Size dependent enzyme activities of multinucleated (osteoclastic) giant cells in bone tumors.

In osteoclastic giant cells of six different tumors of bones and joints (fibrous dysplasia, proliferating giant cell tumor, malignant giant cell tumor, osteosarcoma after chemotherapy, malignant synovioma and Ewing's sarcoma) activities of tartrate-resistant acid phosphatase, NADH-tetrazolium-oxidoreductase and, in three of them, of non-specific esterase are determined by enzyme histochemical methods. Quantitative microphotometry makes it possible to determine relative enzyme activities in the cut sections of giant cells of different sizes. Giant cells of the various tumors reveal similar trends: With an increase in cell size, mean extinctions of NADH-tetrazolium-oxidoreductase and non-specific esterase decrease. Mean extinctions of tartrate-resistant acid phosphatase increase in cells of medium size, whereas the large cells reveal in part low activities. An additional ultrastructural examination of the giant cells in the proliferating giant cell tumor as well as in the osteosarcoma shows morphological signs of degeneration in the large cells. Electron probe microanalysis of the proliferating giant cell tumor exhibits evidence of phagocytosis of Ca and/or Fe containing particles. The similar size dependent reaction pattern of enzymes in osteoclastic giant cells of different tumors favors the concept of a common histogenesis, i.e. a host reaction.

Microphotometric quantitation of enzyme activities in giant cell tumor of bone. A case study.

In a proliferating giant cell tumor of bone the activities of tartrate-resistant acid phosphatase (acPase) and of NADH-tetrazolium reductase were demonstrated by enzyme histochemical methods. Quantitative microphotometry made it possible to determine the relative enzyme activities per given volume unit in the cytoplasm of giant cells of several sizes. The activity of tartrate-resistant acid phosphatase increases with increasing cell size, whereas the activity of tetrazolium reductase will decrease in proportion. This coincidence of high acPase activity and low tetrazolium reductase activity in larger giant cells is interpreted as an expression of degenerative change.

[Results of "cross-over implantation of Palmaz-stents" with cross-over sheaths]. / Ergebnisse der "Cross-over Implantation von Palmaz-Stents" mit Cross-Over Schleuse.

PURPOSE: Presentation of technique of cross-over stent implantation of Palmaz stents using a cross-over sheath. MATERIAL AND METHODS: Between 1996 and March 1997 72 patients presenting symptoms of peripheral artery disease received 136 Palmaz stents, which were implanted in a cross-over technique with the aid of a cross-over sheath. RESULTS: As indications for stent implantation occurred: occlusion (n = 14), recoil (n = 11), distinctive calcification (n = 32), recurring stenosis/occlusion (n = 32), perforation (n = 3), dissection (n = 31), and aneurysms (n = 2). The cross-over technique with the aid of a cross-over sheath was successful in all 136 cases. Within the first 24 hours 3 acute reocclusions occurred (all femoral). For stents placed in the iliac artery the primary patency rate was 93.3% at 12 months and for stents placed in the femoral artery 72.0%. Furthermore, this techniques allows for simultaneous stenting of pelvic, femoral or lower leg arteries (n = 17). By using this technique, stenting of the distal external iliacal artery, common femoral artery (CFA) and proximal superficial femoral artery becomes either possible for the first time or easier. CONCLUSION: If Palmaz stents with a length of 2-7.8 cm should be implanted in the cross-over technique, the use of a cross-over sheath is recommended.

[PTLA (percutaneous transluminal laser angioplasty) recanalization of femoral artery stenoses/occlusions after Angio-Seal administration]. / PTLA-Rekanalisation von Femoralarterienstenosen/-okklusionen nach Angio-Seal Applikation.

PURPOSE: To demonstrate desobliration of Angio-Seal-induced femoral artery stenosis and occlusion by Excimer-laser assisted angioplasty and PTA. PATIENTS AND METHODS: The Angio-Seal hemostatic closure device was applied to the puncture site in 1500 patients after diagnostic or therapeutic coronary artery catheterisation. In 5 of 1500 cases symptoms of claudication occurred. Following the determination of the walking distance and ankle-brachial systolic pressure index (ABI) and diagnostic angiography, therapeutic percutaneous transluminal laser angioplasty (PTLA) and PTA was performed. RESULTS: 5 patients with acute symptoms of peripheral artery disease presented with superficial femoral artery occlusions (three cases) and high grade stenoses of the common femoral arteries (two cases). Angiographic and clinical improvement was achieved after PTLA/PTA in all five patients. The mean walking distance increased from 61 meters to 600 meters. The average ankle-brachial systolic pressure index increased from 0.40 to 0.82. CONCLUSIONS: PTLA/PTA is a satisfactory therapeutic method for femoral artery occlusion or high-grade stenosis following Angio-Seal application.

[Changes in the ultrastruct of the rat kidney after intoxication with organophosphates (author's transl)]. / Uber Veränderungen an der Ultrastruktur der Rattenniere nach Phosphorsäureestevergiftung

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.

