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Biomedical subjects

M Takayanagi

Publications and source records attributed to M Takayanagi.

At least 127 records · Page 7Linked to original sources

[Efficacy of early use of continuous isoproterenol inhalation therapy for severe asthma attacks in children].

The aim of this study was to evaluate the efficacy and safety of continuous isoproterenol inhalation therapy for asthma attacks in children. We used l-body isoproterenol (Proternol L) in 22 children with 32 episodes of severe attacks. One of them did not respond to this therapy, and two had complications (atelectasis and pneumothorax). Twenty-nine cases were divided into three subgroups according to their clinical scores; A) scores less than or equal to 4, which meant that they were in the early stage of severe attack (n = 9), B) scores 5-6, which meant impending respiratory failure (n = 17), C) scores greater than or equal to 7, which meant respiratory failure (n = 3). The values of SpO2 at the start of this therapy were 94.8, 91.5, 82.0%, respectively. The more severe their attacks were, the lower their SpO2 levels were. The periods until their scores became zero were 0.78, 6.3, 17.2 hours, respectively. There were significant differences between each period respectively (p less than 0.001, p less than 0.01). Heart rates decreased when their symptoms improved, and other adverse effects were not detected. These results suggest that this therapy is effective and safe for children with severe asthma attacks, especially in the early stage.

Administration, Inhalation↗

[Efficacy of continuous isoproterenol inhalation therapy for severe asthma attacks in younger children].

The aim of this study was to evaluate the efficacy of continuous isoproterenol inhalation therapy for severe asthma attacks in younger children, compared with its efficacy in older children. We used l-body isoproterenol (Proternol L) in 31 children with 42 episodes of severe attacks. They were divided into two group according to age: 20 cases under 6 years old (Group A), and 22 cases over 7 years old (Group B). All of the patients except for one in Group B, eventually improved with this therapy. Wood's clinical scores for Group A were significantly higher than those for group B (p < 0.01). In 22 cases whose scores were 5-6, their SpO2 values at the onset of this therapy were 90.8 +/- 3.17 in group A and 92.4 +/- 3.82% in group B. The improvement time of group A (13.6 +/- 16.2 hours) was significantly longer than that of group B (2.5 +/- 5.66, p < 0.01). The nebulized isoproternol doses for group A were 0.47 +/- 0.168 and for group B 0.26 +/- 0.096 mg/kg/saline 500 ml. The dose for group A was significantly higher than that for group B (p < 0.01). We concluded that continuous isoproterenol inhalation therapy was effective even in younger children. But the degree of efficacy was slightly lower in younger children, although they inhaled higher doses of isoproterenal than older children.

Administration, Inhalation↗

Apoptosis in cultured rat hepatocytes: the effects of tumour necrosis factor alpha and interferon gamma.

We investigated the cytotoxic effects of tumour necrosis factor alpha (TNF alpha) and interferon gamma (IFN gamma) on rat hepatocytes in culture. Under phase contrast microscopy, we found a small number of dying hepatocytes in control cultures, each having been transformed into a cluster of small spheres. Under transmission electron microscopy, these cells showed the characteristics of apoptosis. TNF alpha and a combination of TNF alpha and IFN gamma exerted a cytotoxic effect, whereas IFN gamma showed no significant cytotoxicity when assessed by neutral red assay and by measuring LDH activity in culture medium. Under phase contrast microscopy, the number of apoptotic cells increased with the addition of either TNF alpha or IFN gamma, and markedly with the addition of both. DNA extracted from apoptotic cells cultured with TNF alpha and IFN gamma was fragmented, and a set of bands of the '200 bp ladder', which is characteristic of the DNA of apoptotic cells, was observed in agarose gel electrophoresis. These findings indicate that cultured hepatocytes die from apoptosis. TNF alpha killed cultured rat hepatocytes by increasing apoptosis, and this effect was potentiated by the addition of IFN gamma, which by itself was also weakly cytotoxic.

Animals↗

Activity of antibodies to hepatitis C virus of patients with chronic non-A, non-B hepatitis decreases during interferon alpha therapy.

