Biomedical subjects

Y Hoshikawa

Publications and source records attributed to Y Hoshikawa.

At least 37 records · Page 2Linked to original sources

PubMed1998

Highly controlled heterologous gene expression through combined utilization of the tetracycline-repressible transactivator and the lac repressor.

To achieve strictly on-off switching of a desired cDNA expression in a broader range of mammalian cell types, we introduced the Escherichia coli lac operator sequences into the tetracycline-repressible promoter and created a chimeric promoter (designated here as the TcIP promoter) whose activity was reciprocally controlled by tetracycline and isopropyl thiogalactopyranoside (IPTG). cDNAs were connected downstream of the TcIP promoter and stably transfected into interleukin (IL-2 or IL-3)-dependent lymphoid cells that ectopically coexpress the tetracycline-repressible transactivator and the lac repressor. Whereas the parental tetracycline-repressible promoter exhibited constitutive activities when stably introduced into the lymphoid cells, cDNA expression from the TcIP promoter was strongly inhibited by tetracycline and was potently induced by IPTG in stable transfectants. Hence, the TcIP promoter made it possible to achieve highly controlled cDNA expression in cells wherein the parental promoter did not function in an inducible manner. Potential application of this promoter was provided by expressing cyclin-dependent kinase inhibitor, p27(Kip1). Induced expression of p27(Kip1) by the TcIP promoter in the lymphoid cells strongly reduced cellular responsiveness to IL-2 or IL-3, consistent with the idea that p27(Kip1) is a critical target that must be inactivated by the interleukin-triggered mitogenic signals.

Animals↗

PubMed1998

Productive infection of Epstein-Barr virus (EBV) in EBV-genome-positive epithelial cell lines (GT38 and GT39) derived from gastric tissues.

We characterized the expression of Epstein-Barr virus (EBV) on two epithelial cell lines, GT38 and GT39, derived from human gastric tissues. The EBV nuclear antigen (EBNA) was detected in all cells of both cell lines. The EBV immediate-early BZLF 1 protein (ZEBRA), the early antigen diffuse component (EA-D), and one of the EBV envelope proteins (gp350/220) were expressed spontaneously in small proportions in the cells. EBNA 1, EBNA2, latent membrane protein 1, ZEBRA, and EA-D molecules were then observed by Western blotting in the cells. The lytic cycle was enhanced with treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) or n-butyrate. The virus particles were observed in the TPA treated GT38 cells by electron microscopy. Infectious EBV was detected with the transformation of cord blood lymphocytes and also with the induction of early antigen to Raji cells by the supernatants of both cells lines. A major single and minor multiple fused terminal fragments and a ladder of smaller fragments of the EBV genome were detected with a Xhol probe in both cell lines. These epithelial cells lines and viruses will be useful in studying their association with EBV in gastric epithelial cells.

Animals↗

PubMed1998

Control of retinoblastoma protein-independent hematopoietic cell cycle by the pRB-related p130.

The retinoblastoma tumor suppressor protein (pRB) is a potent inhibitor of mammalian cell growth and the functional inactivation of pRB is widely presumed to be essential for progression of the cell cycle from G1 phase. In this work, the generality of pRB-based cell cycle control in mammalian cells was addressed by conditionally expressing pRB in cytokine-dependent hematopoietic cells. We show herein that these cells are able to progress through the cell cycle in response to cytokine despite the continued presence of supraphysiological amounts of wild-type pRB or phosphorylation-resistant pRB mutants. However, their growth was strongly blocked by ectopic expression of the pRB-related pocket protein, p130. This growth inhibition required the E2F-binding pocket domain but not the cyclin-binding domain of p130. Furthermore, increased amounts of the p130-controlled E2F, termed E2F-4, potentiated the mitogenic response of the cells to cytokine and the constitutive overexpression of E2F-4 rendered the cells cytokine-independent. Our results indicate the existence of a non-pRB-based cell cycle whose operation depends primarily on the interplay between p130 and E2F-4 in certain hematopoietic cells.

