Measurement of plasma brain natriuretic peptide level as a guide for cardiac overload.

OBJECTIVES: We examined whether measurement of the plasma BNP concentrations might be useful for the early diagnosis of the existence and severity of disease in patients with heart disease in daily clinical practice. METHODS AND RESULTS: The plasma BNP and ANP concentrations in 415 patients with heart disease and hypertension and 65 control subjects were measured. Patients with heart disease had higher plasma BNP and ANP concentrations than did those with hypertension or control subjects. Among the etiology of cardiac diseases, specifically dilated cardiomyopathy and hypertrophic cardiomyopathy, was associated with the highest plasma BNP concentrations, whereas dilated cardiomyopathy was associated with the highest plasma ANP concentrations. Plasma BNP concentrations showed an increase as the severity of the heart disease, as graded according to the NYHA classification of cardiac function, increased. In both patients with heart disease and hypertension, the plasma BNP values were higher in those who had abnormalities in their echocardiogram and electrocardiogram as compared to those without any abnormalities. The plasma BNP levels also showed a significant correlation with left ventricular wall thickness and left ventricular mass. On the other hand, the plasma ANP levels showed significant correlations with left ventricular dimension. Receiver operative characteristic analysis revealed that plasma BNP levels showed substantially high sensitivity and specificity to detect the existence of heart diseases. CONCLUSION: Measurements of the plasma BNP concentrations is useful to detect the existence of the diseases, and abnormalities of left ventricular function and hypertrophy in patients with heart disease in daily clinical practice.

Enhancement by retinoid of hemin-induced differentiation of human leukemia K562 cell line.

The effect of retinoid on human leukemia K562 cell differentiation induced by hemin was examined. Retinoids (retinoic acid and synthetic retinoids [Am80 and Ch55]) dose-dependently enhanced hemin-induced erythroid differentiation of K562 cells, though these retinoids themselves did not induce the differentiation. Under optimal conditions, these retinoids caused a doubling of the population of hemin-induced differentiated cells. In addition, co-treatment of cells with hemin and retinoid led to longer maintenance of the differentiated state after the removal of hemin, which might imply acquisition of irreversibility of hemin-induced differentiation. These results suggest that the combination of retinoids with other differentiation inducers might be useful for leukemia therapy in cases where the leukemic cells are poorly responsive or unresponsive to retinoids, alone.

Expression of opioid-binding cell adhesion molecule (OBCAM) and neurotrimin (NTM) in E. coli and their reactivity with monoclonal anti-OBCAM antibody.

Opioid-binding cell adhesion molecule (OBCAM), neurotrimin (NTM) and limbic system-associated membrane protein (LAMP) are homologous and are the members of the IgLON family which is a subfamily within the immunoglobulin superfamily. We cloned the cDNAs for OBCAM and NTM, prepared recombinant proteins, and examined the reactivity of the previously prepared monocolonal anti-OBCAM antibody, OBC53, with the recombinant proteins by immunoblotting. These experiments revealed that OBC53 recognizes OBCAM about 1000 times as efficiently as NTM. Moreover, the NTM and LAMP peptides which have sequences homologous to the OBCAM peptide used for the preparation of OBC53 were 150 times less reactive to OBC53. Thus, the OBC53 antibody is a useful tool for specifically detecting OBCAM in immunochemical experiments.

Animal models for X-linked sideroblastic anemia.

Erythroid 5-aminolevulinate synthase (ALAS-E) catalyzes the first step of heme biosynthesis in erythroid cells. Several lines of evidence suggest that the expression of ALAS-E is important for the process of erythroid differentiation, which requires a large amount of heme for hemoglobin production. Mutation of human ALAS-E causes the disorder X-linked sideroblastic anemia (XLSA). More than 25 unrelated ALAS-E mutations in XLSA patients have been reported. Most XLSA cases are of the pyridoxine-responsive type, but molecular diagnosis of 1 pyridoxine-refractory type XLSA has also been reported. To examine the roles heme plays during hematopoiesis and to create animal models of XLSA, we disrupted the mouse ALAS-E gene. A chemically induced zebrafish mutant (sau) that lacks ALAS-E has also been isolated. Analysis of these ALAS-E mutants unequivocally demonstrated that ALAS-E is the principal isozyme contributing to erythroid heme biosynthesis In ALAS-E-null mutant mouse embryos, erythroid differentiation was arrested, and an abnormal hematopoietic cell fraction emerged that accumulated a large amount of iron diffusely in the cytoplasm. This accumulation of iron was in contrast to that in XLSA patients, as typical ring sideroblasts accumulated iron primarily in mitochondria. These observations suggest that the mode of iron accumulation caused by the lack of ALAS-E is different in primitive and definitive erythroid cells. Thus ALAS-E, and hence heme supply, is necessary for erythroid cell differentiation and iron metabolism.

