Densitatea microvaselor în peretele venei safene mari la pacienții cu afecțiuni metabolice asociate

Background. The media of vessels is virtually avascular, oxygen and nutrients reach the media by diffusion. The vessels of the media appear mainly from the adventitia. Objective of the study. To investigate the density of vasa vasorum in the wall of great saphenous vein and to evaluate the influence of the atherosclerosis and diabetes mellitus on it. Material and Methods. Great saphenous vein specimens (n=9) were collected from the patients whose lower limbs were amputated: crush-syndrome (CS), atherosclerosis (ATS), atherosclerosis associated with diabetes mellitus (ATS+DM). The speciments were double immunostaining for CD34 and α-SMA. Results. Immunohistochemical quantification revealed deeper penetration of vasa vasorum in the tunica media in all evaluated cases. In case of the CS the vessels of the adventitia (165 vessels/mm2) prevailed numerically those of the media (140 vessels/mm2). In case of the ATS the values of the vascular density were marked both in the adventitia (246±101 vessels/mm2) and in the media (250±130 vessels/mm2). In case of the ATS+DM the vessel density of the media (323±104 vessels/mm2) exceeded numerically the density of the adventitia (173±29 vessels/mm2). Conclusion. The increased microvessel density of the great saphenous vein may be induced by stimuli other than vessel wall thickness, including atherosclerosis and diabetes mellitus. Introducere. Media vaselor este practic avasculară, oxigenul și nutrienții ajungând la ea prin difuzie. Vasele mediei apar prin proliferare preponderent din vasele adventicei. Scopul lucrării. De a cerceta densitatea vasa vasorum în peretele venei safene mari și de a evalua influența aterosclerozei și a diabetului zaharat asupra ei. Material și Metode. Au fost recoltate fragmente de venă safenă mare (n=9) de la pacienții ale căror membre inferioare au fost amputate: crush-syndrome (CS), ateroscleroză (ATS), ateroscleroză și diabet zaharat asociat (ATS+DZ). Fragmentele venoase au fost dublu colorate cu antiCD34 și anti-(α)SMA. Rezultate. Cuantificarea imunohistochimică a relevat o penetrare profundă a vasa vasorum în tunica medie în toate cazurile evaluate. În cazul pacientului cu CS numeric au prevalat vasele adventicei (165 vase/mm2) în raport cu cele ale mediei (140 vase/mm2). În cazul pacienților cu ATS, valorile densității vasculare erau marcante atât la nivel de adventice (246±101 vase/mm2), cât și la nivel de medie (250±130 vase/mm2). În cazurile de ATS+DZ, densitatea vaselor mediei (323±104 vase/mm2) depășea numeric densitatea celor din adventice (173±29 vase/mm2). Concluzii. Creșterea densității microvaselor venei safene mari poate fi indusă de alți stimuli decât grosimea vasului, printre care pot fi ateroscleroza și diabetul zaharat

Histopathological and immunohistochemical peculiarities of varicose veins of the lower limbs

