs) in the pituitary, we investigated the transcriptional regulation by EGF of the TRH-R gene in GH4C1 cells. Northern blot analyses and binding studies revealed that EGF reduced both TRH binding and TRH-R mRNA levels in a dose- and time-dependent manner, while no significant changes were observed in beta-actin mRNA levels. Addition of actinomycin D caused an acute increase in the basal TRH-R mRNA level, and the rate of decrease of the TRH-R mRNA was identical in control and EGF-treated groups, suggesting that the stability of the TRH-R mRNA was not significantly affected in EGF-treated cells. Incubation with cycloheximide also induced an increase in the basal TRH-R mRNA level and completely reversed the EGF-induced reduction of TRH-R mRNA levels. Furthermore, a nuclear run-on assay demonstrated that the rate of transcription of the TRH-R gene was significantly inhibited in cells treated with EGF. We conclude that (1) EGF decreases the expression of the TRH-R mRNA largely by reducing its rate of transcription, and this action requires the synthesis of new proteins, and (2) inhibitors of protein and RNA synthesis cause a significant increase in the basal TRH-R mRNA level, suggesting that there may be a short-lived protein suppressing the TRH-R mRNA level in the pituitary.

Animals↗

PubMed1994

A frequent alteration of p53 gene in carcinoma in adenoma of colon.

In general, colorectal carcinoma is thought to originate mainly from adenoma, and this pathway is called the adenoma-carcinoma sequence. Carcinoma in adenoma is an appropriate model for analysis of this mechanism, because adenoma and carcinoma tissues coexist in the same polyp and the carcinoma is thought to have originated from the surrounding adenoma. Expression of the p53 protein was analyzed in 36 cases of carcinoma in adenoma in the colon by immunohistochemistry using an anti-human p53 monoclonal antibody (PAb1801). Alterations of the p53 gene were analyzed by the polymerase chain reaction for microanalysis of normal mucosa, adenoma, and carcinoma from histological slides. Mutations were assessed by the polymerase chain reaction-single strand conformation polymorphism analysis and identified by DNA sequencing in some cases. Loss of heterozygosity was studied by polymerase chain reaction-restriction fragment length polymorphism analysis. Positive staining for p53 was detected in three (8%) of 37 adenomas and 20 (53%) of 38 focal carcinomas. One (7%) of 15 adenomas with mild dysplasia, three (14%) of 22 adenomas with moderate dysplasia, and 16 (42%) of 38 focal carcinomas had a mutation in exon 5 through exon 8 of the p53 gene. As for allelic loss in the p53 gene locus, only one adenoma with moderate dysplasia had loss of heterozygosity, whereas six (40%) of 15 focal carcinomas had loss of heterozygosity. Of those tumors (3 of 37 adenomas and 20 of 38 focal carcinomas) that reacted with PAb1801, 78% (18 of 23) showed genetic alterations. Among 52 tumors which showed negative staining, five tumors had a p53 mutation and four of them were nonsense mutations. Putting all of these results together, 71% (24 of 34) of the cases underwent p53 gene and protein alterations during the conversion from adenoma to focal carcinoma. These data clearly indicate that genetic alterations of p53 are involved mainly in the malignant transformation from adenoma to focal carcinoma in colon carcinogenesis. In addition, some cases show heterogeneity of the p53 gene in carcinoma in adenoma of the colon. There may be other pathways than p53 responsible for malignant change in the colon.

Adenocarcinoma↗

PubMed1994

New development of transarterial immunoembolization (TIE) for therapy of hepatocellular carcinoma with intrahepatic metastases.

The prognosis of patients with multiple hepatocellular carcinoma (HCC) remains disappointing. In this study, we devised a new therapeutic modality for HCC consisting of transarterial immunoembolization (TIE) using OK-432 and fibrinogen and then analyzed the preliminary results. In the first series, we applied the treatment to 19 patients with advanced HCC who had proved to be insensitive to several previous conventional treatments. In all, 14 patients (74%) with unresected HCC have currently survived for between 2 and 16 months after TIE. The remaining 5 patients died at 17, 14, 8, 7, and 4 months after TIE. The serum levels of tumor markers decreased in all of the patients, and a marked reduction in tumor size was observed in six patients after TIE. A high fever occurred in all cases, and abdominal pain and loss of appetite were also observed after TIE. However, deterioration of liver function was negligible. After confirmation of the safety of this method, we started a second study series in which this TIE treatment was selected as the first choice. Six patients have been treated to date. All patients in this group underwent hepatic resection at 6-48 days following TIE. Histological examination of the resected specimens following TIE showed massive infiltration of mononuclear cells around tumor cell nests and lytic necrosis as well as coagulation necrosis of the main tumor and the intrahepatic metastases. In conclusion, our results indicate that TIE may be a safe and promising therapy for patients with HCC.

