Biomedical subjects

Y Nakashima

Publications and source records attributed to Y Nakashima.

At least 109 records · Page 6Linked to original sources

PubMed2000

Fas-Fas ligand system in the peripheral blood of patients with renal diseases.

To clarify the role of the Fas-Fas ligand (FasL) system in the peripheral blood from patients with various renal diseases, the Fas and FasL expression on mononuclear cells (MNCs) and serum levels of soluble Fas (sFas) and soluble FasL were investigated. Patients were selected from those with various types of glomerular diseases showing various degrees of renal function. Fas expression on MNCs was analyzed by a FACScan, sFas and soluble FasL were measured with an ELISA kit, and FasL expression on MNCs was counted using a FACScan after a bioassay. Fas-positive MNCs and sFas increased with statistical significance concomitantly with deterioration in renal function. Moreover, there was a significant correlation between them. sFas- and FasL-positive MNCs were significantly correlated with proteinuria. However, the Fas expression percentage on MNCs and/or serum levels of sFas did not correlate with the number of TUNEL-positive cells in the glomeruli. Also, there was no disease specificity in the activation of Fas. These results indicate that Fas expression on MNCs is activated in accordance with the deterioration in renal function without disease specificity, corresponding to the elevation of serum sFas levels to protect against Fas-mediated apoptosis.

Adult↗

PubMed2000

Cloning of the gene of beta-N-acetylglucosaminidase from Lactobacillus casei ATCC 27092 and characterization of the enzyme expressed in Escherichia coli.

The gene encoding beta-N-acetylglucosaminidase (GlcNAcase) of Lactobacillus casei ATCC 27092 was cloned and expressed in Escherichia coli. The gene consisted of 1581 nucleotides, and had a promoter, Shine-Dalgarno, and rho-independent type transcription termination sequences typical in bacteria. The protein deduced from the sequence consisted of 526 amino acids, and had a putative signal peptide of 14 amino acids and 5 possible asparagine-linked glycosylation sites. A conserved sequence was homologous to the 12 other hexosaminidases from different origins. The recombinant GlcNAcase (r-GlcNAcase) purified from the transformed E. coli had a MW of 39 kDa and lacked oligosaccharide chains. The isoelectric point and the optimum pH for the activity of r-GlcNAcase were similar to those of original GlcNAcases (o-GlcNAcase). However, the thermal stability was lower, and sensitivity to Cd2+, Fe2+, Cu2+ and sodium dodecyl sulfate (SDS) was higher than those of o-GlcNAcases, suggesting that the oligosaccharide moieties of the enzyme contribute to their stability. The Km value for p-nitrophenyl-N-acetyl-beta-1,4-D-glucosamine (PNP-GlcNAc) of r-GlcNAcase (6.4 microM) implied that the affinity of r-GlcNAcase for the substrate was 200-fold higher than that of the original ones.

Acetylglucosaminidase↗

PubMed2000

The effect of the atrioventricular interval during atrioventricular sequential pacing on the hemodynamics in dynamic obstruction of the left ventricular outflow tract in dogs.

The present study was performed to clarify the mechanism involved in the reduction of the pressure gradient in the left ventricular outflow tract of patients with hypertrophic obstructive cardiomyopathy when treated with atrioventricular (AV) sequential pacing. The effect of AV sequential pacing with variable AV intervals on the hemodynamics and dyssynchronous wall motion was experimentally studied using echocardiography in the dynamic obstruction of the left ventricular outflow tract created by dobutamine infusion in 17 dogs. The pressure gradient of the left ventricular outflow tract decreased with shortening of the AV interval during AV sequential pacing. Also, the dyssynchrony time, defined as the difference in the time between the intraventricular septum and posterior wall during the systolic phase recorded with M-mode echocardiography, increased with shortening of the AV interval during AV sequential pacing. However, very short AV intervals produced a significant decrease in the aortic pressure and increase in the pulmonary capillary wedge pressure. The dyssynchrony time showed a positive liner correlation with the percentage reduction in the pressure gradient of the left ventricular outflow tract (R=0.794, p<0.0001). It was concluded that dyssynchronous wall motion in the left ventricle was produced by pacing from the right ventricular apex and resulted in a reduction in the pressure gradient of the left ventricular outflow tract. Optimization of the AV interval during AV sequential pacing may play an important role in improving the hemodynamics in dynamic obstruction of the left ventricular outflow tract.