Introducere. Boala varicoasă sau boala venoasă cronică poate afecta toate segmentele sistemului venos, dar interesează cu precădere sistemul venei cave inferioare. Mai frecvent, leziunile afectează venele superficiale ale membrelor inferioare. Deşi, au fost publicate numeroase lucrări asupra modificărilor morfologice ale venelor afectate de boala varicoasă, este încă neclar care este succesiunea acestor leziuni şi cum contribuie la apariţia varicelor. Scopul acestui studiu a fost de a clarifica schimbările histopatologice şi imunohistochimice prezente de-a lungul unei vene safene dilatate varicos şi în venele safene la diferite etape evolutive ale bolii venoase. Material şi metode. Drept obiect de sudiu au servit ve¬nele safene dilatate varicos, înlăturate chirurgical în cadrul safenectomiilor. Materialul postoperator a fost colectat de la 8 pacienţi cu vârsta cuprinsă între 19 şi 68 de ani, internaţi pentru tratament chirurgical în secţia de chirurgie vasculară a SCR „Timofei Moşneaga” din Chişinău. Conform clasificării CEAP a bolii venoase cronice, lotul de pacienţi a fost divizat în două grupuri: sublotul 1-5 pacienţi în stadiile clinie CEAP2_3, sublotul 2-3 pacienţi în stadiile clinice CEAP4_6. Venele varicoase au fost fixate în formalină neutră, incluse în parafină şi colorate prin metode de colorare uzuale, specifice şi imunohistochimice. Rezultate. Modificările histopatologice ale peretelui venos (sublot 1) au cuprins îngroşarea mediei şi intimei din contul hipertrofiei şi hiperplaziei celulelor musculare netede, atât la nivel subendotelial, cât şi în tunica medie. Creşterea compo-nentei fibrilare colagenice a fost asociată cu creşterea componentei fibrilare elastice. Venele sublotului 2 aveau intima şi media îngroşate datorită prezenţei colagenului în cantităţi mari. Fasciculele de miocite ale mediei au fost fragmentate şi înlocuite de fibrele de colagen în exces. Sublotul 2 a prezentat, de ase-menea, detaşarea endoteliului şi înlocuirea stratului endotelial cu fibre de colagen. Adventicea, la rândul său, a prezentat un număr impunător de vase ale microcirculaţiei (yasa vasorum). Concluzii. În stadiile clinice incipiente ale bolii venoase cronice (sublot 1) predomină hiperplazia şi hipertrofia celulelor musculare netede ale mediei. În stadiile clinice avansate (sublot 2) se declanşează un proces de fibrozare a mediei şi intimei (fleboscleroză), denudarea endoteliului conduce, de obicei, la formarea trombilor în lumenul venei (tromboflebită), iar densitatea microcirculaţiei la nivelul adventicei creşte odată cu avansarea bolii.Introduction. Varicose disease or chronic venous disease may affect all segments of the venous system but particularly affects the system of the inferior vena cava. More frequently, the lesions affect superficial veins of the lower limb. Although there were published numerous works about morphological modifications of veins affected by the varicose disease it is still development of varicose veins. The purpose of the current research was to clarify histopathological and immunohistochemical modifications present along a single saphenous vein or in the saphenous veins at various evolutionary stages of the venous disease. Material and methods. As object of studying served the varicose saphenous veins surgically removed during saphenoustomy. Postoperative material was collected from eight patients aged from 19 to 68 years and admitted for surgical treatment at the Department of vascular surgery of "Timofei Moşneaga” Republican Clinical Hospital, Chisinau, Republic of Moldova. According to CEAP classification of the chronic venous disease, the patients were divided in two subgroups: subgroup 1 - five patients at clinical stages CEAP2-3 and subgroup 2 - three patients at clinical stages CEAP4-6 The varicose veins were fixed in formalin, embedded in paraffin and stained using routine, special and immunohistochemical staining methods. Results. Histopathological modifications of venous wall (subgroup 1) comprise thickening of media and intima due to the hypertrophy and hyperplasia of smooth muscle cells both of subendothelium layer and media. Increasing fibrillar collagen components was associated with increasing fibrillar elastic components. Veins of subgroup 2 had thick media and intima because of presence of large amount of collagen. Sub¬group 2 showed detachment of endothelium and it replacement with fibres of collagen. Adventitia, in turn, presented a large number of microcirculatory vessels (vasa vasorum). Coclusions. At the incipient clinical stages of the chronic venous disease predominate the hypertrophy and hyperplasia of smooth muscle cells of the media. At the advanced stages a process of sclerosis of media and intima starts (phlebosclerosis). Endothelium detachment leads to the thrombus formation in the venous lumen (thrombophlebitis), but the intensity of microcirculation at the level of adventitia increases together with the evolution of the disease

Immunohistochemical Localization of Dendritic Cells in Human Dental Pulp

Svrha: Svrha istraživanja bila je metodom imunohistokemijskih bojenja locirati dendritičke stanice, te dio imunokompetentnog sustava dentalne pulpe u histološkim uzorcima normalne i ireverzibilno inflamirane ljudske zubne pulpe. Materijal i metode: Zbog posebnih svojstava vezivnog tkiva ljudske zubne pulpe, koristila su se sljedeća antitijela: vimentin V9, protein povezan s neurofi lamentima (NFAP), protein S 100 te aktin glatkih mišića (klon 1A4). Rezultati: Rezultati su pokazali da humana dentalna pulpa ima mnogobrojne stanice s razgranatim citoplazmatskim produžecima u području upalnoga infiltrata. Na temelju imunohistokemijskog bojenja smatramo da je riječ o dendritičkim stanicama. Zaključak: Dendritičke stanice u upaljenoj ljudskoj zubnoj pulpi nisu neočekivane s obzirom na veliku gustoću upalnog infiltrata i ulogu koju imaju u upalnom odgovoru. Podtip i značenje tih stanica treba još objasniti.Objectives: The aim of this study was to locate the dendritic cells, a part of the immunocompetent system of dental pulp, in histological samples of normal and irreversibly inflamed human dental pulp, using immunohistochemical staining methods. Material and Methods: In light of the particular character of the connective tissue of human dental pulp, the following antibodies were used: vimentin clone V9, a protein associated with neurofilaments (NFAP), the S 100 protein, and smooth muscle type actin (clone 1A4). Results: The results indicated that human dental pulp has numerous cells with branching cytoplasmic extensions, in areas of inflammatory infiltrate. Based on immunohistochemical staining we consider these to be dendritic cells. Conclusions: The presence of dendritic cells in inflamed human dental pulp is not unexpected, considering the great density of the inflammatory infiltrate, and the role they play in the inflammatory response. The subtype and significance of these cells need to be clarified