Animals↗

PubMed1998

K-252a-induced polyploidization and differentiation of a human megakaryocytic cell line, Meg-J: transient elevation and subsequent suppression of cyclin B1 and cdc2 expression in the process of polyploidization.

Megakaryocytes are unique haemopoietic cells which undergo DNA replication, giving rise to polyploid cells. However, little is known about the mechanism of megakaryocytic polyploidization. To address this issue, we used the human megakaryocytic cell line Meg-J. In the presence of K-252a (an indolocarbasole derivative), Meg-J cells stopped proliferation and exhibited additional megakaryocytic features, including morphological changes, polyploidization, and increases in the levels of surface expression of platelet glycoprotein (GP) IIb/IIa and GPIb. Thrombopoietin (TPO) promoted the K-2 52a-induced polyploidization and megakaryocytic differentiation. In the process of K-252a-induced polyploidization, levels of expression of both cdc2 and cyclin B1 were elevated transiently and subsequently decreased. This suggested that the polyploidization process in Meg-J cells was at least in part associated with a transient elevation and subsequent decrease in the expression of cdc2/cyclin B1 complex, a critical kinase involved in G2/M cell cycle transition.

CDC2 Protein Kinase↗

PubMed1998

Pretreatment with restriction enzyme or bovine serum albumin for effective PCR amplification of Epstein-Barr virus DNA in DNA extracted from paraffin-embedded gastric carcinoma tissue.

An association between Epstein-Barr virus (EBV) and gastric carcinoma has been studied through the EBV genome present in the carcinoma cells. Recently, we found that EBV DNA in paraffin-embedded gastric carcinoma tissue was detected effectively by PCR after pretreatment of the extracted DNA with a restriction enzyme, BamHI or EcoRI. Here, we show that the PCR amplification was also enhanced by pretreatment of the DNA with other restriction enzymes or with bovine serum albumin and several other proteins. Treatment with these proteins may remove a PCR inhibitor(s) in the DNA samples extracted from the paraffin blocks.

Animals↗

PubMed1998

[Increase in pulmonary vascular permeability caused by increased adhesiveness of polymorphonuclear leukocytes and superoxide].

We report that mechanical stimulation of human neutrophils results in their accumulation in isolated rat lungs and in an increase in pulmonary vascular permeability. To determine whether reactive oxygen species were involved in this increase and, if so, whether it is mediate by xanthine oxidase metabolites, we assessed the effect of stimulated and unstimulated neutrophils, and of a superoxide scavenger, superoxide dismutase (SOD), and a xanthine oxidase inhibitor, allopurinol (ALLO) on pulmonary vascular permeability in isolated perfused lungs from Sprague-Dawley rats. Pulmonary vascular permeability in isolated rat lungs was assessed using a filtration coefficient determined by gravimetry. To quantify neutrophil accumulation in the lung, we measured myeloperoxydase (MPO). Neutrophils were stimulated by gentle agitation in a glass container for 10 s and Mac-1 was subsequently upregulated on the surface of the neutrophils. In lungs that received stimulated neutrophils, the pulmonary vascular filtration coefficient was about 5 times higher than in lungs that received unstimulated neutrophils. An increase in filtration coefficient was almost completely blocked by pretreatment with SOD or ALLO. However, the accumulation of stimulated neutrophils was not, or only partly, blocked by SOD or ALLO, respectively. We conclude that the increase in pulmonary vascular permeability caused by mechanically stimulated neutrophils was partly mediated by reactive oxygen species generated via the xanthine oxidase system.

Animals↗

PubMed1998

[Factors associated with participation in chest X-ray screening by young and middle aged residents].