Animals↗

PubMed2000

Successful treatment of malignant neurocardiogenic syncope with repeated tilt training program.

Recent reports have shown that repeated tilt-table testing or tilt training is a very effective therapy for the treatment of neurocardiogenic syncope induced by head-up tilt testing. The present patient experienced repeated syncopal or presyncopal attacks and had shown prolonged asystole on an electrocardiogram during syncope. The presyncope could be reproducibly induced by head-up tilt testing. Oral propranolol and/or disopyramide therapies failed to prevent his symptoms. Tilt training (2 sessions/day) was repeated every day for 4 weeks at home, and then head-up tilt testing was performed again. The syncope or presyncope was not induced by head-up tilt testing. The patient has continued with this training and has had no symptoms during the follow-up period of 1 year.

Electrocardiography↗

PubMed2000

Effect of a single oral dose of pilsicainide on pacing thresholds in pacemaker patients with and without paroxysmal atrial fibrillation.

A single oral dose of pilsicainide, a class 1c antiarrhythmic drug, is effective in terminating acute-onset atrial fibrillation (AF), but its effect on pacing thresholds in pacemaker patients is unknown. The present study measured atrial and ventricular pacing thresholds after a single oral dose of pilsicainide in patients with and without AF. Twelve patients with dual-chamber pacemakers were evaluated. Pacing thresholds as well as plasma pilsicainide concentration were measured prior to and then at 30, 60, 90, 120 and 180 min and 24h following a single oral dose of pilsicainide (150 mg). Six patients had paroxysmal AF and the remaining 6 did not. Pacing thresholds increased significantly (134+/-8%) in the atrium (p<0.05) and in the ventricle (155+/-11%; p<0.001) following pilsicainide administration in all 12 patients. Plasma concentrations of pilsicainide showed a positive liner correlation with pacing thresholds (R=0.62, p<0.0001 in the atrium; R=0.74, p<0.0001 in the ventricle). Atrial pacing thresholds in the patients with AF showed a significant increase at 90, 120 and 180 min compared with the patients without AF (p<0.05). There was no significant difference in either the ventricular pacing threshold or the plasma pilsicainide concentration in the patients with and without AF. It was concluded that a single oral dose of pilsicainide increases the pacing thresholds in both the atrium and ventricle in a selected group of pacemaker-implanted patients; that is, those who are aged and with AF. Thus, careful attention should be paid to pacemaker-dependent patients, particularly those with paroxysmal AF, when administering pilsicainide.

Administration, Oral↗

PubMed2000

Clinical surveillance of a thin bipolar pacing lead.

A total number of 415 co-radial, bipolar pacing leads (189 atrial leads; 226 ventricular leads) were implanted in 228 patients between November 1994 and July 1999. Mean pacing thresholds at the implantation were normal at 0.6 V in the atrium and at 0.4 V in the ventricle with a pulse duration of 0.4-0.5 ms. Lead impedance was relatively low (337-447 ohms for atrial leads; 369-459 ohms for ventricular leads) at the implantation and during the follow-up periods. No definite failure in lead materials was observed in either atrial or ventricular leads (mean follow-up of 19.7 and 19.2 months, respectively: up to 52.9 months for both leads). Predicted clinical surveillance up to 10 years calculated statistically showed that the upper 95% confidence limit was a constant of 100%. The lower 95% confidence limits at 5, 7, and 10 years were estimated to be 98.0%, 97.2%, and 96.0%, respectively. From the present study, the ThinLine lead is reliable for both sensing and pacing thresholds, and has excellent predicted lead longevity. Nevertheless, further observation is required regarding cost performance, such as early replacement of the pacemaker generator, because of the lower pacing lead impedance.

