Biomedical subjects

T Kinugasa

Publications and source records attributed to T Kinugasa.

At least 19 recordsLinked to original sources

PubMed2001

Interleukin-2 receptor beta subunit-dependent and -independent regulation of intestinal epithelial tight junctions.

Interleukin (IL)-15 is able to regulate tight junction formation in intestinal epithelial cells. However, the mechanisms that regulate the intestinal barrier function in response to IL-15 and the involved subunits of the IL-15 ligand-receptor system are unknown. We determined the IL-2Rbeta subunit and IL-15-dependent regulation of tight junction-associated proteins in the human intestinal epithelial cell line T-84. The IL-2Rbeta subunit was expressed and induced signal transduction in caveolin enriched rafts in intestinal epithelial cells. IL-15-mediated tightening of intestinal epithelial monolayers correlated with the enhanced recruitment of tight junction proteins into Triton X-100-insoluble protein fractions. IL-15-mediated up-regulation of ZO-1 and ZO-2 expression was independent of the IL-2Rbeta subunit, whereas the phosphorylation of occludin and enhanced membrane association of claudin-1 and claudin-2 by IL-15 required the presence of the IL-2Rbeta subunit. Recruitment of claudins and hyperphosphorylated occludin into tight junctions resulted in a more marked induction of tight junction formation in intestinal epithelial cells than the up-regulation of ZO-1 and ZO-2 by itself. The regulation of the intestinal epithelial barrier function by IL-15 involves IL-2Rbeta-dependent and -independent signaling pathways leading to the recruitment of claudins, hyperphosphorylated occludin, ZO-1, and ZO-2 into the tight junctional protein complex.

Base Sequence↗

PubMed2001

Initial symptoms of acute radiation syndrome in the JCO criticality accident in Tokai-mura.

A criticality accident occurred on September 30, 1999, at the uranium conversion plant in Tokai-mura (Tokai-village), Ibaraki Prefecture, Japan. When the criticality occurred, three workers saw a "blue-white glow," and a radiation monitor alarm was sounded. They were severely exposed to neutron and gamma-ray irradiation, and subsequently developed acute radiation syndrome (ARS). One worker reported vomiting within minutes and loss of consciousness for 10-20 seconds. This worker also had diarrhea an hour after the exposure. The other worker started to vomit almost an hour after the exposure. The three workers, including their supervisor, who had no symptoms at the time, were brought to the National Mito Hospital by ambulance. Because of the detection of gamma-rays from their body surface by preliminary surveys and decreased numbers of lymphocytes in peripheral blood, they were transferred to the National Institute of Radiological Sciences (NIRS), which has been designated as a hospital responsible for radiation emergencies. Dose estimations for the three workers were performed by prodromal symptoms, serial changes of lymphocyte numbers, chromosomal analysis, and 24Na activity. The results obtained from these methods were fairly consistent. Most of the data, such as the dose rate of radiation, its distribution, and the quality needed to evaluate the average dose, were not available when the decision for hematopoitic stem cell transplantation had to be made. Therefore, prodromal symptoms may be important in making decisions for therapeutic strategies, such as stem-cell transplantation in heavily exposed victims.

Amylases↗

PubMed2000

Claudins regulate the intestinal barrier in response to immune mediators.

BACKGROUND & AIMS: To determine the functional role of immune mediators in the formation of the intestinal barrier, we have examined the regulation of claudin expression by interleukin (IL)-17 in human intestinal epithelial cells. METHODS: Expression of claudins, extracellular signal-related (ERK) mitogen-activated protein kinases (MAPKs), and activated ERK MAPKs was determined by immunoblotting. Claudin membrane association was assessed by immunohistochemistry and claudin messenger RNA expression by Northern blot analysis. Intestinal epithelial barrier function was characterized through transepithelial electrical resistance and mannitol tracer flux. RESULTS: IL-17 induced the development of a paracellular barrier of T84 cell monolayers. Inhibition of ERK activation with the MEK inhibitor PD98059 blocked IL-17 as well as basal development of tight junctions in T84 cells. IL-17 induced formation of tight junctions correlated with up-regulation of claudin-1 and claudin-2 gene transcription. Inhibition of MEK reduced the activated and basal expression of claudin-2 messenger RNA and protein expression. Functional MEK was required for the expression and membrane association of claudin-2 but not claudin-1 in T84 cells. CONCLUSIONS: MEK activity is required for claudin-mediated formation of tight junctions. IL-17 is able to regulate the intestinal barrier through the ERK MAPK pathway.

