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Biomedical subjects

G Pasquinelli

Publications and source records attributed to G Pasquinelli.

At least 91 records · Page 5Linked 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↗

Brenner tumor of the ovary: a correlative histologic, histochemical, immunohistochemical, and ultrastructural investigation.

The histologic, histochemical, immunohistochemical, and ultrastructural features of Brenner tumor (BT) were studied. BT was compared with transitional bladder cells, and close similarities between the two tissues were identified. Abundant glycogen in all cellular layers, an alcianophilic/sialomucinic surface mucous coat, and argyrophilic cells characterized both BT and bladder epithelium. Immunohistochemically, chromogranin and neuron-specific enolase reactivity was observed in all cases examined. An additional relevant finding was the presence of serotonin-storing cells in both BT and urothelium. Moreover, carcinoembryonic antigen, epithelial membrane antigen, and keratin reaction were found in BT and urothelium, indicating an additional antigenic similarity. Additionally, malignant Brenner tumor was ultrastructurally found to share many common features with the bladder tissue. The distinct histochemical, ultrastructural, and antigenic pattern of BT, primarily of the transitional type, is emphasized.

Brenner Tumor↗

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↗

Scanning electron microscopy application in clinical research.

Our personal experience on the application of scanning electron microscopy in cardiology, gastroenterology and ophthalmology is reviewed. SEM has not yet significantly contributed to myocardium pathology. However, in the near future, SEM could be a reliable technique to complete the information available from other sources. As to atherosclerosis, SEM allowed us to improve our knowledge of the early stages of the disease; some pathological features, not always detected by conventional morphological examinations, can be documented. An important contribution to gastrointestinal pathology was made by SEM investigations both in the staging of some important diseases (i.e., coeliac disease, peptic ulcer, Crohn's disease, ulcerative colitis) and in the follow-up of mucosal changes during therapy. In the ophthalmological field, SEM provided three-dimensional new information to clinicians, who are familiar with the biomicroscopic images. Our experience in hematology is still limited. However, in the last few years SEM joined to immunocytochemistry allowed us to characterize cell populations in several blood diseases. Some procedures of particular interest in the management of human bioptic specimens are stressed in order to get to a complete correlative microscopy. We conclude that continuous and simultaneous correlations have to be carried out between SEM and other methods and instruments available for morphological investigation.

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↗

S.E.M. study I: Gastric and duodenal lesions induced by non-steroidal anti-inflammatory drugs (aspirin, piroxicam) in man.

A scanning electron microscopy (SEM) study was performed on the gastric and duodenal mucosa of 14 patients treated with Aspirin or Piroxicam. Six patients with normal mucosal morphology were treated with either 1.5 g/day of Aspirin (3 patients) or 20 mg/day or Piroxicam (3 patients) for one month. In addition, 8 rheumatic patients were treated with a similar dose of Aspirin (4 patients) or Piroxicam (4 patients) for at least 4 months. SEM was used to evaluate the following parameters: mucosal structure, cellular exfoliation and anisocytosis, alteration of the microvilli (blebs). The mucosa showed various aspects of alterations ranging from minimal changes (microvillar alteration) to a completely subverted structure both in the stomach and in the duodenum. Mucosal changes, both on the edge of the macroscopic lesions and at a distance from these, were visible with SEM even when endoscopy was normal. Piroxicam appeared slightly less damaging than Aspirin, and no difference was observed between short-term and long-term treated patients, either with Aspirin or with Piroxicam.

Adult↗

S.E.M. study II: Protective effect of ranitidine against gastric and duodenal lesions induced by non-steroidal anti-inflammatory drugs.

