Anti-tumor polysaccharide from the mycelium of liquid-cultured Agaricus blazei mill.

Anti-tumor active polysaccharide against Sarcoma 180 was isolated by DEAE-Sepharose CL-6B and Sepharose 4B column chromatography from the hot-water soluble fraction of the mycelium of liquid-cultured Agaricus blazei mill. This polysaccharide did not react with antibodies of anti-tumor polysaccharides such as lentinan, gliforan, and FIII-2-b which is one of anti-tumor polysaccharides from Agaricus blazei. Moreover, the analyses of 13C-NMR and GC-MS suggested that this polysaccharide was preliminarily glucomannan with a main chain of beta-1,2-linked D-mannopyranosyl residues and beta-D-glucopyranosyl-3-O-beta-D-glucopyranosyl residues as a side chain. This polysaccharide was completely different from the anti-tumor polysaccharide from fruiting body of Agaricus blazei, beta-1,6-glucan.

Antitumor effects of a new polysaccharide-protein complex (ATOM) prepared from Agaricus blazei (Iwade strain 101) "Himematsutake" and its mechanisms in tumor-bearing mice.

The antitumor activity of the i.p. or p.o. administration of polysaccharide-protein complex, ATOM (antitumor organic substance Mie) prepared from cultured mycelia of Agaricus blazei (Iwade strain 101) "Himematsutake" examined against four kinds of established mouse tumors. ATOM was highly effective at the doses of 10 and 20 mg/kg/day x 10 on subcutaneously implanted Sarcoma 180 in mice, and was also active against Ehrlich ascites carcinoma, Shionogi carcinoma 42 and Meth A fibrosarcoma at doses of 50 and 100 mg/kg/day x 10. ATOM has no direct cytotoxic action on tumor cells in vitro. Thus the tumor growth-inhibitory effect of ATOM is apparently due to immunological host-mediated mechanisms. The number of peritoneal macrophages, the phagocytosis of polystyrene latex beads and the proportion of the third component of complement (C3)-positive fluorescent cells were increased in the mice treated with ATOM. These results suggest that the macrophage activa-tion and alterations of the C3 are necessary for the induction of an antitumor effect of ATOM.

Life-threatening cardiac involvement throughout life in a case of Costello syndrome.

Costello syndrome is characterized by poor postnatal growth, mental retardation, curly hair, coarse face, loose skin of the hands and feet, and nasal papillomata. Patients with Costello syndrome have a high incidence of cardiac involvement, such as arrhythmias, hypertrophic cardiomyopathy, or congenital anomalies. The importance of cardiac involvement in Costello syndrome has not been strongly emphasized thus far, although arrhythmia and hypertrophic cardiomyopathy are both serious forms of cardiac involvement. We report the case of a Japanese girl with Costello syndrome, who experienced life-threatening cardiac involvement throughout her life.

Measurement and validity of the ventilatory threshold in patients with congenital heart disease.

The purpose of the present study was to measure oxygen uptake (VO2) at the ventilatory threshold (VT) in patients with congenital heart disease using a progressive exercise protocol on a treadmill and to evaluate the validity and feasibility of this procedure. Eight control subjects and seventeen patients performed a maximal exercise test with breath-by-breath measurement of ventilation and gas exchange variables. VT(VE) was determined by the change in the ventilatory equivalent for VO2 and carbon dioxide output, VT(V-sl) by the V-slope method, and the lactate threshold (LT) by the change in blood lactate concentration; these parameters were determined in 100%, 88%, and 96% of subjects, respectively. The interobserver error among three evaluators was not significant, and LT was correlated with each VT (r = 0.97, 0.92; p = 0.0001) and with peak VO2 (r = 0.91; p = 0.0001). The VTs were correlated with each other when expressed as milliliter per minute and milliliters per kilogram per minute. It was concluded that a progressive exercise protocol on a treadmill was a feasible procedure for determining the VTs in most individuals and that VTs were valid, useful parameters for evaluating submaximal exercise tolerance in patients with congenital heart disease.

Abnormal B cell response of protein kinase C in some common variable immunodeficiency.