We investigated the factors that affect participation rates in chest X-ray mass screenings among 20-59 year-old residents in A city, Kagawa prefecture. 1) Participation rates in employees and non-employees were 78.3% and 33.9% respectively. In the employees whose firms do not provide health check-ups, participation rate was 15.9%. 2) Among participants, about 90% of the employees whose firms do not provide health check-ups and about 60% of non-employees utilized mass screenings provided by the local government. 3) Among non-participants, 53.7% of the employees whose firms do not provide health check-ups and 70% of the others (employees whose firms provide health check-ups and non-employees) were aware of the screening system. Most of the young did not know of the system. 4) Principal reasons for nonparticipation were "inconvenient" and "unnecessary because healthy" in job holders and "unnecessary because healthy" in non-job holders. 5) Participants had better health practices than non-participants. Participation rates of chest X-ray mass screenings were high in employees and low in non-employees. Local governments are expected to supplement the existing mass screening system at work-sites by providing accessibility screening. In tuberculosis prevention, nonparticipation in mass screenings is one of the main issues. The association between participation and health practice suggests that methods for linking with primary prevention activities are necessary for stimulating interest and promoting participation in mass screenings.

Adult↗

PubMed1997

[Suppressive effect of clarithromycin on the production of verotoxin by E. coli O157].

Effects of low concentration of clarithromycin (CAM) on the production of Verotoxin (VT) by Escherichia coli O157 was investigated in vitro. The production of VT1 was suppressed up to 10 hours when bacteria was incubated with 0.64 micrograms/ml (equivalent to the 1/100th of MIC) or higher concentrations. However, the production of VT1 reached to the control level after 22 hours even with 6.4 micrograms/ml of CAM. On the other hand, production of VT2 by 22 hours was partially suppressed with 0.64 micrograms/ml of CAM and completely with 6.4 micrograms/ml of CAM. When the eight clinical isolates were incubated with 0.64 or 6.4 micrograms/ml of CAM, VT1 and VT2 were suppressed in two and eight strains, respectively. In these strains, similar but less efficient suppression of the toxin production was also observed with erythromycin. In contrast to the macrorides, ampicillin did not inhibit or rather stimulated the production of VT1 and VT2 in some strains.

Ampicillin↗

PubMed1997

Effects of Stroop color-word conflict test on the autonomic nervous system responses.

The Stroop color-word test (CWT) is a mental stress test involving sensory rejection and has been used as a model of the defense reaction in humans. The present study was designed to investigate effects of CWT on resting cardiac autonomic nervous system activity evaluated by analyses of heart rate (HR) variability (HRV). Eight healthy subjects performed 21 min of CWT after 14 min of resting control followed by 14 min of recovery (RCV). Beat-to-beat R-R intervals (RRI, i.e., HRV), systolic blood pressure (SBP), and instantaneous lung volume were continuously monitored throughout these periods. Steady-state HRV data (10-12 min) for resting control and recovery (12 min) and for 1-11 min (CWT1) and 11-21 min (CWT2) of CWT were analyzed by coarse-graining spectral analysis to break down their total power into harmonic and nonharmonic (fractal) components. The harmonic component was further divided into low (0.0-0.15 Hz, LF)- and high (>0.15 Hz, HF)-frequency components. Cardiac sympathetic (SNS) and parasympathetic (PNS) nervous system activity indicators were evaluated as LF/HF and HF/TP, respectively [LF, HF, and total spectral power (TP) of HRV]. The fractal component had the spectrum of 1/f(beta), and the spectral exponent beta was calculated. RRI decreased and SBP, plasma norepinephrine, and epinephrine concentrations increased significantly (P < 0.05) in response to CWT (CWT1 and CWT2). PNS and SNS indicators did not show any significant (P > 0.05) difference between resting controls and CWT. Whereas the percentage of fractal components in total HRV remained unchanged, beta decreased significantly (P < 0.05) in response to CWT. Transfer magnitudes from SBP to RRI, calculated by cross-spectral analysis, decreased at CWT2. The transfer magnitudes from instantaneous lung volume to RRI were also decreased significantly (P < 0.05) by CWT. It was concluded that although CWT did not affect the SNS indicator of HRV, despite altered HR and vasomotor responses, tonic sympathetic nervous system influence was observed for norepinephrine and epinephrine. In addition, reflex blood pressure (CWT2) and respiratory modulation of HR (CWT1 and CWT2) decreased during CWT. Mental stress altered the fractal nature of HRV (as judged by decreased beta), but mechanism(s) responsible for this change remained unknown.

