Biomedical subjects
Publications and source records attributed to K Nishigaki.
A rapid 5'-labeling method of single-stranded DNA/RNA was developed, which is based on the utilization of an adenylated intermediate in the reaction of T4 RNA ligase. This method is commonly useful for fluorescence-, isotope- or biotin-labeling of the 5'-ends of both oligo- and polynucleotides.
Neoplastic disease, especially malignant lymphomas, are often observed in cats infected with feline immunodeficiency virus (FIV). In order to clarify the characteristics of lymphoma cells and to investigate the pathogenesis in FIV-infected cats, we examined the lymphoma tissues developed in five cats naturally infected with FIV by Southern blot analyses using feline immunoglobulin (Ig), T-cell receptors (TCR) and FIV probes. All of the five cases were serologically positive for anti-FIV antibody and negative for feline leukemia virus antigen. Of these five lymphoma samples, two displayed rearrangement of the Ig heavy chain gene and deletion of the Ig light (kappa) chain gene, indicating that the tumor cells were committed to B-cell development. One tumor sample was identified as a T-cell lymphoma because of the presence of a rearranged TCR beta-chain gene. The other two cases were considered to be non-T non-B cell lymphoma because they did not show any rearrangement of the Ig and TCR genes. Therefore, no consistent tumor type was found in lymphoma cases infected with FIV. Clonal integration of FIV provirus was not detected in any of the five lymphoma samples obtained from FIV-infected cats using Southern blot analysis, although FIV proviral genome was detected in the genomic DNA of all the lymphoma samples by using a polymerase chain reaction (PCR). These results indicated that FIV might not play a direct role in tumorigenesis of lymphoma in cats.
OBJECTIVES: This study sought to examine plasma levels of soluble Fas/APO-1 receptor (sFas), an inhibitor of apoptosis, and soluble Fas ligand (sFas-L), an inducer of apoptosis, and their relation to each other and to other clinical variables, such as New York Heart Association functional class, tumor necrosis factor (TNF) and interleukin-6 (IL-6) in congestive heart failure (CHF). BACKGROUND: It has been recently reported that apoptotic cell death occurs in myocytes of dogs with CHF. Hypoxia is frequently seen in advanced CHF and can stimulate Fas/APO-1 receptors (Fas) to induce apoptosis in cultured myocytes. Fas and Fas ligand (Fas-L) are cell-surface proteins and representative apoptosis-signaling molecules. Fas on the cell membrane induces apoptosis when it binds Fas-L or sFas-L. However, plasma sFas, a molecule lacking the transmembrane domain of Fas, blocks apoptosis by inhibiting binding between Fas and Fas-L or sFas-L on the cell membrane. At present, it is unknown whether plasma sFas-L and plasma sFas increase in the presence of cardiac disease. METHODS: The study included 70 patients (mean [+/-SEM] age 65 +/- 2 years, range 21 to 93) with chronic CHF (coronary artery disease in 28, dilated cardiomyopathy in 27, valvular heart disease in 15) and 62 age- and gender-matched normal control subjects. Plasma levels of sFas, sFas-L, TNF-alpha and IL-6 were measured by enzyme-linked immunosorbent assays using monoclonal anti-human antibodies. RESULTS: There was no significant difference in sFas-L levels between normal subjects and patients in functional classes I to IV; however, sFas increased with severity of functional classification, independent of the underlying disease. sFas levels were significantly higher even in patients in functional class II than in normal subjects and those in functional class I, and were highest in patients in functional class IV (normal subjects; 2.2 +/- 0.1 ng/ml; functional class I: 2.2 +/- 0.2 ng/ml; functional class II: 3.1 +/- 0.2 ng/ml; functional class III: 3.9 +/- 0.3 ng/ml; functional class IV: 5.1 +/- 0.6 ng/ml). Plasma sFas levels were significantly higher in patients with elevated pulmonary artery wedge pressure and a decresed cardiac index than in those with values in the normal range. In patients in functional class IV, there was no significant difference in plasma sFas levels between the survivors and non-survivors during 6-month follow-up. However, plasma levels of sFas tended to decrease in nine patients with clinical improvement (baseline sFas: 5.2 +/- 0.8 ng/ml; 6-month sFas: 4.3 +/- 0.5 ng/ml, p = 0.07) but were similar in patients with no change in functional class. TNF-alpha and IL-6 were increased significantly only in patients in functional class IV, as previously reported, but were not related to sFas. CONCLUSIONS: We found elevated levels of plasma sFas and no increase in plasma sFas-L in human CHF. The increase in sFas may play an important role in the pathophysiologic mechanisms of CHF.
