Influence of single and concurrent clofibrate and phenobarbital administration on cytochrome P450-dependent mixed function oxidase activities and peroxisome proliferation in male rat liver. The influence of both single and concurrent administration of phenobarbital and clofibrate on hepatomegaly, cytochrome P450-dependent mixed function oxidase activities, and peroxisome proliferation in male rat liver have been studied. Both xenobiotics separately increase the liver: body weight ratio and their combined administration results in greater hepatomegaly than either compound alone. Both compounds induce NADPH-cytochrome c(P450) reductase activity and laurate omega- and omega-1-hydroxylase activities, but only phenobarbital induces pentoxyresorufin-O-dealkylase. None of the drug treatments induced microsomal cytochrome b5. Phenobarbital did not cause peroxisome proliferation and inhibited the corresponding clofibrate-dependent proliferation. Taken collectively, our studies have demonstrated that concomitant treatment with phenobarbital and clofibrate are largely permissive with respect to the hepatic mixed function oxidase system but have opposing effects on the phenomenon of peroxisome proliferation in the same tissue.


A genetic study of type 2 neurofibromatosis in the United Kingdom. I. Prevalence, mutation rate, fitness, and confirmation of maternal transmission effect on severity. A clinical and genetic study of type 2 neurofibromatosis (NF2) has been carried out in the United Kingdom. Virtually complete ascertainment of cases in the north-west of England was achieved and suggests a population incidence of 1 in 33,000 to 40,000. In the UK as a whole, 150 cases have been identified and been used to study the clinical and genetic features of NF2. The autosomal dominant inheritance of NF2 was confirmed, 49% of cases were assessed as representing new mutations, and the mutation rate was estimated to be 6.5 x 10(-6). Evidence to support a maternal gene effect was found in that age at onset was 18.17 years in 36 maternally inherited cases and 24.5 in 20 paternally inherited cases (p = 0.027). The preponderance of maternally inherited cases was also significant (p = 0.03). Data are presented which suggest that there are two types of NF2, one with later onset and bilateral vestibular schwannomas as the only usual feature, and the other with earlier onset and multiple other tumours. A considerable number of cases did not fall easily into one or other group and other factors such as maternal effect on severity and anticipation need to be considered.


A genetic study of type 2 neurofibromatosis in the United Kingdom. II. Guidelines for genetic counselling. The major defining features, age at onset of symptoms, and survival in 150 patients with type 2 neurofibromatosis (NF2) have been studied. The mean age at onset was 21.57 years (n = 110) and no cases presented after 55 years of age. Patients presented with symptoms attributable to vestibular schwannomas (acoustic neuroma), cranial meningiomas, and spinal tumours. In 97 cases studied personally by the authors, skin and eye examination were found to be useful to detect early signs of the condition. Examination of the skin is likely to assist in early diagnosis in at least 10% of cases and examination of the eye for a lens opacity or cataract in at least as many again. There are marked interfamilial differences in disease severity and tumour susceptibility. Vestibular schwannomas are not fully penetrant, but the condition is usually expressed in another way. Alteration to the current diagnostic criteria is advocated to cover the lack of provision for new mutations. A screening protocol is proposed and the effect of disease heterogeneity on management is discussed.


Linkage investigation of three putative tuberous sclerosis determining loci on chromosomes 9q, 11q, and 12q. The Tuberous Sclerosis Collaborative Group. Previous linkage studies in tuberous sclerosis have implicated three disease determining loci at 9q, 11q, and 12q. We have collated phenotypic and genotypic data on 1622 members of 128 families with tuberous sclerosis in order to evaluate simultaneously the evidence for these putative loci. Affection status in the family members has been reassessed using uniform diagnostic criteria and genotypic data extensively checked before analysis under alternative models of locus heterogeneity. One tuberous sclerosis determining locus, accounting for approximately 50% of the families studied, has been found to map in the region of D9S10 on 9q34 but no evidence has been found to support the existence of major loci on 11q or 12q. A locus, or loci, elsewhere in the genome is likely to account for tuberous sclerosis in most non-chromosome 9 linked families.


Computer simulation of linkage and heterogeneity in tuberous sclerosis: a critical evaluation of the collaborative family data. The existence of locus heterogeneity for a genetic disease may complicate linkage studies considerably, especially when very few large families with the disease are available. In this situation a modest collection of families is unlikely to be sufficient for successful localisation of one or more disease genes. Recently, eight research groups working on tuberous sclerosis (TSC) brought together linkage data pertaining to the candidate chromosomes 9, 11, and 12 for a large group of families. In a series of simulation studies we determined the probability of detecting linkage and linkage heterogeneity in this set of families. On average TSC families are very small; in most cases there are fewer than two informative meioses. The size distribution of chromosome 9 linked families was similar to that of non-linked families. This indicates that a dramatic difference in the clinical severity of major genetic forms of TSC is unlikely. The results of our simulation studies show that this set of families can generate highly significant evidence for linkage and heterogeneity. When two TSC genes are equally common, the strongest evidence for linkage and heterogeneity could be obtained using a method based on the incorporation of multiple candidate regions in a single analysis, with an average lod score of 24.27.


Deficiencies of fibrillin and decorin in fibroblast cultures of a patient with neonatal Marfan syndrome. Changes in the structure and metabolism of fibrillin, a microfibril associated protein, can result in classical Marfan syndrome, and reduced expression of decorin, a small extracellular chondroitin sulphate/dermatan sulphate proteoglycan, has been observed in fibroblasts of a patient with neonatally lethal Marfan syndrome. We have studied the synthesis of fibrillin and decorin in cultured fibroblasts of a further sporadic patient with neonatally lethal Marfan syndrome. Fibrillin immunoreactivity in the extracellular matrix of the patient's fibroblasts was markedly reduced, and the fibrillar pattern was absent, in spite of normal amounts of fibrillin mRNA. Decorin mRNA, synthesis, and immunoreactivity in the matrix were also reduced. The results indicate involvement of both fibrillin and decorin in the pathogenesis of neonatal Marfan syndrome in this patient, but do not indicate which is the primary defect. We speculate, however, that a structural defect of fibrillin leads to diminished incorporation of the protein into the extracellular matrix, and that underexpression of decorin is secondary to the primary fibrillin defect. Combined deficiency of fibrillin and decorin may be the cause of the severe clinical phenotype.


Severity of chest disease in cystic fibrosis patients in relation to their genotypes. A detailed comparison of the severity of chest disease with mutational status was carried out by cross sectional study of 127 cystic fibrosis patients, aged 1 to 31 years, living in Wales. Lung disease was classified according to severity, depending on pulmonary function tests (carried out on 76 patients) and chest radiograph status; information was obtained also on age at diagnosis in relation to severity of chest disease and colonisation with Pseudomonas species. Genotypes were determined by analysis for the mutations delta F508, delta I507, G551D, R553X, G542X, R117H, R560T, 1717--IG > A, and 621 + 1G > T. CF patients homozygous positive and heterozygous for the delta F508 deletion showed a significant decline of lung function with age. Unlike other studies, we did not find patients homozygous positive for the delta F508 deletion to have poorer lung function compared with heterozygous patients. Patients with the genotype 621 + IG > T/delta F508 tended to have more severe chest disease than the delta F508 homozygous patients in the same age group. There was some evidence that four patients heterozygous for R117H have mild chest disease.


Predicted and observed sizes of dystrophin in some patients with gene deletions that disrupt the open reading frame. Among 85 patients with Duchenne and Becker muscular dystrophy, 29 were found to have mutations which disrupted the open reading frame for dystrophin. Thus any dystrophin detected in this group of patients should consist of the severely truncated polypeptides that represent prematurely terminated translation products. Dystrophin was detected in blots from 17/29 biopsies and the observed sizes of the polypeptides were compared with predicted sizes calculated in two ways: if translation was terminated at the stop codon generated by each frameshifting deletion, and if the reading frame was restored and translation proceeded. In every case the observed size matched the size predicted on the basis of a restored reading frame. This was in accord with immunocytochemical labelling of scattered dystrophin positive fibres which were found on serial sections labelled with antibodies to both the rod and C-terminal domains. Thus analysis at the protein level supports genetic evidence of exon skipping as a mechanism which restores frameshifting mutations in some fibres.


Nine generations of a family with autosomal dominant retinitis pigmentosa and evidence of variable expressivity from census records. We present a nine generation family with autosomal dominant retinitis pigmentosa (ADRP). Evidence of blindness in the early generations, as obtained from census returns and clinical records, and examination of current patients show variable expressivity with a spectrum which ranges from asymptomatic in late life to blindness in the third decade of life. The family is not linked to any of the chromosomal locations so far described in ADRP and further illustrates the heterogeneity of the disorder.


Causes of death in patients with Huntington's disease and in unaffected first degree relatives. Causes of death were examined from death certificates for 395 Danish subjects with Huntington's disease (HD) and for 282 unaffected sibs and compared with the causes of death in the general Danish population. For both the HD subjects and the sibs, pneumonia and cardiovascular diseases were the most frequent primary causes of death. Suicides accounted for 5.6% of all deaths among the HD subjects and, unexpectedly, for 5.3% among the sibs, some of whom may have been carriers of the HD gene. Both were significantly higher than the corresponding frequency of 2.7% in the general Danish population, but there was no evidence of differences in the age specific proportions for the HD subjects and for the sibs compared to the general population. Some accidents leading to death in the sibs may have been hidden suicides. The rate of cancer was low for the HD patients, being only 5.3% compared with 31.2% for the sibs. Neurological diseases were reported with an increased frequency in the HD patients compared to the general population, 6% v 1%.


The response to ischemia in blood perfused vs. crystalloid perfused isolated rat heart preparations. With a research hypothesis that the behavior of blood perfused hearts was different from that of crystalloid perfused hearts, we tested the null hypothesis that the functional and metabolic status of blood-perfused (paracorporeal oxygenation) and Krebs-Henseleit (bubble oxygenation) perfused Langendorff isolated rat hearts is the same before, during and after global myocardial ischemia. Thirty isolated rat hearts were studied under identical conditions except that in equal numbers they were randomly assigned to either blood or crystalloid perfusion. In the blood perfused and crystalloid perfused hearts subjected to 22 min of normothermic ischemia and 30 min of reperfusion, mean systolic recovery was 72 +/- 3.9% (S.E.) and 20 +/- 10% (P = 0.001), respectively; coronary resistance increased 21 +/- 16% and 158 +/- 27% (P = 0.0003) (unadjusted for viscosity); mean water content after reperfusion was 82.0 +/- 0.43% and 86.7 +/- 0.42% (P < 0.0001), ATP content was 8.4 +/- 1.9 and 4.3 +/- 0.5 mumol/g dry wt (P = 0.08), and energy charge was 0.74 +/- 0.114 and 0.59 +/- 0.048 (P = 0.3). A major qualitative difference during reperfusion was spontaneous relaxation of contracture and rapid resumption of sinus rhythm in blood perfused hearts, in contrast to continued contracture and rise in intraventricular pressure in 9 of 10 crystalloid perfused hearts. One crystalloid perfused heart did not develop contracture, and its phenomena during reperfusion were similar to those of blood perfused hearts. The data support the research hypothesis, and suggest caution in extrapolating to blood perfused systems inferences from crystalloid perfused models. Better preservation of reactive hyperemia early in reperfusion may explain the better performance of blood perfused hearts.


