Mechanisms of neovasculogenesis and its regulation in the adult organism

Under regeneration of organs, wound healing, tumour growth, inflammatory processes, under many compensatory and adaptive reactions in the organism of mature persons and animals, an inevitable formation of new blood vessels (neovasculogenesis) takes place. Modern notions on mechanisms of neovasculogenesis are based on the fact that new formation of vessels in a mature organism includes processes of migration and replication of endothelial cells according to the principle: "endothelium from endothelium". The literature data on neovasculogenesis in the mature organism are summarized and compared with the authors' investigations. Characterization of new blood vessels growth is presented; ultrastructural organization of endotheliocytes in growing capillaries, formation of barrier-transport properties in the newly formed vessels, role of inductors and inhibitors of neovasculogenesis in creation of new vascular formations are considered.     

Mitotic activity of pial microvascular endothelium rat in postnatal ontogenesis

Mitotic activity of endothelium in pial microvessels has been studied with the aid of intra-vessel autoradiography in 4-, 12- and 30-day old rats using these parameters: number of labeled nuclei per vessel or per 100 mcm of its length, or per 1 mm2 endothelial surface area, as well as number of vessels with labeled endothelium cells. The first parameter was independent of vessel diameter. The other parameters had highest value in the pericapillary vessels in all rats. These values decrease with rat age. The highest rate of brain growth was revealed after reaching the greatest mitotic endothelium activity in pial bed.     

Age and changes in the number of endocrine cells in the stomach and their role in senile atrophy of the gastric glands

In the gastric mucous membrane of 40 rats representing 4 age groups (pubertal, mature, old and very old age) per 10 animals in each, thickness of the mucous membrane has been measured by the argyrophil method of Grimelius and the argentaffin method of Masson - Hamperl and the number of the endocrine cells has been counted. The thickness of the mucous membrane and the length of its glands, as well as the number of argyrophil cells increase from the pubertal towards the mature age. From the mature up to the old age the number of argyrophil cells rises in the antral part, and in the acid-producing zone it significantly decreases. The number of argentaffin cells in the stomach does not change from the pubertal towards the old age. In the very old age, against the background of atrophy of gastric glands, the number of both argyrophil and argentaffin cells decreases in both parts of the stomach studied. The data obtained are discussed taking into consideration functional role of various types of endocrine cells, producing biologically active peptides and monoamines.     

Developing testicular microvasculature in the golden hamster, Mesocricetus auratus: a model for angiogenesis under physiological conditions.

The ultrastructure of the developing testicular microvasculature in the testes of immature (3, 5, 8, 10, 12, 16, 20, 25, 30 and 35 days old) golden hamsters was examined and compared to the testicular microvasculature of adult (3 months old) hamsters. In addition, in 16- to 35-day-old hamsters vascular permeability was studied after localization of injected horseradish peroxidase (HRP). Angiogenic processes were present in the testes of all examined immature hamsters and were most conspicuous between 8 and 25 days of age. These processes were absent in the testes of 3-month-old hamsters. On days 3 and 5, few undifferentiated blood vessels with activated endothelium were present in the interstitial spaces. Endothelial cell migration started from these 'mother vessels' and led to invasion of intertubular spaces by vascular sprouts, before vascularization of peritubular spaces occurred (after day 12). Sprouting endothelial cells were identified by the presence of a basal lamina and characterized by abundant cytoplasm and cell organelles. HRP-positive slits were seen in developing vessels, which opened to form the vascular lumen. HRP exited the vascular lumen through unspecialized endothelial contacts and micropinocytotic vesicles. By day 16, the blood-testis barrier prevented HRP from entering the seminiferous tubules beyond the basal compartment. By days 30 and 35 most testicular microvessels and at the age of 3 months all testicular microvessels were of the mature type, with narrow inactive endothelium and specialized cell contacts (including tight junctions). These results demonstrate that the postnatal vascularization of the testis in the golden hamster is a timed complex process. Due to high permeability, vascular sprouts are likely to influence the metabolic situation and thus the maturation processes of the testis. Angiogenesis in the golden hamster testis shares typical morphological features with angiogenic processes in other organs and species under various pathological and physiological conditions. We therefore conclude that the postnatal testis can be viewed as a physiological model of angiogenesis.     

