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Biomedical subjects

A Maeda

Publications and source records attributed to A Maeda.

At least 145 records · Page 8Linked to original sources

Intramuscular injection of plasmid DNA expressing mRNA7 coding the nucleocapsid protein of JHMV partially protected mice against acute infection with JHMV.

We constructed a plasmid expressing mRNA7 coding the nucleocapsid (N) protein of JHM strain of mouse hepatitis virus (JHMV) under the control of Rous sarcoma virus LTR, referred to as pRSV-mRNA7. When C57BL/6 mice injected intramuscularly (i.m.) with control plasmid DNA which contained no viral sequence were infected with JHMV, necrotic figures of neural cells and diffuse immersion of lymphatic cells in the pedunculus cerebri were observed. In the hypothalamus, vascular cuffing consisting of lymphatic cells was observed. In contrast, no histological change was observed throughout these areas of the brains in the JHMV-infected mice after injection with pRSV-mRNA7. These results showed that the injection with plasmid DNA expressing mRNA7 of JHMV partially protected mice against acute infection with JHMV in the brain. The plasmid DNA was i.m. injected into mice and the cytolytic activity of spleen cells from the mice was assessed by 51Cr-release assay. The spleen cells from the mice administrated with pRSV-mRNA7 showed a significant level of cytolytic activities against the transfected cells expressing the viral N protein even though the spleen cells were not cocultivated with stimulator cells. When the spleen cells from administrated mice with pRSV-mRNA7 were cocultivated with stimulator cells, higher cytolytic activity was observed against the transfected cells, compared with the activity without stimulation.

Acute Disease↗

Inhibitory effects of modified oligonucleotides complementary to the leader RNA on the multiplication of mouse hepatitis virus.

Phosphorothioate oligonucleotides (PS-oligo) and PS-oligos with cholesterol conjugates (ChPS-oligo) complementary to the leader RNA of strain JHM of mouse hepatitis virus (JHMV) were more effective inhibitors of viral multiplication than natural oligodeoxynucleotides (PO-oligo) in JHMV-infected DBT cells. PS- and ChPS-oligos were 1,000 times more potent than unmodified PO-oligo. No significant difference was observed in the inhibitory efficiency between PS-oligo and ChPS-oligo. Sequence-dependent inhibition of viral multiplication was shown at low concentrations (0.001-0.1 M) of antisense PS-oligo and ChPS-oligo. Phosphorothioate oligodeoxycytidine, PS-(dC)20, and PS-(dC)20 with cholesterol conjugates, and PS- and ChPS-oligo which have no significant homology to the JHMV sequences, showed inhibitory effects on JHMV multiplication at concentrations higher than 0.5 M. These results showed that PS-oligo and ChPS-oligo were more potent than PO-oligo in the inhibition of JHMV multiplication, and that PS-oligo and ChPS-oligo may inhibit JHMV multiplication by two different mechanisms, that is by sequence-dependent and sequence-independent manners.

5' Untranslated Regions↗

Effect of omeprazole and cimetidine on plasma aldosterone response to angiotensin II.

OBJECTIVE: The Effect of omeprazole, a proton pump inhibitor, and cimetidine, and H 2-receptor antagonist, on plasma aldosterone (PA) response to angiotensin II (AII) were evaluated. METHODS: Furosemide (a loop diuretic agent, 20 mg) was given intravenously to eight healthy subjects during a control period, and after pretreatment with omeprazole (20 mg daily) or cimetidine (800 mg daily) for 6 days. Blood samples for determination of plasma renin activity (PRA), AII, PA, adrenotorticotropic hormone (ACTH) and potassium were obtained just before, and 30, 60 min and 120 min after furosemide administration. RESULTS: PRA, AII and PA increased significantly after furosemide administration whereas ACTH and potassium did not. Significant correlations between plasma AII and PA were obtained in the control and omeprazole trial, but not in the cimetidine trial. The slope of the regression lines of the control and omeprazole trails did not differ significantly. CONCLUSION: These results suggest that, in contrast to cimetidine, the inhibitory effect of omeprazole on AII-stimulated aldosterone production following dosing with furosemide is negligible.

