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Biomedical subjects

M Yamaguchi

Publications and source records attributed to M Yamaguchi.

At least 901 records · Page 50Linked to original sources
PubMed1994

Memory deficit accompanying cerebral neurodegeneration after stroke in stroke-prone spontaneously hypertensive rats (SHRSP).

Memory performances of SHRSP with chronic stroke were examined on the three-panel runway task in addition to the histological evaluation and magnetic resonance imaging (MRI) of the brain. After recovery from the neurological symptoms with stroke. SHRSP were subjected to acquisition training on the memory tasks, and they exhibited both a delay and a persistent impairment of acquisition on the memory tasks, compared to the non-stroke SHRSP. T2-weighted MRI with the stroke SHRSP suggested marked edematous formation in the cortex, caudate putamen and/or thalamus, preferentially in the frontal and/or occipital cortex. The histological evaluation showed edematous degeneration such as edema, gliosis and cyst preferentially in the cortex, but no degeneration in the hippocampus. Thus, SHRSP with chronic stroke was found to exhibit impairment of learning and memory, which may be due to the cortical edematous degeneration.

Animals↗
PubMed1994

Differential effects of insulin and insulin-like growth factor-I in the femoral tissues of rats with skeletal unloading.

The alteration of bone metabolism in the femur of rats with skeletal unloading for 4 days was investigated. Skeletal unloading was designed using the model of hindlimb hang in rats. Skeletal unloading caused a significant decrease in femoral weight, calcium, and phosphorus contents in the metaphysis but not diaphysis. Also, the unloading induced a significant decrease of zinc content, alkaline phosphatase activity, and deoxyribonucleic acid (DNA) content in the femoral diaphysis and metaphysis. When the femoral-diaphyseal and metaphyseal tissues from normal and skeletal-unloading rats were cultured in the presence of insulin (10(-9) and 10(-8) M) for 24 hours in vitro, the hormonal effect to increase alkaline phosphatase activity and DNA content in the diaphysis, but not metaphysis, was lost in the bone tissues from unloading rats. However, the culture with insulin-like growth factor-I (IGF-I; 10(-8) and 10(-7) M) produced a significant increase of alkaline phosphatase activity and DNA content in both the diaphyseal and metaphyseal tissues from normal and unloading rats. These results demonstrate that skeletal unloading causes an impairment of insulin effect, but not IGF-I effect, on bone metabolism in femoral tissues.

Alkaline Phosphatase↗
PubMed1994

Change in growth kinetics of hybridoma cells entrapped in collagen gel affected by alkaline supply.

The growth yields for glucose and glutamine of murine hybridoma cells entrapped in collagen gel particles were examined during the growth phase. The immobilized hybridoma cells were cultivated in a fluidized bed fermenter where the medium was circulating to supply oxygen separately. Procedures to supply an alkaline solution for adjusting the pH level strongly affected the growth yields. A direct supply of the alkaline solution to the cultivation system reduced both the growth yields for glucose and glutamine, probably due to a local increase in pH level. On the other hand, when fresh medium in which the pH was adjusted to around 8.5 was added to the cultivation system, the growth yields were unchanged even at the same pH level as when direct alkaline supply was used. These results suggest that an indirect alkaline supply could be recommended to adjust the pH level when using medium-circulating-fermenters.

Animals↗
PubMed1994

Energy-transducing nicotinamide nucleotide transhydrogenase: nucleotide sequences of the genes and predicted amino acid sequences of the subunits of the enzyme from Rhodospirillum rubrum.

Based on the amino acid sequence of the N-terminus of the soluble subunit of the Rhodospirillum rubrum nicotinamide nucleotide transhydrogenase, two oligonucleotide primers were synthesized and used to amplify the corresponding DNA segment (110 base pairs) by the polymerase chain reaction. Using this PCR product as a probe, one clone with the insert of 6.4 kbp was isolated from a genomic library of R. rubrum and sequenced. This sequence contained three open reading frames, constituting the genes nntA1, nntA2, and nntB of the R. rubrum transhydrogenase operon. The polypeptides encoded by these genes were designated alpha 1, alpha 2, and beta, respectively, and are considered to be the subunits of the R. rubrum transhydrogenase. The predicted amino acid sequence of the alpha 1 subunit (384 residues; molecular weight 40276) has considerable sequence similarity to the alpha subunit of the Escherichia coli and the N-terminal 43-kDa segment of the bovine transhydrogenases. Like the latter, it has a beta alpha beta fold in the corresponding region, and the purified, soluble alpha 1 subunit cross-reacts with antibody to the bovine N-terminal 43-kDa fragment. The predicted amino acid sequence of the beta subunit of the R. rubrum transhydrogenase (464 residues; molecular weight 47808) has extensive sequence identity with the beta subunit of the E. coli and the corresponding C-terminal sequence of the bovine transhydrogenases. The chromatophores of R. rubrum contain a 48-kDa polypeptide, which cross-reacts with antibody to the C-terminal 20-kDa fragment of the bovine transhydrogenase. The predicted amino acid sequence of the alpha 2 subunit of the R. rubrum enzyme (139 residues; molecular weight 14888) has considerable sequence identity in its C-terminal half to the corresponding segments of the bovine and the alpha subunit of the E. coli transhydrogenases.

