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Biomedical subjects

S Squarzoni

Publications and source records attributed to S Squarzoni.

8 recordsLinked to original sources

Morphological evidence of function-related localization of phospholipids in the cell nucleus.

The evidence accumulated in recent years on the presence of phospholipids inside the interphase nucleus needs a precise localization of the nuclear sites of accumulation, transport and degradation of these molecules. A very useful approach for monitoring the fine localization of nuclear phospholipids is represented by a recently developed technique using gold-conjugated phospholipases. In fact, in addition to the phospholipids organized in bilayers in the membrane, this technique identifies amorphous lipoprotein complexes present in different cell areas as well as in the nucleus. In this way and using sample preparation systems which reduce lipid removal and translocation, such as cryofixation, cryosectioning, embedding in hydrophylic resins and cryofracturing, we have analyzed the subnuclear localization of phospholipids in different experimental conditions. The results indicate that: in interphase the nuclear phospholipids are localized mainly in the interchromatin spaces and in the nucleolar domain; the observed co-localization of phospholipids and ribonucleoproteins suggests that phospholipids are involved in the mechanism of transport and release of the transcripts; the demonstrated release of ribonucleoproteins after phospholipase digestion suggests that phospholipids mediate the binding between ribonucleoproteins and the nuclear matrix; significant changes of the phospholipid localization occur in the different phases of the cell cycle or in the course of induced cell differentiation.

Animals

Localization of dystrophin COOH-terminal domain by the fracture-label technique.

The precise localization of dystrophin in the skeletal muscle cell should contribute to a better understanding of the yet unclear functional role of this protein, both in normal and in Duchenne muscular dystrophy. Immunocytochemical studies did not give conclusive results on the localization of dystrophin with respect to the sarcolemma and to the cytoskeletal components. To improve the reliability of the electron microscopic immunocytochemical localization of dystrophin, a mAb against the COOH-terminus of the molecule has been used in association with the fracture-label technique, which, causing a partition of the membrane in protoplasmic and exoplasmic halves, allows a more precise dystrophin localization. The results obtained indicate that dystrophin is associated with the protoplasmic half of the plasmalemma, and the observation that it does not randomly follow the partition of the membrane is consistent with a stable association with the cytoskeleton.

Cell Membrane

Intranuclear localization of phospholipids by ultrastructural cytochemistry.

The presence of phospholipids within the interphase nucleus and in isolated chromatin, previously demonstrated by analytical biochemical methods, has been only rarely documented by cytochemical procedures, especially at the ultrastructural level. By means of a gold-conjugated phospholipase technique, we investigated the fine localization of endogenous phospholipids in the different nuclear domains in rat pancreas and in cell cultures. To reduce possible removal or displacement of phospholipids, different specimen preparation procedures such as cryofixation, cryosectioning, and freeze-fracturing were utilized. Apart from slight differences in efficiency among these methods, phospholipids have been cytochemically identified in the same nuclear domains: the interchromatin granules and fibers and the dense fibrillar component of the nucleolus. These results suggest that the phospholipids are an actual nuclear component, not randomly distributed in the nucleoplasm but mainly localized in the nuclear domains involved in the synthesis, maturation, and transport of ribonucleoproteins.

Animals

Image analysis of the chromatin organization in the nuclear domains of freeze fractured hepatocytes and lymphocytes.

The complex organization of the interphase nucleus can be analyzed, by way of thin sectioning and also freeze-fracture. This approach has previously been utilized in association with image analysis to quantitatively describe the organization of isolated rat liver nuclei and nuclear matrices. The main nuclear domains which, in section, present marked differences due to their electron-density, can be identified in replicas with more complex procedures, based on the quantitative evaluation of the number of particles per unit area and mainly by using image analysis. A quantitative analysis of the nuclear substructures has been performed by way of image analysis on in situ nuclei of freeze-fractured cells presenting marked differences in the heterochromatin quantity, such as hepatocytes and lymphocytes. The replicated nuclear particles have been classified according to their diameter and the obtained histograms have been quantitatively evaluated. The nuclear domains, heterochromatin, interchromatin, nucleolus, present characteristic ratios among the three main classes of particles; that is, ribonucleoproteins, solenoid filaments and solenoid fibre aggregates. The typical patterns of the nuclear domains can be further stressed by selecting a single class of particles and by examining its topographic localization. While interchromatin and nucleolar domains present a similar quantitative pattern in hepatocytes and lymphocytes, the heterochromatin of lymphocytes contains a significative higher percentage of solenoid aggregates than that of hepatocytes.

Animals

Morphometric study of chromatin pattern in freeze-fractured rat liver nuclei during malignancy evolution.

Computerized image analysis was performed on freeze-fractured nuclei isolated from normal rat liver and rat liver with diethylnitrosamine induced hepatocarcinomas. Chromatin particles belonging to heterochromatin and interchromatin nuclear regions were measured, distributed into histograms and statistically processed. The ratio between nucleosome filaments (5-10 nm range diameter) and solenoid fibres (25-35 nm range) and Tukey's fitting line were calculated. The N/S ratio shows an inverse trend in treated nuclear samples with respect to the controls indicating that the malignancy progression is accompanied by characteristic derangement of the chromatin arrangement which can be evaluated by morphometric analysis.

Animals

Considerations on ceramic prosthesis explants.

The authors report data obtained for 11 cases of explants of prostheses having ceramic joint surfaces. The wear phenomena observed in 9 prostheses, including 1 case with breakage of the ceramic head, was moderate: 20-22,000 debris per mu 2. In 2 cases the wear was massive, with a concentration of debris in the peri-implant tissues 5-10 times greater (100,000-218,000 debris per mu 2). An explanation for the trigger mechanism of wear phenomena is proposed: the cases of massive wear are related to loosening of the acetabulum. A histological examination of the bone tissue surrounding a stem covered with alumina (Al2O3) carried out in three cases allowed us to observe an area of demineralization measuring approximately 500 mu of tissue in contact with the ceramic coating. The osteomalacia could be caused by the diffusion of alumina ions.

Bone Cements