Animal model of experimentally induced ectopic enchondral ossification in nude mice with xenograft of porcine epiphyseal cartilage.

In a specific animal model the induction of heterologous enchondral ossification was examined in long term follow-up. Physiologically normal and devitalized frozen porcine epiphyseal cartilage as a control group were transplanted as s.c. xenograft into athymic nude mice (nu/nu). 15 weeks after transplantation the porcine growth plate cartilage showed the development of viable cartilage tissue in the recipients, further differentiating to juvenile long bone with epiphysis, diaphysis and growth plate. Porcine growth plate cartilage increased s.c. in the recipient nude mice, showing enchondral ossification without any physiological load, and finally an unexpected development towards complete juvenile long bone.

Dose-dependent induction of transforming growth factor beta (TGF-beta) in the lung tissue of fibrosis-prone mice after thoracic irradiation.

PURPOSE: The lung is the major dose-limiting organ for radiotherapy of cancer in the thoracic region. The pathogenesis of radiation-induced lung injury at the molecular level is still unclear. Immediate cellular damage after irradiation is supposed to result in cytokine-mediated multicellular interactions with induction and progression of fibrotic tissue reactions. The purpose of this investigation was to evaluate the acute and long-term effects of radiation on the gene expression of transforming growth factor beta (TGF-beta) in a model of lung injury using fibrosis-sensitive C57BL/6 mice. METHODS AND MATERIALS: The thoraces of C57BL/6 mice were irradiated with 6 and 12 Gy, respectively. Treated and sham-irradiated control mice were sacrificed at times corresponding to the latent period (1, 3, 6, 12, 24, 48, 72 hours and 1 week postirradiation), the pneumonic phase (2, 4, 8, and 16 weeks postirradiation), and the beginning of the fibrotic phase (24 weeks postirradiation). The lung tissue from three different mice per dosage and time point was analyzed by a combination of polymerase chain reaction (PCR), immunohistochemistry, and light microscopy. The mRNA expression of TGF-beta was quantified by competitive reverse transcriptase/polymerase chain reaction (RT-PCR); the cellular origin of the TGF-beta protein was identified by immunohistochemical staining (alkaline phosphatase-anti-alkaline phosphatase [APAAP]). The cytokine expression on mRNA and protein level was correlated with the histopathological alterations. RESULTS: Following thoracic irradiation with a single dose of 12 Gy, radiation-induced TGF-beta release in lung tissue was appreciable already within the first hours (1, 3, and 6 hours postirradiation) and reached a significant increase after 12 hours; subsequently (48 hours, 72 hours, and 1 week postirradiation) the TGF-beta expression declined to basal levels. At the beginning of the pneumonic phase, irradiation-mediated stimulation of TGF-beta release reached maximal values at 2 and 4 weeks. The elevated levels of TGF-beta mRNA during the latent phase have been found to correlate with immunohistochemical staining of alveolar macrophages. The most striking increase in TGF-beta immunoreactivity was seen during the acute phase of pneumonitis. Throughout this observation period, type II pneumocytes and fibroblasts (apart from inflammatory cells) served as important sources of TGF-beta expression. Increased TGF-beta expression was detected prominently in regions of histopathologic radiation injury. After exposure to a single radiation dose of 6 Gy, the lung tissue revealed only a minor radiation-mediated TGF-beta mRNA response. The modest upregulation ranged from 6 hours to 48 hours after irradiation. Corresponding to the only minor histopathologic changes after thoracic irradiation with 6 Gy, measurement of TGF-beta mRNA levels during the later time points revealed no significant alterations in comparison to untreated control mice. CONCLUSIONS: This study demonstrates an acute and long-lasting increase in the expression of TGF-beta in lung tissue following thoracic irradiation with 12 Gy. The predominant localization of TGF-beta in areas of inflammatory cell infiltrates and fibrosis suggests involvement of this cytokine in the pathogenesis of radiation-induced pulmonal fibrosis. Further studies should be performed to explore the role of other cytokines in the development of radiation injury. An improved understanding of the underlying mechanisms of pulmonary fibrosis may eventually lead to modulatory intervention at the molecular level to modify the fibrotic process.

Bactericidal effect of extracorporeal shock waves on Staphylococcus aureus.

