alpha-Fetoprotein as a carrier protein in plasma and its bilirubin-binding ability.

The bilirubin-binding ability of human alpha-fetoproteins, which were purified from fetal cord serum and from ascites fluid of a hepatoma-bearing patient, was examined by the difference spectrum and the Jacobsen peroxidase methods. The difference spectrum observed as a result of the specific binding of bilirubin to alpha-fetoprotein had a maximum at 482 nm, and this pattern was quite similar to that observed for serum albumin. The result obtained by the difference spectrum method showed that 1 mol of each alpha-fetoprotein bound 1 mol of bilirubin at pH 8.3 and that the dissociation constants of the complexes of bilirubin with fetal alpha-fetoprotein and hepatoma-derived alpha-fetoprotein were 2.6 x 10(-7) and 5.0 x 10(-7) M, respectively. The Jacobsen enzymatic method using horseradish peroxidase gave the same values for molar binding ratios and similar dissociation constants, 7.1 x 10(-7) M for fetal alpha-fetoprotein and 7.4 x 10(-7) M for hepatoma-derived alpha-fetoprotein. These results indicate that alpha-fetoprotein may function as a carrier protein for bilirubin as has been shown for serum albumin.

Comparative chemical structures of human alpha-fetoproteins from fetal serum and from ascites fluid of a patient with hepatoma.

Human alpha-fetoproteins were purified from umbilical cord serum and from ascites fluid of a patient with hepatoma by affinity chromatography, and their chemical compositions and terminal sequences were compared. The amino acid compositions of these alpha-fetoproteins were similar and in good agreement with the values reported by other investigators. The COOH-terminal 5-amino acid sequence determined by carboxypeptidase digestion and the NH2-terminal 20-amino acid sequence determined by an automated sequence analyzer revealed that both alpha-fetoproteins had the same terminal sequences of amino acids. The sequence analysis showed that a part of each of the proteins lacked its NH2-terminal residues for one or three amino acids. A small difference in the carbohydrate composition of each alpha-fetoprotein was observed. It was concluded that alpha-fetoproteins from fetal serum and from ascites fluid of a patient with hepatoma had very similar structures.

Copper(II)-binding ability of human alpha-fetoprotein.

The copper(II)-binding ability of human alpha-fetoproteins, which were purified from umbilical cord serum and from ascites fluid of a hepatoma-bearing patient, was examined by equilibrium dialysis and gel filtration methods. The pH dependence of the copper(II)-binding ability of alpha-fetoprotein was quite similar to that of albumin. Alpha-fetoprotein bound 1 mol of copper(II) ion per mol of protein above pH 6.0 and 0.5 mol of copper(II) ion at pH 5.4, which is close to the pK value of the imidazole group of histidine. Photooxidation of alpha-fetoprotein in the presence of methylene blue resulted in the loss of the copper(II)-binding ability of the protein in parallel with the destruction of the histidyl residues. A synthetic amino-terminal undecapeptide of alpha-fetoprotein also bound copper(II) ion. These results indicate that the histidyl residue at the amino-terminal region of alpha-fetoprotein plays an important role in the copper(II)-binding ability of the protein.

Fucosylation of serum alpha-fetoprotein in patients with primary hepatocellular carcinoma.

alpha-Fetoprotein specimens were prepared from the sera of four patients with hepatocellular carcinoma. The lentil lectin-reactive and lectin-nonreactive variants of this glycoprotein were also prepared from the serum of one of the four patients by affinity chromatography with immobilized lectin. The correlation between the carbohydrate structure of these compounds and their reactivity in crossed immuno-affinoelectrophoresis with lentil lectin was studied by chemical analysis and affinity chromatography of the glycopeptides with lectin columns. It was found that the lentil lectin-reactive variant contained a carbohydrate chain of the fucosylated biantennary complex type. These data together with previous findings indicate that most of the patients with hepatocellular carcinoma have an elevated serum concentration of fucosylated alpha-fetoprotein.