Myocardial contractility, lusitropy and calcium responsiveness in young (50 days) and hypertrophied (180 days) cardiomyopathic hamsters. Contractility, lusitropy and responsiveness to the increase of external Ca2+ concentration were studied in left ventricular papillary muscles of normal and cardiomyopathic Syrian hamsters (SCH) from the UM-X 7.1 strain, both at the onset of myolysis (50-day-old animals) and at the cardiac hypertrophy stage (180-day-old animals) in the absence of congestive heart failure. A marked decrease in all indices of systolic performance was observed in 180-day-old myopathic hamsters as compared to age-matched controls. This was associated with (1) an impairment of the relaxation phase, (2) a loss of the load sensitivity of relaxation, and (3) a decrease in the inotropic and lusitropic responsiveness to Ca2+. On the other hand, when some indices of contraction and the inotropic response to Ca2+ were impaired in 50-day-old myopathic hamsters as compared to age-matched controls, relaxation phase and the lusitropic response to Ca2+ did not alter. This study shows that, in the UM-X 7.1 myopathic hamsters at the earlier stage of the disease, alterations in calcium homeostasis and contraction seem to be the first determinant factors of the development of heart failure when relaxation is not impaired. Conversely, when cardiac hypertrophy has developed, impaired relaxation may worsen heart failure.


Metabolism of platelet-activating factor in the guinea-pig heart. Platelet-activating factor (PAF; 1-alkyl-2-acetyl-glycero-phosphocholine) is a biologically active phospholipid which is synthesized by a variety of blood cells and organ systems. PAF exerts many effects on the cardiovascular system including hypotension, depression of myocardial contractility and coronary constriction. The present study has examined the capacity of the guinea-pig heart to regulate the levels of exogenous PAF in two different models: isolated perfused heart and isolated ventricular myocytes. In the first model, isolated hearts were perfused with labeled PAF (10(-10) M) in a recirculating manner at flow rates of 15 ml/min (normal flow perfusion; NFP) and 2 ml/min (low flow perfusion, LFP). Exogenously provided PAF appeared in the tissue in a time-dependent manner. The rate of extraction of PAF was higher during LFP than during NFP. PAF was metabolized by the heart to two major products, lyso-PAF and 1-alkyl-2-acyl-sn-glycero-3-phosphocholine (1-alkyl-2-acyl-GPC). Lyso-PAF was found primarily in the perfusion buffer while both lyso-PAF and 1-alkyl-2-acyl-GPC were detected in the tissue. No qualitative difference in the metabolic products derived from PAF catabolism was observed between hearts undergoing NFP and LFP. Acetyl hydrolase activity was detected in the perfusion fluid at both flow rates, probably accounting for the formation of lyso-PAF in the perfusate. However, perfusion fluid from LFP contained a higher acetyl hydrolase activity, per micrograms of protein as compared to fluid from NFP. Isolated ventricular myocytes incubated with labeled PAF (3 x 10(-9) M) also converted it to 1-alkyl-2-acyl-GPC. Kinetic experiments suggested that PAF was initially deacetylated to form lyso-PAF and that this intermediate was then rapidly reacylated with a fatty acyl moiety at the sn-2 position. HPLC analysis of the fatty acids inserted at the sn-2 position of 1-alkyl-2-acyl-GPC revealed that the myocytes reacylated lyso-PAF predominantly with arachidonic acid. These data indicate that the guinea-pig heart may regulate PAF levels by at least two mechanisms: (1) it may release acetyl hydrolase into the vascular compartment, particularly under low flow conditions; and (2) the ventricular myocyte has the capacity to take up PAF and catabolize it to inactive products.


Ischaemic preconditioning and contractile function: studies with normothermic and hypothermic global ischaemia. A significant reduction in the extent of cell necrosis or the incidence of reperfusion-induced arrhythmias can be achieved with ischaemic preconditioning. If preconditioning was also found to be effective in protecting against global ischaemia, then this may have significant implications for the preservation of the heart during cardiac surgery. We therefore investigated this phenomenon in relation to recovery of contractile function after global ischaemia in the isolated rat heart. Isolated working rat hearts (n = 6 per group) were perfused aerobically at 37 degrees C for 20 min and contractile function recorded. This was followed by 10 min of aerobic Langendorff perfusion (control hearts) or 5 min global ischaemia (37 degrees C) + 5 min Langendorff reperfusion (preconditioned hearts). The hearts were then subjected to 10, 15, 20 or 25 min of global ischaemia (37 degrees C) and reperfusion (15 min Langendorff + 20 min working) after which function was again assessed. Preconditioning improved functional recovery after all durations of ischaemia. Thus aortic flow after 10, 15, 20 and 25 min of ischaemia and 35 min of reperfusion recovered to 84, 58, 16 and 5%, respectively, in controls and 88, 74, 55 and 20%, respectively, in the preconditioned groups. To assess whether preconditioning was effective in a surgically relevant model of hypothermic ischaemia, the experiments were repeated with longer periods (45, 70, 90, 115, 135 and 160 min) of ischaemia at 20 degrees C. Under these conditions, normothermic preconditioning increase the post-ischaemic recovery of aortic flow after 115, 135 and 160 min of ischaemic (from 36, 20 and 10%, respectively, in controls to 57, 39 and 26%, respectively, in preconditioned hearts). There was no consistent correlation between tissue high energy phosphate content and enhanced post-ischaemic recovery. Thus, we have demonstrated that ischaemic preconditioning can improve contractile function after global ischaemia in the isolated rat heart, we have defined the duration of ischaemia for which it is operative, and we have shown that this protection is additive to that of hypothermia-induced protection during ischaemia. This may have clinical implications for cardiac surgery.


Correlation of contractile dysfunction and abnormal tissue energy metabolism during hypoperfusion with norepinephrine in isolated rat hearts: differences between normal and diabetic hearts. The relationship of myocardial high-energy phosphate depletion and lactate accumulation with contractile dysfunctions was investigated in streptozotocin-diabetic (DM) and normal rat hearts. The isolated hearts were perfused with 10(-6) M norepinephrine (NE) at various low-flow rates (0.4-6 ml/min/g heart wt) for 1 h. Left ventricular pressure (LVP) and contractile force (CF) were monitored, through a water-filled balloon in LV and through a hook attached to the apex, respectively. In DM hearts resting CF (diastolic tension) increased, when the perfusion flow rate was reduced below 6 ml and reached a maximum at a flow rate of less than 3 ml. The large increase in LV stiffness correlated with an elevation in diastolic LVP. In normal hearts these parameters were elevated at a flow rate below 1 ml. A flow-dependent decrease in developed CF was more prominent in DM than in normal hearts, while developed LVP and perfusion pressure were slightly higher in DM hearts with a marked increase in the LV stiffness. A flow-dependent decrease in high-energy phosphates and increases in inorganic phosphate and lactate were more prominent in the inner than in the outer layer of LV free wall in both groups. The change of ATP in the inner layer was greater, while increases of lactate in both layers were smaller in DM hearts. Changes in mechanical parameters correlated well with the ATP decrease and lactate increase in the inner layer in both groups. The correlation curves, however, were not coincidental: at the same low ATP and high lactate level, the LV stiffness was higher in DM hearts. Results indicate that DM hearts are more susceptible to flow-reduction with NE and depletion of total ATP in their tissue, and easily suffer from increased LV stiffness. This cannot be explained by the rate of decrease in total ATP and lactate accumulation alone.


Developmental changes in tolerance to ischaemia in the rabbit heart: disparity between interpretations of structural, enzymatic and functional indices of injury. The vulnerability of the heart to injury during ischaemia and reperfusion and its responsiveness to various protective and pharmacological interventions are age-dependent. Using three independent indices of tissue injury (cardiac structure, contractile function and creatine kinase leakage), we compared the response of adult (60-90 days old) and neonatal (7 days old) isolated perfused rabbit hearts to global ischaemia and reperfusion. Prior to ischaemia, heart rate was significantly higher in neonatal hearts, as were control values for coronary flow, aortic flow and cardiac output when expressed on a dry wt basis. In experiments in which adult and neonatal hearts (n = 8 per group) were subjected to 2 min of cardioplegia and 45 min of ischaemia, the post-ischaemic recovery of all indices of cardiac function (when expressed as a percentage of pre-ischaemic control) was significantly higher in neonatal than in adult hearts. Thus, cardiac output recovered to 82.9 +/- 3.6% in the neonate but to only 57.9 +/- 6.7% in the adult (P < 0.05). The functional evidence of a greater resistance to ischaemia in the neonate was, however, contradicted by the levels of creatine kinase leakage which tended to be greater in the neonatal than in the adult heart (32.0 +/- 4.7 vs 20.0 +/- 3.1 IU/15 min/g dry wt). Morphological studies indicated that injury was comparable (moderate-to-severe in degree) in both groups. To assess further the relationship between the three indices, additional experiments were undertaken in which the duration of ischaemia in the neonate was extended to 60 min so that the post-ischaemic recovery of function was reduced to a level similar to that seen in the adult after 45 min of ischaemia. Under these conditions cardiac output recovered to 55.6 +/- 4.8% in the neonatal heart (P = NS when compared with the adult) and creatine kinase leakage increased to 88.2 +/- 13.9 IU/15 min/g dry wt--a value over four times greater than that measured in adult hearts with a comparable degree of functional injury. Morphological examination of tissue obtained after 15 min of reperfusion revealed a remarkable recovery of structure in both age groups. In conclusion, in functional terms the neonatal heart was more resistant to ischaemia than the adult; enzymic leakage, however, indicated the opposite and structural assessment revealed no differences. Thus, in comparing injury during ischaemia and reperfusion between different age groups, it is clearly important to employ several independent indices.