The effect of Ranitidine on the gastric and duodenal lesions induced by Aspirin or Piroxicam was studied by scanning electron microscopy (SEM). Twelve patients were entered into the study. The first group consisted of 6 patients with normal basic morphology, of which 3 were treated with 1.5 g of Aspirin and Ranitidine 300 mg/day, and 3 with 20 mg of Piroxicam and Ranitidine 300 mg/day, for one month. The second group was composed of 6 rheumatic patients, 3 of which took 1.5 g of Aspirin and the other 3 received 20 mg of Piroxicam per day for at least four months. Ranitidine (300 mg/day) was then administered concomitantly for one month to all these 6 patients. Overall, Ranitidine appeared to have a positive effect in preventing the development of gastroduodenal lesions induced by these anti-inflammatory drugs in the first group of patients. No differences were observed in the ability of Ranitidine to promote the healing of lesions induced by chronic treatment. Ranitidine, however, appeared to be more effective in preventing lesions induced by Aspirin rather than those induced by Piroxicam.

Adult↗

A new approach for studying semithin sections of human pathological material: intermicroscopic correlation between light microscopy and scanning electron microscopy.

In order to obtain useful and complete information on the study of pathological material, we observed by scanning electron microscopy (SEM) the same semithin sections observed by light microscopy (LM). For this purpose, the specimen must have, at the same time, chromatic and electron dense characteristics. We thus developed different specimen preparation methods, subjecting the semithin sections to specific polychromatic staining with high atomic number (Z) elements, to monochromatic staining followed by routine contrasting with uranyl acetate and lead citrate, and to specific cytochemical and immunocytochemical procedures. The specimens were examined in sequence by LM, by SEM equipped with secondary electron, backscattered electron, transmitted electron detectors and by scanning transmission electron microscopy (S(T)EM).

Actins↗

Clinical applications of scanning electron microscopy in gastrointestinal diseases.

We considered the role of scanning electron microscopy (SEM) in clinical investigation of different gastrointestinal diseases. The following clinical applications of SEM may be suggested on the basis of our original data and those reported in literature: in peptic ulcer: assessment of the completeness of healing, by observing the mucosal surface architecture of the scars; identification of mucosal changes, namely enterocytic surface membrane alterations, predictive of recurrence; in coeliac disease: early assessment of the response to gluten-free diet and follow-up of the patients by staging the process of mucosal repair in cerebriform, intermediate and villous patterns; in ulcerative and Crohn's colitis: enhancement of the diagnostic sensitivity of perendoscopic biopsy, by detecting differences in surface structure of mucosa surrounding ulcers in both diseases. This is subverted in ulcerative colitis and preserved in Crohn's colitis. Finally the complementary role of SEM in relation to endoscopy and light microscopy is emphasized.

Adolescent↗

Use of secondary electron detectors for compositional studies on embedded biological material.

In a scanning electron microscope (SEM) the secondary electron (SE) detector collects SE from various sources (SE-I, SE-II, SE-III) as well as backscattered electrons (BSE). SE-II and SE-III are generated by BSE and therefore provide compositional data. In order to collect the compositional electrons in the SE and BSE signals, we employed specimens with little or no surface morphology. Thin and semithin sections of biological embedded material were utilized, conventionally stained and/or subjected to cytochemical or immunocytochemical procedures. The specimens were observed in a SEM equipped with SE detector (SED), BSE detector (BED) and transmission electron (TE) detector (TED). We drew the following conclusions: a) images by SED of thin sections give the same information as images by TED; b) images by SED of semithin sections provide complementary information with respect to those obtained in BSE and TE mode. Some parameters affecting the intensity and the quality of the signal collected were also considered. Finally we observed the same semithin sections in a transmission electron microscope (TEM) with a SE attachment. Images obtained by SED, BED and TED were fully diversified.

Carotid Arteries↗

Fresh and cryopreserved arterial homografts: immunological and clinical results.

