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Biomedical subjects

G Garcia-Herdugo

Publications and source records attributed to G Garcia-Herdugo.

13 recordsLinked to original sources

Volatile anaesthetic halothane causes DNA damage in A549 lung cells.

The present study was performed to elucidate the extent of damage and the ability of lung epithelial cells to recover or to undergo apoptosis after in vitro treatment with the volatile anaesthetic halothane. The results obtained from the comet assay clearly show that halothane, applied at 3.0mM concentration, causes DNA and cell damage. Cells exhibited nuclear fragmentation and budding early after treatment and these events gradually increased during the next few days. The presence of a large number of mini-comets after single cell gel electrophoresis was found to represent apoptotic bodies with fragmented DNA. Our results demonstrate apoptosis-like changes after in vitro exposure of A549 cells to the volatile anaesthetic halothane. The majority of the affected cells did not recover and were directed to cell death.

Anesthetics, Inhalation↗

Ag-NOR proteins and rDNA transcriptional activity in plant cells.

In this work we used Allium cepa root meristem cells in actively growing conditions and under treatment with the protein synthesis inhibitors cycloheximide (CHM) and puromycin. Morphological and quantitative results indicate that these drugs induce dramatic alterations in nucleolar structure reflected by a decrease of nucleolar size, much more evident under treatment with CHM, and by segregation of its main components. Quantitative analysis shows a decrease in NOR-silver staining after treatment with CHM, whereas in cells treated with puromycin NOR-silver staining remains constant. Our results reveal a decrease in the Ag-NOR proteins under conditions of diminished cell activity, suggesting a direct relationship between the quantity of Ag-NOR proteins and transcriptional activity. Using two-dimensional gel electrophoresis and NOR-silver staining in gels, we have characterized some proteins corresponding to molecular weights of 28 and 31 KD and pI of approximately 5.2. After treatment with CHM, reactivity of these proteins against NOR-silver staining is diminished. By means of a morphological study, analysis of NOR-silver staining, and of anti-DNA and RNAse-gold labeling, we have tried an approach to the nucleolar organization in plant cells. Our results suggest that the fibrillar component shows a reticular distribution where fibrillar centers, as described in animal cells, are not distinguished.

Allium↗

Differential morphometric values induced in Golgi apparatus of higher plant cells by aldehyde and permanganate fixation.

In order to determine the best conditions to carry out quantitative ultrastructural studies in plant specimens, five different fixation techniques, including some of the most reported electron microscopy fixatives (glutaraldehyde-paraformaldehyde, osmium tetroxide, potassium permanganate), were assayed in onion root meristems to check their ability to induce morphometric changes in Golgi apparatus ultrastructure. Although the parameters evaluated showed in all cases the same tendency, values obtained after permanganate fixation were always higher than those found after aldehyde techniques (especially aldehyde-osmium). Aldehyde followed by osmium fixation appears as the most indicated fixation method when accurate quantitative ultrastructural studies are to be developed.

Aldehydes↗

Ultrastructural demonstration of lectin binding sites in the Golgi apparatus of rat epiphyseal chondrocytes.

Binding sites for wheat germ agglutinin (WGA), Dolichos biflorus agglutinin (DBA), Ricinus communis I agglutinin (RCA I) and Limax flavus agglutinin (LFA) have been ultrastructurally detected in rat epiphyseal chondrocytes by a post-embedding cytochemical technique using colloidal gold as marker. The four lectins labelled exclusively the Golgi apparatus of chondrocytes embedded in Lowicryl K4M resin by two different methods. WGA binding sites were localized in medial and trans cisternae as well as in immature secretory vesicles, whereas those for DBA were seen concentrated in cis and medial cisternae. Labelling with both RCA I and LFA lectins was distributed throughout all the cisternae of the Golgi stack, and the latter also in vesicles and tubules at the trans face. Neuraminidase pretreatment of the sections abolished LFA staining, decreased reaction with WGA and increased that with RCA I, while it did not affect DBA staining. After chondroitinase ABC treatment only the RCA I reaction was modified, revealing new binding sites in the trans Golgi face, secretory granules and extracellular matrix. These results indicate that the distribution of subcompartments in the Golgi apparatus of chondrocytes is different from that in cells secreting glycoproteins as major products.

Animals↗

Immunocytochemical and morphometric study of prolactin cells during amphibian morphogenesis.

The peroxidase-antiperoxidase (PAP) immunocytochemical technique and rabbit antihuman prolactin (PRL) antiserum were used to localize and identify PRL-producing cells in the pars distalis of Bufo calamita, Hyla meridionalis, Alytes cisternasii, Pelobates cultripes, and Rana perezi tadpoles at different stages of development as well as in 1-year-old post-metamorphic animals. This cell type was located throughout the gland in P. cultripes and R. perezi and in the caudal two-thirds in B. calamita, H. meridionalis, and A. cisternasii in premetamorphic animals. These distribution patterns do not show changes throughout development. Morphometry was used to evaluate the changes observed in pars distalis volume and stereological parameters of PRL immunoreactive cells during development. Pars distalis volume increased during the larval growth period and decreased throughout the metamorphic climax. PRL volume density and cellular area showed different patterns in the different species, although the correlation between these parameters suggests a period of great proliferative rate followed by changes in cellular size. The changes observed in PRL total volume suggest the existence of two phases in amphibian development: (i) a period of PRL storage during pre- and prometamorphosis and (ii) a period of release at the metamorphic climax.

