Biomedical subjects
Publications and source records attributed to H Hettwer.
An improved method for differential staining and high-resolution flow cytometric measurement of human sperm cells is presented. Involving a mild pretreatment with citric acid/detergent and staining with DAPI, the new procedure provides excellent preservation and good discrimination of all cells present in normal and pathologic semen samples.
An improved method for differential staining and high resolution flow cytometric measurement of human semen cells is presented. Using a mild pretreatment with citric acid/detergent and staining with DAPI, the new procedure provides excellent preservation and good discrimination of all cells which are present in normal and pathological semen samples.
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An enzymatic procedure for the isolation of metabolically active tumour cells from human renal cell carcinoma is described. The cells were suspended by a multistep incubation procedure of the tissue in the presence of collagenase (10 mg enzyme/g tumour wet weight) dissolved in a calcium-free buffer solution. About 90% of the isolated tumour cells were viable, as judged by routine trypan blue staining. Electron microscopic examination revealed tumour cells in various stages of dedifferentiation. The cells had retained their capability of protein synthesis. In short term experiments the effects of 17 beta-oestradiol and progesterone on the incorporation of [U - C] L-leucine into cellular proteins was studied; Progesterone was found to exhibit a slight tumour antianabolic or catabolic action. A 17 beta-oestradiol-dependent modulation of the rates of protein synthesis was not observed.
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Paraoxon in doses of one LD50 (0.426 mg/kg), eight times and eighty times LD 50 was applied s.c. to female Sprague-Dawley rats. After 3, 6, 10, 24 and 36--48 h the activities of enzymes GOT, GPT, GLDH, SDH, CPK and ChE were measured, once after i.m. antidote application of Toxogonin only, of Toxogonin + atropine and the next one after application of combination Toxogonin + atropine + Solcoseryl (low-molecular components of deproteinized blood from young calves. The values obtained showed that in spite of treatment with Toxogonin or Toxogonin + atropine the activities of the enzymes increased; this enhancement could be prevented by addition of Solcoseryl to Toxogonin + atropine. The ChE-activity after 36 h was equivalent to that of the control value. The effect of paraoxon in the initial phase of poisoning was discussed in connection with hypoxia and acidosis resulting from a respiratory insufficiency as well as the inhibition of ATPase-activity with restriction of the energy metabolism following: consequently the effect of Solcoseryl was interpreted as an activation of the disturbed energy metabolism.
0.426 mg/kg Paraoxon was applicated to female Sprague-Dawley rats. 4, 10 and 24 h after poisoning, cholesterol, glyceride and phospholipid content was measured in liver plasma membranes and mitochondria. After 4 h, cholesterol content of plasma membranes was significantly lower. Phospholipid content of plasma membranes was significantly higher than control values at all times of investigation, contents of neutral lipids and glycerides were also higher, but not significantly. In mitochondria, no significant difference of cholesterol content could be detected. Glyceride content was lower after 4 and 10 h, but phospholipid content higher at all experimental times. Differences found in cholesterol and phospholipid content of plasma membranes are discussed also in view of higher serum GOT, GPT and CPK activities known from other investigations. The differences in cholesterol and phospholipid content, their degree and their timing, which agrees with the timing of enzyme leakage, suggest that lipid composition of the plasma membrane may be one of the parameters which determine membrane permeability after organophosphate poisoning.
In order to improve or supplement the currently used therapy of poisoning with phosphoric acid esters, a protein free extract from calf blood containing low-molecular subtances (Actihaemyl, Solcseryl) has been tested in female Wistar rats. After poisoning with paraoxon, DFP and soman, a combination of Solcoseryl, Toxogonin and atropine sulfate prolonged the time until exitus, improved the overall rate of survival and considerably shortened the phase of regeneration of the experimental animals even when they had been intoxicated with high doses of poison.
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The fatty degeneration of the liver and kidney of Sprague-Dawley rats was observed by histochemical and biochemical means after intoxication with LD50 and 8 X LD50 O,O-diethyl-O-p-nitro-phenylphosphate (paraoxon). The degree of fatty degeneration in these organs was demonstrated alternately after antidote-treatment with bis(pyridinium-4-aldoxim-1-methyl) ether (Toxogonin) and after injection of a combination of Toxogonin, atropine and a protein-free extract from calf blood containing low-molecular substances (Actihaemyl, Solcoseryl). Under the influence of Actihaemyl the degree of fatty degeneration induced by the toxin was, in respect to both triglycerides and phospholipids, near control levels. The mode of action of Actihaemyl is discussed with regard to the activation of fatty acids.
A selective staining of the nucleus of both the cells in an islet (of Langerhans) and the acinar cells could be observed after treatment with bromide-water in an ATPase-medium (calcium-cobalt-method) on a cryostatsection of rat pancreas. Furthermore the behaviour of the other metal ion forming a complex has been tested in place of cobalt salt. As conditioned for the staining reaction the hydrolysis of nucleic acid with oxidation of its base by bromide-water and also the complex formation of Co++ with compounds similar to alloxan, resulted from oxidation were discussed.
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Unsatisfying cut and electronmicroscopical figures of liver- and kidney preparations after organophosphate poisoning with Vestopal W, could be removed by experiments with new mixtures consisting of Vestopal W and Mikropal, obtaining reproductable cut-thickness and contrasty pictures for electronmicroscopical exploration.
Comparative histochemical investigations of the livers and kidneys of female Sprague-Dawley rats were made: 1. after intoxication with O-pinacolyl-methyl-phosphonylfluoride (Soman), O,O-diethyl-O-p-nitrophenylphosphate (Paraoxon, E 600); 2. after starvation of 24 to 36 h; 3. after hypoxia for 24 h. From the results it is concluded that the fatty degeneration of liver and kidney after PE.-intoxication is caused by intracellular hypoxia. No fatty degeneration of the organs was observed following deprivation of food for 36 h. A decreased level of cholin which might be caused by the intoxication with PE., and which could be the reason for the fatty degeneration was not found. This was indicated by the unaltered evidence of phosphatids in the intoxicated animals.
The kidneys of female Sprague Dawley rats were examined by electronmicroscopical and histochemical methods after intoxication with O-pinacolyl-methyl-phosphonylfluoride. The same explorations were made with control animals, with animals after hypoxia for 24 h, and starving for 36 h. The epithelial cells of the proximal and distal convoluted tubule cells showed the following changes: In the epithelial cells of the basal labyrinth there was a striking extention. The swelling of the intercompartmental space were discussed in connection with a restricted return of sodium-resorption. Further changes in the epithelial cells of the two nephron-segments after organophosphat-intoxication are the swelling of the mitochondria, the reduction of the cristae up to the total destruction of these mitochondria, the formation of myelin-figures, the swelling of the ER and the GOLGI complex.
Semen samples of fertility patients and fertile men were analysed by an improved technique of flow cytophotometry. The DNA-contents of mature spermatozoa were measured with a high level of accuracy. Besides spermatozoa, peaks of spermiogenetic cells and other seminal cells could be demonstrated. Complementary to the results of semen analyses the complex histograms of pulse cytophotometry investigations may give further informations on the nature of the underlying disturbances of spermatogenesis.