Adult↗

PubMed1997

The stability of truncated epidermal growth factor receptor mRNA is higher than that of the full-length receptor mRNA in rat hepatoma cells.

Truncated and full-length epidermal growth factor (EGF) receptors are produced in rat liver cells. The truncated EGF receptor mRNA is almost identical to the full-length EGF receptor mRNA except for the lack of a 3' region of the full-length receptor mRNA. To understand the stability of rat EGF receptor mRNAs, we analyzed the expression of EGF receptor mRNAs in the hepatoma cell line, AH66 and liver cells. Ten, 7 and 5 kb full-length and 2.7 kb truncated EGF receptor mRNAs were detected in both of them. The half-lives of the 10 and 2.7 kb EGF receptor mRNAs were determined in AH66 cells using a transcriptional inhibitor, 5,6-dichlororibofuranosylbenzimidazole. The half-lives of the 10 and 2.7 kb mRNAs were 1.2 and 11 h, respectively. These results indicated that the truncated mRNA is 4 times more stable than the full-length mRNA in rat cells. As for the stability, the role of a sequence of 3'-untranslated region of the EGF receptor mRNA was discussed.

Animals↗

PubMed1997

Cyclooxygenase metabolites possibly produced by endothelial cells mediate the lung injury caused by mechanically stimulated leukocytes.

To determine whether mechanically stimulated leukocytes increase pulmonary vascular permeability and resistance and, if so, whether cyclooxygenase metabolites mediate the increase, we assessed the effects of stimulated and unstimulated leukocytes, and of a cyclooxygenase inhibitor on pulmonary vascular permeability and resistance in isolated perfused lungs from Sprague-Dawley rats. Leukocytes were stimulated by gentle agitation in a glass container for 10 seconds. After baseline measurements were made, stimulated or unstimulated leukocytes were added to the perfusate. The effects of the cyclooxygenase inhibitor, meclofenamate, on the pulmonary vascular filtration coefficient and pulmonary vascular resistance were measured. In the rats that received stimulated leukocytes, the pulmonary vascular filtration coefficient and the vascular resistance were about 2.5 times and 3.3 times higher, respectively, than those in the rats that received unstimulated leukocytes. These increases were completely and partly blocked by meclofenamate. Histological examination indicated that meclofenamate did not prevent the adhesion of leukocytes to the pulmonary vascular endothelium. These findings suggest that mechanically stimulated leukocytes increase pulmonary vascular permeability and that cyclooxygenase metabolites produced by endothelial cells may injure the cells.

Animals↗

PubMed1997

[MR imaging of spontaneous spinal epidural hematoma].

Spontaneous spinal epidural hematoma is a rare entity. Sudden onset of nuchal or back pain is a common presentation. Clinical diagnosis is often difficult because of its non-specific symptomatology. Irreversible neurological deficits of spinal cord may result, due to delay in initiating proper treatment. We present two cases of spontaneous spinal epidural hematoma, one in the cervical and the other in the thoracic spine. MR imaging was helpful to confirm the diagnosis and for pre-surgical planning. Both patients were treated surgically and the MR findings confirmed. MR imaging accurately localized and characterized the epidural lesions. We stress that MR imaging is a valuable examination for the diagnosis of spinal epidural hematoma.

Adolescent↗

PubMed1997

[Increase in pulmonary vascular permeability caused by increased expression of Mac-1 on the surface of polymorphonuclear leukocytes].

