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P Preda

Publications and source records attributed to P Preda.

35 records · Page 2Linked to original sources

Healing of prosthetic arterial grafts.

Numerous synthetic biomaterials have been developed as vascular substitutes. In vitro, ex vivo and in vivo studies have demonstrated that in animals, selected materials, i.e., Dacron and ePTFE (expanded polytetrafluoroethylene) grafts, are successfully incorporated in both the large and the small caliber host arteries through a process which is generally referred to as graft healing. Morphologically, this process consists of a series of complex events including fibrin deposition and degradation, monocyte-macrophage recruitment and flow-oriented cell-layer generation, this last event being the complete endothelialization of the arterial substitute. In contrast to experimental animals, the flow surface of synthetic vascular grafts remains unhealed in humans, particularly in the small caliber conduits. Healing in man consists of graft incorporation by the perigraft fibrous tissue response with a surface covered by more or less compacted, cross-linked fibrin. It is therefore obvious that: i) marked differences in graft healing exist between animals and man; and ii) the usual mechanisms of graft endothelialization are partially ineffective in man. In order to guarantee the patency of synthetic vascular grafts for human small artery bypass, new strategies and approaches have recently been attempted. In particular, the endothelial cell seeding approach has been successfully accomplished in animals and is being experimented in human clinical studies. The problems and results of this biological approach are outlined in this paper.

Animals↗

An ultrastructural and immunocytochemical analysis of human endothelial cell adhesion on coated vascular grafts.

Human adult endothelial cells were enzymatically harvested from adipose tissue. Cell viability was established by Trypan blue exclusion and transmission and scanning electron microscopy. Endothelial cells were identified by immunocytochemical investigation at light microscopy, transmission electron microscopy, and scanning electron microscopy. Isolated cells were positive for actin and vimentin, negative for desmin. Factor VIII RA was mainly expressed at cell surface and occasionally disclosed in the cytoplasm. Reactivity for UEA I and J15 was weak or undetectable. Human endothelial cells were seeded and left to adhere for one hour onto different nonvascular substrates (glass, poly-l-lysine, formvar-carbon, fibronectin, Teflon). Scanning electron microscopy defined surface features, suggesting tenacious cell adhesion on the substrate. Different vascular substrates were tested (preclotted Dacron, albumin Dacron, Hemashield Dacron, Gelseal Dacron, ePTFE, fibronectin-ePTFE). Commercially available coated grafts showed qualitative and quantitative differences in cell adhesion. In particular, Gelseal Dacron provided the best quantitative results, even though a wide variability was observed. In contrast, fibronectin-coated ePTFE gave more reliable results and high spreading efficiency. In the short term, coated grafts do not seem to offer greater advantages than fibronectin-coated ePTFE. However, specific incubation times for each coated graft should be selected and the long-term approach (graft culture) should also be attempted.

Adipose Tissue↗

Electron microscopy of lipid deposits in human atherosclerosis.

The filipin probe associated with tannic acid stain was used to study intra- and extracellular lipids in surgically removed human atherosclerotic lesions (n = 20). In particular, intimal thickenings, fatty streaks and fibrolipidic plaques have been investigated by using mainly transmission and scanning electron microscopy. In the intimal thickenings, the lipid deposits were mainly localized in the subendothelial space as homogeneously sized particles (40-140 nm) and more heterogeneous uni-multilamellar vesicles (35-700 nm). Intermediate lipid forms were also observed. In the fatty streaks, the lipid deposits were intracellular and mainly observed in cells with a monocyte/macrophagic phenotype. Lipid inclusions, lipid lysosomal bodies and intracellular cholesterol crystals very similar to those observed in experimentally induced atherosclerosis were documented. In the fibrolipidic plaque the lipid deposits were found both in the intracellular and in the extracellular compartments. Lipids accumulated within arterial macrophages and smooth muscle cells, usually as lipid droplets. Clusters of lipoprotein-like particles (50 nm in diameter) as well as larger uni-multilamellar lipids (700 nm) with an occasional compound appearance were particularly observed bound to elastic tissue and collagen fibers. These morphological observations outline the complexity of lipid metabolism in the various histological aspects of human atherosclerosis.

