Biomedical subjects

T Kameya

Publications and source records attributed to T Kameya.

At least 37 records · Page 2Linked to original sources

PubMed2000

Simultaneous suppression of cdc2 and cdk2 activities induces neuronal differentiation of PC12 cells.

The involvement of cdc2 and cdk2 during neuronal differentiation in rat pheochromocytoma PC12 cells was examined. When PC12 cells were cultured with nerve growth factor (NGF), expression of cdc2 decreased significantly after day 5, while expression of cdk2 decreased gradually after day 7. Cells overexpressing cdc2 or cdk2 were resistant to NGF-induced differentiation and growth suppression, and maintained high cdc2 or cdk2 kinase activity, respectively, during NGF treatment. In contrast, the NGF-treated parental cells showed a marked decline in these kinase activities after day 3. When PC12 cells were treated with specific inhibitors of cdc2/cdk2 (butyrolactone-I, olomoucin), they showed marked neurite extension and up-regulation of microtubule-associated protein 2 expression. In addition, treatment with mixtures of antisense oligonucleotides for cdc2 and cdk2 resulted in down-regulation of both cdc2 and cdk2 kinase activities as well as significant neurite outgrowth and up-regulation of microtubule-associated protein 2 expression. However, neurite outgrowth was not observed in cells treated with either single antisense oligonucleotide, or antisense cdc2 + cdk4 or cdk2 + cdk4 oligonucleotide mixtures. These results suggest that simultaneous down-regulation of cdc2 and cdk2 activity is sufficient and necessary for neuronal differentiation in PC12 cells.

Animals↗

PubMed2000

Involvement of cyclins in cell proliferation and their clinical implications in soft tissue smooth muscle tumors.

Expression of cyclins A and E and cyclin-dependent kinase 2 (cdk2) was examined immunohistochemically in 55 cases of soft tissue smooth muscle tumors, including vascular leiomyoma, and compared to expression of Ki-67 and proliferating cell nuclear antigen. Cyclin A was expressed in 70% of the leiomyoma cases, but with much lower labeling indexes than in leiomyosarcoma. Cyclin E was expressed exclusively in leiomyosarcoma. Although the differences of cyclin A- and cyclin E-labeling indexes between leiomyoma and leiomyosarcoma were statistically significant, no significant differences were found in the other markers. Furthermore, cyclin A- and/or E-positivity predicted a poor prognosis in recurrence- or metastasis-free survivals and overall survival. Immunoblotting revealed that cyclins A and E were expressed, in complex with cdk2, exclusively in tumors. In addition, not only leiomyosarcoma, but also leiomyoma specimens that exhibited negligible levels of complex expression, manifested detectable cdk2 activity. These results suggest 1) up-regulation of active cyclin A/cdk2 expression and associated kinase activity is critical for unrestrained cell proliferation; 2) cyclin E/cdk2 complexes may play a crucial role in leiomyosarcoma; 3) immunohistochemical detection of cyclins can be a more reliable tool for differential diagnosis between leiomyoma versus leiomyosarcoma than that of Ki-67 or proliferating cell nuclear antigen, and be a possible prognostic indicator.

Adult↗

PubMed2000

Semiquantitative immunoblot analysis of nm23-H1 and -H2 isoforms in adenocarcinomas of the lung: prognostic significance.

Total amounts of nm23 protein and relative levels of H1 and H2 isoforms were studied in 27 fresh-frozen samples of pulmonary adenocarcinoma and adjacent non-neoplastic tissues that were obtained at surgery. Semiquantitative immunoblotting with a monoclonal antibody (Pan-242) against nm23 protein demonstrated both isoforms, recognized as 20.5 kDa for H1 and 18.5 kDa for H2, to be present in all cases. Both H1 and H2 levels in neoplastic tissues were higher than in the corresponding non-neoplastic samples. Expression of H2 was usually greater than of H1. The H2/H1 ratio varied from 1.9 to 14.1 (mean value 5.2) in non-neoplastic tissues and 1.0-5.9 (mean value 2.5) in neoplastic tissues, although this ratio did not correlate with any prognostic factor like tumor size, nodal status or distant metastasis (TNM tumor stage). H1 and H2 levels were significantly lower (mean values 4.3 and 2.4) in well-differentiated than in moderately and poorly differentiated adenocarcinomas (8.3 and 3.0) (P < 0.03 and P < 0.05, respectively). These data indicate that H1 and H2 isoform levels correlate with histological differentiation, but not the metastatic potential or stage of pulmonary adenocarcinoma.