We studied the expression of adhesion molecules on the surface of human polymorphonuclear leukocytes (PMNs). The effects of mechanical stimulation were measured with a flow cytometer and pulmonary vascular injury due to accumulation of PMNs in the lungs was assessed by a gravimetric method. The accumulation of PMNs in the lungs was studied by measuring the amount of myeloperoxidase. PMNs were stimulated by gentle agitation in a glass container for 10 s. Mac-1 (CD11b/CD18) was upregulated on the surface of PMNs that were mechanically stimulated. When unstimulated PMNs were exposed to isolated rat lungs, the filtration coefficient did not change from that under baseline conditions. However, when mechanically stimulated PMNs were exposed to isolated rat lungs, the filtration coefficient was about 5 times higher than that measured at baseline. When mechanically stimulated PMNs treated with anti-CD18 antibody were used, the increase in the filtration coefficient was completely blocked. The assay of myeloperoxidase revealed that PMNs stuck to isolated rat lungs only after stimulated PMNs were added. We conclude that when the adhesiveness of PMNs is increased by mechanical stimulation, these cells adhere to pulmonary vessels and increase pulmonary vascular permeability.

Animals↗

PubMed1997

[Prediction of postoperative pulmonary hemodynamics for the second lobectomy after the contralateral lobectomy].

We presented 7 cases who were performed the second lobectomy for the second lung cancer after the first successful lobectomy on the contralateral lung (3 cases for right upper lobectomy + left lower lobectomy and 4 cases for right upper lobectomy + left lower lobectomy). In 6 patients, the predicted postoperative FEV1 estimated by multiplying the preoperative FEV1 by the fraction of perfusion to the contralateral lung was less than 800 ml/m2BSA, which is our first cut-off for identifying lung resection candidates. Unilateral pulmonary arterial occlusion test (UPAO) revealed that total pulmonary vascular resistance (TPVRI) in 3 of those 6 patients was lower than 700 dyne.sec.cm-5/m2BSA, our second cut-off for lung resection. More precise postlobectomy pulmonary hemodynamics in another 3 of those 6 patients were then estimated by adapting selective pulmonary occlusion test (SPAO). Since TPVRI during SPAO was lower than the cut-off value, it was suggested that second lobectomy would be feasible with low incidence of post operative cardiopulmonary complication. There was no serious complications in all 7 cases during their postoperative course. We believe that more precise prediction of postoperative pulmonary hemodynamics by adapting UPAO and SPAO could be one of the tools to minimumize postoperative cardiopulmonary complications in those patients needed second lobectomy for the second lung cancer after the first successful lobectomy on the contralateral lung even though their impaired lung fung function.

Aged↗

PubMed1996

Effects of acute exposure to simulated altitude on heart rate variability during exercise.

It has been shown that fluctuation of human heartbeat intervals [heart rate variability (HRV)] reflects variations in cardiac autonomic nervous system activity. The present study was designed to investigate whether the acute exposure to moderate levels of simulated altitude and the resultant hypoxia could modify HRV during exercise. Seven healthy men completed one resting measurement in the upright sitting position and two submaximal steady-state cycle ergometer exercises at intensities equivalent to 25 and 50% of their estimated maximal work rate. Experiments were conducted in random order at altitude equivalents of 500, 1,500, 2,500, and 3,500 m within 2 h of exposure to that altitude. Beat-to-beat HRV was measured continuously during the tests. HRV data were analyzed by "coarse-graining spectral analysis" (Y. Yamamoto and R.L. Hughson, Physica 68D: 250-264, 1993) to break down their total power (PT) into harmonic and nonharmonic (fractal) components. The harmonic component was further divided into low (0.0- to 0.15-Hz; PL)- and high (> 0.15-Hz; PH)-frequency components, and the indicators of relative sympathetic (SNS) and parasympathetic (PNS) nervous system activities were calculated by PL/PH and PH/PT, respectively. The fractal component was used to calculate the spectral exponent (beta) to evaluate the overall "irregularity" of HRV. The effects of exercise intensity (increase in heart rate, SNS indicator, and beta and decrease in PNS indicator) were significant (P < 0.05) at all altitude levels. The altitude effects (increase in heart rate and SNS indicator and decrease in PNS indicator) were found only during exercise at 3,500 m (P < 0.05). There was no significant effect of altitude on beta (P > 0.05). These data indicate that acute effects of altitude exposure on HRV were found 1) during exercise at moderate altitude (> 2,500 m) and 2) mainly for the harmonic components of HRV.