Adrenocorticotropic Hormone↗

Pharmacokinetic alterations of quinapril during repeated treatment in elderly subjects.

OBJECTIVE: To examine whether the pharmacokinetics of quinaprilat, an active metabolite, change during repeated treatment with quinapril, an ACE inhibitor, in elderly subjects. METHODS: Quinapril (10 mg) was given once daily for 8 days in eight elderly hypertensive subjects (76 years old). Blood samples were obtained for a 24-h period after the first and eighth doses. RESULTS: Plasma concentrations of quinaprilat after the eighth dose significantly higher than those after the first dose. The maximum plasma concentration (Cmax) tended to be greater, and the area under the plasma concentration-time curve (AUC) was significantly greater after the eighth dose. CONCLUSIONS: The study showed an increase in quinaprilat concentrations and subsequent increase in AUC, during repeated treatment with quinapril in elderly subjects. However, the differences observed were very small and of no clinical significance.

Aged↗

Endothelial cell integrin laminin receptor expression in multiple sclerosis lesions.

Laminin, a major glycoprotein component of vessel basement membranes, is recognized by beta1- and beta3-integrins expressed on endothelial cells. To determine how endothelial cell integrins might function in multiple sclerosis (MS) lesions, integrin laminin receptors and laminin were analyzed in central nervous system samples from MS patients and controls by immunohistochemistry. In active MS lesions, endothelial cell VLA-6 and beta1 subunits were decreased compared to controls whereas alpha(v) subunit and VLA-1 were increased. In chronic inactive lesions beta1, VLA-6 and alpha(v) were the same as controls but VLA-1 remained increased. Alpha3 subunit was constant in all samples. By immunoelectron microscopy VLA-1, VLA-6, beta1, and laminin were distributed throughout endothelial cells; alpha(v) was adjacent to and on luminal surfaces; alpha(v) and VLA-1 were on intercellular junctions. These results indicate distinct regulation and functions of these integrins in different lesion stages. In active lesions decreased endothelial cell beta1/VLA-6 could result in their detachment from laminin thereby facilitating leukocyte transvascular migration and blood-brain barrier breakdown. Alpha(v) and VLA-1 on intercellular junctions may participate in re-establishing vessel integrity after leukocyte migration. Luminal surface alpha(v) also likely binds intraluminal ligands and cells. In chronic inactive plaques persistently elevated endothelial cell VLA-1 correlates with long-standing endothelial cell and blood-brain barrier dysfunction.

Adolescent↗

Body support effect on standing balance in the visually impaired elderly.

OBJECTIVE: To evaluate the effectiveness of body support in compensating for decreased standing balance stability in elderly persons with visual impairment. DESIGN: Standing balance was measured in a cross-section of elderly persons from two nursing homes--one for aged individuals with visual impairment, the other for aged individuals without visual impairment. PARTICIPANTS: The subjects were 44 visually impaired persons with a mean age of 79.0 yrs and 39 people without visual impairment, mean age 76.3 yrs. OUTCOME MEASURES: The area of gravity-center sway was measured with subjects standing on a gravicorder for 30sec in three positions: (1) without support, (2) with cane, (3) with light support by touching a wall. RESULTS: Both men and women in the visually impaired group swayed more than their sighted counterparts when standing without support, the only statistically significant difference between the two groups. For all subjects, the greatest degree of sway occurred when subjects stood unsupported, and the least sway occurred when subjects touched a wall for support. CONCLUSION: In visually impaired elderly persons, touching a wall for body support while standing is more effective than using a cane.

Aged↗

Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription.

