Unusual lamellar calcifications in two rare cases of splenic aneurysms associated with fibromuscular dysplasia.

Fibromuscular dysplasia (FMD) of the splenic artery is a rare underdiagnosed condition. Here, we report two cases of FMD affecting the splenic artery: one alone and one concomitantly with the renal artery. Histology revealed fibromuscular thickening of the media layer alternating with a circumferential calcification of the whole artery thickness. Ultrastructurally, FMD showed matrix vesicles and dense bodies in the extracellular matrix. A diagnosis of FMD with calcification was made. This is the first report to document circumferential lamellar calcifications alternating with the more typical fibrotic medial areas in the rare FMD localized to splenic artery.

[Age factors in architectonics of the splenic arterial vessels]. / Vozrastnye osobennosti arkhitektoniki arterial'nykh sosudov selezenki.

Spleen of 86 people of different ages was examined using the roentgenoanatomical, macromicroscopical, histological, morphometric and variation-statistical methods. It was established that, starting from the first mature age there was a steady reduction of the area occupied by arterial vessels, which reached its minimal value in senile age and in long-livers. In senescent persons, the atrophy of white and red pulp was established with a concomitant increase in spleen stromal elements content. The changes in the ratios of spleen tissue elements define the dynamics of age-related changes of its functions (including the immune function). In elderly and senile age these changes seem to be related with the reduction of adaptation capacities of the intraorgan arterial bed of the spleen.

Pathological morphology alteration of the splanchnic vascular wall in portal hypertensive patients.

OBJECTIVE: To investigate the pathological morphology alteration of the splanchnic vascular wall in portal hypertensive patients. METHODS: Splenic arteries, veins and gastric coronary veins from portal hypertensive patients (n = 50) were removed during esophagogastric devascularization with splenectomy and were observed under optic and electron microscopes. The expression of iNOS in the splenic artery wall was analysed with immunohistochemistry. RESULTS: The internal elastic membrane and medial elastic fibers of the splenic artery wall were broken and degenerated. Atrophy, apoptosis and phenotypic changes were seen in smooth muscle cells of splenic arteries. Positive staining for iNOS was seen in the cytoplasm of smooth muscle cells and iNOS activity was elevated compared with the non-cirrhotic patients (P < 0.01). In the splenic and gastric coronary veins of cirrhotic patients, we found proliferative intima, extensive thrombi adhering to the venous wall, mimicked arteriosclerosis plaques accompanied with hypertrophy of smooth muscle cells, and thickened muscle fibers of veins with increase in extracellular matrix. CONCLUSION: Portal hypertension may be complicated by splanchnic arterial and venous vasculopathy. There may be an interactive relationship among portal hypertension, splanchnic hyperdynamic disturbances and splanchnic vasculopathy in the pathogenesis of portal hypertension.

Electron microscopic cytochemical studies of anionic sites in the rat spleen.

The distribution patterns of the intensity of negative charge on the free surfaces (glycocalyx of the plasma membrane) of endothelial cells (ECs) in blood vessels and reticular cells (RCs) in the splenic cord of the rat spleen were studied by an electron microscopic cytochemical method using polyethyleneimine (PEI) as a cationic probe. Spleens from adult male rats were perfusion-fixed with 0.5% glutaraldehyde -4% paraformaldehyde containing 0.05% cetylpyridinium chloride and then perfused with 0.5% PEI at pH 7.4. On the free surfaces (glycocalyx of the plasma membrane) of the ECs examined, distinct PEI-positive reactions were observed in blood vessels, such as trabecular arteries, central arteries, arterial capillaries, pulp veins and trabecular veins. These PEI-positive electron-dense substances in the trabecular arteries, central arteries, and trabecular veins took the shape of a band of 170-250 nm in thickness. On the other hand, the corresponding ultrastructure of the ECs lining the splenic sinuses and the RCs in the splenic cord showed exceedingly weak PEI reactions. The PEI-reactive deposits were significantly thinner than those in the above blood vessels. As the thickness of the electron-dense substances can be related to the density of the negative charge, these results suggest that there is a high intensity of negative charge on the free surfaces (glycocalyx of the plasma membrane) of ECs in blood vessels where blood cells and plasma pass into the red pulp or are discharged from the red pulp. In contrast, the splenic sinuses and RCs, which are the main components of the red pulp, contain weakly negative-charged sites. This may contribute to the microcirculation of the splenic blood vessels and elucidate the possible physiological functions of the spleen, such as blood storage.