Novel gene mutations in patients with left ventricular noncompaction or Barth syndrome.

BACKGROUND: Mutations in the gene G4.5 result in a wide spectrum of severe infantile cardiomyopathic phenotypes, including isolated left ventricular noncompaction (LVNC), as well as Barth syndrome (BTHS) with dilated cardiomyopathy (DCM). The purpose of this study was to investigate patients with LVNC or BTHS for mutations in G4.5 or other novel genes. METHODS AND RESULTS: DNA was isolated from 2 families and 3 individuals with isolated LVNC or LVNC with congenital heart disease (CHD), as well as 4 families with BTHS associated with LVNC or DCM, and screened for mutations by single-strand DNA conformation polymorphism analysis and DNA sequencing. In 1 family with LVNC and CHD, a C-->T mutation was identified at nucleotide 362 of alpha-dystrobrevin, changing a proline to leucine (P121L). Mutations in G4.5 were identified in 2 families with isolated LVNC: a missense mutation in exon 4 (C118R) in 1 and a splice donor mutation (IVS10+2T-->A) in intron 10 in the other. In a family with cardiomyopathies ranging from BTHS or fatal infantile cardiomyopathy to asymptomatic DCM, a splice acceptor mutation in exon 2 of G4.5 (398-2 A-->G) was identified, and a 1-bp deletion in exon 2 of G4.5, resulting in a stop codon after amino acid 41, was identified in a sporadic case of BTHS. CONCLUSIONS: These data demonstrate genetic heterogeneity in LVNC, with mutation of a novel gene, alpha-dystrobrevin, identified in LVNC associated with CHD. In addition, these results confirm that mutations in G4.5 result in a wide phenotypic spectrum of cardiomyopathies.

Automatic border detection identifies subclinical anthracycline cardiotoxicity in children with malignancy.

BACKGROUND: Anthracycline drugs for cancer therapy often cause functional myocardial impairment even in relatively low doses. We investigated the left ventricular function in asymptomatic anthracycline-treated children by automatic border detection (ABD) to assess its clinical usefulness for unmasking latent anthracycline-induced myocardial damage. METHODS AND RESULTS: Thirty-four children (0.7 to 17.6 years old) during or after anthracycline chemotherapy (26 to 1100 mg/m2) for malignancy (Chemo group) were studied, and 40 children (2.8 to 15.6 years old) without cardiac involvement served as normal control subjects (Control group). All patients underwent complete echocardiographic examination, including M-mode, Doppler, and ABD. Conventional echocardiography disclosed no difference between groups with regard to ejection fraction and the ratio of early to late transmitral flow velocity. In marked contrast, an investigation using ABD revealed that the Chemo group appeared to have some anthracycline-induced myocardial damage. In the apical 4-chamber view, peak filling rate in the Chemo group [2.3+/-0.4 end-diastolic area (EDA)/s] was significantly lower than that in the Control group (3.1+/-0.5 EDA/s) (P<0.0001), and time to peak filling rate in the Chemo group (106+/-31 ms) was clearly prolonged compared with that in the Control group (74+/-22 ms) (P<0.0001). CONCLUSIONS: Echocardiographic ABD may be a sensitive and useful noninvasive approach for evaluating subclinical anthracycline cardiotoxicity.

Clinical features of isolated noncompaction of the ventricular myocardium: long-term clinical course, hemodynamic properties, and genetic background.