Despite considerable knowledge about the effects of shock waves on eukaryotic soft tissues, no data are available concerning their effect on prokaryotic micro-organisms. In vitro studies on the bactericidal effect of extracorporeal shock waves on staphylococci were performed with energy levels that are standard for the disintegration of calculi. Suspensions containing 10(4)-10(5) cfu of Staphylococcus aureus/ml were sealed in plastic tubes and exposed to shock waves, resulting in a mean decrease of 3.1 log(10). Whereas impulse rates of > or =350 resulted in a decrease of cfu/ml equalling the detection limit, lower numbers of impulses did not result in an appreciable bactericidal effect. The bactericidal effect of extracorporeal shock waves might provide the basis for the development of novel therapeutic strategies for bacterial infections.

Estradiol and high-dose dihydrotestosterone treatment causes changes in cynomolgus monkey prostate volume and histology identical to those caused by testosterone alone.

To evaluate the effects of testosterone (T), dihydrotestosterone (DHT), and estradiol (E2) on the regulation of prostate growth and tissue composition, the following study was conducted in a nonhuman primate model. Fifteen adult, long-term castrated cynomolgus monkeys were randomly assigned to receive implants filled with T (0.19 +/- 0.01 g), DHT alone (0.21 +/- 0.01 g), or (99%) DHT + (1%) E2 (0.21 +/- 0.01 g). Prior to and at 4-week intervals during the treatment phase of 252 days, prostate volumes (PV), body weight, ejaculate weight, hormone levels (of T, DHT, and E2), and red blood cell count were measured. Five adult, intact monkeys served as controls for prostate volume and histology. At the end of the study, histological analysis of an ultrasound-guided prostate biopsy was performed. T levels increased significantly in the T group compared with baseline (P < 0.01) and with the DHT and DHT + E2 groups (P < 0.05). Both groups receiving DHT showed higher DHT levels than did animals in the T group (P < 0.001). E2 levels in all groups increased over time (P < 0.05), although significant differences (P < 0.01) could only be detected between the DHT + E2 and the DHT group. Prostate volume in all groups increased (at baseline: T = 1.03 +/- 0.12 ml, DHT = 1.08 +/- 0.15 ml, DHT + E2 = 1.13 +/- 0.09; at day 252: T = 5.83 +/- 1.00, DHT = 4.72 +/- 0.9, DHT + E2 = 5.05 +/- 0.62) over time (P < 0.001), whereas no differences could be detected between the groups. Prostate biopsy could be performed successfully in 15 out of 20 monkeys. Prostate tissue evaluation between the treatment groups and the evaluated intact monkeys revealed no differences in the status of secretory epithelia, nuclear chromatin, excretory vacuoles, interstitial stroma, smooth muscles, and total functional status, whereas the prostate of a long-term castrated monkey showed severe atrophy. Thus, both androgens fully restored prostate volume and ejaculatory function. Highly supraphysiological DHT serum levels are not associated with abnormal volumetric or histological changes of the prostate. Comparing the DHT group with the DHT + E2 group, an additional stimulatory effect of normal or slightly elevated estrogens on the prostate cannot be found in the presence of highly supraphysiological DHT levels.

3 beta-Chlorosteroids: artefacts and possible contaminants in food and feed--toxicological effects in mice.

3 beta-Chlorosteroids, such as cholesteryl beta-chloride and sitosteryl beta-chloride, are formed during the production of protein hydrolysates, which are useful flavour enhancers. These chlorinated steroids may also attract attention as environmental contaminants if they are released from liquid crystal display devices. The effects of orally administered 3 beta-chlorosteroids were tested in female NMRI mice. The animals were fed cholesteryl beta-chloride or sitosteryl beta-chloride at doses of 1 mg and 10 mg/animal/day, that is, 33 mg and 330 mg/body weight/day, over a period of 3 months. Feed intake, body weight and organ weights of the animals, as well as concentration of 3 beta-chlorosteroids in faeces and various organs and tissues showed that cholesteryl beta-chloride and sitosteryl beta-chloride are not acutely toxic compounds. However, chronic toxicity cannot be excluded because small amounts of 3 beta-chlorosteroids, in particular cholesteryl beta-chloride, were absorbed by the intestinal tract and accumulated in adipose tissue. Histopathological examination of sections of organs and tissues showed no indication of irreversible cell damage in the stomach, duodenum, liver, kidneys and spleen caused by the chlorinated steroids.

[Uncemented total prosthetics as exemplified by a carbon-fiber prosthesis of the hip joint]. / Zur Problematik der zementfreien Totalprothetik am Beispiel einer Kohlefaserprothese des Hüftgelenkes.

