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Biomedical subjects

M Kamei

Publications and source records attributed to M Kamei.

At least 55 records · Page 3Linked to original sources
PubMed1997

Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4.

The human monoclonal antibody (MAb) AE6F4 is secreted by a human-human hybridoma line established from the in vitro immunization of normal human peripheral blood lymphocytes with the human lung adenocarcinoma cell line, A549. This MAb is strongly reactive to lung cancer tissues. In the previous study, the antigens recognized by the MAb AE6F4 were purified from A549 cells and identified as 14-3-3 protein and 31 kDa cytosolic phospholipase A2 (cPLA2). The MAb AE6F4 also binds two kinds of antigens (53 kDa and 40 kDa), which are not related to 14-3-3 protein or 31 kDa cPLA2, in the human breast adenocarcinoma cell line, MCF-7. We purified a 38 kDa antigen, which is a degradation product of 53 kDa antigen from breast adenocarcinoma MCF-7 cells using ion-exchange and hydroxyapatite column chromatography. Two partial amino acid sequences of the purified 38 kDa antigen showed 95-100% homology to human cytokeratin 8 (CK8). Two-dimensional gel electrophoresis and immunoblot analysis of intermediate filament fraction separated from MCF-7 cells demonstrated that the 53 kDa and 40 kDa antigens were CK8 and CK19, respectively. Antigenic determinants on CK8 and CK19 recognized by the MAb AE6F4 were resistant to sodium periodate treatment, although antigenic determinant on 31 kDa antigen (14-3-3 protein and(or) cPLA2) was sensitive to this treatment. These results suggest that the MAb AE6F4 reacts with both carbohydrate and peptide antigenic determinants.

Adenocarcinoma↗
PubMed1996

POU transcription factors differentially regulate the D1A dopamine receptor gene in cultured cells.

Transcription is regulated by complex interactions between cis-elements and transcription factors. In this study, we found that members of the POU transcription factor family differentially regulate a cis-element of the human D1A dopamine receptor gene. The differences in transactivating abilities among POU factors are derived neither from their different binding affinities to the responsive element nor from their interaction with other types of transcription factors, suggesting the participation of coactivators. We also observed that POU family members, when expressed simultaneously, act competitively on this regulatory element. Our findings demonstrate that members of the same transcription factor family result in different outcomes in cultured cells. These observations suggest a novel model for in vivo gene regulation by different members of a transcription factor family through a single cis-element.

Base Sequence↗
PubMed1996

Highly hydrogenated dietary soybean oil modifies the responses to polychlorinated biphenyls in rats.

The effects of dietary highly hydrogenated soybean oil (HSO) upon the changes caused by dietary polychlorinated biphenyls (PCBs) were examined in rats. Six groups of rats were fed the following diets for 30 d: a 20% soybean oil-containing diet (control diet), a diet in which a half of soybean oil was substituted with HSO (HSO-A diet), a diet in which cellulose powder was replaced with HSO (HSO-B diet) and these diets supplemented with 100 ppm PCBs (control + PCBs, HSO-A + PCBs and HSO-B + PCBs diets). Hepatic concentration of PCBs and relative liver weight were markedly decreased in rats fed with the HSO-A + PCBs diet compared with those fed with the other diets containing PCBs. Liver lipids and liver cholesterol were considerably decreased with a reciprocal increase in fecal sterol excretion by rats fed the HSO-A + PCBs and the HSO-B + PCBs diets compared with those fed with the control + PCBs diet. The fatty acid composition in hepatic phospholipids showed an independent increase of the saturated fatty acid content induced by dietary HSO and PCBs. Dietary PCBs also caused decreases in the amounts of monounsaturated and n-3 polyunsaturated fatty acids. These results suggest that dietary HSO prevents accumulation of PCBs in the liver and promotes the excretion of lipids stimulated by PCBs, accompanied by a change in fatty acid metabolism.

Animals↗
PubMed1996

Some unusual responses of astrocytes to ghost tangles in a long duration case of juvenile Alzheimer's disease: an electron microscopic study.

