E. Somogyi

The ultrastructural demonstration of mitochondrial ATPase activity by the ATP-dependent accumulation of Ca++.

SummaryA new method was developed for the ultrastructural demonstration of mitochondrial ATPase activity based on the ATP dependent mitochondrial accumulation of Ca++. Reaction product labels the inner mitochondrial compartment in the form of electron dense precipitate granules. Reaction could be blocked with 10 mM oligomycin.

On the possibilities of application of scanning electron microscopy in the forensic medicine

SummaryThe aim of the paper is to provide pertinent, information about scanning electron microscopy (SEM), with a detailed survey on the great possibilities of SEM which is becoming increasingly important in research and practical work of forensic medicine. The electronoptical characteristics of the method are discussed and the basic preparation methods to be selected are described. The areas of forensic medicine where these mothods have already been used, as well as the results obtained, are briefly surveyed. The present state of affairs as well as personal experiences with hairs, bones, muscle and skin are described in detail. The experience with the critical point drying method is described. This method according to the reviewers, is very useful for work with hairs, bones, nails and sometimes with skin although the preparation may result in secondary destructions of the tissues of high water content to such an extent that the evaluation will be interfered with or becomes impossible. Further possibilities of these perspectivic methods are under research. The discussion of the physiological data is preceded by a historical description of the SEM and TEM systems and the basic principles of its function, which should facilitate reading of the text.ZusammenfassungDie Autoren geben eine ausführliche Übersicht über die Möglichkeiten von Rasterelektronenmikroskopie (REM), die in der Forschung und praktischen Arbeit der gerichtlichen Medizin eine zunehmende Bedeutung und ein immer wachsendes Applikationsgebiet aufweist.Die Grundlagen der Elektronenoptik und die Präparationsmethoden, die nach den verschiedenen Typen des Untersuchungsgutes variieren, werden beschrieben.Eine Übersicht über den bisherigen Anwendungsbereich der Methode im Fach der gerichtlichen Medizin wird gegeben.Die eigenen Ergebnisse, hauptsächlich bei der Untersuchung von Haaren, Knochengewebe, Muskelgewebe und Haut, werden ausführlich dargestellt. Auch über die eigenen Erfahrungen mit der „critical point Drying“-Methode wird berichtet. Diese Methode bewährt sich nicht bei der Bearbeitung von Haaren, Knochen und Nägeln, fallweise auch mit Hautgewebe. Bei Geweben mit hohem Wassergehalt kann die Präparation sekundäre Destruktionen bedingen, die die Auswertung stören oder fallweise sogar verhindern können. Die weiteren Möglichkeiten dieser perspektivischen Methode werden erörtert.

Cytochemical demonstration of the molecular forms of cardiac glycosides in the hart muscle

Selective topo-optical staining of vicinal-OH groups with aldehyde-bisulphite-toluidine blue (ABT) has been used for studying the localization and molecular structural order of cardiac glycoside in heart muscle. The glycoside has strong metachromatic basophilia and negative birefringence. In addition to confirming the electron microscopic suitability of the ABT reaction has offered also ultrastructural evidence for the reliability of the topo-optical method. The localization appeared to be in extracellular connection with the outer side of sarcolemma membrane, internal surface of endothelial cell, and in the subsarcolemmal cysterne. The findings suggest that the topo-optical reaction is suitable for molecular analysis of cardiac glycoside.

Electron-microscopic histochemical study of myosin ATP-ase activity

SummaryThe sites of calcium-activated myosin ATP-ase reaction have been determined by a method modified from that of Padykula and Hermann. On coherent muscle preparations the enzyme was found to be mainly active in the segment A, although reaction was sometimes observed in the strip Z also. In several cases the precipitate appeared in a spherical shape in the segment A. Since no such phenomenon was observed in isolated bundles of myofibrils it is supposed to be an artefact. The reaction appeared in fibre bundles at 300 to 400 Å intervals, and a certain spatial periodicity was observed also on isolated thick filaments.

Application of a vascular graft material (Solcograft®-P) in experimental surgery

The implantation and post-implantation behaviour of a Solcograft-P vascular prosthesis in the aortic, aorto-iliac, carotid and vena caval positions in dogs was studied up to 100 d post-surgery in order to assess the suitability of this vascular material for use in man. Solcograft-P is prepared from the carotid arteries of calves by crosslinking the collagen stroma using adipyl dichloride. During the postoperative follow-up period of 3 month, 100% of the aortal grafts, 80% of the aorto-iliac bypasses, 60% of the vena caval grafts and 35% of the carotid implants remained patent. The biochemical properties of the Solcograft-P are better than those of Solcograft, its predecessor. The intimal lining was consistently smooth and homogeneous in grafts of biological origin, and no aneurysm was observed. Infection and early thrombosis occured no more frequently than with other grafts. The new Solcograft-P, crosslinked via ester and amide groups, seems to represent a real improvement over Solcograft. Our results suggest that Solcograft-P should prove valuable in various cases of reconstructive vascular surgery of the lower limb, especially when the autologous vena saphena magna is not available, and its mechanical properties may well prove suitable for both arterial and venous replacement.