Histochemical analysis of acidic glycoconjugates in the endothelium lining the splenic blood vessels in the rat.

An electron microscopic analysis with specific histochemical stainings for acidic glycoconjugates was carried out to examine the endothelium lining blood vessels of the rat spleen. Histochemical staining performed was the postembedding high or low iron diaminethiocarbohydrazide-silver protein-physical development (HID or LID-TCH-SP-PD) method, with or without prior digestion with acidic glycoconjugate-degrading enzymes, such as heparitinase, testicular hyaluronidase, chondroitinase B and neuraminidase. The results indicated that the acidic glycoconjugates in the basal lamina of the endothelial cells lining the four types of blood vessel (central arteries, arterial capillaries, splenic sinuses and pulp veins) were heparan sulfate, chondroitin sulfate A and/or C, chondroitin sulfate B and sialic acid residues. In the endothelial cells lining the central arteries, arterial capillaries and pulp veins, the surface coat of the luminal plasma membrane included heparan sulfate, chondroitin sulfate A and/or C, chondroitin sulfate B and sialic acid residues, whereas the corresponding ultrastructure of the splenic sinuses was devoid of detectable amounts of acidic glycoconjugates. This suggests that such characteristic histochemical features of the endothelium in the four types of the splenic blood vessel can be related to the possible physiological functions of the spleen.

Indicial functions of arterial remodeling in response to locally altered blood pressure.

We investigated the effect of locally altered blood pressure on the remodeling processes of the cells and extracellular matrices of the splenic and ileal arteries and used an indicial function approach to quantitatively analyze the relationship between the altered blood pressure and the remodeling processes. Blood pressure in these arteries was locally modulated by constricting the aorta at a location between the celiac and mesenteric bifurcations, resulting in a higher blood pressure at the splenic arteries then at the ileal arteries, After the pressure changes, the cross-sectional areas and the fractions of the cells and extracellular matrices of the splenic and ileal arteries were examined by electron microscopy at 2, 6, 10, 20, and 30 days. We found that both arteries remodeled, but the splenic arteries (higher blood pressure) remodeled more rapidly and to a larger degree than the ileal arteries (lower pressure compared with the splenic arteries) of the same animal. To verify whether an identical change in the blood pressure at the splenic and ileal arteries leads to the same remodeling process in these arteries, we created another model by constricting the aorta at a location between the mesenteric and renal bifurcations, resulting in hypertension of the same level at both splenic and ileal arteries. We found that the remodeling processes of the cells and matrices were almost identical in the arteries with similar changes in blood pressure. Thus we conclude that the remodeling processes of cells and matrices of the splenic and ileal arteries are dependent on the local blood pressure in aorta constriction-induced hypertension, and the indicial analysis is a useful approach in the description of the relationship between the blood pressure and the arterial remodeling processes.

The relationship between density of alpha-adrenoceptor binding sites and contractile responses in several porcine isolated blood vessels.