The (-)-strand viral RNAs that result from after infection of cells with coronaviruses, which possess RNA genomes of message polarity, are genomic-sized and subgenomic-sized. Each of the (-)-strand subgenomic RNAs corresponds in size to each of the subgenomic mRNA species that are made in infected cells. We tested whether (-)-strand subgenomic RNAs might initially be synthesized from the input single-stranded (+)-strand genomic RNA prior to the production of subgenomic mRNAs. We used a mouse hepatitis virus (MHV) defective interfering (DI) RNA. from which subgenomic RNA was produced in DI RNA-replicating cells, because this DI RNA had a functional MHV intergenic region inserted in its interior. MHV samples containing the DI particles were irradiated with UV-light and then superinfected into cells that had been infected with MHV 4 h prior to superinfection. Northern blot analysis of intracellular RNAs that were extracted 3 h after superinfection showed that genomic DI RNA and subgenomic DI RNA had similar UV-target sizes, indicating that (-)-strand genomic DI RNA synthesis from input genomic DI RNA probably occurred prior to the subgenomic-size DI RNA synthesis. We discuss why, in the course of coronavirus transcription, (-)-strand genomic-length coronavirus RNA synthesis might occur before subgenomic-sized RNAs of either polarity are made.

Animals↗

In vivo fate of folate-BSA in non-tumor- and tumor-bearing mice.

KB tumor cells exhibit an increased number of folate receptors on their membrane. This receptor has been proposed as a promising target for tumor drug targeting. Therefore, the disposition of folate-conjugated bovine serum albumin (folate-BSA) was examined as a model system for drug targeting. Nude mice which had received KB tumor cell transplants were given bolus intravenous administration of either 111In-labeled folate-BSA (111In-folate-BSA; 1 mg/kg) or unmodified 111In-BSA (111In-BSA; 1 mg/kg). The disposition characteristics and pharmacokinetics of 111In-folate-BSA were compared with those of the 111In-BSA as a control. The half-life of the beta-phase of 111ln-folate-BSA in plasma was 140 min. The tumor uptake rate index for 111In-folate-BSA was 0.46 microL/min/g, and that for 111In-BSA was 0.32 microL/min/g. This index of 111In-folate-BSA was slightly higher than that of 111In-BSA in vivo, by a factor of 1.4. In vivo experiments showed folate-BSA has a relatively long plasma duration. 111In-folate-BSA also showed selective distribution to tumors, but not as great as recent results from in vitro experiments. Therefore, the low vascular permeability of BSA into solid tumor tissue and inhibition of folate-mediated 111In-folate-BSA uptake by tumor cells from the blood may be the rate-limiting factor of distribution.

Animals↗

Associations between anthropometric indices of adiposity and atherogenic risk factors in Japanese working women aged 21-40 years.

This study was conducted to explore whether anthropometric indices of obesity are associated with atherogenic risk factors in young adult working women in Japan. The subjects were 492 women in an occupational setting. Predictor variables were body mass index (BMI), the sum of triceps and subscapular skinfold thickness (SFT), and the waist to hip ratio (WHR). Outcome variables were serum total cholesterol, triglyceride and blood pressures. The average age of the subjects was 26.3 (SD 3.9) years. The upper quartiles of BMI and SFT were significantly associated with all atherogenic risk factors, while the upper quartiles of WHR were not. Multiple comparisons revealed the 4th quartiles of BMI (> 22.25) and SFT (> 39 mm) to have significantly higher values for all atherogenic risk factors. We found that BMI and skinfold thickness were more relevant to the prediction of atherogenic risk factors than WHR in young adult Japanese women.

Adipose Tissue↗

Expression of CD95 ligand in melanocytic lesions as a diagnostic marker.

