Enhancement of activated beta1-integrin expression by prostaglandin E2 via EP receptors in isolated human coronary arterial endothelial cells: implication for the treatment of Kawasaki disease.

OBJECTIVE: Plasma prostaglandin E(2) (PGE(2)) levels are markedly elevated in acute Kawasaki disease (KD). We evaluated the function of the EP receptors in the expression of activated beta(1)-integrin stimulated by PGE(2) in human coronary arterial endothelial cells (HCAEC). METHODS: We determined the mRNA expression of the PGE(2) receptors, EP receptors (EP(1-4)) in HCAEC by RT-PCR and protein expression by Western blotting. We evaluated the function of the EP receptors in the expression of activated beta(1)-integrin stimulated by PGE(2) in HCAEC, using antagonists and agonists of the EP receptors, by flow cytometry. RESULTS: RT-PCR revealed mRNAs for all four EP receptors in HCAEC. Western blotting demonstrated EP(1), EP(2) and EP(3) expression in HCAEC. The EP(2) and EP(3) agonists enhanced the expression of activated beta(1)-integrin in HCAEC. The potency of the EP(2) agonist was significantly greater than that of the EP(3) agonist. Pretreatment with the EP(1), EP(2) and EP(3) antagonists inhibited the expression of activated beta(1)-integrin induced by PGE(2) in HCAEC. The potency of the EP(2) antagonist was significantly greater than that of the EP(1) and EP(3) antagonists. CONCLUSIONS: Our results suggest that PGE(2) mainly induces the activation of beta(1)-integrins via the EP(2) receptor in HCAEC. Our results further suggest that the EP(2) antagonist modulates the inflammatory response during KD vasculitis.

[Long-term results of arterial switch operation].

Surgical results and late outcome in 202 patients who had undergone arterial switch operation from 1984 to 1997 were investigated. Actuarial survival was 90.6% at 10 years and 90.0% at 20 years. Fifty-two patients (25.7%) underwent reoperation for pulmonary stenosis and 7 patients (3.5%) had aortic valve replacement. Freedom from re-intervention was 71.9% at 10 years and 60.4% at 20 years. Using xeno-pericardial patch for pulmonary reconstruction was strong predictor for postoperative pulmonary stenosis. Coronary ischemic event was rare but some patients showed electorocardiogram (ECG) change on exercise and hypoplastic left coronary artery. Cardiopulmonary function was almost normal in long term survivors.

Cytokine-related and sodium channel polymorphism as candidate predisposing factors for childhood encephalopathy FIRES/AERRPS.

Febrile infection-related epilepsy syndrome (FIRES), or acute encephalitis with refractory, repetitive partial seizures (AERRPS), is an epileptic encephalopathy beginning with fever-mediated seizures. The etiology remains unclear. To elucidate the genetic background of FIRES/AERRPS (hereafter FIRES), we recruited 19 Japanese patients, genotyped polymorphisms of the IL1B, IL6, IL10, TNFA, IL1RN, SCN1A and SCN2A genes, and compared their frequency between the patients and controls. For IL1RN, the frequency of a variable number of tandem repeat (VNTR) allele, RN2, was significantly higher in the patients than in controls (p=0.0067), and A allele at rs4251981 in 5' upstream of IL1RN with borderline significance (p=0.015). Haplotype containing RN2 was associated with an increased risk of FIRES (OR 3.88, 95%CI 1.40-10.8, p=0.0057). For SCN1A, no polymorphisms showed a significant association, whereas a missense mutation, R1575C, was found in two patients. For SCN2A, the minor allele frequency of G allele at rs1864885 was higher in patients with borderline significance (p=0.011). We demonstrated the association of IL1RN haplotype containing RN2 with FIRES, and showed a possible association of IL1RN rs4251981 G>A and SCN2A rs1864885 A>G, in Japanese patients. These preliminary findings suggest the involvement of multiple genetic factors in FIRES, which needs to be confirmed by future studies in a larger number of FIRES cases.

Alteration of activities of delta-aminolevulinic acid synthase, delta-aminolevulinic acid dehydratase and delta-aminolevulinic acid dehydratase inhibitor in the bone marrow cells of lead poisoned rats.

