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Biomedical subjects

T Ohno

Publications and source records attributed to T Ohno.

At least 343 records · Page 19Linked to original sources

Induction of human autologous cytotoxic T lymphocytes against minced tissues of glioblastoma multiforme.

The authors induced autologous cytotoxic T lymphocytes (CTLs) directly from peripheral blood lymphocytes by preparing a coculture of minced tissue fragments of glioblastoma multiforme (GBM) with interleukins-1, -2, -4, and -6 and interferon-gamma in RHAM alpha medium containing 5% autologous plasma for 2 weeks. At the end of this period, the frequencies of CD3+, CD4+, CD8+, and CD16+ lymphocytes were 95% to 99%, 40% to 62%, 37% to 38%, and 0.2%, respectively. The lymphocytes killed 82% to 100% of the GBM cells within 48 hours at an effector-to-target cell ratio of 1.67, whereas in a separate coculture, autologous lymphokine-activated killer (LAK) cells killed only 33% of GBM cells under the same conditions. The lymphocytes showed no cytotoxicity against LAK-sensitive Daudi cells, natural killer-sensitive K562 cells or autologous fibroblasts grown from the brain tumor, although they did show slight cytotoxicities against allogeneic GBM cell lines. These results lead the authors to suggest that the lymphocyte population contains specific CTLs for autologous brain tumor cells and that these CTLs could be effective in adoptive immunotherapy to combat brain tumor.

Adult↗

[Simultaneous aortic root and total arch graft replacement in a patient with aortitis syndrome].

The patient was a 45-year-old female diagnosed with aortitis syndrome and aortic regurgitation (AR). She had been taking steroid therapy since 1975. She had recently developed congestive heart failure due to AR while both the ascending aorta and aortic arch were enlarged. She had no inflammatory reaction on admission. An aortogram showed heavy dilation of both the ascending aorta and aortic arch and maximum diameters was 11 cm in the ascending aorta and 4.5 cm in the descending aorta. There was an obstruction of the left subclavian artery. Moderate AR was seen on an echocardiogram. She had a simultaneous graft replacement of aortic root and total arch. The aortic root was replaced with composite graft and coronary arteries were implanted using Carrel's patch technique, and the aortic arch was also replaced with a graft with two side branches. The postoperative course was uneventful without complication of cerebral infarction or paraplegia. The postoperative aortogram showed stenosis of the left carotid artery, but no abnormality of the coronary orifices and graft anastomosis. She returned home with disappearance of symptoms of congestive heart failure.

Aorta↗

Development of a centrifugal pump with improved antithrombogenicity and hemolytic property for chronic circulatory support.

A centrifugal pump with a unique structure has been developed for chronic support. The pump is driven by a magnetic coupling and has no rotating shaft, no seal around the rotating part, and a balancing hole at the center of the impeller and the thrust bearing. The pump was improved in stepwise fashion to realize good antithrombogenicity and low hemolysis. The first pump, the National Cardiovascular Center (NCVC)-0, had an impeller with 4 rectangular and curved vanes; 6 triangularly shaped curved vanes were employed in the second model, the NCVC-1, to reduce trauma to the blood. In the third design, the NCVC-2, the central hole was enlarged, and the thrust bearing shoulder was rounded so that blood washing was enhanced around the impeller; stream lines also were smoothed for improved antithrombogenicity. The hemolytic property of the device was evaluated in vitro with heparinized fresh goat blood; hemolysis indexes of the NCVC-0, -1, and -2 were 0.05, 0.01, and 0.006 g per 100 L, respectively. Antithrombogenicity of the pumps was examined in animal experiments as a left heart bypass device in goals weighing 52-75 kg. Six NCVC-0 pumps were driven for 14 to 33 (22.0 +/- 7.6) days in goats receiving the antiplatelet drug cilostazol orally. Four NCVC-1 pumps ran for 1 to 80 (28.5 +/- 30.6) days with the same drug regimen in 2 cases and with no anticoagulation therapy in 2 cases. After 3 preliminary 1-week tests of NCVC-2 pumps in animals, the pump was installed in 3 goats; 2 pumps were still running on the 182nd and 58th pumping day. Intracorporeal implantation also was attempted successfully. The results indicate that this pump has promising features for chronic support although longer term and additional evaluations are necessary.

