Activation of alveolar phospholipase A2 after hydrochloric acid aspiration in rats.

PURPOSE: The present study was carried out to determine phospholipase A2 (PLA2) activity in the bronchoalveolar lavage fluid (BALF) in rats subjected to HCI aspiration. MATERIALS AND METHODS: Rats were allocated into one of five groups. Groups H-1 and H-3 received instillation of HCI into lungs. Groups S-1 and S-3 received saline instead of HCI. Group C received no instillation. BAL was performed according to the protocol, that is, 1 hour after the instillation in groups H-1 and S-1, 3 hours after the instillation in groups H-3 and S-3, and arbitrarily in group C. Obtain BALF was analyzed for the protein concentration, PLA2 activity, and the molecular mass of PLA2. RESULTS: The protein concentration in BALF showed an increase in groups H-1 and H-3. PLA2 activity decreased in group H-1, but increased in group H-3, compared with groups S-1 and S-3, respectively. PLA2 in groups C and H-1 revealed a high molecular mass (HM), but that in group H-3 revealed a low molecular mass (LM). CONCLUSIONS: There is an increase in the alveolar LM-PLA2 at inflammatory phase after HCI aspiration, suggesting the pathophysiologic role of LM-PLA2 in the acute lung injury.

Cerebral arterial spasm. II. Etiology and treatment of experimental cerebral vasospasm.

Delayed cerebral vasospams is caused by excessive accumulation of dopamine-beta-hydroxylase (DBH) and noradrenaline in cerebral vessel walls. This study demonstrates the mechanisms of delayed spasm, particularly the role of red blood cell components, and the successful relief of delayed cerebral vasospasm. Spasmogenic substances which contained a heme component, such as methemoglobin, methemalbumin and catalase enhanced DBH activity in human serum as measured by a one step chemical spectrophotometric assay. The concentration which gave the highest DBH activity caused the maximum constriction of the basilar artery, when the substances were applied topically. Among components of red cells, methemoglobin, methemalbumin, catalase and nicotinamid adenin dinucleotide (NADH) caused constriction of basilar artery in cats, when applied topically, whereas hematin, hemin and bilirubin caused no significant spasm. An oxyhemoglobin solution obtained by mixture with methemoglobin and ascorbic acid produced no significant vascular spasm either. Relief of delayed cerebral vasospasm was obtained with topical application of specific alpha adrenergic blocking drug such as phenoxybenzamine, specific inhibitors of DBH such as fusaric acid, o-phenanthroline and alphaalpha' dipyridyl beta2 adrenergic stimulants such as salbutamol, and a phosphodiesterase inhibitor, ascorbic acid.

Cerebral aterial spasm. I. Adrenergic mechanism in experimental cerebral vasospasm.

This study demonstrates that an adrenergic mechanism plays an important role in producing the delayed cerebral vasospasm which follows subarachnoid hemorrhage. Results were as follows: 1. Experimental subarachnoid hemorrhage (SAH) was produced by injection of fresh arterial blood into the cisterna magna in cats. The cerebral vasospasm was shown angiographically to be biphasic in nature: immediate constriction lasting 1 h and marked prolonged spasm occurring between the 3rd and 5th day after SAH. The amount of noradrenaline (NA) and dopamine-beta-hydroxylase (DBH) activity decreased over a period of 24 h both within the wall of the basilar artery and in the locus ceruleus and then gradually increased, reaching a maximum on the 3rd day after SAH. 2. Topical application of spasmogenic substances (NA and blood) produced a marked constriction of the hypersensitive basilar artery on the 3rd day after SAH. 3. 6-Hydroxydopamine (6-OHDA) injection into the cisterna magna produced prolonged vasocilatation. The dilated vessel responded with mild transient constriction after the topical application of NA or fresh blood. DBH activity and NA concentration in the vessels, locus ceruleus and medial hypothalamus decreased markedly on the 3rd day after the cisternal injection of 6-OHDA. 4. Various spasmogenic substances (i.e. serotonin, NA, prostaglandins and methemoglobin) were measured in a mixture of equal volume of CSF and blood in cats. ONly the serotonin in the mixed fluid produced vasoconstriction. Spasmogenic substances decreased markedly in the mixed fluid incubated for 3 days at 37 degrees C, and none of these substances apart from methemoglobin was present in a concentration sufficient to produce constriction of vessels. 5. These results suggest that early spasm is induced by serotonin around the arteries of the cranial base, and delayed spasm might be caused by hyperreaction of cerebral vessels to spasmogenic substances such as methemoglobin, during the accumulation of excess NA in the cerebral vessel wall.

Methemoglobinemia induced by chlorphenamidine.