Localization of opioid-binding cell adhesion molecule (OBCAM) in adult rat brain.

We investigated the tissue distribution and brain localization of opioid-binding cell adhesion molecule (OBCAM) in the adult rats by immunoblotting and immunohistochemistry using a monoclonal anti-OBCAM peptide antibody that is specific for OBCAM. OBCAM was preferentially expressed in the central nervous system (CNS) and at a very low level in the spleen. Within the brain, OBCAM was distributed in almost all the gray matter, but little or no immunoreactive OBCAM was found in the white matter. Morphologically, the distribution pattern of OBCAM immunoreactivity was very similar to that of synaptophysin, suggesting a role in the synaptic machinery.

Developmental expression of opioid-binding cell adhesion molecule (OBCAM) in rat brain.

Opioid-binding cell adhesion molecule (OBCAM), a neuron-specific protein, consists of three immunoglobulin (Ig)-like domains anchored to the membrane through a glycosylphosphatidylinositol (GPI)-tail. OBCAM has been presumed to play a role as a cell adhesion/recognition molecule, but its function has not been fully elucidated. We investigated the developmental expression of OBCAM in rat brain by using a monoclonal anti-OBCAM peptide antibody (OBC53). OBCAM was clearly detectable on embryonic day 16 (E16) as assessed by immunoblotting. The expression level increased by the second postnatal week and was maintained at a constant level until week 17. During the early developmental period OBCAM was found to be expressed on postmitotic neurons and to be strongly expressed in at the fiber tracts containing expanding axons, in contrast to the adult brain, in which OBCAM is principally expressed in the gray matter. These findings suggest that the function of OBCAM involves axonal outgrowth.

[Chemical control of cell differentiation of human myeloleukemia K562 cell line].

Human myeloid leukemia K562 cells can be induced to differentiate to mature cells bidirectionary, i.e., hemin induces erythroid differentiation, while 12-O-tetradecanoylphorbol 13-acetate (TPA) induces differentiation to monocytes. The differentiation-inducing activity of various hemin-related compounds suggested certain structural requirements for the activity: 1) the iron moiety of hemin is not essential, and 2) the propionic acid side chains of hemin play an important role in the differentiation and induction. In addition, we have examined the influence of some bioresponse-modifying factors on hemin/protoporphyrin IX-induced differentiation of K562 cell line. Retinoids and tubulin-disruptors, themselves did not induce differentiation, enhanced hemin/protoporphyrin IX-induced differentiation of K562 cells. We also examined the possible involvement of peripheral-type benzodiazepine receptor (PBR) in hemin/protoporphyrin IX-induced differentiation on K562 cell lines. The PBR specific ligands modified hemin-induced differentiation. These results suggest a requirement for retinoids (or retinoids-like cofactors) for hemin/protoporphyrin IX-induced differentiation of K562 cells and the involvement of PBR in erythroid differentiation of K562 cell line. Further we showed that TPA suppresses hemin-induced erythroid differentiation of K562 cells, while retinoids augment it. TPA is a potent inducer of heme oxygenase (HO), which catabolizes heme to biliverdin. An HO inhibitor, tin protoporphyrin (SnPP), suppresses TPA-induced K562 cell differentiation to monocytes. It was also found that cotreatment of K562 cells with SnPP and TPA induces erythroid differentiation of K562 cells, though SnPP alone or TPA alone does not induce erythroid differentiation, suggesting a role of HO in the directional switch of differentiation.

[Neurogenic pulmonary edema following severe head injury--case report (author's transl)].

