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Biomedical subjects

E Mandache

Publications and source records attributed to E Mandache.

At least 37 records · Page 2Linked to original sources

Accessory cells and macrophages in the histopathology of SIVsm-infected cynomolgus monkeys.

Thirty-three out of 39 cynomolgus monkeys (Macaca fascicularis) infected with SIVsm (strain SMM-3) developed various pathologies similar to those seen in human AIDS. Lymphadenopathy was frequently seen (72%) and was characterized by hyperplasia followed by involution of follicle/germinal centres due to follicular dendritic cell (FDC) destruction corresponding to the degree of immunodeficiency. Various organs such as the lungs, liver, central nervous system, kidneys, gastrointestinal tract, cardiovascular system and adrenals showed histopathological changes with prominent monocyte/macrophage and multinucleated giant cell formation. Eighteen (54%) monkeys presented with extranodal malignant lymphoma (ML) associated with marked CD4 decrease and destruction of follicular architecture. The high frequency of ML, giant cell disease and lymph node changes seen in the present SIV model provides an attractive system to elucidate the role of FDC and monocytes/macrophages in the pathogenesis of these conditions in common with HIV infection and human AIDS.

Animals↗

Ultrastructural defects of the glomerular basement membranes associated with primary glomerular nephropathies.

It is generally accepted that a glomerular basement membrane (GBM) thinner than 200 nm should be considered below normal. When this abnormality has a global and diffuse distribution, the associated clinical condition is a benign familial hematuria related to mutations of the COL4A4/COL4A3 genes, or an Alport syndrome. More often the GBM defects display a focal and segmental pattern, too small to express a thin glomerular basement membrane disease. The aim of this study is to emphasize statistical data concerning the pathogenic link between the renal glomerular diseases and the preexisting thin and very thin GBM. A series of 487 renal biopsies from adult patients has been thoroughly investigated both for nephropathologic diagnosis and the GBM ultrastructure. It has been statistically concluded that there is a close coexistence of primary glomerulonephritis and thin glomerular basement membranes with the role of a predisposing condition for immune complex deposition.

Adolescent↗

Neoductular progenitor cells regenerate hepatocytes in severely damaged liver: a comparative ultrastructural study.

In severely injured liver, stem cells give rise to progeny that tend to replace lost hepatocytes. Neoductular reaction appears as an inherent stage of liver reconstruction following severe damage caused by different pathological mechanisms. Few ultrastructural types of progenitor cells have been described, and some molecular phenotypes of progenitor stages have been characterized, but the details of the differentiation process are largely unknown. We prepared for light and electron microscopy examination human liver from biopsies of patients with chronic active hepatitis, and rat liver with allyl alcohol-induced periportal necrosis. We found that progenitor neoductular cells acquire the hepatocytic polarity pattern during a multi-step process apparently involving cell migration and dissolution of neoductular basement membrane. An intermediate stage with "mixed" ductular and hepatocytic polarity was described.

Animals↗

The predictive value of peritubular capillaries C3d deposition in IgA glomerulonephritis.

The IgA glomerulonephritis (IgAGN) is one of the most common primary glomerulonephritis and has a variable and difficult to predict evolution toward the end-stage nephrosclerosis. The deposition of C3d complement component in peritubular capillaries (PTCs) indicates a variant type of acute rejection while C3d deposition in primary glomerulonephritis (GN) is poorly documented. The aim of this study is to examine C3d expression in peritubular capillaries (PTCs) as a predictive marker and its correlation with the severity of renal injury in IgA glomerulonephritis. Polyclonal FITC conjugated rabbit anti-human C3c and C3d antibodies were used for direct immunofluorescent evaluation of the C3c and C3d deposits in 24 kidney biopsies with IgA glomerulonephritis. The study revealed that the C3d deposits in peritubular capillaries were associated with known predictors for rapid progression of IgAGN: glomerular sclerosis (63.6%), atrophic tubules (90.9%) and interstitial sclerosis (81.8%). The intensity of the C3c glomerular immunofluorescent deposits was related with active lesions. Thus, the predictive value of C3d deposition on PTCs in IgAGN is worth to be taken into consideration as an unfavorable outcome of the disease and request further long run investigations.

