Biomedical subjects
S Kojima
Publications and source records attributed to S Kojima.
Changes of SOD-like substances in mouse organs after low-dose X-ray irradiation.
We demonstrated that low-dose irradiation with 50 cGy of X-ray induces in vivo production of superoxide dismutase (SOD)-like substances and accelerates antioxidant activity. To elucidate the defense mechanism against X-ray radiation, we examined which components among these SOD-like substances, such as SOD, vitamin C and celuroplasmin, are produced by low-dose irradiation. Our study revealed that SOD-like specific activity hardly involved SOD-like substances other than SOD. Moreover, it is suggested that low-dose irradiation induced synthesis and production of SOD itself, leading to elevation of SOD-specific activity.
[Effect of the consumption of ethanol on the microcirculation of the human optic nerve head in the acute phase].
PURPOSE: The effect of the consumption of ethanol on the circulation of the optic nerve head (ONH) in the human eye in the acute phase and its mechanism were studied. METHODS: Eleven volunteers drank a bottle of beer (633 ml) with or without ethanol (29.5 g). Normalized blur (NB), a quantitative index of blood flow velocity, was measured in the temporal site of the ONH. NB, blood pressure (BP) and pulse rate (PR) were measured before, immediately after, and every 15 minutes for 90 minutes after consumption. Intraocular pressure (IOP) and plasma ethanol concentration were measured before, and 30 and 90 minutes after consumption. Genotyping of the aldehyde dehydrogenase (ALDH) 2 gene was also performed. RESULTS: NB in the ONH increased significantly from 15 to 45 minutes after consumption of ethanol and the maximum increase was 14% at 15 minutes. IOP was lowered at 90 minutes after consumption, but it was not significant. Mean BP was lowered significantly after 60 minutes. PR and ocular perfusion pressure did not change. A significant correlation was found between plasma ethanol concentration at 30 minutes and maximum NB. NB in the ALDH 2-deficient group was significantly larger from 15 to 45 minutes after consumption than in the proficient group. CONCLUSION: It appeared that the consumption of ethanol can increase the blood flow in the human ONH in the acute phase through decreased resistance in blood vessels induced by acetaldehyde, a metabolite of ethanol.
Use of granulocyte colony-stimulating factor for treatment of aplastic anemia.
Over the last ten years, recombinant human granulocyte colony-stimulating factor (rh G-CSF) has been widely used in the treatment of aplastic anemia (AA). It has been shown to facilitate the recovery of neutrophil count and useful for complicated bacterial or fungal infections. However, recent randomized clinical trials showed that the addition of rh G-CSF to immunosuppressive therapy had no clinical benefit for the prophylaxis of severe infections. These results suggested that rh G-CSF should be used for the treatment of infectious complications, not for the prophylaxis of infections in patients with AA.
[Effect of prednisolone on apoptosis and cellular infiltration in mdx mouse muscle].
Myonuclear apoptosis might precede the muscular degeneration of Duchenne muscular dystrophy. We examined influence of prednisolone (PSL) on apoptosis in the skeletal muscle of mdx mouse. Amount of apoptotic nuclei of the tibial anterior or quadriceps muscle was assessed by Tdt-mediated dUTP biotin nick end-labeling (TUNEL) method. When nuclei were classed into myonuclei and interstitial ones, apoptotic nuclei were observed mainly in the latter. In comparison between 7 and 33 week-old mice, interstitial apoptosis increased with age in both control and PSL group. Treatment with PSL decreased number of interstitial apoptotic nuclei. To the contrary, apoptotic myonuclei were increased with PSL treatment. Treatment with PSL significantly reduced focal accumulation of infiltrating mononuclear cells, possibly contributing to the suppression of muscle degeneration. Relationship between decrease of interstitial apoptosis and reduced cellular infiltration was unclear yet. Present study showed that PSL differently affected apoptosis between muscle cells and their interstitium.
Health education and community participation in the control of urinary schistosomiasis in Ghana.