A 76-year old farmer ingested 100 g of chlorphenamidine (Galectron), a plant acaricle, for the purpose of suicide. Gastric lavage was performed and the patient survived. Methemoglobinemia was noted after emergency treatment and was still present at 20 hours after ingestion of the compound. The patient was lethargic for at least 50 hours. Moderate neutrophilic leukocytosis and kidney injury were observed.

[Intercostal nerve cross-anastomosis and DREZ-tomy in a patient with brachial plexus avulsion].

An 18-year-old male with a right brachial plexus injury caused by a motorcycle accident was admitted on October 13, 1988. A detailed examination revealed that the brachial plexus was totally injured. The axon reflex test suggested that the lesion sites were postganglionic in the C5 and C6 nerves, and preganglionic in the C7, C8 and Th1. On December 14, 1988, intercostal nerve cross-anastomosis was performed in the hope that a lost motor function of the right upper extremity could be restored as a first step. An electromyogram 6 months after this anastomotic operation demonstrated synkinesis between the biceps brachialis and the intercostal muscles during deep inspiration. Fifteen months after this operation, active voluntary muscle discharges which were higher than before in amplitude, were provoked. The right elbow flexion was gradually restored independently of respiration. On the other hand, intractable pain with a persistent severe tingling sensation appeared approximately one week after the injury on the lesioned upper extremity. The pain rapidly increased in severity. The DREZ-tomy from C5 to Th1 was performed on March 7, 1989. This has brought complete relief of the intractable pain. A brief discussion was given concerning some aspects of the brachial plexus avulsion and DREZ-tomy.

[Long-term observation of individual erythrocyte sedimentation rate and its relation to other tests variation].

We have studied individual changes of erythrocyte sedimentation rate (ESR) in 30 so-called healthy subjects by examining them 2-4 times a year for 14-20 years. The results showed big differences in the same subjects during these period, and clearly different values were obtained between male and female groups. Furthermore, increased values were seen in accordance with aging in all except 3 cases. Even this aging effect showed remarkable individual differences, which, however, lead to a better correlation to aging than increase of gamma-globulin. In many cases with ESR values out of +/- 2 SD of their mean values, their other laboratory findings such as general peripheral blood tests and plasma protein tests etc., showed also the results out of +/- 2 SD of their mean values. These results show that ESR can be a useful tool as one of screening parameters.

[Effect of monoamine oxidase inhibitors on the concentrations of free and conjugated dopamine in rat small intestine and liver following exogenous dopamine administration].

In rats treated with MAO inhibitors, free and conjugated catecholamine concentrations in small intestine and liver following exogenous free dopamine (DA) administration at the rate of 2 micrograms.kg-1.min-1 for two hours were measured. Free and conjugated DA concentrations increased in both small intestine and liver following exogenous free DA administration. Free and conjugated DA concentrations of the tissue of the rat treated with MAO inhibitors were higher than that of the control group. These results suggest that transformation of free DA into conjugated DA is activated in the small intestine and liver of the rat treated with MAO inhibitors when exogenous DA is given.

[Free and conjugated catecholamines in gastric and bile juice during surgical operations].

From 40 patients under surgical operation, gastric or bile juice was obtained to determine the possible excretion of free and conjugated catecholamine (CA) into the two kinds of juice. The patients were divided into normal renal function group and chronic renal failure group and former group was further divided into the dopamine (DA)-administered group and the non-DA-administered group. In the non-DA group with normal renal function, the gastric juice contained 0.30 +/- 0.11ng.ml-1 of free and 0.12 +/- 0.06ng.ml-1 of conjugated norepinephrine (NE) and also 0.09 +/- 0.07ng.ml-1 of free and 0.40 +/- 0.10ng.ml-1 of conjugated DA. In DA group, the gastric juice contained 2.01 +/- 0.41ng.ml-1 of free and 3.66 +/- 0.84ng.ml-1 of conjugated DA respectively. Significant differences in DA were observed between two groups. In the bile juice, the conjugated NE increased significantly from 0.15 +/- 0.05 ng.ml-1 to 1.24 +/- 0.34ng.ml-1 and also the conjugated DA increased significantly from 2.17 +/- 0.77ng.ml-1 to 21.33 +/- 5.23ng.ml-1 by infusion of DA at the rate of 2 micrograms.kg-1.min-1 for 197 +/- 48min. In chronic renal failure group, the conjugated NE and DA increased significantly to 1.04 +/- 0.27ng.ml-1 and 1.64 +/- 0.61ng.ml-1 respectively compared to that of normal renal group. It was confirmed that gastric juice and bile juice contain the free and conjugated CA during surgical operation and by the infusion of DA, free and conjugated DA are excreted into gastric juice and bile juice and also conjugated CA in the gastric juice from chronic renal failure group is increased.

[Adrenal catecholamine releases during hypotension with continuous administration of calcium entry blockers and nitroglycerin].