Adult↗

PubMed1994

Immunohistochemical study of p53 in gastric carcinoma.

The authors had previously reported that the accumulation of p53 is clearly demonstrable in microwave-fixed, paraffin-embedded sections of colorectal carcinomas. In the present paper, the authors performed an immunohistochemical study of p53 in gastric carcinomas prepared by microwave irradiation. Using a monoclonal antibody (PAb1801), nuclear p53 was detected in 22 of 45 (49%) cases of gastric carcinoma, and no staining for p53 was demonstrated in the adjacent normal epithelium, including areas of intestinal metaplasia. The incidence of p53 accumulation was not affected by clinical features, such as tumor stage, depth of invasion, and metastasis; however, positivity for p53 was significantly higher in intestinal-type carcinomas (56%) than in diffuse-type (27%) carcinomas. Furthermore, a large number of carcinoma cells expressed p53 in the intestinal-type, but only a few cells were positive for p53 in the diffuse-type. The results of the present immunohistochemical study of p53 accumulation in gastric carcinomas suggest that gene alterations of p53 are not rare in gastric carcinomas and may participate in the carcinogenesis of intestinal-type carcinomas of the stomach.

Gastric Mucosa↗

PubMed1994

Immunohistochemical analysis of p53 in gynecologic tumors.

Immunohistochemical staining for the p53 protein was performed in microwave-fixed, paraffin-embedded sections of normal, premalignant and malignant tissues of the female genital tract using a monoclonal antibody, PAb 1801. No staining was detected in normal and premalignant tissues, whereas nuclear staining of cancer cells was observed in 12 (22%) of 55 cervical squamous cell carcinomas, 4 (25%) of 16 cervical adenocarcinomas, 37 (42%) of 88 endometrial carcinomas, 23 (38%) of 60 ovarian adenocarcinomas, and 6 (100%) of 6 squamous cell carcinomas arising in dermoid cysts. Of interest, 1 of 7 endometrial cancers with concomitant atypical hyperplasia showed weak nuclear staining in a few atypical hyperplastic glands in addition to the cancerous lesions. Although staining was associated with cancers having a high histologic grade and serous papillary adenocarcinomas of the endometrium, it did not correlate with invasion, metastasis, or clinical stage. Comparison of the staining patterns with molecular analysis of mutations in the p53 gene showed the expected correlation of nuclear staining with missense mutations but not with nonsense mutations, which consistuted one third of all mutations found in this series. In addition, cytoplasmic staining did not predict mutation.

Cervix Uteri↗

PubMed1994

Pancreas cancer-associated antigen (PCAA) in medullary thyroid carcinoma.

Pancreas cancer-associated antigen (PCAA), primarily isolated from the ascites of pancreatic cancer (PC) patients, is strongly positive in PC, colon cancer and normal colonic mucosa. In immunohistochemical staining of tumor tissues with antibodies, medullary thyroid carcinoma (MTC) was no less strongly positive for PCAA than PC, and we studied its details. Antibodies to PCAA, calcitonin and CEA were used in the immunostaining of normal thyroid tissues and thyroid tissues from patients with adenomas, MTC, papillary carcinomas, and follicular carcinomas. The PCAA from the liver metastases of MTC was studied for molecular weight and antigenicity in comparison with the PCAA from the ascites of PC patients. Serum levels of PCAA were determined in MTC patients. Of 11 patients with MTC, PCAA, calcitonin and CEA were studied immunohistologically and positive in 10, 11 and 10 patients, respectively. The PCAA from the metastases had a molecular weight of about 700,000, and was immunochemically identical to that from the ascites of PC patients. Serum levels of PCAA were elevated in 4 of 6 MTC patients. The thyroid tissues from the MTC patients, familial or non-familial, were as strongly positive for PCAA as for calcitonin and CEA. It was antigenically identical to that of PC origin, and positive in the serum of MTC patients.