Aged↗

PubMed2000

Treatment of hyperlipidemia.

In the treatment of hyperlipidemia, when to begin and end therapy is important. In recent years, potent anti-hyperlipidemia drugs have been widely used, and the results of many intervention trials have shown that combinations of diet, exercise and drug therapies are effective for the primary and secondary prevention of coronary heart disease. The present paper summarizes these trials; introduces the therapy guidelines for adult hyperlipidemia established by Japan Atherosclerosis Society in 1997; and discusses the drugs for hyperlipidemia.

Clinical Trials as Topic↗

PubMed2000

Molecular cloning of cDNA encoding Xenopus laevis deoxyribonuclease I.

A 1200-bp cDNA encoding Xenopus laevis deoxyribonuclease I (X. laevis DNase I) was constructed from the total RNA of a X. laevis pancreas using a rapid amplification of cDNA ends method. When the cDNA was transiently transfected into COS-7 cells, the recombinant polypeptide exhibited similar enzymological properties to those of the native pancreatic DNase I. The recombinant enzyme was considerably more labile than most other vertebrate DNase I enzymes. The X. laevis DNase I polypeptide was larger than any other known vertebrate DNase I, containing a unique Cys-rich stretch of 68 or 70 amino acid residues at the carboxyl terminus, and it had less well conserved binding sites for the Ca2+, G-actin and DNA, and two DNase I signature motifs. These alterations might account for its heat instability.

Amino Acid Sequence↗

PubMed2000

Clinicopathologic study on hepatocellular carcinoma negative for hepatitis B surface antigen and antibody to hepatitis C virus.

Among 326 consecutively resected cases of hepatocellular carcinoma (HCC), we studied clinicopathologic features of 13 cases (NBNC-HCC) negative for hepatitis B surface antigen (HBsAg), antibody to HBsAg (HBsAb), antibody to hepatitis B core antigen (HBcAb), hepatitis B envelope antigen (HBeAg), antibody to HBeAg (HBeAb), and antibody to hepatitis C virus (HCVAb). Forty-four HCC cases positive only for HBsAg (B-HCC) and 232 cases positive only for HCVAb (C-HCC) were studied as controls. Clinically, NBNC-HCC cases showed good liver function reserve. None of NBNC-HCC cases had periodic medical check ups and HCC was detected in 12 cases (92.3%) incidentally. Pathologically, the mean tumor diameter was significantly larger and the histologic grade was less differentiated in NBNC-HCC cases than in B-HCC and C-HCC cases. In the background liver, liver cirrhosis was associated in 15.4% of NBNC-HCC cases, 50.0% of B-HCC and 38.2% of C-HCC cases. The degree of inflammation and fibrosis was less in NBNC-HCC cases, and two cases (15.4%) had almost normal liver histology. In NBNC-HCC cases, synchronous and metachronous multicentric occurrence was not observed. In B-HCC cases, synchronous multicentric occurrence was found in 1 case (20.0%), but no metachronous multicentric occurrence. In C-HCC cases, synchronous and metachronous multicentric occurrence was found in 13 (43.3%) and 8 (30.8%) cases, respectively. However, the cumulative recurrence-free rate was not significantly different among the three groups. Accordingly, it was suggested that better prognosis could be expected in NBNC-HCC cases compared with B- and C-HCC cases, if the cancer could be detected in the early stage.

Aged↗

PubMed2000

Expression levels of thymidine phosphorylase and dihydropyrimidine dehydrogenase in various human tumor tissues.