Bird community composition patterns in urban parks of Madrid: The role of age, size and isolation

This study intends to assess the influence of fragment age, size and isolation (from the regional species pool) on bird community composition patterns in urban parks in Madrid, and the role of local and regional factors on community structure. Park age was a good indicator of habitat complexity. Park age and area accounted for 62% of the variability in species richness, but two measures of isolation from the regional species pool were not included as significant factors. Species composition in urban parks showed a high degree of nestedness, which was associated with park age and area, but not with two measures of isolation from the regional species pool. The degree of nestedness increased with park age; the distribution of species varying from nested in old and mature parks to random in young parks. The incidence (% of species occurrence in parks) in young parks was correlated with regional densities, whereas in mature and old parks the incidence was correlated with local densities. In this urban landscape, species composition appears to be regulated by local factors (particularly in mature and old parks), such that species accumulate in an orderly (not random) fashion in relation to park age and area. Regional influences seem to be more pronounced only in young parks, which are mainly colonized by species from the regional species pool.     

Expression of endothelial nitric oxide synthase in the vascular wall during arteriogenesis

Nitric oxide (NO) has been demonstrated to play an important role in angiogenesis, and also to be involved in collateral vessel growth. The expression of endothelial NO synthase (eNOS) is moderated partly by blood flow-induced mechanical factors, i.e., shear stress. The purpose of this study was to evaluate how the expression of eNOS correlates with the development of collateral vessels in dog heart, induced by chronic occlusion of the left circumflex artery. Immunoconfocal microscopy using an antibody against eNOS was used to detect expression of eNOS in different stages of arteriogenesis. Collateral vessels were classified into normal, growing and mature vessels by using the cytoskeleton marker desmin. Expression of the growth factors bFGF and metallproteinase-2 (MMP-2) was also examined. The data show that in normal arteriolar vessels, expression of eNOS is very low, but in growing collateral vessel there is a 6.2-fold increase, which, however, returned to normal levels in mature collateral vessels. The expression of eNOS was localized only in endothelium, either in normal or growing vessels. bFGF was very weakly stained in normal vessels, but highly expressed in growing collateral vessels. MMP-2 was strongly stained in neointima, but very weak in endothelium. In addition, we also examined expression of iNOS because iNOS may be induced in vessel injury or in disease states, but it was not detected in either normal or growing collateral vessels. Our findings indicate that the expression pattern of eNOS is closely associated with the development of collateral vessels, suggesting that eNOS plays an important role in arteriogenesis.     

Expression of endothelial nitric oxide synthase in the vascular wall during arteriogenesis

Nitric oxide (NO) has been demonstrated to play an important role in angiogenesis, and also to be involved in collateral vessel growth. The expression of endothelial NO synthase (eNOS) is moderated partly by blood flow-induced mechanical factors, i.e., shear stress. The purpose of this study was to evaluate how the expression of eNOS correlates with the development of collateral vessels in dog heart, induced by chronic occlusion of the left circumflex artery. Immunoconfocal microscopy using an antibody against eNOS was used to detect expression of eNOS in different stages of arteriogenesis. Collateral vessels were classified into normal, growing and mature vessels by using the cytoskeleton marker desmin. Expression of the growth factors bFGF and metallproteinase-2 (MMP-2) was also examined. The data show that in normal arteriolar vessels, expression of eNOS is very low, but in growing collateral vessel there is a 6.2-fold increase, which, however, returned to normal levels in mature collateral vessels. The expression of eNOS was localized only in endothelium, either in normal or growing vessels. bFGF was very weakly stained in normal vessels, but highly expressed in growing collateral vessels. MMP-2 was strongly stained in neointima, but very weak in endothelium. In addition, we also examined expression of iNOS because iNOS may be induced in vessel injury or in disease states, but it was not detected in either normal or growing collateral vessels. Our findings indicate that the expression pattern of eNOS is closely associated with the development of collateral vessels, suggesting that eNOS plays an important role in arteriogenesis. (Mol Cell Biochem 264: 193–200, 2004)     

Variability in the myeloarchitectonics of the nerves of the thyroid arteries during postnatal human development

