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Biomedical subjects

S Takano

Publications and source records attributed to S Takano.

At least 73 records · Page 4Linked to original sources

Evidence for stanniocalcin gene expression in mammalian bone.

Stanniocalcin (STC) acts as a regulator of calcium and phosphate homeostasis in an endocrine manner in bony fish. Recently, complementary DNAs encoding human and mouse STC have been characterized, and the messenger RNA (mRNA) expression was identified in various tissues, such as kidney, small intestine, prostate, thyroid, and ovary. Because previous studies concerning the effects of fish STC on mammalian bone have been discussed, there is a good possibility that mammalian STC is a local factor in bone. Here, we demonstrated STC mRNA expression in neonatal mouse calvaria, the primary cultured mouse osteoblast-rich fractions, and human and mouse osteoblastic cell lines. We also mapped the cellular distribution of the STC mRNA in femur and calvaria in developing mice. Several transcripts with a major 4-kb band were detected in all samples. The cellular distribution of the mRNA expression corresponded closely to osteoblasts in both femur and calvaria. Significant labeling of the STC mRNA was also identified in chondrocytes but not in osteoclasts and other bone marrow elements. These results are the first evidence that hormone may be actually expressed in osteoblasts and chondrocytes, and they strongly implicate the involvement of local STC in both endochondral and membrane bone as an autocrine/paracrine factor.

Aging↗

No-touch isolation technique reduces intraoperative shedding of tumor cells into the portal vein during resection of colorectal cancer.

BACKGROUND: The mutant-allele-specific amplification (MASA) method is capable of detecting 1 genetically altered tumor cell among thousands of normal cells. The MASA enabled us to detect occult tumor cells undetectable by histopathologic examination of lymph nodes and blood samples. METHODS: To investigate whether tumor manipulation during operation enhances cancer cell dissemination into the portal vein with use of MASA and to assess the effect of the no-touch isolation technique in the treatment of colorectal cancers, 27 colorectal cancers (17 were operated on conventionally and 10 were operated on according to the no-touch isolation technique) were screened for mutations in K-ras or p53. We next examined blood samples of the portal vein collected before, during, and after manipulation of tumors, using MASA to look for the specific mutation found in the primary tumors. RESULTS: Somatic mutations were identified in 18 of these primary tumors (11 were in the conventional resection technique group and 7 were in the no-touch isolation technique group). In 8 of 11 (73%) conventional resection technique cases, we identified the same genetic alteration of the primary tumor in the portal blood during operation, whereas only 1 patient (14%) in the no-touch isolation technique group had a positive result. CONCLUSIONS: The no-touch isolation technique may be useful to prevent cancer cells from being shed into the portal vein during surgical manipulation.

Alleles↗

Inactivation of tumor suppressor p53 by mot-2, a hsp70 family member.

The mortalin genes, mot-1 and mot-2, are hsp70 family members that were originally cloned from normal and immortal murine cells, respectively. Their proteins differ by only two amino acid residues but exhibit different subcellular localizations, arise from two distinct genes, and have contrasting biological activities. We report here that the two proteins also differ in their interactions with the tumor suppressor protein p53. The pancytosolic mot-1 protein in normal cells did not show colocalization with p53; in contrast, nonpancytosolic mot-2 and p53 overlapped significantly in immortal cells. Transfection of mot-2 but not mot-1 resulted in the repression of p53-mediated transactivation in p53-responsive reporter assays. Inactivation of p53 by mot-2 was supported by the down-regulation of p53-responsive genes p21(WAF-1) and mdm-2 in mot-2-transfected cells only. Furthermore, NIH 3T3 cells transfected with expression plasmid encoding green fluorescent protein-tagged mot-2 but not mot-1 showed an abrogation of nuclear translocation of wild-type p53. These results demonstrate a novel mechanism of p53 inactivation by mot-2 protein.

3T3 Cells↗

Malignant transformation of NIH3T3 cells by overexpression of mot-2 protein.