Diffusion distances, total capillary length and mitochondrial volume in pressure-overload myocardial hypertrophy. We examined the relationships between blood pressure, coronary blood flow, cardiac output, myofiber growth, capillarity, mitochondrial content, and capillary and mitochondrial distributions in a pressure-overload model of myocardial hypertrophy. The Goldblatt one kidney-one clip (1K1C) procedure was performed on seven adult rabbits. After 1 month, mean blood pressure increased 50% and mean heart mass increased 30%. Coronary blood flow and cardiac output at rest were similar in control and 1K1C hearts; cardiac output fell 40% when 1K1C hearts were paced to 35% above basal heart rate. Capillary density in the left ventricular free wall (LV) decreased with increasing fiber size by as much as 30%. However, capillary-to-fiber ratio and total capillary length in the LV increased with heart size by up to 30% and 80%, respectively. This indicated that there was some proliferation of capillaries taking place, but not enough in comparison to fiber growth to prevent the lengthening of distances between capillaries. Mitochondrial volume density decreased by as much as 30% with increasing heart size, but total mitochondrial volume increased up to 80%. This indicated that there was some proliferation of mitochondria, but not enough to prevent dilution of mitochondria by the growing myofibrillar elements. Analysis of the distribution of mitochondria suggested that the new mitochondrial material was added to the center of myofibers, thereby further lengthening oxygen diffusion distances. There was a constant ratio of 10.4 +/- 0.3 km of capillaries per ml of mitochondria in 1K1C and control hearts, demonstrating that the structures for oxygen supply and consumption were remaining in fixed proportion to each other. There was no evidence that the decreased performance of paced 1K1C hearts was attributable to an oxygen diffusion limitation to mitochondria.


Developmental differences in the response of cytosolic free calcium to potassium depolarization and cardioplegia in cardiac myocytes. Several biochemical and functional characteristics of immature myocardium suggest a diminished capacity to regulate intracellular Ca2+ during stress. In particular, cellular calcium overload has been postulated as an important pathogenetic mechanism accounting for suboptimal functional recovery following cardioplegia in immature myocardium. Using intracellular Fura-2 fluorescence as Ca2+ indicator, we measured cytosolic free calcium ([Cai]) in single myocytes and cell suspensions derived from both juvenile (4 weeks post-partum) and mature (6-12 months post-partum) New Zealand white rabbits. Resting [Cai] in juvenile heart cells (26 +/- 3 nM) were approximately 50% of that found in adult myocytes (55 +/- 5 nM). In addition, on exposure to increasing concentrations of extracellular potassium ([Kex]), adult but not juvenile myocytes exhibited increases in [Cai]. These two observations underscore developmental differences in intracellular Ca2+ homeostasis. Of particular clinical relevance is the [Cai] response to cardioplegia containing 16 mM [Kex]: neither group demonstrated the expected [Cai] increase in response to potassium depolarization. The lack of [Cai] response to cardioplegia was most likely due to the high levels of Mg2+ (32 mM) contained in cardioplegic solutions. We conclude that cellular calcium overload does not occur following exposure to cardioplegia alone. Accordingly, these findings do not account for recognized developmental differences in functional recovery from "myocardial protection".


Analysis of the upstream regulatory region of human ventricular myosin light chain 1 gene. To explore the mechanisms regulating expression of ventricular myosin light chain 1, the human gene including 5'-flanking DNA was cloned and characterized by Southern blot and restriction mapping. A 2 kb 5'-flanking DNA was sequenced and linked to a chloramphenicol acetyltransferase reporter gene. The constructs then were transfected into cultured human and rat cardiomyocytes as well as rat aortic endothelial cells. Deletion analysis of constructs revealed that the basal promoter sequences, which were located within 62 base pairs of the cap site, could direct high levels of chloramphenicol acetyltransferase gene expression in the cardiomyocytes and endothelial cells. The region between -62 to -312 base pairs strongly repressed the chloramphenicol acetyltransferase gene expression in the cardiomyocytes and endothelial cells. Positive elements were found between -312 and -2000 base pairs of the cap site. These results are indicative, among other possibilities, that the human ventricular myosin light chain 1 gene is turned on in cardiomyocytes by the presence of trans-acting factors that are bound to upstream positive elements and is turned off in non-muscle cells by the presence of repressor-binding proteins. But this mechanism remains to be established.


Energy metabolism in myocardial stunning. We investigated the effect of reversible ischemia, leading to persistent contractile dysfunction (stunning), on myocardial energy metabolism. The balance of energy metabolism is expressed by the phosphorylation state of cytosolic nucleotides. This variable cannot be measured directly because of nucleotide compartmentation, but in the isolated heart it can be estimated by the release of purine catabolites. We have previously shown that increased energy consumption or impaired energy production cause purine release to increase, while primary reduction in energy consumption has the opposite effect. Isolated working rat hearts were reperfused after 10 min of global ischemia, measuring hemodynamic variables, tissue high energy phosphate compounds and purine release. In post-ischemic recovery, aortic flow and minute work decreased to 82 +/- 3% and 77 +/- 4% of control, adenine nucleotide pool was reduced by 4.6 mumol/g dry wt, phosphocreatine to creatine ratio increased significantly and purine release decreased to 42 +/- 6% (P < 0.01). The rate of purine salvage, as evaluated by the incorporation of exogenous 3H-adenosine and 14C-hypoxanthine into tissue nucleotides, was much lower than net purine release, and was unchanged after ischemia and reperfusion. The adenine nucleotide pool could be depleted to the same extent as in the stunned myocardium by prolonged (60 min) aerobic perfusion. In this group the hemodynamic variables were unchanged and purine release averaged 87 +/- 9% of control (P = NS). In other experiments prolonged perfusion was combined with preload reduction in order to decrease energy demand. This protocol reproduced the effects of ischemia-reperfusion: aortic flow and minute work averaged 79 +/- 4% and 73 +/- 9% of control, adenine nucleotide depletion was 4.4 mumol/g dry wt and purine release decreased to 38 +/- 5% (P < 0.01). Our findings support the view that stunning is not due to adenine nucleotide depletion or to impairment in energy production, which would cause purine release to increase, but rather to primary reduction in energy utilization.


Taurine antagonizes the increase in intracellular calcium concentration induced by alpha-adrenergic stimulation in freshly isolated guinea-pig cardiomyocytes. The effect of taurine on phenylephrine alpha-adrenergic action was studied in freshly-isolated guinea-pig ventricular myocytes. Intracellular calcium concentration was measured at nearly physiological extracellular CaCl2 in both cell suspension using quin-2 (mean intracellular calcium concentration 154.0 +/- 8.0 nM, n = 24), and single myocyte with fura-2 (mean intracellular calcium concentration 159.0 +/- 20.0 nM, n = 23). Phenylephrine increased intracellular calcium concentration in both preparations. In cell suspensions in the presence of 10(-6) M propranolol and at 2.2 mM extracellular calcium concentration, phenylephrine dose-dependently (3 x 10(-7)-10(-5) M) increased intracellular calcium concentration, its effect being abolished in the presence of phentolamine. Taurine 20 mM in the incubation fluid increased taurine content in the cells from 110 +/- 7 nmol/mg to 317 +/- 49 nmol/mg of total proteins. In the range 0.5-20 mM, taurine concentration-dependently reduced the phenylephrine-induced intracellular calcium increase in cell suspensions and 20 mM taurine decreased the effect of 10(-5) M phenylephrine (measured in the presence of 10(-6) M propranolol) by about 80%. Beta-Alanine (20 mM) did not modify the phenylephrine effect. Our data show that the "protective" effect of taurine in in vitro cardiac preparations and in cardiomyocyte isolation procedures is due to its effect on cardiomyocyte intracellular calcium ion concentration, and that taurine specifically antagonizes alpha-adrenoceptor activation.


Energetic determinants of stunning and cell damage following reoxygenation of rabbit myocardium. The energetic conditions in anoxia which lead to stunning and cell damage upon reoxygenation of the myocardium are studied in isolated papillary muscles of rabbit contracting isometrically at 20 degrees C. Before anoxia and thereafter the muscles are stimulated at 0.2 Hz, while during anoxia stimulation frequency is varied. Total creatine (CrT) content is taken as a measure of cell damage. The degree of stunning is estimated from the difference in force before anoxia and at the end of the recovery period when no creatine is lost. In anoxic rabbit papillary muscles glycogen is the substrate for lactate formation. The 'ATP reserve' of the muscle can then be calculated from: 2 ATP + PCr + ADP + 3 (glycosyl units). In control muscles it equals 482.2 mumol.g-1 dw of ATP. By varying stimulus frequency (0, 0.2, 0.5, or 1 Hz) during and the length of the anoxic period (20 to 80 min), the fraction of the ATP reserve used in anoxia is varied. It is found that the degree of stunning depends on the degree of ATP reserve depletion. When the ATP reserve becomes virtually zero force recovers to only 67% of the preanoxic value. When anoxia continues after the ATP reserve is virtually zero, creatine in the muscle decreases during reoxygenation, indicating cell damage. Apparently a hierarchy of cellular ATPases exists in the myocardium when energy is short preserving cell integrity as long as possible.


Endothelin release during ischaemia and reperfusion of isolated perfused rat hearts. The hypothesis tested was that release of endogenous endothelin plays a role in events associated with or leading to myocardial ischaemia and/or post-ischaemic reperfusion damage. Release of endogenous endothelin into the coronary perfusate of isolated perfused rat hearts during ischaemia and reperfusion was measured with a sensitive radioimmunoassay using a polyclonal antibody with 100% cross-reactivity for all three endothelin isomers. Basal endothelin release was 0.69 +/- 0.02 pg/min/g wet heart weight (n = 35) and was constant up to 180 min. During low-flow hypoxic ischaemia for 180 min (PO2 approximately 250 mmHg) and in the presence of 1% foetal calf serum, the release rate was reduced to below 10% of controls (P < 0.01) and increased four-fold on reperfusion (P = 0.05). The influence of endothelin on vascular and myocardial reperfusion damage was studied with exogenous endothelin-2. After 1 h of low-flow ischaemia, endothelin-2 increased the coronary perfusion pressure to a similar extent as in non-ischaemic hearts, but with a 30-times higher potency. The threshold dose for the constrictive effect was approximately 100 to 300 pg per heart, about ten times more than was recovered in the coronary effluent upon reperfusion. The influence of endothelin on myocardial reperfusion mechanical function (stunning) was assessed with 100 ng endothelin-2, a dose some 3500-fold higher than the amount released during 30 min reperfusion. This dose, given at the onset of reperfusion, improved post-ischaemic aortic output recovery during the first 20 min of reperfusion, but worsened it thereafter (up to 40 min). These data indicate that, in the isolated perfused rat heart model, (1) endothelin is released in measurable amounts into the coronary circulation, (2) the release is much reduced during ischaemia and increased on early reperfusion following prolonged ischaemia, (3) based on the amounts released and the post-ischaemic sensitization of the coronary vasculature to endothelin, the peptide could contribute to reperfusion vascular damage, and (4) endothelin is unlikely to influence stunning owing to the extremely high dose necessary to alter reperfusion mechanical function.


