Safety and feasibility of intra-arterial nicardipine for the treatment of subarachnoid hemorrhage-associated vasospasm: initial clinical experience with high-dose infusions.

BACKGROUND AND PURPOSE: Delayed cerebral ischemia from vasospasm is a major complication after aneurysmal subarachnoid hemorrhage (SAH), but complications and/or low efficacy are associated with current therapy. We report our initial experience with intra-arterial use of a calcium channel blocker, nicardipine. MATERIALS AND METHODS: A retrospective review of a consecutive series of patients with clinical and angiographic vasospasm treated with intra-arterial nicardipine was performed. Standard criteria for definition of significant, intractable vasospasm after aneurysmal SAH were used. After catheter angiographic confirmation of vasospasm, arteries showing severe narrowing were targeted for superselective catheterization. Nicardipine was infused at a high dose rate (0.415-0.81 mg/min). Contrast injections were performed at 2-5-mg intervals to assess effective response (a 60% increase in arterial diameter of the most severely decreased in caliber vessel compared with the very first angiographic run). RESULTS: Eleven consecutive patients underwent a total of 20 procedures; most had SAH with high Hunt and Hess grades (III or IV). All had depressed level of consciousness; others had paresis (7/20, 35%), aphasia (1/20, 5%), and facial nerve palsy (1/20, 5%). Between 10 and 40 mg of nicardipine was used. A 60% increase in diameter of the main affected artery compared with the initial diameter measured in the initial angiographic run was achieved in all procedures. Clinical improvement (resolved focal symptoms or increased Glasgow Coma Score) occurred in 10 of 11 patients (91%). One patient died from complications of the initial hemorrhage. No complications occurred after 16 of 20 procedures (80%); minor complications without sequelae occurred after the remaining procedures. Follow-up of at least 2 months in 10 survivors revealed minor or no deficits in most patients with a Glasgow Outcome Score of 1 or 2 in 9 of 10 patients (90%). CONCLUSION: In this small series, high-dose intra-arterial nicardipine infusion to treat SAH-associated vasospasm seems to be safe and effective.

Application of sucrose-gradient centrifugation for selective isolation of Nocardia spp. from soil.

AIMS: To devise and evaluate a method for selective isolation of the less abundant actinomycetes, Nocardia spp. in soil. METHODS AND RESULTS: This newly developed method is based on differentiating Nocardia from other actinomycete taxa by centrifugation. A water suspension of air-dried soil is centrifuged through a gradient consisting of 10, 20, 30, 40 and 50% sucrose at 240 x g for 30 min. The 20% sucrose layer, which is enriched with Nocardia spp., is then diluted and plated on humic acid-vitamin agar supplemented with antibacterial agents. The proposed method consistently achieved selective isolation of Nocardia spp. in all 14 soil samples tested, which accounted for 5-89% of the total microbial population recovered. Tentative taxonomic characterization based on a restriction fragment length polymorphism (RFLP) analysis of the 16S ribosomal DNA suggested that many of the soil isolates could belong to N. asteroides, N. salmonicida or N. uniformis. CONCLUSIONS: Differential centrifugation can successfully and efficiently isolate soil Nocardia populations that are suppressed by conventional dilution plating approaches. SIGNIFICANCE AND IMPACT OF THE STUDY: The development and application of new methodologies with which to isolate less-explored actinomycete taxa is important for improving our knowledge about their taxonomy, ecology and industrial applications.

An integrated method for the enrichment and selective isolation of Actinokineospora spp. in soil and plant litter.

AIMS: To devise and evaluate a method for isolating the rare, zoosporic actinomycetes, Actinokineospora spp. in soil and plant litter. METHODS AND RESULTS: The newly developed method consists of two enrichment stages followed by plating on a selective medium. The source material is initially incubated with calcium carbonate to multiply the population of Actinokineospora spp., and is then air-dried. The second stage consists of rehydration-centrifugation, in which the amended substrate is immersed in phosphate buffer-soil extract to liberate actinomycete zoospores, and nonmotile microbial associates are then eliminated by centrifugation. Portions of the supernatant enriched with zoospores are plated on humic-acid vitamin agar supplemented with fradiomycin, kanamycin, nalidixic acid and trimethoprim. We examined 39 soil and plant-litter samples taken from fields, forests and stream banks. The proposed method consistently enriched and selectively isolated Actinokineospora spp. in 17 samples. Evidence for antimicrobial activity was found in most of the isolates. CONCLUSION: A combination of enrichment and a medium containing selective antibiotics can be used successfully for efficient isolation of certain rare actinomycete taxa. SIGNIFICANCE AND IMPACT OF THE STUDY: The development of new methodologies with which to isolate rare actinomycetes is of great importance to extend our understanding of their ecology, taxonomy and bioactivity.

