[Hepatitis C virus genotyping by restriction fragment length polymorphism of polymerase chain reaction products generated with a HCV detection kit].

Highly conserved sequence in the 5' untranslated region(UTR) of hepatitis C virus(HCV) genome have been targeted by most nucleic acid amplification-based detection assays, such as Amplicor HCV test, a commercially available assay kit. In this study, we classified HCV genotypes by direct sequencing determination for 5' UTR of nested-PCR after Amplicor HCV test. Then, based on the results of sequence, RFLP analysis after digestion of the nested PCR fragments with Hae III or Sau 3AI to classify HCV genotype was evaluated. RFLP analysis distinguished the type 1, 2a and 2b. Only one of 29 samples was not classified by RFLP analysis due to the point mutation of Hae III recognition site. HCV genotypes commonly found in JAPAN were classified into three types, 1b, 2a, and 2b. Also, RFLP analysis requires fewer resources than serotype grouping test. Hence, the present method provides an adaptable and rapid HCV genotyping in clinical laboratory in JAPAN.

[Hepatitis B virus gene mutations in the sera of three patients with coexisting hepatitis B surface antigen and anti-surface antibody].

We analyzed gene mutations in the Hepatitis B virus of three virus carriers with coexisting Hepatitis B surface(HBs) antigen and anti-HBs antibody. Viral DNAs were extracted from sera and the pre-S, S and X(including core promoter and pre-core region) regions were amplified by PCR, and sequenced. Case 1 and Case 2 were positive for HBe antigen, while Case 3 was negative. All three cases were positive for HBe antibody and HBV DNA. In the S gene region, various point mutations were detected in all three cases. Mutations were clustered in the first hydrophilic loop region(codon 47-46) essential for the secretion of surface antigen. A few mutations were detected in 'a' loop(codon 124-147) of the S gene. None of the cases had an amino acid substitution of codon 145 of the S gene that is reported to be responsible for weak recognition by the HBs antibody. These data suggest the existence of hyper-variable sequence in S region, or otherwise result of low-fidelity of Taq DNA polymerase-reaction. Case 1 possessed a point mutation, T to C at nucleotide position 1753, in the region overlapping the coding region of the X gene and the CCAAT/enhancer binding protein(C/EBP) binding region within the core promoter region. Case 2 possessed both a large deletion(129 bp) in the pre-S1 and in-frame deletions of 15 and 27 bp in the pre-S2 region. Case 3 had an in-frame deletion of 30 bp in the pre-S2 region, and a point mutation in precore region. The point mutation, G to A at a nucleotide position 1986, converts Trp(TGG) to a stop codon TAG, and may contribute the fulminant hepatitis. These results suggest that the mutations in the pre-S, the core promoter, or the X gene may imply coexistence of the HBs antigen and antibody after seroconversion, while the point mutations in the S region are not likely to be responsible for the HBV escape mutant.

[Van der Hoeve's syndrome with Ebstein's anomaly, and prolapse of the mitral and aortic valves: a case report].

A hitherto unrecognized case of van der Hoeve's syndrome complicated by Ebstein's anomaly, and prolapse of the mitral and aortic valves was reported. A 46-year-old woman presented with blue sclera, osteogenesis imperfecta and a hearing loss, which are typical symptoms of this syndrome. The electrocardiogram showed a type B WPW syndrome. The phonocardiogram showed a loud and widely split first heart sound, an accentuated protodiastolic extrasound, a decrescendo regurgitant systolic murmur, and a presystolic murmur. The x descent of the jugular phlebogram was obliterated by a markedly increased c wave. Based on M-mode and two-dimensional echocardiograms, 1) the interventricular septal motion was paradoxical and the closure of the tricuspid valve was delayed, 2) the septal tricuspid leaflet was displaced toward the apex from its normal annular insertion on the apical four-chamber view, 3) the three leaflets of the tricuspid valve were abnormally elongated, 4) the anterior mitral leaflet and the non-coronary cusp of the aortic valve were elongated and prolapsed. Doppler echocardiography detected severe tricuspid and mild mitral regurgitations. We suggest that the development of Ebstein's anomaly is possibly related to that of osteogenesis imperfecta genetically and that not only Ebstein's anomaly but a connective tissue disorder contributes to the elongated and prolapsed leaflets.