Inhibition of adenosine metabolism increases myocardial interstitial adenosine concentrations and coronary flow. We employed an isolated guinea-pig heart model perfused at constant pressure (70 cmH2O) to test the hypothesis that inhibition of adenosine metabolism increases interstitial adenosine concentrations (as measured with epicardial discs) and coronary flow. Iodotubercidin (ITU, 1 microM) and EHNA (erythro-9-[2-hydroxy-3-nonyl] adenine, 5 microM) were used to inhibit adenosine kinase and deaminase, respectively during control conditions and during metabolic stimulation with 1 microM isoproterenol. The adenosine receptor blocker 8-phenyltheophylline (8-PT) was used during control conditions to assess whether the response seen was adenosine specific. ITU plus EHNA decreased heart rate (202 +/- 10 to 136 +/- 11 beats/min) and increased coronary flow (8.2 +/- 0.3 to 12.4 +/- 0.9 ml/min/g) without a change in MVO2, developed pressure or dP/dt. ITU plus EHNA increased adenosine concentrations in epicardial fluid (0.24 +/- 0.07 microM to 1.02 +/- 0.09 microM) and venous effluent (40 +/- 3 nM to 262 +/- 32 nM) during control conditions, and adenosine release increased from 389 +/- 96 pmols/min/g to 3480 +/- 365 pmols/min/g. 8-PT infusion reversed the effects on heart rate and coronary flow and resulted in a persistent elevation of epicardial fluid adenosine concentrations. During metabolic stimulation with 1 microM isoproterenol, ITU plus EHNA significantly limited the increase in heart rate and ventricular developed pressure and dP/dt while coronary flow increased to a significantly greater extent. Myocardial oxygen consumption was similar during metabolic stimulation between the two groups (vehicle vs. ITU plus EHNA). Epicardial fluid adenosine concentration in the vehicle-treated group increased from 0.17 +/- 0.3 microM to 0.34 +/- 0.02 microM at 15 min of isoproterenol stimulation whereas it increased from 1.10 +/- 0.02 microM to 2.90 +/- 0.46 microM in the ITU plus EHNA-treated group. Inhibition of adenosine metabolism during metabolic stimulation significantly increased venous adenosine concentrations and adenosine release and reduced inosine and hypoxanthine release proportionately. The release of adenosine+inosine+hypoxanthine was unchanged. Inhibition of adenosine metabolism provides evidence supporting the hypothesis that adenosine plays a role in regulating coronary vascular resistance as well as influencing heart rate and ventricular inotropy.


Mitochondrial abnormalities in myocardium of dogs with chronic heart failure. The number, size and structural integrity of mitochondria (MIT) were evaluated in the myocardium of 12 dogs with chronic heart failure (CHF) produced by sequential intracoronary microembolizations (EMB). Tissue specimens for transmission electron microscopy were obtained from the left ventricular (LV) free wall, septum and right ventricular free wall 3 to 4 months after the last EMB. Comparisons were made with samples obtained from identical sites in 9 control dogs. In dogs with CHF, LV ejection fraction decreased from 61 +/- 1% at baseline (prior to EMB) to 22 +/- 2% 3 to 4 months after the last EMB (P < 0.01) while plasma norepinephrine (PNE) concentration increased from 364 +/- 12 pg/ml to 837 +/- 150 pg/ml (P < 0.01). The number of MIT in an area of 100 square sarcomeres was greater in CHF dogs compared to controls (92 +/- 5 vs 64 +/- 2) (P < 0.001); whereas the average size of MIT was smaller (0.53 +/- 0.03 vs. 0.78 +/- 0.04 microm2) (P < 0.001). Injury ranging in severity from matrix depletion to myelinization and membrane disruption was present in 27 +/- 4% of MIT of CHF dogs compared to only 3 +/- 1% of MIT of controls (P < 0.001). MIT abnormalities were present to the same extent in all three regions of the heart. The severity of MIT injury, assessed on the basis of an injury index, was significantly higher in CHF dogs with PNE > or = 600 pg/ml (0.64 +/- 0.07) compared to CHF dogs with PNE < 600 pg/ml (0.32 +/- 0.08) (P < 0.01). Among CHF dogs, the MIT injury index was linearly related to PNE concentration (r = 0.57, P < 0.05), LV ejection fraction (r = 0.57, P < 0.05) and LV end-diastolic pressure (r = 0.57, P < 0.05). These data indicate that profound MIT abnormalities are present in the myocardium of dogs with CHF and are related to PNE concentration and to the severity of LV dysfunction.


Pyrimidine nucleotide synthesis is preferentially supplied by exogenous cytidine in adult rat cultured cardiomyocytes. The metabolism of pyrimidine nucleotides was studied in non-contracting myocytes isolated from adult rat hearts and compared to that observed in freshly prepared myocardium. The myocytes were cultured for up to 96 hrs in a commercial medium containing 50 microM cytidine, uridine, adenosine and adenine; 20 microM guanosine, thymidine and D-ribose; and 5 microM hypoxanthine, xanthine, guanine, thymine and uracil. Nucleotide pool sizes were measured by HPLC. Nucleotide and RNA labelling were followed by incorporation of [U-14C]-cytidine or [U-14C]-uridine added in trace amounts to the medium. The adenine nucleotide pool was 2.4-fold larger than in situ after 7 hrs of incubation and then returned to values 30% higher than that found in the myocardium after 25 hrs. Cytosine and uracil nucleotide pools after 25 hrs of culture were respectively 2 and 4-fold larger than in situ and remained at these levels thereafter. Intracellular cytidylate and uridylate equilibrated very rapidly with exogenous [U-14C]-cytidine but not with [U-14C]-uridine. We conclude that, under the experimental conditions used here, the synthesis of pyrimidine nucleotides in isolated myocytes is mainly supplied by exogenous nucleosides. Furthermore, extracellular cytidine is rapidly converted to both uracil and cytosine nucleotides while uridine serves only as the precursor for uracil nucleotide synthesis.


Mediatory role of copper in reactive oxygen intermediate-induced cardiac injury. In this report the mediatory role of copper in cardiac injury produced by reactive oxygen intermediates was examined. Isolated rat hearts were perfused with Krebs-Henseleit buffer containing 0.25 mM ascorbate plus varying concentrations of copper-bis-histidial for up to 60 min. Using salicylate as a probe, OH generation by this system was demonstrated. Copper or ascorbate alone had minimal effect on cardiac function as determined by heart rate, coronary flow, left ventricular systolic pressure development, end diastolic pressure and +/- dP/dtmax. Copper, from 0.5 microM to 20 microM, and ascorbate, 0.25 mM, resulted in concentration-dependent decreases in all of the experimental variables. Treatment with 5 or 20 microM copper resulted in complete loss of cardiac function within 40 and 30 min, respectively. By 30 min, 5 microM copper had resulted in increased end diastolic pressure to greater than 40 mmHg. By 60 min, perfusion with 1 microM copper resulted in almost 100% loss of function and end diastolic pressure greater than 25 mmHg. Copper, 0.5 microM, also decreased cardiac function, but to a lesser degree. Catalase, 100 units/ml, was effective in preventing the copper-ascorbate induced cardiac damage while superoxide dismutase, 25 units/ml, was ineffective. Observations by light and electron microscopy demonstrated patchy regions with vacuolization corresponding to swollen mitochondria. These results clearly demonstrate that copper-catalyzed redox reactions can induce cardiac injury via a mechanism which appears to be related to the production of OH.


Cocaine and speedball users: differences in psychopathology. Affective distress and related symptoms associated with co-injected cocaine and opioid ("speedball") use are incompletely explored, and the extent to which they diverge from problems shown by cocaine abusers who do not prefer opioids is unknown. This investigation compared groups of speedball and non-speedball cocaine users on global measures of depression and anxiety and modal groupings of personality characteristics measured by the MMPI. Compared to men who use cocaine without opioids, compulsive speedball users evidenced significantly greater problems with depression, trait anxiety, and related symptomatology, and were more uniformly characterized by modal profiles reflecting severe psychopathology and maladjustment. These results agree with descriptions of severe pathology associated with speedball use.


An incest survivors' therapy group.

An incest survivors' therapy group. A weekly therapy group for women who have been incest victims and are residents of Odyssey House, a therapeutic community for drug abusers, is described. Women participating in the group have a significantly greater retention and graduation rate than other women in the program. The group reduces isolation; helps in dealing with denial; clarifies relations with children, other family members, and significant others; and facilitates dealing with incest-related guilt and shame. The impressive therapeutic progress of the incest survivors, which is attributable to the group, suggests the possible utility of such a group for other therapeutic communities and treatment programs.


Crack cocaine use in a cohort of methadone maintenance patients. We examined crack use in a cohort of methadone patients originally enrolled in 1984-86. Crack use questions were added to the study in 1987. Of the 494 methadone patients originally enrolled, 228 subjects remained in methadone and were re-interviewed in 1987-88, and 234 remained in methadone and were re-interviewed in 1988-89. Approximately one-quarter of the subjects were using crack at each of the 1987-88 and 1988-89 data collection points, and only 3% of the subjects were using crack at daily or greater frequencies at each of the 1987-88 and 1988-89 interviews. Concurrent crack use was associated with (a) the number of noninjected drugs being used; (b) the number of IV drug-using sexual partners; (c) drug injection; and (d) the use of nonheroin opiates. Persistent crack use, defined as use in both 1987-88 and 1988-89, was associated with previous noninjected drug use and previous suicide attempts. While the potential problem of crack use among methadone patients should not be minimized, it appears that, compared to illicit drug injectors not in treatment, being in methadone maintenance may offer a protective effect against crack use.


Improving treatment outcome in pregnant opiate-dependent women. Outcomes for 6 pregnant methadone-maintained opiate-dependent subjects in enhanced treatment were compared to those of 6 women receiving conventional methadone maintenance. Enhanced treatment consisted of weekly prenatal care, relapse prevention groups, thrice weekly urine toxicology screening with positive contingency awards for abstinence, and therapeutic child care during treatment visits in addition to treatment as usual. Treatment as usual included daily methadone, group counseling, and random urine toxicology screening. Study patients differed from the comparison group in three important ways, having fewer urine toxicology screens positive for illicit substances (59% vs. 76%), three times as many prenatal visits (8.8 vs. 2.7), and heavier infants (median birth weight, 2959 vs. 2344 grams). These results suggest that enhanced drug treatment can improve pregnancy outcome and, in particular, reduce low birth weight for this high-risk population.


The treatment of substance abusers diagnosed with obsessive-compulsive disorder: an outcome study. Sixty substance abusers dually diagnosed with obsessive-compulsive disorder (OCD) and voluntarily admitted to a drug-free therapeutic community were randomly assigned to one of three treatment conditions. One group received a combined intervention that addressed their obsessive-compulsive symptoms and substance abuse; a second group received only substance abuse treatment; and an attention control group received treatment for their substance abuse and training in progressive muscle relaxation. Patients who received treatment for their OCD and substance abuse stayed in treatment longer, showed greater reductions in OCD symptom severity, and had higher overall abstinence rates at 12-month follow-up.