Adult↗

PubMed1996

[Effects of subacute hypoxia on alveolar epithelial ion transport in rats].

To study the effects of subacute hypoxia on alveolar epithelial ion transport, alveolar fluid clearance was measured in isolated fluid-filled rat lungs. After instillation of a solution containing about 5% bovine albumin, an increase in alveolar fluid clearance was measured over 2 hours. Alveolar fluid clearance was lower when rats were kept in hypoxic conditions (FiO2 = 0.1) for 48 hours. Neither 10(-5) M amiloride (a Na(+)-channel blocker) nor 10(-3) M ouabain (an inhibitor of Na(+)-K(+)-ATPase) inhibited fluid clearance in the hypoxia group, but they did in the normoxia group. These results indicate that subacute hypoxia may down-regulate both the Na(+)-channl and Na(+)-K(+)-ATPase, and thus decrease the absorption of excess alveolar fluid.

Animals↗

PubMed1996

[A study of surgical treatment in lung cancer over 75 years of age-effect of epidural anesthesia for protection of postoperative coughing disturbance].

29 patients over 75 years of age in 221 patients undergoing resection of lung cancer from January of 1990 through December of 1994 were studied for the occurrence of expectoration disturbance (atelectasis), the effect of epidural anesthesia for protection of it. Atelectasis was observed in 8 (27.6%) of 29. In a group received epidural anesthesia (EA) during the early postoperative phase, 1 of 5 patients developed atelectasis. In 192 patients below 75 years of age, atelectasis was observed in 40 (20.8%) of them, so in a group received epidural anesthesia (EA) during the early postoperative phase, only 2 of 42 patients (4.8%) developed atelectasis. This value was significantly lower than that value (25.3%) in a group without EA. In conclusion, in the group below 75 years of age, EA during the early postoperative phase may be useful in inhibiting the occurrence of atelectasis, an important one of the postoperative lung complications.

Aged↗

PubMed1995

[Effects of prostaglandin E1 (PGE1) on pulmonary hypertension and lung vascular remodeling in a rat monocrotaline model of human pulmonary hypertension].

Prostaglandin E1 (PGE1) has been shown to be a potent pulmonary vasodilator in humans and in many animals. The effects of PGE1 on the development of pulmonary hypertension and on pulmonary vascular remodeling were studied in a rat monocrotaline (MCT) model of human pulmonary hypertension. By 3 weeks after injection, MCT (80 mg/kg S.C.) had resulted in high values of mean pulmonary arterial pressure and of the ratio of right ventricular weight to left ventricle+septum weight (RV/LV+S). PGE1 inhibited the development of pulmonary hypertension (300 micrograms/kg) and right ventricular hypertrophy (300 and 100 micrograms/kg) induced by MCT. Three weeks after the injection, the media walls of pulmonary arteries in lungs from rats given MCT were significantly thicker than those from lungs of control rats. PGE1 (300, 100, and 30 micrograms/kg) resulted in significantly less of this morphologic change, in a dose-dependent manner. These results indicate that PGE1 inhibits the development of pulmonary hypertension associated with lung vascular thickening induced by MCT. PGE1 may be useful for the treatment of pulmonary hypertension in humans.

Alprostadil↗