A new artificial total hip joint was designed and tested in animals and human patients up to five years. For implant combined carbon material (Endocarbone) was used. The primary anchorage was achieved by a hydraulic pin system without bone cement. The follow-up of bone growth at the implant bone border line was proved by X-ray, computer-tomography, arthrography and histo-morphological tissues. The Endocarbone hip prosthesis has been proved as a biological implant. The good bonely incorporation justifies a more intensive clinical testing.

The heart muscle's putative "secondary structure'. Functional implications of a band-like anisotropy.

Opinions are divided as to whether the rope-like secondary structure, which Torrent-Guasp dissected out of the myocardial body by the blunt unwinding technique (BUT) reveals some kind of functional compartmentation of the heart muscle. The myocardial fibres are aligned parallel to the fibre disruption (cleavage) plane, along which the band has been prepared but they are not necessarily aligned parallel to the long axis of the band. Inconsistencies in the myocardial rope model arise from the obligatory zones of transmural inflection, which are obvious in the base and the apex of both ventricles. They are, however, merely discernible in the midzone of the left ventricular cone. The investigator experienced in BUT knows that the cleavage plane is not unique. We doubt the assumption that the rope structure is the predominant stress transmission pathway, because the fibre strand peel-off technique (SPOT) delivers irregular fibre disruption planes which are definitely different from those which Torrent-Guasp prepares. The rope-like fibre arrangement could be just a redundant structure, a remnant of past developmental steps without, however, any functional implication to the human heart. On the other hand, peeling-off fibre strands from the ventricular wall produces deeply perforating, i.e., oblique transmurally grooved surfaces. Putative functions of force transmission in an oblique transmural direction are (1) ventricular dilation as a function of the variable inclination angle with respect to the epicardial surface, (2) monitoring of ventricular wall stress and ventricular size and (3) segmental stiffening which could serve other dependent segments as a punctum fixum.

The heart's fibre alignment assessed by comparing two digitizing systems. Methodological investigation into the inclination angle towards wall thickness.

Myocardial contractile pathways which are not aligned strictly parallel to the heart's epicardial surface, give rise to forces which also act in the ventricular dilating direction. We developed a method which allows us to assess any fibre orientation in the three-dimensional myocardial weave. Decollagenized hearts were prepared by peeling-off fibre strands, following their main fibre orientation down to near the endocardium. In the subepicardium the strands followed a course more or less parallel to the epicardium, whereas from the mid-wall on they tended to dive progressively deeper into the wall. The preparation displays more or less rugged surfaces rather than smooth layers. The grooves and crests on the exposed surfaces were sequentially digitized by two methods: (1) Using a magnet tablet (3 Draw Digitizer System, Polhemus, Cochester VTO 5446, USA) on a dilated pig heart we manually followed the crests using a stylus, handling each groove and crest as an individual contractile pathway. (2) A constricted cow heart was digitized using a contact-free optical system (opto TOP, Dr. Breuckmann, Meersburg, Germany), which is based on the principle of imaging triangulation. Using specially developed software the inclination angles of selected crests and grooves with respect to the epicardial surface were calculated. The two digitizing methods yield comparable results. We found a depth- and side-specific weave component inclined to the epi-endocardial direction. This oblique netting component was more pronounced in the inner 1/3 of the wall than in the subepicardium. The inclination angle probably increases with increasing wall thickness during the ejection period. Manual digitizing is an easy and fast method which delivers consistent results comparable with those obtained by the cumbersome high resolution optical method. The rationales for the assessment of transmural fibre inclination are (1) the putative existence of dilating forces inherent in the myocardial weave and (2) the possible overproportional increase in the oblique transmural weave component during myocardial hypertrophy, which would entail a reduction in efficiency of ventricular performance in terms of haemodynamic work.

The assessment of intramural stress alignment on the beating heart in situ using micro-ergometry: functional implications.

The main local stress transmission pathways in the left ventricular base, midportion and apex in up to seven layers have been assessed in normal dog and porcine hearts, in hypertrophied dog hearts, and in three pig hearts having undergone a temporary left ventricular outflow stricture. The rotational sensitivity of needle force probes was used to determine the focal surface-parallel direction of the myocardial tension vector. In all places investigated the orientation of the force transmission pathways differs slightly from the morphologically determined fibre alignment. Vector rotation upon an axis normal to the epicardial surface is definitely tempered as compared to fibre rotation. Alterations in the force transmission pathways assessed in hypertrophied dog hearts by micro-ergometry qualitatively confirm structural remodelling in so far as an irregularity in the transmural rotation of the main stress vector was found. The measured disparities between the alignment of the myocardial fibre weave and the direction of stress transmission both in the normal and the diseased heart is widely individual, and hence, scattering of the data is marked. However, it must also be called into consideration that the measured orientation of force vectors is that at the moment of highest developed force, only. Further investigations will elucidate if discrepancies between that force vector and morphology are less pronounced when the vector is averaged over the entire heart cycle.

