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Biomedical subjects

K Kuramoto

Publications and source records attributed to K Kuramoto.

At least 19 recordsLinked to original sources

Cell kinetics of hemopoietic colony-forming units in spleen (CFU-S) in young and old mice.

The growth potential of the hemopoietic progenitor cells from young and old mice is similar. Previous studies of their cell-kinetics showed no significant differences between them when measured by the incorporation of tritiated thymidine ([3H]TdR). A different approach for exploring stem-cell kinetics in aged animals is provided by another method; the incorporation of bromodeoxyuridine (BrdUrd) during DNA synthesis followed by exposure to near-ultraviolet light (near-UV) kills BrdUrd labelled cells in DNA-synthesis (S-phase). This BrdUrd-near UV cytocide (BUUV) reveals the size of cycling fractions at the level of hemopoietic progenitor cells; it also demonstrates the velocity of the cells entering S-phase when cells are labeled by a continuous infusion of BrdUrd by an osmotic pump, followed by an appropriate colony-assay for each progenitor cell. We compared the cell kinetics of young and old hemopoietic progenitor cells (CFU-S) by this approach. Osmotic pumps were implanted subcutaneously in the backs of young (2 months) and old (22 months) male C57BL/6CrSlc mice for 2, 4, 8, 12, and 16 days to continuously infuse BrdUrd at a flow-rate of 1 mg/h per kg. Cells were harvested from femoral marrow of the infused mice, plated in non-coated bacterial plates, and exposed to a single dose of near-UV at 4000 J/m2. After BUUV cytocide, bone-marrow cells were assayed for 8- and 13-day CFU-S colonies. In the 8-day colonies, the cytocide fraction of CFU-S from young mice increased rapidly, whereas the fraction from old mice showed flatter curves. Both curves reached a plateau at 52.6% for young, and 43.9% for old mice, and then converged 4 days after labeling. In the 13-day colonies, the curve for the aged was much flatter than that for the young; however, the plateaus in both young and old are similar, but at much lower values than earlier, i.e. 24.5% and 16.0%, respectively. The size of the cycling fraction of progenitor cells was close in the two groups during the steady-state cell-cycle. However, the velocity of the cell cycle at a progenitor level was very different, old mice being much slower. Further, within the progenitors, the cell cycle was much slower in the primitive ones and became faster when the stem cells differentiated into mature progenitor cells.

Aging

Hepatitis C virus infection in acquired aplastic anemia.

Hepatitis-associated aplastic anemia (HAAA) is an uncommon disorder that usually is not due to hepatitis A or B virus infection. Hepatitis C virus (HCV) seropositivity is infrequently observed in aplastic anemia (AA) patients who have not been extensively transfused. However, HCV seropositivity may not be detected until several weeks or months after viral infection and AA patients may exhibit defective humoral immunity. Therefore, we evaluated sera from AA patients for the presence of HCV viremia using a reverse transcriptase polymerase chain reaction (RT-PCR) based assay and several serologic assays for HCV antibodies. Serum samples from 90 AA patients who presented to the UCLA Medical Center between March 1984 and February 1990 were analyzed. Overall, 17 patients were found to have HCV viremia by RT-PCR assay, of whom 14 had a positive second-generation HCV enzyme immunoassay (EIA-2) and only 6 were EIA-1 reactive. The frequency of HCV viremia increased with the duration of time between diagnosis and sample procurement, and the number of blood products transfused prior to sampling (P = 0.026). No patient who received fewer than 20 U of blood products or who was sampled less than 20 days after diagnosis had a positive HCV RT-PCR result. Of four patients with hepatitis-associated AA (HAAA), one who was sampled 23 days after diagnosis had hepatitis C viremia and a reactive EIA-2 assay. Therefore, the high frequency of HCV viremia in this patient population is most likely due to transfusion with contaminated blood products prior to the introduction of routine blood donor screening for HCV.