A case of severe head injury associated with fulminant pulmonary edema considered as neurogenic which developed within short time after the injury was presented. A five-year-old boy who had no previous history of cardiopulmonary disease was struck on his right frontal region by car accident at 15.30 PM on July 5 of 1979. Immediately after the impact he lost his consciousness and subsequently transferred to a local hospital where bilateral dilated pupil and flaccid paralysis of the limbs were noted. On transmission of the patient to Omuta City Hospital 30 minutes after the injury, massive foamy fluid was discharged from the tracheal tube. On admission, he was comatous, with bilateral dilated and fixed pupils and flaccid paralysis of the limbs. There was no retinal bleeding. He showed ataxic respiration with severe stridor and massive discharge of foamy fluid pinkish in colour from the trachea characteristic in pulmonary edema was significant. Chest x-ray film demonstrated perihilar densities suggesting pulmonary edema. CT scan showed extremely small ventricle on both sides without manifestations of intracranial hematomas or cerebral contusion. With an intensive medical treatments including corticosteroids, alkalizing agents and alpha-blocker were administered under controlled respiration, the discharge of edema fluid was gradually decreased and the findings on blood gases were also improved. However neurological signs were aggravated and he died 8 hours after the injury. Central venous pressure was maintained at the level between 8 to 10 cm. From these clinical findings the pulmonary edema was concluded as neurogenic. Direct or indirect injury to the hypothalamic efferent pathway at the level of lower brain stem seemed to be important as the cause of neurogenic pulmonary edema in this case. The possible pathophysiology of neurogenic pulmonary edema associated with brain stem injury and intracranial hypertension was discussed with other related literature.

[Measurement of anti-HTLV-I antibody titer in cerebrospinal fluid].

Anti HTLV-I antibody was measured in 590 cerebrospinal fluids (CSFs) employing a gelatin particle agglutination (PA) method. Anti HTLV-I antibody was detected in the CSFs from 24 patients (4.1%). The serum from the 24 patients also showed positive results. Out of the 24 patients, 11 cases (45.8%) had HAMs. Of the remaining patients 3 had ATLs with meningeal infiltration. Only 11 cases out of 24 patients were positive for anti HTLV-I antibodies when an immunofluorescence (IF) method was employed. This discrepancy of the results obtained by two methods may be explained by the presence of the patients whose CSFs contain anti HTLV-I antibodies of low titer. It is concluded that PA method is both sensitive and specific for the detection of anti HTLV-I antibodies in CSFs. Further, PA method is more convenient than IF method. So, it will be a very useful tool in the diagnosis of HTLV-I related neurological disorders and asymptomatic carriers.

[Comparative studies on the pharmacokinetics between THP and adriamycin in the same patients].

The pharmacokinetic properties of THP and ADM were comparatively studied in the same patients with various cancers. The concentration of ADM in either plasma or blood cells was higher than that of THP from 5 minutes to 24 hours after administration. The metabolites of ADM such as aglycones were detected in plasma until 3 hours after administration, but these were never detected in blood cells. By contrast, the metabolites of THP were detected until 24 hours after administration. These results suggested that THP was metabolized in tissues and excreted into urine and that the rate of metabolism and excretion of THP was faster than that of ADM. Distribution volumes (V1, V2, and V3) of THP were larger than those of ADM. The above results strongly suggested that THP would be easily transferred into the tissues, but that in the case of slower transferring tissues with lower K13 values, ADM rather than THP would be easily transferred to the tissues. These pharmacokinetic properties suggested that the toxicity of THP might be diminished compared with that of ADM.

[Phase I trial of 4'-O-tetrahydropyranyl-doxorubicin (THP)--a multi-institutional cooperative study].

A phase I trial of a new anthracycline derivative, 4'-O-tetrahydropyranyldoxorubicin (THP), was conducted in 54 patients with various advanced solid tumors and malignant lymphomas. Starting dose was 5 mg/m2 i.v. and dose escalations were made by a modified Fibonacci search scheme. There were 40 evaluable courses. The dose-limiting toxic effect was leukopenia which was dose-related and reversible. The maximum tolerated dose for a single i.v. injection was estimated to be 55 mg/m2. The median nadir day for leukopenia was day 12 with recovery occurring 13 days (median) after reaching the nadir. Thrombocytopenia was less commonly observed than leukopenia. Other toxic effects were mild gastrointestinal disturbances, fever and general malaise. Ventricular extrasystole was observed in a case of pancreatic cancer who received 5 mg/m2 of the drug. There were no cases with alopecia, or with hepatic or renal dysfunction. With regard to objective tumor response, CR was observed in 2 cases with NHL, and MR in 2 cases with lung cancer, and 1 case each with breast cancer and NHL. Response occurred at a dose of more than 35 mg/m2. The recommended dose schedule for phase II trial is 35-45 mg/m2 by single i.v. injection at 3-4-week intervals.