Adenocarcinoma↗

PubMed2000

Pulmonary adenocarcinoma with heterotopic bone formation.

Adenocarcinoma of the lung is a common malignancy. Frequently, this tumor can cause calcification within a primary tumor. However, an extremely rare occurrence in lung carcinomas is ossification within a primary tumor, and to our knowledge only three cases have been reported. We report a case of pulmonary adenocarcinoma with ossification, and discuss the pathogenesis of intratumoral ossification with a review of the literature.

Adenocarcinoma↗

PubMed2000

Cardiac dendritic cells and acute myocarditis in the human heart.

Cardiac dendritic cells are considered to play an important role in the immunoresponse of the heart. However, it is unclear whether these cells occur in human myocarditis and whether they function in similar ways to those in rats. Cardiac samples were obtained from 22 autopsied patients with myocarditis, and compared with 20 age-and sex-matched controls. Formalin-fixed hearts were immunostained by the LSAB method. Cardiac dendritic cells were detectable even in the control hearts (1.5 cells/high power field (HPF)). In the acute phase of myocarditis, the number of cardiac dendritic cells increased up to 12.6 cells/HPF (p<0.001). In the subacute phase of myocarditis, T cells (36.6 cells/HPF) and HLA-DR+ cells (10.2 cells/HPF) continued to infiltrate the periphery of the inflammatory lesions, but they had no expression without inflammation. In this study, cardiac dendritic cells were reactive for HLA-DR, but negative for CD68, and were characteristically large monocytes with long, slender, dendritic processes. Accordingly, they were clearly distinguishable from macrophages. In the human heart, cardiac dendritic cells may be recruited in the acute phase of myocarditis, and seem to play an important role in the succeeding immunoresponse.

Acute Disease↗

PubMed2000

Thymic hyperplasia as a source of ectopic ACTH production.

A 26 year-old man with suspected Cushing's disease underwent transsphenoidal exploration of the pituitary without any evidence of microadenoma or hyperplasia. Progressive hypercortisolism necessitated bilateral adrenalectomy. Postoperatively, skin pigmentation gradually developed with a marked elevation of plasma ACTH levels, and CT scanning uncovered a thymic mass. Following removal of the thymic mass, skin pigmentation disappeared and plasma ACTH levels fell to normal. The excised mass was found to be a benign thymic hyperplasia without epithelial or carcinoid tumor cells. However, gel chromatography showed that the thymic tissue extract contained high ACTH content comparable to that of ectopic ACTH-producing tumors with a major component corresponding to ACTH(1-39). Northern blot analysis and in situ hybridization revealed the expression of proopiomelanocortin transcripts in lymphocytes of thymic hyperplasia. This report suggests that lymphocytes in thymic hyperplasia are the most likely site of deregulated ACTH expression causing ectopic ACTH syndrome.

ACTH Syndrome, Ectopic↗

PubMed2000

Relationship between p53 gene mutation and protein expression: clinical significance in transitional cell carcinoma of the bladder.