The stability of neuropsychological test performance in a group of parenteral drug users. The stability of neuropsychological performance in a sample of drug abusers was investigated for a wide range of neuropsychological tests, using a test-retest paradigm with 16 parenteral drug users. The battery administered included tests of general intellectual function, abstract reasoning, verbal memory, language, attention, visuospatial ability, set switching, speeded performance, and manipulative dexterity. Stability coefficients were of a moderate to high magnitude for most of the tests and were comparable to coefficients found in other studies of non-drug-users. Two exceptions, however, were the Selective Reminding Test and the Perdue Pegboard. Possible reasons for instability with these two tests are discussed. It is concluded that neuropsychological investigations of drug abusers can yield consistent and reliable data, although further studies should employ alternative and/or supplementary measures of verbal memory and motor function.


Controlled use of heroin in patients on methadone maintenance treatment. The efficacy of methadone maintenance treatment was evaluated on 93 patients after 10 years of therapy. On the basis of therapeutic compliance, patients were divided into three groups: (a) 40 Total-Agreement subjects on weekly take-home methadone; (b) 28 Partial-Agreement subjects, who regularly attended the clinic daily but presented episodic positive urinalysis; (c) 25 No-Agreement patients, who were absent from the clinic more than twice a month and had a high rate of urinalysis positive for morphine. Statistical analysis, based on social adjustment improvement and criminality rate decrease, divided the 93 patients into 2 distinct categories. The first category, characterized by high social adjustment and low criminality score, included the Total- and Partial-agreement groups. The second, characterized by significantly lower social adjustment and higher criminality score, included all No-Agreement patients. This suggests that methadone treatment was able to dissociate heroin use from low social functioning. It was concluded that, in a condition of adequate compliance the episodic use of heroin is of no harm to patients on methadone maintenance therapy, that is, methadone maintenance treatment permits a controlled use of heroin.


The Iowa therapeutic community model of chemical dependency treatment. A model of inpatient chemical dependency treatment used in several facilities in Iowa is described. Through combining 12 Step, Therapeutic Community and Reality Therapy concepts, the issues of denial, control, and self centeredness are addressed in developmental phases. Individualizing the program through multidisciplinary input is explained in terms of identifying hindrances in perceiving reality and assisting the patients reduce resistance in a downhill process of increasing wholeness and growth.


Oxygen desaturation during fiberoptic bronchoscopy. A prospective study was performed to detect oxygen saturation (SaO2) during and following fiberoptic bronchoscopy (FOB) in 50 patients. Twenty-five patients (group 1) underwent the procedure without and 25 (group 2) with supplemental oxygen. The SaO2 declined from the baseline value of 96.4% to 92.08% in group 1 and to 94.88% in group 2 after bronchoscopy alone. The decline was also noted when biopsy and broncho-alveolar lavage (BAL) were performed, the lowest values being recorded during BAL. The result showed that the fall in SaO2 in group 2 was significantly less than that in group 1 (P < 0.05). SaO2 returned to baseline values after a mean time of 4.9 minutes in group 1 and 2.4 minutes in group 2, demonstrating the benefit of supplemental oxygen.


Study of arterial aneurysms.

Study of arterial aneurysms. A review of 46 patients operated for arterial aneurysms over an eight years period is presented. Twenty-nine patients developed aneurysm following vascular trauma while 17 had atherosclerotic aneurysms. Thirty-one patients presented with painless swelling or swelling with mild pain and four patients with frank rupture. Reconstruction was successful in all cases with a peri-operative mortality of only 4.3%. Unlike western countries, trauma is the leading cause of aneurysm and aneurysms can be managed with minimal morbidity and mortality.


Coronary risk factors used to predict coronary artery disease by logistic regression analysis. Risk factor analysis in coronary artery disease was conducted in 303 patients who underwent coronary arteriography to identify associations between personal characteristics and the prevalence of coronary heart disease. Age, sex, obesity, smoking, alcohol intake, hypertension, diabetes mellitus, serum uric acid, total cholesterol, LDL- and HDL-cholesterol, triglyceride, and atherogenic indices were statistically analyzed. All 13 variables were first compared between patients with positive and negative ergonovine tests. Only total cholesterol was significantly different, while significant differences in age, sex, history of diabetes, total cholesterol, LDL- and HDL-cholesterol, triglyceride and atherosclerotic indices were observed between patients with and without organic coronary artery stenosis. A multivariate analysis was performed, and the resulting equation was tested using the remaining patients. Logistic analysis of all 13 variables identified 5 (age, sex, diabetes mellitus, LDL- and HDL-cholesterol) which accounted for the differences between patients with and without significant coronary artery disease and that were validated in the test group. The sensitivity for prediction of coronary artery disease was 75.8%, specificity 68.5%, and predictive accuracy 71.5% in the test group. Thus, risk factor analysis appears to be very valuable in screening subjects with high-risk organic coronary stenosis and in optimizing the preventive and therapeutic modalities, but not in predicting vasospastic subjects.


Manganese-containing superoxide dismutase in blood and urine during open-heart surgery. Concentrations of Manganese-containing superoxide dismutase (Mn-SOD) were measured perioperatively by enzyme immunoassay in serial samples of arterial and coronary sinus blood and urine taken from 18 patients undergoing mitral valve surgery. The mean Mn-SOD concentration in the arterial blood samples was 66.2 (SD 16.1 ng/ml) at induction of anesthesia, increased gradually after reperfusion and peaked on the 2nd post-operative day [150 (SD 58.3) ng/ml]. The mean concentration of Mn-SOD in the coronary sinus blood samples was significantly higher than in the arterial samples only at the 6th hour after reperfusion [97 (SD 21.8) ng/ml vs 90.3 (SD 20.9) ng/ml, p < 0.05]. Although concentrations of Mn-SOD in blood did not increase in 8 patients who underwent midline sternotomy for a mediastinal tumor, they increased dramatically in 3 patients who sustained a perioperative myocardial infarction. During open heart surgery the peak values of plasma Mn-SOD concentrations were correlated to that of plasma creatine kinase-MB concentrations (r = 0.5532, n = 18, p < 05) and cardiac ischemic period (r = 0.5186, n = 18, p < 05). Although the meaning of an increase in plasma Mn-SOD concentrations during open heart surgery is not clarified, it may be released from the heart and anywhere also in the body damaged during cardiopulmonary bypass.


Changes in serum lipoprotein(a) and C4b-binding protein levels after acute myocardial infarction. We investigated changes in serum lipoprotein(a) (Lp(a)) and C4b-binding protein (C4bp) levels following acute myocardial infarction (AMI). Serum lipids, apolipoproteins, C-reactive protein (CRP), sialic acid and haptoglobin (Hp) were also measured and evaluated. The study involved 16 patients (11 men, 5 women, mean age 68.2 +/- 8.4 years) with AMI admitted within 12 h after the onset of illness. CRP rose sharply and immediately after the onset of AMI, reaching its maximum level on the 3rd day of illness. This was followed by temporary increases in sialic acid and Hp, both of which peaked on the 7th day of illness. The Apo A-I level was significantly lower on the 14th day, while the Apo B level was significantly lower on the 3rd day. Serum total cholesterol (T-Ch), HDL-Ch and LDL-Ch showed temporary decreases after the onset of illness. Serial examination of the serum of patients with AMI yielded the following findings; 1) Both Lp(a) and C4bp levels rose temporarily with similar patterns of variation, and 2) they took longer time to peak (around the 14th day) and lasted longer than the increases in the other acute reactive proteins.


Nine-lead Holter monitoring using a lead-switching technique for the detection of ischemic ST changes. We devised a technique to record a greater number of leads (a 9-lead monitor) by connecting a lead-switching adaptor to a commercially available 3-channel Holter recorder (9-lead DCG). Anodes were attached from positions V1 to V6 (CM1 to CM6), and to high lateral (HL), low lateral (LL) and low back (LB). A cathode was attached to the manubrium of the sternum. The CM5 lead was continuously recorded on channel 1. The device is able to switch continuously every 20 sec among leads CM6, LB, HL and LL on channel 2, and among leads CM1, CM4, CM2, and CM3 on channel 3. Electrocardiograms were simultaneously recorded with both the 9-lead DCG and the conventional 12-lead ECG systems during treadmill testing in 67 patients with coronary artery disease. In addition, 6 patients with acute myocardial infarction were studied with the 9-DCG to test ST elevation. The sensitivity and specificity of leads CM2 to CM6, HL, LL, and LB in detecting ST depressions that occurred in each corresponding lead of the 12-lead ECG lead were very high (p < 0.0001). The LB lead in particular was noteworthy for its markedly high specificity (94%) and sensitivity (83%) in detecting ST depressions occurring in leads II and aVF. Our lead-switching technique is useful when an increase in the number of leads is required in the DCG method. The LB lead, an anode on the low back, may be specific to detect inferior myocardial ischemia.


HA-1077 suppress both proliferation of vascular smooth muscle cells and c-fos mRNA induction. HA1077 is a newly synthesized vasodilator with unique intracellular calcium antagonistic action. In this study, its effect on the growth of vascular smooth muscle cells (VSMC) stimulated by fetal calf serum was examined. Both the proliferation and [3H]thymidine incorporation into DNA of the growth-arrested VSMC was dose-dependently inhibited by HA1077. The expression of a proto-oncogene, c-fos, which reached the maximum 30 min after addition of serum, was similarly inhibited by this agent in a dose-dependent manner. Thus, HA1077 is expected to be a useful vasodilator agent capable of suppressing the growth of VSMC which is thought to be an important underlying mechanism of atherosclerosis or restenosis after angioplasty.


Effect of a prostacyclin derivative (OP-41483) and a hyperosmotic agent (glycerol) on brain edema and metabolism in cerebral ischemia. A PGI2 derivative, OP-41483, and a hyperosmotic agent, glycerol, were tested for possible beneficial effects on brain edema, metabolism and pathological changes in cerebral ischemia. Combination treatment with these agents was also tested. Cerebral ischemia was produced in spontaneously hypertensive rats, using bilateral common carotid artery ligation (BLCL). OP-41483 was administered four times, hourly (500 ng/kg x 4, i.p.). Ten percent glycerol was administered intravenously (6.6 ml/kg). And, for the combination treatment, OP-41483 was administered three times, hourly (500 ng/kg x 3, i.p.), and 10% glycerol was administered intravenously (6.6 ml/kg) in the same manner as the glycerol treated group. In ischemic controls, saline was administered intravenously (6.6 ml/kg). After 3 h of ischemia, brain water content and metabolites were determined and pathological observation was conducted using electron microscopy. OP-41483 treated animals maintained higher levels of ATP concentration and reduced accumulation of lactate, but showed no difference in brain water content compared to saline treated controls. Glycerol treated animals showed significance in terms of reduction of brain water content and accumulation of lactate. Glycerol abated the depletion of ATP concentration. OP-41483+glycerol treated animals showed the most significant effect on the reduction of brain water content and accumulation of lactate. The combination treatment also maintained higher levels of ATP concentration. Additionally, swelling of astrocytic foot processes and mitochondria with destroyed crista were not observed pathologically in the combination treated animals. These results show that OP-41483, glycerol and combination treatment are beneficial in the treatment of cerebral ischemia. They also indicate that the combination treatment significantly enhances the protective effects compared to individual treatment.