Blotting, Northern↗

PubMed1998

Expression of four CEA family antigens (CEA, NCA, BGP and CGM2) in normal and cancerous gastric epithelial cells: up-regulation of BGP and CGM2 in carcinomas.

Four human carcinoembryonic antigen (CEA) family members, CEA (CD66e), non-specific cross-reacting antigen (NCA, CD66c), biliary glycoprotein (BGP, CD66a) and CEA gene-family member 2 (CGM2), are expressed in normal mucosal epithelia of the colon. Expression of BGP and CGM2 has recently been demonstrated to be down-regulated in colorectal adenocarcinomas. We have now investigated the expression of the 4 CEA family antigens in gastric adenocarcinoma and carcinoma cell lines in comparison with adjacent normal gastric mucosa. The transcripts of the CEA, NCA and BGP genes evaluated by reverse transcription-polymerase chain reaction were detectable at various levels in all the gastric adenocarcinoma cell lines tested, while CGM2 mRNA was detectable in the cell lines of poorly differentiated but not of well-differentiated carcinomas. The levels of CEA mRNA in normal gastric mucosa were variable but mostly increased in adenocarcinomas. The sparse expression of NCA observed in the normal tissues was markedly up-regulated in the carcinomas. In contrast to previous findings on normal and cancerous colonic tissues, the transcripts of CGM2 were totally undetectable and those of BGP were recognized only marginally, if at all, in normal gastric mucosa, while both messages were detected at significant levels in most of the gastric adenocarcinomas. This was confirmed by in situ hybridization. Our findings indicate that expression of the CEA family antigens, particularly that of BGP and CGM2, is differently regulated in epithelial cells of the colon and the stomach.

Adenocarcinoma↗

PubMed1998

Reliability and validity of the Motor Fitness Scale for older adults in the community.

Responses to 14 items of a questionnaire on motor fitness were collected from 990 subjects, all 65 years of age or older, living in a community. The Motor Fitness Scale was created by totalling the score for each item on the questionnaire. A second-order covariance structure analysis revealed that the Scale has a unidimensional structure with three subscales, Mobility, Strength, and Balance. The Scale appeared highly reliable with alpha = 0.92 and test-retest = 0.92. The Scale correlated with the summary physical performance score (r = 0.59). High discriminant validity, using the age and sex of the subjects, and construct validity, using their health status and level of sports participation, were confirmed for both the Scale and the summary physical performance score. The Scale can also discriminate among older persons who are at the high end of the functional spectrum that determines their level of competence in daily life. The results suggest that the Motor Fitness Scale is a feasible substitute for the application of physical performance measures in assessing the physical function of older adults in the community.

Activities of Daily Living↗

PubMed1998

Exogenous thrombospondin stimulates the proliferation of non-thrombospondin-producing cells.

The effects of thrombospondin (TSP) on the proliferation of two different human carcinoma cell lines (KIM-1 and CW-2) were investigated. The characterization of these two carcinoma cells by immunohistochemistry using anti-TSP antibody and anti-TSP-receptor antibody showed that the KIM-1 had TSP-receptors and TSP, while the CW-2 had only TSP-receptors. The addition of exogenous TSP (10 or 20 microg/ml) to culture medium stimulated the cell proliferation of CW-2 but not that of KIM-1. The cell count for CW-2 was increased dosage-dependently from 10.3 0.6x104/ml at zero TSP concentration to 12.9 0.6x104/ml at 10 microg/ml TSP concentration and to 14.7 0. 4x104/ml at 20 microg/ml TSP (each p<0.0001). In conclusion, though TSP promoted the proliferation of non-TSP-producing cells, it did not promote proliferation of TSP-producing cells. Therefore, it is predicted that TSP was already at saturated activity concentration in the TSP-producing cell line (KIM-1).