Adenocarcinoma, Follicular↗

PubMed1994

[Analysis of the TSH receptor gene structure in Graves' disease by a restriction fragment length polymorphism (RFLP) study].

We examined the structure of the TSH receptor gene in thyroid disorders, particularly in Graves' disease by a RFLP study with a cloned human TSH receptor cDNA as a probe. Southern blot analysis indicated that the TSH receptor gene is a single copy gene in the human genome. Although RFLPs were not detected in peripheral blood cells and thyroid tissues in patient with Graves' disease, significant RFLPs were found in 2 of 6 patients with adenoma. We conclude that there are no major abnormalities in the structure of the TSH receptor gene in patients with Graves' disease, and that DNA from thyroid adenomas can have large insertions or deletions.

DNA↗

PubMed1994

[Immunological and histological analyses of transarterial immuno-embolization therapy (TIE) in operable patients with hepatocellular carcinoma].

Immunological and histological analyses were performed on 14 patients with hepatocellular carcinoma treated by transcatheter immunoembolization (TIE) and subsequently by hepatic resection. They were compared with the cases treated by transcatheter arterial embolization (TAE). Exceptionally high plasma levels of inflammatory cytokines, such as IL-6 and IL-8, were noted 3 hours after TIE insults in the majority of the cases. On the contrary, exceptionally high levels of TNF-alpha were also observed in some cases of TIE treatment. In addition, light microscopically, the lytic necrosis of the tumor and massive infiltration of mononuclear cells were the histological characteristics of this treatment. Interestingly, the population of the infiltrates has altered after TIE treatment. It thus consisted mainly of neutrophils in early phase, subsequently of the mixture of lymphocytes, eosinophils, and plasma cells, and finally of lymphocytes. These results may suggest that certain inflammatory responses caused by TIE may play important roles in this new therapeutic modality.

Carcinoma, Hepatocellular↗

PubMed1993

Pituitary adenomas of patients with acromegaly express thyrotropin-releasing hormone receptor messenger RNA: cloning and functional expression of the human thyrotropin-releasing hormone receptor gene.

We cloned a human thyrotropin-releasing hormone receptor (TRH-R) gene and its pituitary cDNA, and examined whether TRH-R mRNA may be expressed and function in the GH-secreting pituitary adenomas of patients with acromegaly. The human TRH-R consists of 398 amino acids and is a member of the G protein-coupled receptor family. The gene has a single intron in the coding sequence. An addition of TRH caused an obvious increase in intracellular calcium concentration in chinese hamster ovary cells expressing this receptor. Although highly homologous to the rat and mouse TRH-Rs, the human TRH-R has several differences in the carboxyl-terminal structure. The sequence and functioning analyses showed the same expression of TRH-R mRNA in the pituitary adenomas as that in the normal pituitaries, indicating that 1) some adenomas of patients with acromegaly express an intact and functional TRH-R, and that 2) the paradoxical response of GH secretion to TRH found in the patients might be a direct effect of TRH on TRH-R expressed in the adenoma.

Acromegaly↗

PubMed1993

Assignment of human thyrotropin-releasing hormone (TRH) receptor gene to chromosome 8.

The human gene encoding thyrotropin-releasing hormone receptor was assigned to chromosome 8, using human-Chinese hamster ovary somatic cell hybrids, analyzed by Southern hybridizations. Hybridization was carried out with a 32P-labeled fragment of the human TRH-R genomic DNA. Hybridization of this probe to a human specific 10.5-kb DNA fragment of EcoRI-digested WBC DNA was used to localize the human TRH-R gene. No hybridization, by contrast, was seen with this probe and hamster DNA after EcoRI treatment. Results from 18 somatic cell hybrids corroborated unequivocally that the human TRH-R gene can be assigned to human chromosome 8.

Animals↗

PubMed1993

Significance of p53 expression as a prognostic factor in oesophageal squamous cell carcinoma.

The tumour suppressor gene product p53 is believed to play an important role in the progression of human malignant tumours through mutation and over-expression. Using a microwave oven heating method, we have detected over-expression of p53 in buffered-formalin fixed, paraffin-embedded sections of oesophageal carcinomas immunohistochemically and examined the relationship between the p53 over-expression and postoperative survival. Employing a monoclonal antibody (pAb1801), nuclear p53 was detected in 56 of 105 (53%) tumour specimens. Homogeneous, heterogeneous, and focal immunostaining patterns were noted. No immunostaining was found in adjacent benign tissues. The results in buffered-formalin fixed sections were similar to those in the frozen sections. The cumulative survival rate of patients with p53 expression was significantly lower than that of the patients without expression (P < 0.05), even though there were no significant differences between the clinicopathological features of the two groups. The results indicate that the nuclear accumulation of p53 might be an independent prognostic factor in patients with oesophageal squamous cell carcinomas.