Plasma and erythrocyte levels of trace elements and related antioxidant enzyme activities in low-birthweight infants during the early postnatal period.

UNLABELLED: To discover the relationship between antioxidant enzyme activities and trace elements in low-birthweight infants during the early postnatal period, we analysed catalase (CAT), CuZn-superoxide dismutase (CuZn-SOD) and glutathione peroxidase (GSH-Px) activities in erythrocytes. and compared them with Fe, Cu. Zn and Se levels in plasma, and Cu, Zn and Se levels in erythrocytes until 16 wk after birth. Thirteen low-birthweight infants whose mean birthweight and gestation were 1520 +/- 293 g and 32.0 +/- 2.8 wk were enrolled in this study. All infants were without chronic complications, well nourished, and predominantly fed standard formula or preterm formula based on cow's milk, commercially available in Japan. Cu and Zn levels in erythrocytes did not decline after birth, in contrast to a temporal decrease in plasma Zn. Erythrocyte CAT activity was significantly higher at 16 wk than that at birth or 4 wk of age. Erythrocyte CuZn-SOD activity did not change throughout the study period. Only Se in plasma and erythrocytes decreased remarkably after birth, which resulted in a significant decline in erythrocyte GSH-Px activity at 8 and 16 wk (11.2 +/- 2.0 and 11.0 +/- 1.1 U/g Hb) compared to that at birth (12.4 +/- 2.1 U/g Hb). CONCLUSION: During the early postnatal period. erythrocyte CAT and CuZn-SOD activities did not decrease in formula-fed, low-birthweight infants. Japanese commercial formula not supplemented with Se, however, caused postnatal plasma and erythrocyte Se deficiency and a concomitant decline in erythrocyte GSH-Px activity in the same subjects.

Ascorbic acid improves postischemic vasodilatation impaired by infusion of soybean oil into canine iliac artery.

This study was conducted to (a) assess postischemic vasodilatation by changes in the vascular cross-sectional area using simultaneous intravascular two-dimensional and Doppler ultrasound before and after the infusion of Intralipid (Pharmacia & Upjohn, Peapack, NJ, U.S.A.); (b) evaluate how antioxidant ascorbic acid modifies the effects of Intralipid on postischemic vasodilatation: and (c) clarify the changes in plasma nitrite and nitrate (NOx-) levels after the infusion of Intralipid with and without ascorbic acid. Twenty-eight mongrel dogs were used to measure for vascular cross-sectional area and average instantaneous peak velocity in the iliac arteries after the 5-min occlusion of the arteries. Postischemic vasodilatation was impaired after the infusion of Intralipid (20%, 2 ml/kg) and this impaired response was reversed by the co-administration of ascorbic acid (30 mg/kg). NG-monomethyl-L-arginine completely abolished postischemic vasodilatation. Plasma NOx levels were significantly reduced after the infusion of Intralipid compared with baseline (11.6+/-0.4 vs. 12.9+/-0.3 microM, p = 0.025) and after infusion of Intralipid with ascorbic acid compared with baseline (11.8+/-0.5 vs. 13.1+/-0.4 microM, p = 0.047). We concluded that ascorbic acid reverses the endothelial dysfunction induced by Intralipid without increasing plasma NOx- levels and that deactivation of nitric oxide by oxidative stress is a primary contributor to Intralipid-induced impaired vasodilation.

Application of a method incorporating differential centrifugation for selective isolation of motile actinomycetes in soil and plant litter.