The ghost tangles (GTs) in usual Alzheimer's disease are separated into small bundles of abnormal straight filaments by many invading astrocytic processes lacking a basal lamina (BL). An electron microscopic study of GTs in the Ammon's horn of a case of juvenile Alzheimer's disease of 25 years' duration, however, revealed that only a small number of astrocytic processes had infiltrated the GTs, resulting in the GTs being composed of large bundles. Moreover, the majority of glial processes that had invaded or apposed GTs possessed an interrupted but still fairly well-developed BL with hemidesmosome-like profiles. Although pia-arachnoid cells are required for astrocytes to form a continuous BL, astrocytes can have a segmental BL on their surface facing even empty intercellular spaces within the brain parenchyma. The much greater frequency and better development of the GT-associated BL in our case indicate that the GT filaments somehow increased the ability of astrocytes to form a BL. On the other hand, the scarcity of GT-invading astrocytic processes implies that many of the glial processes that had once penetrated GTs had been withdrawn, with the result that GTs escaped expectable endocyto-phagocytosis by astrocytes.

Alzheimer Disease↗
PubMed1996

Surgical removal of submacular hemorrhage using tissue plasminogen activator and perfluorocarbon liquid.

PURPOSE: To assess the result of surgical removal of submacular hemorrhage by using tissue plasminogen activator and perfluorocarbon liquid. METHODS: In 22 consecutive patients (22 eyes), subretinal hemorrhage associated with age-related macular degeneration, which involved the fovea and completely obscured the choroidal vascular pattern, was treated by pars plana vitrectomy. The hemorrhages were liquefied with tissue plasminogen activator, squeezed into the vitreous cavity with perfluorocarbon liquid, and then evacuated. RESULTS: Efficacy of the procedure was judged by the best postoperative corrected visual acuity, which was 20/100 or better in 16 eyes (73%). Submacular hemorrhage recurred in four (18%) eyes, epiretinal membrane formed in three (14%) eyes, and retinal detachment occurred in three (14%) eyes. Best-corrected final visual acuity was improved postoperatively in 18 (82%) of the 22 eyes, unchanged in three (14%) eyes, and decreased in one (5%) eye, final visual acuity was 20/200 or better in 15 eyes (68%) and limited in other eyes by subretinal hemorrhage of greater than 30 days' duration or subfoveal neovascularizations. CONCLUSIONS: Use of tissue plasminogen activator and perfluorocarbon liquid in surgical removal of submacular hemorrhage may improve the outcome of surgery by reducing surgically induced retinal damage.

Aged↗
PubMed1996

Bcl-2 inhibits retinoic acid-induced apoptosis during the neural differentiation of embryonal stem cells.

We report here that all trans-retinoic acid (RA), a classical morphogen, induces apoptosis during the neural differentiation of the embryonic stem cell line P19. The apoptotic cells showed, in addition to DNA cleavage, typical morphological changes including chromatin condensation, nuclear fragmentation, and cytoplasmic vacuolation. These apoptotic changes became obvious by 12 h after the addition of RA. The endogenous expression of bcl-2 in surviving cells was down-regulated during this process, and the compelled expression of bcl-2 by retroviral vectors reduced the number of apoptotic cells. Apoptosis was partially inhibited by adding antisense oligonucleotides against RA receptors (RARs) simultaneously or by transfecting a plasmid vector flanked with a RA-responsive element. Antisense oligonucleotides against retinoid X receptors (RXRs), the receptors for 9 cis-RA, did not inhibit apoptosis induced by all trans-RA. Cycloheximide and actinomycin D, inhibitors of protein and RNA syntheses, respectively, suppressed apoptosis. No changes were seen in the expression of tumor necrosis factors, their receptors, Fas, FasL, p53, or c-myc, molecules which have been suggested to participate in the apoptotic process. Addition of neurotrophins to the culture medium did not affect apoptosis. These findings suggest that the signals themselves, promote expression of molecules essential for apoptosis. Furthermore, we observed that RA induced apoptosis of cerebral neurons from murine embryos in primary culture, which suggests that RA might participate in cell death which occurs during neural development.

Animals↗
PubMed1996

Multiple alleles encoding a virus-like particle protein in the ichneumonid endoparasitoid Venturia canescens.

Hymenopteran endoparasitoids produce nuclear secretions from ovarian glands, which are deposited into the host insect together with the egg, protecting the developing parasitoid against the host's defence reactions. In the ichneumonid Venturia canescens, virus-like particles (VLPs), are attached to the egg surface and provide passive protection against encapsulation by the host. One of the four major particle proteins (p40) is expressed not only in the calyx gland but also in tissues that are not involved in particle production. The p40 coding DNA from V. canescens was cloned and sequenced. Within the coding DNA a tandem repeat sequence, coding for a putative proteolytic cleavage site of the PEST type, is rearranged in a significant portion of the wasp population. A corresponding polymorphism was also detected in the protein. The amino-terminal region of the deduced protein contains a putative type II transmembrane domain. The carboxy-terminal region shows similarity to the phospholipid hydroxyperoxide glutathione peroxidase (PHGPX) of vertebrates. A peroxidase function of the p40, although not ruled out, is unlikely due to the absence of a reactive centre which is typical for many vertebrate peroxidases. The overall conservation of the hydropathic region is discussed in the context of the formation of the viral envelope and its possible function in the immune protection.