1. The aim of this study was to investigate constrictor alpha-adrenoceptors in three isolated blood vessels of the pig, the thoracic aorta (TA), the splenic artery (SA) and marginal ear vein (MEV) and then compare the functional response with the densities of alpha 1- and alpha 2-adrenoceptor binding sites in these and several other porcine vascular tissues, palmar common digital artery (PCDA), palmar lateral vein (PLV) and ear artery (EA). 2. Noradrenaline (NA), phenylephrine (PE) and UK14304 (all at 0.03-10 microM) elicited concentration-dependent contractions in the TA and MEV, with a rank order of potency of UK14304 > NA > PE. UK14304 produced maximal responses which were 58% (TA) and 65% (MEV) of that of NA. In the SA, UK14304 and PE produced maximal responses which were less than 10% and 50% of the NA-induced maximal response respectively, with an order of potency of NA > PE. In the SA, NA-induced contractions were competitively antagonized by prazosin (pA2 = 8.60 +/- 0.15). Further, rauwolscine (1-10 microM) antagonized NA-induced contractions with an apparent pKB of 6.09 +/- 0.11 (n = 6), indicating an action at alpha 1-adrenoceptors. The combination of the two antagonists at concentrations selective for alpha 1- (0.1 microM) and alpha 2-adrenoceptors (1 microM) had no greater effect than either antagonist alone. This suggests that the SA expresses only post-junctional alpha 1-adrenoceptors. 3. In the TA, prazosin produced non-parallel shifts in the NA-induced CRC and this was also observed with rauwolscine, where reductions in the maximal responses were also observed. In the MEV, prazosin was largely inactive in antagonizing NA-induced contractions. In both these vessels a combination of these two antagonists had a greater effect than either alone, indicating the presence of functional alpha 1- and alpha 2-adrenoceptors. The post-junctional alpha 2-adrenoceptors in all of these vessels were resistant to prazosin, suggesting the alpha 2-adrenoceptor to be of the alpha 2A/2D subtype. The expression of functional alpha 2-adrenoceptors was MEV > TA > PLV > PCDA > SA. 4. In radioligand binding studies using TA P2 pellet membranes, [3H]-prazosin and [3H]-RX821002 ([1,4-[6,7(n)-3H] benzodioxan-2-methoxy-2-yl)-2-imidazole) labelled different high affinity sites, and in competition studies using identical membranes corynanthine displaced [3H]-prazosin with 10 fold higher affinity than rauwolscine, indicating that [3H]-prazosin was selectively binding to alpha 1-adrenoceptor sites. Further, rauwolscine displaced [3H]-RX821002 with approximately 100 fold greater affinity compared to corynanthine, which is indicative of selective alpha2-adrenoceptor binding.5. Separation of the P2 pellet into plasma membrane and mitochondrial fractions was carried out using a differential sucrose density gradient. [3H]-prazosin and [3H]-RX821002 binding sites were found in both the plasma membrane and mitochondrial fractions.6. In saturation studies all tissues produced single site saturation curves with no difference in the Kd(range 0.13-0.20nM) of the alpha1-adrenoceptor sites for [3H]-prazosin. However, there was considerable variation in Bmax of alpha 1-adrenoceptor sites; the highest density was found in the TA (397.9 =/- 52.7 fmol mg-1, n = 4), followed by the PCDA (256.7 +/- 22.7 fmol mg-1, n = 4), the PLV and SA having approximately equal density (143.6 +/- 3.9 and 159.1 +/- 7.0 fmol mg-1 respectively, n = 4 for both), followed bythe EA (91.3 +/- 10.5 fmol mg-1, n = 3) and the MEV had the lowest density (48.9 +/- 11.4 fmol mg-1,n = 3).7. In saturation studies using [3H]-RX821002, all tissues produced single site saturation curves with no differences in the Kd values (range 1.31 +/- 2.16 nM) but the highest densities were found in the TA and MEV (545.3 +/- 36.2 and 531.0 +/- 40.9 fmol mg-1 respectively), followed by the PLV (418.4 +/- 39.4 fmol mg-1), then the EA (266.3 +/- 40.0 fmol mg-1), and low densities of [3H]-RX821002 binding being found in the PCDA and SA (155.9 +/- 18.1 and 117.5 +/- 19.3 fmol mg-1 respectively).8. The pattern of binding site distribution for alpha l- and alpha 2-adrenoceptors is in reasonable agreement with functional studies carried out in these porcine vascular tissues; the TA has the highest densities of alpha 1-and alpha2-adrenoceptors; in the SA and PCDA there is a predominance (although small) of alpha l-adrenoceptor binding sites, the reverse of which is observed both in the PLV and MEV (i.e. greater density of alpha2-adrenoceptor sites). Thus, it would appear that alpha 1- and alpha2-adrenoceptor densities play a role in the expression of functional responses via these receptor subtypes; although it is interesting to note that the SA did have a small density of alpha 2-adrenoceptor binding sites, no functional response was observed after alpha2-adrenoceptor activation.