Although the mutated p53 gene has been postulated to induce immunohistochemically-detectable p53 protein, reports regarding the relationship between p53 mutation and p53 protein expression have been contradictory. This study investigated the relationship between p53 mutations and p53 expression and their clinical significance for patients with transitional cell carcinoma of the bladder. Eighty-seven transitional cell carcinoma of the bladder were analyzed by immunohistochemistry (IHC) for p53 nuclear accumulation, and the results compared to mutations detected in the p53 gene evaluated by polymerase chain reaction single-strand conformation polymorphism (SSCP) and DNA sequence analysis. By p53 IHC analysis, positive p53 staining was observed in 50 (57.5%) of the 87 tumors. The specificity of IHC, defined as a percentage of IHC negative (<20%) tumors among tumors without mutation, was 94.6%. Despite the good concordance between p53 mutation and p53 protein expression (p<0.0001), 48.0% (24/50) of the tumors showed p53 overexpression without mutation, and 2 (5.4%) tumors with mutation showed no p53 immunoreactivity. Patients with higher grade (grade 3), stage (stages pT2-4), and p53 mutations had a poorer prognosis by Kaplan-Meier survival analysis. A Cox univariate analysis found that grading (hazard ratio 3.139; p=0.002), staging (hazard ratio 3.832; p=0.0005) and p53 mutation (hazard ratio 2.498; p=0.013) were significant variables in these patients, but no variable was independently associated with an increased survival of bladder carcinoma by multivariate analysis. We found that a 20% cut-off level of p53 overexpression showed the highest correlation with prognosis and p53 mutation, however, p53 overexpression and mutation were not superior to staging as prognostic markers. These data suggest that careful assessment of the TNM staging system remains the most reliable predictive indicator of survival for patients with transitional cell carcinoma of the bladder.

Adult↗

PubMed2000

Induction of neutrophil death resembling neither apoptosis nor necrosis by ONO-AE-248, a selective agonist for PGE2 receptor subtype 3.

An increase of intracellular cAMP mediated by prostaglandin E2 (PGE2) has been shown to delay spontaneous apoptosis of neutrophils. It has been demonstrated that a selective agonist for PGE2 receptor subtype 3 (the EP3 receptor) is capable of decreasing cAMP and stimulating phosphoinositide turnover in various types of cells. We investigated the effect of a selective EP3 receptor agonist, ONO-AE-248, on neutrophil viability. ONO-AE-248 rapidly caused a unique form of neutrophil death. The agonist primarily induced morphological changes of the nucleus, including fusion of the lobules, decreased compactness of the chromatin, and blebbing and rupture of the nuclear membrane. This was followed by an increase of plasma membrane permeability and cell lysis. During these processes, neither apoptotic changes such as nuclear condensation, DNA fragmentation, and expression of phospholipid phosphatidylserine on the plasma membrane nor necrotic changes such as chromatin clumping and organelle destruction were apparent in the treated neutrophils. The fatal effect of the agonist night be specific for neutrophils because it failed to promote the rapid death of other types of cells. Although activation of neutrophils by ONO-AE-248 was not evident, experiments using metabolic inhibitors demonstrated that the agonist caused neutrophil death via the activation of protein kinase C in the presence of intracellular ATP. These findings indicated that EP3 receptor-mediated signals might promote a novel form of neutrophil death, which differs from typical apoptosis or necrosis.

Apoptosis↗

PubMed2000

[A survey of current nutrition therapy for the ALS patients in Japanese national sanatoriums].

To evaluate the current nutrition therapy for the patients of amyotrophic lateral sclerosis (ALS) in Japan, we sent questionnaires to 53 Japanese national sanatoriums (hospitals) and were able to collect replies from 42 institutes. In these replies, information on 466 patients (262 inpatients and 204 outpatients) was reported, accounting for about 10% of all Japanese ALS patients. Patients who ate orally were more frequent (p < 0.001, chi 2 test) in the outpatients (76.0%) than in the inpatients (39.7%). When the subjects were limited to non-orally nourished cases, gastrostomy was more frequently performed (p < 0.05, chi 2 test) in the outpatients (49%) than in the inpatients (29.1%), but no significant differences were observed between the above two groups as to the quantity of daily caloric intake; the mode was 1000-1200 Cal. About a quarter of non-orally nourished patients received supplemental sodium chloride, and fewer were supplemented copper and/or zinc. The consent of the patients as well as the clinical findings weighed heavily in the determination of the feeding route. Most physicians were disinclined to encourage tube feeding in the early stage of dysphagia, but were supportive of the operation of percutaneous endoscopic gastrostomy when non-oral nutrition therapy became necessary. Although the efficacy of vitamins for ALS has not been proved, 45 of 58 physicians agreed to subscribe vitamins to their patients, especially vitamin B12, E and C.