OBJECTIVE: To study the role of health education and community participation for the provision of facilities necessary for the control of urinary schistosomiasis in southern Ghana. HYPOTHESIS: Health education facilitates community participation in the provision of facilities for the control of bilharzia. STUDY AREA: Three rural communities drained by the Densu river in southern Ghana. PARTICIPANTS: Individuals aged 14 years and above formed groups of 10-12 persons by age, sex, ethnic and educational background; 15-16 groups were formed. INTERVENTIONS: Based on existing structures, one community received active, another passive health education and the third had no education. All three communities received chemotherapy. DESIGN: Study was carried out in three phases: pre-intervention phase--during which baseline data on residents' knowledge, attitude, beliefs and perception about bilharzia were collected using focus group discussions (FGD) prior to the second phase, intervention. Another FGD was held after 18 months to evaluate the intervention--third phase. RESULTS: This study suggests that most community members were aware of schistosomiasis but not as a disease. Before the health education, some residents believed bilharzia was a sign of manhood while others attributed the red colour of the urine to the red colour of a variety of sugar cane eaten in the area. After the health education, residents in the three areas constructed hand-dug wells. In addition, those who received active health education constructed two toilets for the schools and weeded the banks of the rivers. Residents also associated the disease with the water snail. CONCLUSION: Health education was useful in changing community perception on bilharzia.
Localization of glutathione and induction of glutathione synthesis-related proteins in mouse brain by low doses of gamma-rays.
First, we determined the cerebral localization of reduced glutathione (GSH) in normal mice by means of autoradiography using 99mTc-meso-hexamethyl propylene oxime. A highly specific localization of GSH in the cerebellum and hippocampus was observed. Secondly, we measured the elevation of GSH level in the brain after low-dose gamma-irradiation. The cerebral GSH levels increased soon after irradiation with 50 cGy of gamma-rays, reaching a maximum at 3 h post-treatment, then remaining significantly higher than that of the non-irradiated control until 12 h and returning to the control level by 24 h. Thirdly, we examined the induction of the activities and the mRNAs of proteins involved in the synthesis and regeneration of GSH in the brain of mice subjected to low-dose gamma-ray irradiation. The level of mRNA for gamma-glutamylcysteine synthetase was significantly increased at 0.5 h, and remained high until 2 h post-irradiation (50 cGy). The level was transiently lowered to the non-irradiated control level at 3 h and slightly increased again after 6 h post-irradiation. gamma-Glutamylcysteine synthetase activity was significantly increased 3 h after irradiation, and remained high up to 24 h post-irradiation. As for glutathione reductase, the mRNA level was increased at 0.5 h, and peaked strongly at 2 h, while the enzyme activity was significantly increased at 6 h after irradiation, and continued to increase up to 24 h. The level of mRNA for thioredoxin, which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked between 0.5 h and 2 h post-irradiation, and rapidly declined thereafter. The content of thioredoxin showed a transient decrease immediately after irradiation, but was then remarkably elevated, reaching a maximum at 3 h, and thereafter declining sharply. These results indicate that the increase in endogenous GSH in mouse brain soon after low-dose gamma-ray irradiation is a consequence of the induction of GSH synthesis-related proteins and occurs via both the de novo synthesis and the regeneration pathways.
Sensory preconditioning for feeding response in the pond snail, Lymnaea stagnalis.
We demonstrated a sensory preconditioning in the pond snail, Lymnaea stagnalis. An appetitive sucrose solution (a conditioned stimulus: CS1) and weak vibration (another conditioned stimulus: CS2) were first associated, and then the CS2 and an aversive KCl solution (an unconditioned stimulus: UCS) were done. To build the conditioning, two different training procedures, spaced and massed, were examined. After the both training, the sensory preconditioning was built: significantly fewer feeding response to the CS1 became elicited; slower latency to the first bite to the CS1 was induced. No significant differences on the memory retention between these training procedures were found in the sensory preconditioning.
Prediction of human herpesvirus 6 infection after allogeneic bone marrow transplantation.
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Organization and structure of NADH-dependent glutamate synthase gene from rice plants.
Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (lambdaOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (lambdaOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (lambdaOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH4Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH-GOGAT to identical positions. The 3. 7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database.
Induction of mRNAs for glutathione synthesis-related proteins in mouse liver by low doses of gamma-rays.
We examined the elevation of the reduced form of glutathione (GSH)level and the induction of MRNAs for proteins involved in the synthesis and regeneration of GSH in the liver of mice after low-dose gamma-ray irradiation. The liver GSH level increased soon after irradiation with 50 cGy of gamma-rays, reached a maximum at around 12 h post-treatment. The mRNA of gamma-glutamylcysteine synthetase (gamma-GCS), the rate-limiting enzyme for de novo synthesis for GSH, showed a small increase that peaked at 6 h after gamma-ray irradiation at a dose of 50 cGy. Only a small increase in gamma-GCS activity was observed throughout the 24-h post-irradiation period. In the case of glutathione reductase (GR), which is involved in the regeneration of GSH from the oxidized form (GSSG), the mRNA level peaked strongly at 1 h, while the activity peaked at twice the control level 12 h after irradiation. The level of mRNA for thioredoxin (TRX), which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked at 1 h and declined thereafter, while the activity peaked at 3 h and then declined sharply. These results indicate that the increase in endogenous GSH immediately following low-dose gamma-ray irradiation is predominantly due to operation of the regeneration cycle and not de novo synthesis. We also examined the dependence of mRNA induction on the gamma-ray dose.