Thymidine phosphorylase (dThdPase) is the rate-limiting enzyme that metabolizes 5'-deoxy-5-fluorouridine (5'-dFUrd, doxifluridine), an intermediate metabolite of capecitabine, to the active drug 5-fluorouracil (5-FUra), while dihydropyrimidine dehydrogenase (DPD) catabolizes 5-FUra to an inactive molecule. The susceptibility of tumors to fluoropyrimidines is reported to correlate with tumor levels of these enzymes. To obtain some insight into the tumor types susceptible to fluoropyrimidine therapy, we measured expression levels of these two enzymes in various types of human cancer tissues (241 tissue samples) by the ELISA methods. DPD exists in all the cancer types studied, such as bladder, breast, cervical, colorectal, esophageal, gastric, hepatic, pancreatic, prostate, and renal cancers. Among them, the cervical, hepatic, pancreatic, esophageal, and breast cancer tissues expressed high levels of DPD (median >70 U/mg protein), while high concentrations of the dThdPase were expressed in esophageal, cervical, breast, and pancreatic cancers and hepatoma (median >150 U/mg protein). The dThdPase/DPD ratio, which was reported to correlate with the susceptibility of human cancer xenografts to capecitabine, was high in esophageal, renal, breast, colorectal, and gastric cancers (median ratio of >1.5). In any of these three parameters, the inter-patient DPD variability for each cancer type was much larger than the DPD variability among cancer types; highest/lowest ratios for dThdPase, DPD, and dThdPase/DPD were 10-321, 7-513, and 2-293, respectively. These results indicate that measurements of the three parameters, DPD, dThdPase and dThdPase/DPD, would be useful criteria for selecting cancer patients suitable for fluoropyrimidine therapy rather than for selecting cancer types.

Animals↗

PubMed2000

Candida albicans: adherence, signaling and virulence.

The focus of this symposium was to present new information on the morphogenesis of Candida albicans, particularly how it relates to signal transduction pathways and other genes involved in the regulation of morphogenesis. In addition, we discuss the role of adherence and colonization of the oral cavity by the organism and discuss the role of mannan as an adhesin that recognizes the human red blood cell. C. albicans utilizes at least two signal pathways to regulate its conversion from a yeast form to filamentous growth (hyphae). One of these two pathways is similar to the Saccharomyces cerevisiae pseudohyphal/mating pathway, which utilizes the regulatory protein, Cphlp. The other pathway is not totally defined but requires a second regulatory protein, referred to as Efg1p. Other signal pathways may exist, which include a two-component histidine kinase and response regulator proteins. The latter pathway(s) may include proteins such as Chk1p, Ssk1p, Shi1p and Cos1p/Nik1p. Mutations in strains, which specifically target these proteins, result in morphogenesis defects and avirulence or attenuation of strains. A growth regulatory gene has also been recently defined whose expression is associated with growth cessation and which appears to be a necessary prerequisite in conversion of the organism to a filamentous growth form. Starvation of yeast cells induces exponentially grown cells (and usually non-germinative) to germinate. This phenomenon is also observed in cells that are transiently treated with metabolic inhibitors. During each of these treatments (starvation, metabolic inhibition), expression of a growth regulatory gene (CGRI) increases. Adherence of C. albicans to host cells and tissues is complex; several proteins, which appear to have host recognition functions, have been defined. In the oral cavity, C. albicans selectively adheres to salivary proteins, which are absorbed to many oral surfaces. This mechanism enables the cells to colonize surfaces of the oral cavity. An understanding of these interactions may lead to strategies to prevent oral disease. Mannan from C. albicans may provide a host recognition function for C. albicans. Recent experiments indicate that mannan binds to human red blood cells and causes hemolysis. Binding of mannan to the band 3 protein of human red blood cells has been established. This activity may be associated with the ability of the organism to utilize hemoglobin (and iron).

Candida albicans↗

PubMed1999

Physical training augments plasma catecholamines and natural killer cell activity.

