Long-term observation of plasma and tissue enzyme activities in the rat.

During a long-term study in the rat some enzyme activities were determined in plasma, lung, spleen and skeletal muscle. Twelve rats of each sex were investigated every 49 days from 35 until 1115 days of life. Lactate dehydrogenase in lung and spleen decreases; in muscle and plasma, however, the activity varies considerably. Malate dehydrogenase in the tissues remains nearly unchanged apart from distinct peaks in the first year of life; in plasma the activity takes an M-shaped course. In contrast to the changes of glutamate dehydrogenase in the tissues with a tendency to diminish, this enzyme increases in plasma during the lifetime. Aspartate aminotransferase activity in the tissues, except muscle, varies with a rhythmical behaviour, and in plasma shows a gradual increase. Alanine aminotransferase in lung and spleen has two activity peaks. In muscle this enzyme varies only slightly after a steep initial decrease. In plasma the activity has a tendency to rise. Creatine kinase in the tissues reveals several activity peaks. In plasma the activity course is U-shaped. Adenylate kinase in spleen and lung rises, whereas in muscle the activity varies considerably. The nearly identical decrease of alkaline phosphatase activity in the tissues during ageing is also reflected by a concomitant behaviour in plasma. Leucine arylamidase in lung and muscle both have a U-shaped characteristic, whereas in spleen the activity changes in a shorter period. In plasma, a rhythmical behaviour is apparent. Aldolase in plasma tripled during the observation period. Except for lactate dehydrogenase and aldolase, distinct sex-differences are observed in plasma. With progressive age the animals suffer increasingly from characteristic diseases, which beside experimental components have influenced the enzyme pattern. Enzyme activities in plasma and tissues show a complex pattern and are only of limited importance in understanding the ageing process.

Myocardial cell damage and breakdown of cation homeostasis during conditions of ischaemia and reperfusion, the oxygen paradox, and reduced extracellular calcium.

Enzyme release from perfused rat heart was determined under various conditions of injury. In analogous experiments, intracellular cation concentrations were measured using ion-selective microelectrodes. Under appropriate conditions, the inhibition of mitochondrial and/or glycolytic ATP production led to a decrease in the release of enzymes. During ischaemia or the oxygen paradox, the sarcosolic Ca2+ concentration was highly elevated; reperfusion or reoxygenation was followed by a drastic enzyme release. This was also found to be true under the conditions of an increased permeability brought about by a reduced extracellular Ca2+ concentration of 0.1 mmol/l. The intracellular pH under all conditions of injury was only moderately decreased. The sarcosolic Na+ concentration was markedly increased whereas the K+ concentration was decreased. The critical Ca2+ concentration of the sarcosol beyond which cell damage and enzyme release are inducible was assumed to be in the range between 10 and 32 mumol/l. The driving force of the Na+/Ca2+ exchange reaction of the sarcolemma is discussed in relation to recovery from hypoxic injury and the potential for avoiding cell damage.

Catalytic enzyme activity concentration in tissues of man, dog, rabbit, guinea pig, rat and mouse. Approach to a quantitative diagnostic enzymology, III. Communication.

The catalytic activity of up to fifteen enzymes was investigated in the liver, heart, skeletal muscle, kidney (medulla, cortex), brain, lung, duodenum, spleen and pancreas from man and animals. Human specimens were obtained from autopsies and immediately post-mortem from dogs, rabbits, guinea pigs, rats and mice. The differences between our results and previous reports of considerably lower activities for structural enzymes (e.g. creatine kinase) and for enzymes partly of mitochondrial origin (e.g. glutamate dehydrogenase, aspartate aminotransferase, malate dehydrogenase), is attributed to our use of a detergent extraction technique. The superiority of the detergent technique with regard to enzyme yield is exemplified by a comparison of various methods of extraction in rat liver, heart and skeletal muscle. Use of standardized assays allows a qualitative inter-species comparison of results. The influence of autolysis on catalytic activity of human autopsies is considered of minor importance.

[Quality management in the heart catheterization laboratory]. / Qualitätsmanagement im Herzkatheterlabor.