[Apical mid-diastolic rumble in hypertrophic cardiomyopathy: a pulsed Doppler echocardiographic study].

To investigate the mechanism of an apical mid-diastolic rumble in hypertrophic cardiomyopathy (HCM), we recorded left ventricular (LV) inflow velocity patterns using pulsed Doppler echocardiography and apexcardiography for 10 HCM patients with rumble and 20 HCM patients without rumble. Controls consist of 17 normal subjects, three patients with complete atrioventricular block and two patients with artificial right ventricular pacemakers. The LV inflow velocity profiles were analyzed in terms of acceleration time (AT) and deceleration time (DT) of the rapid filling wave, and the ratio of peak velocity of the atrial contraction wave to that of the rapid filling wave (A/D ratio). The results were as follows: The apical mid-diastolic murmur in HCM had a crescendo-decrescendo character mainly of medium frequency, and increased in intensity after the inhalation of amyl nitrite. All patients with rumble had asymmetric septal hypertrophy and the five of these had LV outflow obstruction. In six of the 10 patients with rumble, mild mitral regurgitation was detected. In HCM with rumble, the AT tended to be shorter than that of HCM without rumble, but it was significantly longer than the AT of normal subjects. In HCM with rumble, the DT was significantly shorter than that of HCM without rumble, but it was significantly longer than the DT of normal subjects. There was no significant difference in the A/D ratio between the HCM with rumble and the normal subjects, but the A/H ratio of the apexcardiogram was significantly increased in HCM with rumble as compared with those of HCM without rumble and of the normal subjects. The LV dimension was significantly decreased in HCM with rumble as compared with those of HCM without rumble and the normal subjects. Peak negative VCF was significantly decreased in HCM with rumble as compared with that of HCM without rumble. But there was no significant difference in this parameter between HCM with rumble and the normal subjects. In simultaneous recordings of apical mid-diastolic rumble and LV inflow velocity patterns, the rumble appeared to start after the beginning of the diastolic rapid filling wave and to stop before or at the end of the diastolic rapid filling wave. In patients with complete atrioventricular block and with artificial right ventricular pacemakers, the apical mid-diastolic rumble appeared when the P wave was during the rapid filling phase of the left ventricle.(ABSTRACT TRUNCATED AT 400 WORDS)

[Stress pulsed Doppler echocardiography in the evaluation of diastolic left ventricular function].

The effect of sublingual nifedipine on left ventricular (LV) function was assessed by analyzing LV inflow velocity patterns using pulsed Doppler echocardiography (PDE) and radioisotope (RI) angiography in 23 patients with hypertrophic cardiomyopathy (HCM), 13 with hypertensive heart disease (HHD) and 24 with ischemic heart disease (IHD). The results were as follows: LV inflow velocity pattern and M-mode echocardiography after the administration of nifedipine (Study-I). The deceleration time (DT) of the diastolic rapid filling wave was shortened after nifedipine in HCM and HHD, but not in IHD. The acceleration time (AT) of the diastolic rapid filling wave and the A/D ratio (amplitude of an atrial contraction wave/amplitude of a diastolic rapid filling wave) were not significantly changed in all groups. Peak negative Vcf was significantly improved after nifedipine in HCM, but unchanged in the other groups. Fractional shortening and peak Vcf did not change in all groups. LV inflow velocity pattern and RI angiography after nifedipine (Study-II). The mean LV rapid filling rate (V1) based on the LV volume curve by multi-gated blood pool scans increased during nifedipine in HCM and HHD, but other diastolic indices, such as the peak filling rate and 1/3 filling fraction, were unchanged. The DT-V1 relationship during nifedipine shifted toward the right and downward in HCM and HHD. In IHD, however, this relationship shifted toward the left and downward in cases with reduced LV function (V1 less than 1.0 EDC/sec), but toward the right and downward in patients whose LV function was maintained (V1 greater than or equal to 1.0 EDC/sec). Nifedipine therapy was associated with an improved LV filling in nearly all cases of HCM and HHD, and in some cases of IHD. PDE and RI angiography are useful for evaluating LV diastolic function during sublingual nifedipine in various heart diseases.