The present paper describes a simple enrichment technique which enables rapid and selective isolation of diverse zoosporic actinomycete genera directly from soil and plant litter. This technique, designated the rehydration and centrifugation (RC) method, consists of immersing the air-dried source material in 10 mM phosphate buffer containing 10% soil extract, letting the preparation stand at 30 degrees C for 90 min, followed by centrifugation of the fluid at 1,500 x g for 20 min. Portions of the supernatant containing actinomycete zoospores are plated on the humic acid-vitamin agar which is supplemented with nalidixic acid and trimethoprim as the selective inhibitors for Gram-negative bacteria and bacilli. The phosphate buffer-soil extract solution significantly promoted liberation of motile zoospores from the source material. The centrifugation stage greatly eliminated streptomycetes and other non-motile actinomycetes from the liquid phase, thereby facilitating selective growth of rare, motile actinomycetes on the isolation plates subsequent to inoculation. Ten different soil and leaf-litter samples, taken from fields, forests, and stream banks, were examined. The RC method consistently achieved preferential isolation of motile actinomycetes in all samples, which accounted for 37-86% of the total microbial population recovered. The most frequently isolated motile actinomycetes were Actinoplanes and Dactylosporangium. Strains of Actinokineospora, Catenuloplanes and Kineosporia were also recovered, depending on the nature of the samples examined. Other motile actinomycetes that were occasionally isolated in small numbers included Actinosynnema, Geodermatophilus and Sporichthya.

Effects of ubiquinone enriched diet on deterioration of mitochondrial respiratory function caused by fried beef derived mutagenic factor in rats.

2-Amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP), a potent mutagenic agent produced during thermal processing of meats, shows detrimental effects on mitochondrial respiratory function. Effects of dietary ubiquinone-10 was examined on PhIP-induced mitochondrial respiratory dysfunction in rats. Rats fed with 0.2% ubiquinone-10 diet mitigated the PhIP-induced decrease in mitochondrial complex I activities observed in rat heart, diaphragm, and psoai major. Therefore, dietary anti-oxidants such as ubiquinone-10 could cancel out the unfavorable effects of PhIP and contribute to the preservation of mitochondrial function.

[Two cases of acute disseminated encephalomyelitis with lesions in the thalamus or basal ganglia on MRI].

Lesions in the thalamus or basal ganglia have rarely been reported in acute disseminated encephalomyelitis (ADEM). We experienced 2 cases of ADEM, in which MRI showed lesions in the thalamus or basal ganglia. Case 1, a 4-year-old boy, had gait disturbance, hyperesthesia and hyperreflexia. MRI (T2 weighted image) showed multiple high intensity areas in the right frontal lobe, bilateral parietal lobes and bilateral thalami. Case 2, a 4-year-old girl, complained of gait disturbance following a febrile episode, and displayed hyperreflexia. Several days later, she had visual disturbance of the left eye. MRI (T2 weighted image) revealed multiple high intensity areas in the dentate nucleus of left cerebellum, left occipital lobe, bilateral caudate nuclei, and the anterior part of bilateral lenticular nuclei. In both cases, CT could not demonstrate these lesions. Both of them were treated with corticosteroid and recovered rapidly. They had no recurrence. MRI is useful in diagnosis and follow-up of ADEM and may reveal lesions other than cerebral or cerebellar white matters.

Growth inhibition of human fibroblasts by epidermal growth factor in the presence of arachidonic acid.

The effect of epidermal growth factor on growth of human fibroblasts was investigated in serum-free medium supplemented with various fatty acids. When linoleic acid, arachidonic acid, or eicosapentaenoic acid was added, each inhibited epidermal growth factor-induced cell growth and showed cytotoxicity at high concentrations (greater than 10 microM). This cytotoxic effect was not observed in the presence of indomethacin, suggesting that prostaglandin production is important in mediation of the growth inhibition. Prostaglandin E2 was increased more than ten thousand times by epidermal growth factor in combination with arachidonic acid.

Treatment of chronic liver injury in mice by oral administration of xiao-chai-hu-tang.

Oral administration of Xiao-Chai-Hu-Tang, the extract of a mixture of seven herbs, attenuated the hepatic fibrosis developed in mice after repeated doses of carbon tetrachloride. Its pre-administration reduced the derangement of liver function tests seen after a single dose of carbon tetrachloride as well as that seen after d-galactosamine intoxication. Xiao-Chai-Hu-Tang may be effective in the treatment of chronic liver injury through hepatocytoprotection.