Amino Acid Sequence↗
PubMed1994

Functional deterioration of the liver by elevated inferior vena cava pressure: a proposed upper safety limit of pressure for maintaining liver viability in dogs.

OBJECTIVE: Abnormally elevated central venous pressure is considered to be an etiological factor in the onset of acute hepatic failure following modified Fontan operation. This paper hypothesises that an increase in inferior vena cava pressure (IVCP) after such an operation has adverse effects on hepatic energy status. DESIGN: Various degrees of venous hypertension were produced in 10 mongrel dogs by clamping the thoracic IVC with an active veno-venous shunt and varying its flow rate from 60-2.5 ml/min/kg. Arterial ketone body ratio (KBR), reflecting the hepatic mitochondrial redox state, was measured as an index of hepatic energy status. MEASUREMENTS AND RESULTS: The lower the flow rates of the shunt, the higher the pressures of IVC and portal vein, while systolic blood pressure was maintained above 100 mmHg. CO significantly decreased when the pump speed was less than 30 ml/min/kg. KBR showed a negative correlation to IVCP as well as a positive correlation to portal blood flow (p < 0.05). CONCLUSION: From the simple regression line obtained between IVCP and KBR, it was determined that an upper safety limit of IVCP may lie at about 27 cmH2O (20.5 mmHg), and that a IVCP of 35 cmH2O (26.6 mmHg) seems to be the critical level for maintaining liver viability.

Animals↗
PubMed1994

Effect of different magnitudes of tension force on prostaglandin E2 production by human periodontal ligament cells.

Periodontal ligament (PDL) cells are known to produce prostaglandin E2 (PGE2) in response to mechanical stress. However, the rate of PGE2 production from PDL cells in response to different magnitudes of tension forces has not been examined. This study, therefore, was undertaken to determine the effect of different magnitudes of tension forces on PGE2 production and inositol trisphosphate (IP3) levels in PDL cells in vitro. Human PDL cells were cultured on flexible-bottomed plates and placed on a Flexercell strain unit. Cells were flexed at six cycles (5-s strain, 5-s relaxation) at six steps of tension force (9, 12, 15, 18, 21, 24% increase in surface area) for 5 days. PGE2 production and IP3 levels were determined by radioimmunoassay. There was a 6- and 25-fold increase in the rate of PGE2 production by cells exposed to low (9%) and high (24%) tension forces, respectively, and these increases were tension force-dependent. Tension force also induced increases in the intracellular levels of IP3 that did not seem to be directly related to the production of PGE2. The different rates of PGE2 production by PDL cells in response to different magnitudes of mechanical stress may be of importance in PDL and alveolar bone metabolism.

Adult↗
PubMed1994

Functional changes of dextran-modified alkaline proteinase from alkalophilic Bacillus sp.

A serine alkaline proteinase (EC 3.4.21.62) from Bacillus sp. (ALPase I) was modified with the 2,4-dialdehyde derivative of clinical dextran (dialdehyde dextran). The modified preparation was purified using an ion-exchange column and gel filtration. The modified enzyme contained 75% carbohydrate by weight. The isoelectric point (pI) of ALPase I was converted from 8.2 to approximately 5.0 by this modification. The specific activity of the dextran-modified ALPase I was 56% of that of the native enzyme when milk casein was used as a substrate. It also had some superior characteristics: the thermostability of the modified enzyme at pH 10.0 was about 10-15 degrees C higher than that of control. In organic solvents such as n-hexane, benzene, and toluene, the hydrolysis reaction of the modified ALPase I for the fluorogenic substrate, succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-4-methylcoumaryl-7-am ide (Suc-Ala-Ala-Pro-Phe-MCA), was several times higher than that of the native. This modification greatly improved the stability of ALPase I against nonionic and anionic surfactants. After exposure to lauryl benzene sulfonate and sodium lauryl sulfonate the modified enzyme retained over 95 and 90% of its activity, respectively, but the native enzyme lost its activity. We conclude that modification of serine proteinases with dialdehyde-dextran might be a useful method for improving enzyme character for enzyme technology.