A new modified method for the determination of delta-aminolevulinic acid (ALA) dehydratase activity has been developed. This modified method involves addition of 0.1 mM Zn2+ and then preheating the enzyme solution at 60 degrees C for 5 min before following commonly used procedures. The ALA dehydratase activity in the peripheral blood of lead poisoned rats, determined by this procedure, was approximately 7.5-fold higher than control values. The activity of ALA dehydratase in the bone marrow cells of lead poisoned rats increased approximately 2-fold. These increases were accompanied by a simultaneous decrease in an inhibitor specific for ALA dehydratase. Consequently, these results suggest that ALA dehydratase is induced after the reduction of ALA dehydratase activity, and that ALA dehydratase activity is controlled by a new specific ALA dehydratase inhibitor.

Effect of protein nutrition throughout gestation and lactation on growth, morbidity and life span of rat progeny.

Thirteen Fischer strain rats were divided randomly into 3 groups soon after became pregnant and fed a 10%, 18%, or 36% casein diet throughout pregnancy and lactation. At weaning, female pups from the dams were divided into 3 further groups which were fed throughout life 10%, 18%, or 36% casein diet. The animals were weighed regularly and given a full autopsy after death. DNA was determined in the cerebrum and cerebellum of progeny at 7, 15 and 50 weeks of age respectively. The level of protein intake of the dams did not affect the litter-size, but did affect the body weight of pups at weaning. When fed a high protein diet after weaning, pups from dams fed a low protein diet weighed less throughout life than those from dams fed a high protein diet. A similar effect of protein intake was observed on the tail length. Pups of dams fed a low protein diet ate a larger amount of diet per unit body weight after weaning than those of dams fed at a high level of protein. Pups at 7, 15, and 50 weeks of age respectively from dams fed 10% casein diet had a lighter cerebrum and cerebellum than pups of the same ages from dams fed respectively 18% and 36% casein diet. They also had less total DNA in the cerebrum and cerebellum than the latter. At the ages of 1 and 2 years respectively, the survival rates of rats fed the same diet as their mothers tended to be greater than those of rats fed diet different from that of their mothers. They also tended to live longer than the latter: 4 of 60 rats lived for over 1,000 days, and 3 of these were fed the same diet as that of their mothers. Lesions observed at death did not differ in different groups. However, the incidence of lesions of the kidney and the number of tumors seemed to be greater in groups fed a high protein diet.

Protein nutrition and aging.

Twenty-four female rats consisting of 6 sets of litters were used for the experiment. After weaning, rats were divided into 4 groups and fed with 10, 18, 27 and 36% casein diet. Effect of protein nutrition on aging was examined from the anthropometric and biochemical viewpoint. The difference of growth in body weight, and that of the urinary excretion of creatinine and 17-ketosteroids observed in the early period of growing disappeared by the end of growth period. From 1 year after birth, serum alkaline phosphatase activity and serum cholesterol were also measured. However, effect of protein nutrition on these parameters was not clear due to the scatter of data. The difference in diet did not affect life span in the present experiments, but the effect of the variance of litters on it seemed to be significant between the rats fed 18% casein diet and those fed 10% casein diet. Rats fed high-protein diet had a great number of lesions in the kidney and hypophysis, and often an incidence of tumors.

[The effect of magnesium on calcium oxalate crystal shapes generated with glycosaminoglycan].

Our previous report has shown that the shapes of calcium oxalate crystals formed in a supersaturated solution with glycosaminoglycan varied with glycosaminoglycan species. The present study was conducted to demonstrate the effect of magnesium on the shapes of calcium oxalate crystals with glycosaminoglycan by optical and scanning electron microscopy. In the presence of magnesium, the phenomenon of the plate- or sheet-like crystals parallel growing on the surface of other crystals was observed with each glycosaminoglycan and was enhanced with hyaluronic acid, chondroitin sulfate, heparan sulfate or heparin, more marked in this order. On the other hand, with heparan sulfate or heparin the stratification of the plate-like crystals was observed, whereas with hyaluronic acid or chondroitin sulfate it was not observed. Moreover, the stratification of the crystals with heparin was stimulated more markedly with increasing magnesium concentrations, and the lamella-shaped crystals at low heparin concentrations in the presence of magnesium was similar to the crystals at high heparin concentrations. These results suggest that magnesium stimulates the effect of glycosaminoglycan on calcium oxalate crystal shapes.