[Histochemical investigations on liver and kidney of the rat after intoxication with organophosphates (author's transl)]. / Histochemische Untersuchungen an Leber und Niere der Ratte nach Phosphorsäureestervergiftung

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 flow cytometry for diagnosis of male fertility.

The patient came with a severe testicular disturbance (oligoasthenoteratozoospermia) into the department of andrology. After antiphlogistic- and antiestrogen therapy the clinical spermatological findings (spermiograms) were compared with the corresponding histograms of the flow cytometry analysis and interpreted as according with them.

[The reactive behaviour of the cell nucleus after a pretreatment with a bromide solution and the application of the calcium-cobalt-method for the ATPase evidence (author's transl)]. / Das färberische Verhalten der Zellkerne nach Bromwasservorbehandlung und Anwendung der Calcium-Cobalt-Methode zum ATPaseNachweis.

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.

[Viable cells from human renal cell carcinoma: isolation procedure and analysis of hormone sensitivity (author's transl)]. / Vitale Nierenkarzinomzellen: Isolierungsmethode und Untersuchung der Hormonempfindlichkeit.

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.

[Differentiation of human sperm cells using flow cytometry]. / Differenzierung menschlicher Spermazellen mit Hilfe der Durchflusszytometrie.

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 the preparation of histological sections for electron microscopy]. / Eine verbesserte Methode zur Herstellung histologischer Schnitte für die Elektronenmikroskopie

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.

[Phosphoric acid ester poisoned rats after antidote therapy. 2. Determination of serum enzyme activity]. / Befunde an Phosphorsäureester-vergifteten Ratten nach Antidottherapie. II. Aktivitätsbestimmungen einiger Serumenzyme.

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.

[Lipids of plasmamembranes and of mitochondria in rat liver after intoxication with paraoxon (author's transl)]. / Lipide der Plasmamembranen und der Mitochondrien aus Rattenleber nach Paraoxon-Intoxikation.

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.

Flow cytometric discrimination of human semen cells.

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.

[Findings following antidote therapy in rats intoxicated with phosphoracetic acid. / Histochemical and biochemical lipid demonstration in liver and kidneys (author's transl)]. / Befunde an phosphorsäureester-vergifteten Ratten nach Antidottherapie. I. Histo- und biochemische Fettdarstellung in Leber und Niere.

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.

Mg2+-ATPase activity in the rat testis and its correlation with the stages of spermatogenesis: a histochemical study.

The activity of Mg2+-activated ATPase was studied in the rat testis employing histochemical and microphotometrical methods. The enzymatic activity could be correlated with the stages of spermatogenesis. From stage XII onwards, the elongated spermatids showed a weak to moderate activity, which increased from the acrosomal phase to the maturation phase. The reaction product was located in the mid-piece and tail piece of elongated spermatids with a maximum intensity at stage VIII. Morphometric analysis revealed six distinct patterns, which could be correlated with the stages of spermatogenesis.

[Analysis of spermatozoa with flow cytophotometric determination of DNA]. / Die Analyse von Spermaproben mit Hilfe impulscytophotometrischer DNS-bestimmungen.

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.

[The effect of organophosphates on heart ATPase in the rat]. / Wirkung von Organophosphaten auf Herz-ATPasen von Ratten.

The effect of organophosphate intoxication with diisopropylfluorophosphate (DFP), paraoxon and parathion on the activity of some ATPases of rat hearts was examined. Biochemical and histochemical methods were employed to determine the activities of the transport ATPase, of the myofibres and mitochondrial ATPases using different doses and antidote application. The organophosphates studied here had an inhibitory effect on the activities of enzymes. Biochemical measurements showed that the ATPase activities are dependent upon the duration of poisoning, the dose of poison and the applied organophosphates. DFP caused the highest restriction in activity of the examined ATPases. The Ca2+-stimulated ATPase was more sensitive than the Na+/K+-ATPase to the application of organophosphates. The antidote therapy determined the grade of influence on activity. Combined treatments of organophosphate poisoning with atropine and obidoxime chloride in the high doses of 7.5 mg/kg and 12.5 mg/kg body weight i.m. proved to be the most effective against the organophosphate intoxication. Antidote combination of small doses and also single application of atropine or obidoxime chloride might cause a stimulation of activity of the Na+/K+-ATPase and a significant decline of the Ca2+-ATPase.

Electron microscopic evidence for deep invaginations of the lamina propria towards the seminiferous tubule lumen in a patient with varicocele.

Ultrastructural studies on biopsy tissue from the right testis of a 39-year old patient with varicocele revealed 2.5-5 microns thick invaginations of the lamina propria towards the lumen of the seminiferous tubules. These invaginations were of various lengths. The presence of invaginations was confirmed through examination of serial semi-thin sections. In some seminiferous tubules two neighbouring deep invaginations were joined together thus completely encircling and thereby separating the basal compartment, and in some cases even the adluminal compartment, of the seminiferous tubule. The invaginations were surrounded continuously by the basement membrane and contained collagen fibres, cell processes of myoid cells and in some cases also their nuclei.