Blotting, Western↗

PubMed1998

[A case of gastric cancer with multiple bone metastasis treated by nocturnal 5-fluorouracil infusion combined with pamidronate].

Nocturnal infusion of 5-fluorouracil (5-FU) combined with pamidronate was performed in a 62-year-old male gastric cancer patient with multiple bone metastasis. The patient was administered 500 mg of 5-FU five days a week continuously for 10 hours per day from 21 o'clock to 7 o'clock for 5 months. In addition to 5-FU, 45 mg of pamidronate was administered intravenously every two weeks. Remarkable sclerotic changes were shown during the treatment in the bone metastatic foci, and the range of motion was enlarged. Serum levels of CEA and CA19-9 were decreased to the normal levels. There were no serious side effects such as myelosuppression, diarrhea or palmo-plantar dermatitis. This combination therapy of nocturnal infusion of 5-FU with pamidronate was considered effective for gastric cancer in patients with multiple bone metastasis without serious side effects.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed1996

Preparation and characterization of two human carcinoembryonic antigen family proteins of neutrophils, CD66b and c, in silkworm larvae.

As a step to investigate the cell adhesion mechanism and physiological roles of two CD66 antigens in human neutrophils, carcinoembryonic antigen gene family member 6 (CGM6, CD66b) and nonspecific cross-reacting antigen (NCA, CD66c), we prepared their soluble recombinant forms in silkworm larvae. Each cDNA fragment for CGM6 and NCA was ligated into the transfer vector pBK283 after modification to encode the protein lacking the membrane anchor. The resultant vectors were introduced to the Bombyx mori nuclear polyhedrosis virus, with which silkworm larvae were infected. Recombinant proteins secreted into the hemolymph of larvae at concentrations up to 1.3 mg/ml were purified by cation exchange followed by gel filtration or antibody affinity chromatography. The smaller apparent masses of the antigens compared with those of the native antigens appeared to be primarily due to incomplete glycosylation. Both recombinant antigens are quite similar to the corresponding native antigens in terms of the antigenic reactivity against a panel of CD66 monoclonal antibodies. In addition, the recombinant CGM6 and NCA exhibited cell binding activity against CHO cells expressing NCA and CGM6, respectively. Thus the two biologically active recombinant CD66 antigens prepared in large quantities in silkworm larvae should be useful for their functional studies, and our present system will be available for the production and purification of other carcinoembryonic antigen family members, whose biological functions are also unknown.

Animals↗

PubMed1996

Walking patterns and finger rhythm of older adults.

Walking patterns and rhythmic movement of the fingers were examined in a total of 1,134 male and female community residents 65 years of age and over. Walking patterns were characterized according to the ratio of step length divided by step rate (cadence), called the Walk Ratio, during level walking at preferred and maximum speeds. The walking pattern tended to change according to age; older subjects walked with shorter steps (smaller Walk Ratio). Rhythmic movement was examined using the finger-tapping test in time to the sound of a 4-Hz metronome. Hastened tapping or finger festination, in which the subject tapped faster than requested (constant error of 3 msec. and over in the intertap interval), was characteristic of aging; 16.8% of the subjects exhibited finger festination and the occurrence increased with age, especially among those in their eighties (29.3%). Finger festination was accompanied by walking patterns with an increased step rate, or a smaller Walk Ratio. These characteristics of aging were discussed as similar to extrapyramidal symptoms of walking and rhythm production in patients with Parkinson's disease.

Acceleration↗

PubMed1996

[Iatrogenic injury of tracheobronchial membranous wall].