Amyotrophic Lateral Sclerosis↗

PubMed1999

A novel apoptotic cascade mediated by CDK4 in rat pheochromocytoma PC12 cells.

Apoptosis induced by serum withdrawal in pheochromocytoma PC12 cells is promoted by overexpression of cyclin-dependent kinase 4 (CDK4). We compared CDK4-promoted apoptosis with that induced by serum withdrawal alone in PC12 cells. Protein synthesis inhibitors did not prevent apoptosis in parental cells, but prevented the promotion of apoptosis by CDK4 overexpression. Nerve growth factor, basic-fibroblast growth factor, and Bcl-2 proteins protected both parental and CDK4-overexpressing cells from apoptosis. However, insulin-like growth factor-I and Bcl-X(L) protein only partially inhibited apoptosis in the CDK4-overexpressing cells. Bcl-2 or Bcl-X(L) had no significant effect on CDK4 kinase activity in both cell lines. These results suggest a novel CDK4-mediated apoptotic cascade which is normally restrained, but which is activated by CDK4 overexpression. This apoptotic cascade should eventually converge with the cascade induced by serum withdrawal in normal PC12 cells. We discuss the interactions among these apoptotic cascades and the points where anti-apoptotic agents act.

Animals↗

PubMed1999

Which technique, circular stapled anastomosis or double stapling anastomosis, provides the optimal size and shape of rectal anastomotic opening?

BACKGROUND: Two types of end-to-end colorectal anastomosis using a circular stapler, circular stapled anastomosis(CSA) and double stapling anastomosis(DSA), have become the standard technique. The possibility of anastomotic openings of the DSA being smaller and somewhat distorted, because of some portion of the stapled rectal stump not being included in the anastomotic opening, was examined. METHODS: Anastomotic openings created by CSA and DSA were photographed through a sigmoidoscope, and the maximal and minimal diameters and the areas of the anastomotic openings were measured and calculated with an image analyzer. They were examined in the swine rectum immediately after surgery and were also examined in the randomized clinical cases 3 or more months after surgery. RESULTS: It was found that larger anastomotic openings could be created as the diameter of the anastomosed intestine and the cartridge size became wider in both methods immediately after surgery. The shape of anastomotic opening created by DSA was observed to be more oval than round immediately after surgery. The anastomotic stenosis occurred in a small number of patients in both methods within 3 months after surgery. The area of the anastomotic opening created by CSA was about 25% larger than that by DSA immediately after surgery and about 30% larger 3 or more months after surgery. CONCLUSION: CSA provides a larger anastomotic opening than DSA. Anastomotic stenosis occurred in a small number of patients in both methods.

Aged↗

PubMed1999

Human cytomegalovirus infection in foci of Langerhans cell histiocytosis.

Langerhans cell histiocytosis (LCH) has been thought to be a disorder of immune regulation, and increasingly, evidence showing that the tissue damage in LCH involves lymphokines and pro-inflammatory cytokines is reported. We detected human cytomegalovirus (HCMV)-DNA in LCH cells in the foci of LCH lesions by immunohistochemistry, in situ hybridization and PCR. HCMV was detected in the nuclei and/or cytoplasm of LCH cells in 9 of 27 LCH cases by immunostaining. HCMV was probably an early antigen. In situ hybridization revealed signals for HCMV-DNA only in the nuclei of LCH cells in 10 of the 27 LCH cases. PCR analysis was performed in 20 of the LCH cases, and HCMV-DNA was detected in 7 of these. All 7 positive cases were also positive for HCMV by ISH and IHC. These findings suggested that early phase infection or reactivation of HCMV occurred in the LCH lesions. HCMV infection may be accompanied by impaired cytokine production. Our study also suggested a relationship between HCMV infection and expression of TNFalpha. In tissues affected by LCH, dermatopathic lymphadenopathy or malignant fibrous histiocytoma and in normal tissues no signals for Epstein-Barr virus-RNA were detected. These findings suggest that in some cases LCH is associated with HCMV infection.