Aged↗

Endothelial injury in human atherosclerosis.

A light and electron microscopic investigation (scanning and transmission electron microscopy) was performed on 51 human atherosclerotic carotid lesions. The purpose of this study was to establish whether features of endothelial injury such as those described in animals occur in man and whether these features can be related to specific stages of human atherosclerosis. Irrespective of their histological appearance the atherosclerotic lesions were covered with endothelium which showed non-specific changes in cell shape and size. However, all complicated lesions appeared denuded. Moreover, a peculiar interaction of endothelium with monocytes and lymphocytes as well as blood components (e.g., fibrin and lipoproteins) was observed in intimal thickenings, fatty streaks and uncomplicated plaques. The surface exposure of macrophage-derived foam cells was seen on florid fatty lesions. Large areas of the arterial surface lacking any endothelial coverage were characteristic of complicated plaques. They appeared to be a consequence of the arterial wall degeneration with an associated failure in endothelial repair.

Arteriosclerosis↗

Endothelialization of a new Dacron graft in an experimental model: light microscopy, electron microscopy and immunocytochemistry.

Two types of synthetic vascular grafts, Dacron Triaxial and Dacron Gelseal Triaxial, were implanted into both the common carotids of sheep. The animals were sacrificed 1, 2, 8, and 16 weeks after surgery. Multiple specimens, obtained from grafts and anastomoses, were studied by light microscopy, transmission and scanning electron microscopy. A parallel immunocytochemical analysis was performed on some specimens. Dacron Triaxial grafts failed to develop a complete neointimal coverage. Myofibroblasts and fibroblasts were the dominant cells in such synthetic graft. Moreover, focal areas of stripping, platelet deposition, and thrombosis were observed at 8 and 16 weeks. In contrast, a stable endothelial coverage developed on the Gelseal Triaxial grafts after 16 weeks.

Animals↗

Beam voltage effects in the study of embedded biological materials by secondary electron detectors.

Thin and semithin sections were extensively examined by the secondary electron (SE) detector in a conventional scanning electron microscope (SEM), and in a transmission electron microscope with a scanning attachment (STEM). Various parameters, in particular the beam voltage, were shown to affect the final SE image (SEI). As for SEM observation, a surface contrast was imaged at low primary electron (PE) voltages (0.6-2 kV), whereas a subsurface contrast predominated at higher energies (15-30 kV). In STEM, significant differences were not detected by varying the PE in the 20-100 kV range. Surface and subsurface information was simultaneously imaged even though the SEI were better resolved at the highest energy.

Carotid Arteries↗

Stromal cells in primary myelofibrosis: ultrastructural observations.

The bone marrows of five patients with primary myelofibrosis at different stages of the disease have been studied. In the myelofibrotic bone marrow, associated with "reticulum cells", two other cell types have been identified, namely fibroblast-like and myofibroblast-like reticulum cells, as well as a spectrum of transitional forms. Our findings suggest that reticulum cells may represent a reserve stromal cell pool (i.e. primitive reticulum cells) able to modulate themselves and to transform differently according to functional requirements. Some suggestions regarding the functional significance of fibroblast-like and myofibroblast-like reticulum cells in primary myelofibrosis are suggested.

Biopsy↗

T-cell prolymphocytic leukaemia: a clinical and immunological study.

2 cases of T-cell prolymphocytic leukaemia (T-PLL) were investigated for their reactivity with a series of monoclonal antibodies (MoAbs) as well as for the cytochemical expression and functional activity of the pathological cells. Both patients showed morphological (large cells with abundant cytoplasm and eccentric and irregularly shaped nucleus with large and prominent nucleoli) and clinical (high leucocyte count and splenomegaly) features typical of T-PLL. The cells from 1 patient expressed a helper/inducer phenotype (T4+, T8-) and were reactive with the anti-Tac (interleukin-2 receptor) MoAb, while the other case co-expressed both the T4 and the T8 antigens. The response to phytohaemagglutinin and the natural killer activity (assessed by 51chromium release) were significantly reduced in both cases, while the helper capacity, tested in a pokeweed mitogen-driven system, was maintained only in the 1st case. This latter case which expressed a more mature phenotype (T4+, T8-) responded well to chemotherapy.