To elucidate the effects of a 10-week exercise training period on physical fitness and plasma catecholamine concentration at rest, and on natural killer cell activity in young healthy untrained females, subjects (20-22 years old. N = 16) carried out physical training by a protocol which consisted of treadmill jogging at a work-intensity of 50% of their VO2max, two hours a day three times a week for ten weeks. VO2max was increased significantly from 33.1 +/- 3.4 ml/kg/min to 38.3 +/- 6.6 ml/kg/min by the ten weeks of physical training (P < 0.005), and natural killer cell activity was also increased significantly from 31.9 +/- 14.3% to 46.4 +/- 18.4% (P < 0.05). The concentrations of epinephrine and norepinephrine before and after physical training were 18.3 +/- 8.7 pg/ml and 20.4 +/- 8.9 pg/ml, and 134.1 +/- 52.2 pg/ml and 248.1 +/- 106.8 pg/ml (P < 0.005), respectively. Changes in norepinephrine and epinephrine correlated with the change in VO2max (r = 0.780, P < 0.005; r = 0.556, P < 0.05). While the change in natural killer cell activity correlated with the change in epinephrine (r = 0.623, P < 0.01), the correlation of the change in natural killer cell activity with change in norepinephrine did not reach statistical significance (r = 0.497, P = 0.0503). From these results we concluded that physical training augments plasma catecholamine levels and natural killer cell activity at rest in young healthy females.

Adult↗

PubMed1999

[Procainamide-induced skin eruption associated with disseminated intravascular coagulation in a patient with sustained ventricular tachycardia].

A 70-year-old man, who had sustained ventricular tachycardias following a previous anterior myocardial infarction, suffered from skin eruptions and abnormal blood tests after 10 days following the oral administration of 1500 mg/day of procainamide. These abnormalities disappeared early after the discontinuation of oral procainamide. However, similar skin eruptions exhibited again when the procainamide was resumed. These results suggest that oral procainamide therapy induces skin eruptions and serious abnormal blood tests in the patient. No reports have suggested such a serious early complication by procainamide therapy. Careful follow-up is needed after the administration of oral procainamide therapy.

Aged↗

PubMed1999

Structural requirements of a human deoxyribonuclease II for the development of the active enzyme form, revealed by site-directed mutagenesis.

Using site-directed mutagenesis, we eliminated three potential N-glycosylation sites (N86, N212, and N266) of human deoxyribonuclease II (DNase II), conserved in mammalian enzymes, and a proteolytic processing site (Q46-R47), forming a propeptide subunit of the enzyme. We expressed a series of these mutant DNase II constructs in COS-7 and Hep G2 cells. Liberation of each glycosylation site at N86 and N266 and the cleavage site interfered dramatically with expression of the intracellular and secreted DNase II activities, irrespective of cell line transfected. A chimeric mutant in which the signal peptide of the DNase II was replaced with that of human DNase I had no intracellular or secreted enzyme activity. Therefore, a simultaneous attachment of a carbohydrate moiety to N86 and N266, cleavage of the propeptide from the single DNase II precursor, and the inherent signal peptide might be required for subcellular sorting and proteolytic maturation of the enzyme.

Amino Acid Substitution↗

PubMed1999

Antioxidant function of the mitochondrial protein SP-22 in the cardiovascular system.

The mitochondrial protein SP-22 has recently been reported to be a member of the thioredoxin-dependent peroxide reductase family, suggesting that it may be one of the antioxidant systems in mitochondria, which are the major site of reactive oxygen intermediate generation. The aim of this study was to examine whether SP-22 is involved in mitochondrial antioxidant mechanisms and whether its expression is induced by oxidative stresses, particularly those in mitochondria. The expression of SP-22 protein was enhanced by about 1.5-4.6-fold when bovine aortic endothelial cells (BAEC) were exposed to various oxidative stresses, including mitochondrial respiratory inhibitors which increased the superoxide generation in BAEC mitochondria. The expression of SP-22 mRNA increased 2.0-3.5-fold with a peak at 3-6 h after exposure to Fe2+/dithiothreitol or a respiratory inhibitor, antimycin A. BAEC with an increased level of SP-22 protein caused by pretreatment with mild oxidative stress became tolerant to subsequent intense oxidative stress. On the other hand, BAEC that had been depleted of SP-22 with an antisense oligodeoxynucleotide against SP-22 mRNA became more labile to oxidative stress than control BAEC. The induction of SP-22 protein by oxidative stress in vivo was demonstrated in an experimental model of myocardial infarction in rat heart. These findings indicate that SP-22 functions as an antioxidant in mitochondria of the cardiovascular system.