Adolescent↗

Interstitial Cajal-like cells (ICLC) in atrial myocardium: ultrastructural and immunohistochemical characterization.

We have previously reported (Hinescu & Popescu, 2005) the existence of interstitial Cajal-like cells (ICLC), by transmission electron microscopy, in human atrial myocardium. In the present study, ICLC were identified with non-conventional light microscopy (NCLM) on semi-thin sections stained with toluidine blue and immunohistochemistry (IHC) for CD117/c-kit, CD34, vimentin and other additional antigens for differential diagnosis. Quantitatively, on semi-thin sections, ICLC represent about 1-1.5% of the atrial myocardial volume (vs. approximately 45% working myocytes, approximately 2% endothelial cells, 3-4% for other interstitial cells, and the remaining percentage: extracellular matrix). Roughly, there is one ICLC for 8-10 working atrial myocytes in the intercellular space, beneath the epicardium, with a characteristic (pyriform, spindle or triangular) shape. These ICLC usually have 2-3 definitory processes, emerging from cell body, which usually embrace atrial myocytes (260 nm average distance plasmalemma/sarcolemma) or establish close contact with nerve fibers or capillaries (approximately 420 nm average distance to endothelial cells). Cell prolongations are characteristic: very thin (mean thickness = 0.15+/-0.1 microm), very long for a non-nervous cell (several tens of microm) and moniliform (uneven caliber). Stromal synapses between ICLC and other interstitial cells (macrophages) were found (e.g. in a multicontact type synapse, the average synaptic cleft was approximately 65 nm). Naturally, the usual cell organelles (mitochondria, smooth and rough endoplasmic reticulum, intermediate filaments) are relatively well developed. Caveolae were also visible on cell prolongations. No thick filaments were detected. IHC showed that ICLC were slightly and inconsistently positive for CD117/c-kit, variously co-expressed CD34 and EGF receptor, but appeared strongly positive for vimentin, along their prolongations. Some ICLC seemed positive for a-smooth muscle actin and tau protein, but were negative for nestin, desmin, CD13 and S-100. In conclusion, we provide further evidence of the existence of ICLC in human atrial myocardium, supporting the possible ICLC role in pacemaking, secretion (juxta- and/or paracrine), intercellular signaling (neurons and myocytes). For pathology, ICLC might as well be 'players' in arrhythmogenesis and atrial remodeling.

Animals↗

Insights into the interstitium of ventricular myocardium: interstitial Cajal-like cells (ICLC).

We have previously described interstitial Cajal-like cells (ICLC) in human atrial myocardium. Several complementary approaches were used to verify the existence of ICLC in the interstitium of rat or human ventricular myocardium: primary cell cultures, vital stainings (e.g.: methylene blue), traditional stainings (including silver impregnation), phase contrast and non-conventional light microscopy (Epon-embedded semithin sections), transmission electron microscopy (TEM) (serial ultrathin sections), stereology, immunohistochemistry (IHC) and immunofluorescence (IF) with molecular probes. Cardiomyocytes occupy about 75% of rat ventricular myocardium volume. ICLC represent approximately 32% of the number of interstitial cells and the ratio cardiomyocytes/ICLC is about 70/1. In the interstitium, ICLC establish close contacts with nerve fibers, myocytes, blood capillaries and with immunoreactive cells (stromal synapses). ICLC show characteristic cytoplasmic processes, frequently two or three, which are very long (tens up to hundreds of microm), very thin (0.1-0.5 microm thick), with uneven caliber, having dilations, resulting in a moniliform aspect. Gap junctions between such processes can be found. Usually, the dilations are occupied by mitochondria (as revealed by Janus green B and MitoTracker Green FM) and elements of endoplasmic reticulum. Characteristically, some prolongations are flat, with a veil-like appearance, forming a labyrinthic system. ICLC display caveolae (about 1 caveola/ 1 microm cell membrane length, or 2-4% of the relative cytoplasmic volume). Mitochondria and endoplasmic reticulum (rough and smooth) occupy 5-10% and 1-2% of cytoplasmic volume, respectively. IHC revealed positive staining for CD34, EGFR and vimentin and, only in a few cases for CD117. IHC was negative for: desmin, CD57, tau, chymase, tryptase and CD13. IF showed that ventricular ICLC expressed connexin 43. We may speculate that possible ICLC roles might be: intercellular signaling (neurons, myocytes, capillaries etc.) and/or chemomechanical sensors. For pathology, it seems attractive to think that ICLC might participate in the process of cardiac repair/remodeling, arrhythmogenesis and, eventually, sudden death.

