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Biomedical subjects

G Delfino

Publications and source records attributed to G Delfino.

At least 19 recordsLinked to original sources

Serous cutaneous glands of the Pacific tree-frog Hyla regilla (Anura, Hylidae): patterns of secretory release induced by nor-epinephrine.

The serous (poison) cutaneous glands of the Pacific tree-frog Hyla regilla were induced to release their product by 10(-3)M nor-epinephrine stimulation. After discharge structural and ultrastructural features of the cutaneous glands involved in release were observed. Furthermore, the discharged product, consisting of discrete, secretory granules, was collected and processed for transmission electron microscope analysis. As indicated by patterns found in the myoepithelium encircling the syncytial secretory unit, gland discharge is caused by contraction of the peripheral myocytes. Muscle cell compression dramatically affects the syncytium and results in degenerative changes, including expulsion of the secretory unit nuclei. Therefore, the structural collapse in depleted glands has been ascribed to the mechanical activity performed by the myoepithelium during discharge, rather than cytoplasm involution described in conventional, holocrine glands. TEM investigation revealed that the secretory granules collected after discharge maintain their peculiar traits: they consist of recurrent patterns of thin subunits, acquired during serous maturation and provided with remarkable structural stability.

Amphibian Venoms↗

Development of serous cutaneous glands in Scinax nasica (Anura, Hylidae): patterns of poison biosynthesis and maturation in comparison with larval glands in specimens of other families.

We examined the development of serous (poison) cutaneous glands in larval and juvenile Scinax nasica (Hylidae) at the ultrastructural level. We describe the biosynthesis and maturation of the cutaneous poison in comparison with the corresponding processes in representatives of Discoglossidae, Leptodactylidae, Pelobatidae and Pipidae. Serous biosynthesis in S. nasica starts in discrete adenoblasts and continues in the syncytial secretory unit. Biosynthetic processes involve rough endoplasmic reticulum and the Golgi apparatus, that releases membrane-bounded material, varying from fine grained to flocculent. During the post-Golgian secretory phase, this material undergoes initial maturation, and two products are formed: dense granules and larger vesicles holding a thin substance that will later be structured into a three-dimensional, honeycomb-like net. Both the secretory granules and vesicles change into glomerular-like aggregates of bowed, rod-shaped subunits (modules). In adult frogs, formation of dense granules is bypassed. The modular granule substructure seems to be related to the merocrine release of small amounts of poison, involved in regulating skin homeostasis. Comparison with maturational changes in larval glands of species representing four anuran families discloses similar patterns in the Leptodactylidae, but production of opaque homogeneous granules occurs in the Discoglossidae, clear vesicles in the Pelobatidae and aggregates of dense bars in the Pipidae.

Animals↗

Ultrastructural patterns of secretory activity in poison cutaneous glands of larval and juvenile Dendrobates auratus (Amphibia, Anura).

A transmission electron-microscope study has been performed on larval and juvenile skin of the Central American arrow-frog Dendrobates auratus to investigate early secretory processes and maturational changes in the serous (poison) glands. Poison biosynthesis involves the endoplasmic reticulum (both smooth and rough types), as well as Golgi stacks which release early serous product as secretory vesicles (or pre-granules). These vesicles contain fine-grained material, along with single electron-opaque bodies, spheroidal in shape, that accompany the grained product throughout its post-Gogian, maturational change. The first steps of this process involve condensation and lead to the formation of secretory granules with a glomerular-like substructure, resulting from a thick, random aggregation of rods (secretory granule subunits). Advanced maturational activity causes the loss of peculiar granule substructure: the dense bodies split into fragments, whereas the thick glomerular arrangement becomes looser, until the secretory product changes into a dispersed material. This ultrastructural study revealed biosynthesis and maturation processes in close sequence, suggesting the poison of D. auratus contains proteins and/or peptides as well as lipophilic compounds. Molecules of both these classes are known to perform several roles relevant to survival strategies in extant anurans. Furthermore, the ephemeral granules with a glomerular-like substructure detected in tadpoles and froglets exhibit the complex patterns of mature poisons in adult specimens of other anurans: Hylidae and related families. This agrees with current trends in the taxonomy of these advanced frogs and underlines the pertinence of an ontogenetic approach in investigating anuran phylogenesis.

Amphibians↗

Secretory granule-cytoplasm relationships in serous glands of anurans: ultrastructural evidence and possible functional role.