Elektronenmikroskopische Untersuchung von Herzmuskelveränderungen, die unter dem Einfluß von Niederspannungs-Schrittmacher-Strom (Pacemaker) zustande gekommen sind

SummaryLow voltage electricity causes changes which may be divided into two groups:1. Actin and myosin damage in the primary and secondary fibers, predominantly in the A-segments.2. Changes due to hypercontractional relaxation of the fiber. These may include tearing of the fibers. In this group destruction of the mitochondria corresponds to morphologic changes in density. Electronmicroscopic histochemical methods enable characteristic localization of activity alterations.In the first group there are no changes visible by light microscopy but they are clearly evident in the second group.The changes suggest a relationship to the flow of current.ZusammenfassungDie Verfasser untersuchten unter dem Einfluß von elektrischen Niederspannungsstrom entstandene Muskelverä nderungen. Die Veränderungen unterteilten sie in 2 Gruppen:In der 1. Gruppe wurde eine Schädigung der Actin und Myosinkomponenten der primären und sekundären Filamente beobachtet, in erster Linie dem A-Segment entsprechend. Die 2. Gruppe der Veränderungen halten die Verfasser für eine Folge der Hyperkontraktion und Relaxation der Fasern, bei der auch das Zerreißen der Fasern eintritt. In dieser Gruppe entspricht die Destruktion der Mitochondrien morphologisch einer Veränderung in der Densität, mit elektronenmikroskopischen histochemischen Methoden können charakteristische Lokalisationen, Veränderungen in der Aktivität nachgewiesen werden.Die Veränderungen der 1, Gruppe können mit dem Lichtmikroskop nicht verfolgt werden, während die der 2. Gruppe sich auch auf diese Weise gut untersuchen lassen.Die Anfangsveränderungen legen die Vermutung nahe, daß sie Anzeichen der Verlaufsrichtung des Stromes sind.

Electron-cytochemical detection of endogenous nickel in the myocardium in acute carbon monoxide poisoning. Applicability of a new cytochemical technique in forensic medicine.

SummaryAfter acute carbon monoxide poisoning (inhalation, dogs; perfusion, rats; postmortem, humans) endogenous nickel was cytochemically detected in heart muscle by dimethylglioxime reaction. Dimethylglioxime cytochemical reaction, in each case at the COHb level, was always positive in experiments and in postmortem samples. A nickel ion accumulation in the heart muscle above 30 rel.% of COHb was suggested. A possible role of nickel ion in the pathomechanism of the acute carbon monoxide poisoning was supposed. This dimethylglioxime cytochemical technique is applicable in forensic medical practice, primarily because it is not disturbed by autolysis.ZusammenfassungBei unterschiedlicher Einwirkung von Kohlenmonoxid (Inhalation, Perfusion, post mortem) wurde endogenes Nickel im Herzmuskel histochemisch durch die Dimethyl-Glyoxim-Reaktion nachgewiesen. Die histochemische Reaktion ist für den morphologischen Nachweis ab 30% COHb geeignet. Die Methode ist für die Routinearbeit zu empfehlen, zumal die Reaktion nicht durch Autolyse gestört wird.

Use of neodymium nitrate in electron microscopic histochemical demonstration of ATP-ase

SummaryNeodymium nitrate has been used instead of lead nitrate in the modified Wachstein-Meisel incubation solution. Neodymium reacts easily with the anorganic phosphate groups. The rare earth metal has strong electron scattering property. It was used in 3,0 mM concentration in the incubation solution. The histochemical precipitate obtained, appeared to be selective and was well localizable. Neodymium nitrate seems to be suitable for the electron microscopic histochemical demonstration of hydrolytic enzymes.

Comparative topo-optical investigation of cardiac glycoside localization

SummaryThe aldehyde-bisulfite-toluidine blue (ABT) reaction is a selective topo-optical test of vicinal-OH and amino-OH groups. The localization of cardiac glycoside was investigated morphologically. After digoxin the sarcolemma membranes, capillaries and sinus node showed strong basophilia and negative birefringence. The topo-optical reaction are useful for the histopathological examination. The new method gives the possibility to the digoxin intoxication with a high specificity.ZusammenfassungDie Aldehyd-bisulfit-Toluidinblau (ABT) Reaktion wird in Geweben als selektive topo-optische Testmethode der vicinalen-OH-Gruppen und der Amino-OH-Gruppen angewandt. Die Lokalisation der herzwirksamen Glycoside wurde morphologisch untersucht. Nach experimenteller Digoxinintoxikation wurde das Reaktionsprodukt in den Sarcolemma-Membranen, den Sinus-Knoten und an den Kapillaren lokalisiert. Für die topo-optische Reaktion ist die durch die orientierte Farbstoffbindung herbeigeführte starke Doppelbrechung kennzeichnend. Die Unterschiede in der Doppelbrechung bieten eine Möglichkeit zur neuartigen Analyse in der Histopathologie bei Glycosidvergiftung.

A new method for cytochemical demonstration of calcium in heart muscle

SummaryThe ortho-cresolphtalein complex was successfully adapted for the electron microscopic cytochemical demonstration of calcium. The reaction product is of granular nature with sufficient electron density for finer localization. Intense precipitation was found on the sarcolemma and transverse tubules and in the sarcoplasmic reticulum. Myofilaments, mitochondria and capillary endothelial cells also showed a positive reaction. The electron microprobe analysis of the precipitate proved the presence of calcium. Disturbing effects of magnesium ions were prevented by the incorporation of 8-hydroxyquinoline in the incubation medium.