Serial transversal sections on the nerve-vascular complexes of four thyroid arteries in newborns, in persons of the 1st and 2d periods of mature age, in elderly and old persons (40 complexes in each age group) are stained after Weigert--Pahl. Amount of myelin fibers (MF) of three dimensional classes are defined micromorphometrically. Total amount of the MF in the nerves of all thyroid plexuses is the greatest in the persons belonging to the second mature period of age: it is 2.7 times as great as in the newborns. In the elderly persons it is 1.5 times, and in the old persons--1.8 times as little as in the persons of the second period of maturity. Thin MF make the absolute majority. The amount of the MF with middle caliber in the thyroid nerves of the mature persons is 6.6 times as great as in the newborns. Their amount in the old persons is 1.7 times as little as in the mature persons. Thick MF are absent in the newborns; in the mature and elderly persons they are observed now and then. The thick MF are characterized by a high variation coefficient (up to 84%). According to the criteria of absolute and relative contents of both total and group composition of the MF in postnatal period, three stages in development of the thyroid innervational connections performed by the MF are revealed: productive, stabilization and involution. The correlative analysis reveales a direct dependence in the system diameter of each thyroid artery--number of the MF in nerves of its plexus in the newborns and mature persons. This dependence becoms negative in the elderly and old persons.(ABSTRACT TRUNCATED AT 250 WORDS)     

喉癌组织中类高内皮微静脉形态学及蛋白多糖表达的研究

目的:通过对喉癌组织中类高内皮微静脉的分布、超微结构和蛋白多糖表达的观察,探讨淋巴细胞归巢的抗肿瘤意义及蛋白多糖对淋巴细胞归巢的调节作用。方法:应用透射电镜及阳性胶体铁染色方法,观察和研究32例喉癌组织中的类高内皮微静脉的分布、超微结构和蛋白多糖的表达。结果:(1)早期病变肿瘤周边组织内可见大量类高内皮微静脉,其内皮细胞高大,胞浆突起增多,细胞核大,细胞器丰富,有大量淋巴细胞穿越管壁,可见大量肿瘤浸润淋巴细胞存在;晚期病变组织内类高内皮微静脉少见,淋巴细胞穿越管壁不活跃,肿瘤浸润淋巴细胞明显减少。(2)类高内皮微静脉壁,特别是邻近管腔侧与阳性胶体铁呈强阳性反应,在有淋巴细胞穿越的部位反应更加明显,而内皮扁平的血管基本不反应或反应微弱。结论:(1)喉癌组织中部分毛细血管后微静脉可演变为类高内皮微静脉,是淋巴细胞向癌组织中浸润(淋巴细胞归巢)的重要场所;(2)肿瘤组织中的淋巴细胞归巢与抗肿瘤密切相关;(3)蛋白多糖于淋巴细胞归巢旺盛的类高内皮微静脉呈强表达,可能对归巢淋巴细胞穿越类高内皮微静脉有调节作用。     

A quantitative study of changes in the human corpus luteum microvasculature during the menstrual cycle

Endothelial cells are the most abundant cell type in the corpus luteum (CL), and changes in blood vessels have been proposed to play a pivotal role in CL regression. We have studied quantitatively the changes in the human granulosa-luteal microvasculature in CL of various ages: young (Days 17-19 of the cycle), mature (Days 20-24), old (Days 25-27), early regressing (follicular phase of the following cycle), and late regressing (luteal phase of the following cycle). Blood vessels were identified by immunohistochemical staining for the endothelial cell marker CD34. Because of the anisotropy of blood vessels, both vertical and transverse sections of the granulosa-lutein layer (GLL) were used to estimate relative (volume, surface, and length densities) and absolute (mean cross-sectional area) vascular variables. Full luteinization from young to mature CL was accompanied by a 61% increase in the mean cross-sectional area of vascular profiles and a 52% increase in the mean volume of granulosa-lutein cells, as an estimator of changes in the volume of the GLL. In old and early regressing CL, there was a progressive increase in relative structural vascular variables, due to the shrinkage of the GLL, whereas the mean cross-sectional area of capillaries showed a 53% decrease from mature to old CL. Finally, in late regressing CL, there was a decrease in most relative structural variables, in spite of the increasingly shrunken GLL. The decrease in the capillary diameter found at the late luteal phase most likely leads to a decreased blood flow, and early changes in blood vessels could initiate and/or accelerate CL regression.     

Immunoelectron microscopic study of PECAM-1 expression on lymphatic endothelium of the human tongue