Animals↗

Caveolae in smooth muscles: nanocontacts.

Smooth muscle cell (SMC) caveolae have been investigated by quantitative and qualitative analysis of transmission electron microscopy (TEM) images of rat stomach, bladder and myometrium, guinea pig taenia coli, human ileum, and rat aortic SMCs. Ultrathin (below 30 nm) serial sections were used for examination of caveolar morphology and their connections with SMC organelles. Average caveolar diameter was smaller in vascular SMCs (70 nm, n=50) than in visceral SMCs (77 nm, n=100), but with the same morphology. Most of the caveolae, featured as flask-shaped plasma membrane (PM) invaginations, opened to the extracellular space through a 20 nm stoma (21 +/- 3 nm) having a 7 nm thick diaphragm. A small percentage of caveolae (3%), gathered as grape-like clusters, did not open directly to the extracellular space, but to irregular PM pockets having a 20-30 nm opening to the extracellular space. In visceral SMCs, caveolae were disposed in 4-6 rows, parallel to myofilaments, whilst aortic SMCs caveolae were arranged as clusters. This caveolar organization in rows or clusters minimizes the occupied volume, providing more space for the contractile compartment. The morphometric analysis of relative volumes (% of cell volume) showed that caveolae were more conspicuous in visceral than in vascular SMCs (myometrium - 2.40%; bladder - 3.66%, stomach - 2.61%, aorta - 1.43%). We also observed a higher number of caveolae per length unit of cellular membrane in most visceral SMCs compared to vascular SMCs (myometrium - 1.06/microm, bladder - 0.74/microm, aorta - 0.57/microm, stomach - 0.48/microm). Caveolae increase the cellular perimeter up to 15% and enlarge the surface area of the plasma membrane about 80% in SMCs. Threedimensional reconstructions (15micro(3)) showed that most caveolae, in both visceral and vascular SMCs, have nanocontacts with SR (87%), other with mitochondria (10%) and 3% apparently have no contact with these organelles. Usually, 15 nm wide junctional spaces exist between caveolae and SR, some of them with nanostructural links between each other or with mitochondria: direct contacts (space <2 nm or none) and molecular links, so called 'feet' (about 12 nm electron dense structures between organellar membranes). Direct contacts possibly allow molecular translocation between the two membranes. Electron-dense 'feet'-like structures suggest a molecular link between these organelles responsible for intracellular Ca(2+) homeostasis (excitation-contraction coupling or pharmaco-mechanical coupling). Close appositions (approximately 15 nm) have also been observed between caveolae and perinuclear SR cisternae, suggesting that caveolae might be directly implicated in excitation-transcription coupling.

Animals↗

The Kupffer cells. Ultrastructural and functional features.