ATP-dependent enzymes were investigated as to the stringency of their ATP requirement. For all the enzymes examined except firefly luciferase (including hexokinase, polynucleotide kinase, T4 DNA ligase, and T4 RNA ligase) ATP could be replaced with dATP, contradicting previous data. Considering the replaceable nucleotides, not only kinases (low stringency as to ATP-requirement) but also other enzymes (moderate stringency) were typed as phosphate-directed ATP recognition. Through this study, an exact view of ATP-requiring enzymes which have a profound influence on the concentration in a cell of ATP, a metabolic and regulative key substance, was obtained, and a technically useful, fluorescent ATP-substitute (2AP-TP) was introduced.
Long terminal repeats of feline leukemia viruses cloned from feline acute myeloid leukemias frequently contained direct repeats of 40 to 74 bp in the upstream region of the enhancer (URE). The repetitive URE conferred an enhancer function upon gene expression in myeloid cells, suggesting its association with tumorigenic potential in myeloid cells.
A 17-year-old woman was found to have ectopic atrial tachycardia by her physician. Echocardiography and cardiac catheterization revealed findings resembling dilated cardiomyopathy at the time of initial presentation. The tachycardia was controlled with atenolol only at a dose of 50 mg/day. However, at the age of 22, the presence of ectopic atrial tachycardia was once again confirmed. We successfully performed catheter ablation for persistent ectopic atrial tachycardia. Serial echocardiographic findings showed the left ventricular dimension and function appeared to return to normal 1 year postablation. However, despite pharmacologic control and catheter ablation therapy, histopathology revealed myocardial fibrosis presumably representing permanent damage of the heart secondary to tachycardia 1 year postablation.
This study examined the effects of Albunex (sonicated 5% human serum albumin) infusion on left ventricular inflow velocity by Doppler echocardiography. Left ventricular pressure and left ventricular inflow velocity were recorded simultaneously under eight different conditions in dogs: 1) baseline 1 (control), 2) Albunex 0.2 ml/kg, 3) baseline 2, 4) Albunex 0.5 ml/kg, infusion of dextran 100 ml, 5) baseline 3, 6) Albunex 0.2 ml/kg, 7) baseline 4, and 8) Albunex 0.5 ml/kg. In the normal state (no dextran), Albunex (0.2 ml/kg) caused no hemodynamic changes or inflow velocity changes. In contrast, infusion of Albunex (0.5 ml/kg) caused time velocity integrals of early filling to increase from the baseline (5.51 +/- 1.13 vs 7.19 +/- 1.14 cm, p < 0.05). After dextran infusion (100 ml), Albunex (0.2 ml/kg) caused peak early filling velocity to increase (62.4 +/- 6.9 vs 67.3 +/- 9.4 cm/sec, p < 0.05), and infusion of Albunex (0.5 ml/kg) also caused peak early filling velocity to increase from baseline (64.6 +/- 8.5 vs 73.7 +/- 14.5 cm/sec, p < 0.05). Infusion of Albunex (0.5 ml/kg) after dextran infusion caused increases in left ventricular pressure at the mitral valve opening (12.7 +/- 3.1 vs 15.2 +/- 3.3 mmHg, p < 0.05) and in left atrial driving force (13.5 +/- 3.6 vs 16.7 +/- 5.9 mmHg, p < 0.05). Clinicians should be cautious about using Albunex at doses of greater than 0.2 ml/kg when evaluating the pressure gradient of the left ventricle in patients with elevated left ventricular diastolic pressure. In patients with normal hemodynamics, Albunex infusion at doses of less than 0.2 ml/kg apparently did not affect the velocity measurement.
Repetitive structure of enhancer elements in the long terminal repeat (LTR) has been identified in feline leukemia viruses (FeLVs) integrated in lymphoid tumor cells in cats. In this study, promoter activities of the FeLV LTRs were measured in lymphoid and non-lymphoid cell lines in transient expression assays using plasmids containing the viral LTRs linked to the chloramphenicol acetyltransferase (CAT) gene. Promoter activity of the LTR with 3 enhancer repeats (pFTLTR) was significantly higher than that of the LTR with 1 enhancer (Glasgow-1 LTR) in feline (FT-1) and human (Jurkat) T-lymphoblastoid cell lines. Promoter activity of the pFTLTR was also significantly higher than that of its mutant (pFTLTR1E) in which 2 of the 3 enhancers were deleted in FT-1 and Jurkat cells. Both of these differences were not observed in a feline fibroblastic cell line (CrFK). Moreover, mutations affecting the consensus motifs for LVb, SV40, NF-1, GRE and FLV-1 resulted in decreased basal activity of the FeLV LTR (pFTLTR1E) in FT-1, Jurkat and CRFK cells. The decrease of the promoter activity was especially remarkable in FT-1 cells. The present study revealed the strong promoter activity of the FeLV LTR with 3 enhancer repeats and its modular enhancer elements positively regulating the transcription in a relatively tissue-specific manner.