The expression of platelet-endothelial cell adhesion molecule-1 (PECAM-1) on lymphatic and blood vessels of the human tongue was examined with fluorescence and transmission electron microscopy (TEM). The study used anti-desmoplakins antiserum for light microscopic identification of the lymphatic vessels, plus a pre-embedding immunogold electron microscopic technique for TEM observations. Before making TEM observations, cryostat serial sections were immunostained with anti-desmoplakins or anti-PECAM-1 and then embedded. Semithin sections from each cryostat section were photographed under a light microscope and compared in order to identify the lymphatic vessels expressing PECAM-1. In fluorescence microscopy, PECAM-1 expression on lymphatic vessels was weaker than that on blood vessels. TEM observations showed that PECAM-1 expression on the blood vessels was observed only on the luminal surface of the endothelium. In lymphatic vessels, PECAM-1 expression was found both on the luminal and abluminal surfaces of the endothelium. The density of the PECAM-1 reaction products was lower in lymphatic vessels than in blood vessels. The density of PECAM-1 reaction products on the luminal surface of lymphatic vessels was higher than on the abluminal surfaces. The results suggest that blood vessels are more active than lymphatic vessels in leukocyte migration. The expression of PECAM-1 on the abluminal surface of lymphatic endothelium may allow leukocytes to adhere to the endothelium and interact in their migration from tissue into lymphatic vessels.     

Endocardial surface and atrial morphological changes during development and aging

Light, scanning, and transmission electron microscopic observations related to morphological changes of the right atrium as well as the atrial endocardium during development (15th embryonic day and 1 day old) and aging (560 days old) in the Syrian hamster were described and correlated. From the fetus to the adult, the atrial endocardium differentiates in parallel with, or in response to, the subjacent proliferating myocytes in the atrial wall and the trabeculae. Simultaneously, the atrium compartmentalizes grossly into a main chamber and an appendicular region. There is a progressive differentiation from a rudimentary, open chamber with primitive mural ridges in the fetal atria to a distinct, separate, atrial main chamber and appendage with a dense network of trabeculae in the adult. The fetal and neonatal endocardial, endothelial cells are convex with a central nuclear bulging and attenuated cytoplasmic extensions; the adult endocardium shows a squamous endothelium. Two cell surface specializations were observed in all age groups: microvilli and blebs or cytoplasmic protrusions. The general atrial morphology and surface endocardial changes were correlated with growth and the role of the endocardial endothelium as a barrier which controls metabolic exchanges, including the transport of atrial natriuretic factor, between the myocytes and the blood. This endothelial function appears to be essential in the fetal and neonatal age groups since no blood vessels are detected in these groups.     

Sap flow measurements in grapevines (Vitis vinifera L.) 1. Stem morphology and use of the heat balance method

The heat balance method was evaluated in detail for its use in older, mature grapevines with stems of 35 – 45 mm in diameter. Dye colouring of the xylem vessels revealed that even 21 year old grapevines did not show any development of heartwood and that xylem vessels of that age still have the capacity to transport water. A comparison of weight loss of potted vines on a balance and sap flow measurements demonstrated that the heat balance system reflected rapid changes in flow rate without any time delay. However, since even 20 year old xylem vessels of grapevines have the capacity to conduct water, the heater band was not able to heat the sap in all year rings evenly. Apparently, at low flow rates this effect was small and sap flow was calculated correctly. With increasing flow rates large thermal heterogeneities developed upsetting the calculation of the heat balance and mass flow. Consequently, actual sap flow was overestimated by 50 to 100% at high flow rates. This could be attributed to thermal gradients in these relatively thick stems excluding the use of this technique for measurements of long term as well as short term water use patterns in older grapevines. This revised version was published online in June 2006 with corrections to the Cover Date.     

Determination in the system of the neuromuscular relations of the coronary arteries at stages in human postnatal development

By means of micromorphometric method in transversal sections of electively stained right and left coronary arteries the data, characterizing the area of the middle and external sheaths, amount of myelin fibers in nerves, accompanying the artery have been obtained in men of 8 age groups (from birth up to old age), who died from causes not connected with the cardiovascular pathology. The summary amount of all classes of myelin fibers increases synchronously with an increase of the muscle sheath area from newborn up to 35-45 years of age. After this age the amount of the myelin fibers decreases together with decreasing area of the middle sheath. Minimal values of these indices are observed in the old age (85-90 years). The external sheath area increases permanently from birth up to the old age. The correlative analysis reveals a positive and significant connection of various degree between the myelin fibers number and value of the middle arterial sheath in persons of juvenile, mature and elderly age: amount of the muscular layer is adequate to the myelin fibers amount. In other age groups the correlative connections are either small, or not significant. Alternative, corresponding to each age quantitative gradations of the nervous connections are determined by means of the peripheral substrate. Its role in the vessels investigated is performed, to a definite degree, by the muscle sheath.     

