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Biomedical subjects

F Beltrame

Publications and source records attributed to F Beltrame.

At least 19 recordsLinked to original sources
PubMed1997

Studies on the structure of sperm heads of Eledone cirrhosa by means of CLSM linked to bioimage-oriented devices.

We have used a confocal laser scanning optical microscope imaging device and a bioimage-oriented workstation equipped for augmented reality to study the helical sperm head of the octopus Eledone cirrhosa. This approach allows us to study different complex organisational motifs due to the spatial arrangement of linear helical structures. We consider this helical specimen an enlarged copy of one of the most important biostructures governing cell functioning such as chromatin-DNA. Moreover, this very same sample is made of highly compacted chromatin that can be studied at higher resolution, i.e., by means of scanning force microscopy. Fluorescence optical sectioning has been used to enter the spatial organisation. Three-dimensional images of single, twisted, and folded fibers are shown.

Animals↗
PubMed1996

Time-variant analysis of organelle and vesicle movement during phagocytosis in Paramecium primaurelia by means of fluorescence confocal laser scanning microscopy.

Vital fluorescent dyes (FITC-albumin, Texas Red-albumin, and acridine orange) were used together with a confocal laser scanning optical microscope (CLSM) to display and analyze formation, movement, and fusion of vesicles during the phagocytosis of Paramecium primaurelia, in the x-y-z-t space. By immobilizing living cells pulsed with a food vacuole marker at successive times after chasing in unlabeled medium, the intracellular movement of food vacuoles from their formation at the cytostome to their egestion at the cytoproct was visualized, and food vacuoles were selected in a specific digestion stage. Small pinocytic vesicles are shown to evaginate from the vacuoles and move in the cytoplasm. These vesicles are transported toward the cytopharynx where they enlarge the membrane of the nascent food vacuoles or fuse with stage II food vacuoles, when the vacuoles of stage II increase their size, changing from an acidic to an alkaline status. A multimodal analysis of confocal fluorescence images and the false-color technique were used to visualize vesicle movement vs. time. Starting from three images of the same cell at succeeding time points, a composite image was generated by associating with each originally acquired image a different color corresponding to each sampling point in time. The composite image shows that vesicles move away from the food vacuole in a scattered manner exhibiting changes in direction.

Animals↗
PubMed1995

Image analysis of lysosomal activity during the early clonal life of Paramecium primaurelia.

Acid phosphatase activity was measured in individual cells by determining their optical densities through a scanning confocal laser microscope. The naphthol AS-TR (3-hydroxy-2-naphtoic acid 4'-chloro-2'-methylanilide) phosphate-hexazotized para-rosanilin method was used to visualise the acid phosphatase content in the light microscope. Evidence was obtained that the amount of enzyme varied in exponential growth phase cells as the fission age increased. By comparing the acid phosphatase activity with the rate of food vacuole formation, it appeared that the amount of enzyme inside the cells decreased in early clonal life, whereas the rate of food uptake increased. It was assumed that the reduction of acid phosphatase content could lead to a more extended life of vacuoles and to a decreased membrane recycling rate. In turn, the reduced supply of membrane available for food vacuole formation could partly be responsible for the decrease of the food uptake rate observed after the initial increase.

Acid Phosphatase↗
PubMed1995

Pressure control ventilation and minitracheotomy in treating severe flail chest trauma.

OBJECTIVE: To evaluate pressure control ventilation (PCV) delivered through a minitracheotomy in treating severe flail chest trauma. DESIGN: Case report. SETTING: Intensive care unit of a trauma center. PATIENT: A 34-year-old woman affected by flail chest trauma and acute respiratory failure, who was initially treated with tracheal intubation to obtain internal pneumatic stabilization. The patient failed extubation and noninvasive mask treatment (pressure support ventilation plus PEEP) due to poor chestwall mechanics. INTERVENTIONS: Minitracheotomy was performed and ventilation was achieved with high levels of inspiratory pressure (PCV or assisted PCV) to overcome the resistance of the cannula (Mini-Trach II, Portex, ID 4 mm). Esophageal and carinal pressures were monitored. Ventilatory treatment was always performed with the full cooperation of the patient; the patient's glottic function was always intact. The patient was successfully treated with pressure control ventilation delivered through the Mini-Trach. After 7 days of PCV, the patient was switched to assisted PCV. On the 20th day after admission, she was weaned from mechanical ventilation. CONCLUSIONS: We conclude that a suitable gas exchange and pneumatic stabilization in a flail chest condition can be achieved using minitracheostomic ventilation. At the same time, this treatment could reduce some side effects of traditional tracheal intubation.

Adult↗
PubMed1995

Acid phosphatase activity in mating type I and mating type II cell lines of Paramecium primaurelia.

The cellular acid phosphatase content, a marker enzyme for lysosomal activity, in Paramecium primaurelia mating type I and mating type II cells was determined by optical laser scanning microscopy. The naphthol AS-TR phosphatase-hexazotized pararosaniline method was used to visualize acid phosphatase activity by the light microscopy. Cell lines of both mating types were tested during culture life, from the early log phase to the death phase. The amount of acid phosphatase was higher in mating type II than in mating type I until the onset of the stationary phase, and then the values reversed. Indeed, during the log phase of growth, mating type II cells formed a higher number of food vacuoles, so that, by taking up a higher amount of bacteria, they sooner became deprived of food. It is suggested that, by lacking nutrients, their synthesis activities and acid phosphatase content were reduced as compared with mating type I cells.