Tracheobronchial injuries remain uncommon, but they are of great significance, because they can result in death or substantinal functional compromise. Such injuries are mostly from blunt trauma and motor vehicle accidents, but there also is an incidence of penetrating thoracic trauma inclunding iatrogenic accidents. Three cases of iatrogenic injury of tracheobronchial membranous wall were reported. Two cases were tear at the membraneous portion of the left main bronchus and trachea by forceful endotracheal intubation. Another case was tracheal membranous wall injury during operation at blunt dissection for esophageal carcinoma. We reported the emergent managements for iatrogenic injury of tracheobroncheal membranous wall in differents 3 ways. We should select the best treatment according to the condition of the patients and situation of the injury.

Aged↗

PubMed1994

Non-proteolytic release of carcinoembryonic antigen from normal human colonic epithelial cells cultured in collagen gel.

Recent studies have shown that, even with a minimal content of carcinoembryonic antigen (CEA), normal human colonic epithelial cells express substantial amounts of CEA mRNA and colonic mucosal fragments cultured in vitro produce CEA quite actively, indicating that CEA should no longer be considered to be of an oncofetal nature. To understand the basis of the usefulness of CEA as a tumor marker, we analyzed the release of CEA, a glycosyl-phosphatidylinositol (GPI)-anchored protein, from colonic epithelial cells, by culturing isolated colonic crypts in collagen gel. The crypts appeared to preserve their morphological and biochemical integrity in the gel for at least 16 hr, and released CEA spontaneously. Three forms of CEA--spontaneously released CEA, CEA liberated with phosphatidylinositol-specific phospholipase C (PI-PLC) and CEA in cell lysates--were indistinguishable on SDS-PAGE. This is in contrast to recombinant CEA spontaneously released from CHO transfectants, which showed a smaller molecular mass than that of PI-PLC-cleaved recombinant CEA. By phase separation using Triton X-114, CEA in the cell lysates of crypts was separated mostly into the detergent phase, while the spontaneously released and the PI-PLC-cleaved CEA were separated into the aqueous phase. When the cells were metabolically labeled with the precursors of the GPI-anchor, 3H-ethanolamine but not 3H-palmitic acid was found in the spontaneously released CEA. These findings suggest that, in contrast to the proteolysis-like release of the recombinant CEA from CHO cells, CEA in normal colonic epithelial cells is released by a non-proteolytic cleavage, which probably occurs through the action of some endogenous phospholipase.

Animals↗

PubMed1994

[Age-related difficulty in rhythmic movement].

In order to identify a characteristic difficulty in rhythmic movements with aging, a total of 380 healthy participants aged from 18 to 85 years (group 1), and 1,134 elderly community residents aged from 65 to 89 years (group 2), were examined using a finger-tapping test. The test requested the participant to tap in time to a periodic sound train with frequencies of 2, 3, 4, and 5 Hz (cycles/sec) for group 1, and with 4 Hz for group 2. Tapping deviated towards a faster rate from the stimulus frequency by more than 3 msec at 4 Hz and/or 5 Hz, "hastened tap" (HT), was found to be characteristic of aging. In group 1, the participants who exhibited HT increased with age and reached more than 35% in their 60s and 70s. In group 2, the percentages of participants with HT at 4 Hz were 14.6 (60s), 15.9 (70s) and 29.3 (80s), which were very close to the 16.9% of participants with HT at 4 Hz over 65 years in group 1. This figure suggested that more than 50% of participants over 80 years exhibited HT in tapping test at 2 through 5 Hz. HT in the elderly appears to be similar to hastened tapping observed typically in patients with Parkinson's disease, suggesting a parallel of extrapyramidal motor dysfunction between normal aging and parkinsonism.

Adolescent↗

PubMed1994

[Effect of aging on the aerobic capacity measured by a step-test].