Anemia, Aplastic

The effects of dilution on the outcome of pooled plasma testing with HIV type 1 (HIV-1) RNA genome amplification as compared to the outcome of individual-unit testing with other HIV-1 markers.

BACKGROUND: Proposed testing of large plasma pools with genome amplification technology (GAT) for detection of transfusion-transmissible viruses may have unanticipated complications not associated with individual unit testing. One such potential complication, the effect of dilution resulting from pool formation, was the subject of the present study. STUDY DESIGN AND METHODS: Specimens from three plasma donor HIV type 1 (HIV-1) seroconversion panels were tested with a quantitative HIV-1 RNA GAT assay (lower detection limit, 400 copies). GAT results were compared to HIV-1 p24 antigen and anti-HIV-1/2 enzyme immunoassay results. Effects of dilution on the detection of GAT-positive panel specimens were assessed by terminal dilution with pooled volunteer-donor EDTA plasma samples. RESULTS: Low HIV-1 RNA copy numbers (755 and 890 copies/0.1-mL input) that were detectable in two individual plasma specimens before HIV-1 p24 antigen were subsequently undetectable by GAT upon dilution with an equal volume of nonreactive plasma from a single donor. HIV-1 RNA at higher copy numbers (15,500 copies/0.1-mL input) in an HIV-1 p24 antigen-reactive and anti-HIV-1/2-nonreactive specimen was undetectable when diluted to 1-in-50 (1-in-50). Terminal dilution of seven HIV-1 RNA-containing plasma panel specimens indicated a proportional loss of HIV-1 RNA detectability with increasing dilution. CONCLUSION: GAT for detection of HIV-1 RNA in individual specimens was more sensitive than other HIV markers. For pooled plasma testing, GAT-independent constraints, such as effects of dilution, may preclude the use of GAT detection as a replacement for individual unit testing with HIV serologic assays.

Gene Amplification

An approach for acute disruption of large arteries in patients with advanced cervical cancer: endoluminal balloon occlusion technique.

OBJECTIVE: To test the feasibility of an intraluminal balloon occlusion technique for the control of sudden arterial disruption from cervical branches in patients with advanced malignancy. BACKGROUND: A sudden disruption of large cervical arteries is a devastating complication of advanced cervical malignancy and local infection. METHODS: Three patients with this complication underwent endoluminal balloon occlusion before surgical management. A double-lumen balloon catheter was introduced from the femoral artery to the bleeding point. Angiographic examination of the intracranial cross-filling could be performed by the injection of contrast medium from the opposite carotid artery, and consciousness levels could be directly confirmed under unilateral carotid occlusion. RESULTS: Resection of the carotid or innominate artery was safely performed in all the patients. No neurologic complications occurred. Bleeding did not recur during follow-up (range 5-32 months). CONCLUSIONS: This experience, although limited, suggests that surgical intervention to control cervical arterial bleeding with intraluminal balloon occlusion prevents excessive bleeding, decreases the risk of damage to the central nervous system, and improves the outcome in these critically ill patients.

Acute Disease

[Acquired pure red cell aplasia associated with relapsed non-Hodgkin's lymphoma: a case report-improvement of PRCA after acute hepatitis].

A 47-year-old male patient was admitted because of anemia. He had been diagnosed as non-Hodgkin's lymphoma (Follicular mixed, B cell type, stage ISA) by splenectomy two years before. Bone marrow examination on admission revealed lymphoma cell infiltration and marked decrease in erythroid cells. These findings confirmed relapsed lymphoma with acquired pure red cell aplasia. After several courses of combination chemotherapy, lymphoma cells disappeared from bone marrow, but PRCA was not improved. In this case there were two times remission of PRCA. At first time, acute B type hepatitis occurred during the chemotherapy, anemia improved transiently. At the second time, mild acute hepatitis associated with herpes zoster occurred. Twenty days after hepatic injury, PRCA was improved, and continued in remission state till present day. To disclose the mechanism of PRCA in this case, erythroid colony assay of marrow cells was performed. This showed the presence of inhibitory factor in patient's serum at PRCA state, that was considered to be related to the occurrence of PRCA. These findings suggest that the improvement of PRCA was associated with the changes on immunological condition after acute hepatitis in this case.