[Left ventricular function in patients with right ventricular overload evaluated by exercise echocardiography: comparison of pulmonary hypertension and atrial septal defects].

To evaluate left ventricular function in patients (pts) with right ventricular overload, exercise echocardiographic studies using a bicycle ergometer were performed for seven pts with pulmonary hypertension (PH), two with cor pulmonale (CP) and 16 with atrial septal defects (ASD), and the results were compared with those of 10 (control I) and 27 (control II) normal persons. These subjects were categorized in two study groups; study I consisting of PH, CP and control group I with an exercise workload of 25 watts for 3 min; and study II consisting of ASD and control group II with an exercise workload of 50 watts for 3 min. The results were as follows: M-mode echocardiography revealed that: During exercise, the stroke volume (SV) was increased with a significant decrease of left ventricular end-systolic dimension (LVDs), but without a change in left ventricular end-diastolic dimension (LVDd) in control groups I and II, with a marked increase of LVDd and a slight decrease of LVDs in the ASD group. By contrast, the SV in the PH group tended to decrease during exercise with a slight decrease of LVDd, but without a significant change in LVDs. Right ventricular dimension (RVD) was significantly increased during exercise in the PH group, but was decreased in the ASD group. In control groups I and II, RVD did not change during exercise. In ASD, and control groups I and II, the peak velocity of circumferential fiber shortening (Vcf) was increased during exercise, and the peak negative Vcf was significantly decreased. However, these parameters exhibited impaired responses during exercise in the PH group. Abnormal interventricular septal (IVS) motion at rest tended to become normal during exercise in 12 of 16 pts with ASD. In all pts with PH, however, IVS motion did not change substantially during exercise. There were similar parameter responses between the PH and CP groups. Two-dimensional echocardiography showed that: The left ventricular short-axis view demonstrated a diastolic left ventricular configuration which changed from oblique to relatively circular orientation during exercise in the ASD group. In four of five pts with PH, the diastolic configuration of the left ventricle was oblique because of a loss of the normal curvature of the IVS at rest and during exercise. The diastolic left ventricular configuration improved during exercise in only one pt with PH, with slightly elevated pulmonary arterial systolic pressure as in the ASD group.(ABSTRACT TRUNCATED AT 400 WORDS)

[Left ventricular diastolic filling patterns in hypertrophic cardiomyopathy and myocardial infarction: studies by pulsed doppler echocardiography and multi-gated blood pool scans].

Relationship between various parameters of the left ventricular inflow velocity patterns by pulsed Doppler echocardiography and the early diastolic mean filling rate (V1) by multi-gated blood pool scans was evaluated. The materials consist of 26 patients with hypertrophic cardiomyopathy (HCM), 24 with old myocardial infarction (MI), seven with hypertensive heart disease (HHD), seven with dilated cardiomyopathy (DCM), seven with ischemic heart disease (IHD), and 16 normal subjects (N). The results were as follows: 1. Acceleration time (AT) and deceleration time (DT) were significantly prolonged in HCM (120 +/- 20 msec, 147 +/- 23 msec), MI (102 +/- 17 msec, 124 +/- 21 msec), HHD (105 +/- 11 msec, 141 +/- 17 msec) and IHD (111 +/- 16 msec, 122 +/- 20 msec) compared with those of normals (89 +/- 20 msec, 106 +/- 18 msec). 2. V1 was significantly decreased in HCM (1.14 +/- 0.28 sec-1), MI (0.68 +/- 0.24 sec-1), HHD (0.73 +/- 0.12 sec-1), DCM (0.67 +/- 0.30 sec-1) and IHD (1.03 +/- 0.29 sec-1) compared with that of normals (1.48 +/- 0.28 sec-1). 3. There were significant negative correlations between V1 and AT or DT in HCM, IHD with slightly impaired diastolic filling and normals. However, there were significant positive correlations between V1 and AT or DT in MI and DCM. These findings suggested that left ventricular diastolic filling is impaired in both HCM and MI, and that DT-V1 relationship is useful for differentiating HCM characterized by the "chamber stiffness" from MI characterized by the "myocardial stiffness".