INTRODUCTION: This prospective study defined the immunological and clinical results after fresh and cryopreserved arterial homograft replacement due to graft infection. MATERIALS AND METHODS: Thirty patients who underwent ABO-compatible homograft transplantation were studied for anti-human leukocyte antigen (HLA): antibody production and CD3- and CD4- versus CD8-positive lymphocyte subsets. Nine patients (30%) received immunosuppressive treatment with cyclosporine (1 to 3 mg/kg/d). Immunological studies were performed preoperatively, and early (1, 3, 7 days) and late (1, 3, 6, 12, 24, 36, 48 months) during follow-up. Abdominal computed tomography scans were performed postoperatively at 1, 6, 12, 24, 36, and 48 months of follow-up. RESULTS: Preoperatively, antibodies were not detected. Postoperatively, a progressive increase in percent panel reactive antibodies was observed in all patients 1 month after the transplant. There were no difference between fresh and cryopreserved homografts. The antibody response among patients treated with cyclosporine was less pronounced and delayed. Recipient antibodies were directed against donor-specific antigens. During the immediate postoperative period (1, 3, 7 days) there was a slight increase in CD3- and CD4-positive T lymphocytes and a concomitant decrease in the CD8 subset. Later, CD3 and CD4 progressively decreased and the CD8 set increased. Clinically, no patients had signs of recurrent infection upon late follow-up. Four patients died (13%), but only one death was homograft-related (rupture of the graft). At 2-year follow-up, two patients showed stenotic lesions due to chronic rejection. Clinically, no differences were noted between fresh and cryopreserved homografts, or between patients treated with or without cyclosporine. CONCLUSIONS: Fresh and cryopreserved arterial homografts are immunogenic; they induce a strong anti-HLA antibody response, similar to chronic rejection.

ABO Blood-Group System↗

Desmoplastic small round-cell tumor: a case report on the large cell variant with immunohistochemical, ultrastructural, and molecular genetic analysis.

examination. The patient died 10 months after surgery. Histologically, the tumor was composed of predominantly large epithelioid cells with foci of anaplasia mimicking metastatic carcinoma. Immunohistochemically, the tumor cells stained with anti-cytokeratin, EMA, desmin, and NSE antisera. Electron microscopy showed secretory lumina, desmosomes, cell processes with microtubules and electron-dense granules, and focal whorls of intermediate filaments. Reverse transcriptase-polymerase chain reaction performed on paraffin block-retrieved tissue demonstrated the EWS/WT-1 fusion transcript characteristic of the t(11;22)(p13;q12). This case illustrates a less common histological pattern of DSRCT, i.e., diffuse large cells, thus supporting the view that this tumor presents a wider morphological spectrum than that previously recognized.

Abdominal Neoplasms↗

Myofibrosarcoma of the upper jawbones: a clinicopathologic and ultrastructural study of two cases.

Two problematic spindle cell sarcomas involving upper jawbones in two adult male patients have been studied by histology, immunohistochemistry, and transmission electron microscopy, and respectively graded as low-grade malignancy and high-grade malignancy. While any single methodological study did not allow confident classification of them into one or other of the classical categories of spindle cell sarcomas (fibrosarcoma versus leiomyosarcoma), the overall contribution from all three methodologies ultimately allowed them to be categorized as sarcomas with myofibroblastic differentiation. Histologically, both tumors had morphological features of an amalgama between neoplastic fibroblasts and smooth muscle cells. Immunohistochemically, both tumors expressed reactivity only for muscle specific actin and alpha smooth muscle actin, in addition to vimentin. Ultrastructurally, both tumors, while showing fibroblast-like cytoplasmic features, had a spurious and imperfectly organized cell surface defying convincing classification into any of specific categories (i.e., both appeared in terms of ultrastructure as poorly differentiated sarcoma, the former with low level of smooth muscle differentiation and possibly the presence of some fibronexus component, the latter with no smooth muscle differentiation but with possible evidence of very rare fibronectin fibril). Therefore, on balance, the most tenable diagnosis seemed to us that of a myofibrosarcoma in both cases. This work is presented considering the fact that myofibrosarcoma currently represents a topical theme of debate, and that this is the first report in medical literature concerning with myofibrosarcomas of the head and neck area in adults.

Actins↗