The difficult access to morphology of the heart: clinical implications.

The literature on the morphology of the heart is reviewed within the context of recent histological findings. There is strong evidence for a dualistic myocardial function, whereby both ventricular constricting and expanding forces are supposed to act synchronously although with variable effect over the heart cycle.The morphological basis of this dualistic myocardial function is the contorted rope-like structure worn into the bulk of the heart muscle. Opinions are divided about the invasiveness of blunt preparation on the heat denatured heart by which the fascicular architecture is carved out of the muscle. Histology confirms the existence of a fascicular substructure. It results from an inhomogeneous repartition of myocardial fibre branchings and the arrangement of the delicate connective tissue netting by which myocardial strands are bundled and wrapped. One important feature of the fascicular structure of the heart muscle is an oblique transmurally arranged element which yields a force vector opposing systolic wall thickening. This structural element which acts in the direction of ventricular dilation probably gains pathological import in some cases of architectural remodelling, namely in myocardial fibrosis and hypertrophy.

[Direct radiographic magnification in bone infections]. / Direktradiographische Vergrösserung bei Knocheninfektionen.

An experimental model of bacterial osteomyelitis was used to investigate the correlation of direct radiographic magnification and histopathologic changes in the femora of rabbits. The histopathologic changes demonstrated a severe chronic state of osteomyelitis. The extent of radiographic changes was more clearly recognizable with direct radiological enlargement (up to 15-fold) than on normal radiographs: this applied to destruction of marrow and cortical bone, formation of sequestra, and (frequently) extraosseous extension. Histopathologic findings were in agreement with the changes revealed by direct radiographic enlargement.

The protective effect of Allium sativum and crataegus on isoprenaline-induced tissue necroses in rats.

Possible protective effects of Allium sativum and Crataegus--alone and in combination--on isoprenaline (isoproterenol)-induced heart, liver and pancreas damage were studied using rats as test animals. Pretreatment with Allium sativum alone, or in combination with Crataegus, resulted in protective effects on isoprenaline-induced damage of heart, liver, and pancreas. These effects proved to be dose-dependent. The following parameters were used to evaluate the protective effect: Clinical signs, qualitative histological and histoenzymatical findings, as well as quantitative microphotometric determination of enzymatic activities of succinate dehydrogenase, NADH-NBT reductase, acid phosphatase and glucose-6-phosphate dehydrogenase in cardiac, hepatic and pancreatic tissues. The underlying mechanisms are discussed. The results suggest that Allium sativum, resp. Allium sativum plus Crataegus exert a pronounced protective effect.

[Cement-free fixation of a stemless total knee endoprosthesis in animal experiments (author's transl)]. / Zur Problematik von schaft- und zementfreien Totalprothesen des Kniegelenkes--eine tierexperimentelle Studie.

A stemless total knee endoprosthesis with physiological kinematics (slip and rolling motion, tibial rotation) and cement-free primary fixation (straddling dowel and thread system) was tested in animal experiments. The fixation allows total functioning of the operated extremity from the first day after operation. Primary results and follow-up for 40 weeks after operation reveal satisfactory results, especially with regard to durable bony incorporation of the prosthesis as shown by histologic and microangiographic studies.

Fat infiltration in liver of rats induced by different dietary plant oils: high oleic-, medium oleic- and high petroselinic acid-oils.