We investigated the effect of nicardipine and diltiazem on adrenal catecholamine releases, and compared it with the effect of nitroglycerin in dogs. A 35% reduction in mean arterial pressure was achieved and maintained for 45 minutes. The dogs were observed until 60 minutes after the discontinuation of drug infusions. Adrenal catecholamine releases increased during hypotension with continuous administration of nicardipine, diltiazem and nitroglycerin, but they showed no significant differences between each drug. There was no more increase in adrenal catecholamine releases in spite of additional nicardipine administration. These results suggest that Ca2+ antagonist such as nicardipine directly inhibits the catecholamine releases from the adrenal glands.

[Evaluation of efficiency of ACD-CPR and STD-CPR; a multi-institutional study].

We compared the efficacy of ACD-CPR and STD-CPR based on 64 multi-institutional reports. No significant differences were observed in the rate of restoration of spontaneous circulation (ROSC) and in cardiopulmonary parameters during CPR using the two methods. There were 5 cases in which cardiopulmonary parameters improved after switching from STD-CPR to ACD-CPR and, eventually, in two of them spontaneous circulation was restored. In the ROSC cases of both groups, ETCO2 and values of SpO2, PaO2, and systolic BP at 30 minutes were higher than those of non-ROSC cases. ETCO2 never exceeded 20 mmHg in the non-ROSC cases, but it was higher in the ROSC cases. ACD-CPR is a good choice when trained persons are present or when extra hands are available to continue the CPR.

Histochemical demonstration of free radicals (H2O2) in ischemic brain edema and protective effects of human recombinant superoxide dismutase on ischemic neuronal damage.

A new histofluorescence method by HPAA (p-hydroxyphenyl acetic acid) for free radicals in the brain tissue was devised to study neuronal damage induced by ischemia. Cerebral ischemia was produced in rats by injection of plastic microspheres and arachidonic acid (AA) into the right carotid artery. The concentration of malondialdehyde (MDA; free radical) in cerebral cortex of aminotriazol (an H2O2-dependent inhibitor of catalase) treated rats 2 h after stroke was 6.33 times the level before infarction, while the concentration of MDA in h-r SOD (free radical-scavenging enzyme) treated rats 2 h after stroke was significantly lower than in untreated rats. The histochemical findings demonstrated marked H2O2 production around blood vessels occluded by microspheres in the cerebral cortex of the aminotriazole treated rats 2 h after stroke together with disruption of the BBB. Light microscopical findings demonstrated extensive edematous changes in the aminotriazole treated rats 2 h after stroke, while pathological damage in SOD treated rat brains was absent or minimal. We conclude that free radicals are formed during ischemia, and that AA appears to be a major source of activated oxygen radicals. The findings indicate that SOD is protective against ischemia-induced neuronal damage.

Ischemia and reperfusion enhance ATF-2 and c-Jun binding to cAMP response elements and to an AP-1 binding site from the c-jun promoter.

The transcription factors controlling the complex genetic response to ischemia and their modes of regulation are poorly understood. We found that ATF-2 and c-Jun DNA binding activity is markedly enhanced in post-ischemic kidney or in LLC-PK1 renal tubular epithelial cells exposed to reversible ATP depletion. After 40 min of renal ischemia followed by reperfusion for as little as 5 min, binding of ATF-2 and c-Jun, but not ATF-3 or CREB (cAMP response element binding protein), to oligonucleotides containing either an ATF/cAMP response element (ATF/CRE) or the jun2TRE from the c-jun promoter, was significantly increased. Binding to jun2TRE and ATF/CRE oligonucleotides occurred with an identical time course. In contrast, nuclear protein binding to an oligonucleotide containing a canonical AP-1 element was not detected until 40 min of reperfusion, and although c-Jun was present in the complex, ATF-2 was not. Incubating nuclear extracts from reperfused kidney with protein phosphatase 2A markedly reduced binding to both the ATF/CRE and jun2TRE oligonucleotides, compatible with regulation by an ATF-2 kinase. An ATF-2 kinase, which phosphorylated both the transactivation and DNA binding domains of ATF-2, was activated by reversible ATP depletion. This kinase coeluted on Mono Q column chromatography with a c-Jun amino-terminal kinase and with the peak of stress-activated protein kinase, but not p38, immunoreactivity. In conclusion, DNA binding activity of ATF-2 directed at both ATF/CRE and jun2TRE motifs is modulated in response to the extreme cellular stress of ischemia and reperfusion or reversible ATP depletion. Phosphorylation-dependent activation of the DNA binding activity of ATF-2, which appears to be regulated by the stress-activated protein kinases, may play an important role in the earliest stages of the genetic response to ischemia/reperfusion by targeting ATF-2 and c-Jun to specific promoters, including the c-jun promoter and those containing ATF/CREs.