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Biomedical subjects

K Yoshikawa

Publications and source records attributed to K Yoshikawa.

At least 235 records · Page 13Linked to original sources
PubMed1998

Electrophoresis of long DNA molecules in linear polyacrylamide solutions.

Electrophoresis of long DNA (T4 DNA; 166 kb, S. pombe chromosomal DNA; 3-6 Mb) in linear polyacrylamide solutions was investigated by fluorescence microscopy and capillary electrophoresis. In the past studies on electrophoresis of long DNA in a polymer solution, it was reported that DNA migrates in 'U-shape conformation'. We found that at higher polymer concentrations, the shape of the migrating DNA changes from U shape to linear shape ('I-shape conformation'). In the migration mode with the I-shape conformation, the DNA moves with almost constant velocity and constant shape. However, the migration velocity does depend on the DNA size, and it is possible to separate DNAs under this I-shape motion. Actually, Mb-sized DNAs are well separated within 5 min in the region for the I-shape motion by means of capillary electrophoresis with a DC field. Considering that it takes 20 h to separate Mb-sized DNAs by standard pulsed-field gel electrophoresis (PFGE), this results will be useful for the separation of giant DNAs.

Journal Article↗
PubMed1998

Degeneration in vivo of rat hippocampal neurons by wild-type Alzheimer amyloid precursor protein overexpressed by adenovirus-mediated gene transfer.

In an attempt to elucidate the pathological implications of intracellular accumulation of the amyloid precursor protein (APP) in postmitotic neurons in vivo, we transferred APP695 cDNA into rat hippocampal neurons by using a replication-defective adenovirus vector. We first improved the efficiency of adenovirus-mediated gene transfer into neurons in vivo by using hypertonic mannitol. When a beta-galactosidase-expressing recombinant adenovirus suspended in 1 M mannitol was injected into a dorsal hippocampal region, a number of neurons in remote areas were positively stained, presumably owing to increased retrograde transport of the virus. When an APP695-expressing adenovirus was injected into the same site, part of the infected neurons in the hippocampal formation underwent severe degeneration in a few days, whereas astrocytes near the injection site showed no apparent degeneration. These degenerating neurons accumulated different epitopes of APP, and beta/A4 protein (Abeta)-immunoreactive materials were undetected in the extracellular space. A small number of degenerating neurons showed nuclear DNA fragmentation. Electron microscopic examinations demonstrated that degenerating neurons had shrunken perikarya along with synaptic abnormalities. Microglial cells/macrophages were often found in close proximity to degenerating neurons, and in some cases they phagocytosed these neurons. These results suggest that intracellular accumulation of wild-type APP695 causes a specific type of neuronal degeneration in vivo in the absence of extracellular Abeta deposition.

Adenoviridae↗
PubMed1998

Lung as reservoir for antidepressants in pharmacokinetic drug interactions.

BACKGROUND: Although high-affinity imipramine binding sites have been reported in both rat and human lung, the role of the lungs in the pharmacokinetics of antidepressants has not received much attention. Substantial accumulation of selective serotonin-reuptake inhibitors (SSRIs) in the lungs has been reported. We have investigated the role of the lungs in pharmacokinetic drug interactions between tricyclic antidepressants and SSRIs. METHODS: We used a carbon-11-labelled form of the imipramine derivative cyanoimipramine to measure uptake in the lungs and brain of healthy volunteers by positron emission tomography. Clomipramine (50 mg) was administered to measure the effect of antidepressants with high affinity for the serotonin transporter on lung and brain uptake. FINDINGS: A large proportion of the injected 11C-cyanoimipramine (68-86% in the four volunteers) was extracted by the lungs. Clomipramine decreased the lung uptake from 68% to 35% and from 81% to 54% in the two volunteers studied. By contrast, whole-brain uptake was low in control studies (1.7-2.0% in three volunteers) and increased after clomipramine administration (to 4.5-4.9%). Plasma radioactivity was also higher after clomipramine than in control studies. INTERPRETATION: The lungs may function as a reservoir for antidepressants with high affinity to the serotonin transporter. The accumulated antidepressants may be displaced by other antidepressants, and this displacement would substantially increase plasma concentrations and thus cause toxic effects.