Cysteinyl leukotrienes enhance tumour necrosis factor-alpha-induced matrix metalloproteinase-9 in human monocytes/macrophages.

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is an important enzyme responsible for airway remodelling. Monocytes/macrophages have a cysteinyl leukotriene 1 (cysLT1) receptor, but its function is poorly understood. OBJECTIVE: To elucidate the function of the cysLT1 receptor of human monocytes/macrophages in MMP-9 production. METHODS: We examined the effect of cysLTs (LTC4, -D4 and -E4) on TNF-alpha-induced MMP-9 production in THP-1 cells, a human monocytic leukaemia cell line and peripheral blood CD14+ monocytes/macrophages. In addition, we examined the effect of pranlukast, a cysLT1 receptor antagonist, on the enhancement of TNF-alpha-induced MMP-9 production by cysLTs. RESULTS: ELISA revealed that LTC4 and -D4, but not -E4, enhanced TNF-alpha-induced MMP-9 production in THP-1 cells and peripheral blood CD14+ monocytes/macrophages. Real-time polymerase chain reaction demonstrated that LTC4 and -D4, but not -E4, increased MMP-9 mRNA expression induced by TNF-alpha in THP-1 cells. Moreover, we demonstrated that pranlukast completely inhibited the enhancement of TNF-alpha-induced MMP-9 production by LTC4 and -D4 in THP-1 cells and peripheral blood CD14+ monocytes/macrophages. CONCLUSION: LTC4 and -D4 enhanced the TNF-alpha-induced MMP-9 production via binding the cysLT1 receptor in human monocytes/macrophages. Pranlukast inhibited the enhancements by LTC4 and D4.

Effects of excess intake of leucine and valine deficiency on tryptophan and niacin metabolites in humans.

Nineteen healthy humans (2 men and 17 women) served as experimental subjects in 4 experiments using diets having different levels of leucine and also a valine-deficient diet. The effect of an excess intake of leucine, with and without addition of vitamin B-6, and the effect of a deficiency of valine on urinary excretions of N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, nicotinic acid, quinolinic acid, and 5-hydroxyindole acetic acid, and on the level of plasma amino acids were investigated. There was no effect of leucine on the excretion of these metabolites, but a marked decrease in the plasma (or serum) valine level was observed. The same decrease was seen when a valine-deficient diet was fed.

Increase of delta-aminolevulinic acid dehydratase (ALAD) in rat erythrocytes in lead poisoning.

The well known fact that the activity of delta-aminolevulinic acid dehydratase (ALAD: EC 4.2.1.24) is reduced in red cells of animals with lead poisoning was found to be upset, by using a modified method of Gibson's original procedure, for determination of activated ALAD activity. The modified method involves addition of 0.2 mM Zn2+ and then preheating the enzyme solution at 60 degrees C for 5 min before following Gibson's original procedure. With this methodological modification, the ALAD activity of erythrocytes of rats poisoned with lead was found increased. Furthermore, the enzyme was purified from the peripheral blood of lead-poisoned rats. ALAD protein in peripheral blood was also determined by single radial immuno diffusion using rabbit anti-serum raised against rat liver ALAD. As the result, the ALAD activity obtained from the modified method was found to be directly proportional to the absolute amount of enzyme proteins determined both by chemically and immunochemically. The modified method for measuring true ALAD content in blood cells in lead poisoning is more reliable than previous ones.

[An operative case of spermatic cord rhabdomyosarcoma--with a review of similar cases in Japan].

A 17-year-old Japanese boy was admitted with the complaint of painless, progressive enlargement of the left scrotal contents during the preceding two months. The tumor was firm and nontender without transillumination. Orchitectomy with high ligatation of the spermatic cord was performed. The tumor, weighing 13.1 g, was gray, solid, located at the distal part of the spermatic cord, and separated from the testis and epididymis. The pathological diagnosis showed left spermatic cord rhabdomyosarcoma of the combined embryonal and alveolar type. All of the retroperitoneal lymph nodes dissected were negative for metastasis. The patient was treated with actinomycin D and vincristin postoperatively, and is well three years after the operation. Ninety-two cases of intrascrotal rhabdomyosarcoma reported in Japan are reviewed.