The murine mortalin genes, mot-1 and mot-2, are members of the hsp70 family of proteins and differ from each other by only two amino acid residues. Mot-1 is expressed in normal cells and has pancytosolic cellular distribution whereas mot-2 is found in the perinuclear region of immortal cells. We report here that a high level of expression of mot-2 protein resulted in malignant transformation of cells as analysed by anchorage independent growth and nude mice assays. A high level of protein expression is attributed to the 900 bp 3' untranslated region of the cDNA which does not have any transforming activity per se. Mortalin cDNA clones isolated from human transformed cells were also found to have transforming activity in similar assays and a high level of expression was apparent in some of the human immortalized cells that showed non-pancytosolic mortalin immunofluorescence. Taken together, the data suggest that nonpancytosolic mortalin may have a role in tumorigenesis.

3T3 Cells↗

Expression of 5-lipoxygenase and 5-lipoxygenase-activating protein mRNAs in the peripheral blood leukocytes of asthmatics.

Leukotrienes are a family of arachidonic acid metabolites with potent biological activities such as bronchoconstriction and leukocyte chemotaxis. Recent evidence has demonstrated that the 5-lipoxygenase (5-LO) and 5-lipoxygenase-activating protein (FALP) products of arachidonic acid metabolism, leukotriene C4, D4 and E4, were increased in the serum and the urine of asthmatic patients. Therefore, we examined the expression of 5-LO and FLAP mRNAs in the peripheral blood leukocytes (PBL) of 10 asthmatics and 10 controls. Both 5-LO and FLAP mRNAs of PBL in the asthmatic group were found to be significantly increased compared with those in the control group. These data suggest that up-regulation of 5-LO and FLAP mRNAs might be involved in the increased leukotriene synthesis and play an important role in the pathogenesis of asthma.

5-Lipoxygenase-Activating Proteins↗

The ND Radical: Laboratory Measurement of the N = 2-1 Rotational Transition at 1 THz.

The N = 2-1 pure rotational transition of the ND radical (X3Sigma-) near 1 THz was measured with the Cologne terahertz spectrometer. The ND radical was produced in a dc discharge of a flowing mixture of deuterated ammonia and helium. Frequencies of five strong fine-structure transitions with associated hyperfine components were precisely measured and analyzed to determine a complete set of accurate molecular constants, e.g., the rotational and centrifugal distortion constants B0 = 263265.4735(45) MHz and D0 = 14.62876(74) MHz, together with the fine and hyperfine constants. Based on these constants, the rotational frequencies of ND up to 4 THz were predicted with the aim to guide future astronomical searches. For completeness we also include similar predictions for NH. Copyright 1998 Academic Press.

Journal Article↗

Strain-differences in proliferative activity of uterine endometrial cells in Donryu and Fischer 344 rats.

Strain differences in proliferative activities of uterine endometrial cells were sequentially investigated in Donryu and Fischer 344 (F344) rats until 52 weeks of age. At 13 and 26 weeks of age, when all rats showed a normal estrous cycle, histological findings for the endometrium at each estrous stage were similar in both strains. Mitoses of epithelial cells were frequently observed during the proestrous and metestrous stages, and especially in the latter, but were very few in the estrous stage. The bromodeoxyuridine(BrdU)-labeling indices coincided with the histological findings, the results indicating a peak of cell proliferative activity in the metestrous stage. In Donryu rats, estrous cycle abnormalities increased age-dependently after 26 weeks of age, and almost all animals showed persistent estrus at 52 weeks of age. At 39 and 52 weeks of age, the rats showing persistent estrus displayed similar vaginal smears and histological findings to those of younger rats in the estrous stage, but the BrdU-labeling indices of epithelial cells were age-dependently increased. In contrast, vaginal smear of F344 rats indicated a normal estrous cycle at all ages, with no changes in BrdU-labeling being observed. Previously, we reported that the Donryu rat is a high incidence strain in terms of spontaneous development of endometrial adenocarcinomas, this being related to an age-dependent hormonal imbalance, characterized in particular by an increased estrogen/progesterone ratio. The F344 rat in contrast is a low incidence strain. The present results thus indicate that a constant high-level of proliferative activity of epithelial cells may play an important role, together with an age-related hormonal imbalance, in the spontaneous development of endometrial carcinomas in Donryu rats.