Beneficial effects on health parameters by ingestion of plant oils having high levels of monounsaturated fatty acids such as oleic (cis-9-octadecenoic) acid have been described in several studies. Recently, we have shown that feeding of coriander oil containing high proportions of a positional isomer of oleic acid, i.e. petroselinic (cis-6-octadecenoic) acid, led to significantly decreasing proportions of arachidonic (all cis-5,8,11,14-eicosatetraenoic) acid in the cellular lipids. Here, we report histopathological findings in male Wistar rats fed isocaloric diets containing 2% corn oil and 12% (w/w), each, of different plant oils (high-oleic and conventional sunflower oil, olive oil, rapeseed oil as well as coriander oil) containing varying levels of monounsaturated fatty acids, i.e. oleic or petroselinic acids. One group of animals was fed a standard diet containing 4% (w/w) of fat as control. Liver, heart, aorta, stomach, and spleen were anatomically and histologically examined. Livers of animals from all the groups that had received high-fat diets-except the coriander oil group-showed moderate non-degenerative fat infiltrations of the hepatocytes beginning in the periportal fields. Marked to severe fat infiltration was observed in hepatocytes of animals fed coriander oil. In this experimental group the livers were found to contain fatty cysts in addition to hepatocytes with mixed-size lipid vesicles. Moreover, enlarged nuclei were observed in numerous hepatocytes without fat infiltration. Fat infiltration was not observed in livers of animals fed a rat standard diet. In the cells of all other organs and tissues studied, in particular heart and aorta, neither non-degenerative fat infiltrations nor other degenerative changes were observed for any group. Our results show that feeding of fat-rich diets containing varying proportions of C18 cis-monoenoic fatty acids induced different histopathological alterations in the livers of rats, depending on position of the double bond. Plant oils containing varying proportions of oleic acid lead exclusively to moderate fat infiltration in the hepatocytes, whereas in the animals fed high-petroselinic coriander oil degenerative alterations (fatty cysts) as well as -in several cases-enlarged nuclei were found that are likely due to a non-specific stimulation of the hepatocytes resulting from the cell degeneration observed.

[Comparative study of the detection of E. coli after time in seawater with partially modified standard media]. / Vergleichende Untersuchung zum Nachweis von E. coli nach Aufenthalt in Meerwasser mit partiell modifizierten Standardmedien.

Fecal coliform bacteria (E. coli) are the most important parameter when monitoring the hygienic quality of bathing water according to EC guidelines. Among other things the number of E. coli is clearly dependent on the length of exposure of the bacteria to marine water as well as the media used to establish the number. In a seven-day salt water stress experiment the best rate of recultivation was achieved on average with DEV lactose peptone broth. The Brila-MUG medium produced a slightly lower rate of recultivation and the fluorocult-lauryl-sulfate-broth the lowest rate of recultivation. After a shorter exposure of bacteria to marine water conditions (up to 24 h) the DEV-lactose-broth produced slightly lower rates of recultivation. An alteration of the standard components of the media did not lead to an improvement in the rate of recultivation.

[Thorotrastosis--a rare disease picture in internal medicine]. / Die Thorotrastose--ein seltenes Krankheitsbild in der Inneren Medizin.

The report deals with the late effects following administration of thorotrast in a 63-year-old patient: liver cirrhosis developed after application of thorotrast 25 years ago. The clinical findings are described and attention is drawn to the relevance of this disease at present time. The authors refer to the significance of the routine X-ray diagnosis and of the CT for finding a diagnosis with thorotrastosis.

Effect of extracorporeal blood irradiation on lymphocytes and the lymphatic tissue in a goat.

In a long-term study discontinuous extracorporeal blood irradiation (ECIB) was applied to a goat using a 500 Ci-137 Caesium source. Lymphocytes of the peripheral blood were examined by light and electron microscopy. After application of a transit dose of 466,500 rad the lymphocytes in the peripheral blood were found to be decreased from 6,900/microliter to 500/microliter, revealing a complete dissolution of the nuclei in electron microscopic preparations. Histological examinations showed a severe atrophy of the whole lymphatic tissue.

[Experimentally induced pancreatitis (author's transl)]. / Experimentelle Untersuchungen zur Pankreatitisentstehung.

Pancreatitis was produced in 29 mini-pigs by partial occlusion of the pancreatic duct; occlusion was performed by implanting a T-drain into the duct. Inflammatory changes were induced additionally by applying radiopaque material in excess. Acute and chronic pancreatitic lesions could be diagnosed by X-ray examination, evaluation of blood chemistry and by histology.

[Acute pancreatitis in animal experiments - a new method]. / Die akute Pankreatitis im Tierexperiment - eine neue Methodik.

Acute pancreatitis was induced in 30 minipigs (Göttingen) by implantation of a T-drain into the Ductus Wirsungianus. The distal part of the T-drain, which led into the duodenum was stenosed to 1/3 of the original diameter. The inflammation was caused by stenosis of the Ductus Wirsungianus additionally by application of contrast medium. The course of the inflammation was demonstrated by biochemical, roentgenological and histological examinations.