Acid Phosphatase↗
PubMed1993

Confocal microscopy and cellular bioinformatics.

An exhaustive description of most biological data requires, besides a set of analytic information, an eidetic representation of the data itself. In this paper, two topics are presented. The first one is confocal microscopy, an advanced technique to produce multidimensional cellular and subcellular structures images. The second one is a software application, named BIOCELL, based on a relational database management system, developed to provide the biologist with an integrated tool to handle simultaneously cell biology linguistic and eidetic information.

Biotechnology↗
PubMed1992

Dystrophin at the plasma membrane of human muscle fibers shows a costameric localization.

We studied the distribution of dystrophin at the sarcolemma of normal human muscle fibers using high resolution immunofluorescence and confocal laser scanning optical microscopy (CLSOM). We found that the dystrophin lattice is organized at the muscle plasma membrane in an array of thick bands interconnected by a finer network. The bands encircle the muscle fiber perpendicular to the long axis of the fiber and they matched the sites of attachment of the sarcomeres to the plasma membrane. Dystrophin co-localized with vinculin, and dystrophin and vinculin co-localized with alpha-actinin at the region of the I-band. Dystrophin may be one of the proteins involved in the linkage of the sarcomeres to the extracellular matrix.

Actinin↗
PubMed1984

Automated analysis of living cells through the quantitative use of automated phase contrast microscopy.

A simplified theory of image formation in phase contrast microscopy is presented. It is shown that the phase shift induced in light (related to the refractive index) by the observed object can be reconstructed, point by point, from the phase-contrast digitally sampled image through an appropriate algorithm. This allows one to make quantitative observations on unstained, living cells.

Animals↗
PubMed1983

Higher-order structure of chromatin from resting cells. II. High-resolution computer analysis of native chromatin fibres and freeze-etching of nuclei from rat liver cells.

Non-destructive electron microscopy of native chromatin from rat liver nuclei reveals that the 30 nm fibre is formed of four 11 nm nucleofilaments, arranged in a coiled-coil (or rope-like) conformation. At low ionic strength, native fibres show an alternating pattern of compact and unwound regions. Freeze-etching experiments carried out on the same nuclei are compatible with the existence of periodic attachments of the fibres to the nuclear envelope near the pores in a regular, drapery-like fashion. For the first time, computer image analysis has been applied to electron micrographs of giant chromatin fibres and a few essential geometrical parameters characterizing the conformation of the higher-order structures have been determined. No significant difference has been found between calf thymus and rat liver chromatin.

Animals↗
PubMed1980

The quinternary chromatin-DNA structure. Three-dimensional reconstruction and functional significance.

Nuclear DNA-space images from Feulgen-stained HeLa cells synchronized at 1, 3, 5, 8, 12, 15, and 18 h following mitosis are digitized and their densitometric-geometric patterns are analyzed by means of a Quantimet 720-D image analyzer on line with a PDP11/40 computer. Frequency distributions of picture point optical densities for the phases and subphases as seen in nuclear images show that DNA packing changes are evident by means of ordinary optical microscopy. Radii of gyration of the images, and optical density profiles and distributions for several squashes of similar cells reveal that in particular instances chromatin DNA is distributed mostly towards the periphery, and usually with high circular isotropy. Cross power spectra of individual scan lines suggest that existence of higher order "quinternary" periodic structure for chromatin that modulates during the cell cycle. Three-dimensional reconstruction 2- micrometer sections of intact, Feulgen-stained mammalian tumor tissue show stainable material only toward the nuclear perimeter and not in the center (compatible with the evidence that initial thymidine incorporation in HeLa cells is generally at the nuclear border). Densitometric properties of reconstructed interphase chromatin-DNA bodies are highly coupled with similar properties of the whole nucleus, showing that a more condensed nucleus is always accompanied by a more condensed interphase chromatin DNA. The effect of micrococcal nuclease digestion on the digitized nuclear images is also presented. All the above data are then discussed in terms of a quinternary chromatin-DNA structure and its modulation during the cell cycle.

Cell Cycle↗
PubMed1979

Morphometric analysis of B2cAMP induced reverse transformation in synchronized CHO cells.

Synchronized tranformed and reverse-transformed (by 10(-3) M B2cAMP) CHO-K1 cells, growing adherent to plastic, are characterized by means of geometric and densitometric parameters at the level of both the entire cell and of the nuclei at various time intervals after selective miotic detachment. Transformed and reverse-transformed cells triple-stained with Feulgen, Napthol Yellow S, and periodic acid-Schiff appeared very similar in terms of integrated optical density (IOD), related to either polysaccharides, protein, or DNA amount. On the other hand, a shift from a polygonal to a spindle-shaped morphology is a accompanied by a significant decrease in both form factor and average optical density (AOD) of intact cell and nuclei, which are the most conspicuous measured changes caused by B2cAMP, in addition to a lengthening of the cell cycle duration. In both control and treated cells, important and parallel cell-cycle-dependent modulations of geometric and densitometric parameters are also observed, for both the cytoplasmic (i.e., cell morphometry) and DNA space (i e., nuclear morphometry). Specifically, the modulation in nulear morphometry during G1, S, G2, and M phases confirms previous findings on synchronized HeLa cells. The optical density threshold-dependence of geometric parameters shows that, while becoming fusiform, the cytoplasm of reverse-transformed cells had a particularly low optical density precisely in the polar area. Utilization of such an approach in the development of an objective morphological classification of all cell lines grown as monolayers "in vitro" is also discussed.

Animals↗