Animals↗

Silver staining of the nucleolar organizer regions (NORs) in semithin Lowicryl sections.

The one-step silver technique was applied to semithin Lowicryl sections of root meristem cells of Allium cepa and a human tumor cell line (TG cells). In vegetal cells, after 5 min of staining reaction, the Ag-NOR proteins formed ring-shaped structures peripherally within the nucleolus. In animal cells silver granules were distributed over the entire nucleolus. The specificity of the staining reaction was increased by incubation of the sections in NH4Cl and Schiff's reagent prior to Ag-NOR silver staining.

Allium↗

Lectin binding patterns in amphibian epidermis.

A battery of 6 different horseradish peroxidase conjugated lectins has been employed for structural localization of glycoconjugates in amphibian epidermis. Lens culinaris (LCA) lectin stained the basal membrane and gave no significant reaction on the epidermal layers. Canavlia ensiformis (Con A) and Griffonia simplicifolia II (GS II) lectins bound the keratinocyte cytoplasm and the basal membrane as well. Ulex europaeus I (UEA I) lectin had only reactivity with flask cells. Griffonia simplifolia I (GS I) and Glycine max (SBA) lectins preferentially bound the cell membranes of keratinocytes, being the intensity of the staining gradually increasing from the stratum spinosum to the stratum granulosum. These results show that UEA I, GS I, and SBA are good markers to distinguish different cell types and the degree of keratinocytes differentiation.

Animals↗

A morphometric study of subcellular responses in frog pituitary corticotropic cells to constant environmental conditions.

Ultrastructural morphometry (point-counting method) was used to evaluate stereological parameters of rough endoplasmic reticulum (RER), Golgi complex, secretory granules, mitochondria and lysosomes of Rana ridibunda pituitary corticotropic (ACTH) cells in animals subjected to constant environmental conditions (CEC) in relation to temperature and photoperiod. During this experiment, corticotropic cells are particularly sensitive to the effects of changes in environmental conditions in such a way that they course through a phase of RER and Golgi complex hypertrophy after 7 days on CEC and a phase of enhancement in the amount of secretory granules (the 11th day). After 15 days on CEC, the cells seemingly recuperate almost all the control evaluate parameters. These results strongly suggest that, in frog kept in captivity, ACTH cells play an important role in the acclimation process to new constant environmental conditions.

Adrenocorticotropic Hormone↗

Subcellular responses in frog pituitary gonadotrophic cells to constant environmental conditions: a morphometric study.

An ultrastructural study of the pars distalis of the pituitary in Rana ridibunda specimens subjected to constant environmental conditions revealed significant structural differences which indicate a short-time increase in the activity of gonadotrophic cells. Planimetry was used for morphometric and stereological analysis of the rough endoplasmic reticulum, Golgi complex, secretory granules and lysosomes. The presence of a vesicular endoplasmic reticulum throughout the cytoplasm of gonadotrophic cells was observed in the pituitary gland of the intact frog. After 7 days of constant environmental conditions, there was an increase in the volume density of the endoplasmic reticulum vesicles, while on the 11th day there was a slight reversal in the vesicle content that was complete the 15th day. In the hypertrophy of the endoplasmic reticulum, the larger vesicles (bigger than 0,52 micron2) play a more important role than the smaller ones (similar to the endoplasmic reticulum observed in control cells). However, the small vacuoles were also hypertrophied in relation to the control. Degranulation was also observed after 7 days of constant environmental conditions. On the 15th days, there was an increase in the numerical density of secretory granules and lysosomes compared to the control while their volume density was similar to the control.

Animals↗

An electron microscopic study of stellate cells and cavities in the pars distalis of frog pituitary.

Stellate cells in the pars distalis of adult Rana ridibunda were observed electron microscopically under normal and experimental conditions (TRH injection). The stellate cells have lengthy processes extending into the intercellular spaces between the secretory cells and scanty cytoplasm surrounding the nucleus. Occasional desmosomes link stellate cells to adjacent secretory cells. In the pars distalis of animals injected with thyrotropic-releasing hormone (TRH), the stellate cells form large cavities (2-6 mum) filled with heterogeneous material. Their cytoplasm contains well-developed Golgi complexes and some lysosomes; these are the principal morphological alterations as compared to those observed in control animals. It is suggested that stellate cells could play an active role in addition to providing a structural framework for the pars distalis.

Animals↗