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M Fuenzalida

Publications and source records attributed to M Fuenzalida.

15 recordsLinked to original sources

Antiangiogenic effect of betamethasone on the chick cam stimulated by TA3 tumor supernatant.

Tumor growth is the result of combined cell proliferation overwhelming cell death and neoangiogenesis. This report shows CAM angiogenesis promoted by TA3 tumor supernatant with or without low dosis of betamethasone (Minimal antiangiogenic concentration: beta-MAAC). Methylcellulose discs instilled with 10 microliters of beta-MAAC (0.08 microgram/ml), 10 microliters of tumor supernatant (TA3ts), 5 microliters beta-MAAC + 5 microliters TA3ts, and 10 microliters of PBS as control were implanted in host chick eggs. On day 12, the grafts were removed, photographed and fixed. Sections were stained in parallel, one and three with hematoxylin-eosin, and section two by the Tunel method. The number of vessels was evaluated in a microscopic field of the CAM (2250 micron 2). The results show that beta-MAAC produced a significant inhibition of neovascularization in comparison to that observed in controls (P < 0.0025; Student t-Test). Discs instilled with TA3ts produced an intense stimulation of angiogenesis in contrast, when discs were instilled with 5 microliters of beta-MAAC + 5 microliters of TA3ts the angiogenesis was significantly inhibited (P < 0.001). The results show that effective antiangiogenic doses of betamethasone are in the range of 10(-7) M, (probably a genomic mediated action) and that this effect of low concentration may have clinical applications.

Allantois↗

Histochemical detection of sugar residues in lizard teeth (Liolaemus gravenhorsti): a lectin-binding study.

The structural diversity of the many oligosaccharide chains of surface glycoconjugates renders them likely candidates for modulators of cell-interactions, cellular movements, differentiation, and cellular recognition. A selection of different lectins was used to investigate the appearance of cellular distribution and changes in sugar residues during tooth development in the polyphyodont lizard, Liolaemus gravenhorsti. Lectins from three groups were used: (1) N-acetylgalactosamine specificity: BS-1, PNA, RCA-120; (2) N-acetylglucosamine specificity: ECA; and (3) fucose specificity: UEA 1 and LTA.. Digital images were processed using Scion Image. Grayscale graphics in each image were obtained. The lectins used showed a strong, wide distribution of the L-fucose and N-acetylgalactosamine at the cell surface and in the cytoplasm of multinucleate odontoclast cell, while mononuclear odontoclast cells showed no binding, suggesting some roles that the residues sugar might play in the resorption of dentine or with multinucleation of odontoclast after the attachment to the dentine surface in this polyphyodont species. Further studies must be planned to determine the specific identities of these glycoconjugates,and to elucidate the roles played by these sugar residues in the complex processes related to odontogenesis in polyphyodont species.

Acetylgalactosamine↗

Microscopic and histochemical study of odontoclasts in physiologic resorption of teeth of the polyphyodont lizard, Liolaemus gravenhorsti.

Using tartrate-resistant acid phosphatase (TRAP), we examined the cytodifferentiation of odontoclast cells in resorbing areas of dental tissues during the replacement of teeth in a polyphyodont lizard, Liolaemus gravenhorsti. We also report, by means of Lectin-HRP histochemistry, the distribution pattern of some specific sugar residues of TRAPase-positive cells. For detection of TRAPase activity, the azo dye-coupling technique was used. Lectin binding sites were demonstrated by means of specific HRP-lectins. The process of tooth resorption was divided into four stages: 1) preresorption-the wall of the dental pulp is covered with an odontoblast layer, and no TRAP-positive cells are in the dental pulp; 2) early resorption-TRAP-positive multinucleate odontoclasts are present on the dental wall, but the rest of the pulp surface is still covered with an odontoblast layer; 3) later resorption-the entire surface of the pulp chamber is lined with multinucleate odontoclasts; and 4) final resorption-the tooth has been totally resorbed. Odontoclasts are usually detached from the resorbed surface, and show signs of degeneration. Of the six lectins used, PNA, ECA, and UEA-1 bind to multinucleated but not mononuclear cells. All the remaining lectins, BS-1, RCA(120), and LTA showed no binding to any cells of the teeth. The significance of saccharidic moieties such as acetyl-galactosamine, acetyl-glucosamine, and fucose sugar residues is difficult to ascertain. Perhaps these oligosaccharides might be borne on molecules associated with odontoclastic resorption or associated with multinucleation of odontoclasts after attachment to the dentine surface.

Acetylgalactosamine↗

Effect of cholinergic agonists on muscular tonus of the lizard small intestine and esophagus.