Facilitation by endogenous acetylcholine and nitric oxide of luminal serotonin release from the guinea-pig colon.
The present study was designed to determine the influence of endogenous acetylcholine and nitric oxide (NO) on spontaneous luminal serotonin (5-hydroxytryptamine, 5-HT) release in the luminally perfused isolated guinea-pig proximal colon in vitro. 5-HT was determined by high-performance liquid chromatography with electro-chemical detection. The luminal outflow of 5-HT was significantly reduced by atropine (0.2 microM), hexamethonium (100 microM), the NO synthase inhibitor NG-nitro-L-arginine (L-NNA, 10 microM) and the NO-trapping agent 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO, 30 microM). Addition of excess L-arginine (300 microM) reversed the inhibitory effect of L-NNA on the 5-HT outflow. Physostigmine (1 microM) caused a great increase (atropine-sensitive) in 5-HT outflow. The enhancing action of physostigmine on 5-HT outflow was partially inhibited by L-NNA (100 microM) or carboxy-PTIO (30 microM), but was unaffected by the muscarinic M1 receptor antagonist pirenzepine (0.2 microM) or a muscarinic M3 receptor antagonist 4-diphenyl-acetoxy-N-methyl-piperidine methiodide (0.2 microM). These results suggest that 5-HT release from luminally perfused proximal colon of the guinea pig is stimulated via a NO pathway and cholinergic pathways which utilize muscarinic synapses and nicotinic synapses. Further, an intrinsic cholinergic-NO link appears to play a role in the stimulation of luminal 5-HT release, which may reflect the release of 5-HT from entero-chromaffin cells.
Improved restriction landmark cDNA scanning and its application to global analysis of genes regulated by nerve growth factor in PC12 cells.
Restriction landmark cDNA scanning (RLCS) is a novel method by which more than 1000 genes can be simultaneously and quantitatively displayed as two-dimensional gel spots. Here we present an adaptation that allows an individual spot to correspond to a unique gene species without redundancy in more than two gel patterns. Using this improved RLCS, we examined global changes on the gene expression of PC12 cells before and after treatment with nerve growth factor. Among a total of 3000 spots, 21 (0.70%) and 91 (3.03%) spots newly appeared and became more intense with treatment. On the other hand, 15 (0.50%) and 44 (1.47%) spots disappeared, becoming less intense with treatment. These observations suggest that approx. 6% of the detected PC12 genes are up-(3.73%) or down-(1.97%) regulated when the cells differentiate to neuronal cells. In comparison with the results obtained using the expressed-sequence-tag approach, previously reported by Lee et al. (Proc. Natl. Acad. Sci. USA 92 (1995) 8303-8307), RLCS should be useful for quantitatively examining the global change of differentially expressed genes of various expression levels.
Change of glutathione peroxidase synthesis along with that of superoxide dismutase synthesis in mice spleens after low-dose X-ray irradiation.
We have previously demonstrated that the activity of superoxide dismutase (SOD), an antioxidant, is enhanced by low-dose X-ray irradiation in various organs of animals such as rats. Since SOD is an enzyme that mediates the dismutation of O2- to H2O2, the question as to whether the resultant H2O2 is further detoxicated into H2O and O2 or not must still be evaluated. Hence, we studied the effect of low-dose X-ray irradiation on the synthesis of glutathione peroxidase (GSHPx), which is an antioxidant that catalyzes this reaction. The results suggest that H2O2 produced by increased SOD activity can be detoxicated into H2O and O2 due to simultaneous enhancement of the GSHPx activity by X-ray irradiation at 20 cGy, in contrast to irradiation at 400 cGy. The results also show the enhancement in enzyme activities by induction of their synthesis shortly after irradiation at 20 cGy. Moreover, as this phenomenon was observed in BALB/c mice (which are more radiation-sensitive compared to other mouse strains) and radiation-resistant C57BL/6NJcl mice, it was considered to be a common phenomenon in the rat spleen.
Functional transitions in myosin: formation of a critical salt-bridge and transmission of effect to the sensitive tryptophan.