The activity of antibodies to hepatitis C virus (anti-HCV) was investigated in 80 patients with chronic non-A, non-B liver diseases. Anti-HCV antibodies were positive in 82.5% (66/80), and the titers were 1 for 18 patients, 2 for 40 and 3 for 8, respectively. The frequency of anti-HCV was significantly lower in patients with chronic persistent hepatitis (8/13, 61.5%) than in those with chronic active hepatitis (42/49, 85.7%) (P less than 0.05). There was no significant difference in the distribution of anti-HCV titers among the different stages of hepatitis. There was no correlation between anti-HCV titer and histology activity index score in chronic hepatitis. Activity of anti-HCV decreased more frequently in the patients who responded to interferon alpha (IFN alpha) therapy (8/22, 36.4%) than in those who did not (0/9, 0%) (P less than 0.05). These results indicate that anti-HCV activity does not correlate with the activity or disease stage of chronic hepatitis, but that anti-HCV activity decrease more frequently during IFN alpha treatment in patients who responded to IFN alpha therapy.

Adult↗

CO2 laser-assisted microvascular anastomosis: biomechanical studies and clinical applications.

Experiments have been performed using CO2 laser-assisted microvascular anastomoses, and they demonstrated the following features, in comparison with conventional anastomoses: ease in technique; less time consumption; less tissue inflammation; early wound healing; equivalency of patency rate and inner pressure tolerance; but only about 50 percent of the tensile strength of manual-suture anastomosis. Six clinical applications in 16 vessels are reported, using this procedure. The preliminary results of these cases would appear to be the first successful replantations and free tissue transfers using CO2 laser-assisted microvascular anastomoses in man. The procedure offers increased safety and speed in microvascular anastomoses.

Adolescent↗

Effective prophylaxis of thrombosis by antithrombin III concentrate in a pregnant woman with congenital antithrombin III deficiency: relations between plasma antithrombin III activity and the plasma levels of hemostatic molecular markers.

The value of antithrombin III (AT III) concentrate and a standard criterion for its use were examined in a pregnant woman with congenital AT III deficiency by continuous monitoring of plasma AT III activity and the plasma levels of hemostatic molecular markers. The rates of improvement of various markers after AT III administration (frequency of improvement/frequency of administration) were as follows: fibrinopeptide A (FPA) 82%, D-dimer 70%, fibrinopeptide B beta 15-42 73%, beta-thromboglobulin 60%, and platelet factor 4 50%. There was no significant correlation between the plasma AT III activity and all plasma FPA values, but FPA values of over 3.9 ng/ml showed a significant negative correlation with AT III activity: AT III activity (%) = -6.59 x FPA (ng/ml) + 125, r = -0.851, p less than 0.02. We therefore recommend continuous monitoring of the plasma FPA level and administration of AT III concentrate when the FPA level is elevated. According to the regression line shown above, plasma AT III activity should be raised to 100% to keep the FPA level below 6.0 ng/ml with 95% confidence limit.

Adult↗

Detection of pre-C and core region mutants of hepatitis B virus in chronic hepatitis B virus carriers.

We analyzed the pre-C and core region of hepatitis B virus (HBV) DNA by a polymerase chain reaction in 22 chronic carriers. In 9 hepatitis B e antigen-positive asymptomatic carriers, a single DNA band was detected at the expected size, whereas additional shorter DNA bands were observed in 7 out of 11 patients with chronic hepatitis. The smaller-sized DNAs from one chronic hepatitis patient had various lengths of deletions spanning from 105 to 183 bp in the middle of the core gene, and all deletions included common nucleotide sequences. All of the smaller-sized DNAs from the other patients proved to be variant core genes. They were deleted in similar regions by Southern analysis using oligonucleotide probes. A follow-up study revealed that four out of seven chronic hepatitis patients with a short core gene seroconverted to antibody to hepatitis B e antigen, but those with only a "wild type" did not. In another set of sequence studies, clones isolated from two chronic carriers displayed heterogeneity of the pre-C and core gene which was more often present in sera with normal alanine aminotransferase levels than with abnormal levels. These results suggest that mutant HBV alters the host immune response, and may modulate the clinical course of HBV infection. An alternative possibility is that chronic hepatitis selects for mutant forms.

Adolescent↗

Use of the o-phenylenediamine fluorescence system in the enzymatic assay of serum uric acid.

A manual enzymatic method is described for sensitive fluorometric determination of uric acid in human serum. This method is based on an enzymatic reaction with uricase to form hydrogen peroxide from uric acid and the following oxidation of o-phenylenediamine with peroxidase and hydrogen peroxide for the production of a fluorescence compound. The specificity and the selectivity in the method are due to the uricase reaction and the fluorometry, respectively. The formed fluorescence in the reaction mixture is measured at 410 nm (an excitation) and 550 nm (an emission). This enzymatic method can determine uric acid at 30-1000 microM, with a between-assay relative standard deviation of 4.35% or less. A good correlation is obtained between the present method and the colorimetric kit method.