Adult↗
PubMed1998

Single chain observation on collapse transition in giant DNA induced by negatively-charged polymer.

Poly-glutamic acid, a polyanion, has been shown to induce the all-or-none collapse transition in individual duplex DNAs from an elongated coil into a collapsed compact state, through the observation of single chains by fluorescence microscopy. This experimental finding is discussed in relation to the change in the higher-order structure of DNAs in the cytoplasmic space induced by a change in the concentration of polyanions, such as mRNA, as a possible route for feedback in gene expression.

Bacteriophage T4↗
PubMed1998

Necdin, a postmitotic neuron-specific growth suppressor, interacts with viral transforming proteins and cellular transcription factor E2F1.

Necdin is a nuclear protein expressed in virtually all postmitotic neurons, and ectopic expression of this protein strongly suppresses the proliferation of NIH3T3 cells. Simian virus 40 large T antigen targets both p53 and the retinoblastoma protein (Rb) for cellular transformation. By analogy with the interactions of the large T antigen with these nuclear growth suppressors, we examined the ability of necdin to bind to the large T antigen. Necdin was co-immunoprecipitated with the large T antigen from the nuclear extract of necdin cDNA-transfected COS-1 cells. Yeast two-hybrid and in vitro binding analyses revealed that necdin bound to an amino-terminal region of the large T antigen, which encompasses the Rb-binding domain. Moreover, necdin bound to adenovirus E1A, another viral oncoprotein that forms a specific complex with Rb. We then examined the ability of necdin to bind to the transcription factor E2F1, a cellular Rb-binding factor involved in cell-cycle progression. Intriguingly, necdin, like Rb, bound to a carboxyl-terminal domain of E2F1, and repressed E2F-dependent transactivation in vivo. In addition, necdin suppressed the colony formation of Rb-deficient SAOS-2 osteosarcoma cells. These results suggest that necdin is a postmitotic neuron-specific growth suppressor that is functionally similar to Rb.

3T3 Cells↗
PubMed1998

Gene expression of alpha1-6 fucosyltransferase in human hepatoma tissues: a possible implication for increased fucosylation of alpha-fetoprotein.

The 1-6 fucosylated -fetoprotein (AFP) present in serum of patients with hepatocellular carcinoma (HCC) has been employed for the differential clinical diagnosis of HCC from chronic liver diseases. The molecular mechanism by which this alteration occurs, however, remains largely unknown. To address this issue, we purified GDP-L-Fuc:N-acetyl-beta-D-glucosaminide 1-6 fucosyltransferase (1-6 FucT), an enzyme involved in the 1-6 fucosylation of N-glycans from porcine brain, as well as from a human gastric cancer cell line, and cloned their genes. In this study, levels of 1-6 FucT mRNA expression and the activity of this enzyme for 12 human HCC tissues were examined and compared with that in surrounding tissues and normal livers. The mean +/- SD for 1-6 FucT activity was 78 +/- 41 pmol/h/mg in normal control liver, 202 +/- 127 pmol/h/mg in adjacent uninvolved liver tissues (chronic hepatitis: 181 +/- 106 pmol/h/mg; liver cirrhosis: 233 +/- 164 pmol/h/mg), and 195 +/- 72 pmol/h/mg in HCC tissues. The mRNA expression of 1-6 FucT was also enhanced in proportion to enzymatic activity except for a few cases, suggesting that 1-6 FucT expression is increased in chronic liver diseases, especially liver cirrhosis. Transfection of 1-6 FucT gene into cultured rat hepatocytes markedly increased 1-6 FucT activity and led to an increase in lens culinaris agglutinin (LCA) binding proteins in both cell lysates and condition media. When the 1-6 FucT gene was transfected into a human HCC cell line, Hep3B, which originally showed low levels of 1-6 FucT expression, 1-6-fucosylated AFP was dramatically increased in the condition media. Collectively, these results suggest that the enhancement of 1-6 FucT expression increased the fucosylation of several proteins, including AFP, and that the level of 1-6-fucosylated AFP in patients with HCC was in part caused by up-regulation of the 1-6 FucT gene expression.