Acute Disease

Natural history of a spontaneous dissecting aneurysm of the proximal superior mesenteric artery: report of a case.

We report herein the case of a 44-year-old man in whom an asymptomatic dissecting aneurysm was found in the proximal part of the superior mesenteric artery (SMA) during a preoperative evaluation for colon cancer. The patient was managed conservatively with blood pressure control during the perioperative period of the colon resection as the false lumen of the dissecting aneurysm was revealed to be completely occluded by thrombus. The thrombus in the false lumen continued to be absorbed until 1 month after surgery. The patient is currently well 4 years after his operation without any evidence of recurrence of the aneurysm.

Adenocarcinoma, Papillary

Secretory granule formation and synthesis pathway of secretory proteins in parotid gland cells.

The cytochemical technique for endogenous peroxidase and an immunocytochemical method with protein A-gold were used to investigate the formation of the secretory granule substructure in acinar cells of the gerbil parotid glands in normal conditions and after treatment with the ionophore monensin. In untreated animals and 90 min after treatment with monensin, peroxidase activity was seen in rER, in transport vesicles close to the condensing vacuoles, and in the dense core of condensing vacuoles and secretory granules, whereas the Golgi cisternae remain unreactive. At 45 or 60 min after stimulation with monensin, the condensing vacuoles and the immature granules in the trans Golgi area did not exhibit dense cores and their content appeared similar to the peripheral portion of the mature secretory granules. These observations suggest that, in Mongolian gerbil parotid acinar cells, some substances such as peroxidase may be transported from the rER directly to the condensing vacuoles.

Animals

Reinitiation of spermatogonial mitotic differentiation in inactive old BDF1 mouse seminiferous tubules transplanted to W/Wv mouse testis.

The seminiferous epithelia of old mice (33 mo of age) are composed of spermatogonia and Sertoli cells. Histochemical examination using the anti-c-kit monoclonal antibody demonstrated that the number of differentiating type A spermatogonia decreases with age. To elucidate the differential activity of old mouse spermatogonia, we transplanted extremely thin seminiferous epithelia of old BDF, mice into W/Wv mouse testes and examined whether or not they could reinitiate differentiation. Artificially cryptorchid mice were used as the control. At 2 wk after transplantation, spermatocytes and round spermatids were detected in transplanted seminiferous tubules of the control, whereas the most advanced spermatogenic cells in those of old mice were spermatocytes. At 4 wk after transplantation, although elongated spermatids were detected in transplanted tubules of the control, haploid cells (spermatids) were still undetectable in those derived from old mice. Thus, meiosis was never restored, although spermatogonia of old mice can reinitiate differentiation into spermatocytes under suitable testicular conditions. Since it has been reported in several mammalian species that age-related changes in the testicular microenvironment lead to the gerontal cessation of spermatogenesis, the present results suggest that both a defective extratubular environment and a defective intratubular environment may cause the cessation of spermatogenesis in old BDF, mice.

Aging

Genotyping hepatitis C virus isolates from Spain, Brazil, China, and Macau by a simplified PCR method.