Adolescent↗

PubMed1999

Tumor extension and cell proliferation in adenocarcinomas of the lung.

To elucidate the mechanism of tumor extension in human pulmonary adenocarcinoma, we immunohistochemically investigated the expression of cell cycle regulator proteins in 54 small adenocarcinomas less than 3 cm in diameter. The Ki-67-labeling index was significantly higher in the periphery of the tumor nodule than in the center. This proliferative potential correlated well with coexpression of cdk2 and cyclin A. p27, one of the cdk inhibitors, was highly expressed in normal bronchial epithelial cells. Peripherally located tumor cells expressed p27 at an intermediate level, but at a higher frequency and level than those in the center. Expression of p21 was also predominant in the periphery. Furthermore, the expression patterns of p21 and p27 were reciprocal. In vitro kinase assays further demonstrated higher cdk2 kinase activity in the periphery. These results suggest that: (i) within an emerging extension made up of peripherally located tumor cells, their high proliferative potential gradually wanes as their relative topographical position becomes more central in the expanding tumor; (ii) peripherally located tumor cells maintain their proliferative potential by higher cyclin A-cdk2 complex activity; and (iii) intermediate expression of p21/p27 in the peripherally located cells promotes higher cyclin A-cdk2 kinase activity, whereas high p21/p27 expression in nonneoplastic cells inhibits kinase activity.

Adenocarcinoma↗

PubMed1999

Leptomeningeal melanomatosis with multiple cutaneous pigmented nevi: tumor cell proliferation and malignant transformation in an autopsy case.

We experienced a rare case of leptomeningeal melanomatosis. The proliferative activity and nuclear accumulation of p53 in this tumor were examined, since the relationship between this tumor type and growth has not yet been elucidated. A 33-year-old Japanese man was shown to have leptomeningeal melanomatosis with multiple cutaneous pigmented nevi. The autopsy findings showed the presence not only of benign diffuse melanosis of the leptomeninges but also of leptomeningeal melanomatosis in the subarachnoid space and brain parenchyma. In the brain parenchyma, the direct invasion of tumor cells from the subarachnoid space and Virchow-Robin spaces filled with melanoma cells were observed. Multiple hemorrhagic areas invaded by melanoma cells were also present. Immunohistochemical staining with a monoclonal antibody to melanoma cells showed positivity in the tumor cells. Proliferation analysis using the MIB-1 antibody demonstrated that the labeling index of tumor cells invading brain parenchyma (2.54%) was higher than that in other lesions of the inner (0.89%) and outer layer (0.76%) of the subarachnoid space. Nuclear accumulation of p53 protein was rarely seen in the tumor cells. We reported a case of leptomeningeal melanomatosis. Higher proliferative activity was found in invading cells of the brain parenchyma. Malignant transformation of the tumor did not appear to be associated with p53 gene mutation.

Adult↗

PubMed1999

Significance of growth hormone-releasing hormone receptor mRNA in non-neoplastic pituitary and pituitary adenomas: a study by RT-PCR and in situ hybridization.

