Kimmo Kyösola

Cold Injury of the Rat Skin: A Fluorescence Histochemical Study of Adrenergic Nerves, Mast Cells and Patency of Cutaneous Blood Vessels

In order to investigate the pathogenesis of frostbite a combined trypan blue and catecholamine fluorescence method was adopted for studying changes in cutaneous adrenergic nerves, mast cells, the patency of blood vessels, the distribution of blood flow and vascular permeability after experimental cold injury. The skin of rats was frozen with a copper cylinder precooled in liquid nitrogen. The rats were investigated 1/2, 4, 8, 16 and 24 h or 2, 4, 7, 9 and 14 days after freezing. Trypan blue has a red fluorescence when absorbed onto proteins. In normal skin this stain was seen inside the blood vessels and in the dermal tissues. Bluegreen fluorescent adrenergic nerves were seen in the erector pili muscles and around arteries and arterioles. The fluorescence of the mast cells was yellow. In the specimens taken half an hour after skin freezing most of the adrenergic nerves appeared morphologically normal. Degenerative changes were observed in the mast cells varying from degranulation and diminution of yellow ...

Reinnervation of Human Skin Grafts: A Histochemical Study

The reinnervation of nine human skin grafts was investigated using histochemical thiocholine methods for the demonstration of cholinesterases. The regenerated cutaneous nerves showed both specific acetylcholinesterase and nonspecific cholinesterase reactions. In the youngest specimens, taken 3 weeks after the grafting, such regenerated nerves were seen both at the subdermal level under the graft and at the margins of the graft. These nerves seemed to orient toward the denervated graft area. The growing nerves were generally distributed in a random fashion. The reinnervation of some hair follicles, erector pili muscles, and sweat glands were observed in well-innervated full-thickness and thick partial-thickness skin grafts. It is suggested that this target-organ control of regenerating nerves occurs as a result of the action of chemotactic factors. A well-innervated graft bed seems to be important for optimum reinnervation of skin grafts. Fibrosis and scarring seem to hamper nerve regeneration.

Fluorescence histochemical and electron-microscopical observations on the innervation of the atrial myocardium of the adult human heart

The existence of both adrenergic and cholinergic innervation of the atrial myocardium of the adult human heart was demonstrated by means of fluorescence induced by formaldehyde or glyoxylic acid and by electron microscopy. The adrenergic fluorescing axons (1) followed the course of blood vessels as typical perivascular nerve plexuses, and (2) formed a three-dimensional fairly dense nerve net obviously not related to the blood vessels. The varicosities frequently came into close apposition on myocardial cells. Several types of nerve terminals were differentiated at electron microscopy: (1) an “adrenergic” type containing small (diameter 450–700 Å) dense-cored vesicles and usually (in various proportions) small “empty” and/or large (900–1500 Å) dense-cored vesicles, (2) a “cholinergic” type containing small (ca. 500 Å) “empty” vesicles and occasionally also some large (mean diameter ca. 1200 Å) dense-cored vesicles, (3) a “pale” type containing only a few or no vesicles, (4) a “disintegrated” type containing degenerated mitochondria, autophagic vacuoles, and occasional normal-looking mitochondria, (5) nerve terminals containing a large number of mitochondria in addition to varying vesicle populations, and (6) a (possibly baroreceptive type of) nerve terminal containing myelinlike lamellated structures. The “disintegrated” and the “pale” types of nerve terminals possibly represent different stages of axonal degeneration, or may correspond to diminution in the transmitter substance concentration under certain pathophysiologic conditions, respectively. Nerve terminals crowded with mitochondria may be sensory and involved in mechano-or chemoreceptive functions. In preliminary experiments convincing evidence was obtained that the glyoxylic acid-induced fluorescence histochemical method will be suitable for comparative studies on (human) clinical specimens, e.g., for analyzing the degree of the functional activity of the intrinsic adrenergic innervation of the myocardium under various pathophysiologic conditions. The modification which appeared most appropriate for such studies is described in detail, and is proposed for use as a standard method in other similar or related studies on human clinical series. The essential criteria for analyzing the specimens at fluorescence microscopy are suggested as well.