An improved and simplified method of genotyping was developed for classifying hepatitis C virus (HCV) isolates into the five common genotypes, i.e., I/1a, II/1b, III/2a, IV/2b, and V/3a, by PCR with genotype-specific primers deduced from the core gene. Sense and antisense primers, specific for each of the five common genotypes, were designed by comparison of 319 core gene sequences from HCV isolates of various genotypes from genetic groups 1 to 9. In the first round of PCR, a sequence of 433 bp representing nucleotides 319 to 751 was amplified with universal primers. The second round of PCR was performed with respective sense and antisense primers in two separate reactions, one for the amplification of genotypes I/1a and II/1b and the other for the amplification of genotypes III/2a, IV/2b, and V/3a. The specificity of genotyping was confirmed with a panel of 191 serum samples containing HCV isolates whose core gene sequences were known: 110 serum samples infected with HCV of the five common genotypes and 81 serum samples infected with HCV of other genotypes. The use of sense and antisense primers for genotype II/1b (primers 389 and 492) abolished the cross-reaction of the antisense primer for genotype II/1b (primer 133) with some HCV isolates of genotype I/1a found by our original method. The new method was used for genotyping 130 HCV isolates from Spain, 53 from Brazil, 106 from China, and 30 from Macau. A total of 329 bp of the NS5b region (nucleotides 8279 to 8607) of five isolates from Spain and five isolates from Macau which could not be classified as any of the five common HCV genotypes or genotype 2c were sequenced, and the sequences were compared with those of HCV isolates of known genotypes; two isolates from Spain were deduced to be of genotype 4d and one was deduced to be of genotype 1d, while the remaining two isolates from Spain had novel genotypes in genetic group 2; however, all five isolates from Macau were of genotype 6a.

Brazil

Hyaluronan and chondroitin sulfate in rabbit tears.

The concentrations of hyaluronan (HA) and chondroitin sulfate (CS) in rabbit tears were determined using high performance liquid chromatography. The tears were taken from both normal eyes and eyes with a corneal epithelial defect. The concentration of CS was 7.61 times higher than that of HA in the tears of normal eyes. When the concentration of CS isomers was compared, the highest concentration was observed in chondroitin 6-sulfate (C6S), followed by chondroitin 4-sulfate (C4S) and chondroitin (C0S), in that order. When the corneal epithelium was removed from the left eye using a trephine and microscissors, a significant increase was detected in the concentrations of HA and C6S in tears of the left eye one day after removal, whereas no change was observed in those of the right eye. In addition, a significant positive correlation was observed between the increased amount of HA concentration and the healing rate of the corneal epithelium. These results suggest that the increase of HA concentration in tears is associated with spontaneous corneal epithelial healing.

Animals

[Infection in elderly leukemic patients].

Febrile episodes occurring in 29 elderly patients (mean age 75 years) with leukemia, from 1988 to 1993, were reviewed. A febrile episode was defined as a temperature of 38 degrees C or greater for at least 6 hours. The number of febrile episodes was 64. The average was 2.2 febrile episodes per patient. Seventy-two percent of febrile episodes occurred when the patients had neutropenia below 100/microliters, while 16% occurred with neutropenia of 101/microliters to 500/microliters. Causative microorganisms were identified in 48% of total febrile episodes. The most common infectious site was the urinary tract which accounted for 25% of total episodes. Pneumonia and septicemia accounted for 22% of total episodes, respectively. Gram-positive cocci were responsible for 66% of microbiologically documented febrile episodes, while 21% were caused by gram-negative bacilli. Gram-positive cocci, particularly staphylococcus aureus, coagulae-negative staphylococcus and enterococci increased compared with a decade ago in our department. Granulocyte colony-stimulating factor (G-CSF) was used 12 times for infection. No significant difference in fever amelioration was seen between G-CSF and non-G-CFS cases.

Aged

[Long-term prognosis of chronic heart failure in the elderly].