Left atrial (LA) operative mean stiffness was measured using simultaneous recordings of the left ventricular (LV) pressure and M-mode echocardiography of the LA and mitral valve. The LA operative passive mean stiffness value was obtained during LV systole using LV pressure at the mitral valve opening and the pre-atrial contraction where the LV and LA pressure curves cross each other. Before the LA stiffness measurement, the LA volume calculated by biplane left atriography was compared with the dimension of the LA M-mode echocardiogram at three points (maximum volume, pre-atrial contraction and minimum volume) in another 23 patients (5 normal subjects, 4 patients with angina pectoris, 14 patients with myocardial infarction), and the regression equation was obtained by power fitting (y = ax3+ b). Using this equation, the LA volumes were calculated and used for the measurement of LA operative mean stiffness. Eleven normal subjects (C group), 14 patients with myocardial infarction (ejection fraction: EF > or = 55%; NF group), and 12 patients with myocardial infarction (EF < 55%; F group) were studied. The measured operative mean stiffness values based on the LA dimension and LV pressure [K(D)] were 0.69 +/- 0.40, 1.0 +/- 0.37, and 2.0 +/- 0.61 mmHg/mm, respectively (p < 0.01 in C vs F). The mean stiffness values calculated with LA volume [K(V)] were 0.48 +/- 0.23, 0.42 +/- 0.19, and 0.66 +/- 0.25 mmHg/ml, respectively (p < 0.05 in NF vs F). In F group, both the K(D) and K(V) values were high. The K(D) value can thus be used clinically as an easily obtained index of the LA operative mean stiffness. The high LA operative mean stiffness in F group appeared to be related to the increased LA pressure at the mitral valve opening. This method of measurement of the LA operative mean stiffness can be easily applied and used as a routine measurement providing additional information regarding left ventricular function.
OBJECTIVES: We examined whether plasma brain natriuretic peptide levels are abnormally elevated in hypertrophic obstructive cardiomyopathy compared with other cardiac diseases. BACKGROUND: We previously reported that plasma brain and atrial natriuretic peptide levels were elevated in hypertrophic cardiomyopathy. METHODS: We compared plasma concentrations of brain and atrial natriuretic peptide and hemodynamic and echocardiographic data in 50 patients with hypertrophic obstructive cardiomyopathy (n = 15, mean [+/-SD] intraventricular pressure gradient 37 +/- 16 mm Hg), hypertrophic nonobstructive cardiomyopathy (n = 15), aortic stenosis (n = 10, mean pressure gradient 41 +/- 18 mm Hg) and hypertensive heart disease (n = 10, mean systolic/diastolic blood pressure 203 +/- 16/108 +/- 11 mm Hg, respectively) and 10 normal subjects. RESULTS: Plasma brain natriuretic peptide levels were higher in the hypertrophic obstructive cardiomyopathy group (397.1 +/- 167.8 pg/ml*) than in the hypertrophic nonobstructive cardiomyopathy (60.0 +/- 48.1 pg/ml*), hypertensive heart disease (53.9 +/- 31.4 pg/ml*), aortic stenosis (75.4 +/- 54.3 pg/ml*) and normal groups (9.8 +/- 6.4 pg/ml [*p < 0.05 vs. normal group, p < 0.05 vs. hypertrophic obstructive cardiomyopathy group]). Although plasma atrial natriuretic peptide levels were higher in the hypertrophic obstructive cardiomyopathy group than the other patient groups, the brain/atrial natriuretic peptide ratio in the hypertrophic obstructive cardiomyopathy group was higher (4.5 +/- 2.3) than those in the other three patient groups (1.1 to 1.4) and the normal group (0.7 +/- 0.5). Left ventricular end-diastolic pressure and left ventricular end-diastolic volume index were similar among the four patient groups. The interventricular septal thickness and the ratio of interventricular septal thickness to left ventricular posterior wall thickness were similar between the hypertrophic obstructive and nonobstructive cardiomyopathy groups. CONCLUSIONS: Abnormal elevations of plasma brain natriuretic peptide levels are difficult to explain on the basis of hemodynamic and echocardiographic data and are a special feature of hypertrophic obstructive cardiomyopathy.