Animals↗

[Temperature changes and thermoregulatory responses during epidural anesthesia in women undergoing cesarean delivery].

Eleven healthy pregnant women were studied to determine temperature changes after induction of epidural anesthesia during cesarean delivery. Epidural anesthesia was induced by 2% lidocaine 15-20 ml (T 5 level) at 25 degrees C ambient temperature. Tympanic membrane and skin surface temperature, skin-temperature gradients (forearm-fingerchip, calf-toe), thermal perception (1-10 scale VAS) and the presence or absence of shivering were measured. Sixty minutes after induction, tympanic temperature decreased for 0.52 +/- 0.26 degrees C and average skin temperature increased for 0.56 +/- 0.17 degrees C. Central hypothermia and shivering did not produce a cold sensation. Temperature gradients of upper limb increased to 2.0 +/- 3.7 degrees C (50 minutes after induction) and the lower limbs decreased to -1.4 +/- 0.55 degrees C (30 minutes after induction). Shivering occurred in 3 patients. We conclude that epidural anesthesia impaired thermoregulatory control and induced redistribution hypothermia as in nonpregnant individuals.

Adult↗

[Myelodysplastic syndrome with CREST syndrome successfully treated with metenolone--A case report].

A 54-year-old woman was diagnosed as having refractory anemia (RA) with CREST syndrome (incomplete type). She showed Raynaud's phenomenon, sclerodactyly and telangiectasia, but not calcinosis and esophageal dysmotility. Laboratory findings revealed anemia and thrombocytopenia, and myelodysplasia, abnormal karyotype of 47, XX, +8 in bone marrow cells. Antinuclear and centromere antibody was positive. Treatment with prednisolone was not successful. After prednisolone was tapered, she was given 20 mg/body metenolone orally, which led to hematological improvement, and after 6 months of therapy, abnormal karyotype of 47, XX, +8 disappeared.

Anabolic Agents↗

[An analysis of slow phase velocity on congenital nystagmus-asymmetrical responses of optokinetic stimulation].

The responses of optokinetic nystagmus (OKN) were examined in 11 patients with congenital nystagmus (CN). When the nystagmus was almost suppressed in a dark room, the OKN stimulation was started. There were three different type of OKN responses: bilaterally reversed OKN, unilaterally, reversed OKN and pendular oscillation. Three of 11 patients had normal direction in the slow phase of OKN in one direction, but reversed OKN in the other direction. These findings suggest that two different directional slow eye movement subsystems (rightward and leftward) exist in CN patients and that the mechanism of CN is unilateral or bilateral dysfunction of the two different directional slow eye movement subsystems.

Adolescent↗

[Intraventricular arachnoid cyst--on the origin of intraventricular arachnoid cysts].

Arachnoid cysts very rarely occur within the ventricular system, where no arachnoid tissue exists. We present three cases of intraventricular arachnoid cyst with special reference to its origin. The first patient was a 5-year-old boy who complained of headaches and enlargement of his head. A CT scan revealed obstructive hydrocephalus and a large cystic lesion in the right lateral ventricle. The symptoms resolved after fenestration of the cyst and cystoperitoneal shunt. The second patient was a 49-year-old woman who complained of headache and numbness in her left upper extremity. A CT scan and MRI revealed a large cyst in the trigone of the right lateral ventricle. Fenestration of the cyst wall and cystoperitoneal shunt were performed relieving her complaints. The third patient was a 42-year-old man who complained of frequent seizures and dizziness. A CT scan and MRI demonstrated a moderate size cystic mass in the inferior horn of the right lateral ventricle. The symptoms were improved by partial resection of the cyst wall. Immunohistochemical studies and light microscopy confirmed that the cyst walls were composed of arachnoid membrane, implying that the cysts were arachnoid cysts. CT and MRI in these three cases showed widening of the choroidal fissure bordering the cyst wall, occasionally involving a part of the protruding cyst wall. On enhanced CT and MRI, the choroid plexus in the trigone of ipsilateral lateral ventricle was displaced anterolaterally, implying that the cysts had grown from outside the choroid plexus. Postoperative MRI demonstrated the shrunken cyst wall attached to the choroidal fissure. The surgical findings in case 3 also showed that the cyst wall was attached firmly to the choroid plexus. These findings appeared to indicate that the intraventricular arachnoid cyst originated from the arachnoid layer drawn into the choroidal fissure with choroidal vascular mesenchyme.