To elucidate the predictors of the mortality rate in the elderly with chronic heart failure (HF), 120 consecutive patients (mean age, 75.2 +/- 7.8 years) with heart failure (NYHA I-II) were analyzed prospectively for 5 years. [Methods] Left ventricular ejection fraction (EF), left ventricular diastolic and systolic dimension (LVDD and LVDS) and wall thickness (WT) were measured by echocardiogram. Venipuncture for measurement of ANP and norepinephrine (NE) was done in supine position after 30 minute rest. [Results] 1) HF was associated with hypertension (47.5%), ischemic heart disease (34%), valvular disease (15%) and atrial fibrillation (AF, 23%). 2) 15 and 11 patients died for cardiac and non-cardiac events, respectively. 3) There was no difference in mean ages, gender, blood pressure, plasma-NE, EF, LVDD, LVDS, WT and AF between cardiac death and control groups. However, plasma ANP was higher in cardiac death group (173 pg/ml) than in control group (76 pg/ml) (p < 0.01). 4) Cox proportional hazard regression model revealed that ANP was an independent predictor for cardiac death (p < 0.005). We conclude that only plasma ANP level predicts long-term prognosis of chronic heart failure in the elderly.

Aged

[Primary myelofibrosis with severe bone pain].

An 80-year-old man was admitted with bone pain, affecting both lower limbs and the dorsa of the feet, in September, 1991. Primary myelofibrosis was diagnosed. Bilateral x-ray films of the tibia, fibula, femur and vertebra revealed osteoporosis, but this was considered an unlikely source of the bone pain. The bone pain subsequently disappeared, but then became severe in both lower limbs and the dorsa of the feet and was accompanied by leukemic transformations in February, 1992. He died of aspiration pneumonia in July, 1992. Autopsy revealed microfractures with osteoporosis and infiltration of leukemic cells in the periosteum of the left tibia, which may have caused the bone pain. This case provides useful information to analyze the cause of bone pain in patients with myelofibrosis.

Aged

Acute effects of exercise on muscle MRI in peripheral arterial occlusive disease.

The midcalf muscles of eight patients who had peripheral arterial occlusive disease were evaluated by exercise MRI before and after bypass surgery or percutaneous transluminal angioplasty. MRI showed a high intensity of these muscles, especially the posterior muscles, after exercise in all patients before intervention. The mean T2 relaxation time was maximal immediately after exercise (tibialis anterior, T2 = 30.8 ms; soleus, T2 = 36.2 ms; gastrocnemius, T2 = 32.8 ms) and then gradually decreased to the preexercise level. The difference in the T2 relaxation time of the soleus between immediately after exercise and at rest was smaller along with improvement of ankle pressure indices (API) after successful intervention (mean T2 difference: 4.91 and 0.72 ms (p < .001); mean API: 0.54 and 0.86 (p < .001) before and after intervention, respectively). The mean resting midcalf T2 relaxation time was significantly higher after intervention (tibialis anterior, T2 = 28.4 and 29.5 ms (p < .05); soleus, T2 = 31.4 and 32.9 ms (p < .05); gastrocnemius, T2 = 29.5 and 31.1 ms (p < .01) before and after intervention, respectively). T2 relaxation time may be a useful quantitative parameter in peripheral arterial occlusive disease as well as in other muscle studies.

Aged

Cardiac implications of the morning surge in blood pressure in elderly hypertensive patients: relation to arising time.

Although morning surge in blood pressure has been shown to be associated with the occurrence of myocardial ischemic events and stroke, few studies have been done regarding its pathogenesis, probably because of a lack of method for the quantitative assessment of awakening time. We conducted an echocardiographic study and ambulatory blood pressure monitoring in 23 elderly hypertensive patients to evaluate the relationship between the hypertensive cardiac change and morning surge in blood pressure. Of note was that the time of arising from bed was assessed quantitatively by an activetracer equipped with an internal acceleration sensor to monitor the physical activity. The change in systolic blood pressure after arising from bed was correlated significantly with the left ventricular mass index (r = 0.51, P < .02) and the A/E ratio, which represents the diastolic function (r = 0.70, P < .01). In contrast, the change in systolic blood pressure before rising from bed was not correlated with any echocardiographic parameters. We conclude that the magnitude of morning surge in blood pressure after arising from bed was related with the severity of hypertensive target organ damage.

Aged