Amino Acid Sequence↗
PubMed1994

Cerebral ischemia alters glucose transporter kinetics across rat brain microvascular endothelium. Quantitative analysis by an in situ brain perfusion method.

The purpose of this study was to quantify the changes of blood-brain barrier glucose transporter kinetics following cerebral ischemia using an in situ brain perfusion technique. Sixty-four adult male Sprague-Dawley rats were divided into control and ischemia groups, and a four-vessel occlusion model was used to provide an ischemic insult. To obtain regional capillary permeability area products of glucose and regional perfusion fluid flow rates, the perfusion fluid was dually labeled with 2-deoxy[14C]glucose and [3H]diazepam, and the brain was perfused at a constant rate via the external carotid artery. After sampling tissues from the brain, dual scintillation counting was performed and both regional perfusion fluid flow rates and regional capillary permeability area products were calculated. We determined kinetic parameters, including Vmax, Km and Kd as described in the Michaelis-Menten equation, by the non-linear least squares method. In the ischemia group, a decrease in Vmax and an increase in Km were recognized, which mean decreases in the affinity and the number of functioning glucose transporters. These results suggest that cerebral ischemia downregulates the blood-brain barrier glucose transporters.

Animals↗
PubMed1994

Effect of parathyroid hormone and interleukin-1 alpha in osteoblastic MC3T3-E1 cells: interaction with beta-alanyl-L-histidinato zinc.

beta-Alanyl-L-histidinato zinc (AHZ), which is an activator of bone formation, has an inhibitory effect of bone resorption. Whether AHZ can inhibit the effect of parathyroid hormone (PTH) or interleukin-1 alpha (IL-1 alpha), which is a bone resorbing factor, on osteoblastic MC3T3-E1 cells was investigated. After subculture for 3 days, the cells were cultured for 48 h with peptides. Parathyroid hormone (10(-9)-10(-7) M) or IL-1 alpha (50 U/ml) caused a significant decrease in the cellular alkaline phosphatase activity and a remarkable increase of prostaglandin E2 (PGE2) production in the cells. Parathyroid hormone (10(-7) M) or IL-1 alpha (50 U/ml) did not have an appreciable effect on the protein content of the cells. beta-Alanyl-L-histidinato zinc (10(-5) M) significantly increased the cellular alkaline phosphatase activity and protein content, whereas it had no effect on PGE2 production. This increasing effect of AHZ was also seen in the presence of PTH (10(-7) M) or IL-1 alpha (50 U/ml), although the effect of PTH and IL-1 alpha to stimulate PGE2 production was not modulated by AHZ treatment. The present finding suggests that the inhibitory effect of AHZ on bone resorption is not through osteoblasts.

Alkaline Phosphatase↗
PubMed1994

Effect of zinc-chelating dipeptide on bone metabolism in weanling rats: comparison with beta-alanyl-L-histidinato zinc-related compounds.

Rats were orally administered zinc sulfate, di(histidino)-zinc (HZ), beta-alanyl-L-histidinato zinc (AHZ), and di(N-acetyl-L-alanyl-L-histidinato)-zinc (AAHZ) with the doses of 2.75 and 5.50 mg Zn/kg/day for 7 days, and 24 h later they were killed. With the dose of 2.75 mg Zn/kg/day, the administration of AHZ caused a significant increase of zinc content in the femoral diaphysis and a corresponding elevation of alkaline phosphatase activity, deoxyribonucleic acid (DNA), and calcium contents, but similar doses of zinc sulfate, HZ, and AAHZ did not have an appreciable effect on the enzyme activity and DNA content. Moreover, the dose of 5.50 mg Zn/kg/day with AHZ and AAHZ caused an appreciable increase in all zinc compounds observed, whereas zinc sulfate did not have the effect on bone DNA content and HZ had no effect on bone alkaline phosphatase activity. Of the various zinc-chelating dipeptides used, AHZ had a characteristic effect on bone metabolism in weanling rats. Thus, AHZ may have a pharmacological role.