Studies of auricular catecholamines by fluorescence histochemistry in various heart diseases of man

A comparative histochemical and clinical study concerning the state of the intrinsic adrenergic innervation of the human atrial myocardium was carried out, using the glyoxylic acid-induced fluorescence histochemical method. Specimens from the right auricular appendage were obtained during open-heart surgery from patients suffering from 1. ischaemic heart disease (IHD), 2. atrial septal defect of the secundum type (ASD), and 3. left-sided univalvular or multivalvular heart disease (VHD) with or without congestive heart failure (CHF) experienced prior to surgery. In the IHD group the densities of both the perivascular and the “free” myocardial adrenergic nerve net were greater than in the ASD group and especially in the VHD/ CHF group. Secondly, the intensity of fluorescence of the adrenergic structures was generally higher in the IHD group than that in the VHD/CHF group. Further, the average size of the varicosities, the number of varicosities per given length of axon, and the proportional share of the large varicosities were greater in the IHD group than in the ASD and VHD/CHF groups. The difference between the IHD and ASD groups was not great but was obvious in any case. In some patients with VHD/CHF fluorescing axons were observed only occasionally, and the tiny varicosities exhibited a hardly discernible fluorescence. Thus the amount of noradrenaline (NA) in the adrenergic fibres in the IHD group seems to be higher than in the ASD and especially VHD/CHF groups. The high level of NA in the IHD group is assumed to constitute a contributory factor in both intracellular metabolic changes and the systemic changes typical of myocardial ischaemia and infarction. In one patient with IHD and in six patients with VHD/CHF with significantly higher heart volume (mean±SD) compared with the rest of the patients (P<0.001), huge local axonal accumulations of NA in the form of “droplet fibres” were found. These enlarged, bulging adrenergic axons are assumed to be a consequence of mechanical trauma with stretching or disruption of the axons due to myodegenerative processes. It is further assumed that these “droplet fibres” are relatively common in those patients with diseased myocardium. They may constitute an extra contributory factor to the tendency to arrhythmias so typical of patients of this kind, by increasing the excitability of non-automatic tissue.

Time course of β2-integrin cd11b/cd18 (mac-1, αmβ2) upregulation on neutrophils and monocytes after coronary artery bypass grafting: CD11b upregulation after CABG surgery

Although upregulation of CD11b/CD18 receptor, i.e. activation of neutrophils and monocytes, during cardiopulmonary bypass is well documented, the duration of the active state after uncomplicated operation is less understood. We therefore investigated CD11b expression of phagocytes in blood samples collected 2-4, 24, 48 and 72 h after coronary artery bypass grafting. CD11b expression on neutrophils was significantly elevated at 2-4 and 24 hours after operation as compared with baseline. On monocytes, expression peaked at 24 h and returned to baseline by 72 h. Because CD11b is a sensitive marker, effects of different sampling techniques on its expression were also studied. CD11b expression was similar in samples collected with a syringe from arterial or central venous catheter or with open technique from cubital vein. On neutrophils from healthy subjects, sampling with syringe caused small (10%) but statistically significant increase of expression. We conclude that activated neutrophils disappear from circu...

Plasma cell granuloma of lung and pleura

A report is presented of seven patients with plasma cell granuloma of the lung or pleura. Two were operated on in 1970, three in 1971, one in 1973 and one in 1984. None of the patients had smoked cigarettes. Five had a history of infection. Most of the patients were young, healthy and working. The surgical treatment was lobectomy in four cases, bilobectomy in one case and resection of tumour in two cases. The histology of the tumours was re-evaluated, and was similar in all cases. They were composed mainly of plasma cells, lymphocytes and granulocytes, and small foci of calcification were also seen. No recurrence of tumour was found in observation up to 13 years postoperatively. One patient died of pancreatic carcinoma after 6 years. The others are doing well.

