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Biomedical subjects

S Funamoto

Publications and source records attributed to S Funamoto.

13 recordsLinked to original sources
PubMed2001

Role of phosphatidylinositol 3' kinase and a downstream pleckstrin homology domain-containing protein in controlling chemotaxis in dictyostelium.

We show that cells lacking two Dictyostelium class I phosphatidylinositol (PI) 3' kinases (PI3K and pi3k1/2-null cells) or wild-type cells treated with the PI3K inhibitor LY294002 are unable to properly polarize, are very defective in the temporal, spatial, and quantitative regulation of chemoattractant-mediated filamentous (F)-actin polymerization, and chemotax very slowly. PI3K is thought to produce membrane lipid-binding sites for localization of PH domain-containing proteins. We demonstrate that in response to chemoattractants three PH domain-containing proteins do not localize to the leading edge in pi3k1/2-null cells, and the translocation is blocked in wild-type cells by LY294002. Cells lacking one of these proteins, phdA-null cells, exhibit defects in the level and kinetics of actin polymerization at the leading edge and have chemotaxis phenotypes that are distinct from those described previously for protein kinase B (PKB) (pkbA)-null cells. Phenotypes of PhdA-dominant interfering mutations suggest that PhdA is an adaptor protein that regulates F-actin localization in response to chemoattractants and links PI3K to the control of F-actin polymerization at the leading edge during pseudopod formation. We suggest that PKB and PhdA lie downstream from PI3K and control different downstream effector pathways that are essential for proper chemotaxis.

Actins↗
PubMed2001

An SH2-domain-containing kinase negatively regulates the phosphatidylinositol-3 kinase pathway.

SHK1 is a novel dual-specificity kinase that contains an SH2 domain in its C-terminal region. We demonstrate that SHK1 is required for proper chemotaxis and phagocytosis. Mutant shk1 null cells lack polarity, move very slowly, and exhibit an elevated and temporally extended chemoattractant-mediated activation of the kinase Akt/PKB. GFP fusions of the PH domain of Akt/PKB or the PH-domain-containing protein CRAC, which become transiently associated with the plasma membrane after a global stimulation with a chemoattractant, remain associated with the plasma membrane for an extended period of time in shk1 null cells. These results suggest that SHK1 is a negative regulator of the PI3K (phosphatidylinositol-3 kinase) pathway. Furthermore, when a chemoattractant gradient is applied to a wild-type cell, these PH-domain-containing proteins and the F-actin-binding protein coronin localize to its leading edge, but in an shk1 null cell they become randomly associated with the plasma membrane and cortex, irrespective of the direction of the chemoattractant gradient, suggesting that SHK1 is required for the proper spatiotemporal control of F-actin levels in chemotaxing cells. Consistent with such functions, SHK1 is localized at the plasma membrane/cortex, and we show that its SH2 domain is required for this localization and the proper function of SHK1.

Actins↗
PubMed2001

Signaling pathways controlling cell polarity and chemotaxis.

Many important biological processes, including chemotaxis (directional cell movement up a chemoattractant gradient), require a clearly established cell polarity and the ability of the cell to respond to a directional signal. Recent advances using Dictyostelium cells and mammalian leukocytes have provided insights into the biochemical and molecular pathways that control chemotaxis. Phosphoinositide 3-kinase plays a central and possibly pivotal role in establishing and maintaining cell polarity by regulating the subcellular localization and activation of downstream effectors that are essential for regulating cell polarity and proper chemotaxis. This review outlines our present understanding of these pathways.

Actins↗
PubMed2001

Stable chromosome aberrations in atomic bomb survivors: results from 25 years of investigation.

Frequencies of stable chromosome aberrations from more than 3,000 atomic bomb survivors were used to examine the nature of the radiation dose response. The end point was the proportion of cells with at least one translocation or inversion detected in Giemsa-stained cultures of approximately 100 lymphocytes per person. The statistical methods allow for both imprecision of individual dose estimates and extra-binomial variation. A highly significant and nonlinear dose response was seen. The shape of the dose response was concave upward for doses below 1.5 Sv but exhibited some leveling off at higher doses. This curvature was similar for the two cities, with a crossover dose (i.e. the ratio of the linear coefficient to the quadratic coefficient) of 1.7 Sv (95% CI 0.9, 4). The low-dose slopes for the two cities differed significantly: 6.6% per Sv (95% CI 5.5, 8.4) in Hiroshima and 3.7% (95% CI 2.6, 4.9) in Nagasaki. This difference was reduced considerably, but not eliminated, when the comparison was limited to people who were exposed in houses or tenements. Nagasaki survivors exposed in factories, as well as people in either city who were outside with little or no shielding, had a lower dose response than those exposed in houses. This suggests that doses for Nagasaki factory worker survivors may be overestimated by the DS86, apparently by about 60%. Even though factory workers constitute about 20% of Nagasaki survivors with dose estimates in the range of 0.5 to 2 Sv, calculations indicate that the dosimetry problems for these people have little impact on cancer risk estimates for Nagasaki.

Adolescent↗
PubMed1996

Antisense RNA inactivation of gene expression of a cell-cell adhesion protein (gp64) in the cellular slime mold Polysphondylium pallidum.