Immunohistochemical Localization of Dendritic Cells in Human Dental Pulp

Svrha: Svrha istraživanja bila je metodom imunohistokemijskih bojenja locirati dendritičke stanice, te dio imunokompetentnog sustava dentalne pulpe u histološkim uzorcima normalne i ireverzibilno inflamirane ljudske zubne pulpe. Materijal i metode: Zbog posebnih svojstava vezivnog tkiva ljudske zubne pulpe, koristila su se sljedeća antitijela: vimentin V9, protein povezan s neurofi lamentima (NFAP), protein S 100 te aktin glatkih mišića (klon 1A4). Rezultati: Rezultati su pokazali da humana dentalna pulpa ima mnogobrojne stanice s razgranatim citoplazmatskim produžecima u području upalnoga infiltrata. Na temelju imunohistokemijskog bojenja smatramo da je riječ o dendritičkim stanicama. Zaključak: Dendritičke stanice u upaljenoj ljudskoj zubnoj pulpi nisu neočekivane s obzirom na veliku gustoću upalnog infiltrata i ulogu koju imaju u upalnom odgovoru. Podtip i značenje tih stanica treba još objasniti.Objectives: The aim of this study was to locate the dendritic cells, a part of the immunocompetent system of dental pulp, in histological samples of normal and irreversibly inflamed human dental pulp, using immunohistochemical staining methods. Material and Methods: In light of the particular character of the connective tissue of human dental pulp, the following antibodies were used: vimentin clone V9, a protein associated with neurofilaments (NFAP), the S 100 protein, and smooth muscle type actin (clone 1A4). Results: The results indicated that human dental pulp has numerous cells with branching cytoplasmic extensions, in areas of inflammatory infiltrate. Based on immunohistochemical staining we consider these to be dendritic cells. Conclusions: The presence of dendritic cells in inflamed human dental pulp is not unexpected, considering the great density of the inflammatory infiltrate, and the role they play in the inflammatory response. The subtype and significance of these cells need to be clarified

Paul Langerhans: hodočasnik koji je "putovao" of funkcionalne histologije do biologije mora

The nineteenth century was the time of a real revolution in science and medicine. A lot of seminal discoveries in medicine and biology were done in this time, and many of them were coincident with the introduction of the compound microscope by Hermann van Deijl and the standard histological technique by Paul Ehrlich. The main tissue types and individual cells were characterized and originally classified more than hundred years ago, although less attention was paid to their basic functions. This was mainly due to the modality of tissue specimen processing that allowed particularly detailed descriptive studies. Even so, we can notice some attempts to correlate the structure with the function. The German scientist Paul Langerhans, well-known for the discovery of Langerhans islets of the pancreas and Langerhans cells from the epidermis, tried to change the conventional fate of morphological studies introducing in his works functional hypothesis based on traditional microscopic observations even from the beginning of his scientific career. Paul Langerhans was a complex personality of the second half of the nineteenth century, not only in medicine, but also in other fields of biology. In the present review, presented is the life and research activity of Paul Langerhans, not only because of the importance of his discoveries, but also for perspectives that were opened by these findings in unexpected fields of medicine and biology.Devetnaesto stoljeće bilo je vrijeme prave revolucije u znanosti i medicini. U tom je stoljeću došlo do mnogih otkrića u medicini i biologiji, a mnoga od njih su se podudarala s primjenom mikroskopa (Hermann van Deijl) i uvođenjem standardne histološke tehnike (Paul Ehrlich). Glavne vrste tkiva i pojedinačne stanice karakterizirane su i izvorno klasificirane prije više od stotinu godina, iako je manje pozornosti bilo posvećeno njihovim osnovnim funkcijama. To je uglavnom bilo zbog modaliteta obrade uzoraka tkiva koja je omogućavala posebno detaljne deskriptivne studije. Ipak, primjećuju se neki pokušaji povezivanja strukture s funkcijom. Njemački znanstvenik Paul Langerhans, poznat po otkriću Langerhansovih otočića gušterače i Langerhansovih stanica u epidermisu, pokušao je promijeniti konvencionalnu sudbinu morfoloških istraživanja uvodeći već od početka svoje znanstvene karijere u svoja istraživanja funkcionalnu hipotezu koja se temelji na tradicionalnim mikroskopskim promatranjima. Paul Langerhans bio je složena ličnost druge polovice XIX. stoljeća ne samo u medicini nego i u drugim područjima biologije. U ovom radu predstavljen je njegov život i njegova istraživačka djelatnost ne samo zbog važnosti njegovih otkrića već i zbog gledišta koja su ta otkrića potaknula u neočekivanim područjima medicine i biologije