The localization of protein kinase C (PKC) was investigated in B cells or B cell lines derived from patients with common variable immunodeficiency (CVI) and healthy controls. Stimulation of the healthy control B cells by phorbol ester or anti-mu induced PKC activation and translocation to the plasma membrane after 10 min. In contrast, in the CVI-B cells, no apparent PKC translocation was induced by either phorbol ester or anti-mu stimulation. These results suggest that the pathogenesis in some CVI patients might involve defects in the pathway of PKC activation.

Abnormal response to exercise, face immersion, and isoproterenol in children with the long QT syndrome.

The present study was performed to observe the change of QT interval by sympathetic stimulations in patients with the long Qt syndrome (LQTS). The study group consisted of 6 children with LQTS and 6 healthy children without QT prolongation. All LQTS patients had syncopal episodes. The QTc and delta QTc% ([QTc interval after examination-QTc interval at rest]/ QTc interval at rest x 100) by treadmill testing, face immersion, and isoproterenol were examined. One minute after peak exercise of treadmill testing, the changes in the QTc interval were not significant in either group, but delta QTc% was larger in the LQTS group than in the control group (+ 11.0 +/- 12.1% vs -2.6 +/- 3.2%; P = 0.02). The QTc interval at the shortest RR interval during face immersion was prolonged in the LQTS group (0.47 +/- 0.01 s to 0.51 +/- 0.04 s; P = 0.02), but there were no significant changes in the control group (0.40 +/- 0.03 s to 0.41 +/- 0.03 s; P = NS). delta QTc% was larger in the LQTS group than in the control group (+ 10.0 +/- 7.3% vs +1.1 +/- 5.5%; P = 0.04). In the LQTS group, the RR interval was shortened (P = 0.009) and QTc interval was prolonged (P = 0.0008) after isoproterenol infusion. These sympathetic stimulations amplified the TU abnormality in the LQTS group. By observing the TU changes caused by face immersion, we hoped to find a possible new method with which to diagnose LQTS. The combination of these examinations may be helpful in screening the borderline cases of TU abnormalities.

Effects of verapamil and propranolol on early afterdepolarizations and ventricular arrhythmias induced by epinephrine in congenital long QT syndrome.

OBJECTIVES: This study used monophasic action potentials to investigate the effects of verapamil and propranolol on epinephrine-induced repolarization abnormalities in congenital long QT syndrome. BACKGROUND: Early afterdepolarizations have been suggested to play a significant role in QT prolongation and ventricular arrhythmias in congenital long QT syndrome. Calcium channel blocking as well as beta-adrenergic blocking agents are reported to be effective in the management of this syndrome. METHODS: Monophasic action potentials from 2 to 4 sites were recorded simultaneously in eight patients with the long QT syndrome (22 sites) and in eight control patients (23 sites) and were obtained during constant atrial pacing 1) before epinephrine infusion; 2) during epinephrine infusion (0.1 microgram/kg body weight min); 3) after verapamil injection (0.1 mg/kg) during epinephrine infusion; and 4) after both propranolol (0.1 mg/kg) and verapamil injections. RESULTS: Early afterdepolarizations were recorded in two of the eight patients (2 of 22 sites) during the control state. During epinephrine infusion, early afterdepolarizations were recorded in six patients (six sites), and ventricular premature complexes were induced in three and torsade de pointes in one. Epinephrine prolonged 90% monophasic action potential duration from 348 +/- 48 (mean +/- SD) to 381 +/- 49 ms (22 sites, p < 0.0005) and increased the dispersion of action potential duration (difference between the longest and shortest action potential duration) from 36 +/- 20 to 64 +/- 34 ms (p < 0.005). Verapamil eliminated (two sites) or reduced (four sites) early afterdepolarizations and abolished ventricular premature complexes in two of the three patients as well as suppressing torsade de pointes. Verapamil shortened the action potential duration to 355 +/- 28 ms (p < 0.01 vs. epinephrine) and decreased the dispersion to 44 +/- 19 ms (p < 0.05 vs. epinephrine). Propranolol further eliminated (two sites) or reduced (two sites) early after depolarizations, abolished ventricular premature complexes in the remaining one patient and further shortened the action potential duration to 337 +/- 32 ms (p = 0.09 vs. verapamil). In the control patients, none of the early afterdepolarizations, ventricular arrhythmias or marked prolongations of action potential duration were induced by epinephrine, and neither verapamil nor propranolol changed repolarization variables. CONCLUSIONS: These results indicate that both verapamil and propranolol can improve repolarization abnormalities induced by epinephrine in congenital long QT syndrome.