BACKGROUND AND HYPOTHESIS: Flow velocity of the left atrial appendage (LAA) is thought to be important in thrombus formation in association with blood stasis and the development of spontaneous echo contrast. The effects of heart rate on peak flow velocity of LAA have not been studied in patients with nonvalvular atrial fibrillation. METHODS: Using transesophageal Doppler echocardiography, peak flow velocity of the LAA was measured at the junction between the left atrium and the LAA during left ventricular (LV) systole and diastole in 21 patients with nonvalvular atrial fibrillation. In six cases, the average peak flow velocity of the LAA for 10 consecutive beats with moderately long R-R intervals (LI beats) was compared with those of 3-5 consecutive beats with extremely short R-R intervals (SI beats). RESULTS: Average peak flow velocity of the LAA during LV diastole was significantly higher than that during LV systole (26.5 +/- 15.7 vs. 19.3 +/- 10.4 cm/s, p < 0.01). In SI beats, average peak flow velocity of the LAA was significantly lower than that in LI beats (17.1 +/- 12.1 vs. 21.2 +/- 12.9 cm/s, p < 0.01). CONCLUSION: An increased heart rate reduced the peak flow velocity of the LAA in patients with nonvalvular atrial fibrillation, which would promote blood stasis in the LAA.
Atrial reservoir function has not been studied after successful cardioversion of chronic atrial fibrillation. Using transthoracic and transesophageal Doppler echocardiography, we measured flow velocity-time integrals of the systolic forward (Sa), diastolic forward (Da), and diastolic reversed (rAa) waves of flow velocity waveforms in the pulmonary vein and the superior vena cava, and those of the early diastolic (Ea) and late diastolic (Aa) waves of the transmitral and transtricuspid flow velocity waveforms. The left and right atrial storage fractions (LASF, RASF), indexes of atrial reservoir function, were determined as the ratios of the atrial storage volume to the ventricular stroke volume; (Sa - rAa)/(Sa - rAa + Da). The left and right atrial active contraction fractions (LAACF, RAACF), indexes of atrial active transport function, were also determined as the ratios of the atrial active contraction volume to the left ventricular stroke volume; Aa/(Ea + Aa). These indices were evaluated periodically in 12 patients with non-valvular chronic atrial fibrillation before and 1-4 days after direct current cardioversion of atrial fibrillation; in 8 of the patients, the indices were also evaluated 1-3 months after the cardioversion. An additional 10 patients in sinus rhythm served as controls. Both the LASF and RASF were low during atrial fibrillation; the values increased significantly 14 days after successful cardioversion (P < 0.01 P < 0.01), and continued to increase at 1-3 months. The LASF and RASF values 1-3 months after cardioversion were comparable to those in control subjects. Both the LAACF and RAACF also increased significantly from 1-4 days to 1-3 months after cardioversion (P < 0.05, P < 0.01), becoming comparable to those in control subjects. During the 3 months after successful cardioversion of non-valvular chronic atrial fibrillation, left and right atrial reservoir function and left and right atrial active transport function increased progressively, becoming comparable to values in the control subjects.
To extend the indications for corrective operation in patients with pulmonary atresia, ventricular septal defect, and major aortopulmonary collateral arteries, surgical procedures were done to unify the blood sources for pulmonary perfusion. Since December 1985, 50 patients have undergone unifocalization at ages from 2 months to 26 years with a mean of 6 +/- 7 years. In total, 84 staged unifocalization procedures and 5 other palliative procedures were done in 49 patients. These included several operative procedures: simple ligation of major aortopulmonary collateral arteries in 8; pulmonary angioplasty in 29 including reconstruction of the pulmonary arterial tree by direct anastomosis or interposition between the central pulmonary arteries and the intrapulmonary arteries; construction of artificial central pulmonary arteries with use of a xenograft pericardial tube graft in 36 with no native central pulmonary arteries detected; and construction of supplemental central pulmonary arteries also with use of a pericardial tube graft in 10. The pericardial tube graft, if used, was anastomosed to the intrapulmonary arteries on one end and connected to a prosthetic tube on the other end so as to perfuse the reconstructed pulmonary arteries. The anastomosis was made inside the lung through the divided interlobar fissure. Five patients died after operation among those undergoing these 89 preparative operative procedures. Deaths were related either to bleeding caused by anticoagulation therapy administered to prevent thrombosis within the xenograft pericardial tube graft used or to progressive congestive heart failure as a result of an excessive amount of pulmonary blood flow. Twenty-six patients have undergone intracardiac repair after previous unifocalization. In 16 patients the artificial central pulmonary arteries surgically constructed were connected to each other and then an external conduit was placed. In another patient, intracardiac repair and unifocalization could be concomitantly achieved via a median sternotomy. The right ventricle to left ventricle systolic pressure ratio immediately after intracardiac repair in 27 patients ranged from 0.24 to 0.91 with a mean of 0.54 +/- 0.17. One patient (4%) died shortly after intracardiac repair because of thrombosis within the pulmonary arteries. Postoperative catheterization showed that pulmonary vascular resistance was correlated significantly with the number of pulmonary vascular segments functioning rather than with the condition of the central pulmonary arteries. We conclude that surgical unifocalization is a feasible procedure before subsequent intracardiac repair, even in patients with critically hypoplastic or absent central pulmonary arteries.