Immunoexpression of mast cells and macrophages in angiogenesis of atherosclerotic plaques in patients with metabolic syndrome

This article is the result of our long-term studies in the area of atherogenesis and continuing to date search for answers to unresolved issues, in cooperation with the Angiogenesis Research Center, the Victor Babes University of Medicine and Pharmacy (Timisoara, Romania). The results of these studies were presented at many national and international symposia and congresses. Numerous studies have demonstrated that endothelial damage is a precursory symptom of atherosclerosis, which leads to an increase of vascular permeability, activation of mast cells and migration of leukocytes, lymphocytes, macrophages, adhesion of platelets, proliferation of vascular smooth muscle cells and eventual vasospasm and proinflammatory condition. All of the above listed components can be rightfully considered active pathogenetic participants in atherosclerosis and a result of aggregation of all risk factors that accompany a wide variety of cardiovascular diseases, such as coronary heart disease, hypertension, diabetes, dyslipidemia, etc. The influx of monocytes and mast cells during the early stages of atherosclerosis leads to the most pronounced manifestations of vascular inflammation, especially in patients with metabolic disorders [1]. Angiogenesis is a very important pathogenetic element of atherosclerosis in stages of complicated plaques, along with mast cells and macrophages. CD-105 is a sensitive marker of newly formed endothelial cells, an effective index of activation and proliferation of microvessels, not only in aggressive forms of cancer, but also in atherosclerotic plaques of the affected vessels. The plaque neovascularization process often begins in intima, progresses and leads to further destabilization of atherosclerotic plaques (intramural hemorrhage, ruptures etc.). Also, anti-MCT (mast cell tryptase) and CD-68 demonstrate clearly the important pathogenetic stages and patterns of atherosclerosis development and its complications in patients with metabolic disorders. In our study, we analyzed the histotopographic distribution of newly formed blood vessels as a feature of angiogenesis, the extent of mast cell degranulation, the expression of macrophages in different types of plaques, as well as various arterial vessels in patients with atherosclerosis and metabolic syndrome, complicated by atherosclerosis. We have tried to analyze the importance of mast cells and macrophages, the patterns of development of atherosclerosis stages, along with diagnostic and prognostic features. The study included 34 patients, who died of atherosclerosis (no. =17) and atherosclerotic complications of metabolic syndrome (no. =17). Fragments of their cerebral (middle cerebral arteries), carotid, coronary arteries, aorta (thoracic and abdominal segments), renal, iliac and vertebral arteries were collected for research at autopsy. The fragments were processed using standard techniques. The type definition of plaques was based on morphological classification, as well as on macroscopic and histological images of hematoxylin-eosin stained sections and on histochemical methods – silver and orcein impregnation. To determine the expression of mast cells in the affected vessels, we have used anti-MCT immunohistochemical stain. Macrophages were identifi ed using the CD-68 specifi c marker and the newly formed vessels – respectively, by using CD- 105 (Endoglin), which is specific. The evaluation of the results was based on determining the density and intensity of the final reaction, reflected in the quantitative ratio of different zones of atheromatous plaques. Positively stained mast cells, macrophages and newly formed vessels were found in many types of atherosclerotic plaques, especially in adventitia and in the immediate vicinity of plaques and in subendothelial layers. We found a statistical correlation between the plaque type and clinical data. The immunohistochemical method is effective for determining mast cells, macrophages, and newly formed vessels of atherosclerotic plaques, directly refl ecting many important pathogenetic elements of atherogenesis in patients with metabolic syndrome

Immunohistochemical expression of androgen receptor and prostate-specific antigen in breast cancer.

AR (androgen receptor) and PSA (prostate-specific antigen) are involved in the pathogenesis of breast cancer, but their role is not clearly defined. The purpose of this study was to analyze by immunohistochemistry the AR and PSA (prostate-specific antigen) expression in 156 female breast carcinomas and to correlate the results with some histopathological parameters, like ER (estrogen receptor), PR (progesterone receptor), HER2/neu, nodal and metastasis status, histological type and grade. ARs and PSA were expressed in 112/156 (72%) and respectively in 61/156 (39%) of cases and we found a positive correlation between AR and PSA expression in breast carcinomas (