Provocative testing and drug response in a patient with the long QT syndrome.

A girl of 14 with the long QT syndrome (LQTS) and torsades de pointes is reported. Isoprenaline or adrenaline infusions induced torsades de pointes and inversion of the TU wave. Changes in the TU wave during isoprenaline infusion were used to select effective drugs to treat this patient. A beta blocker and calcium channel blocker were selected and the patient had no episodes of syncope for two years. This electrocardiographically guided method may be useful for selecting effective drugs in patients with the LQTS.

Fractionation and Antitumor Activity of Polysaccharides from Grifola frondosa Mycelium.

We developed a method for the fractionation and purification of antitumor polysaccharides, considered to be a type of immuno-potentiator or BRM (biological response modifier), from the mycelium of liquid cultured Grifola frondosa. The active polysaccharide fractions that showed higher antitumor activity were considered to be heteroglycans or their protein complexes as follows, in water-soluble fractions: FI0-a-α: fucoga-lactomannan-protein complex; FI0-a-ß: mannogalactofucan; FA-1: galactoglucomannofucan-protein complex; FA-2-b-α: glucogalactomannan-protein complex; in water-insoluble fractions: FIII-1-a: mannofucoglucoxylan; FIII-1-b: mannoglucofucoxylan-protein complex; FIII-2-a: mannofucoglucoxylan-protein complex; FIII-2-b: glucomannofucoxylan-protein complex.

[Sequential changes in plasma lipoprotein(a) as acute phase protein after myocardial infarction and surgery group].

The plasma lipoprotein(a), apo AI, apo B, and acute phase proteins, were studied in 21 patients with myocardial infarction and in 11 patients after surgery. In the both groups, C-reactive protein showed a rapid increase, and alpha 1 acid glycoprotein, alpha 1 antitrypsin and lipoprotein(a) followed by a moderate increase and restored to normal values after one month. Lipoprotein(a) increased to a maximum on day 11 in the myocardial infarction group, and on day 8 in the surgery group. Only slight changes in apolipoprotein AI and B were noted. We speculate that lipoprotein(a) is an acute phase protein that plays an important roles in recovery from trauma. Recently it was reported that the amino acid sequence of apolipoprotein(a) is partly identical to that of plasminogen. This sequence suggests that lipoprotein(a) and plasminogen are related immunochemically. We examined by immunoblotting technique whether our antibody for lipoprotein(a) is influenced by a plasminogen in plasma. By enzyme-linked immunosorbent assay, it was found that the purity of plasminogen does not influence determination of lipoprotein(a).

Transient changes of serum lipoprotein(a) as an acute phase protein.

Serum lipoprotein(a) (Lp(a)) was serially determined after acute attacks of myocardial infarction and after surgical operations. Acute phase proteins, such as C-reactive protein, alpha 1-acid glycoprotein, alpha 1-antitrypsin and haptoglobin, increased rapidly and markedly after the episodes. Initial values of serum Lp(a) concentrations were almost the same in both groups. Increases in serum Lp(a) levels were also observed during the first few days, with a return to the initial levels after more than 1 month. The periods for reaching maximal levels of acute phase proteins were similar in both groups of patients. On the contrary, the period required for Lp(a) to reach the maximal level in the myocardial infarction group was significantly longer than in the post-operative group. The present study suggests that Lp(a) has the characteristics of an acute phase reactant and may play an important role in recovery from tissue damage.

Lipoprotein(a) in nonhuman primates. Presence and characteristics of Lp(a) immunoreactive materials using anti-human Lp(a) serum.