Alleles↗
PubMed1996

Cytotoxicity of 7-ketocholesterol toward cultured rat hepatocytes and the effect of vitamin E.

The effects of 7-ketocholesterol on rat hepatocytes prepared by collagenase perfusion were examined. The viability of cells incubated with 100 microM 7-ketocholesterol was significantly lower than those with cholesterol, although the LDH activity in the cultured medium remained unchanged during the incubation. Hepatocytes treated with 7-ketocholesterol produced large amounts of .NO and O2- in the early stage of incubation. Treatment of the hepatocytes with Carboxy-PTIO, which selectively scavenged .NO, or with L-NMMA, an inhibitor of .NO synthase, increased the cell viability. The addition of 7-ketocholesterol to the culture medium tended to increase the ratio of total sterol to phospholipid of the hepatocytes in a time-dependent manner without changing the content of phospholipid. No lipid peroxidation or oxidation of the cellular SH groups, protein SH and glutathione, was apparent. Vitamin E added 1 h before the addition of 7-ketocholesterol prevented the hepatocytes from cell death by suppressing the incorporation of 7-ketocholesterol into the hepatocytes and by scavenging O2-.

Animals↗
PubMed1996

Production of recombinant human monoclonal antibody using ras-amplified BHK-21 cells in a protein-free medium.

A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non-protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues.

Animals↗
PubMed1996

A new wound healing model of retinal pigment epithelial cells in sheet culture.

PURPOSE: To evaluate some RPE cell functions, such as wound healing, in a preparation more similar to in situ conditions, we developed a method to obtain and culture retinal pigment epithelial (RPE) cells as a sheet. And we assessed the effects of fetal bovine serum (FBS) on the rate of RPE wound healing. METHODS: We prepared RPE sheet cultures by incubating rat eyes in 0.1% proteinase K for 13 min, peeling away the neural retina-RPE complex, and then incubating the tissue for 1 h to promote spontaneous separation of the RPE sheet from the retina. After several days of incubation, the cultured sheets of RPE cells were examined by phase-contrast microscopy, scanning and transmission electron microscopy and immunocytochemistry. We made round defects 1 mm in diameter in cultured RPE sheets and estimated the rate of wound closure in media with different concentrations of FBS (0 to 10%). RESULTS: The RPE cells cultured in sheets retained their in situ features, including microvilli, tight junctions and gap junctions, and the distribution of actin and cytokeratin filaments. A wound was noted to close with restoration of a polygonal configuration. The rate of wound closure depended on serum concentration in the culture medium; when supplemented with 10% fetal bovine serum, wound closure was complete in approximately 40 h. CONCLUSIONS: The RPE sheet-culture technique we developed thus provides a suitable model for studying such RPE cell functions as wound healing or phagocytosis.

Animals↗
PubMed1995

The rck/p54 candidate proto-oncogene product is a 54-kilodalton D-E-A-D box protein differentially expressed in human and mouse tissues.

Expression of the RCK gene, which is a target gene on 11q23 of the t(11;14) (q23;q32) translocation in the B-cell lymphoma cell line RC-K8, was studied by Northern and Western blot analyses. The RCK gene product is a member of the D-E-A-D box protein/RNA helicase family. With the use of Northern blot analysis, a 7.5-kb transcript of the RCK gene was shown to be expressed ubiquitously in human and mouse tissues. Polyclonal antibodies against the RCK gene product were raised, and the RCK gene expression pattern was examined in human and mouse tissues. Two different polyclonal anti-rck antibodies detected a specific 54-kilodalton product named rck/p54 in the majority of human and mouse tissues tested by Western blot analysis. However, rck/p54 was shown to be very low in the human brain and was not detectable in lumbar muscle and lung tissues, although RCK mRNA is abundantly present in these tissues. It is of interest that malignant transformed human cells arising from tissues with low or no expression of rck/p54, such as neuroblastoma, glioblastoma, rhabdomyosarcoma, and lung cancer cell lines, produced a moderate amount of rck/p54 protein, suggesting that rck/p54 plays a role in tumorigenesis. In addition, the rck/p54 protein was localized to cytoplasm by immunostaining with the use of laser microscopy and by subcellular fractionation.