Ascorbate Oxidase↗

Basic fibroblast growth factor and epidermal growth factor promote survival of primary cultured cerebellar neurons from neonatal rats.

Neurotrophic effects of CS23, a mutein of human basic fibroblast growth factor (bFGF), and recombinant human epidermal growth factor (hEGF) were compared in dissociated cerebellar cultures from embryonic (E20) and early postnatal (P0-5) rats. Both CS23 and hEGF promoted the survival of cerebellar neurons without distinction of ages of animals used. The maximum effects of CS23 were always larger than those of hEGF. These results suggest that both bFGF and EGF also function as neurotrophic factors in postnatal cerebellum.

Animals↗

Involvement of protein kinase activation in neurotrophic effects of basic fibroblast growth factor in cultured brain neurons.

The influences of K-252a and staurosporine, protein kinase inhibitors, on neurotrophic effects of basic fibroblast growth factor (bFGF) were investigated in dissociated cell cultures of the striatum, hippocampus and cerebellum of fetal rats. Addition of 1 ng/ml bFGF enhanced the survival of cultured neurons of all brain regions tested. Both K-252a (10-200 nM) and staurosporine (1-100 nM) blocked the survival promoting effects of bFGF in a concentration-dependent manner. These results suggest that bFGF exerts its neurotrophic effects through activation of protein kinase(s).

Alkaloids↗

In vitro production of antimitochondrial antibody by peripheral blood mononuclear cells in patients with primary biliary cirrhosis.

Peripheral blood mononuclear cells from patients with primary biliary cirrhosis (PBC) were studied for their ability to produce antibodies to mitochondrial antigens (antimitochondrial antibody, AMA and anti-pyruvate dehydrogenase complex, anti-PDC), using dot-immunobinding and ELISA assay. 2 x 10(6) B cells alone and the mixture of 1.4 x 10(6) T plus 0.6 x 10(6) B cells from patients with serum AMA+ PBC spontaneously produced similar amounts of AMA and anti-PDC. Coculture experiments of T and B cells showed that anti-PDC production was significantly greater in pokeweed mitogen (PWM)-stimulated cultures than in unstimulated ones. The reactivities of antibodies to mitochondrial antigens were inhibited by M2 mitochondrial antigen-enriched fraction. Ursodeoxycholic acid added to cultures resulted in a significant inhibition for production of immunoglobulins and anti-PDC (IgG and IgM). Spontaneous production of AMA or anti-PDC was not detected in patients with AMA- PBC, chronic viral hepatitis and healthy controls, nor could it be induced by PWM in either group. EBV-transformed B cell line established from a AMA+ PBC patient produced high levels of AMA and IgM class anti-PDC. These findings indicate that antibody to a specific mitochondrial antigen in patients with PBC may be produced in T cell-dependent and-independent manner.

Adult↗

Anti-hepatitis C antibodies in patients with chronic non-A, non-B hepatitis: relation to disease progression and effect of interferon alpha.

The activity of antibodies to hepatitis C virus (anti-HCV) was investigated in 80 patients with chronic non-A, non-B liver diseases. Serum anti-HCV titer was determined by the "Ortho-HCV" enzyme-linked immunosorbent assay with some modifications to quantify the activity. Anti-HCV was positive in 82.5% of cases (66/80). Anti-HCV occurred significantly less often in the patients with chronic persistent hepatitis (8/13, 61.5%) than in those with chronic active hepatitis (42/49, 85.7%) (p less than 0.05). Anti-HCV titer of the patients with chronic persistent hepatitis and that with chronic active hepatitis was significantly higher than that with liver cirrhosis (p less than 0.01 and p less than 0.05). There was no correlation between anti-HCV titer and histology activity index in chronic hepatitis. Serial study demonstrated that anti-HCV titer decreased more frequently in the patients who responded to IFN alpha therapy (11/22, 50.0%) than in those who did not respond to IFN alpha therapy (0/10, 0%) (p less than 0.01). These results indicate that anti-HCV level does not correlate with the activity of hepatitis, but that it decreases in accordance with the disease progression to liver cirrhosis or with the response to IFN alpha therapy.

Adult↗

Serum interleukin 6 levels in patients with chronic hepatitis B.