Lipoprotein(a) (Lp(a] immunoreactive materials were examined in serum samples from 77 nonhuman primates of 24 species by Ouchterlony's double diffusion procedure and an enzyme-linked immunosorbent assay (ELISA) using rabbit antisera to human Lp(a). The precipitates obtained with sera from orang-utan and chimpanzee formed reactions of complete identity with the Lp(a) precipitate with human serum. When sera from Old World monkeys and human subjects were tested in wells next to each other, spurs developed between the 2 precipitates, indicating that Lp(a)-like lipoproteins in Old World monkeys have partial identity with human Lp(a). Lp(a) immunoreactive materials were identified in association with lipids by means of fat staining of the precipitates. On the other hand, reactants which could be precipitated with anti-human Lp(a) sera were not detectable in prosimians and New World monkeys. These results suggest that serum Lp(a)-like lipoprotein is phylogenetically acquired in Old World monkeys. However, the possibility that the structures of serum Lp(a)-like lipoproteins in prosimians and New World monkeys are too different to react with anti-human Lp(a) sera cannot be ruled out.

Enzyme-linked immunosorbent assay of lipoprotein(a) in serum and cord blood.

We have developed a new sensitive method for quantifying lipoprotein(a) (Lp(a] in human serum, using a 'sandwich' type noncompetitive enzyme-linked immunosorbent assay (ELISA). The solid-phase used was a polystyrene plate. The anti-Lp(a) antibody-enzyme conjugate was labelled by linking Fab' fragments to peroxidase (EC 1.11.1.7) by the maleimide method. The minimum detectable concentration was 0.5 ng/well. Routinely, the assay was carried out with 1,000-fold diluted serum, and Lp(a) was quantified between 4.0 and 500 mg/l. Within-run coefficients of variation (CVs) ranged from 3.5% to 10.4% and between-run CVs from 5.0% to 11.1%. Results by the ELISA were in good agreement with those by radial immunodiffusion (r = 0.955). The distribution of Lp(a) in serum from 820 healthy donors was highly skewed: mean 141.1 mg/l, medium 97.9 mg/l. In cord blood, the mean and median were 15.6 and 9.8 mg/l, respectively. This ELISA for Lp(a) has the advantages of being highly sensitive and specific, simple to perform, and does not use radioisotopes.

Two cases of systemic lupus erythematosus associated with hyperthyroidism.

We have experienced two cases (Case 1: 21-year-old female, Case 2: 26-year-old female) of systemic lupus erythematosus (SLE) associated with hyperthyroidism. Case 1 had been treated with methimazole (MMI) and betamethasone for approximately two years. Although thyroid function improved with the treatment, laboratory data of SLE deteriorated. She was successfully treated with betamethasone alone. Case 2, who had severe side effect (severe hemorrhage due to gastric ulcer) during prednisolone treatment for SLE, was found to have an additional hyperthyroidism. She was treated with intermittent prednisolone administration alone. Physical findings as well as laboratory data of both SLE and hyperthyroidism improved by the therapy.

Dynamics of plasma lipoproteins and lipids during double filtration plasmapheresis (DEP).

Plasma lipids and lipoproteins obtained from different places of the circuit of double filtration plasmapheresis (DFP) were measured and effect of DFP therapy on removal of them was examined. When 2A was used as a second filter, 69.8%, 52.4%, 63.0%, 58.0%, 60.8%, 59.2% and 63.9%, respectively of beta-lipoprotein, cholesterol, triglyceride, high density lipoproteins (HDL), phospholipids, free cholesterol and lipoprotein (a) (Lp(a)) were removed from the patient's plasma. When 4A was used as a second filter, 69.0%, 56.8%, 53.2%, 45.4%, 56.0%, 50.9% and 51.7%, respectively of beta-lipoprotein, cholesterol, triglyceride, HDL, phospholipids, free cholesterol and Lp(a) were removed from the patient's plasma. In contrast, concentrations of free fatty acids (FFA) after DFP therapy using filter 4A and 2A increased to 222.8% and 256.4%, respectively. Thus, it was shown that except for FFA, DFP therapy using either 2A or 4A as a second filter is effective in reducing concentrations of plasma lipid and lipoproteins.