Adult↗

Frequent expression of P-glycoprotein/MDR1 by nasal T-cell lymphoma cells.

BACKGROUND: Lethal midline granuloma is now considered to be a malignant lymphoma derived from peripheral T cells or from natural killer cells. The therapeutic outcome of nasal T-cell lymphoma (NL) treated by conventional chemotherapy for non-Hodgkin's lymphoma is poor, although some patients have a good response to radiotherapy. To clarify the mechanisms of drug resistance, the expression of P-glycoprotein (P-gp)/MDR1, which is the product of the multidrug resistance (MDR) 1 gene, and MDR3 mRNA in NL cells, were examined. METHODS: Ten Japanese patients with NL were studied. Nine of these patients were examined before therapy. P-glycoprotein expression and phenotypes of lymphoma cells were examined by immunohistochemical staining using UIC2 as an anti-P-gp monoclonal antibody. In one case, the Rhodamine-123 efflux test was performed. MDR1 and MDR3 mRNA were detected by reverse transcription polymerase chain reaction. RESULTS: Nine of the 10 patients were P-gp positive. In one of nine, functional P-gp expression was observed. MDR1 mRNA was detected in all seven examined patients with P-gp positive NLs, whereas MDR3 mRNA was negative. Retrospectively, patients who received chemotherapy alone had poorer outcome than those treated by combination chemotherapy after irradiation. CONCLUSION: The poor prognosis for patients with NL treated with chemotherapy may be explained by P-gp expression of the NL cells.

ATP Binding Cassette Transporter, Subfamily B↗

Detection of arginine-ADP-ribosylated protein using recombinant ADP-ribosylarginine hydrolase.

We made use of ADP-ribosylarginine hydrolase to detect arginine-ADP- ribosylated proteins. The hydrolase was expressed in Escherichia coli as a protein fused with glutathione S-transferase (GST). The fusion protein GST-ADP-ribosylarginine hydrolase catalyzed the hydrolysis of alpha-ADP-ribosylarginine to produce ADP-ribose and arginine. Casein ADP-ribosylated with [32P]NAD and chicken heterophil arginine-specific ADP-ribosyltransferase served as a substrate for the recombinant ADP-ribosylarginine hydrolase and the released ADP-ribose was determined. Protein ADP-ribosylated by cholera toxin could serve as substrate of the hydrolase but protein ADP-ribosylated by pertussis toxin, diphtheria toxin, or C(3) enzyme of Clostridium botulinum could not. The hydrolase did not release the radioactivity incorporated into isolated rat liver nuclei incubated with [(32)P]NAD or in bovine brain cytosol incubated with [(32)P]ADP-ribose. In homogenate of mouse heart which contained arginine-specific ADP-ribosyltransferase, labeling of a 55-kDa protein by incubation with [(32)P]NAD was removed by ADP-ribosylarginine hydrolase treatment; hence, the specific hydrolysis of ADP-ribose-arginine bond by GST-ADP-ribosylarginine hydrolase can be used to detect the arginine-ADP-ribosylated proteins in crude preparations. Arginine--ADP-ribosylated proteins in crude preparations. Arginine-ADP-ribosylated proteins in mouse spleen lymphocytes were identified using this method.

Adenosine Diphosphate Ribose↗

Loss of tumorigenicity of Ewing's sarcoma cells expressing antisense RNA to EWS-fusion transcripts.