The underlying mechanisms of acetylcholine-induced intestinal relaxation in the lizard Liolaemus tenuis tenuis are still unknown. By using a classical model of intestinal recording of isometric contraction and relaxation in conjunction with specific pharmacological tools, this article studies the possible influence of EDRF/NO and nicotinic ganglionar receptors on the Ach-induced relaxation in an effort to elucidate the probable mechanisms involved in ACh effect. It was observed that the relaxation of the lizard intestine elicited by ACh (10(-7) - 4 x 10(-4) M) was not affected by hexametonium (5 x 10(-4) M) or tetrodotoxin (10(-6) M). Nicotine (10(-7) to 10(-4) M) induced relaxation was significantly antagonized by hexametonium; however, it was not influenced by tetrodotoxin. These results allow us to discard a neuronal pathway in cholinergic-induced relaxation, suggesting a more direct cholinergic effect on the smooth muscle, perhaps mediated by an unknown substance released by some specialized tissue. N-nitro-L-arginine, used to block NO-synthase and NO production, induced no changes in ACh-induced relaxation. Methylene blue, a soluble guanylate cyclase inhibitor, induced no changes in ACh-induced relaxation. These results allow us to discard a probable role of EDRF/nitric oxide in the ACh-induced relaxation of lizard small intestine, providing evidence that this mechanism could be different from that reported in other species.

Animals↗

Ethanol effect on the chick embryo ossification: a macroscopic and microscopic study.

This study attempts to analyze anomalies in avian embryos induced macroscopically and microscopically when exposed to ethanol (EtOH) during the first stages of development. Fertilized chicken eggs were employed in this study. The eggs were incubated at 37.8 degrees C. Some of the eggs were treated on day 0 with EtOH (20%, 40% and 60%) by instillation in the air sac. The control group was instilled with 0.1 ml of NaCl at 0.9%. Other eggs were treated on the 4th post-incubation day, employing the same methodology. The embryos in both groups were removed from the eggs on the 11th incubation day and examined using a dissecting binocular microscope. After macroscopic analysis, the samples obtained were fixed in 10% formol, photographed and processed according to common histological techniques and the Picrosirius method. Embryos treated with EtOH demonstrated a significant weight decrease. Microscopic analysis by means of the Picrosirius method revealed that the intra-membranous ossification process presents less development, and therefore there was less type I collagen in trabecular bone in the embryos post-exposure to EtOH with respect to the control.

Abnormalities, Drug-Induced↗

Detection of sugar residues in rabbit embryo teeth with lectin-horseradish peroxidase conjugate: II. A light microscopal study.

The cellular distribution and changes of sugar residues during tooth development in embryos of the rabbit Oryctolagus cuniculus were investigated by using horseradish peroxidase-conjugated lectins (lectin-HRP). The lectins SBA, ECA, and LTA show no binding to any region of the dental cap and bell stages, whereas BS-1 and UEA-1 bind to dental cells at both stages. Appropriate control studies confirmed the specificity of the binding of the lectins. At cap stage, the lectins BS-1 and UEA-1 show moderate binding to the (pre)-ameloblast and (pre)-odontoblast cells. These results suggest that the acetylgalactosamine and alpha-L-fucose residues present in (pre)-ameloblasts and (pre)-odontoblasts, respectively, are common to determined but relatively undifferentiated cells capable of forming matrices of hard tissues. Since the odontoblast and ameloblast express dentin and enamel, respectively, it can be speculated that the abundance of these residues in these cells might be associated with the maintenance of the capacity of the cells to produce such matrices. At the bell stage, the odontoblasts display considerable amounts of alpha-L-fucose, whereas alpha-L-fucose is poorly localized in ameloblasts. However, ameloblasts contain significant quantities of N-acetylgalactosamine, whereas only a diffuse positivity for this carbohydrate is apparent in odontoblasts. The marked changes of the glycosylation pattern of these glycoconjugates might indicate that they play a role during the cell-to-cell interaction and might also be involved in the odontoblastic and ameloblastic functional activity. Such a possibility is entirely speculative until specific in vitro experiments are conducted.

Animals↗

Light microscopic detection of sugar residues in rabbit embryo teeth with lectin-horseradish peroxidase conjugates.

We investigated the binding of five HRP-conjugated lectins to rabbit tooth germs at the cap and late bell stages of development. The results revealed some changes in the glycosylation patterns of the glycoconjugates. Sugar residues, such as alpha-D-mannose, methyl-D-glucose, N-acetylglucosamine, beta-D-galactosamine, D-galactose, and sialic acid, were detectable in some components of the tooth germs. The most conspicuous developmental change was increased binding of Con A and WGA. These lectins showed, at the cap stage, moderate binding to the (pre)-ameloblasts and (pre)-odontoblasts. With further development to the late bell stage, but prior to the achievement of well-defined morphological-functional characteristics, the odontoblasts and ameloblasts displayed considerable amounts of alpha-D-mannose, alpha-D-glucose as well as beta-D-acetylglucosamine and sialic acid. Appropriate control studies confirmed the specificity of the binding of the lectins. Two lectins (DBA and PNA) with known specificity for N-acetylgalactosamine groups were bound by the basement membranes in tooth germs at the cap stage. A third lectin (RCA) with the same specificity did not produce any detectable staining in the same material. Further studies must be planned to determine the specific functions and significance of lectin-HRP-binding glycoconjugates in odontogenesis.