Two cases of variant form angina pectoris associated with myocardial bridge--a possible relationship among coronary vasospasm, atherosclerosis and myocardial bridge. Myocardial bridge (MB) is a congenital anomaly of the coronary artery and may occur in 5 to 12% of the human population. However, the mechanism of MB-induced myocardial ischemia is still speculative. We report 2 cases of variant form angina pectoris associated with MB in which myocardial ischemia seemed to be related to the interaction between coronary perfusion and MB. In case 1, electrocardiography during anginal attack at rest showed ST elevation in the inferior leads and MB was observed after percutaneous transluminal coronary angioplasty at the site of the right coronary artery lesion following successful dilatation. In case 2, MB of the left anterior descending coronary was located in the identical portion where coronary vasospasm was induced by intracoronary acetylcholine injection, although ischemia during the spontaneous anginal attack was limited to the inferior area of myocardium. These 2 cases suggest that MB can be, at least in some patients, one of the possible causes of the endothelial damage which seems to be related to coronary vasospasm; this was documented in both cases.


Adult respiratory distress syndrome in a patient with uncomplicated acute myocardial infarction. A 53-year-old woman was admitted with a diagnosis of acute extensive anterior myocardial infarction. Pulmonary capillary wedge pressure was 17 mmHg, and cardiac index 2.4 l/min.m2. The patient was successfully treated with diuretics and nitrates but on the fifth hospital day moist rales were noted over the entire lung field. A chest roentgenogram showed diffuse bilateral pulmonary infiltration with unchanged cardiac silhouette. Arterial oxygen partial pressure was reduced to 45 mmHg under 3 liters of nasal oxygen inhalation. Pulmonary capillary wedge pressure was 8 mmHg. Diffuse infiltration was not ameliorated by intravenous injection of diuretics. The diagnosis of adult respiratory distress syndrome was finally established by successful treatment with steroids.


[Analgesic effects of epidurally administered fentanyl for postoperative pain relief--comparison with buprenorphine]. We did a retrospective study on 177 patients after upper and lower abdominal surgery, and compared the efficacy of epidural administration of fentanyl and that of buprenorphine for postoperative pain relief. In fentanyl (F) group, 73 patients received fentanyl 0.1 mg with saline 8 ml epidurally after operation, followed by a constant rate infusion of 0.025 mg.hr-1 for 18-24 hrs. In buprenorphine (B) group, 104 patients, received buprenorphine 0.2 mg with saline 9 ml epidurally. After upper abdominal surgery, 33 patients (76.7%) in F group and 27 patients (52.9%) in B group obtained satisfactory analgesia (P < 0.05). The difference of the degree of analgesia after lower abdominal surgery was not significantly different in both groups. Respiratory depression occurred in 19 patients in B group and 5 patients in F group (P < 0.05). It is concluded that epidural fentanyl delivered by continuous infusion offers a significant advantage compared with epidural buprenorphine for postoperative pain relief following upper abdominal surgery.


[Continuous epidural infusion of bupivacaine and buprenorphine for postoperative pain relief]. The efficacy for postoperative analgesia and side-effect of combined epidural infusion of bupivacaine and buprenorphine in comparison with each of these drugs alone were evaluated in 150 patients. All patients received initially bupivacaine 8 ml and buprenorphine 0.1 mg. In a random order, epidural infusion of 5 micrograms.ml-1 buprenorphine 1 ml.h-1 (group A; n = 50), 0.25% bupivacaine 1 ml.h-1 (group B; n = 50), or a combination of the two drugs 1 ml.h-1 (group C; n = 50) was continued for 48 h by a portable disposable device. The analgesic efficacy in group C was superior to that in group A or group B. No significant difference in the incidence of side-effect was found among the three groups. We conclude that epidural analgesia with the combination of buprenorphine and bupivacaine is safe, and easy to manage, giving pain relief superior to that provided by each of these drugs alone.


[The antinociceptive effects of fentanyl, midazolam and clonidine and their interactions in the spinal dorsal horn]. Lamina V-type neurons on the spinal dorsal horn which responded to the bradykinin injection into the femoral artery were studied neurophysiologically in the spinal transected cats by the tungsten microelectrode method. It has been demonstrated that the separate and combined antinociceptive effects of fentanyl, clonidine and midazolam administered intrathecally can produce reduction in response to noxious stimuli. Fentanyl (25 micrograms), clonidine (30 micrograms) and midazolam (1.0 mg) separately suppressed noxious evoked activity at the spinal level. On the other hand, fentanyl (5 micrograms), clonidine (5 micrograms) and midazolam (0.5 mg) each produced no significant suppression of the evoked activity. However, the combinations of drugs at lower doses produced supra-additive suppressive effect. These suppressive effects were reversed by each antagonist (naloxone, yohimbine and flumazenil). These findings suggest that when two of these drugs are combined at subanalgesic doses, a significant synergistic interaction is exerted. Therefore, the use of these drugs in combination can reduce the total amount of any one drug required for analgesia in the spinal cord and also reduce the side effects of these agents.


[Hemodynamics and serum nitroglycerin concentration during intraoperative use of nitroglycerin patches]. We investigated the effect of number of nitroglycerin (TNG) as patches, containing 5 mg of TNG per sheet, on hemodynamics in 29 adult patients undergoing elective surgery. Patients were randomly allocated to four groups according to the number of the sheets applied (0, 2, 5 and 10 sheets). Hemodynamics as well as plasma TNG concentration were measured at 4 points (just before and 30, 60 or 120 minutes after application of TNG patch) in each group (n > or = 6). Plasma TNG concentrations depended on the number of the sheets and increased until 120 min after the application. Mean arterial pressure decreased gradually and significantly after applying two sheets or more. However, no relation was observed between the plasma concentration of TNG and the decrease in mean arterial pressure. We conclude that induced hypotension can not be intensified by increasing the number of the sheets of TNG patch.


[Change in arterial ketone body ratio during gastrectomy and mastectomy]. We evaluated the influence of operative procedure on arterial ketone body ratio (AKBR), which indicates the function of the liver cells, in patients undergoing gastrectomy or mastectomy. AKBR during the stomach resection was significantly lower than that during the breast resection. A significant reduction in AKBR due to induced hypotension was observed in mastectomy group. SGOT and SGPT increased significantly in gastrectomy group, but unchanged in mastectomy group on the first and the seventh postoperative days. On the first post-operative day, SGOT and SGPT in gastrectomy group were significantly higher than those in mastectomy group. These findings suggest that influence of operative procedure on the liver during gastrectomy is more serious than that during mastectomy.


[Perioperative blood glucose levels in children]. The perioperative blood glucose (BG) level may represent autonomic responses to various stresses by anesthesia and surgery. We studied the effects of regional blocks combined with general anesthesia on the BG response in pediatric patients. We could not find any significant differences in BG levels between blocked and non-blocked patients. However, BG levels of the children who had been hysterical or crying during the induction of anesthesia were significantly higher than those of the children who had been calm or sleeping. Forty healthy children, aged 1-3 yrs who were scheduled for elective urological surgery were studied. Anesthesia was induced with halothane in oxygen. The anesthesiologist recorded the children's attitude before and during the induction of anesthesia, and described as "calm or asleep (Calm-group), and hysterical or crying (Crying-group)". Patients were assigned randomly to two groups as follow; Group A (n = 20): receiving nerve blocks, Group B (n = 20): without the blocks. The anesthesia was maintained with 1-2.5% halothane in oxygen required to maintain the hemodynamic parameters within 10% of baseline value before surgery. Both groups received lactated-Ringer's solution during the study period. The venous BG levels were determined 5 times, i.e. immediately after asleep, 5, 15, 30 minutes after skin incision and at the conclusion of surgery. BG levels were compared between groups using Mann-Whitney Test. There was no significant difference in BG levels between Group A and B throughout the study period. But BG values of Crying-group (n = 9) were significantly higher than those of Calm-group (n = 31) during perioperative period.(ABSTRACT TRUNCATED AT 250 WORDS)


[The effect of ionized calcium concentration on muscle contraction and on the effect of non-depolarizing neuromuscular blocking agent]. The influence of serum ionized calcium (Ca2+) on the muscle contraction and on the effect of non-depolarizing neuromuscular blocking agent, d-tubocurarine (dTc), was investigated. The isolated phrenic nerve-diaphragma preparation of the rat was stimulated directly and indirectly at 0, 0.4, 0.7, 1.4, 2.5 mM of total calcium concentration (Ca2+: 0, 0.39, 0.52, 1.12, 2.32 mM, respectively). With indirect stimulation, no contraction was observed from 0 to 0.4 mM of calcium, but the twitch tension returned to 100 percent of control at over 0.7 mM of calcium. However, with direct stimulation, the twitch tension was obtained up to 100 percent of control independent of Ca2+ concentration. The ED50 of dTc was correlated well with the serum Ca2+. These findings suggest that the decrease of serum Ca2+ concentration interferes with the muscle contraction and potentiates the effect of dTc. We speculate that the site of its action may be primarily prejunctional.


[Effects of magnesium and calcium on muscle contractility and neuromuscular blockade produced by muscle relaxants and aminoglycoside]. The muscle contractility and neuromuscular blockade of muscle relaxants are influenced by the electrolytes, especially magnesium ion(Mg2+) and calcium ion(Ca2+), in the extracellular fluid. The present study was designed to evaluate the effects of Mg2+ and Ca2+ on muscle contractility and on the blocking properties of d-tubocurarine, succinylcholine and aminoglycoside antibiotics, tobramycin in vitro, using rats' phrenic nerve-hemidiaphragm preparations. Mg2+ inhibited the twitch tensions in a concentration dependent manner. A low concentration of Ca2+ decreased the twitch tensions, but a high concentration of Ca2+ had no effect on them. Mg2+ potentiated the block produced by d-tubocurarine, succinylcholine and tobramycin depending on the concentration of Mg2+, and good regression lines were obtained. Ca2+, however, antagonized the neuromuscular blockade of the three drugs at any level of Mg2+ concentration. From these findings, it should be noticed that special care must be taken to prevent a profound neuromuscular blockade due to the drug interaction of each drug when a muscle relaxant or an aminoglycoside antibiotics is administered to a patient suffering from hypermagnesemia and/or hypocalcemia.