The gp64 protein of Polysphondylium pallidum has been shown to mediate EDTA-stable cell-cell adhesion. To explore the functional role of gp64, we made an antisense RNA expression construct designed to prevent the gene expression of gp64; the construct was introduced into P. pallidum cells and the transformants were characterised. The antisense RNA-expressing clone L3mc2 which had just been harvested at the growth phase tended to re-form in aggregates smaller in size than did the parental cells in either the presence or absence of 10 mM EDTA. In contrast, 6.5-hour starved L3mc2 cells remained considerably dissociated from each other after 5 minutes gyrating, although aggregation gradually increased by 50% during a further 55 minutes gyrating in the presence of 10 mM EDTA. Correspondingly, L3mc2 lacked specifically the cell-cell adhesion protein, gp64. We therefore conclude that the gp64 protein is involved in forming the EDTA-resistant cell-cell contact. In spite of the absence of gp64, L3mc2 exhibited normal developmental processes, a fact which demonstrates that another cell-cell adhesion system exists in the development of Polysphondylium. This is the first report in which an antisense RNA technique was successfully applied to Polysphondylium.

Cell Adhesion Molecules↗
PubMed1995

Further observations on abnormal brain development caused by prenatal A-bomb exposure to ionizing radiation.

The pervasiveness of abnormal brain development caused by prenatal exposure to ionizing radiation is still largely unknown. The relationship between A-bomb radiation dose and two measures of neuromuscular performance, one of grip strength and the other of the fine motor coordination required in repetitive action, is described. A multivariate analysis of covariance was used to evaluate the effect of several covariates, such as prenatal radiation exposure and some physical measurements or IQ adding city and sex as categorical factors. When mentally retarded cases were included, a statistically significant effect of radiation exposure on the grip strength and repetitive-action test scores was seen in the 8-15-week postovulation period, and a statistically suggestive effect at 16-25 weeks postovulation. No effect of radiation exposure on the two test scores was noted for prenatal exposure in either of the aforementioned periods when mentally retarded cases were excluded, but a statistically significant diminution of IQ was noted for exposures > or = 16 weeks postovulation. We discuss, from the biological perspective, the projected standard scores for exposures > or = 16 weeks postovulation, and the possibility of lower IQ, small head and mild mental retardation related to radiation exposures < or = 15 weeks postovulation with mentally retarded cases excluded.

Adolescent↗
PubMed1995

Height reduction among prenatally exposed atomic-bomb survivors: a longitudinal study of growth.

Using a random coefficient regression model, sex-specific longitudinal analyses of height were made on 801 (392 male and 409 female) atomic-bomb survivors exposed in utero to detect dose effects on standing height. The data set resulted from repeated measurements of standing height of adolescents (age 10-18 y). The dose effect, if any, was assumed to be linear. Gestational ages at the time of radiation exposure were divided into trimesters. Since an earlier longitudinal data analysis has demonstrated radiation effects on height, the emphasis in this paper is on the interaction between dose and gestational age at exposure and radiation effects on the age of occurrence of the adolescent growth spurt. For males, a cubic polynomial growth-curve model applied to the data was affected significantly by radiation. The dose by trimester interaction effect was not significant. The onset of adolescent growth spurt was estimated at about 13 y at 0 Gy. There was no effect of radiation on the adolescent growth spurt. For females, a quadratic polynomial growth-curve model was fitted to the data. The dose effect was significant, while the dose by trimester interaction was again not significant.

Adolescent↗
PubMed1994

Evidence of radiation-induced reduction of height and body weight from repeated measurements of adults exposed in childhood to the atomic bombs.

Reduction of growth from exposure to atomic bomb radiation has been examined using individuals under 10 years old at the time of the bombing (ATB) and a growth curve analysis based on measurements of height and weight made in the course of the 4th-7th cycles of the Adult Health Study examinations (1964-1972). As expected, the largest difference in growth to emerge is between males and females. However, a highly significant reduction of growth associated with dose (DS86) was observed among those survivors for whom four repeated measurements of height and weight were available. Longitudinal analysis of a more extended data set (n = 821), using expected values based on simple linear regression models fitted to the three available sets of measurements of height and weight on the 254 individuals with a missing measurement, also indicates a significant radiation-related growth reduction. The possible contribution of such factors as poor nutrition and disruption of normal family life in the years immediately after the war is difficult to evaluate, but the effects of socioeconomic factors on the analysis of these data are discussed.

Adult↗
PubMed1991

Tumour necrosis factor-alpha immunodetection in blood monocytes and serum: preliminary findings in weight-losing cancer patients.

The peptide tumour necrosis factor-alpha (TNF-alpha) is a central mediator of the host response. Identifying where and when TNF-alpha is produced may give insights into its potential role in various pathophysiological states. This paper describes a quantitative analysis of TNF-alpha expression in peripheral blood monocytes (PBM) at the single cell level. A pilot study has been undertaken, using this method to assess TNF-alpha expression in PBM from healthy volunteers and cancer patients. We also report mildly elevated serum TNF-alpha levels in the cancer patients, using an immunoradiometric assay (IRMA) sensitive to 1 pg/mL of recombinant TNF-alpha. The results of this preliminary investigation suggest that TNF-alpha production may be altered in cancer patients.

Humans↗
PubMed1989

In vivo immunohistochemical identification of tumor necrosis factor/cachectin in human lymphoid tissue.

To find an alternative approach to the in vivo detection of tumor necrosis factor/cachectin (TNF alpha), an immunohistochemical method to identify TNF alpha in histologic sections was developed. This method employs the streptavidin-biotin immunoperoxidase technique, and TNF alpha-specific monoclonal and polyclonal antibodies, on cryostat sections of fresh frozen human lymphoid tissue. Staining was evident in most specimens displaying follicular hyperplasia, but was absent from histologically normal tissue. Both tingible body macrophages and follicular dendritic reticulum cells appeared from phenotype analysis in serial sections and by double staining experiments to constitute the main source of TNF alpha. This technique complements other systemically oriented assays that may fail to detect significant in vivo TNF alpha production and activity at a cellular level.

Humans↗