OBJECTIVES: A nationwide survey was conducted to clarify the clinical features of isolated noncompaction of the ventricular myocardium (INVM) in Japanese children in comparison with features previously described in patients with INVM. BACKGROUND: Isolated noncompaction of the ventricular myocardium is a rare disorder characterized by an excessively prominent trabecular meshwork. It is accompanied by depressed ventricular function, systemic embolism and ventricular arrhythmia. METHODS: A questionnaire specifically designed for this study was sent to 150 hospitals in Japan where a pediatric cardiology division exists. RESULTS: Twenty-seven patients were diagnosed by two-dimensional echocardiography, their ages ranging from one week to 15 years at presentation, with follow-up lasting as long as 17 years. The gross anatomical appearance and the extension of noncompacted myocardium predominantly at the apex observed on two-dimensional echocardiograms were similar to observations reported previously. Dissimilarities included a greater number of asymptomatic patients at initial presentation, a longer clinical course with gradually depressed left ventricular function, no systemic embolism, and rare ventricular tachycardia in the Japanese children. Cardiac catheterization disclosed normal left ventricular end-diastolic volume and increased left ventricular end-diastolic pressure in most cases, consistent with restrictive hemodynamics. A higher incidence of Wolff-Parkinson-White syndrome was found in the children, whereas left bundle branch block was rarer than reported in adults. Familial recurrence was high (44%) and included many women. CONCLUSIONS: In Japanese children, INVM can be found by screening examinations at asymptomatic stage, and it might have a longer dinical course with gradually depressed left ventricular function and restrictive hemodynamics. The pattern of familial recurrence we observed implies that INVM is a distinctive clinical entity with a heterogeneous genetic background.

Rug shampoo and Kawasaki disease.

Following personal interviews with 63 families of children with Kawasaki disease and with 63 control families with children paired for race, sex, and age, no epidemiologic differences were seen except for use of rug shampoo within 1 month of onset in 16 episodes in 15 children with Kawasaki disease in 14 families (24% of children, 22% of families) compared with two families of control children (3%, P less than .005). Explanations for this strong association of recent rug shampoo and Kawasaki disease include the question of recall bias as well as the possibility that an agent in the shampooing process does cause or does contribute to illness.

Coronary artery involvement in Kawasaki syndrome in Manhattan, New York: risk factors and role of aspirin.

Since January 1980, 110 children having 113 attacks of Kawasaki syndrome were studied. Age at onset was 7 weeks to 12 years (mean 3 6/12 years, median 2 9/12 years); 77% were younger than 5 years of age; the male to female ratio was 1.8; racial distribution was 52% white, 19% black, 14% Hispanic, and 16% Asian. Protocol of management consisted of high-dose aspirin (100 mg/kg/d) until afebrile, and then 81 mg every day until free of coronary aneurysm. Two-dimensional echocardiograms were done weekly during the acute stage, at 2 and 6 months after onset, and yearly if a coronary abnormality was detected. At 1 month, 51 coronary arterial abnormalities were present in 25 patients. Risk factors for a coronary abnormality were duration of fever greater than or equal to 2 weeks, level of platelet count, marked elevation of ESR, and age younger than 5 years. No statistically significant difference in incidence of aneurysms was detected between patients on high-dose aspirin and those on medium-or low-dose aspirin.

Epidemiologic aspects of Kawasaki disease in a Manhattan hospital.

Epidemiologic and clinical features of Kawasaki disease in 106 patients seen between 1980 and 1986 at The New York Hospital in midtown Manhattan were compared with those in large series from the United States, Canada, and Japan. Dissimilarities in our Kawasaki disease experience included ethnic heterogeneity of our patients (50% white, 18% black, 16% Hispanic, and 16% Oriental) and, in comparison with the Japanese experience, an older mean age (3 1/2 vs 1 1/2 years) with fewer children less than 2 years of age (32% vs 50% to 60%). In comparison with the general population of the geographic urban and suburban referral area for our hospital and in comparison with our general pediatric population, Oriental children with Kawasaki disease were overrepresented (16% vs 2%). More families of children with Kawasaki disease were members of the upper and middle class (73%) than were the population seen in general pediatrics (31.7%) at our hospital. Personal interviews with 63 families of children with Kawasaki disease and 63 control families with children paired for ethnic group, sex, and age revealed no epidemiologic differences except for use of rug shampoo within 1 month of onset in 16 episodes in 15 children with Kawasaki disease in 14 families (22% of families) compared with two families of control children (3%) (P less than .001).