Animals↗
PubMed1994

Effect of zinc-chelating dipeptides on osteoblastic MC3T3-E1 cells: activation of aminoacyl-tRNA synthetase.

The effect of zinc-chelating dipeptides on osteoblastic MC3T3-E1 cells was investigated. As zinc compounds, we used zinc sulfate, AHZ, di(N-acetyl-beta-alanyl-L-histidinato)zinc (AAHZ), and di(histidino)zinc (HZ). Cells were cultured for 72 h in the presence of zinc compounds (10(-8)-10(-5) M). The effect of AHZ (10(-7) and 10(-6) M) to increase protein and deoxyribonucleic acid (DNA) contents in the cells was the greatest in comparison with those of other zinc compounds. Zinc sulfate and HZ at 10(-7) M did not have an effect on the cellular protein content. AHZ (10(-6) M) had a potent effect on cell proliferation, although zinc sulfate (10(-6) M) had no effect. beta-Alanyl-L-histidine (10(-6) and 10(-5) M) did not have an appreciable effect on the cells. Those effects of AHZ (10(-6) M) on osteoblastic cells were completely abolished by the presence of cycloheximide (10(-6) M). AHZ (10(-8)-10(-5) M) directly activated [3H]leucyl-tRNA synthetase in the cell homogenate, whereas the effect of zinc sulfate was seen at 10(-6) and 10(-5) M. The present study suggests that the chemical form of zinc-chelating beta-alanyl-L-histidine (AHZ) can reveal a potent anabolic effect on osteoblastic cells, and that AHZ directly stimulates protein synthesis.

3T3 Cells↗
PubMed1994

Identification of factors mediating the decrease of alkaline phosphatase activity caused by tension-force in periodontal ligament cells.

1. We examined the factors which mediate the decrease of alkaline phosphatase (ALP) activity in human peridontal ligament (PDL) cells in response to cyclic tension-force. 2. ALP activity in human PDL cells obtained from three donors in response to cyclic tension-force (24% elongation) was 43% lower than that of the corresponding control. 3. ALP activity was decreased by the addition of conditioned medium obtained from the culture of the cells exposed to tension-force. 4. The inhibitory effect of the conditioned medium on ALP activity was partially abolished in the presence of indomethacin (10(-6) M) and IL-1 beta antibody (10 ng/well). Moreover it was almost completely abolished in the presence of both indomethacin and IL-1 beta antibody. 5. Treatment of PDL cells with exogenous PGE2 or IL-1 beta for 24 hr caused a dose-dependent decrease in ALP activity. Treatment with both PGE2 (10(-8) M) and IL-1 beta (1.25 x 10(-10) M) together decreased ALP activity by 47% compared with the non-treated control. 6. These findings suggest that ALP activity in PDL cells was decreased in response to the cyclic tension-force and that the decrease in ALP activity was mainly mediated by PGE2 and IL-1 beta produced by PDL cells in response to cyclic tension-force.

Alkaline Phosphatase↗
PubMed1994

Ultracytochemistry of glycoconjugates in pig duodenal gland.

To elucidate the ultrastructure, glycoprotein profile and site of glycosylation of glandular cells in relation to the functional polarity of the organelles, swine duodenal tissue was embedded in glycol methacrylate and subsequently stained for periodic acid thiocarbohydrazide silver proteinate (PA-TCH-SP), high iron diamine (HID)-TCH-SP, low iron diamine (LID)-TCH-SP, ninhydrin T-TCH-SP and five peroxidase-labeled lectins. The secretory granules in the duodenal gland cells were electron lucent with a 200 nm to 500 nm electrondense core. Glycoconjugates were confined to the secretory granules and elements of the Golgi complex. Protein activity was located only in the electron dense core. Achivementestic staining pattern for Concanavalin A (Con A), peanut agglutinin (PNA), Ulex europaeus agglutinin-I (UEA-I), wheat germ agglutinin (WGA) and soybean agglutinin (SBA) was observed in stained secretory granules and the Golgi apparatus. A few cis cisternae were stained with SBA and UEA-I. Trans cisternae were stained with WGA and PNA. Con A reacted with seromucous granules and rough endoplasmic reticulum. These observational findings suggest that these are seromucous cells. The Golgi apparatus is the site of glycosylation and can be divided into two distinct compartments.

Animals↗
PubMed1994

Natriuretic peptide-potentiating actions of neutral endopeptidase inhibition in rats with experimental heart failure.