The purpose of this study was to develop a step-test in order to evaluate age-related change in aerobic capacity. A total of 149 healthy men of age through 18 to 83 yrs ascended and descended a single step of 0.2 m height in time to a metronome. The step rate increased step-wise through three stages, each of 3 min duration; 15, 20 and 25 (step/min) for subjects aged 59 or less, and 10, 15, and 20 (step/min) for those age 60 and over. Using the linear relationship between load and heart rate, physical work capacity (PWC, watt/kg) was estimated as the work load with maximum heart rate predicted by age (220-age). In the elderly group (n = 34), heart rate at the end of the last stage was within 60-80% of the maximum heart rate for 25 subjects, and more than 80% for the other 9 subjects. No risky arrhythmia or significant ST change on ECG appeared in any subject. A retest of the step-test for 16 elderly subjects showed its repeatability and a linear relationship between heart rate and oxygen consumption. The results suggested that the step-test was applicable to the elderly in regard to appropriate work load and safety. The PWC significantly declined with aging (r = 0.52, p < 0.001). Relative aerobic capacity, taking that at age 20 as 100%, was 60% and 53% for subjects aged 60 and 70, respectively, which was in good agreement with available reports which measured oxygen consumption directly.

Adolescent↗

PubMed1992

Three different NCA species, CGM6/CD67, NCA-95, and NCA-90, are comprised in the major 90 to 100-kDa band of granulocyte NCA detectable upon SDS-polyacrylamide gel electrophoresis.

Human granulocytes express several species of nonspecific cross-reacting antigens (NCA), glycoproteins belonging to the carcinoembryonic antigen (CEA) family. Our previous studies have shown that at least two different NCA of 95 and 90 kDa are contained in the major NCA band of 90 to 100 kDa detectable upon gel electrophoresis of immunoprecipitates obtained from the cell surfaces of granulocytes with polyclonal anti-NCA. In the present study, the 90 to 100-kDa NCA band was found to include one more species of 100 kDa. This component was reactive with an anti-CD67 antibody as well as polyclonal anti-NCA and released from the cell surface with phosphatidylinositol-specific phospholipase C, indicating that the 100-kDa NCA species is CD67. Both antibodies revealed high binding activities with a recombinant protein of CGM6, which has been identified in a leukocyte cDNA library as an NCA gene and found to encode a glycosyl-phosphatidylinositol-anchored heterotypic cell adhesion molecule. Furthermore, the apparent molecular mass of the deglycosylated CD67 (38 kDa) corresponded with that of the CGM6 protein. These results suggest that CD67 is equivalent to the NCA species CGM6.

Animals↗

PubMed1992

Augmented expression and release of nonspecific cross-reacting antigens (NCAs), members of the CEA family, by human neutrophils during cell activation.

Nonspecific cross-reacting antigens (NCAs) are a group of human glycoproteins immunologically cross-reactive with carcinoembryonic antigen (CEA). Our previous studies have shown that at least seven NCA glycoproteins different in molecular weight and antigenic reactivity, including a species corresponding to CD67, can be detected in neutrophil granulocytes. In the present paper, it is demonstrated that neutrophil activation induced with soluble stimulators, the calcium ionophore A23187, N-formylmethionyl-leucyl-phenylalanine, and phorbol myristate acetate, results in augmented release and cell surface expression of NCAs. The NCA release was correlated with the discharge of azurophil granules but not with that of specific granules and was attributable to the release of NCA species of 53 and 30 kd. The increased NCA expression on the cell surface was due to increments of the NCAs of 160, 100 (CD67), 95, 90, 30, and 26 kd. These results, together with the previous findings that the CEA family members can mediate intercellular adhesion and bind Escherichia coli in vitro, imply that the neutrophil NCAs participate in the functions of neutrophils such as phagocytosis, chemotaxis, and adherence.

Antigens, Neoplasm↗

PubMed1991

Inhibitory effect of dietary iron deficiency on the induction of putative preneoplastic foci in rat liver initiated with diethylnitrosamine and promoted by phenobarbital.