Unifocalization, a surgical technique to unifocalize the pulmonary blood supply in patients with pulmonary atresia, ventricular septal defect and major aortopulmonary collateral arteries, is a useful preparative operation to extend the indication for corrective surgery. The preoperative and postoperative pulmonary angiograms of 51 patients (aged 3 months-26 years at first unifocalization, 29 males, 22 females), who underwent a total of 96 unifocalizations from December 1985 to July 1994, were studied to assess the effectiveness of each procedure of unifocalization. The procedures of unifocalization were ligation (9), angioplasty (6), direct anastomosis (25), bridging (6), additional central pulmonary artery creation (9), central pulmonary artery creation (36) and thrombectomy (5). Altogether 125 anastomoses were made, and the patency of 101 (80.2%) anastomoses was confirmed, 24 (19.2%) were shown to be occluded. The patency rate of the anastomoses between xenograft rolls and intrapulmonary arteries was 79.5% (70/88), while that between native central pulmonary arteries and intrapulmonary arteries was 83.8% (31/37; N. S.). The patency rate of the anastomoses involving intrahilar arteries was 88.0% (44/50), while that for the group involving extrahilar arteries only was 71.9% (46/64; P < 0.05). It is concluded that 1) unifocalization with the reconstruction of central pulmonary arteries using a pericardial roll is a useful method for patients with absent or hypoplastic central pulmonary arteries, 2) it is preferable to divide the fissures of lung in anastomosing pulmonary arteries of arborization abnormalities.
The cause of separation in gel electrophoresis between highly homologous ss (single-stranded) DNAs as observed in SSCP (single-strand conformation polymorphism) was pursued. Advancing our previous explanation [J. Biochem. 99, 663-671 (1986)], the mobility difference of ssDNAs was correlated to differences in their dynamic conformation (not to differences in their most stable structure), focusing on point-substituted sites. The contribution of semistable conformation dynamics was considered to be critical. Putative factors which may influence mobility (i.e., length of ssDNAs, location of substituted sites, and types of substitutions) were experimentally examined and critically discussed. Understanding of these phenomena should yield improvements in various techniques, such as SSCP, and in evaluation of the solution structures of DNA.
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Between November 1987 and January 1990, 33 patients (tricuspid atresia, 9 patients; mitral atresia, 3; single ventricle, 15; others, 6) underwent Fontan operations. The rate of blood flow returning to the heart during aortic cross-clamping was measured as an indication of the extent of development of aortopulmonary collateral arteries. Percent cardiac return (calculated by dividing the blood flow rate returning to the heart by the cardiopulmonary bypass blood flow rate and expressing the value as a percentage), were 1% to 9%, 7 patients; 10% to 19%, 11; 20% to 29%, 9; 30% to 39%, 4; 40% to 49%, 1; and 50% to 59%, 1 patient. Percent cardiac return showed a significant correlation with postoperative mean systemic venous pressure (r = 0.6, p < 0.01). In those patients in whom percent cardiac return was more than 33%, the mean systemic venous pressure after operation was high (more than 17 mm Hg), and none of these patients survived. To predict percent cardiac return preoperatively, the conventional indices of systemic ventricular volume, pulmonary artery area index, arterial blood oxygen saturation, pulmonary blood flow index, and pulmonary vascular resistance were analyzed. None of these showed significant correlation with percent cardiac return. However, all the patients who had a high percent cardiac return (more than 30%) also had both high arterial blood oxygen saturation (more than 75% in room air) and small pulmonary artery area index (less than 55%). In addition, the age at operation showed good correlation (r = 0.6, p < 0.01) to percent cardiac return.(ABSTRACT TRUNCATED AT 250 WORDS)