Isamu Kino

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 °C, 2 atm for 20 min), microwave oven (in water), and simple heating (60 °C overnight in distilled water or 90 °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.

CYP1A1 and CYP2E1 polymorphism and lung cancer, case-control study in Rio de Janeiro, Brazil

Msp I polymorphism and exon 7 Ile-Val polymorphism of CYP1A1, and Rsa I polymorphism of CYP2E1 were studied in lung cancer patients and controls in Rio de Janeiro, Brazil. Of the three polymorphisms studied, only the exon 7 polymorphism of CYP1A1 (Val-containing genotypes) had a distribution which was statistically significant in the patients and controls. The contribution of Val containing genotypes of CYP1A1 exon 7 was greater in the subpopulation of squamous cell carcinoma patients with a lower life-time smoking consumption (OR, 2.92 vs 1.97). This association is consistent with the previous findings by Kawajiri et al. and the first observation of the positive association of this locus with lung cancer in a Western population (Kawajiri K, Nakachi K, Imai K, Yoshii A, Shimada N, Watanabe J. FEBS Let 1990; 263, 131-133). Furthermore, together with the lack of association of Msp I polymorphism in the non-coding region of CYP1A1, the locus truly responsible for lung cancer risk among pleural polymorphisms of CYP1A1 appeared to be exon 7 Ile-Val polymorphism. In the future, investigations of multiple markers in different ethnic populations may reveal cancer risk markers common to all mankind.

Inflammatory myoglandular polyps of the colon and rectum, A clinicopathological study of 32 pedunculated polyps, distinct from other types of polyps

Hitherto unclassified colorectal polyps were identified in 32 patients (23 men and 9 women; mean age, 53 years). The only symptom, which was observed in less than half the patients, was passage of blood or occult blood. Endoscopic examination revealed solitary pedunculated, red polyps with a smooth surface. These polyps were found in the left colon, especially in the sigmoid. Their characteristic features were inflammatory granulation tissue in the lamina propria mucosae, proliferation of smooth muscle, and hyperplastic glands with occasional cystic dilatation. The etiology of this type of polyp is unknown, but it could involve chronic trauma from the fecal stream and from peristalsis of the bowel. These polyps can be differentiated from juvenile polyps and inflammatory polyps by the presence of abundant smooth-muscle cells in the inflamed lamina propria mucosae. They also can be differentiated from Peutz-Jeghers polyps, which appear as hamartomatous structures with tree-like proliferation of muscularis mucosae covered by colonic mucosa without inflammatory granulation tissue. Their locations and macroscopic appearance distinguish these polyps from mucosal prolapse syndrome and polyps developed after colostomy. In addition, these new polyps differ from inflammatory cap polyps in that they lack a fibrin cap. We propose the name inflammatory myoglandular polyps for these polyps, which are distinct clinicopathologically from other types of colorectal polyps.

Localization of Menkes gene expression in the mouse brain; its association with neurological manifestations in Menkes model mice

Abstract Menkes gene (Mc1 or MNK, encoding putative copper-transporting ATPase) expression was investigated and compared in normal and macular mutant mouse brain. Northern blot analysis showed a distinct 8.3-kb transcript and no obvious difference in size or extent in normal mice and macular mutants on postnatal days 0, 4, 7, 10 or 13. In situ hybridization revealed that certain specific populations of cells in the brain express Menkes mRNA, and that their localization in normal and mutant mice did not differ and was conserved on days 4, 10 and 13. The most intense hybridization signals were observed in the hippocampal CA1 region and dentate gyrus, the olfactory bulb nuclei, the cerebellar granular cell layer, the choroid plexus and the ependyma, with less intense signals in the hippocampal CA3 region and cerebellar Purkinje cells. In addition, necrotic neuronal cell death was predominantly observed in the CA3 region and the Purkinje cells of macular mice after postnatal day 10. The finding that the regions that had lower expression level of Menkes mRNA corresponded to those showing neuronal necrosis suggests that the Menkes gene may be responsible for the neuronal degeneration in some specific portions of the brain and clinical manifestations in this mutant.

