Biomedical subjects

I Kino

Publications and source records attributed to I Kino.

At least 37 records · Page 2Linked to original sources

PubMed1995

Efficient and specific induction of esophageal tumors in rats by precursors of N-nitrososarcosine ethyl ester.

Cancers and precancerous lesions of the esophagus were efficiently induced in rats by the simulation of a clinico-epidemiological setting; that is, the administration of precursors of nitrosamine. Six week old non-inbred male Wistar rats were given 2g/kg bodyweight of sarcosine ethyl ester hydrochloride (SEEH) and concurrently 0.3g/kg bodyweight of sodium nitrite (NaNO2), precursors of N-nitrososarcosine ethyl ester (NSEE), in 2% sucrose as drinking water. Group 1 received the precursors twice a week for 6 weeks followed by 8 weeks observation, and group 2, once every 3 days for 7 weeks followed by 26 weeks observation. At the end of treatment, no tumor had developed in the esophagus of rats in group 1, but the [3H]-thymidine labeling indices in both basal and superficial layer cells were higher than in the control group. On subsequent observation, papillomas appeared in group 1 (33.3%), and carcinomas in group 2 (33.3%), within 4 weeks. The tumors induced in group 1 were mostly papillomas and rarely carcinomas. When the observation was prolonged in group 2, 100% of the animals had cancer in week 20. The pathological changes of the lesions paralleled the sequential development of human squamous cell carcinoma of the esophagus. Our system has the advantages in that papillomas and cancers can be induced in rats in a short time and the agents used are less toxic than preformed nitrosamines administered previously by gastric intubation. It would serve as a useful experimental tool to study premalignant lesions and cancers of the esophagus.

Animals↗

PubMed1995

Monoclonality of normal human colonic crypts.

The monoclonality of human colonic crypts was demonstrated by human androgen receptor (HUMARA) gene assay following application of the crypt isolation method. DNA was extracted from an isolated single crypt, Hpa II digestion was performed before polymerase chain reaction (PCR) by primers spanning the HUMARA exon 1 region. The PCR product of a single crypt clearly showed allelic exclusion based on methylation status, while PCR product from a mixture of 40 crypts or colonic mucosa as a whole that included epitheliums and interstitial connective tissue had two bands. This method will facilitate the non-isotopic analysis not only of tumor clonality, but also of the normal structures derived from a single progenitor cell.

Base Sequence↗

PubMed1995

Cytophotometric and flow cytometric DNA content of isolated glands in gastric neoplasia.

The gland isolation method was applied to various gastric lesions to measure DNA content by cytophotometry and flow cytometry for the first time. By incubating and agitating fresh specimens from surgically resected stomachs in calcium-magnesium free Hanks's balanced salt solution (CMFH) containing EDTA, many neoplastic glandular epithelial cells were successfully isolated from the stroma, and their characteristic three dimensional features were seen morphologically. The DNA content of pure nuclear suspensions of isolated glands was obtained by cytophotometry and flow cytometry staining with 4',6-diamino-2-phenylindole dihydrochloride (DAPI) and propidium iodide, respectively. Compared with histological grading, the frequency of the DNA aneuploidy of cancer with moderate or poor differentiation by cytophotometry (75%) was significantly higher than that of well differentiated cancer (25%), but the histological typing of gastric cancer DNA frequency were not correlated. This method allowed us to detect small aneuploid peaks by flow cytometry, which were previously masked by contaminating interstitial cells. The frequency of DNA aneuploidy detected by flow cytometry (87.5%) was higher than detected by cytophotometry (58.3%). The results of these studies shows the feasibility of this technique for analysing the DNA content of various lesions of the stomach.

Adenocarcinoma↗

PubMed1995

Immunocytochemical study of microvilli in a gastric carcinoma-derived cell line.