Studies were undertaken to evaluate the role of interleukin 6 (IL-6) in the pathogenesis of chronic hepatitis B. Using recently developed sensitive and specific enzyme-linked immunosorbent assay, we determined serum levels of IL-6 in patients with chronic active hepatitis B (B-CAH), chronic persistent hepatitis B (B-CPH), and primary biliary cirrhosis (PBC). Serum IL-6 activity tended to increase in patients with B-CAH (38.4 +/- 68.0 pg/ml) (+/- SD), compared with controls (9.7 +/- 6.8 pg/ml), although the difference was not significant because of the wide scatter in values. IL-6 levels in B-CPH and PBC were similar to those of controls. There was a significant correlation between the levels of IL-6 and aminotransferase in B-CAH. In addition, IL-6 activity was significantly enhanced during acute exacerbation of the illness, accompanied by clearance of HBeAg. Patients who received IFN-alpha and IL-2 therapy showed depressed IL-6 activity during treatment, and a rebound beyond pretreatment values after cessation of therapy. These findings suggest that IL-6 plays a role in the development of chronic hepatitis B, and it may contribute, at least in part, to the elimination of HB virus.

Adult↗

Effects of recombinant human basic fibroblast growth factor and its modified protein CS23 on survival of primary cultured neurons from various regions of fetal rat brain.

Neutrophic effects of recombinant human basic fibroblast growth factor (hbFGF) and its modified protein CS23 on brain neurons were evaluated by their abilities to promote survival of primary cultured neurons from various regions of fetal rat brain. In the molecule of CS23, two serines are substituted for two cysteines at positions 70 and 88 in the natural hbFGF. Both hbFGF and CS23 markedly increased the survival of cultured neurons from all regions tested, i.e., cerebral cortex, septum, striatum, hippocampus, thalamus, substantia nigra, colliculus and cerebellum. The effects were concentration-dependent, in the range of 0.01 to 10 ng/ml, in similar manners among all regions. Although the maximal number of surviving neurons in the presence of CS23 was little different from that of hbFGF, CS23 could significantly promote neuronal survival at a lower concentration than hbFGF. The time-course of these effects on survival was almost the same between hbFGF (1 ng/ml) and CS23 (1 ng/ml). These results suggest that hbFGF has a strong neurotrophic activity on a wide range of brain neurons and that CS23 maintains the activity of original hbFGF successfully.

Animals↗

A comparison of neurotrophic effects of epidermal growth factor and basic fibroblast growth factor in primary cultured neurons from various regions of fetal rat brain.

Effects of recombinant human epidermal growth factor (hEGF) on survival of primary cultured neurons from various regions of fetal rat brain were compared with those of recombinant human basic fibroblast growth factor (hbFGF). Addition of hEGF (0.001-10 ng/ml) in chemically-defined serum-free medium enhanced significantly the survival of cultured neurons from all regions tested, i.e., cerebral cortex, septum, striatum, hippocampus, thalamus, colliculus and cerebellum. However, its effective concentration differed considerably with the regions. The most sensitive regions were the colliculus and cerebellum (minimum effective concentration, 0.001 ng/ml), while the least sensitive was the cerebral cortex (minimum effective concentration, 10 ng/ml). Maximum numbers of surviving neurons in the presence of hEGF were smaller than those of hbFGF in all regions. Maximum effects of hEGF compared with those of hbFGF was greatest in the striatum and smallest in the cerebral cortex. These results suggest that hEGF may exert neurotrophic effects for limited populations of brain neurons. When hEGF (1-10 ng/ml) and hbFGF (0.01-10 ng/ml) were added together, hEGF increased the effects of a lower concentration of hbFGF in an additive manner, but the maximum effects of hbFGF were not potentiated by hEGF in all regions tested. Therefore, mechanisms underlying the effects of hEGF very probably overlap with those of hbFGF.

Animals↗

Determination of acetaldehyde in human blood by high-performance liquid chromatography using fluorometry.

A method was developed for the measurement of acetaldehyde in human blood by high-performance liquid chromatography (HPLC). The method was based on a pre-column reaction; a fluorescent substance was formed by a coupling reaction between 2 mol of cyclohexan-1,3-dione and 1 mol of acetaldehyde with ammonium acetate. The coupling compound was analyzed by HPLC and the concentration of acetaldehyde was obtained from the calibration curve drawn from the results using standard solutions. The determination was sensitive and reproducible with a range of 0.2-10 microM and a precision (coefficient of variation) of 2.43%.

Acetaldehyde↗