Morphometrical and Cell Kinetic Studies of Normal Human Colorectal Mucosa

Morphometric and cell kinetic studies were performed on normal mucosa in 30 specimens of surgically resected colorectum. The average crypt length was 372 ± 57μm in the proximal colon and 506 ± 65μm in the distal colon, and the average cell numbers per crypt column in these regions were 76.4 ± 8.9 and 80.3 ± 8.8, respectively. In the proximal and distal colon, goblet cells constituted 23.3 ± 3.4% and 25.2 ± 4.0%, respectively, of the total cells in each crypt, and in both regions entero chromaffin cells constituted about 1%. The labeling indices of the proximal and distal colon were 9.4 ± 3.9% and 8.2 ± 2.2%, respectively. Combined microautoradiography and periodic acid Schiff reaction on selected cases revealed that the labeling indices of columnar cells and goblet cells were 11.0 ± 2.0% and 7.0 ± 3.2%, respectively. It was suggested that the increased number of larger goblet cells in crypts of the sigmoid colon and rectum resulted in an increase of crypt length and a decrease in the labeling index. These results provide information on the morphological and cell kinetic characteristics of normal mucosa of the colon and rectum. Acta Pathol Jpn 39: 725 730, 1989.

Promotion of collagen production by human fibroblasts with gastric cancer cells in vitro

SummaryTo elucidate the histogenesis of gastric scirrhous cancer, the promotion of collagen production by normal human skin fibroblasts (HSF-1) with human gastric cancer cells (KATO-III, MKN-45 and MKN-28) was investigated by direct coculture and parabiotic culture.Argyrophilic collagenous fibers were demonstrated among fibroblasts on both direct cocultures and parabiotic cultures of the fibroblasts with gastric cancer cells. Microscopic examination showed that these fibers appeared earlier and were more abundant and thicker in direct cocultures and parabiotic cultures than in single cultures of fibroblasts. Gastric cancer cells in single or parabiotic culture did not form argyrophilic fibers.For quantitative proof of the promotion of collagen production by fibroblasts with gastric cancer cells, hydroxyproline produced by fibroblasts was measured. Much higher fibroblast hydroxyproline values were obtained in parabiotic cultures with gastric cancer cell lines than in single cultures of HSF-1. Moreover, the rate of collagen synthesis by HSF-1 was much higher than that of any gastric cancer cell line tested.These results demonstrate that gastric cancer cells enhance collagen production by fibroblasts in vitro. This finding suggests that they may produce a factor promoting fibroblast collagen synthesis and that this may contribute to the formation of stromal collagen in human gastric scirrhous cancer.

Minute adenomas of the depressed type in familial adenomatous polyposis of the colon. A pathway to ordinary polypoid adenomas.

Background. It has been thought that colorectal adenomas are polypoid from their beginning. Although depressed adenomas have been described, they have been considered an unusual form and have not been studied systemically.

Effects of pyrroloquinoline quinone (PQQ) and PQQ-oxazole on DNA synthesis of cultured human fibroblasts

The effects of pyrroloquinoline quinone (PQQ) and PQQ-oxazole (PQQ-glycine adduct) on DNA synthesis were examined using cultured human fibroblasts. Confluent fibroblasts were cultured in serum-free Dulbeccos modified Eagles media, and various concentrations of PQQ and PQQ-oxazole were added to the media. After incubation for 24 h, [3H]thymidine was added to the media as an indicator for DNA synthesis of the cells. The thymidine incorporation into the cells was significantly enhanced even in the presence of very low concentrations of PQQ (0.003-0.03 microM); it remained significantly enhanced, up to 30 microM PQQ. However, the incorporation remarkably decreased at 750-1500 microM of PQQ. In contrast to the results for PQQ, DNA synthesis was not stimulated by addition of 0.003-3 microM PQQ-oxazole, but it was slightly enhanced at concentrations 15-750 microM. In morphological examination of the cultured human fibroblasts, cell density was increased by addition of 0.003-30 microM PQQ when compared with that of the control, supporting the above biochemical data. However, there were no distinct differences in morphological effect between PQQ and PQQ-oxazole.

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.

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.