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Biomedical subjects

K Shirai

Publications and source records attributed to K Shirai.

At least 73 records · Page 4Linked to original sources

Influence of polymerization initiator for base monomer on microwave curing of composite resin inlays.

Microwave polymerization was used to make composite resin inlays and the effect examined of the concentration of polymerization initiator for the base monomer. The monomers used were 2,2-bis [4-(3-methacriloxy-2-hydroxypropoxy) phenyl] propane (Bis-GMA) and triethyleneglycol dimethacrylate (TEGDMA). Bis-GMA and TEGDMA were mixed in a ratio of 6:4 by weight and were separated into five groups. To each group was added benzoyl peroxide (BPO) in the ratios of 0.1, 0.3, 0.5, 0.7 and 0.9 wt% as the polymerization initiator. These were used as the base monomers. The results showed that the degree of conversion of the cured sample increased with increasing concentration of BPO from 0.1 to 0.5 wt%, however there was no significant difference at 0.5, 0.7 and 0.9 wt% (P> 0.01). Compression strength, diametral tensile strength and the Knoop hardness showed a similar tendency as the degree of conversion. No significant difference was recorded in the Knoop hardness between the top and the bottom surfaces (P> 0.01), which suggested a uniform polymerization in the cured sample. Thus, microwave polymerization would be an efficacious method for making resin inlays with the addition of BPO to the base monomer (Bis-GMA:TEGDMA, 6:4). The maximum conversion was found at a concentration of 0.5 wt%.

Benzoyl Peroxide↗

Immunolocalization of transcription factor AP1 in human ocular surface epithelia.

PURPOSE: The present study examined whether normal human ocular surfcae epithelia express AP1 components. Changes in expression patterns of these components in a case of ocular surface epithelial dysplasia was also evaluated before and after topical mitomycin C treatment. METHODS: Specimens of normal corneas (n = 2) and conjunctiva (n = 4) were obtained from 4 patients during cataract surgery or post mortem, while specimens of dysplastic epithelial tissue from the limbus were obtained from one patient. Specimens were immunohistochemically studied using antibodies against components of AP1. RESULTS: The normal corneal epithelium showed no staining with antibodies against c-Fos, Fra-2, FosB, c-Jun or JunB, whereas the limbal and bulbar conjunctival epithelia were positive for c-Fos, Fra-2, and c-Jun. Anti-FosB and -JunB antibodies reacted weakly with the conjunctival epithelium. JunD was absent in normal corneal and conjunctival epithelia. The dsyplastic epithelium showed positive labelling for c-Fos, Fra-2, c-Jun, and JunD throughout its thickness. Fra-1 was present in all specimens of epithelia examined. The dysplastic epithelium treated with mitomycin C was not labeled by anti-c-Fos or -Fra-2 antibody. CONCLUSION: Individual AP1 components show specific expression patterns in normal ocular surface epithelia and a case of dysplastic epithelium before and after topical MMC treatment, implying that these factors may play important roles in modulating epithelial cell function, e.g., proliferation and differentiation.

Adult↗

Modification of lipoprotein lipase catalytic activity by sialic acids.

The role of sialic acid linked with lipoprotein lipase (LPL) in its catalytic activity was studied. When LPL was treated with sialidase, the molecular weight decreased by 2000. The sialidase-treated LPL showed unchanged hydrolyzing activity for tributyrin, a water-soluble substrate of esterase, compared with the untreated LPL. The sialidase-treated LPL also showed similar hydrolyzing activity for triolein emulsified with Triton X-100, phosphatidylcholine and phosphatidylethanolamine, whereas it showed significantly increased hydrolyzing activity for triolein emulsified with phosphatidylserine and cardiolipin (152% and 183%, compared with untreated LPL, respectively). In addition, the sialidase-treated LPL showed significantly increased hydrolyzing activity against triolein incorporated into very low-density lipoproteins and chylomicrons (151% and 186%, compared with the untreated LPL, respectively). These results suggest that the loss of sialic acids does not modify the function of the catalytic site of LPL, but facilitates the interaction of the enzyme with the interface of the surface of substrate lipoproteins.

Animals↗

A missense mutation of the nitric oxide synthase (eNOS) gene (Glu298Asp) in five patients with coronary artery disease--case reports.