We developed a rat model of heart failure induced by myocardial infarction (MI) which preserves responsiveness to exogenously administered natriuretic peptide, and investigated the potentiating action of neutral endopeptidase (NEP) inhibition on the renal response to endogenous natriuretic peptide in MI rats, comparing with that in the established cardiac-failing model with arterio-venous fistula (AVF). The endogenous plasma concentration of alpha-rat atrial natriuretic peptide (alpha-rANP) in the MI rat was 6.4-fold higher than that in the normal rat, and intravenous infusion of phosphoramidon (165 nmol/min/kg), an NEP inhibitor, induced larger increases in circulating alpha-rANP levels and natriuresis in MI rats than in normal controls. The maximal natriuretic effect of phosphoramidon (165 nmol/min/kg) was equal to that of exogenously administered alpha-rANP (100 pmol/min/kg) in MI rats, whereas plasma alpha-rANP concentration under NEP inhibition was much lower than that after administration of alpha-rANP. The endogenous alpha-rANP levels in AVF rats were as high as those in MI rats. However, the natriuretic effect of phosphoramidon was less in AVF rats than in MI rats, which was consistent with the decreased natriuretic activity observed with administration of exogenous to alpha-rANP in the AVF rat. These results indicate that the natriuretic effect of NEP inhibition is dependent on elevated endogenous alpha-rANP levels in cardiac-failing rats, but cannot be accounted for simply in terms of the increase in circulating alpha-rANP levels. Endogenous natriuretic peptide-mediated natriuresis under NEP inhibition also appears to correlate with the responsiveness to the exogenously administered peptide.

Animals↗
PubMed1994

Clearance mechanisms of atrial and brain natriuretic peptides in rats.

To assess clearance mechanisms of atrial and brain natriuretic peptides in the circulation, we examined the effects of a neutral endopeptidase (NEP) inhibitor and a clearance receptor ligand on plasma concentrations of the peptides in normal rats. Plasma concentrations of endogenous alpha-rat atrial natriuretic peptide (alpha-rANP) were not significantly elevated by intravenous infusion of a NEP inhibitor, phosphoramidon, but were elevated threefold by intravenous infusion of a clearance receptor ligand, des(Gln18-Gly22)-rANP(4-23)-NH2 [C-ANF(4-23)]. On the other hand, the clearance of alpha-rANP given intravenously at the pharmacological dose, 600 pmol/min/kg for 2 min, was decreased to one-third by the administration of phosphoramidon, although the administration of C-ANF(4-23) did not significantly decrease the clearance. The clearance of rat brain natriuretic peptide (rBNP) given at 600 pmol/min/kg for 2 min was approximately 38% lower than that of alpha-rANP. The effect of phosphoramidon on the clearance of rBNP was not significant and was similar to that of C-ANF(4-23). These results suggest that clearance receptor is involved in the clearance of the physiological levels of alpha-rANP and that NEP plays a major role in the clearance of a pharmacological dose of alpha-rANP, at which clearance receptors are thought to be saturated, and also indicate a pharmacokinetic difference between alpha-rANP and rBNP.

Amino Acid Sequence↗
PubMed1994

HLA-B*3501-peptide interactions: role of anchor residues of peptides in their binding to HLA-B*3501 molecules.

Two HLA-B*3501 binding self-peptides, LPFDFTPGY (37F) and LPGPKFLQY (28H), were isolated from HLA-B*3501 molecules expressed by cultured human B lymphoid cells. Both sequences were consistent with previously reported motifs of HLA-B*3501 binding peptides which carry proline at position 2 and tyrosine at position 9 as anchor residues. Direct binding of these peptides to HLA-B*3501 molecules was quantitated by flow cytometry analysis of RMA-S cells. transfected with the HLA-B*3501 gene (RMA-S-B*3501). Both 37F and 28H peptides bound effectively to HLA-B*3501 molecules. Substitution of amino acids at position 2 and/or 9 of HLA-B*3501 binding peptides markedly reduced their binding to HLA-B*3501 molecules. These results indicate that two anchor residues, proline at position 2 and tyrosine at position 9 are critical in binding of peptides to HLA-B*3501 molecules. Insertion of up to four glycine residues at position 8 of the peptide 37F did not affect its binding affinity to HLA*3501 molecules. These results indicate that long peptides can effectively bind to HLA class I molecules provided that anchor residues are conserved.

Amino Acid Sequence↗