[Mechanism of an early diastolic posterior motion of the interventricular septum in patients with mitral stenosis: with special reference to left ventricular inflow velocity pattern].

The mechanism of an early diastolic posterior motion of the interventricular septum (IVS) was investigated by means of M-mode, two-dimensional and pulsed Doppler echocardiographies in 53 patients with pure mitral stenosis (MS). Velocity patterns of the blood flow at the inflow tract of the left ventricle (LVIT) were classified into three types as previously reported (cf. Fig.3). The results obtained were as follows: The mitral valve orifice area (MVA) was significantly smaller in patients with type III of the LVIT flow velocity pattern than in patients with type I (p less than 0.001). An amplitude ("a") of the posterior IVS motion in systole (P1) was significantly diminished in type III (4.2 +/- 1.1 mm) than in type I (6.6 +/- 1.5 mm) (p less than 0.001). An amplitude ("b") of the posterior IVS motion in early diastole (P3) showed a significant increase in type III (7.5 +/- 1.2 mm) than in type I (5.2 +/- 1.5 mm) (p less than 0.001). Difference of the amplitude between P1 and P3 ("a-b") was significantly greater in type III (-3.4 +/- 1.1 mm) than in type I (1.4 +/- 0.9 mm) (p less than 0.0001). MVA was correlated statistically with "a" (r = 0.58, p less than 0.001), "b" (r = -0.38, p less than 0.01) and "a-b" (r = 0.80, p less than 0.0001). Deformity of the left ventricular cavity due to the flattened IVS in early diastole was observed in severe MS with an augmented septal P3 dip. A case of severe MS with type III velocity pattern and an augmented P3 dip showed type II velocity pattern and a decreased P3 dip after the attack of acute myocardial infarction. These findings suggested a close relationship between type III of the LVIT flow velocity pattern and the prominent septal P3 dip. The importance of diastolic suction of the left ventricle in producing an early diastolic posterior motion of IVS (P3) was discussed.

Clinical effects of endonasal sinusectomy with reconstruction of the nasal cavity (Takahashi's method).

A follow-up investigation of the results of endonasal sinusectomy with reconstruction of the nasal cavity (Takahashi's method) was carried out in 535 of 1,338 patients who underwent endonasal sinusectomies during the 6-year period from 1967 to 1972. The results of subjective symptoms were as follows: cured 196 (36.6%); improved 234 (47.5%); unchanged 69 (12.9%); aggravated 12 (2.2%). In other words, 450 (84.1%) of the patients were satisfied with the results of the operations. Good results have similarly been obtained in cases of nasal allergy which shows increasing tendency of late, and concomitant reconstruction of the nasal cavity and desensitization therapy have resulted in improved responses.

Clinical effects of endonasal sinusectomy with reconstruction of the nasal cavity (Takahashi's method).

A follow-up investigation of the results of endonasal sinusectomy with reconstruction of the nasal cavity (Takahashi's method) was carried out in 535 of 1338 patients who underwent endonasal sinusectomies during the 6-year period from 1967 to 1972. The subjective results were as follows: cured 196 (36.6%); improved 234 (47,5%); unchanged 69 (12.9%); aggravated 12 (2.2%). In other words, 450 (84.1%) of the patients were satisfied with the results of the operations. Good results have similarly been obtained in allergic nasal disorders spreading in Japan of late, and concomitant reconstruction of the nasal cavity and desensitization therapy have resulted in improved response.