Cytogenetic analysis of Ewing's sarcoma, primitive neuroectodermal tumors and Askin tumors revealed characteristic translocations t(11;22) or t(21;22). Molecular analysis of these translocations revealed 5'-region of EWS gene (from band 22q12) is fused to the 3'-region of either Fli-1 gene (from band 11q24) or erg gene (from band 21q22). Functional characterization of the EWS-Fli-1 and EWS-erg chimeric proteins suggested that they function as transcriptional activators. In order to develop therapeutic agents, it is essential to know whether expression of the EWS-fusion gene products is coupled to tumorigenicity of Ewing's sarcoma cells and if targeting the EWS-fusion products results in loss of tumorigenicity of Ewing's sarcoma cells. For this reason, we have made stable Ewing's sarcomas expressing antisense EWS-Fli-1 or EWS-erg expression plasmids. Expression of antisense EWS fusion transcripts resulted in a significant loss of endogenous EWS-Fli-1 and EWS-erg proteins in Ewing's sarcoma cells. These cells expressing antisense EWS fusion transcripts showed loss of anchorage independent growth and tumorigenicity in nude mice unlike the parental Ewing's sarcoma cells. These results demonstrate the necessity of a certain threshold level of expression of EWS-fusion products in the clonogenicity and tumorigenicity of Ewing's sarcoma cells and therefore emphasizes the importance of targeting the EWS-fusion products as a therapy for Ewing family of tumors.

Animals↗

Myosin-actin interaction plays an important role in human immunodeficiency virus type 1 release from host cells.

We examined the potential role of myosin and actin in the release of human immunodeficiency virus type 1 (HIV-1) from infected cells. Wortmannin (100 nM to 5 microM), an effective inhibitor of myosin light chain kinase, blocked the release of HIV-1 from infected T-lymphoblastoid and monocytoid cells in a concentration-dependent manner. Cytochalasin D, a reagent that disrupts the equilibrium between monomeric and polymeric actin, also partially inhibited the release of HIV-1 from the infected cells. At the budding stage, myosin and HIV-1 protein were detected in the same areas on the plasma membrane by using dual-label immunofluorescence microscopy and immunoelectron microscopy. In the presence of 5 microM wortmannin, viral components were observed on the plasma membrane by using immunofluorescence microscopy and electron microscopy, implying that wortmannin did not disturb the transport of viral proteins to the plasma membrane but rather inhibited budding.

Actins↗

Regulation of human retroviral latency by the NF-kappa B/I kappa B family: inhibition of human immunodeficiency virus replication by I kappa B through a Rev-dependent mechanism.

The cellular transcription factor NF-kappa B stimulates human immunodeficiency virus type 1 (HIV-1) transcriptional initiation, but its role in the retroviral life cycle has not been fully defined. In this report, we show that I kappa B alpha acts as a cellular inhibitor of human retroviral replication through a discrete mechanism, independent of its effect on HIV transcription. I kappa B alpha inhibited HIV replication and gp160 expression by negatively regulating Rev function, most likely acting through a cellular factor involved in Rev transactivation. A similar effect was observed with human T leukemia virus I, in which I kappa B alpha inhibited Rex function. In contrast, no effect was observed on the replication of a DNA virus, adenovirus type 5. The NF-kappa B/I kappa B regulatory pathway therefore modulates human retroviral replication by regulating a program of cellular gene expression required for several steps in the viral life cycle, including not only viral transcription but also RNA export. This interaction between cellular and viral gene products suggests that NF-kappa B plays a broader role in the regulation of human retroviral replication, providing a previously unrecognized link between two important regulators of HIV gene expression and common NF-kappa B-dependent programs of gene expression used by human retroviruses.

Base Sequence↗

C-type natriuretic peptide inhibits intimal thickening after vascular injury.

Recently we have found that C-type natriuretic peptide (CNP) inhibits proliferation of cultured rat vascular smooth muscle cells through an elevation of cGMP. We have now tested whether administration of CNP inhibits the development of intimal lesions induced by air-drying injury in rat common carotid arteries in vivo. CNP treatment (1 microgram/kg per min, iv infusion) for either 14 or 5 days resulted in 70% or 60% reduction, respectively, of intimal cross-section area 14 days after injury as compared with control rats. We also found that CNP potently stimulated cGMP production in injured carotid arteries with intimal thickening, but not in intact ones. These results indicate that GC-B, CNP specific receptor/guanylyl cyclase, is expressed at the sites of vascular injury, and that CNP might be efficacious in the prevention of restenosis caused by intimal thickening following coronary angioplasty.