[Effect of atropine and pirenzepine on bronchoconstriction during vagus nerve stimulation in dogs]. The effects of atropine or pirenzepine on bronchoconstriction and reduction in heart rate during vagus nerve stimulation were studied in 10 anesthetized dogs. The bronchoconstriction produced by electrical stimulation of the vagus nerves, 5 min after intravenous injection of atropine or pirenzepine, was recorded as an increase in pulmonary inflation pressure. Atropine and pirenzepine at the doses of 0.01 mg.kg-1 and 0.2 mg.kg-1 respectively were administered and supplemented twice at 30 min intervals. Atropine inhibited the reduction in heart rate more than pirenzepine at all doses. Pirenzepine inhibited the increase in pulmonary inflation pressure more than atropine. Our results suggest that pirenzepine which inhibits bronchoconstriction is useful in patients with bronchial asthma and/or COPD as an anticholinergic agent.


[Quantitative analysis of electroencephalographic (EEG) activity during sevoflurane anesthesia]. The change in electroencephalographic (EEG) activity during sevoflurane anesthesia was quantitatively evaluated by using zero-crossing analysis in 10 adult patients. The deceleration of EEG activity was significant and dose-dependent. Such change was not significantly different among the regions. Burst suppression appeared at 2-2.5 MAC in all patients. The significant decrease in both alpha and beta activities and the significant increase in delta activity disappeared with 1% or less of sevoflurane. Toward the emergence from anesthesia, EEG activity accelerated and there was no significant difference in its activity between the emergence period and control period. The results demonstrated that the effect of sevoflurane on EEG activity is similar to that of other inhalation anesthetics, but the difference among anesthetics should be clarified in a further study.


[Anesthetic induction of children with high concentrations of sevoflurane]. The induction and intubation with high concentrations of sevoflurane (4% or 6.4%) were investigated in 180 children with ages ranging from 1 to 6yr 12m. Average induction time (duration between the start of induction and loss of consciousness) with 4% sevoflurane was 56 sec and that with 6.4% was 47 sec. Eight patients out of 90, to whom 4% sevoflurane was applied, could not be intubated due to closure of vocal cord, cough or closure of mouth. The most serious side effect was convulsion during induction, of which incidence was 6% of all patients, and occurred more frequently with 6.4% sevoflurane. The convulsion spread from hand on one side to all over the body, and disappeared spontaneously within 30 sec. There was no significant side effect in cardiovascular system, such as hypotension, arrhythmia or bradycardia. In conclusion, smooth intubation was possible without use of muscle relaxants with high concentrations of sevoflurane. This can be one of the choices for induction of children, though there is a possibility of occurrence of convulsion.


[Amrinone reverses cardiac depression by enflurane in the dog]. To evaluate the interaction of amrinone with inhalational anesthetics, cardiovascular effects of amrinone were investigated in nine mongrel dogs anesthetized with enflurane. Each dog received enflurane and amrinone in the following sequence: 1) 2% enflurane alone, 2) continuous infusion of 20 micrograms.kg-1.min-1 during enflurane, 3) 40 micrograms.kg-1.min-1 infusion during enflurane. Amrinone 40 micrograms.kg-1.min-1 during enflurane anesthesia improved the maximum left ventricular dP/dt, stroke volume and decreased effective arterial elastance (Ea) without changes in left ventricular end-diastolic pressure and heart rate. Left ventricular pressure (LVP) and systolic femoral arterial pressure were stable, but diastolic femoral arterial pressure decreased significantly from enflurane anesthesia alone. These parameters at 20 micrograms.kg-1.min-1 of amrinone infusion during enflurane showed the same tendency with 40 micrograms.kg-1.min-1 infusion but not significantly different from enflurane alone. This result suggests that amrinone may be beneficial in the patients with depression of cardiac performance during anesthesia.


[Appropriate puncture site for CT monitored celiac plexus block determined from CT films]. We investigated appropriate puncture site, angle of needle entry, and the distance of the insertion for CT monitored celiac plexus block using CT photograms on prone position in sixteen patients with gastrointestinal diseases. In retrocrural approach during CT monitored celiac plexus block, the average distances of puncture sites to midline were 3.84 cm on the right and 4.06 cm on the left, and the average needle angles were 74.1 degrees on the right and 76.9 degrees on the left. The average depths from the puncture site to marked position in retrocrural space were 7.88 cm on the right and 7.85 cm on the left. The rates of predicted organ puncture when needle is inserted 7 cm lateral to the midline of the spinal process were 43.8 percent in the right lung, 12.5 percent in the left lung, 56.2 percent in the right kidney and 68.8 percent in the left kidney. From these results, we found high possibilities of organ injuries using the conventional technique for celiac plexus block. We conclude that we could perform celiac plexus block more safely and surely using the retrocrural approach by CT monitoring, as serious complications are avoidable by viewing ideal puncture course on CT photographs.


[A role of statistical thinking in clinical anesthesia practice]. The world where we practice anesthesia is composed of heterogeneic, multivariate components. Yet, these components are not known with certainty. Nevertheless, we are compelled to make a prediction and, furthermore, to make a decision to treat patients at risk. Therefore, decision making under uncertainty is one of the important issues in clinical practice. A fundamental strategy for such action would be one of statistical thinking process, which is widely applicable in many circumstances to reduce the degree of uncertainty and to construct a tool to deal with uncertainty. Here, topics relevant to statistical thinking and its pitfalls are discussed, including those related to information theory. In short, statistical thinking will, I believe, remain to be valid to make an appropriate prediction and decision in the years to come.


[Anesthesia for a patient with pemphigus erythematosus]. An 82-year-old female with pemphigus erythematosus had the head of femur replaced. In her case, systemic lupus erythematosus and myasthenia gravis, typical complications of pemphigus erythematosus, were not present. As even slight friction easily causes blisters in patients with pemphigus, we paid attention mainly to fragility of the skin and the mucus membranes and were careful to minimize physical stimulation of the skin. Because the mucosa of the pharynx or oral cavity could easily be damaged and form blisters during intubation, which in turn might make the management of the upper airway more difficult, we selected spinal anesthesia. No serious perioperative complications, except a few blisters around the wound were observed. We thus successfully managed anesthesia of a patient with pemphigus erythematosa.


[Deep neck infection following endotracheal intubation]. A 71-year-old woman was transferred to Kushiro City General Hospital because of fever, sore throat, diffuse neck swelling and dyspnea. She had received right mastectomy for breast cancer under general anesthesia 6 days before the admission. The lateral X-ray film of the neck revealed abscess in the retropharyngeal space and the retroesophageal space. CT scan revealed mediastinitis. Next day she received neck dissection for drainage of the abscess under general anesthesia. Although the posterior pharyngeal wall was swollen, endo-tracheal intubation was not difficult. Brown tinged and purplish pus was aspirated from the interspace of carotid sheath and trachea, the retropharyngeal space, and the superior mediastinal space. The infected site was irrigated with a lot of peroxide and saline and draining tubes were placed in each interspace. Tracheostomy was not done but the patient was admitted to the ICU with her trachea intubated. The day after operation, she was extubated. Three days after the operation chest X-ray revealed pyothorax and chest tube was inserted for drainage. Seven days after the operation she was transferred to the ENT ward. Thereafter her recovery course was uneventful. It seems that the deep neck infection was probably caused by the injury on endotracheal intubation at the first operation in this case. Although this patient was cured of mediastinitis following deep neck infection, which is still lethal, early diagnosis and surgical drainage of the abscess are necessary.


[Coronary artery spasm immediately after the long-standing operation for cancer of the tongue]. A 72-year-old male underwent radical operation for cancer of the tongue. Anesthesia was maintained with the combination of enflurane-N2O-vecuronium and cervical epidural block. Five minutes after the cessation of the longstanding operation, VT and circulatory collapse occurred. After administration of lidocaine and ephedrine, VPC and ST elevation were noted, followed by VT and Vf. Cardioversion successfully restored sinus rhythm with no ST change, suggesting an episode of coronary artery spasm. The possible inducing factors in this case were hypotension and acute imbalance in autonomic nervous systems caused by hypovolemia, hypothermia, insufficient anesthetic depth, loss of surgical stress, neostigmine and epidural block. The authors reviewed case reports on coronary spasm, especially looking for possible inducing factors of coronary artery spasm during anesthesia.


[A case report of a burn produced by the probe of a pulse oximeter]. We experienced a case of burn produced by the pulse oximeter probe at the finger of a patient. Patient was a 75-year old man scheduled to transurethral prostatectomy. The pulse oximeter probe made by Physio-Control Co. was attached at his finger and was connected to a pulse oximeter manufactured by Ohmeda Co. Soon after the start of Spo2 measurement the anesthetist became aware of abnormal heating of the probe and removed it immediately. But unfortunately a small thermal injury was formed at patient's finger. Abnormally excessive current went through the detector diode and heated the finger probe. Attention to correct connection between the pulse oximeter probe and the finger is necessary before use.


[An unusual cause of anesthetic ventilator malfunction]. We report a case of malfunction of an anesthetic ventilator by an unusual cause. A 48-year-old male with gastric cancer was scheduled for gastrectomy. Anesthesia was maintained with enflurane, N2O, O2 and epidural blockade using a semiclosed circuit system. The patient was ventilated using AV1 anesthetic machine (Dräger Co.). Forty minutes after induction of anesthesia, chest movement of the patient suddenly stopped. There was no gas flow to the patient during inspiratory phase. Air leak was not found in anesthetic respiratory circuit and at the bellows of the ventilator. The supply of oxygen and air to the anesthetic machine was sufficient. Since we could not find any cause of the ventilator failure, anesthesia was maintained with manual ventilation by using another anesthetic machine until completion of the surgery. After the surgery, we recognized that the controller unit of expiratory valve of the ventilator was obstructed by a Tamper Proof Film, which seals the outlet of a commercial bag of lactated Ringer's solution (Solulact, Terumo Co.). It seems that the film dropped accidentally between the main part and the ventilator system of anesthetic machine when the bellows was exchanged before the surgery, and moved on to the controller unit of the expiratory valve of the ventilatory system during surgery. In conclusion, it is necessary for anesthetists to understand the inner structure and system of the anesthetic machine and to check the anesthetic machine to avoid the troubles and accidents related to anesthetic machine.


[Sevoflurane anesthesia for asthmatic patients: report of five cases]. We anesthetized five patients receiving antiasthmatics with sevoflurane-nitrous oxide-oxygen. Induction with sevoflurane inhalation was conducted uneventfully in all patients. Mild asthmatic attacks occurred in three patients during light anesthesia. The asthmatic attacks subsided with elevating the concentration of sevoflurane or the administration of aminophylline in two patients. In only one patient, inhalation therapy with salbutamol was required to treat an asthmatic attack during deep anesthesia. The results indicate that sevoflurane was a proper inhalation anesthetic for our asthmatic patients.


[The effect of stellate ganglion block therapy on tonsillitis]. We examined a 35-year-old male patient with recurrent tonsillitis who did not respond to operation and drug therapy and we treated him by stellate ganglion block (SGB) alone. The clinical symptoms improved by the SGB. There was no subsequent recurrence. The result suggests that SGB therapy is effective for recurrent tonsillitis.