The authors report five patients with a missense mutation of the endothelial nitric oxide synthase (eNOS) gene (Glu298Asp) who have angiographically proven coronary artery disease (CAD). They compare their clinical findings and coronary arteriographic characteristics. They conclude that these case reports show that this mutation is not solely responsible for development of CAD. Diabetes mellitus, smoking, and hyperlipidemia are other risk factors.

Aged↗

Levels of soluble cell adhesion molecules in patients with angiographically defined coronary atherosclerosis.

Adhesion molecules on the endothelial cell membrane play an important role in the pathogenesis of atherosclerosis. Levels of soluble forms of cell adhesion molecules are reportedly elevated in patients with peripheral artery vessel disease and in patients with an atherosclerotic aorta. The present study investigated the association of serum levels of soluble vascular cell adhesion molecule 1 (sVCAM-1), soluble intercellular adhesion molecule 1 (sICAM-1), and soluble P-selectin (sP-selectin) with coronary heart disease (CHD) and the extent of coronary atherosclerosis, and examined the influence of serum levels of lipids, lipoproteins and apolipoproteins (apo) in subjects with (n=52, M/F:43/9) and without (controls, n=40, M/F:25/15) angiographically proven coronary atherosclerosis. After controlling for age and gender, levels of sVCAM-1 (least squares mean +/- std error: 565+/-36 ng/ml vs 540+/-41 ng/ml, ns), sICAM-1 (261+/-17ng/ml vs 247+/-19ng/ml, ns), and sP-selectin (142+/-8ng/ml vs 149+/-10 ng/ml, ns) in patients with coronary atherosclerosis were not different from those in controls, as assessed by an analysis of covariance. After also adjusting for body mass index, hypertension, diabetes mellitus, and smoking by a multiple logistic function analysis, the association of sVCAM-1, sICAM-1, and sP-selectin with CHD was still not significant. Levels of sVCAM-1, sICAM-1, and sP-selectin were also not related to the extent of coronary atherosclerosis as judged by the number of stenosed vessels. However, inverse (p<0.05) relationships were observed between sVCAMs and serum levels of HDL3-cholesterol, apo A-II, and lipoprotein containing apo A-I and A-II, between sICAMs and levels of apo A-II and Lp A-I/A-II (Lp A-I/A-II), and between sP-selectin and lipoprotein containing only apo A-I. In conclusion, serum levels of soluble VCAM-1, ICAM-1, and P-selectin were not related to CHD or the extent of coronary atherosclerosis, but were inversely related to serum levels of high-density lipoprotein-related lipoproteins.

Apolipoproteins↗

Associations among serum lipoprotein(a) levels, apolipoprotein(a) phenotypes, and myocardial infarction in patients with extremely low and high levels of serum lipoprotein(a).

A high serum lipoprotein(a) [Lp(a)] level, which is genetically determined by apolipoprotein(a) [apo(a)] size polymorphism, is an independent risk factor for coronary atherosclerosis. However, the associations among Lp(a) levels, apo(a) phenotypes, and myocardial infarction (MI) have not been studied. Patients with MI (cases, n = 101, M/F: 86/15, age: 62+/-10y) and control subjects (n = 92, M/F: 53/39, age: 58+/-14y) were classified into quintile groups (Groups I to V) according to Lp(a) levels. Apo(a) isoform phenotyping was performed by a sensitive, high-resolution technique using sodium dodecyl sulfate-agarose/gradient polyacrylamide gel electrophoresis (3-6%), which identified 26 different apo(a) phenotypes, including a null type. Groups with higher Lp(a) levels (Groups II, III, and V) had higher percentages of MI patients than that with the lowest Lp(a) levels (Group I) (54%, 56%, or 75% vs. 32%, p<0.05). Groups with different Lp(a) levels had different frequency distributions of apo(a) isoprotein phenotypes: Groups II, III, IV, and V, which had increasing Lp(a) levels, had increasingly higher percentages of smaller isoforms (A1-A4, A5-A9) and decreasingly lower percentages of large isoforms (A10-A20, A21-A25) compared to Group I. An apparent inverse relationship existed between Lp(a) and the apo(a) phenotype. Subjects with the highest Lp(a) levels (Group V) had significantly (p<0.05) higher serum levels of total cholesterol, apo B, and Lp(a). Patients with MI and the controls had different distributions of apo(a) phenotypes: i.e., more small isoforms and more large size isoforms, respectively (A1-A4/A5-A9/A10-A20/A21-A25: 35.7%/27.7%/20.8%/15.8% and 22.8%/23.9%/29.4%/23.9%, respectively). Lp(a) (parameter estimate +/- standard error: 0.70+/-0.20, Wald chi2 = 12.4, p = 0.0004), apo(a) phenotype (-0.43+/-0.15, Wald chi2 = 8.17, p = 0.004), High-density lipoprotein-cholesterol, apo A-I, and apo B were significantly associated with MI after adjusting for age, gender, and conventional risk factors, as assessed by a univariate logistic regression analysis. The association between Lp(a) and MI was independent of the apo(a) phenotype, but the association between the apo(a) phenotype and MI was not independent of Lp(a), as assessed by a multivariate logistic regression analysis. This association was not influenced by other MI- or Lp(a)-related lipid variables. These results suggest that apo(a) phenotype contributes to, but does not completely explain, the increased Lp(a) levels in MI. A stepwise logistic regression analysis with and without Lp(a) in the model identified Lp(a) and the apo(a) phenotype as significant predictors for MI, respectively.