Animals↗

Measurement of yeast intracellular pH by image processing and the change it undergoes during growth phase.

The intracellular pH of the yeast Saccharomyces cerevisiae was determined by a fluorescence microscopic image processing technique. Image processing was carried out using a modification of the ratio imaging method for measurement of yeast intracellular pH. Care was necessary when taking fluorescence images in order to obtain accurate measurement of yeast intracellular pH. Until now it has been difficult to measure the intracellular pH of cells in actual cultivation conditions. This method enabled us not only to measure the intracellular pH of dilute cell suspensions, but also to obtain two-dimensional information. In the case of resting cells, the intracellular pH was dependent upon the extracellular pH, and this value was constant when the extracellular pH was constant. On the other hand, in the case of actively growing cells, intracellular pH was found to change, even if the extracellular pH was constant: the values observed were intracellular pH 5.7 during lag phase, intracellular pH 6.8 during exponential phase and intracellular pH 5.5 during stationary phase. These results for intracellular pH indicate that the yeast proton pump was activated during growth from the point of view of pH in vivo.

Ammonia↗

Seroprevalence of hepatitis C virus infection and its genotype in Lanzhou, western China.

The seroprevalence of hepatitis C virus (HCV) infection in Lanzhou, Western China was studied. HCV genotypes in 20 patients with HCV infection was determined by genotype-specific primer for polymerase chain reaction (PCR) based on HCV core region and compared with the genotype assigned by sequence comparison and molecular evolutionary analysis based on the same region. Antibody to HCV (anti-HCV) was present in 2.5% of volunteer blood donors and in 35.0% of paid blood donors (P < 0.01). HCV infection is uncommon in patients with liver disease who attended liver clinics in this locality; 4.0% with acute hepatitis and 4.0% with chronic hepatitis, 10.0% with liver cirrhosis, and none with hepatocellular carcinoma were seropositive for anti-HCV. Genotype 1b and 2a were both found to be prevalent. Together, they accounted for 19 of 20 (95%) patients with HCV infection. Sequencing of the HCV core region from two patients showed that the assignment of HCV genotype by genotype-specific primers for PCR matched well with the genotyping results based on sequence comparison and molecular evolutionary analysis. These data showed that HCV is present in Western China, HCV infection is more common in paid blood donors, and HCV genotypes 1b and 2a are both prevalent in Western China.

Amino Acid Sequence↗

Loss of serum HCV RNA at week 4 of interferon-alpha therapy is associated with more favorable long-term response in patients with chronic hepatitis C.

To determine the virological factors associated with a favorable long-term response to interferon-alpha (IFN) therapy in chronic hepatitis C virus (HCV) infection, 61 Japanese patients with chronic HCV infection were treated with IFN for 24 weeks (780 million units in total) and followed for 8 to 16 months after cessation of therapy. Ten patients dropped out because of severe side effects. Of the 51 patients who completed IFN therapy, 23 showed complete and sustained response (CR --> SR), 13 complete response with early relapse (CR --> Rel), and 15 no response to IFN (NR). For the pretreatment serum HCV RNA level, 20/23 who had CR --> SR had < 10(6) eq/ml compared to 3/13 CR --> Rel and 1/15 NR (P < 0.01). Serologically defined HCV type 2 infection was also associated with a better opportunity to develop CR --> SR compared to CR --> Rel of NR (P < 0.01). Loss of serum HCV RNA at week 4 of IFN therapy was also associated with a more favorable long-term response [17/19 CR --> SR were HCV RNA negative compared to 3/11 CR --> Rel (P < 0.01) and 2/13 NR (P < 0.01)]. In contrast, normalization of serum alanine aminotransferase (ALT) levels at week 4 was found in 9/19 CR --> SR compared to 8/11 CR --> Rel (P = NS), and 0/13 in NR (P < 0.01). Six months after cessation of IFN therapy, 3/25 CR --> SR patients were HCV RNA positive despite normalization of serum ALT levels. These data indicated that in addition to pretreatment serum HCV RNA levels and HCV type, the kinetics of response to IFN (at week 4) were also predictive of subsequent long-term response to IFN in patients with chronic HCV infection.

Adult↗