Amino Acid Sequence↗
PubMed1995

Effects of highly hydrogenated soybean oil and cholesterol on plasma, liver cholesterol, and fecal steroids in rats.

We investigated the relationship between dietary highly hydrogenated soybean oil (HSO) and cholesterol transport in rats. In the first study, to examine the effects on cholesterol transport of different concentrations of HSO in dietary oil, rats were given one of the three diets containing 0, 25, or 50% HSO in dietary oil with cholesterol (5 g/kg diet) or a diet without HSO and cholesterol for 22 d. Feeding the high concentration of HSO prevented the increase in plasma total cholesterol, hepatic total lipids, and cholesterol and the decrease in high-density lipoprotein-cholesterol, which were caused by dietary cholesterol. Moreover, HSO increased the fecal excretion, fecal lipids, and steroids in a dose-dependent manner. In the second study, to examine the effects on cholesterol transport of redistribution of steric acid in the triacylglycerol species contained in HSO, rats were given one of the six diets containing HSO (distearoylmonoacylglycerol and tristearoylglycerol)-rich, monostearoylglycerol-rich, or palmitic acid-rich oil with/without cholesterol (5 g/kg diet), for 30 d. Whereas the accumulation of cholesterol in the body was reduced, cholesterol excretion was enhanced effectively in rats given the HSO-rich diet compared with rats given the monostearoylglycerol-rich diet. These results suggested that not only the high concentration of stearic acid but also its uneven distribution in HSO-triacylglycerol contributed to the reduction in intestinal cholesterol absorption in rats.

Animals↗
PubMed1995

Rupture of congenital peripheral pulmonary aneurysm.

A 58-year-old man presenting with solitary aneurysm of a peripheral pulmonary artery was treated by left lower lobectomy. Histologically, the aneurysmal wall showed medial hypertrophy with the loss of smooth muscle fibers, without evidence of a mycotic process or inflammatory exudate. The aneurysm appeared to be congenital in origin.

Aneurysm↗
PubMed1995

A rapid method for isolation of retinal pigment epithelial cells from rat eyeballs.

A new, simple method for isolating retinal pigment epithelial (RPE) cells from rat eyeballs was developed: incubation of the eyeballs in 0.1% proteinase K solution. This method yielded an average of 4 x 10(4) viable RPE cells from each Sprague-Dawley rat eyeball in 1 h. These RPE cells were hexagonal and had cytokeratin in their cytoplasm and numerous microvilli on their surface, similar to RPE cells evaluated in vivo. The results of this study thus show that our method for incubating eyeballs in proteinase K can provide good-quality RPE cells in sufficient quantities for study in a short time.

Animals↗
PubMed1995

Congenital renal glomerular fibrosis in a case of swine chondrodysplasia.

A dwarf male Large White neonate showing various external abnormalities was diagnosed to be chondrodysplasia. Microscopy revealed massive accumulation of eosinophilic homogeneous substance in the renal glomeruli, mainly along the capillary walls. The homogeneous substance, mainly consisted of type-III collagen, was not found in the primitive glomeruli in the superficial cortex, and was most frequently and intensely deposited in the mature glomeruli in the deep cortex. Electron microscopically, the deposits consisted of the collagen fibrils with various structural abnormalities. Glomerular fibrosis might be attributed to in situ production of type-III collagen by glomerular cells with differentiation and maturation, most likely by the mesangial cells.

Animals↗
PubMed1995

Immunocytochemical detection of lung cancer cells with monoclonal antibodies to 14-3-3 proteins.

Murine monoclonal antibodies were raised against 14-3-3 proteins, the antigen of human monoclonal antibody AE6F4 which had been shown potentially useful for the immunochemical diagnosis of lung cancer via sputum cytology. Enzyme-linked immunosorbent assays of the murine anti-14-3-3 monoclonal antibodies with isolated bovine brain 14-3-3 isoforms showed that the antibodies were classified into four different profiles of isoform reactivity. The comparison of 14-3-3 isoform and lung cancer tissue on the reactivity with murine monoclonal antibodies indicated that beta isoform can be responsible for cancer recognition, whereas human monoclonal antibody AE6F4 showed preferential binding to zeta isoform. No murine monoclonal antibody of the same isoform specificity as human monoclonal antibody AE6F4 was obtained. Since murine monoclonal antibodies with different isoform specificities could immunostain lung cancer cells in sputum successfully, the combination use of murine monoclonal anti-14-3-3 antibodies with human monoclonal antibody AE6F4 is potentially useful for facilitating the sputum cytodiagnosis of lung cancer.

14-3-3 Proteins↗