Adult↗
PubMed1998

A novel strategy for introducing exogenous bcl-2 into neuronal cells: the Cre/loxP system-mediated activation of bcl-2 for preventing programmed cell death using recombinant adenoviruses.

We have established a novel strategy for introducing exogenous Bcl-2 into neuronal cells that is mediated by Cre/loxP recombination using recombinant adenoviral vectors. An on/off-switching cassette for Bcl-2 (CALNLbcl-2) was designed to express Bcl-2 by recombinase Cre-mediated excisional deletion of a spacer DNA flanked by a pair of loxP sites. Exogenous Bcl-2 was clearly induced in PC12 cell lines carrying CALNLbcl-2 after infection with recombinant adenovirus producing recombinase Cre (AxCANCre). Dual infection with both AxCANCre and a recombinant adenovirus bearing CALNLbcl-2 showed efficient delivery of exogenous Bcl-2 into a hybrid motoneuronal cell line and primary chicken spinal motoneurons. The delivery of foreign Bcl-2 promoted survival of motoneurons in medium either containing or lacking trophic support. Thus, this strategy for delivery of exogenous Bcl-2 will be useful for studying neuronal death as well as for introducing foreign genes into postmitotic neurons under the control of recombinase Cre.

Adenoviridae↗
PubMed1998

Inhibitory effect of lignans from the rhizomes of Coptis japonica var. dissecta on tumor necrosis factor-alpha production in lipopolysaccharide-stimulated RAW264.7 cells.

The inhibitory effect of 10 lignan constituents isolated from the rhizomes of Coptis japonica var. dissecta on tumor necrosis factor (TNF)-alpha production in lipopolysaccharide (LPS)-stimulated macrophage cell line (RAW264.7 cells) has been studied. Among them, pinoresinol, woorenoside-V and lariciresinol glycoside showed significant inhibitory activities in the range from 37% to 55% at the concentration of 25 micrograms/ml. The results are first report that the lignans isolated from Coptis japonica inhibit TNF-alpha production, and suggest that the lignan components may partly participate in antiinflammatory and antiallergic effect of Coptis japonica through the inhibition of TNF-alpha production.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1998

Estimation of deadtime in imaging human subjects.

Deadtime count loss may cause significant errors in quantitative measurements performed by scintigraphic techniques. Deadtime depends on the scattering condition, and may vary among patients. The aim of this study was to estimate deadtime in imaging a human subject. A cylindrical phantom filled with various concentrations of technetium-99m solution was imaged with a gamma camera, and deadtime was assessed assuming the gamma camera to be a non-paralysable system (multi-dose method). Deadtime for the cylindrical phantom was also estimated using a point source to monitor counting efficiency (reference source method). Radionuclide angiography of the chest was performed in 38 patients with a reference source in the field of view, and temporal changes in count loss were assessed. The deadtime for each patient was calculated by the reference source method. The deadtime for the cylindrical phantom was estimated as 6.96+/-0.09 micros by the reference source method and was almost identical to that obtained by the multi-dose method (6.93 micros). Deadtime ranged from 6.01 to 9.58 micros in radionuclide angiography, and was positively correlated with the ratio of body weight in kg to body height in cm (y=8.566 x+5.611, r=0.869). Count loss was successfully corrected using the deadtime predicted with the regression line. In summary, the deadtime in imaging a human subject has a wide range and is related to the constitution of the subject. Correction for count loss using a deadtime predicted for each patient may be helpful in improving the reliability of quantitative nuclear medicine.

Body Constitution↗
PubMed1998

The development of malignant rhabdoid tumor in a patient with Behçet's disease treated with ciclosporin.

PURPOSE: To describe a risk of malignant neoplasm in patients with Behçet's disease treated with ciclosporin. METHODS: Case report. RESULTS: A patient with Behçet's disease who was treated with ciclosporin developed a mass in his right femur. Histological examination revealed this mass was a malignant rhabdoid tumor. CONCLUSION: This is the first known case of malignant tumor developing in a patient with Behçet's disease treated with ciclosporin. This case underscores the importance of assessing possible risks in the treatment of Behçet's disease with ciclosporin.