Additive effect of beraprost on pulmonary vasodilation by inhaled nitric oxide in children with pulmonary hypertension.

Combined administration of inhaled nitric oxide and beraprost sodium resulted in a more intense decrease in pulmonary vascular resistance than nitric oxide given alone (mean -33% vs -45%, p <0.05), without serious systemic hypotension. Combined therapy with nitric oxide and beraprost sodium is highly desirable in treating primary and secondary pulmonary hypertension in children.

Treatment of hepatocellular carcinoma utilizing lymphokine-activated killer cells and interleukin-2.

This paper is a report on adoptive immunotherapy involving consecutive injections of recombinant interleukin-2 and lymphokine-activated killer (LAK) cells in the treatment of hepatocellular carcinoma. Peripheral blood lymphocytes, obtained by leukopheresis, acted as the activated killer cells with a co culture of recombinant interleukin-2 in the culture system. After 4 days, the activated killer cells were returned into the patients' bodies intra-arterially and intravenously. No complete remissions or partial remissions have resulted, although five of the seven patients showed a significant decrease in their serum alpha-fetoprotein levels after treatment. In addition, one case showed a patent portal truncus while another indicate the appearance of central necrosis on the computerized tomograph scan. Although the period of observation was short, there were no recurrences after the combination therapy of tumor resection and LAK adoptive immunotherapy. It might be difficult to treat hepatocellular carcinoma with adoptive immunotherapy alone, but there is some possibility of conducting therapy for hepatocellular carcinoma after removing the majority of the tumor cells by surgical resection and transcatheter arterial embolization therapy. This conclusion indicates, at least theoretically, that adoptive immunotherapy will be suitable in the treatment of hepatocellular carcinoma as one of the combination therapies with the two major forms of treatment mentioned above.

Dipyridamole stress ultrasonic myocardial tissue characterization in patients with Kawasaki disease.

Dipyridamole stress integrated backscatter (IBS) was used for evaluation of myocardial ischemia or damage in 31 children with coronary artery lesions caused by Kawasaki disease, in comparison with thallium-201 myocardial imaging. All patients underwent echocardiography at rest and after dipyridamole stress at the anterior interventricular septum, posterior wall (PW), and inferior wall (INF). At rest, no significant difference was seen in cyclic variation (CV) of IBS in the regions with normal or abnormal distribution on Tl-201 imaging. But in the regions showing abnormal distribution after stress, CV decreased significantly. A delayed study after stress showed the recovery of CV to the level at rest in all patients. Sensitivity of abnormal cyclic variation integrated backscatter was 75% in the PW and 91% in the INF, and specificity was 91% in the PW and 90% in the INF, compared with the results of thallium-201 imaging. Dipyridamole stress IBS can provide sensitive detection of myocardial ischemia or damage in Kawasaki disease.

Clinical evaluation of transcatheter arterial embolization and one-shot chemotherapy in hepatocellular carcinoma.

The therapeutic effect and the prognosis of transcatheter arterial embolization (TAE) and one-shot chemotherapy were studied in 90 cases of unresectable hepatocellular carcinoma (HCC). A therapeutic effect, which was assessed by the serum concentration of alpha-fetoprotein, angiography, computed tomography and ultrasonography, was seen in 33 (83%) out of 40 cases treated with TAE, and in 16 (32%) out of 50 cases treated with one-shot chemotherapy. In the cases that received TAE, the cumulative percentage survival rates at 6 months, 12 months, and 24 months were 75%, 48% and 20%, respectively. In contrast, the survival rates in the cases that received one-shot chemotherapy were 30%, 10% and 2%, respectively. In addition, the prognosis on the basis of the degree of tumor invasion of the portal vein was studied. In cases with the same degree of tumor invasion, the survival rate of the cases treated with TAE was significantly higher than that of those treated with one-shot chemo-therapy, except for those cases with tumor invasion of the main portal vein. These results show that TAE should be performed as the therapy of first choice in unresectable cases of HCC.