Acid Phosphatase↗

Mechanically isolated hepatocytes are unsuitable to detect antibodies directed against plasma membrane determinants.

The ultrastructural morphology of mechanically and enzymatically isolated hepatocytes was compared. Plasma membrane integrity was evaluated by electron microscopy, both in the absence and in the presence of ruthenium red, which stains cell coat glycoproteins and makes it possible to identify even partially permeable cells, and by indirect immunofluorescence using experimental antibodies directed against epithelial submembranous components (prekeratin, actin). In order to evaluate the antigenic integrity of the hepatocellular plasma membrane after the isolation procedures, an immuno-electronmicroscopical technique (Staph. pA-colloidal gold) was applied using the IgG fraction from two liver-kidney microsomal antibody (LKM) positive sera, known to react with both plasma membranes and smooth endoplasmic reticulum (SER). Enzymatic procedure made it possible to isolate hepatocytes with a good preservation of both plasma membranes and subcellular organelles. Ruthenium red staining was confined to plasma membranes, thus indicating the preservation of both cell-coat glycoproteins and plasma membrane. Most cells were negative after exposure to experimental antibodies. After exposure to LKM positive sera colloidal gold was strictly confined to the plasma membrane. On the other hand, mechanically isolated hepatocytes showed wide interruptions of the plasma membrane and gross alterations of subcellular organelles. Most cells were stained by ruthenium red, thus confirming the plasma membrane permeability. In addition, a linear peripheral positivity was found in the vast majority of the cells tested with anti-prekeratin and anti-actin antibodies. LKM-colloidal gold complexes were found at the level of both residual plasma membrane and SER.(ABSTRACT TRUNCATED AT 250 WORDS)

Actins↗

Lineage relationship of chronic lymphocytic leukemia and hairy cell leukemia: studies with TPA.

The tumor promoting agent TPA (phorbol ester; 1.6 X 10(-8)M) was used to induce the differentiation in vitro of B-chronic lymphocytic leukemia (B-CLL) cells from 14 untreated patients. The uninduced phenotype was SIg+, Mrbc+, RFT-1+, RFA-4-, FMC7-. After 72 h incubation with TPA, B-CLL cells became RFA-4+, FMC7+ and lost the capability of Mrbc rosetting. Large proportions of the "induced" cells also showed morphological and ultrastructural changes, such as undulating membranes and bleblike protusions and became strongly positive for tartrate resistant acid phosphatase (TRAP+) and also contained cytoplasmic immunoglobulins. These features are very similar to the features of hairy cell leukemia (HCL). These observations confirm previous clinical findings that B-CLL and HCL are related disorders of the B lineage. The development of "hairy" features in induced B-CLL and in HCL seems to be a malignancy-associated feature because the Mrbc+ normal B cells (B-CLL-equivalent cells) isolated from tonsil also develop TRAP positivity but no membrane aberrations.

B-Lymphocytes↗

Ultrastructural aspects of human prostate in benign prostatic hyperplasia.

The ultrastructural morphology of the prostate in benign prostatic hypertrophy has been studied with particular attention focused on features of prostatic secretory processes and the formation of "corpora amylacea." Morphological and cytochemical comparisons of prostatic secretory vacuoles and multivesicular bodies have been carried out, and the secretory mechanisms of the prostate have been further defined by scanning electron microscopy.

Humans↗

Platelets in primary thrombocythemia: electron microscopic study.

The ultrastructural analysis of platelets from patients with Primary Thrombocythemia (PT) showed some anomalous features. Platelets having structural modifications of their organelles were observed and a decrease in the number of dense bodies was seen. Freeze-fracture analysis of the plasma membrane of platelets from PT patients showed a significant increase in the number of intramembranous particles (IMP).