To study the structural components of microvilli of the KATO-III cell, we used anti-villin, -ezrin, and anti-MVM (microvillous membrane prepared against mouse intestinal microvilli) antibodies. Villin and ezrin cross-link actin bundles of microvilli such as those in the small intestine and renal proximal cells. Electromicroscopically, the cytoskeletal core of microvilli of the KATO-III cell was constituted of actin-filament bundles. The anti-villin antibody but not anti-ezrin antibody reacted with the KATO-III cell as demonstrated by FITC-immunofluorescence and PAP-staining. Anti-villin, anti-MVM, but not anti-ezrin antibody, reacted with the KATO-III cell surface and with intracellular materials. Western blot analysis using anti-villin and anti-MVM antibodies revealed proteins of approximately 95 kDa (villin), and 15 kDa in the microsomal membrane fractions of KATO-III cells, respectively. Immunocytochemical and confocal laser microscopic studies showed that the cell-surface and the intracellular microcysts of KATO-III cells were preferentially decorated by anti-villin and anti-MVM antibodies. These data suggested that some actin-binding proteins, such as villin, were localized at the cell surface and on some of the intracellular cytoplasmic structures of the KATO-III cell.

Animals↗

PubMed1995

[Immunohistochemical distribution of p53 and Ki-67 in 2 cases of colorectal carcinoma occurring in ulcerative colitis].

We studied immunohistochemically the distribution of p53 overexpression and Ki-67 as a marker of cell proliferation, in 2 cases of advanced colorectal carcinomas occurring in UC, in comparison with histopathological features such as carcinoma and dysplasia. One is a case of 4 independent colorectal carcinomas surrounded by widespread dysplasia, and the other is a case of solitary rectal carcinoma. In these 2 cases, formalin-fixed paraffin-embedded tissues covering all of the resected specimens were examined for p53 and Ki-67 immunoreactivity using microwaving technique. p53 was negative in normal mucosa and inflammative mucosa without dysplasia. p53 was strongly positive in cancerous lesions except one lesion, and various degrees of dysplasia surrounding these lesions also showed p53 positive. The numbers of p53 and Ki-67 positive cells were higher in severe dysplasia than in mild dysplasia. Some areas of the dysplastic mucosa histologically represented "hyperplastic pattern of villous feature." In these areas p53 positive cells were aggregated in the basal part of the villous crypt, and the number of Ki-67 positive cells per gland was larger than in normal gland. Our examination covering the whole mucosa revealed: 1) p53 and Ki-67 are useful diagnostic markers for grading the degree of dysplasia. 2) Positivity of immunoreactive p53 in dysplasia seems to be consistent with that in cancers occurring in the surrounding dysplastic area in most of our cases.

Adenocarcinoma, Mucinous↗

PubMed1995

A case of triple early gastric cancer in the remnant stomach.

A 63-yr-old man, who had undergone distal gastrectomy and gastrojejunostomy (Billroth II method) 32 yr previously for duodenal ulcer, was admitted with suspected gastric remnant cancer. An upper gastrointestinal series and endoscopy revealed a protruding lesion at the stoma of the remnant stomach. Total gastrectomy with resection of the adjacent jejunum was performed. Histological examination demonstrated two well-differentiated adenocarcinomas (a mixed type I and IIc lesion with submucosal invasion and a type IIa lesion with intramucosal invasion) and a type IIc mucocellular carcinoma located in the mucosa. Gastritis cystica polyposa also was observed in the remnant stomach. The combination of three early gastric cancers and gastritis cystica polyposa suggests that the mucosa of the remnant stomach had a high malignant potential. The patient has survived without recurrence for 5 yr since the operation.

Adenocarcinoma↗

PubMed1995

Histological classification of the neoplastic changes arising in ulcerative colitis: a new proposal in Japan.

Patients with total ulcerative colitis with a longstanding course of the disease have a high risk of developing colorectal carcinoma. Colonoscopic surveillance to detect precancerous tissue and/or cancer in these patients has been carried out in countries with a high incidence of ulcerative colitis. Riddell's classification has been widely used for the interpretation of biopsy specimens obtained from the colonoscopic surveillance. In Japan, however, there are problems in accepting Riddell's classification, mainly because the intramucosal carcinomas diagnosed by Japanese histopathologists are included in the category of high-grade dysplasia in Riddell's classification. Based on the results of a meticulous slide review carried out by seven histopathologists in this study, a new classification is proposed: UC-I, inflammatory change; UC-II, indefinite; UC-IIa, probably inflammatory; UC-IIb, probably neoplastic; UC-III, neoplastic but not carcinomatous; and UC-IV, carcinoma. Intramucosal carcinomas is included in the category UC-IV. We consider that the diagnosis of intramucosal carcinoma is to be made when there is a high grade of cytological and structural atypia consistent with carcinoma. Interobserver and intraobserver variability with this classification was acceptable. We believe this new classification will be widely use in cancer surveillance in ulcerative colitis in Japan.