Treatment of HBeAg-positive chronic hepatitis with a streptococcal preparation (OK-432).

Forty-two patients with hepatitis Be antigen (HBeAg)-positive chronic hepatitis were treated by intramuscular injections with OK-432, an immunopotentiator possessing interferon-inducing activity. They were monitored with serial measurements of virological parameters to evaluate therapeutic effectiveness, and compared with a group of seventy-five untreated patients (natural course group). In the group receiving OK-432 therapy, twenty-seven patients (64.3% of the forty-two patients) became negative for HBeAg in an average observation period of 20.1 months. Of these, fourteen patients (33.3% of the forty-two patients) underwent seroconversion from HBeAg to anti-HBe antibody (anti-HBe). In the natural course group, twenty-three patients (30.7% of the seventy-five patients) lost HBeAg reactivity in a mean follow-up period of 32.3 months, and thirteen patients (17.3% of the seventy-five patients) became seroconverted. Thus, the drug group showed significantly higher percentages of patients with disappearance of HBeAg and seroconversion, notwithstanding the shorter duration of the follow-up. Young males and females, females generally, or patients with high serum GPT levels were more likely to respond to the therapy. The serum GPT level tended to stabilize more in patients receiving OK-432.

Clinical study of recombinant hepatitis B vaccine.

The efficacy and safety of a recombinant yeast-derived hepatitis B vaccine were evaluated in 209 subjects after three administrations at 0, 4 and 20 weeks. Subjects were divided into four groups given 5 micrograms vaccine subcutaneously, 10 micrograms subcutaneously, 10 micrograms intramuscularly and 20 micrograms subcutaneously to define the effective dose and to compare the effect of administration. Seroconversion of the antibody to hepatitis B surface antigen after the third vaccination reached 96.6% in the group given 5 micrograms vaccine subcutaneously and 100% in the other groups. The final geometric mean antibody titres were 700 IU/l in subjects given 5 micrograms subcutaneously, 2004 IU/l in those given 10 micrograms subcutaneously, 4674 IU/l in those given 10 micrograms intramuscularly and 3342 IU/l in those given 20 micrograms subcutaneously. In the groups given 10 micrograms, the early seroconversion rate of the antibody to hepatitis B surface antigen and the geometric mean antibody titres after the third vaccination were significantly higher in subjects administered intramuscularly than subcutaneously (P less than 0.05). No major adverse effects were observed and minor reactions were the same as, or less than, those reported for the plasma-derived vaccine. Before and after administration, no significant fluctuation in the yeast antibody titre was observed. These results demonstrate the efficacy and safety of the yeast-derived vaccine, and show that 10 micrograms was the effective dose.

Therapeutic effects of tiopronin on chronic hepatitis: a double-blind clinical study.

The efficacy of tiopronin (2-mercaptopropionyl glycine) 600 mg daily in the treatment of chronic active or chronic persistent hepatitis has been assessed in a double-blind controlled clinical trial of 12 weeks involving 165 Japanese patients with histologically proven disease. Treatment with the drug was associated with a significant improvement in abnormalities of serum transaminase and gamma glutamyl transpeptidase, with reversion towards baseline values on stopping the drug. Improvement was independent of the histological classification of the disease, or HBsAg status. The drug was well tolerated with few side-effects. The results of this short-term study indicate that tiopronin may be of value in the treatment of chronic hepatitis.

Total anomalous pulmonary venous connection with bronchogenic cyst in neonatal period.

Here, we report a case of a two-day-old neonate with total anomalous pulmonary venous connection to the innominate vein and a bronchogenic cyst arising from the trachea. Antenatal echocardiography had delineated both cardiac and extracardiac lesions, and a repeated examination on the day of birth disclosed progressive enlargement in the cyst in a manner so as to obstruct the innominate vein. On the second day of life, the patient underwent complete correction of the cardiac lesion and total excision of the cyst. The patient recovered uneventfully and was discharged on the thirteenth postoperative day.