CD95 ligand (CD95L) potently induces apoptosis by activating CD95 on target cells. It has recently been reported that melanoma cells in vivo express a significant amount of CD95L, thereby being immediately able to kill CD95-bearing immunocompetent cells specific for cancer antigens, which infiltrate the lesions. In this study, we employed immunohistochemistry using an antibody directed against CD95L to investigate at which stage the melanoma CD95L expression is turned on. Skin biopsies of 49 lesions from 46 patients were assessed. These included benign and dysplastic naevi, melanoma in situ, stage I melanomas (Clark's level 2 or 3), advance-phase melanomas (Clark's level 4 or 5) and lymph node metastases. CD95L was expressed in all of the advance-phase melanomas as well as lymph node metastases of cutaneous origin, whereas neither melanoma in situ, benign naevi nor dysplastic naevi reacted positively with the antibody. To investigate a link between positivity and tumour size, the data were analysed on the basis of Breslow thickness, and indicated that expression was observed only when tumours were thicker than 0.75 mm. We next compared expression of CD95L and HMB-45. CD95L was positive only in melanomas in a more advanced phase than stage I, whereas HMB-45 was not only expressed in melanoma cells but also in benign pigmented naevi. This indicated the advantage of CD95L staining to diagnose melanoma. The present study indicates the significant correlation between tumorigenicity and expression of CD95L, and thereby raises the possibility that CD95L may be a useful diagnostic marker for malignant melanomas.

Antigens, Surface↗

The expression of co-stimulatory molecules and their relationship to the prognosis of human acute myeloid leukaemia: poor prognosis of B7-2-positive leukaemia.

We examined the expression of co-stimulatory molecules on leukaemic cells of 52 adult patients with acute myeloid leukaemia (AML) (34 men and 18 women) and analysed the relationship between these expressions and the patient's prognosis. B7-1 was not expressed in any of the 23 patients investigated, whereas B7-2 was expressed in 26/52 patients (50.0%). B7-2 was expressed in all AML patients with monocytic morphology (M4 or M5) and in 16/42 cases without monocytic morphology. CD54 was expressed in 28/ 37 patients examined (75.7%), and CD58 was expressed in all of the AML patients except one (M 7). The overall survival of the 26 B7-2-positive leukaemia patients (1-24 months, median survival 11.5 months) was significantly shorter than that of the 26 B7-2-negative leukaemia patients (1-71+ months, median 35.1 months) (P=0.0080). In addition, the B7-2-positive patients exhibited significantly shorter disease-free survival periods compared to the B7-2-negative patients (P=0.021). There was no significant difference in age, sex, haematological data and complete remission rate between the B7-2-positive and B7-2-negative patients. Our results indicated that B7-2 is one of the most crucial factors in the prognosis of adult acute leukaemia and can be expected to have an important role in tumour immunity.

Acute Disease↗

Alteration of circadian time structure of blood pressure caused by night shift schedule.

The effects of night shift schedules on circadian time structure of blood pressure were studied in seven healthy young subjects by continuous monitoring of blood pressure every 30 min for 72 h. In the control experiment, subjects were instructed to sleep at regular times with the light off at 00.00 h and the light on at 07.00 h. In the shift experiment, they were instructed to go to bed at 06.00 h and wake up at 11.00 h. The circadian rhythm of blood pressure rapidly phase delayed by 3.5 h in the second night shift day as a group phenomenon. Individual differences in changes in power spectral patterns of blood pressure were found in the night shift schedule. Ultradian rhythmicity of blood pressure was more pronounced in three subjects, whereas the circadian rhythmicity was maintained in four subjects. These findings held when the adaptation to shift work was taken into account.

Adult↗

Periodic discharge of adrenocorticotropin and vasopressin associated with focal glomerulosclerosis.

We report the first case of the syndrome of periodic adrenocorticotropin (ACTH) and vasopressin (ADH) discharge associated with focal glomerulosclerosis. Approximately 30 cases of this syndrome have so far been reported in Japan, but no cases associated with renal dysfunction have yet been reported. The patient, a 10-year-old Japanese boy, was referred to our hospital because of recurrent attacks of vomiting. He was diagnosed as having this syndrome from clinical and laboratory findings. While various drugs were tried to manage his vomiting attacks, only valproic acid appeared to be effective in reducing the frequency of the attacks. Chronic nephritis was manifested when the patient was 12 years old, which required treatment with continuous ambulatory peritoneal dialysis. Valproic acid was proved to be effective in reducing the number of attacks over 4 months.