Animals↗

Behavior of rabbit dental tissues in heterospecific association with embryonic quail ectoderm.

The behavior of dental tissues from the rabbit, Oryctolagus cuniculus, in association with epithelium from the quail, Coturnix coturnix japonica, has been examined. Adult and embryo rabbits were employed in this study. Dental papillae from teeth at the cap stage from rabbit embryos and dental pulp from adult rabbits were isolated surgically and recombined with skin ectoderm from 72-hour-old quail embryos. The recombined tissues were cultured for 48 hours on semi-solid medium and subsequently removed and placed on chorio-allantoic membranes of 7-day-old chick embryos. Control cultures (dental pulp, dental papillae, and quail ectoderm) showed regression, atrophy, or differentiation according to the phenotype of the tissue. After 8 days in explant culture, heterologous recombinants composed of dental papillae and flank skin ectoderm from quail embryos developed differentiated chimeric tooth structures. It was unclear whether or not enamel was being secreted. The fact that the interactions between the enamel epithelium and the dental papillae are reciprocal is well known. The differentiation of odontoblasts can only occur in the presence of an enamel organ. Thus, the quail epithelium must have been induced to become an enamel organ, the lack of enamel proteins notwithstanding. Apical pulp and root pulp from adult rabbits plus quail ectoderm showed a high degree of regression and atrophy. At around 15 days of gestation, the rabbit dental papillae at the cap stage have already acquired odontogenic potential. By contrast, under the same experimental conditions, the dental pulp from continuous-growth teeth from adult rabbits did not show odontogenic potential.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Odontogenesis and amelogenesis in interacting lizard-quail tissue combinations.

In this study we examined the possible inductive role of the dental papilla from polyphyodont lizard tooth germs. Flank skin sheets of quail ectoderm enzymatically separated from dermal tissue were recombined with lizard tooth papillae and placed on semisolid medium and cultured for 2 days. Subsequently, the recombinants were removed and placed on the chorioallantoic membrane of chick hosts and incubated for 6 days. After this period of 8 days in explant, control tissues differentiated according to their own phenotypes. Lizard dental papilla alone differentiated as fibroblasts. Quail flank skin ectoderm differentiated into epithelial sheets. Intact lizard tooth buds developed into teeth with dentine and incipient enamel. In the best experimental recombinants, advanced and relatively well-constructed teeth were observed, with clear indications of hard tissue deposition in association with quail epithelium. The results show that mesenchyme of the adult lizard dental papilla and embryonic quail ectoderm of heterotopic origin are capable of carrying out the complex sequence of morphogenetic interactions involved in normal odontogenesis.

Amelogenesis↗

Ultrastructural aspects of dental tissues and their behavior in xenoplastic association (lizard quail).

Ultrastructural characteristics of tooth buds of the polyphyodont adult lizards Liolaemus tenuis and Liolaemus gravenhorsti have been elucidated. Xenoplastic combinations of lizard whole tooth buds and neural crest cells from embryos of the quail Coturnix coturnix japonica have been cultured in vitro. Mesenchymal cells (preodontoblasts) of lizard teeth early develop filopodia that contact the basal lamina. Fragments of quail neural crest isolated by dissection were recombined with isolated lizard tooth buds and cultured for 84 hours in dishes kept in an incubator at 37.8 degrees C in air. Some identifiable quail cells in these recombinants developed a cytoplasmic extension like that of an odontoblastic process. These results suggest that lizard tooth rudiments already determined for tooth development produce some non-species specific transmissible constituents which are capable of inducing quail cranial neural crest cells to express certain dental characteristics (odontoblastogenesis) not expressed in their normal development in vivo.

Animals↗

In vitro differentiation of tooth buds from embryos and adult lizards (L. gravenhorsti): an ultrastructural comparison.

It is well established that the capacity for teeth to differentiate "in vitro" depends upon: a) the age of the embryonic rudiments at the time of excision and b) the number of cells within each tissue type which are capable of differentiating into organ culture. This paper studies ultrastructural aspects of tooth buds grown in vitro from lizard embryos and compares these characteristics with those observed in dental germs grown in situ in older lizard embryos. Moreover, we report the self-differentiation in vitro dental tissues from adult lizard and compare this phenomenon with the main features of a morphogenetic field. Our results suggest that approximately in the first third of gestation in L. gravenhorsti the dental buds has already acquired the capacity for self-differentiation in vitro. The ultrastructural observations show that there are no significant differences between odontoblasts and ameloblasts in situ and in vitro. The tooth from "adult lizards," isolated by combined microsurgical and enzymatic procedure and cultured in semisolid-liquid medium were also able to differentiate teeth. This phenomenon implies that self-differentiation is not rigidly determined, and that in these animals the tooth tissues represents a continuous morphogenetic field throughout the animal's life. This property is intrinsic, resides in the isolated tooth tissues, and is relatively independent of external factors. In addition, these studies indicate that the chick chorio-allantoic membrane and the semisolid-liquid culture medium supply the majority of the factors required for development of these tissues.

Ameloblasts↗