Adult↗

PubMed1993

Augmentation of antitumor immunity in regional lymph nodes by local immunotherapy.

We have previously reported that the antitumor effect of OK-432, a streptococcal preparation, is markedly augmented when injected intratumorally together with fibrinogen (OK-432/fbg) [1]. In order to elucidate the effects of this immunotherapy on regional lymph nodes (RLN), we carried out both morphological and functional analyses of the RLN from colonic cancer patients treated with OK-432/fbg. Computer-aided morphometry revealed that the maximal cross-sectional areas and the broadest diameters of the RLN were significantly greater (p < 0.01) in patients who had undergone local immunotherapy than in patients who had not. The component structures of RLN, such as sinus, follicle and paracortex, were all enlarged in the OK-432/fbg-treated patients, and necrosis of metastatic tumors was observed. RLN lymphocytes recovered from OK-432/fbg treated patients showed elevated reactivity to phytohemagglutinin (PHA) and the stimulation index was clearly higher than that of control patients. Flow cytometric analysis revealed a predominance of T-cells, especially CD4 subsets, and higher positivity for both CD25 and HLA-DR. Furthermore, RLN lymphocytes killed more effectively K562 and Daudi cells in the patients who had had immunotherapy. These results suggest that the effect of local immunotherapy with OK-432/fbg is not restricted to the site of injection but extends to the lymph nodes, and contributes to tumor regression through the augmentation of cellular immunity.

Aged↗

PubMed1993

The effect of local immunotherapy for breast cancer using a mixture of OK-432 and fibrinogen supplemented with activated macrophages.

OK-432 is an immunomodulatory agent prepared from a strain of Streptococcus pyogenes. We have previously reported that intratumoral injection of a mixture of OK-432 and fibrinogen (hereinafter referred to as OK/fbg) is very effective in the local immunotherapy for colorectal cancer. However, we found that the intratumoral injection of OK/fbg into tumor tissues of breast cancers did not always induce a strong antitumor effect. With conventional OK/fbg treatment, tumor necrosis observed in breast cancer tumors was significantly less than that in colorectal cancer tumors; the formation of fibrin meshwork and macrophage infiltration, in particular, were poor. In this study, the OK/fbg mixture was supplemented with activated macrophages for local immunotherapy of breast cancers. Macrophages were prepared from peripheral blood of breast cancer patients and activated with 0.05 mg/ml of OK-432. Between 2-7 days before operation, a single intratumoral injection of the above mixtures was done. The addition of activated macrophages to the OK/fbg mixture resulted in marked degrees of fibrin meshwork formation, macrophage infiltration and cancer cell necrosis. These findings suggest that the recruitment of macrophages in tumor stroma and their activation are necessary for sufficient induction of antitumor immunity, and supplementation of activated macrophages at the site of immune reaction may be an alternative method for reinforcement of the antitumor effect of local immunotherapy.

Breast Neoplasms↗

PubMed1993

A cancer-reactive human monoclonal antibody derived from a colonic cancer patient treated with local immunotherapy.

A human monoclonal antibody, YJ-37 (IgM) was generated through the fusion of human B lymphoblastoid cell line HO-323 with the regional lymph node lymphocytes from a colonic cancer patient who was treated with a local immunotherapy. This antibody was purified and conjugated with biotin, after which direct immunohistochemical staining was performed. The results revealed that YJ-37 selectively reacted with colonic cancer (7/19), gastric cancer (3/6), endometrial cancer (1/2) and colonic adenoma (7/13), but not with normal epithelia. Membrane immunofluorescence and FACS analysis also showed that YJ-37 bound to tumor cell surfaces. Furthermore, the chemical structure of the antigen defined by YJ-37 was analyzed by means of thin-layer chromatography immunostaining and ELISA. The results indicated that YJ-37 reacted with sialylated lacto-series carbohydrate chains, which have been reported to accumulate in cancer cells.