Apolipoproteins A↗

Insulin resistance and angiographical characteristics of coronary atherosclerosis.

Insulin resistance (IR) is frequently observed in patients with coronary heart disease (CHD). The relationship between IR and the angiographical characteristics of coronary atherosclerosis were investigated in 66 patients with coronary artery lesions. Insulin resistance was assessed by a 75-g oral glucose tolerance test and homeostasis model assessment (HOMA). The angiographical characteristics of coronary atherosclerosis (i.e., the severity of CHD) were defined by both Gensini's score (GS) (a higher degree of coronary artery stenosis or a proximal lesion was assigned a higher score than a distal lesion) and the number of significantly stenosed vessels. When GS was examined as a categorical variable classified by tertile values (Group A, n = 22: 1< or =GS< or =14; Group B, n = 22: 15< or =GS< or =32; and Group C, n = 22: 33< or =GS), patients with a high GS (Group C) had significantly (p<0.05) higher values of fasting plasma insulin, insulin response, and HOMA IR than patients with a low GS (Group A) (12.6+/-1.2 microU/ml vs. 6.9+/-1.2 microU/ml, 122.2+/-11.9 microU ml(-1) h(-1) vs. 72.9+/-12.9 microU ml(-1) h(-1), and 2.9+/-0.3 vs. 1.5+/-0.3, respectively). The values in Group B patients (9.4+/-1.2,microU/mI, 108.5+/-12.5 microU ml(-1) h(-1), and 2.1+/-0.3, respectively) were intermediate between those in Groups A and C. The area of insulin/area of glucose ratio was significantly (p<0.05) higher in Groups B and C than in Group A (0.54+/-0.06 microU/mg, 0.54+/-0.06 microU/mg, and 0.32+/-0.06 microU/mg, respectively). However, no significant differences were observed in variables of glucose tolerance, serum lipid, lipoproteins, fibrinogen, uric acid, and blood pressure among the 3 groups. Significant (p<0.05) positive associations were found between GS, the number of diseased coronary arteries, and fasting immunoreactive insulin, insulin response, the area of insulin/area of glucose ratio and HOMA IR by logistic regression analysis. After adjusting for the number of diseased coronary arteries, the association between GS and IR was not significant, suggesting that IR contributed to the severity of coronary atherosclerosis but not to the distribution of lesions. In conclusion, IR was associated with the severity of CHD as measured by both Gensini's score and the number of diseased coronary arteries, and increased the risk of CHD regardless of the location of the lesions.

Blood Pressure↗

Cloning of a thermostable ascorbate oxidase gene from Acremonium sp. HI-25 and modification of the azide sensitivity of the enzyme by site-directed mutagenesis.