[Transition of occurrence of anisakiasis and its paratenic host fishes in Japan, with pathogenesis of anisakiasis].

The number of anisakiasis according to a nation-wide statistical survey totaled 4682 cases until June 1987 in Japan. Of these 4296 cases are gastric anisakiasis (including 215 cases of gastric terranovasis), 375 intestinal anisakiasis and 11 extra-gastrointestinal anisakiasis. Pseudoterranova larva does not invade the intestine, and worms are vomited in most cases. Clinical diagnosis of intestinal anisakiasis is more difficult than that of gastric anisakiasis, and it also is hard to find the worm itself in histopathological examination. Therefore the number of actual intestinal anisakiasis is probably 3 times more than that of the reported cases. The cases of gastrointestinal-wall perforation by worms are increasing, which means immune response by granuloma formation is important. The catches of paratenic hosts and the rate of infection vary with year. In addition, the kind of paratenic host fishes are different between the southern and northern areas of Japan. The paratenic hosts reported by patient are closely related to the catches of kinds of those fishes in the respective areas. Recently, cases by eating sardine are increasing in the southern area. Urticaria as a complication is related to the diagnostic rate, and intraperitoneal bacterial infection by the gastrointestinal perforation by worms is closely related to the prognosis.

Possible involvement of peripheral-type benzodiazepine receptors in erythroid differentiation of human leukemia cell line, K562.

Possible involvement of the peripheral-type benzodiazepine receptor (PBR) in hemin/protoporphyrin-induced erythroid differentiation of human leukemia K562 cells was investigated by the use of the ligands, diazepam and PK11195. Diazepam itself exhibited differentiation-inducing activity on K562 cells. The PBR-specific antagonist, PK11195, dose-dependently inhibited both diazepam-induced and hemin/protoporphyrin-induced K562 cell differentiation. The results imply that PBR is involved in the erythroid differentiation of K562 cells.

Erythroid differentiation-inducing activity of protoporphyrin IX and its analogs on human leukemia K562 cell line.

Protoporphyrin IX, which had been regarded as ineffective in cell differentiation, induced erythroid differentiation of K562 cell line, and its analogs such as hematoporphyrin IX, mesoporphyrin IX, deuteroporphyrin IX and protoporphyrin IX dimethyl ester were also differentiation inducers. The differentiation induced by these compounds was augmented by a retinoid, as is hemin-induced differentiation.

Hemin-induced erythroid differentiation of human myeloleukemia K562 cell line and its modification by bioresponse modifiers.

We have found that protoporphyrin IX, which had been regarded as inactive, induces erythroid differentiation. The differentiation-inducing activities of various hemin-related compounds, including hematoporphyrin IX, mesoporphyrin IX, deuteroporphyrin IX and protoporphyrin IX dimethyl ester, suggested certain structural requirements for the activity: 1) the iron moiety of hemin is not essential, and 2) the propionic acid side chains of hemin play an important role. In addition, we have examined the influence of some bioactive factors on hemin/protoporphyrin IX-induced differentiation of K562 cell line. Retinoids and tubulin-disruptors dose-dependently enhanced hemin/protoporphyrin IX-induced differentiation of K562 cells, though they did not themselves induce differentiation. Retinoid antagonists suppressed hemin-induced differentiation. The effects of hemin and/or retinoids on the mRNA expressions of oncogenes (c-myc and c-myb) and retinoic acid receptor genes (rar alpha and rar beta) of K562 cells were analyzed. We also examined the possible involvement of peripheral-type benzodiazepine receptor (PBR) in hemin/protoporphyrin IX-induced differentiation of K562 cells by the use of its ligands. Diazepam itself was revealed to possess differentiation-inducing activity on K562 cells. The PBR-specific ligands modified hemin-induced differentiation. These results suggest a requirement for retinoids (or retinoid-like cofactors) for hemin/protoporphyrin IX-induced differentiation of K562 cells and the involvement of PBR in erythroid differentiation of K562 cell line.