Adenoma↗

PubMed1993

Alteration by prolonged oral administration of a thyrotropin-releasing hormone analog, TA-0910, of the pituitary-thyroid axis in subjects with brain stroke.

We evaluated whether prolonged oral administration of a novel analog of thyrotropin-releasing hormone (TRH), TA-0910, may change the pituitary-thyroid axis in human subjects with brain stroke. The subjects were given one oral dose of 2.5-20 mg of TA-0910 daily for 8 weeks, and then blood levels of TA-0910, thyroid hormones and thyrotropin (TSH) were measured. Plasma levels of TA-0910 were elevated with increasing doses. After administration of TA-0910, the basal levels of thyroid hormones and TSH had a tendency to increase and decrease, respectively, but those changes remained in the normal range. The secretion of TSH in response to TRH was not significantly affected by TA-0910 administration. No significant changes in other pituitary hormones were observed after TA-0910 administration. The present data indicate that prolonged oral administration of TA-0910 did not significantly alter the pituitary-thyroid axis in subjects with brain stroke.

Administration, Oral↗

PubMed1993

Preparation of a human monoclonal antibody derived from cervical lymph nodes of a patient with anaplastic carcinoma of the thyroid.

An anaplastic carcinoma cell line, KOA-2, was established from a 59-year-old patient with anaplastic carcinoma of the thyroid who died only one month after diagnosis in association with rapid progress of her disease. A human monoclonal antibody, 3C5 (IgM), was generated by a fusion of human B lymphoblastoid cell line, HO-323, with lymphocytes from cervical lymph nodes of the patient, who had been treated with local immunotherapy. Immunocytostaining demonstrated that 3C5 reacted with KOA-2, and membrane immunofluorescence demonstrated that 3C5 reacted with cell membranes. Immunohistochemical staining studies demonstrated selective reaction of 3C5 with three malignant tumors, anaplastic carcinoma of the thyroid (2/4 sampled tested), papillary carcinoma of the thyroid (8/12), and breast cancer (2/6), but no reaction with specimens of benign thyroid disease or normal thyroid gland was demonstrated. The monoclonal antibody and the cell line established from one patient have potential use in the analysis of carcinogenesis and applications to therapy of anaplastic carcinoma of the thyroid.

Antibodies, Monoclonal↗

PubMed1993

Edema due to altered sweating function.

We report a case with idiopathic edema accompanied with excessive sweating on bathing. The subject is a 26 year-old male. He was admitted to our hospital because of pretibial edema. He complained of excessive sweating only after daily bathing. Sweating on daily bathing reduced pretibial edema after long-term standing. Daily urine volume decreased by about 200 to 500 mL/day on bathing everyday. Plasma renin, aldosterone and bradykinin levels increased. Subcutaneous injection of pilocarpine increased sweating around the injection site. The cessation of daily bathing restored plasma renin, aldosterone and bradykinin levels; urine volumes increased. However, pretibial edema appeared and body weight increased by 8.5 Kg. Serum triiodothyronine (T3), thyroxine (T4) levels and basal metabolic rate (BMR) decreased. Circulating plasma volume increased with the cessation of bathing. Alterations in the autonomic nervous system may be involved in the appearance of general edema through increased plasma volume due to altered sweating function.

Adult↗

PubMed1992

Differential regulation of thyrotropin-releasing hormone receptor mRNA levels by thyroid hormone in vivo and in vitro (GH3 cells).

We studied the effect of thyroid status on thyrotropin-releasing hormone receptor (TRH-R) mRNA levels both in vivo and in vitro (GH3 cells) using a cloned rat TRH-R cDNA by RT-PCR. Experimental hypothyroid rats were produced by total thyroidectomy and were then killed 7 days after the operation. TRH receptor binding in the anterior pituitary and serum TSH level were elevated approximately 2-fold and 8-fold, respectively, in 7 day thyroidectomized rats. TRH-R mRNA levels in hypothyroid rats were also increased significantly compared with those of normal rats. In GH3 cells, however, no significant change of TRH-R mRNA level was observed between cultures treated with triiodothyronine (T3, 10(-9) and 10(-7) M) and the untreated group. The present data indicate that 1) the in vivo effects of thyroid status on TRH-R mRNA levels differ from the in vitro one, and that 2) the down regulation of TRH-R binding by thyroid hormone in GH3 cells may be mediated by translational or post-translational mechanisms.

Amino Acid Sequence↗