Biopsy↗

PubMed1994

High prevalence of p53 protein overexpression in patients with esophageal cancer in Linxian, China and its relationship to progression and prognosis.

BACKGROUND: Linxian is the highest endemic area of esophageal squamous cell carcinoma (ESCC) in China and one of the highest incidence areas in the world. The relationship of p53 protein accumulation to geographic variation, pathologic findings, and prognosis has not been investigated extensively. METHODS: Formalin fixed, paraffin embedded ESCC tissues from 100 patients who underwent esophagectomy between 1973 and 1983 were immunostained by using monoclonal antibody pAB1801. RESULTS: p53 overexpression was observed in 41 (87.2%) of 47 tumors of patients in Linxian and in 16 (64%) of 25 additional patients outside Linxian. Its prevalence in the noncancerous epithelium (11/72, 15.3%) and carcinoma in situ (1/7, 14.3%) was lower than that in invasive lesions (64/93, 68.8%). Its immunostaining intensity increased with the depth of cancer invasion. Of 30 primary carcinomas with lymph node metastasis, 29 (96.7%) were positive. However, only 36 (51.4%) of 70 primary lesions without metastasis were positive, and a higher intensity was noticed in the metastases. There was a lower expression rate in tumors of patients surviving more than 10 years (25/52, 48.1%) than in those surviving less than 3 years (40/48, 83.3%). Overall and nonadvanced or metastasis-free cumulative survival rates were both significantly different in patients with and without p53 protein overexpression. CONCLUSIONS: There is a higher expression rate of p53 protein in ESCC in tumors of patients from Linxian than in those from the surrounding area. The accumulation of p53 protein is related to the invasiveness and capability for metastases of cancer cells and appears to be a useful prognostic factor for patients with ESCC.

Adult↗

PubMed1994

Overexpression of ERK, an EPH family receptor protein tyrosine kinase, in various human tumors.

The ERK gene has been isolated as a genomic DNA encoding a part of the receptor protein-tyrosine kinase which belongs to the EPH subfamily. We previously identified a partial complementary DNA (cDNA) encompassing the catalytic domain of ERK from the expression library of human gastric cancer with an antiphosphotyrosine antibody. Using this cDNA as a probe, the cDNAs encoding mature ERK protein were isolated. The putative mature ERK protein, a total of 967 deduced amino acid residues, showed high homology with chicken Cek5 (92.5%) and mouse Nuk (99.1%). Chromosomal in situ hybridization revealed that human ERK cDNA is localized to chromosome 1p34-35. In Northern blot analysis of normal human tissues, the ERK gene was ubiquitously expressed mainly in cells of epithelial origin but not in the brain. Studies on RNAs from 76 human tumor tissues and cell lines showed that ERK is expressed at higher levels in various tumors of epithelial origin than in corresponding normal tissues, most frequently in gastric cancers (12 of 16, 75.0%). Overexpression of ERK was also detected in one osteosarcoma cell line. These findings suggest that ERK plays some significant role in carcinogenesis in the stomach and other tissues.

Amino Acid Sequence↗

PubMed1994

Isolation of complementary DNA encoding K-cadherin, a novel rat cadherin preferentially expressed in fetal kidney and kidney carcinoma.

Complementary DNA for a novel member of the cadherin family, designated K-cadherin, was isolated from a rat renal cell carcinoma complementary DNA library by screening it with a short complementary DNA probe which was initially obtained from the RNA of day 16 fetal Wistar rat stomach mucosa by the polymerase chain reaction. The deduced primary structure of K-cadherin is 789 amino acid residues, which contain five internal repeats in its extracellular domain, a single putative transmembrane domain, and a cytoplasmic tail characteristic of those of classic type cadherins. K-cadherin exhibits low homology with mature proteins of mouse N- (38%), E- (35%), and P-cadherin (32%), and high homology with a partially identified human cadherin-6 protein (95%) at the amino acid level. Northern blot analysis revealed a high level of expression of K-cadherin mRNA in fetal rat kidney and brain, and rat kidney carcinoma with two major transcripts, 4.1 and 8.0 kilobases in size, whereas there was very weak or no expression in any organ of adult rats. The level of K-cadherin expression was also elevated in some human kidney cancer tissues. In the developing kidney, in situ hybridization showed localization of K-cadherin mRNA in the nephroblastic epithelial cells of comma bodies coinciding with those in the process of polarization during glomeruloneogenesis. These results demonstrate that K-cadherin must have important functions in both the process of kidney development and tumorigenesis of some types of kidney cancer.