For analyzing the mechanism of energy transduction in the "motor" protein, myosin, it is opportune both to model the structural change in the hydrolytic transition, ATP (myosin-bound) + H2O --> ADP.Pi (myosin-bound) and to check the plausibility of the model by appropriate site-directed mutations in the functional system. Here, we made a series of mutations to investigate the role of the salt-bridge between Glu-470 and Arg-247 (of chicken smooth muscle myosin) that has been inferred from crystallography to be a central feature of the transition [Fisher, A. J., Smith, C. A., Thoden, J. B. , Smith, R., Sutoh, K., Holden, H. M., & Rayment, I. (1995) Biochemistry 34, 8960-8972]. Our results suggest that whether in the normal, or in the inverted, direction an intact salt-bridge is necessary for ATP hydrolysis, but when the salt-bridge is in the inverted direction it does not support actin activation. Normally, fluorescence changes result from adding nucleotides to myosin; these signals are reported by Trp-512 (of chicken smooth muscle myosin). Our results also suggest that structural impairments in the 470-247 region interfere with the transmission of these signals to the responsive Trp.
Tertiary structure formation in the propeptide of subtilisin BPN' by successive amino acid replacements and its close relation to function.
The propeptide of subtilisin BPN', located between a signal peptide and the mature region of the protease, is known to exhibit inhibitory activity toward subtilisin BPN', in addition to its activity as an intramolecular chaperone that facilitates folding of subtilisin BPN'. Another unique feature is that although the isolated propeptide is in a random-coil state, it forms a defined tertiary structure when it is bound to subtilisin BPN'. In this study, amino acid replacements likely to increase the hydrophobicity of the propeptide have been introduced so that the isolated propeptide forms a defined tertiary structure. By successive replacements of Ala47 by Phe, Gly13 by Ile and Val65 by Ile, the propeptide was found to form a tertiary structure in addition to an increase in its secondary structure content, which were identified by circular dichoism spectra measurements. Concurrently, the propeptide, which is a temporary inhibitor in its wild-type form, became resistant to proteolytic digestion by subtilisin BPN'. These results show not only the close relationship between tertiary structure formation in the propeptide and its function as a protease inhibitor but also the ability of a random-coil protein to form a tertiary structure after a limited number of well-designed amino acid replacements.
Elevation of the serum Fas ligand in patients with hemophagocytic syndrome and Diamond-Blackfan anemia.
Fas ligand (FasL) is a membrane protein that is expressed in activated T cells and natural killer cells. FasL binds to Fas on target cells and induces apoptosis. There exists a soluble form of FasL (sFasL), and sFasL also induces apoptosis of Fas-bearing cells. The serum sFasL concentrations were reported to be elevated in patients with large granular lymphocytic leukemia and natural killer cell lymphoma. In this study, we have measured serum sFasL concentrations in other hematological disorders, including severe aplastic anemia (SAA), hemophagocytic lymphohistiocytosis (HLH), and Diamond-Blackfan anemia (DBA). The serum sFasL concentration of age-matched healthy controls was 0.16 +/- 0.11 ng/mL (mean +/- SD, n = 22). The serum sFasL levels in the patients with HLH and DBA were 3.75 +/- 3.82 (n = 19; P < .0001, HLH v control) and 2.76 +/- 2.43 ng/mL (n = 6; P = .012, DBA v control), respectively. Serum interferon-gamma concentration was elevated in the patients with HLH (1.61 +/- 2.62 ng/mL) but not in those with DBA (below the detectable level). These results suggest that the Fas-FasL system plays a role, at least in part, in the pathophysiology of HLH and DBA.
Plasma natriuretic peptides as indicators of left ventricular remodeling after myocardial infarction.
To investigate the relationship between natriuretic peptides and left ventricular remodeling after acute myocardial infarction, left ventriculography and blood sampling were performed on admission, after 1 month and after 3 months in 33 patients with acute myocardial infarction (15 anterior and 18 inferior). Plasma atrial and brain natriuretic peptide concentrations at 1 and 3 months were higher than those of controls (P<0.01). Brain natriuretic peptide concentrations correlated with changes in left ventricular end-diastolic volume index after 1 and 3 months (1 month: r=0.57, P=0.003; 3 months: r=0.47, P=0.006). Atrial natriuretic peptide concentrations also correlated with changes in left ventricular end-diastolic volume index after 1 and 3 months (1 month: r=0.40, P=0.02; 3 months: r=0.61, P<0.001). Our results indicate that natriuretic peptide concentrations increase in the chronic phase of acute myocardial infarction and may relate to left ventricular remodeling. Thus, atrial natriuretic peptide as well as brain natriuretic peptide concentrations may be useful biochemical markers in identifying asymptomatic patients at risk for heart failure or sudden death after acute myocardial infarction.