Adult↗

Congenital dyserythropoietic anemia type II: serologic and morphologic family study.

A 29-year old woman with a history of chronic anemia, jaundice, and splenectomy was found to have congenital dyserythropoietic anemia type II (CDA II), based on the following: marrow erythroid hyperplasia, abundance of binucleated erythroblasts, electron microscopic evidence of double membranes lining the cell membrane, erythrocyte lysis in the acidified serum test, erythrocyte agglutination and lysis by anti-i antibodies. Three healthy relatives (mother, father, and brother) of the patient had marrow erythroid hyperplasia and a percentage of binucleated erythroblasts that was higher than normal. A structural alteration of the nuclear membrane was detected in some of their erythroblasts. Furthermore, the erythrocytes of the father and the brother were agglutinated by anti-i antibodies. This observation is consistent with a recessive mechanism of inheritance, and suggests that heterozygosity for CDA II can be somatically expressed at different levels.

Adult↗

Composite gastric carcinoma and precursor lesions with amphicrine features in chronic atrophic gastritis.

A composite carcinoma of the gastric body consisting of endocrine and mucous epithelial cells with interspersed amphicrine cells is reported together with ultrastructural and immunocytochemical documentation of endocrine and nonendocrine differentiation. The tumor was associated with hypergastrinemia related to chronic atrophic gastritis (achlorhydria) and with multiple proliferative lesions, such as intramucosal microcarcinoid (IMC) and endocrine cell proliferations of the micronodular and linear type, which are currently regarded as carcinoid precursor changes. Ultrastructurally, a composite architecture with amphicrine features was demonstrated in the primary tumor, IMC, and liver metastases. On the other hand, the endocrine cell proliferations exclusively contained gastrin and enterochromaffinlike cells. Immunostaining with antibodies to calcitonin documented a number of positive cells both in the primary and in the metastatic sites. This is the first report of mixed exocrine-endocrine-amphicrine components both in a metastasizing carcinoma and in its precursor lesions in a chronic hypergastrinemic state. Unlike previously reported lesions, the endocrine component was unexpectedly composed of calcitonin cells, which are not usually present in the gastric mucosa.

Aged↗

Filamentous inclusions in nonneoplastic and neoplastic pancreas: an ultrastructural and immunogold labeling study.

Filamentous inclusions (FI) are unusual, irregularly shaped cytoplasmic inclusions, which are mostly found in acinar cell carcinomas of the pancreas and are consequently thought to be an abnormal zymogen granule type. This study describes identical inclusions in acinar, centroacinar, and small duct epithelial cells from nonneoplastic pancreas, as well as those found in tumor cells from a mixed acinar-endocrine pancreatic carcinoma. An ultrastructural and immunogold labeling demonstration indicates that these inclusions are aggregates of intermediate filaments immunoreacting with the anti-cytokeratin AE1/AE3 mixture and with V9 clone anti-vimentin monoclonal antibodies. Their pleomorphic appearance, variable immunoreactivity, and frequent association with lipid droplets and secondary lysosomes, mostly of the angulate type, led to the hypothesis that the FI undergo a degenerative remodeling pathway similar to that proposed for hepatic Mallory bodies. A survey of the literature on FI and human tumors suggests that they are a variably expressed ultrastructural feature of tumor cells originating from exocrine cell-containing tissues, namely the pancreas and gastrointestinal tract.

Carcinoma, Acinar Cell↗

Inflammatory aspects in atherosclerotic abdominal aortic aneurysms. A case report.

An important inflammatory reaction was histologically observed in the wall of an abdominal aortic aneurysm in a 42 year-old patient suffering from multifocal atherosclerotic lesions. Histochemical and ultrastructural studies showed an active-stage wall lymphomonoplasmacellular component. The histological appearance and biochemical and microbiological studies excluded specific aortitis, a rheumatic disease or concomitant infectious processes. The morphological relationships between the atherosclerotic lesion and the infiltrate together with the immunophenotyping features of the infiltrate suggest the possibility of a close correlation between the inflammatory process and the atherosclerotic lesion of the wall.

Adult↗