Amino Acid Sequence↗

PubMed1999

Characteristics of genomic breakpoints in TLS-CHOP translocations in liposarcomas suggest the involvement of Translin and topoisomerase II in the process of translocation.

Fusion of TLS/FUS and CHOP gene by reciprocal translocation t(12;16)(q32;q16) is a common genetic event found in myxoid and round-cell liposarcomas. Characterization of this genetic event was performed by three methods, Southern blot, RT-PCR, and genomic long-distance PCR in nine myxoid and three round-cell liposarcomas. All but one tumors showed genetic alternations indicating the fusion of TLS/FUS and CHOP gene. Two novel types of fusion transcripts were found, of which one lacked exon 2 sequence of CHOP gene, and the other lacked 3' half of exon 5 of TLS gene. The latter case was caused by a cryptic splicing site which was created by the genomic fusion. Detailed analyses genomic fusion points revealed several sequence characteristics surrounding the fusion points. Homology analyses of breakpoint sequences with known sequence motifs possibly involve in the process of translocation uncovered Translin binding sequences at both of TLS/ FUS and CHOP breakpoints in two cases. Translocations were always associated with other genetic alterations, such as deletions, duplications, or insertions. Short direct repeats were almost always found at both ends of deleted or duplicated fragments some of which had apparently been created by joining of sequences that flank the rearrangement. Finally, consensus topoisomerase II cleavage sites were found at breakpoints in all cases analysed, suggesting a role of this enzyme in creating staggered ends at the breakpoint. These data suggested that sequence characteristics may play an important role to recruit several factors such as Translin and topoisomerase II in the process of chromosomal translation in liposarcomas.

Artificial Gene Fusion↗

PubMed1999

Diffuse bilateral subacute neuroretinitis: first patient with documented nematodes in both eyes.

OBJECTIVE: To describe the first patient with documented evidence of diffuse unilateral subacute neuroretinitis (DUSN) in both eyes. METHODS: A 10-year-old healthy Brazilian girl was first seen with signs of late-stage DUSN in both eyes. A careful search for a nematode was performed in each eye. RESULTS: A motile 550- to 660-microm nematode was found in the inferotemporal retina of the left eye. A similar-sized motile nematode was found in the superotemporal retina of the right eye. Both nematodes were treated with argon green laser applications with bilateral improvement of visual function. CONCLUSION: Although most patients with DUSN do not develop the disease in the fellow eye, this case demonstrates that DUSN can occasionally affect both eyes.

Acute Disease↗

PubMed1999

Chemotaxis and activation of particle-challenged human monocytes in response to monocyte migration inhibitory factor and C-C chemokines.

Cytokines that regulate monocyte migration were found in membrane tissue surrounding loosened prosthetic implants. Monocyte migration inhibition factor (MIF) is able to inhibit macrophage migration. Monocyte chemoattractant protein (MCP) and macrophage inflammatory protein (MIP) are potent macrophage chemoattractants. These cytokines may be expressed as part of the foreign body response to prosthetic particulate debris. Chemotaxis analysis and macrophage activation experiments were performed to determine the effects of MIF, MCP-1, and MIP-1alpha on macrophage migration and activation in vitro. We demonstrated that MIF had its maximal migration inhibitory effect for unchallenged and particle challenged macrophages at 1 ng/mL. MCP-1 and MIP-1alpha stimulated macrophage chemotaxis maximally at 1 to 10 ng/mL. Dose-response studies with MIF, MCP-1, and MIP-1alpha demonstrated that these cytokines did not modulate activation of unchallenged or particle challenged macrophages as evaluated by IL-6 and TNF-alpha release. However, these cytokines do not appear to affect macrophage release of proinflammatory mediators in vitro.

Biocompatible Materials↗