Amino Acid Sequence↗

PubMed1994

Mapping of the human C3G gene coding a guanine nucleotide releasing protein for Ras family to 9q34.3 by fluorescence in situ hybridization.

C3G, a human guanine nucleotide releasing protein for Ras protein, was mapped to human chromosome 9q34.3 by fluorescence in situ hybridization with R-banded chromosomes. C3G was originally identified as one of the CRK-binding proteins, similar to c-abl (9q34.1). Our result suggests that the downstream factors of Crk are localized in close proximity on chromosome 9.

Chromosome Banding↗

PubMed1994

Metastases from gastric adenocarcinoma presenting as multiple colonic polyps: report of a case.

A 53-year-old woman presented with symptoms of weight loss, diarrhea, and melena. A barium enema with endoscopy revealed multiple colonic polyps which were shown histologically to be metastatic deposits of poorly differentiated adenocarcinoma. The primary tumor, a poorly differentiated gastric adenocarcinoma, had been resected 11 years earlier. This appears to be only the second published report of polypoid colonic metastases from gastric adenocarcinoma.

Adenocarcinoma↗

PubMed1994

Detection of minute intestinal metaplastic lesions by video microscopy.

Using video microscopy in 225 resected stomachs we detected 32 minute solitary lesions of intestinal metaplasia histologically. The magnified features of the minute lesions showed a characteristic appearance and were classified into three types: mesh-like (type A), villoid (type B), and tubular (type C). All the lesions classified as type A exhibited the incomplete type of intestinal metaplasia according to the results of histopathological and/or histochemical examination. In contrast, most lesions classified as type B exhibited the complete type of intestinal metaplasia. We concluded that intentional detection of minute lesions in resected stomachs by video microscopy is simple and useful, especially in cases of minute lesions < 5 mm in diameter. Moreover, our findings demonstrate that the minute intestinal metaplastic lesions have morphological characteristics based on which they can be classified into three types of lesions. These morphological characteristics correlate with their histopathological findings.

Adult↗

PubMed1994

Application of the crypt-isolation technique to flow-cytometric analysis of DNA content in colorectal neoplasms.

BACKGROUND/AIMS: Conventional flow-cytometric analysis of colorectal tumor DNA content includes both tumor and stromal cell DNA. To improve the method, we evaluated the DNA content of purified epithelial cells using the crypt-isolation technique. METHODS: Normal and neoplastic crypts were isolated from fresh resected colorectum nonenzymatically. Isolated crypts were digested into single nuclei suspensions and examined by flow cytometry (FCM). Internal controls from normal crypts were used to define diploidy. The neoplastic nuclei in the same individual and mixtures of both normal and neoplastic nuclei in various ratios were analyzed. RESULTS: Tumors having both diploid and aneuploid stemlines were found more frequently than that reported using conventional FCM. Near-diploid DNA stemlines with DNA indices ranging between 0.90 and 1.10 were detectable. The cases of advanced cancer in which the DNA index was between 0.90 and 0.95 were associated significantly with Dukes' stage C. CONCLUSIONS: This is the first reported analysis of the DNA content of "pure" neoplastic colorectal epithelia using FCM. Multiploid and near-diploid stemlines were more accurately analyzed than that using conventional FCM.

Adenocarcinoma↗

PubMed1994

Benign schwannoma of the esophagus: report of two cases with immunohistochemical and ultrastructural studies.