Adenocarcinoma↗

The use of BRM-activated killer cells in adoptive immunotherapy: a pilot study with nine advanced cancer patients.

Adoptive immunotherapy using MHC-nonrestricted-lymphocytes, peripheral blood gammadelta T cells and NK cells was devised. Peripheral blood mononuclear cells (3 x 10(7)) were selected by immobilization to anti-CD3 monoclonal antibody for 4 days and cultured for 2 weeks in the presence of IL-2. Thereafter they were reactivated by 500 U/ml of IFN-alpha and 1000 U/ml of IL-2 for 1 hour. Enhancement of NK and LAK activities was confirmed. Peripheral blood gammadelta T cells proliferated in response to immobilized anti-CD3 antibody (3% to 30%). Approximately 6 x 10(9) BRM-activated killer (BAK) cells composed of CD56+ gammadelta T cells and CD56+ NK cells, were dispensed to cancer patients via intravenous drip infusion. Nine patients were treated with BAK cells every 2 weeks or every month on an outpatient basis. During the course of adoptive immunotherapy, the crossed affinity immunoelectrophoresis (CAIE) pattern of serum immunosuppressive acidic protein (IAP) was analysed. Both the production and glycosylation pattern of IAP is changed in response to tumor enlargement and may therefore act as a marker of the disease progression. During the course of BAK therapy, the glycosylation IAP pattern of 6 patients changed from tumor (T) to normal (N). In addition, the performance status of all patients was maintained at 90-100% of the Karnofsky scale and any side effects including fever were not observed during treatments with BAK cells. Moreover, the overall quality of life (QOL) of the patients, scored at the Face scale was favorable. In addition, blood levels of activated gammadelta T cells producing IFN-gamma were assayed as an indication marker of BAK therapy. The normal range of IFN-gamma producing gammadelta T cells comprised 6.9 +/- 0.9% of peripheral blood mononuclear cells (PBMC), according to a single cell FACScan analyses of PBMCs derived from normal individuals. IFN-gamma producing gammadelta T cells of Patients No. 8 and 9, who received extensive chemotherapy before initiation of BAK therapy, comprised only 0.2% and 2% of PBMC, respectively. These patients died 3 and 6 months after beginning BAK therapy. Peripheral blood gammadelta T cells of Patients Nos. 1-7 proliferated in response to immobilized anti-CD3 antibody and the frequency of IFN-gamma producing gammadelta T cells in PBMC preparation of these patients were over 3% before initiation of BAK therapy. Since our data show a positive correlation between survival time and initial gammadelta T cell counts, a low frequency of these cells may contraindicate BAK therapy.

Adult↗

Regional differences in the effect of exercise intensity on thermoregulatory sweating and cutaneous vasodilation.