The Kupffer cells, as fixed macrophages, belong to the mononuclear phagocyte system (MPS), the former reticulo-endothelial system (RES). Their ultrastructure has some characteristics, i.e. ergastroplasmic peroxidase activity and worm-like structures, which allow a very sharp identification. Some particular "bridge-like" arrangements of Kupffer cells in the capillary lumen seem to provide them with a special ability of "trapping" the passing white blood cells. The important endocytotic function is analysed from the viewpoint of ultrastructural features, and with particular reference to the phagocytosis of damaged red blood cells. Several drugs and particulate substances acting as Kupffer cells stimulants are presented. This stimulation reaction concerns woth increased number and enhanced endocytotic activity. The origin and kinetics of Kupffer cells are discussed, with reference to own results which substantiate the theory of local proliferation and blood supply, the bone marrow being however the main source of hepatic macrophages. A few ultrastructural data on Kupffer cells involved in pathological processes are presented.

Acid Phosphatase↗

Ultrastructural features in an erythrophagocytic T4 lymphoma.

Electron microscopic investigation allowed the identification and characterization of the atypical malignant cells in a case of T-cell lymphoma. The proliferating cells were endowed with characteristics belonging to two different lineages: phagocytic ability, complement receptors and ultrastructural features proper to the macrophagic lineage, and T-cell determinants (E receptors, T3, T4 and T11 antigens). The cells were peroxidase and esterase negative. The erythrocytes were partially or completely dehemoglobinized and presented the phenomenon of autolysis in different stages of evolution. Because this lymphoma is difficult to diagnose and is apparently resistant to therapy, its recognition and further study are warranted.

Biomarkers, Tumor↗

The ultrastructure of the abdominal lymph nodes cortex in rats during primary and secondary immune response (a special reference to dendritic reticulum cells and interdigitating cells).

The stimulation of coeliac rat lymph nodes was performed by intraperitoneal injections of typhoid vaccine and was unique for the primary immune response and repeated after 6 weeks for the secondary response. The light and electron microscopic observations showed for the primary response, an early germinal center reaction, which might be accounted for by a background of continuous stimulation of the coeliac nodes, stemming from the digestive tract. The dendritic reticulum cells (DRC), considered typical for the B area, were located at the borderline between the germinal center and the mantle zone. Their cytoplasmic extensions penetrated the lymphocyte-lymphoblastic center, surrounding most of the germinal center cells. The marginal zone and the paracortex reacted as a whole, the interdigitating cells (IDC) being the dominant feature. An explanation would be that the marginal zone can be penetrated by T cells and connected IDCs, thus, the B and T areas seem to be largely interspersed. The results suggest that IDCs are cells of direct monocytic origin.

Abdomen↗

The involvement of omentum and its milky spots in the dynamics of peritoneal macrophages.

The investigation has been carried out on stimulated and unstimulated peritoneal cavities of rats. China ink and Corynebacterium parvum were injected i.p. both as peritoneal stimuli and markers. Omenta were picked up at time intervals beginning with 10 min and up to seven days after the i.p. injection. The light and electronmicroscopic investigation showed after 10-30 minutes labeled macrophages stuck as monolayers on some peritoneal areas corresponding to the milky spots which developed in size and number. Days after the i.p. injection the labeled macrophages were found deeper in the milky spots. After the fourth day they appeared in the regional lymph nodes. The milky spots contained also large lymphocytes and plasma cells. The results suggest that milky spots are not only places of resident macrophages development and release in the peritoneal cavity but also their exit pathways. Therefore the omentum leads the traffic of peritoneal macrophages. The developed milky spots play also the role of lymphoid structures providing grounds for macrophage-lymphocyte contacts.

Animals↗

The development of lymphatic follicles in the omentum after intraperitoneal stimulation of rats.

China ink and typhoid vaccine were used as peritoneal stimuli injected in single doses. The subsequent omental reaction was studied after several time intervals: 6 and 12 hours, 1, 2, 3, 4 and 8 days, 3 and 6 weeks. The omental reaction featured as lymphatic nodules connected to milky spots was compared with a primary immune response. The ultrastructure and development of the characteristic cells (lymphocytes, lymphoblasts, dendritic reticulum cells, tingible body macrophages and plasma cells) was investigated throughout the period from 6 h to 6 weeks. A special attention was devoted to ultrastructural and functional features of plasma cells.

Animals↗