Electron microscopic studies on reticular fibers in the pig sheathed artery and splenic cords.

The reticular fibers of the capillary sheath and splenic cord were studied in the pig spleen by transmission electron microscopy to reveal their components and the presence of sialic acid in the amorphous ground substance. Collagen fibrils, elastic fibers, microfibrils, nerve fibers and smooth muscle cells were observed in the reticular fibers of the splenic cord, while only microfibrils were recognized in the reticular fibers of the capillary sheath. The binding of LFA lectin to the splenic cord was stronger than the capillary sheath. These findings may suggest that the reticular fibers of the splenic cord include numerous functional elements and perform an important role during contraction or dilation of the spleen. On the other hand, it seems to be reasonable to term the capillary sheath as "capillary basement membranous reticular tissue", since the reticular fiber of the capillary sheath resembles the basement membrane of the capillary in its components.

"Intermediate zone" of mammalian spleens: light and electron microscopic study of three primitive mammalian species (platypus, shrew, and mole) with special reference to intrasplenic arteriovenous communication.

The intermediate zone (IZ) of nonperfused and perfused spleens in three species of primitive mammals (shrew, mole, platypus) was studied morphologically. The IZ is a tissue zone consisting of plexiform vessels, probably venous capillaries, and is located transitionally between the white and red pulp. The IZ is separated from the white pulp by the arterial net (AN), in which the white pulp arteries terminate. Development of the IZ differs between the three species examined being distinctive in the platypus and shrew. The IZ is thin in the mole spleen. A closed type of arteriovenous (A-V) anastomosis was demonstrated in or around the IZ in the two Insectivora species examined. In the shrew spleen, peripheral arterial branches running within the IZ anastomose with the AN around the follicle. The AN anastomoses eventually with venous plexiform vessels of the IZ around the nonfollicular area of the white pulp to form a closed system. In the mole spleen, A-V anastomoses were noted between white pulp arteries (follicular and AN) and veins of the red pulp, either by direct communication or through fenestrated IZ vessels compatible with the plexiform vessels of the shrew spleen. A-V anastomosis in the IZ is probable, but not confirmed, in the platypus spleen, as analysis was limited to a nonperfused specimen. Well-developed ellipsoids were noted around arterial terminals of the IZ in the shrew spleen. Ellipsoids were also noted around all arterial terminals of the mole spleen directed to the red pulp. Most ellipsoids of the mole spleen appeared located within the IZ. No ellipsoids were present around arterial terminals of the IZ in the platypus spleen. Closed circulation was noted in terminals of the pulp artery in spleens of all three species. All pulp arteries of the mole spleen are postellipsoid segments of white pulp (AN and follicle) arteries. No ellipsoids were found around terminals of the pulp artery (penicillar artery) in shrew and platypus spleens. The IZ is probably homologous to the perilymphatic sinusoid (vein) of the lungfish spleen and may be regarded as part of the red pulp. The IZ may be representative of primitive mammalian spleens that have closed circulation. The marginal zone (MZ) of common mammalian spleens is probably a modified IZ by differentiation (remodelling) of the intrasplenic vein. In this process, withdrawal of venous vessels from the IZ occurred, leaving a lymphoreticular zone with open circulation (MZ). The marginal sinus reported in some mammalian spleens is probably a modified AN formed during this process. Possible morphological alterations of the spleen in vertebrate phylogeny are discussed.

Fine architecture of the splenic terminal vascular bed as revealed by arterial and venous pressure-loading perfusion fixation.