To investigate regional body differences in the effect of exercise intensity on the thermoregulatory sweating response, nine healthy male subjects (23.2 +/- 0.4 year) cycled at 35, 50 and 65% of their maximal O2 uptake (VO2max) for 30 min at an ambient temperature of 28.3 +/- 0.2 degrees C and a relative humidity of 42.6 +/- 2.4%. Local sweating rate (msw) on the forehead, chest, back, forearm and thigh increased significantly with increases in the exercise intensity from 35 to 50% VO2max and from 50 to 65% VO2max (P < 0.05). The mean values for the density of activated sweat glands (ASG) at 50 and 65% VO2max at the five sites were significantly greater than at 35% VO2max. The mean value of the sweat output per gland (SGO) also increased significantly with the increase in exercise intensity (P < 0.05). The patterns of changes in ASG and SGO with an increase in exercise intensity differed from one region of the body to another. Although esophageal temperature (Tes) threshold for the onset of sweating at each site was not altered by exercise intensity, the sensitivity of the sweating response on the forehead increased significantly from 35 to 50 and 65% VO2max (P < 0.05). The threshold for cutaneous vasodilation tend to increase with exercise intensity, although the exercise intensity did not affect the sensitivity (the slope in the relationship Tes vs. percentage of the maximal skin blood flow) at each site. Tes threshold for cutaneous vasodilation on the forearm was significantly higher at 65% VO2max than at either 35 or 50% VO2max, but this was not observed at the other sites, such as on the forehead and chest. These results suggest that the increase in msw seen with an increasing intensity of exercise depends first on ASG, and then on SGO, and the dependence of ASG and SGO on the increase in msw differs for different body sites. In addition, there are regional differences in the Tes threshold for vasodilation in response to an increase in exercise intensity.

Adult↗

Replication error frequencies in primary hepatocellular carcinoma: a comparison of solitary primary versus multiple primary cancers.

AIMS/BACKGROUND: Replication errors (RERs) at microsatellite loci are associated with the development of not only hereditary nonpolyposis colon cancer but also sporadic cancers. To examine the association between RERs and human hepatocarcinogenesis, we looked for microsatellite instability in solitary and multiple primary hepatocellular carcinomas (HCCs). METHODS: DNAs were extracted from 34 solitary primary HCCs and 14 HCCs with multiple primary cancers. Twelve microsatellite alleles were amplified by PCR from the DNAs, and RERs were assessed by their mobility shift. RESULTS: RERs were found in only two cases (6%) of solitary HCC and four cases (29%) of HCC with multiple primary cancers. Two of the four HCCs with multiple primary cancers showed RERs at two and three microsatellite loci, respectively. Of 12 microsatellite loci examined, TP53 and D16S476 sensitively detected RERs in HCCs with multiple primary cancers, at a frequency of 23% and 33%, respectively. CONCLUSIONS: RERs are rarely associated with carcinogenesis in human primary HCC, and RER-related genetic alterations may be associated with a part of HCC with multiple primary cancers. If future studies confirm this association, then the two probes TP53 and D16S476 may be useful for the prediction of development of multiple primary cancers with HCC.

Adolescent↗

Preparation of a monoclonal antibody specific for human stanniocalcin.

Stanniocalcin (STC) is a glycoprotein hormone that was first identified in fish, where it regulates the calcium level in the body fluid. The cDNA which encodes human STC has recently been reported but the function has not been completely elucidated. We have prepared a monoclonal antibody against human STC using an analogous peptide of the putative antigenic domain in human STC; it was conjugated with keyhole limpet hemocyanin (KLH). The monoclonal antibody specifically stained the distal convoluted tubules in human kidney which is a putative target organ of STC. The ELISA was established using the monoclonal antibody and recombinant human STC as a standard antigen. The monoclonal antibody prepared in this study provides a useful tool for clinical studies of STC in human.

Amino Acid Sequence↗

Control of circadian variation in skin blood flow response to heat stress.

Six male subjects had their lower legs immersed in water at 42 degrees C for 60 min at 4 different times of the day to study whether the skin blood flow response to passive heat stress shows circadian variation in the relationship between skin blood flow and local sweating rate. When skin blood flow was plotted against local sweating rate, three distinct phases were observed. Phase A, an increase in skin blood flow without sweating was maximal at night. But the slope of the regression line did not change over the day in Phase C. These findings suggest that there is circadian variation in the skin blood flow response before onset of sweating during passive heat stress. This variation might be related, in part, to the circadian rhythm in vasoconstrictor activity.

Adult↗

Facial spasm and paroxysmal tinnitus associated with an arachnoid cyst of the cerebellopontine angle--case report.