A survey covering the serous (granular) cutaneous glands in several anuran families from the Old and New Worlds (Bombinatoridae, Discoglossidae, Ranidae, Hylidae, Pseudidae and Leptodactylidae) has revealed consistent patterns of complex interactions between the syncytial secretory unit and serous deposits (granules). These relationships involve outgrowths from the syncytial cytoplasm encircling the granules and complex invaginations of the perigranular compartment (halo) into the syncytium. The outgrowths are branched, cytoplasm processes resembling ramified microvilli, or can be larger, dome-like to cylindrical structures. Despite their different features and origins, all these structures are efficient devices for amplifying the cytoplasmic surfaces round the granules, so improving exchange between the secretory syncytium and serous product. These complex secretory granule-cytoplasm interactions affect the product released from the Golgi apparatus and are consistent with the hypothesis of prolonged serous maturation following the initial phase of biosynthesis. Post-Golgian maturation modifies the secretory material on a centripetal gradient, causing condensation and, possibly, the transfer of component molecules from and/or to the cytoplasm.

Animals↗

Ectal mandibular gland in Polistes dominulus (Christ) (Hymenoptera, vespidae): ultrastructural modifications over the secretory cycle.

An ultrastructural study was carried out on the secretory activity of the ectal mandibular gland in the wasp Polistes dominulus (foundress and worker females as well as males). Secretory activity in foundresses proceeds slowly during hibernation and early spring, becoming prominent in late spring and then falling sharply during the summer. This sequential pattern of ultrastructural modifications follows a functional, annual cycle. However, by comparing the subcellular changes in the gland with colonial development, it appears that secretory activity fits in with the specie's social cycle rather than merely following the seasons. The highest levels of secretory activity correspond to the early, critical breeding phases, while activity slows down with an increase in colony protection, based on both primary (passive) and secondary (active) defenses, with the emergence of the workers. These correlations suggest that the ectal mandibular gland secretory product in P. dominulus is involved in chemical nest defense.

Animals↗

Secretory polymorphism and serous cutaneous gland heterogeneity in Bufo granulosus (Amphibia, Anura).

Three types of secretory products (a, b and c) in the poison glands of the Argentine toad Bufo granulosus have been detected under light microscope. The type a secretory product consists of granules of homogeneous density, type b of vesicles with a translucent compartment and type c of granules of varying density. Subsequent transmission electron microscope analysis disclosed obvious similarities in the secretory pathways of type a and c granules; the differences detected under light microscope are due to the functional phases observed. On the contrary, production of type b secretory vesicles involves a distinctive pathway. Therefore, two classes of glands (I and II) have been identified. Glands of the first class are typical of bufonid toads and produce granules provided with repeating substructure; glands of the second class, which manufacture a lucent product, are unusual in the family Bufonidae. Ultrastructural differences, consistent with the two gland classes, have also been described in the myoepithelia. The myocytes ensheathing class I secretory units possess striking cytoskeletal specializations, whereas those of class II glands are rich in sarcoplasmic reticulum. The distinctive ultrastructural traits detected in these myoepithelial cells have been compared with the results of previous studies on the dimorphic serous glands of Bombina. Findings point to the use of pharmacological treatment on the skin of anurans with different classes of serous glands to elicit differential secretory discharge.

Animals↗

Serous gland dimorphism in the skin of Melanophryniscus stelzneri (Anura: Bufonidae).

Two serous gland types (I and II) in the skin of the Argentine toad Melanophryniscus stelzneri were discovered using light and electron microscopy. Glands of the two types differ in several traits: features of the products (both mature and immature), organelles involved in biosynthesis, and paths of serous maturation. No consistent differences, however, were detected between the myoepithelial sheaths encircling the secretory units. Type I glands manufacture vesicles containing a single dense body with a repeating substructure and conform to the fundamental secretory line of bufonid skin, a secretory line involved in biosynthesis of steroids. Type II glands synthesize granules of varying densities and seem to belong to a line of glands that secrete proteinaceous products. The occurrence of the two serous gland types in Melanophryniscus stelzneri is discussed in a comparison with current literature on the morphofunctional characteristics of anuran poison glands, which perform both regulative and defensive roles. It is suggested that di- or polymorphism in serous glands is an adaptive trait that allows differential release of active molecules on the body surface.

Animals↗

Serous cutaneous glands in Phyllomedusa hypochondrialis (Anura, Hylidae): secretory patterns during ontogenesis.