[A 5-year-old patient with allergic diseases responsive to stellate ganglion block]. We examined a 5-year-old boy with allergic rhinitis, chronic sinusitis, atopic dermatitis, and tendency to suffer common cold, who responded to stellate ganglion block (SGB). SGB therapy was extremely effective for this patient. The result suggests that SGB therapy should be performed on patients with allergic diseases resistant to drug and diet therapy. Furthermore, the indication for nerve block therapy, such as SGB, may be extended even to pediatric patients.


[Evaluation of in vitro antimicrobial activity of cefazolin alone and in combination with cefmetazole or flomoxef using agar dilution method and disk diffusion method]. Antimicrobial activities of cefazolin (CEZ) against 251 strains of various clinical isolates obtained during 1989 and 1990 were determined using the Mueller-Hinton agar dilution method at an inoculum level 10(6) CFU/ml. The reliability of the disk susceptility test was also studied using Mueller-Hinton agar and various disks at inoculum levels of 10(3-4) CFU/cm2 in estimating approximate values of MICs. In addition, antimicrobial activities of CEZ and cefmetazole (CMZ) or flomoxef (FMOX) in combination were investigated against methicillin-sensitive and -resistant Staphylococcus aureus (MSSA and MRSA) using the checkerboard agar dilution MIC method and the disk diffusion test either with the disks contained CEZ, CMZ, and FMOX alone, or CEZ, and CMZ or FMOX in combination. In this study, the MICs of CEZ against S. aureus were distributed with the 3 peak values at 0.39 microgram/ml, 3.13 micrograms/ml and > 100 micrograms/ml. MICs against MSSA were 0.39 microgram/ml to 0.78 microgram/ml, whereas those against MRSA were greater than 0.78 microgram/ml. MICs against majority of strains of Enterococcus faecalis were 25 micrograms/ml. Over 90% of strains of Escherichia coli and Klebsiella pneumoniae were inhibited at the level of 3.13 micrograms/ml. About 60% of isolates of indole negative Proteus spp. were inhibited at the levels of less than 3.13 micrograms/ml and 100% at 6.25 micrograms/ml, but MICs against indole positive Proteus spp., Serratia spp. and Pseudomonas aeruginosa were over 100 micrograms/ml. The antimicrobial activities of CEZ against these clinical isolates were not significantly different compared to those reported about 15-20 years ago, except for S. aureus. Highly resistant strains of S. aureus to CEZ were more prevalent in this study. The inhibitory zones obtained with the disk test were compared with MICs. The results of CEZ disk susceptibility test with 30 micrograms disk (Showa) or 10 micrograms disk (prepared in this laboratory) were well correlated with MICs (r = -0.837 and -0.814, respectively), showing the reliavility of the disk method in estimating approximate values of MICs. In the 4 category classification system currently used in Japan, break points in MIC values proposed are () MIC < or = 3 micrograms/ml, (++) > 3-15 micrograms/ml, (+) > 15-60 micrograms/ml, (-) > 60 micrograms/ml. The results obtained with 30 micrograms disks showed false positive in 7.7% and false negative in 6.8% of the samples. The disk results with E. faecalis showed a higher ratio of false positive results.(ABSTRACT TRUNCATED AT 400 WORDS)


[Antimicrobial activity of AO-128, a novel inhibitor of alpha-D-glucosidase, against anaerobic bacteria]. The in vitro antimicrobial activity of AO-128, an inhibitor of alpha-D-glucosidase, was evaluated against anaerobic bacteria of 45 reference strains (12 genera, 44 species). AO-128 inhibited no strains tested at a concentration of 1,600 micrograms/ml. The results strongly suggested that this compound would not have any influence on the human intestinal microflora, a majority of which is composed of anaerobic bacteria.


[The penetration of cefpiramide into the skin]. To investigate the skin penetration of cefpiramide (CPM, SM-1652), a broad-spectrum and long acting cephem antibiotic, 1 g of CPM was administered by single bolus intravenous injection to patients under general anesthesia during operations for full-thickness skin grafting. The CPM levels in both the serum and the skin were determined by bioassay at specified time intervals, and the following results were obtained. 1. Peak CPM concentrations in the serum (mean: 292.78 micrograms/ml) were observed 10 minutes after administration, and declined very slowly thereafter. 2. Peak CPM concentrations in the skin (mean: 23.05 micrograms/g) were observed 2 hours after administration, and then also declined very slowly. 3. The ratio of skin to serum concentrations was 16% when the peak CPM concentrations in the skin were obtained (2 hours after the administration). These results show that effective CPM concentrations were maintained in the skin for a long period, indicating a good therapeutic action against skin infections.


[Ceftazidime in the treatment of meningitis caused by multiresistant Serratia marcescens]. A patient of subarachnoid hemorrhage was treated with spinal CSF drainage. Serratia marcescens meningitis occurred because of the spinal CSF drainage. The organism was multiresistant and refractory to antibiotics including piperacillin, imipenem, gentamicin and cephaloridine. It was sensitive to ceftazidime (CAZ). Treatment with CAZ resulted in clinical improvement associated with rapid clearing of the organism from CSF. CAZ serum level was high enough and CAZ penetration into the CSF was satisfactory. According to the evaluation of CAZ concentrations in serum and CSF, two regimens of treatment were recommended. One is an administration of CAZ 1 g x 4 times/day. Another is a combination with CAZ administration 2 g x 2 times/day and followed by 1 g x 4 times/day. The results suggest that CAZ is an extremely effective antibiotic for meningitis caused by CAZ-susceptible bacteria.


[A clinical study of combined therapy of aspoxicillin and ceftazidime on intractable respiratory infections]. Both aspoxicillin (ASPC) and ceftazidime (CAZ) were administered together to 55 patients with intractable respiratory tract infections. ASPC and CAZ were administered at daily doses of 4 g and 2 to 4 g, respectively. Clinical effects were excellent in 11 cases, good in 33, fair in 7 and poor in 4, thus the efficacy rate was 80.0%. Bacteriologically, identified organisms were eradicated in 14 cases out of 21, decreased in 3, exchanged in 2 and unchanged in 2. The eradication rate was 76.2%. As a side effect, diarrhea was found in only one case, and abnormal laboratory test values were observed in 4 cases. However, these adverse reactions were not severe, therefore it was possible to continue the chemotherapy. These results indicate that the combination antimicrobial chemotherapy of ASPC and CAZ is effective against intractable respiratory tract infections.


[Clinical effects of cefodizime in severe infections complicated with hematological diseases. Hanshin Infection Study Group]. One hundred thirty-eight patients with severe infections accompanying hematological diseases were treated with cefodizime (CDZM). CDZM was administered by intravenous injection at daily doses ranging from 2 g to 8 g for about 3 to 18 days. Clinical efficacies of CDZM were excellent in 30 cases, good in 37 cases, fair in 1 case and poor in 53 cases. The efficacy rate was 55.4%. The efficacy rate was 56.4% in patients from whom causative organisms were not identified. The efficacy rate against Gram-negative bacteria (66.7%) was higher than that against Gram-positive bacteria (28.6%). No significant differences in the efficacy rates were found among groups of patients with different initial neutrophil counts (less than 100, 100-499, and over 500/microliters). Skin eruption caused by CDZM was observed in 2 patients (1.5%). Hepatic disorders, renal dysfunction and urinary test abnormality (protein and urobilinogen) were observed in 7 (5.0%), 1 (0.7%) and 1 (0.7%), respectively. Therefore, CDZM was considered to be a clinically useful antibiotic for severe infections complicated with hematological diseases.


[Combined effects of arbekacin with other antibiotics against methicillin-resistant Staphylococcus aureus. III. Combined effects of arbekacin with cefotiam or cefuzonam]. Antibacterial effects of combination use of arbekacin (ABK) with cefotiam (CTM) or cefuzonam (CZON) were evaluated against methicillin-resistant Staphylococcus aureus (MRSA) and the following results were obtained. 1. Antibacterial effects of combinations of ABK with CTM and with CZON were equally potent against MRSA at clinically expected 1 MIC of ABK in blood. However, at a sub MIC of ABK the different effects were observed between the 2 combinations. The antibacterial effect of the former was strong and that of the latter was a little weak. 2. In either combination the potency of the antibacterial activity was less dependent on the concentration of CTM or CZON, but was strongly dependent on ABK concentrations. These results suggest that antibacterial effects of the combinations were highly dependent on antibacterial potency and concentration of ABK as previously reported for combinations of ABK with other drugs. 3. The combination use of ABK with CTM appears to be useful in cases of infection by MRSA alone while the combination use of ABK with CZON appears to be useful in cases of double infection with MRSA and Gram-negative bacterium.


[Combined effects of arbekacin with other antibiotics against methicillin-resistant Staphylococcus aureus. IV. Combined effects of arbekacin with cefmetazole or flomoxef]. Antibacterial effects of combination use of arbekacin (ABK) with cefmetazole (CMZ) or flomoxef (FMOX) were evaluated against methicillin-resistant Staphylococcus aureus (MRSA) and the following results were obtained. 1. Antibacterial effects of combinations of ABK with CMZ and with FMOX were equally potent against MRSA at clinically expected 1 MIC of ABK in blood. However, at a sub MIC of ABK different effects were observed between the 2 combinations. The former combination was slightly less effective than the latter. 2. In either combination the potency of the antibacterial activity was less dependent on the concentration of CMZ or FMOX, but was strongly dependent on ABK concentrations. These results suggest that antibacterial effects of the combinations were highly dependent on antibacterial potency and concentration of ABK as previously reported for combinations of ABK with other drugs. 3. It appears that the antibacterial activity of the combination of the sub MIC of ABK with a beta-lactam is an important point in considering the effectiveness of a combination therapy.


[Experimental study on intramaxillary injection of fosfomycin. Effects of fosfomycin on experimental acute sinusitis in rabbits]. We studied effects of intramaxillary injection of fosfomycin (FOM) on experimental sinusitis in rabbits. The experimental sinusitis was induced by intramaxillary injection of Staphylococcus aureus to rabbits for 3 successive days. 1. 0.5, 1, 3 or 5% FOM with saline as a control was instilled into the maxillary sinus and the maxillary sinus mucosa were examined macroscopically and light and electron microscopically. 3% and 5% FOM suppressed the damage of mucosa macroscopically and scanning electron microscopically. 2. After administration of 3% FOM and saline twice a week, the maxillary sinus mucosa was examined macroscopically and light and electron microscopically. The maxillary sinus injected with 3% FOM showed almost normal mucosa after 2 weeks while that injected with saline showed severe mucosal damage. S. aureus were decreased by 3% FOM injection and not found in the maxillary sinus in a week. The results indicate that intramaxillary injection of FOM is very effective in the treatment of sinusitis.