A 59-year-old female presented with a 3-year history of hemifacial spasm and paroxysmal tinnitus associated with an arachnoid cyst of the cerebellopontine angle, which was identified preoperatively by magnetic resonance imaging. Surgical decompression of the cyst and arterial decompression of the facial and acoustic nerves at their root exit zones resulted in complete resolution of the symptoms.

Arachnoid Cysts↗

Basal perforating artery aneurysm within the cavum septi pellucidi. Case report.

The authors describe the case of a 16-year-old boy presenting with clinical onset of subarachnoid hemorrhage. The patient was found to have a small aneurysm arising from the distal portion of a basal perforating branch of the anterior cerebral artery (ACA), lying within a cavum septi pellucidi. Neuroimaging demonstrated a hematoma within the cavum septi pellucidi and the aneurysm was resected via a transcallosal approach guided by an intraoperative portable digital subtraction angiography (DSA) system. The origin of the aneurysm was presumed to be idiopathic. This report is the first to describe an aneurysm of a basal perforating artery arising from the ACA that resulted in an intracaval hematoma. When approaching small vascular lesions during surgery, intraoperative DSA is the method of choice to identify the precise trajectory and distance to the lesions.

Adolescent↗

[Arterial infusion of SMANCS for multiple recurrent tumors after hepatic resection].

We investigated the possible side effects and efficacy of arterial infusion of SMANCS as compared to Lipiodol + epirubicin TAE for multiple recurrent tumors after hepatic resection. As a result, no significant difference in GOT, GPT, and total bilirubin was observed between the two groups. No significant difference was found in white blood cell count and platelet count, and there was no significant difference in clinical side effects between the two groups. Grade III response rates after arterial infusion of SMANCS were found in 4 patients (66.6%), and these results showed no significant difference as compared to Lipiodol + epirubicin TAE. Proper hepatic arterial infusion of SMANCS appeared to be useful in multiple recurrent tumors from the standpoints of safety and the rate of Lipiodol deposition.

Adult↗

[Flow cytometric analysis of helper T cell subsets (Th1 and Th2) in healthy adults].

To estimate frequency of T helper (Th) 1, Th2, and other related subsets in T cells among healthy Japanese, we determined the frequencies of interferon (IFN)-gamma- and/or interleukin (IL)-4-producing cells in stimulated peripheral blood lymphocytes (PBL) from 51 healthy adults of various ages by three-color flow cytometry using FITC-labeled anti-IFN-gamma, PE-conjugated anti-IL-4, and peridinin chlorophyll protein (PerCP)-labeled anti-CD4 or anti-CD3 antibody. As a result, proportions (mean +/- SD) of IFN-gamma-single positive (conveniently called Th1), IL-4-single positive (Th2), and double-positive cells (Th0) in CD4+ PBL, and homologous cell populations in CD3+ PBL (Th1 type, Th2 type, and Th0 type cells) were 21.3 +/- 8.2%, 2.7 +/- 1.1%, 1.9 +/- 1.0%, 39.5 +/- 13.5%, 1.7 +/- 0.8%, and 1.8 +/- 1.0%, respectively. Percentages of Th1 and Th2 were roughly consistent with those reported previously. Proportions of Th1, Th2, Th1 type, and Th2 type cells were found to increase with the age, whereas there were no significant difference in sex at least in 20s. Furthermore, positive correlation was seen between Th1 and Th1 type (r = 0.79), Th2 and Th2 type (r = 0.87), and Th0 and Th0 type (r = 0.86). Frequencies of these subsets in each donor were maintained at substantially the same levels for at least 3 months, suggesting existence of particular and stable Th1/Th2 balance among healthy individuals. As reported, 6 patients with atopic dermatitis examined in this study showed significantly lower frequencies of Th1 (11.0 +/- 2.5%) as compared with healthy donors (p < 0.01), implying usefulness of the immune test.

Adult↗