Adult↗
PubMed1998

Diagnostic potential and pitfalls of ultrasound-guided fine-needle aspiration cytology for breast lesions.

Ultrasound (US)-guided fine-needle aspiration cytology (FNAC) is now widely accepted as a diagnostic procedure for breast lesions. Along with its advantages, US-guided FNAC also has some pitfalls. The recognition of these pitfalls for this procedure is extremely important for the strict management of the disease. We retrospectively investigated the diagnostic potential and pitfalls of US-guided FNAC in the diagnosis of breast lesions. This study consisted of 348 aspirated samples from 274 breast tumors. The rate of sufficient aspirates was 74% after a single aspiration, while sufficient materials were finally obtained from 93% of the tumors by repeated aspirations. The rate was lower in tumors measuring less than 10 mm in diameter (62%), and in sclerosing adenosis (25%). The sensitivity of FNAC was 65%, the specificity was 75%, the border diagnosis rate was 18%, and the positive predictive value was 92%. The false-negative rate was higher in noninvasive carcinoma (45%). The border diagnosis rate was also higher in scirrhous carcinoma (29%). There were also five false-positive cases. Limited to nonpalpable lesions, the sufficient aspirates rate was 70% and the accuracy was 67%.

Adenocarcinoma, Scirrhous↗
PubMed1998

Somatosensory evoked magnetic fields elicited by dorsal penile, posterior tibial and median nerve stimulation.

The aim of this study is to localize the primary sensory cortex of urogenital organs in the human brain. Using a newly developed MRI-linked magnetoencephalography system, we measured somatosensory evoked magnetic fields (SEFs) for unilateral stimuli on the dorsal penile nerve (DPN), posterior tibial nerve (PTN) and median nerve (MN). In five healthy male subjects, SEFs were clearly observed. Peak latency of the first cortical components were 63.8 +/- 9.2 ms for DPN, 39.8 +/- 3.0 ms for PTN and 20.7 +/- 0.7 ms for MN stimuli. Peak amplitude of the first cortical components were 63.1 +/- 10.8 fT for DPN, 160.2 +/- 50.1 fT for PTN and 335.2 +/- 70.3 fT for MN stimuli. Isofield map for the peak latencies indicated a single dipolar pattern for DPN as well as for PTN and MN stimuli. Using a single current dipole model, all SEF sources were localized on the contralateral central sulcus to the stimuli, indicating the primary sensory cortex. The DPN sources were localized on the interhemispheric surfaces, corresponding to previous speculations by direct cerebral stimulation. This non-invasive SEF technique promises further brain functional mapping for the urogenital organs.

Adult↗
PubMed1998

Growth inhibition of human keratinocytes by a new vitamin D3 analogue in vitro.

SM-10193 (26,26,26,27,27,27-hexafluoro-1 alpha,23(S),25-trihydroxyvitamin D3; F6-1,23(S),25-(OH)3D3) is one of the active vitamin D3 analogues, which is under exploration as a new therapeutic agent for psoriasis. In this paper, we present that SM-10193 induces the growth inhibition and differentiation of cultured normal human keratinocytes more effectively than 1,25(OH)2D3. The growth of keratinocytes was inhibited in the presence of 10(-11), 10(-10), 10(-9), 10(-8), 10(-7) and 10(-6) M of SM-10193 by 36.8, 42.1, 48.5, 47.6, 66.0 and 85.9% respectively. SM-10193 increased the number of involucrin positive cells (the marker for keratinocyte differentiation) from 4.9 +/- 0.1 to 12.5 +/- 0.8, 20.1 +/- 1.6 and 42.8 +/- 1.0% (P < 0.01) at 10(-8), 10(-7) and 10(-6) M, respectively. To elucidate the mechanism of SM-10193-induced growth inhibition, we analyzed cell cycle related distribution and the alteration of hyperphosphorylated and hypophosphorylated state of retinoblastoma protein (pRB). SM-10193 induces growth arrest in G1/G0 and G2 + M phases and an increase of the hypophosphorylated form of pRB more remarkably than 1,25(OH)2D3 does.