Quality management within the catheterization laboratory includes the quality control, the heart catheterization technique and the policy. Quality management is critical in the heart catheterization laboratory. Dedication of all members of the lab and computer personnel ensures high patient satisfaction. A continued quality improvement program is patient-orientated and requires good planning. One of the main emphasis in the catheterization lab is standardization which includes the patient preparation, the procedure itself, and the management. It is supported by teamwork including the economic aspect of prompt delivery of material and avoidance of complications. A continuous circle of treatment planes, performance, and check is regarded as the Deming cycle and leads to continuous improvement of quality. Important are both the avoidance and detection of complications. The reasons for any such have to be evaluated. It is recommended to follow the zero mistake hypothesis of Crosby, which means quality control by the lab supervisor, a quality consciousness, a quality measurement and quality improvement, as well as using a day to day quality improvement and to teach quality control. In Germany a quality control questionnaire was administered in an analysis of the current structure, function, and results of catheterization labs. Most important was the analysis of complications. The data were based on diagnostic catheterization in 1992, which included 140668 catheterizations in 83 laboratories. Thus, a mean of 1030 heart catheterizations was performed in each lab. In the mean, 200 catheterizations were performed by each doctor. In 19% of the labs digital imaging was exclusively performed. Major complications occurred with ventricular fibrillation in 0.36% (range 0.75%), resuscitation 0.18% (0.43%), persistent cerebrovascular accident 0.08% (0.24%), myocardial infarcts 0.19% (0.59%), aortic dissection 0.05% (0.22%). Mortality was 0.03% (0.08%). In heart catheterization laboratories quality management is one of the major goals for the future work. Only the continued improvement of quality and very good quality management ensure patient safety. Quality is the sum of technique and consciousness.

Evaluation of granulocyte-releasing products and chemiluminescence during cytapheresis.

During the procedures of centrifugation leukapheresis and plateletpheresis, donors occasionally experience adverse clinical reactions. The possibility of whether the activation of granulocytes and the subsequent release reactions, which may have been triggered by this extracorporeal circuit, were responsible for these adverse effects was evaluated. Six blood samples were obtained at set intervals during cytapheresis. Of these samples, four were taken directly from the donor. The remaining two were drawn from the efferent lines, i.e., those which return blood from the cytapheresis machine to the donor. Reactive oxygen species produced by granulocytes were measured by chemiluminescence (CL) using microamounts of whole blood or isolated granulocytes. Furthermore, secreted granulocyte products such as neutral proteinase elastase, which is present in plasma in a complex with alpha-1-proteinase inhibitor (E-alpha-1-PI), and lysosomal beta-glucuronidase were examined. A complete blood cell count and the values of hemoglobin, hematocrit, lactate dehydrogenase, protein, albumin, and proteinase inhibitors such as alpha-2-macroglobulin and alpha-1-proteinase inhibitor were also determined. Clinical chemical and cytologic values, with the exception of those for E-alpha-1-PI, were 10 to 17 percent lower than values before apheresis. These results can be attributed to inherent plasma volume expansion. Reduced CL was observed on the stimulation of phagocytes in the whole blood assay, as well as with stimulated granulocytes. Unstimulated granulocytes, on the other hand, showed an increased native CL. These data do not indicate a cytapheresis-mediated activation of the oxidative metabolism of granulocytes, and the concomitant discharge of proteolytic enzymes remains, therefore, of no clinical importance.

Enzyme activities in blood cells of man and dogs after separation on a discontinuous Percoll gradient.

From human and dog peripheral blood, thrombocytes (TRC), mononuclear cells (MNC), polymorphonuclear cells (PMNC), and red blood cells (RBC), were harvested after separation on a discontinuous Percoll gradient in a two-step centrifugation procedure, and 12 enzyme activities were determined in these highly purified cells. The enzyme activities measured are generally severalfold higher than previously reported, a fact which is attributed to the gentle and time-shortening isolation in Percoll, the cell disruption technique using detergent and the enzyme test conditions.

Enzyme activities in rabbit, guinea pig, rat and mouse blood cells after separation on a discontinuous Percoll gradient.

A technique is described which allows a fractionation of rabbit, guinea pig, rat and mouse blood cells using a discontinuous Percoll gradient. With only two steps of centrifugation a simultaneous isolation of thrombocytes, mononuclear cells, polymorphonuclear cells, and erythrocytes in nearly pure form is performed in a very short time, starting from only 1 ml of blood. In these morphologically almost homogeneous cell fractions the activities of 12 enzymes were determined. The enzyme pattern, which with respect to the number of enzymes, specific cell populations and species was not yet investigated to such an extent, revealed a manifold higher enzyme content in the few cases, in which comparative studies were made. This can be attributed mainly to the completeness of the cell disruption technique using a detergent and certainly also to the use of Percoll as a preferable gradient material.