Adrenergic innervation of the human gall bladder

SummaryAdrenergic innervation of the human gall bladder was studied using two specific fluorescence histochemical methods. Blue-green fluorescing varicose nerves were scarce and mostly followed the course of blood vessels as typical perivascular plexuses. However, some adrenergic nerves not associated with the vessels were occasionally seen, as well as structures suggestive of a pericellular arrangement of varicose adrenergic nerve terminals on non-fluorescing ganglion cells. A few enterochromaffin cells were seen in the epithelial lining, also in the deep invaginations obviously representing the Aschoff-Rokitansky sinuses. Occasionally, small rounded cells with a rounded, relatively large nucleus, and exhibiting a weak yellow-green to blue-green granular cytoplasmic fluorescence, were observed in the wall of the gall bladder. The possible functional and evolutionary significance of these neural and endocrine elements was discussed against the data on physiological and pharmacological studies obtained from the literature. It was concluded that their significance is, in all probability, secondary to the influence of the intestinal polypeptide hormones, vagal innervation and circulating catecholamines upon the normal function of the gall bladder. The glyoxylic acid-induced fluorescence histochemical method was found to be superior to the conventional formaldehyde technique in studies on human tissue.

Adrenergic innervation of the choledocho-duodenal junction of the cat and the dog

SummaryThe adrenergic innervation of the choledocho-duodenal junctions of cats and dogs was studied using the formaldehyde-induced fluorescence method for histochemical demonstration of catecholamines.A fluorescent network of nerve fibres was demonstrated in the fibromuscular layer suggesting direct innervation of the sphincter muscles. “Basket-like” fluorescent structures suggestive of pericellular networks around non-adrenergic ganglia were seen in the plexus of Auerbach, on the surface of the sphincter muscles and inside the muscles. Fluorescent nerve cells were observed in the plexus of Auerbach, sending their fibres towards the ductus choledochus. Small cells exhibiting a very intense yellow fluorescence were seen slightly outside the smooth sphincter muscle. Another type of intensely yellow fluorescent cells was seen around blood vessels inside the sphincter muscles and in the lamina propria of the ductus choledochus.It is concluded that the adrenergic innervation of the sphincters regulating the flow of bile in cat and dog involves three different mechanisms:1)direct innervation of the muscle,2)adrenergic innervation of the intramural non-adrenergic ganglia,3)influence of the intramural fluorescent,catecholamine-containing cells of three different types upon the smooth muscle either directly or indirectly.

The adrenergic innervation of arteriovenous anastomoses in the subcutaneous fascia of rat skin.

Stretch preparations of the subcutaneous fascia of rat trunk skin were studied using the glyoxylic acid-induced fluorescence technique for demonstrating adrenergic nerves. To facilitate the identification of blood vessels some rats were, in addition, injected intravenously with a fluorescent (FITC-dextran) tracer and some stretch preparations were subsequently stained with haematoxylin and eosin. The arteriovenous anastomoses were found to be innervated with fluorescent adrenergic nerves. Several types of arteriovenous anastomoses were identified in the subcutaneous fascia of rat trunk skin: 1) Several quite densely innervated arteriolar branches arising from the segmental subcutaneous artery and terminating in the accompanying vein and in the capillary bed. 2) Simple arteriovenous connections, with a moderately dense adrenergic innervation usually restricted to the beginning of the arteriole. 3) More specialized arteriovenous anastomoses which often showed a slightly tortuous or S-shaped course. They oft...

Cholinesterases of the gall bladder

SummaryCholinesterase histochemistry of the human gall bladder was studied using two specific methods.Distribution of acetylcholinesterase: In the mucosa, nerve fascicles consisting of densely packed parallel single nerve fibres, small ganglia and “spot”-and “glomerule”-like concentrations of acetylcholinesterase activity were observed. In the muscle layer, a wide-meshed network of delicate nerves, with occasional areas of very dense innervation, and small ganglia were seen. In the serosa, “glomerule”-like structures surrounded by dense “baskets” of delicate nerves were observed. — The general scheme of distribution of non-specific cholinesterases was similar to that of acetylcholinesterase.It seems that the cholinergic innervation of the gall bladder is related to both secretion and absorption and contractility. Some cholinergic nerves are probably sensory, especially because acetylcholinesterase-positive structures, possibly pressure of stretch receptors, supplied with nerves were observed in the mucosa and the serosa. The cholinergic innervation of the gall bladder muscle was scarce except occasional areas of very dense innervation. It may thus be concluded that the intermuscle spread of excitation plays an important role, the majority of the smooth muscle cells receiving their nervous influence via electrotonic coupling.