A gene encoding a thermostable ascorbate oxidase (ASOM) was cloned from Acremonium sp. HI-25 and sequenced. The gene comprised 1709 bp and was interrupted by a single intron of 57 bp. ASOM consisted of 551 amino acids including a signal peptide with a molecular mass of 61200, and contained four histidine-rich regions with high sequence homology to the corresponding regions of other multicopper oxidases. The ASOM gene was expressed in Aspergillus nidulans under the Aspergillus oryzae Taka-amylase A gene promoter. The recombinant enzyme (An-ASOM) exhibited almost the same enzymatic properties as ASOM. The ASOM gene was mutated by site-directed mutagenesis with reference to the amino acid sequences of plant enzymes to generate enzymes with altered azide sensitivity. Site-directed mutagenesis at the trinuclear active copper site resulted in an increase in azide resistance; the Ala465Leu and Phe463Trp/Ala465Leu mutants exhibited approximately 10 and 20% increases in azide resistance, respectively.

Acremonium↗

Subretinal administration of tissue-type plasminogen activator to speed the drainage of subretinal hemorrhage.

BACKGROUND: Bleeding into the subretinal space in the vicinity of the macula is associated with age-related macular degeneration or retinal arterial macroaneurysm. The prognosis for restoration of vision is poor in the presence of blood clots. METHODS: Using a simple device composed of three disposable syringes we injected tissue-type plasminogen activator (tPA) into the subretinal space during conventional vitrectomy in six patients to assist the draining of subretinal clots. RESULTS: Four of six patients recovered their visual acuity postoperatively, while visual acuity in the other patients was stabilized. CONCLUSION: Early drainage of subretinal hemorrhage assisted by the introduction of tPA into the subretinal space led to uncomplicated surgery and favorable postoperative results.

Aged↗

Immunolocalization of proto-oncogene products in keratocytes after epithelial ablation, alkali burn and penetrating injury of the cornea in rats.

BACKGROUND: We examined the immunolocalization of proto-oncogene products, including c-Fos and c-Jun, in the rat cornea during epithelial and stromal wound healing after simple epithelial ablation, penetrating injury or alkali burn. METHODS: Eighty-four male Wistar rats were divided into three groups and subjected to treatments as follows: (a) ablation of central corneal epithelium leaving basement membrane intact, (b) alkali burn in the central cornea with 1 N NaOH and (c) penetrating injury at the central cornea. The affected eyes were then enucleated after various intervals of healing. The frozen sections were immunohistochemically stained with the antibodies against proto-oncogene products. RESULTS: c-Fos- and c-Jun-immunoreactive cells were detected in the epithelium around the epithelial defect from 60 to 120 min after these treatments. The distribution of these cells were varied in these three types of injury. The immunoreactivities for these proteins were also detected in keratocytes after epithelial ablation. In the corneas with alkali burn, the immunoreactivities were detected in the keratocytes in the whole corneal stroma, and these immunoreactions were stronger than those observed in simple epithelial ablation. In the corneas with penetrating injury, such immunoreaction was seen only in keratocytes around the wound. CONCLUSION: These findings indicate that activator protein 1-mediated transcriptional activation for epithelial migration is initiated in the early phase after each injury, and that stromal keratocytes are also transcriptionally activated not only by alkali burn or penetrating injury but also by simple epithelial ablation in which basement membrane was not affected.

Animals↗

An atherogenic midband lipoprotein: a risk factor for coronary artery disease in diabetes mellitus with hyperlipidemia.

On polyacrylamide gel (PAG) disc electrophoresis of serum lipoproteins, the band(s) which migrates between pre beta- and beta-lipoproteins was more frequently observed in hyperlipidemics with diabetes mellitus (73%), than in those without diabetes mellitus (37%) (P < 0.01). Those bands were seen at three positions between pre beta- and beta-lipoproteins. A higher incidence of coronary artery disease (CAD) was observed in patients with midband as a shoulder of beta-lipoproteins (44%), than in those without midband (11%) (P < 0.05) after the matching of other risk factors. These results suggest that midband as a shoulder of beta-lipoproteins may be a risk factor for CAD in diabetes mellitus with hyperlipidemia.

Coronary Disease↗

Electrocardiographic changes related to parasympathetic tone during right coronary angioplasty.