Two cases of benign schwannoma of the esophagus are presented. The tumors were found in the thoracic esophagus of women of 56 and 64 years of age, respectively, who had complained of dysphagia and back pain. Tumorectomies were performed and the tumors were found to be located within the esophageal wall arising from the muscularis propria. The tumors were examined immunohistochemically and ultrastructurally. These tumors were identical in gross, histological and electron microscopic features. Grossly, the tumors showed yellowish-white cut surfaces without hemorrhage or necrosis. Microscopically, they were composed of spindle-shaped cells showing moderate variation in size and shape, and nuclear palisading. Lymphoid aggregates with germinal centers surrounded the tumors. Immunohistochemically, strong reactions for S-100 protein and neuron-specific enolase were observed in the cytoplasm of spindle cells, whereas reactions for muscle actin and desmin were negative. These findings, together with electron microscopic observations, supported the Schwann cell origin of these tumors.

Esophageal Neoplasms↗

PubMed1994

Histochemical and ultrastructural analyses of glandular differentiation in typical carcinoid tumor of the hindgut.

Adenomatous differentiation of typical carcinoid tumors was examined by histochemical and ultrastructural studies on 33 cases of typical carcinoids of the hindgut. Of these 33 carcinoids, 12 (36%) showed a tubular and/or acinar pattern with production of mucus as a minor component. A few tumor cells with mucous globules were scattered among tubules in these carcinoids. Ultrastructurally, microvilli were seen along the luminal surface of tubules, with both numerous mucous globules in the supranuclear area and endocrine granules in basal sites of the same cells. These results indicate that even in typical carcinoid tumors, certain tumor cells have the features of goblet cells or the open type of endocrine cells. The findings of both mucous globules and endocrine granules in the same single cells indicate a close relationship between endocrine cells and mucus-producing and/or tubule-forming cells. It is concluded that pluripotential cells producing carcinoid tumors might be able to differentiate into three types of tumor cells: (i) endocrine cells without microvilli or mucus production; (ii) cells with microvilli and no mucus product; and (iii) mucus-containing cells with microvilli.

Adult↗

PubMed1994

A morphometrical analysis of minute depressed adenomas in familial polyposis coli.

In a morphometrical study of minute adenomas in familial polyposis coli (FPC), ranging from 0.8 mm to 1.5 mm in diameter, five parameters of depressed adenomas were compared with those of ordinary polypoid adenomas. The density of glands (DG), an index of structural atypia, and the nuclear-cytoplasmic (N/C) ratio, an index of cellular atypia, were higher in depressed adenomas than in polypoid adenomas (74.8% vs 66.8% and 37.8% vs 32.7%, respectively). The cell area (C-area) of depressed adenomas was less than that of polypoid adenomas (107.3 microns 2 vs 121.1 microns 2), but there was no significant difference in values for the nuclear area (N-area) in the two types (40.0 microns 2 vs 39.2 microns 2). The higher N/C ratio of depressed adenomas was, therefore, not caused by enlargement of the nuclei but by reduction of the cell size. In addition, values of the sphericity of nuclei (SN) (3.14 vs 3.65) indicated rounder nuclei in depressed adenomas than in ordinary ones. The higher DG and N/C ratio of the depressed adenomas indicates their higher grade of atypia than that of ordinary polypoid adenomas.

Adenoma↗

PubMed1994

Alteration of immunoreactivity by hydrated autoclaving, microwave treatment, and simple heating of paraffin-embedded tissue sections.

The effects of treatment in a hydrated autoclave (121 degrees C, 2 atm for 20 min), microwave oven (in water), and simple heating (60 degrees C overnight in distilled water or 90 degrees C for 10 min in ZnSO4) on the stainability of 56 antigens by commercially available antibodies in formalin-fixed paraffin-embedded tissue sections were evaluated. The detectability of nuclear antigens, glycoprotein, lymphocytic surface markers, and chromogranin A was significantly and reproducibly improved by these treatments, whereas the detectability of viral antigens and peptide hormones was attenuated or unchanged. This enhancement includes not only the distinctiveness of the positive staining, but also the number of positive cells, as revealed by comparing serial sections. Among these four heating procedures, microwave heating and autoclaving were more effective than the others on p53, c-erbB-2, and CA125, whereas simple heating was best for smooth-muscle actin (HHF35 and CGA7). Generally the effects of the heating procedures for these antigens were consistent among the cases, but the effects on GFAP varied with the case. The alterations we observed could significantly influence the interpretation of immunohistochemical staining of currently popular tumor markers such as p53 in terms of their prevalence (28% vs 64% in gastric cancer; 36% vs 82% in metastatic liver cancer) and other diagnostically important markers.

Antigen-Antibody Reactions↗