Calcitriol↗
PubMed1998

Biochemical and immunohistochemical analyses of keratin expression in basal cell carcinoma.

Recently we demonstrated that the keratin 17 (K17) content exceeded the K16 content in most follicular tumors, in comparison with non-follicular epithelial skin tumours by two-dimensional gel electrophoresis (2-DE), densitometry and immunohistochemistry. At present the origin of basal cell carcinoma (BCC) is unknown. So, based on the above results, we studied keratin expression in eight cases of BCC, in order to analyze tumor differentiation by both biochemical and immunohistochemical methods. Biochemically, using 2-DE and immunoblotting, stratified epithelial keratins K5/K14 and large amounts of K17 were present in all cases. Simple epithelial keratins K8 and K19 were expressed in all and half of the cases, respectively. However, hyperproliferative associated keratins (K6/K16) and keratinized keratins (K1/K10) were detected in only a few cases. Immunohistochemical studies using frozen sections with chain-specific antikeratin monoclonal antibodies against K1, K7, K8, K10, K14, K16, K17, K18 and K19 showed that BCC tumor cells reacted positively with antibodies against K8, K14, K17 and K19, but did not react, or were rarely positive with K1, K7, K10, K16 and K18 antibodies. Predominant expression of K17 and the frequent expression of K8 and K19, with little K6/K16 and K1/K10 expression are the characteristic features of BCC, suggesting that BCC is differentiated towards undifferentiated follicular epithelia, most probably hair bulge cells.

Aged↗
PubMed1998

Synchronization of normal human keratinocyte in culture: its application to the analysis of 1,25-dihydroxyvitamin D3 effects on cell cycle.

In this report, we applied the isoleucine-deprivation method to normal human keratinocytes in cultivation with serum-free MCDB153 medium. The growing cells were synchronized by transfer to MCDB153 medium prepared without isoleucine, and the degree of synchronization was analyzed by the cell cycle distribution and phosphorylation of retinoblastoma protein (pRB), one of the tumor suppressor gene products. 1,25-dihidroxyvitamin D3 (1,25(OH)2D3), a biologically active form of vitamin D3, is supposed to induce the differentiation and growth inhibition of human keratinocytes caused by cell cycle arrest. We examined its effect on cell cycle kinetics, especially progression from G1/G0 to S-phase, by using this synchronized system. We showed that 10(-6) M 1,25(OH)2D3 inhibited the progression from G1/G0 to S-phase strikingly in synchronized normal human keratinocytes.

Blotting, Northern↗
PubMed1998

Candida is not involved in the development of of periungual psoriatic lesion.

Infection can be a trigger or an aggravating factor in psoriasis, and in particular, bacterial focal infection is well documented to exacerbate psoriasis presumably through its superantigen. Since fungi could produce superantigen as well, and periungual involvement is often seen in psoriatic patients, we examined if local Candida infection plays some pathogenetic role or not in periungual lesions. In 15 psoriatic patients with periungual involvement, Candida infection was examined by microscopic study, culture, and antifungal treatment. We found no evidence suggestive of the relationship between Candida and psoriatic changes in periungual area.

Adult↗
PubMed1998

Taurin-conjugated ursodeoxycholic acid has a reversible inhibitory effect on human keratinocyte growth.

Tauroursodeoxycholic acid (TUDC) is one of the most hydrophilic taurin conjugated bile acids. TUDC has a suppressive effect on DNA synthesis in primary cultured rat hepatocytes. In this study, we investigated the growth inhibitory effect of TUDC on cultured human keratinocytes. TUDC suppressed the proliferation of keratinocytes in a dose dependent fashion, as measured by both cell counts and 5-bromo-2'-deoxyuridine (BrdU) uptake. Keratinocytes reproliferated and reached almost the same cell number as control after removal of TUDC from the medium. TUDC (1 mM) had no effect on the cell viability, as measured by the dye exclusion test. Epidermal sheets stratified in the presence of TUDC appeared thinner than those stratified without TUDC. These results suggest that TUDC has a reversible growth suppressive effect on human keratinocytes through the mechanism other than cytotoxicity and would be applicable for the treatment of hyperproliferative skin disorders such as psoriasis.

Animals↗