The three-dimensional fine architecture of the red pulp of human and animal spleens, which as fixed by a modified version of the arterial and venous pressure-loading perfusion fixation (AVPL perfusion fixation) method, is demonstrated by scanning and transmission electron microscopy. In the human spleen, changes in splenomegalias associated with hereditary spherocytosis and chronic portal hypertension are also introduced in addition to the normal architecture of the red pulp of spleens removed from patients with stomach cancer. The AVPL perfusion fixation of these spleens clearly visualized complicated three-dimensional fine architecture of the red pulp and provided much important information on in situ morphology and dynamic change of the terminal vascular bed, including venous pressure-dependent size change of the stomata and three-dimensional shapes of the capillary terminal, with positive proof of their opening into the cordal reticular tissue. In studies of the spleen associated with portal hypertension, the AVPL perfusion fixation is considered a necessary technique for analysis of the structural deviation closely relating to a very high venous pressure.

Fibroblasts and sympathetic innervation of blood vessels.

The distances between fibroblasts and adrenergic varicosities were measured in four distinct blood vessels of the dog, concomitantly with the measurement of corresponding distances between smooth muscle cells and adrenergic varicosities. The results showed a closer anatomical relationship between fibroblasts and adrenergic varicosities as compared to that of adrenergic varicosities and smooth muscle cells. These morphological results indicate that sympathetic neurotransmitter(s) may attain at the fibroblast membrane concentrations similar to those occurring at the smooth muscle cell membrane.

Electron microscopic localization of factor-VIII-related antigen in adult human blood vessels.

We have localized factor-VIII-related antigen, using immunofluorescence and electron microscopy, in adult human blood vessels. In addition to its presence in endothelial cells, the antigen was localized within subendothelium and the layers of elastic lamina closest to the lumen. Also, we provide the first morphological evidence that factor-VIII-related antigen is associated with collagen fibrils within the vessel wall. These studies suggest that this subendothelial factor-VIII-related antigen may play a role in the adhesion of platelets to subendothelial components following endothelial injury.

Morphological studies of the spleen in idiopathic portal hypertension (so-called Banti's syndrome without liver cirrhosis) using light microscopy, scanning electron microscopy and histometry.

Morphological changes in the spleens of patients with idiopathic portal hypertension (IPH) were studied and compared with the normal spleen. The study used (1) light microscopy with histometry, (2) scanning electron microscopy (SEM) of the splenic tissue with histometry and (3) SEM of the spleen vascular replica. Histometrical studies by light microscopy showed that the volume of red pulp of IPH was increased in a unit area and to a total of 12 times the normal in the whole spleen. The white pulp was scanty of lymphocytes and decreased in a unit area but it was increased in the whole spleen. SEM of the white pulp of IPH demonstrated many channels formed by reticulum cells and running parallel with each other along the central artery. This finding presumably corresponds to periarterial fibrosis in light microscopy. SEM histometry demonstrated that the venous sinuses of IPH were small but increased in number and occupied the same percentage area in a unit red pulp area as in the normal spleen. The Billroth cord of IPH was narrowed and occupied by thickened reticulum cells, which may cause increased pooling and destruction of blood cells in the enlarged spleen (hypersplenism). SEM of the tissue and vascular replica demonstrated open arterial termination in the Billroth cord in the spleen of IPH as well as in the normal spleen. Venous sinuses in the replica of IPH ran parallel with each other forming bundles with fewer intercommunications than normal.

Studies of the hemopoietic microenvironment. VIII. Andrenergic and cholinergic innervation of the murine spleen.

Neurohistochemical techniques were used to confirm morphologically the distribution of adrenergic and cholinergic nerves to the splenic microvasculature. The results form the basis of this report. Using these methods, adrenergic innervation was observed only in the adventitia of arteries and arterioles. No cholinergic innervation was found in this site. No adrenergic or cholinergic innervation could be demonstrated to the channels of the red pulp, venules or veins. These data provided morphological evidence that in the murine spleen only splenic arteries and arterioles are innervated; and these have only an adrenergic innervation.

A morphologically different type of smooth muscle cell in the inner media of the splenic artery.

A morphologically different cell type to the typically smooth muscle cell has been found in the inner medial layers of the splenic artery. Besides the morphological differences, typical criteria for a smooth muscle cell are demonstrated, giving an account of their identity. Morphological details are described. The importance of these cells as regards lesions of the vessel wall is briefly discussed.