This study examined the expression of human growth hormone-releasing hormone receptor (GHRH-R) mRNA in both non-neoplastic pituitary tissues and pituitary adenomas by reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH). RT-PCR analysis showed that all of the non-neoplastic pituitaries and all GH-producing adenomas, one prolactinoma and one third of the non-functioning adenomas expressed GHRH-R mRNA. ISH demonstrated that all of GH-producing adenomas and two prolactinomas expressed GHRH-R mRNA. The expression of GHRH-R mRNA in GH-producing adenomas was greater than that in the other adenomas by RT-PCR and ISH. GHRH-R mRNA detected by ISH was observed only in GH cells from the pituitary gland of a young girl. In pituitary adenomas, a diffuse signal was observed in the cytoplasm of all of the GH-producing adenomas and in two prolactinomas. Expression of GHRH-R mRNA was not seen in normal prolactin cells, or in any adenomas other than GH-producing adenomas and a few prolactinomas. These results suggest that GHRH-R mRNA plays a role mainly in the function of GH-producing adenomas but may also play a role in function of some prolactinomas.

Adenoma↗

PubMed1999

Morphological characterization and subtyping of silent somatotroph adenomas.

GH-producing adenomas clinically are endocrine-active tumors accompanied with acromegaly in most instances. However, GH-producing adenomas apparently unassociated with acromegaly, or so-called silent somatotroph adenomas (SSA), have recently been reported but rarely. The reported cases are characterized by normal or slightly elevated serum levels of GH but without acromegaly. Tumor cells contain moderate, trace or no GH immunoreactivity. We experienced 7 cases of SSA which were not always similar in morphology and pathogenetic mechanism. They could be further divided into the following 3 subtypes. Subtype 1 (N = 2): a moderate number of cells were immunopositive for GH, and GH mRNA was also expressed in moderate or numerous cells. Densely granulated cells were noted. It is assumed that inhibition of hormone release into circulation. Subtype 2 (N = 3): a small number of cells were immunopositive for GH, while GH mRNA was expressed in numerous tumor cells. They were sparsely granulated cells containing fibrous bodies. These findings suggest that posttranslational processing of the gene product may be defective. Subtype 3 (N = 2): Only a scattered number of cells were immunopositive for GH and GH mRNA was co-localized in immunopositive cells. They were sparsely granulated cells containing poorly developed organelles that did not resemble those of typical sparsely granulated GH cells. The findings indicate that adenoma cells are largely immature with minimal GH lineage differentiation.

Acromegaly↗

PubMed1999

Combined clear and granular cell leiomyoma of soft tissue: evidence of transformation to a histiocytic phenotype.

AIMS: We present an unusual case of leiomyoma with a clear and granular cell pattern in which there was immunohistochemical evidence of transformation to a histiocytic phenotype. METHODS AND RESULTS: A 64-year-old man presented with mild scrotal swelling and pain. A local excision was performed after the clinical diagnosis of epidermal inclusion cyst. In the pathological specimen, another tumour nodule was identified which was composed predominantly of clear cells, with an occasional mixture of granular cells. Immunohistochemical analysis demonstrated positive staining for vimentin, lysozyme, CD68 and HAM56, but complete negativity for desmin, alpha-smooth muscle actin, HHF35, S100 protein, neurone-specific enolase and CD34. Ultrastructural study revealed dilated rough endoplasmic reticulum, glycogen granules, abundant vacuolar structures and also thin microfilaments with subplasmalemmal dense bodies. CONCLUSIONS: Based on these findings, we have interpreted it to be a rare case of leiomyoma with extensive clear cell and granular cell degeneration (combined clear and granular cell leiomyoma). This complete transformation of the immunohistochemical profile into the histiocytic phenotype has not been previously described in the literature.

Actins↗

PubMed1999

Cystic lesion at the foramen magnum disseminated from a pituitary adenoma--case report.

A 38-year-old male presented with a cystic lesion at the foramen magnum due to intracranial dissemination from a pituitary adenoma. The primary tumor had required reoperation for regrowth twice. The tumor at the foramen magnum was removed surgically. Two smaller solid tumors were located in the left parietal convexity and the right temporal lobe. The former tumor was also removed surgically and the latter was observed. Histological examination showed the typical characteristics of pituitary adenoma in both surgical specimens. Immunohistochemical staining with MIB-1 and p53 antibodies showed low (< 1%) and negative reaction. Patients with pituitary adenoma, even benign tumors, must be carefully followed up for signs of metastasis.

Adenoma↗