The role of basal parasympathetic tone in reducing the heart rate and causing electrocardiographic (ECG) changes during right coronary angioplasty was evaluated in 19 patients with angina pectoris. To measure parasympathetic tone, time and frequency domain parameters of heart rate variability were assessed using 24 h of ambulatory ECG data recorded before angioplasty. There was an age-dependent reduction in heart rate. However, no parameters of heart rate variability were responsible for the changes in heart rate. In contrast, the degree of ST segment elevation during angioplasty correlated significantly with the high-frequency component of heart rate variability. Therefore, the increased parasympathetic tone, as assessed by spectral analysis, may augment the early ST segment elevation induced by right coronary occlusion.

Aged↗

Parasympathetic tone affects electrocardiographic preconditioning during right coronary angioplasty.

We investigated whether parasympathetic tone may affect ischemic preconditioning in terms of ST segment alteration during coronary angioplasty. Coronary balloon angioplasty was performed in 19 patients with angina and isolated right coronary artery disease. Parasympathetic tone was assessed by frequency domain parameters of heart rate variability using 24-hour electrocardiographic recordings obtained before angioplasty. Logistic regression analysis showed the high frequency component to be an independent predictor of the change in ST elevation induced by the second balloon inflation. An increase in parasympathetic tone was related to ischemic preconditioning with respect to the electrocardiographic alterations observed in these patients.

Aged↗

Primary pulmonary artery sarcoma.

A 66-year-old woman with a history of mitral valve replacement with a Starr-Edwards ball valve 25 years ago was treated for refractory heart failure but died of right heart failure. At autopsy, primary pulmonary artery sarcoma was found in the right ventricular outflow tract, main pulmonary trunk, and bilateral pulmonary artery, and had invaded the aortic arch. The pathohistologic diagnosis was osteosarcoma. Echocardiography, chest computed tomography and right ventriculography performed 1 year before death did not reveal the presence of a tumor in the pulmonary artery. The history of this patient shows that primary pulmonary artery sarcoma grows rapidly, with, in this case, the patient dying within 1 year of its appearance.

Aged↗

Molecular cloning of bottle-nosed dolphin (Tursiops truncatus) MHC class I cDNA.

Using polymerase chain reaction, bottle-nosed dolphin (Tursiops truncatus) major histocompatibility complex (DoLA) class I alpha chain cDNA was cloned and sequenced. Predicted amino acid sequences of DoLA class I alpha chain have cystein residues for intradomain disulfide bond formation and N-linked glycosylation sites, suggesting that DoLA class I alpha chain molecules construct alpha 1, alpha 2, and alpha 3 domain structures. Similarity of DoLA class I alpha chain cDNA to land mammal MHC class I alpha chain cDNA was 90.4% in cattle, 90.2% in horse, 89.4% in sheep, and 87.8% in human. This investigation suggests that DoLA is closely related to land mammals.

Amino Acid Sequence↗

A monoclonal antibody against dolphin lymphocytes (6E9) which recognizes bovine MHC class II antigens.

A monoclonal antibody, 6E9 established from mice injected with dolphin peripheral blood lymphocytes (PBLs) was characterized. In addition to its reactivity against 89.4% of dolphin PBLs, 6E9 reacted with 33.1% of bovine PBLs of which 22% were CD5+, 11.1% were CD5-. 6E9 recognized a 34 kD protein on the surface of dolphin and bovine PBLs. Analysis of the protein's N-terminal amino acid sequence indicated that 6E9 recognizes bovine major histocompatibility complex (MHC) class II antigens. These results suggested that 6E9 recognized MHC class II antigens on bovine PBLs. As we have already produced an anti-dolphin MHC class I monoclonal antibody, analysis of immune system using these monoclonal antibodies will advance our understanding of the evolution of the mammalian immune system.

Animals↗

Long-term effect of low-density lipoprotein apheresis in a patient with heterozygous familial hypercholesterolemia: follow-up study using coronary angiography.

A 62-year-old man with old myocardial infarction and familial hypercholesterolemia was treated by both probucol and low-density lipoprotein (LDL) apheresis. Coronary angiography was performed before and after 3.5 years of LDL apheresis treatment, and no new lesion or progression of coronary atherosclerosis was observed. LDL apheresis drastically reduced the serum total cholesterol. However, it is still unclear whether LDL apheresis effectively prevented the recurrence of cardiac events and the progression of coronary atherosclerosis in this patient.

Anticholesteremic Agents↗