Adverse effects of anti-tumor drug, cisplatin, on rat kidney mitochondria: disturbances in glutathione peroxidase activity.

This study was designed to clarify mechanisms responsible for cisplatin-induced nephrotoxicity together with the effect of selenium. Rats were divided into 3 groups: the cisplatin group; cisplatin (60 mg/kg) was administered intraperitoneally once, the cisplatin + Se group; cisplatin (60 mg/kg) once, and selenious acid (10 mumol/kg) were administered intraperitoneally once a day for 5 consecutive days, the control group; untreated. In each group, mitochondrial respiratory function, enzymic activities in mitochondrial respiratory chain and glutathione peroxidase, and plasma creatinine and BUN contents were measured. In the cisplatin group, decreases in mitochondrial respiratory function, enzymic activities in the respiratory chain and glutathione peroxidase, and increases in plasma creatinine and BUN contents were observed compared with the control group, while the cisplatin + Se group lessened these impairments. These results suggested that cisplatin-induced nephrotoxicity was closely related to mitochondrial dysfunction through the impairment of glutathione peroxidase. This toxicity might be ascribed to free radical mediated-injury. We propose here that, with selenium, higher dose administration of cisplatin to patients might be applicable.

Cholesterol metabolism in inherently scorbutic rats (ODS rats).

We observed the cholesterol metabolism of a colony of Wistar rats with a hereditary defect in vitamin C synthesizing ability (the ODS (osteogenic disorder-Shionogi) rats) in six kinds of experiments. Female ODS rats aged 36 days had a low HDL (high-density lipoprotein)-cholesterol level in serum as compared with age-matched control rats in spite of the absence of scorbutic symptoms. Female ODS rats aged 63 days which revealed severe scorbutic symptoms had a very low HDL-cholesterol level (mean value; 17 mg/dl). And male ODS rats, whose lives had been prolonged by supplementing with L-ascorbic acid, also had lower serum HDL-cholesterol and had increased total cholesterol in serum and liver when the acid supplement dose was relatively insufficient. On the other hand, we examined HDL2- and HDL3-cholesterol levels in serum to determine the mechanism of low HDL-cholesterol. As a result, we observed a low HDL2-cholesterol level in ODS rats but normal HDL3-cholesterol level. But the authors observed no decrease of LCAT (lecithin: cholesterol acyltransferase) activity in serum of ODS rats. These results could be due to disturbance of lipid metabolism in a vitamin C-deficient condition, that is to say, there might be abnormalities of the cholesterol excretion pathway of bile acid from liver, and maturity of the HDL-cholesterol particle due to other factors except that of LCAT activity.

Risk factors of atherogenicity in inherently scorbutic rats (OD rats).

We observed the pathophysiological state of a colony of Wistar rats with a hereditary defect in vitamin C-synthesizing ability (the OD rat) in three kinds of experiments. In experiment 1, OD rats that were not supplemented with L-ascorbic acid (ASA) were killed 36 days after birth. In experiment 2, OD rats that were not supplemented with AsA were killed 63 days after birth. In experiment 3, OD rats that were supplemented with sufficient ASA (100 mg/100 g body weight/day) for 16 days from 47 days after birth were assayed 63 days after birth. From analyses of serum chemistry, platelet aggregability and counts of blood cells, some atherogenic risk factors were elucidated, that is, there was a decrease of high density lipoprotein in serum and an increase of platelet aggregability. Furthermore, it was observed that OD rats have some characteristics different from latent vitamin C-deficient animals from the viewpoint of malnutrition.

What are the contents of a hydrocele testis?

Relatively large, uninfected hydroceles of the testis were investigated as to the composition of their fluid. The fluid was different from serum and urine, having the urea nitrogen, uric acid, Na and K levels comparable to but the total protein and inorganic phosphorus levels higher than in normal serum. The analysis of hydrocele contents may be useful in ruling out the possibility of another retention tumor, but not useful in determining whether a hydrocele is communicating with the peritoneal cavity.