Expression of lipoprotein lipase in ovaries of the guinea pig

Guinea pig ovaries were found to have significant lipoprotein lipase (LPL) activity, corresponding to almost one-tenth the activity in paraovarian adipose tissue and in heart per gram of tissue. Northern blot analysis demonstrated the same three species of LPL mRNA in ovaries (1.8, 3.1, and 3.5 kb) as in adipose tissue. In situ hybridization showed LPL mRNA in cells of the follicular wall, and in granulosa and theca lutein cells of the mature corpus luteum. By immunolocalization, LPL was visualized in the vascular endothelium throughout the ovary, but with highest concentration in the endothelium of capillaries and large vessels of the cortical region and capillaries in the stroma of the corpus luteum. These results suggest that in the guinea pig LPL may have a function for the delivery of lipids from lipoproteins to ovarian cells.     

Features of the angioarchitectonics of the papillary muscles and the chordae tendineae of the human heart

In persons of mature, elderly and old age vascularization of the papillary muscles and tendinous chordae in the right and left cardiac ventricles has been studied depending on peculiarities of their structure and sources of their blood supply using injection, macro-microscopical, roentgenoangiographic methods, and silver nitrate impregnation. Certain data concerning the distribution zones in the papillary muscles of the coronary artery branches at various types of the cardiac blood supply are presented. Angioarchitectonics of the papillary muscles are described along their whole extention--in the basal area, in the middle part and at the apex. Architectonic peculiarities of the blood vessels in the muscular-tendinous part of the papillary muscles and in some tendinous chordae are described.     

Vascularization and development of the cytoarchitecture in the human neocortical rudiment

The development of cytoarchitectonics of the brain rudiments in mammals is accompanied by the formation of an intracerebral vascular network. The relationship between these two processes is insufficiently clear. We studied the development of blood vessels and cytoarchitectonics in the neocortical rudiment of 6- to 13-week old human embryos. The light and electron microscopy methods were used, as well as histochemical visualization of NADPH-diaphorase in the vessel cells. The endothelium proliferation was evaluated using antibodies to proliferating cell nuclear antigen. Starting from week 8 of development, the tangentially oriented vessels formed a intraneural network in the ventricular zone of the rudiment, which appears to restrict the motility of neuroepithelial cells. The basal membrane was initially absent, and the neuroepithelial cells were in direct contact with the endothelial cells. During week 9 of development, the tangentially oriented vessels appeared in the intermediate zone. Formations similar to glial legs with short regions of the basal membrane adjoined the walls of inter- and intraneural vessels (note that, according to the published data, glial fibrillary acidic protein is not yet visualized at this stage). Angioarchitectonics depended little on the cell population density in different zones of the rudiment; specifically, the cortical plate did not contain tangentially oriented vessels until week 12-13 of development. The data we obtained suggest that the blood vessels fulfill a special morphogenetic function in the developing neocortex.     

Proliferative activity of blood microvascular endothelium of intestinal mesentery during rat postnatal ontogenesis

The study of mitotic activity of mesenteric microvascular endothelium cells (EC) in 4-, 12-, and 30-day old rats has been carried out using following parameters: number of labeled nuclei per vessel, or per 100 microm of vascular length, or per 1 mm2 of endothelial surface area, as well as shares of labeled EC and of vessels with labeled EC, have been estimated. The highest density of labeled nuclei was revealed in the pericapillary vessels in all rats. Its values were significantly higher in 12-day-old rats and were the lowest in 30-day-old ones.     

The role of the endothelium in the development of functional hyperemia of the skeletal muscles]

Experiments on anesthesized dogs demonstrated that gastrocnemius muscle vessels working hyperemia substantially decreased after chemical destruction of endothelium by saponin, inhibition of endothelium-derived relaxing factor synthesis by gossypol and inhibition of quanylate cyclase by methylene blue. Reaction was not decreased after cyclooxygenase inhibition by indomethacin. The endothelium-derived relaxing factor predecessor--L-arginine essentially increased working hyperemia. We concluded that endothelium plays an important role in reaction of working hyperemia by endothelium-derived relaxing factor release.     

Ultrastructure of the inner surface of the aorta of mature and old animals

In the experiments, performed on 12 white rats and 8 rabbits, by means of scanning electron microscopy of the native preparations and in a number of cases with use of silver nitrate impregnation, the internal surface structure has been studied in the aortal membrane of mature and old animals. At ageing the integrity and continuity of the endothelial monolayer is preserved, on the surface local intimal pits, craters and microdefects appear, adhesiveness of endothelium to leucocytes increases. Orientation of the intimal folds is disturbed. The type of the senescent remodeling in the endothelial layer revealed predisposes to development of atherosclerosis.