Adrenocorticotropic Hormone↗

Application of subtype-specific monoclonal antibodies for rapid detection and identification of influenza A and B viruses.

We established a rapid method for the identification of influenza A and B virus strains: the peroxidase-antiperoxidase (PAP) staining method with two subtype-specific murine monoclonal antibodies, C179 (H1 and H2 specific) and F49 (H3 specific), and an anti-influenza B virus rabbit polyclonal serum. The types and subtypes of 160 strains were examined, and 158 strains were identified to be the same by the hemagglutination-inhibition (HI) test and the PAP method. In contrast to the results by the HI test, two strains were revealed to be a mixture of two subtypes (H1 and H3) by the PAP method, which was confirmed by plaque cloning. We further analyzed clinical specimens by the PAP method by directly inoculating specimens into Madin-Darby canine kidney cells in microplates. After 40 h of incubation, the types and subtypes of viruses in 52 of 152 specimens were clearly identified. Since the reactivities of the two monoclonal antibodies are not influenced by the antigenic drift of influenza virus, the newly developed method should be applicable not only for rapid diagnosis but also for the epidemiological study of influenza.

Animals↗

Coronavirus transcription early in infection.

We studied the accumulation kinetics of murine coronavirus mouse hepatitis virus (MHV) RNAs early in infection by using cloned MHV defective interfering (DI) RNA that contained an intergenic sequence from which subgenomic DI RNA is synthesized in MHV-infected cells. Genomic DI RNA and subgenomic DI RNA accumulated at a constant ratio from 3 to 11 h postinfection (p.i.) in the cells infected with MHV-containing DI particles. Earlier, at 1 h p.i., this ratio was not constant; only genomic DI RNA accumulated, indicating that MHV RNA replication, but not MHV RNA transcription, was active during the first hour of MHV infection. Negative-strand genomic DI RNA and negative-strand subgenomic DI RNA were first detectable at 1 and 3 h p.i., respectively, and the amounts of both RNAs increased gradually until 6 h p.i. These data showed that at 2 h p.i., subgenomic DI RNA was undergoing synthesis in the cells in which negative-strand subgenomic DI RNA was undetectable. These data, therefore, signify that negative-strand genomic DI RNA, but not negative-strand subgenomic DI RNA, was an active template for subgenomic DI RNA synthesis early in infection.

Animals↗

Evaluation of antibodies to the Epstein-Barr virus immediate early gene product ZEBRA by a new enzyme-linked immunosorbent assay.

For the serodiagnosis of Epstein-Barr virus (EBV) infections, we have developed a new enzyme-linked immunosorbent assay (ELISA) for antibodies to the ZEBRA product of EBV immediate early gene BZLF1. ZEBRA protein fused with glutathione-S-transferase (GST) was expressed in Escherichia coli and purified by affinity chromatography with glutathione-Sepharose 4B. An ELISA sandwich capture system was constructed with the GST-ZEBRA immobilized on plastic microtiter plates which had been coated with a mouse monoclonal antibody to GST. ZEBRA-IgG antibodies in patients' sera with chronic active EBV infection (CAEBV) and infectious mononucleosis (IM) had, respectively, very high and high titers. Anti-ZEBRA antibodies were also detected at low titers in sera of some healthy controls. ZEBRA-IgM antibodies were detected in sera of patients with IM and CAEBV but not in sera of healthy controls. In sera of patients with CAEBV, the titers of IgG antibodies to ZEBRA correlated with the antibody titers to early antigens obtained with an immunofluorescence assay, but not to EBV nuclear antigens. This ELISA is a useful diagnostic and prognostic test for EBV infection.

Adolescent↗