The effects of dietary iron deficiency on induction of putative preneoplastic, gamma-glutamyltransferase (GGT)-positive hepatocyte focal lesions in the liver of rats treated with diethylnitrosamine (DEN) followed by phenobarbital (PB) were investigated. Male Fischer 344 rats of 4 weeks old were placed on an iron deficient (ID) diet containing less than 5 p.p.m. of iron or an iron supplemented (IS) diet containing 180 p.p.m. of iron throughout experimental period of 12 weeks. Both groups of rats were administered 200 mg kg-1 body weight of DEN by a single intraperitoneal injection at Week 4 followed by PB mixed into each diet at a concentration of 0.05% from Week 6 to the final sacrifice at Week 12 when induction of GGT-positive foci was quantitatively analysed. On the ID and IS diets, respective numbers of GGT-positive foci were 6.3 and 14.2 cm-2. The sizes of foci were not altered by the iron content of the diet. The present results indicate that iron plays a role in the development of preneoplastic foci in the livers of rats initiated with DEN and promoted by PB especially in the initiation phase.

Anemia, Hypochromic↗

PubMed1990

Endocytosis-independent uptake of liposome-encapsulated superoxide dismutase prevents the killing of cultured hepatocytes by tert-butyl hydroperoxide.

Liposome-encapsulated (LSOD) or free (FSOD), human recombinant Cu-Zn superoxide dismutase prevented the killing of cultured rat hepatocytes by tert-butyl hydroperoxide (TBHP). A dose of 32 U/ml of LSOD reduced the cell killing by 50%. By contrast, it required 288 U/ml of FSOD to similarly reduce the toxicity of TBHP by 50%. Both LSOD and FSOD increased the cell-associated superoxide dismutase activity of the cultured hepatocytes. Whereas 64 U/ml of LSOD increased cell-associated superoxide dismutase activity fourfold, it required 500 U/ml of FSOD to achieve a similar increase. Furthermore, methylamine, benzyl alcohol, cytochalasin B, oligomycin, and monensin, all inhibitors of endocytosis, prevented the increase in cell-associated superoxide dismutase produced by 500 U/ml of FSOD. These same inhibitors had no effect on the increase in cell-associated superoxide dismutase activity produced by a much lower concentration of LSOD. Thus, liposome-encapsulated superoxide dismutase prevented the cell killing by TBHP more efficiently than free superoxide dismutase because it more efficiently entered the hepatocytes by a mechanism that was independent of the endocytosis responsible for the uptake of FSOD. These data further define the conditions of the toxicity of TBHP. The target hepatocyte must contribute superoxide anions, in addition to the previously shown ferric iron. It is hypothesized that superoxide anions reduce ferric to ferrous iron; the latter then reacts with the hydroperoxide to form tert-butyl alkoxyl radicals. Such radicals are potent oxidizing agents that can initiate the peroxidation of cellular lipids previously shown to lethally injure the hepatocytes.

Animals↗

PubMed1990

Production of both 8-hydroxydeoxyguanosine in liver DNA and gamma-glutamyltransferase-positive hepatocellular lesions in rats given a choline-deficient, L-amino acid-defined diet.

The comparative carcinogenic activities of a choline-deficient, L-amino acid-defined diet (CDAADD) and a purified choline-deficient diet (CDD) for rat liver were studied in terms of both 8-hydroxydeoxyguanosine induction, a marker of DNA damage induced by oxidative stress, and development of gamma-glutamyltransferase (GGT)-positive putative preneoplastic lesions, including foci and hyperplastic nodules. Twelve weeks after the beginning of treatment, DNA damage could be detected in the liver DNA of rats receiving either CDAADD or CDD, the degree being significantly greater in the former case. Similarly, while GGT-positive liver lesions were induced by both CDAADD and CDD, the numbers were higher and the areas of lesions were larger in rats receiving CDAADD than in those given CDD. Histologically, hyperplastic nodules were induced in the livers of animals administered CDAADD whereas only foci were seen in the CDD case. The results thus indicate that oxidative stress might be directly involved in rat liver carcinogenesis by CDD and, to a greater degree, with CDAADD.

8-Hydroxy-2'-Deoxyguanosine↗