Three syncytial gland types (Ia, Ib, and II ) have been described in the skin of larval, juvenile and adult Phyllomedusa hypochondrialis, which share the ultrastructural traits common to the serous secretory units of anuran skin, although each manufactures a peculiar product. Type la secretion consists of dense granules provided with a peculiar substructure, type Ib of vesicles holding a lucent material, type II of lipid deposits. None of the developmental stages investigated showed intermediate features between any of the three cutaneous products, which accumulate in the syncytial cytoplasms of the secretory units following different biosynthetic pathways, consistent with each gland type. These findings confirm previous results on adult specimens of P. hypochondrialis and P. sauvagei and stress the polymorphism of the serous glands in the genus Phyllomedusa. This morphological variability reflects the wide adaptive flexibility of serous glands in anurans.

Animals↗

Cutaneous venom of Bombina variegata pachypus (Amphibia, Anura): effects on the growth of the human HL 60 cell line.

The effects of Bombina variegata cutaneous venom (Bvv) on eukaryotic cell growth has been assessed employing the human leukaemic cell line HL 60, by liquid and agar semisolid cultures and 51Cr release assay. HL 60 cells growth is impaired by Bvv in a dose-dependent fashion in both culture systems. The arrest of proliferation requires a contact time lower than 3 min and it is not reversed by washing and culturing the cells in a Bvv-free medium. Similarly, an extremely short exposure time is needed to determine maximum 51Cr release. Neither the agar medium nor the fetal calf serum interact with Bvv effects, which, according to the above findings, must be regarded as cytolytic in nature. In both liquid and the agar-semisolid culture Bvv cytolytic activity half life is about 8 hr. The cytolytic properties of Bvv are thought to be part of the chemical defence system of amphibian skin.

Amphibian Venoms↗

A bactericidal protein in Bombina variegata pachypus skin venom.

The skin venom of the yellow bellied toad Bombina variegata pachypus has an antimicrobial activity which seems to be correlated to the presence of a 6700 mol. wt polypeptide. This polypeptide was purified by electroelution from SDS-urea-polyacrylamide gels and characterized for its antimicrobial activity. A bactericidal action was detected at concentrations with little or no cytolytic effect. The determination of the Minimal Inhibitory Concentration showed that there was activity against gram positive and gram negative bacteria and also against yeasts. The skin secretions of three other anuran species (Bufo viridis, Hyla arborea and Discoglossus pictus) were examined for the presence of antimicrobial activities. Only the Hyla arborea secretion exhibited antimicrobial properties. A small amount of a 6700 mol. wt polypeptide was detected among the Hyla secreted products.

Amphibian Venoms↗

Cytochemical study of larval skin in Amphibia: occurrence of ruthenium red positive cells in cutaneous gland anlagen.

During a cytochemical investigation into the development of cutaneous gland buds in amphibian larvae (both anuran and urodele), ruthenium red (RR) positive cells were found in the anlagen. These cells, with diffuse deposits of the polycation dye in their cytoplasms, were first detected on the lateral surface of still intraepidermal gland buds. Later, during segregation of anlagen from the epidermis, the reactive cells were followed whilst they converged towards the apex of the gland nest. These findings of a remarkable penetration of RR through intact plasma membranes are discussed in the light of the staining characteristics of the polycation dye. Furthermore, the mechanism regulating morphogenesis of amphibian cutaneous glands is briefly compared with the morphogenetic patterns of large exocrine glands in higher vertebrates. It is suggested that during fixation RR penetrates morphogenetically active cells, following the same pathways as calcium under physiological conditions. Calcium inflow correlates with the occurrence of thin, possibly contractile, filaments in these cells and also agrees with cell migration during morphogenesis.

Animals↗

A low molecular weight protein with antimicrobial activity in the cutaneous 'venom' of the yellow-bellied toad (Bombina variegata pachypus).

The cutaneous 'venom' was collected from dorsal skin fragments of the yellow-bellied toad Bombina variegata pachypus by means of stimulation with noradrenaline. Light and electron microscope observations gave evidence that the 'venom' corresponds to the secretory products of both serous gland types (i.e. with small or large granules) characteristic of this genus, which had discharged their contents upon stimulation. The serous 'venom', when tested for antimicrobial activity, inhibited the growth of several bacterial strains. Heat treatment, dialysis, protease digestion and SDS-PAGE electrophoresis showed that the antimicrobial activity was thermostable and associated with a low molecular weight protein. This protein was purified and homogeneity determined by CM-cellulose chromatography and SDS-PAGE electrophoresis. The purified protein has a molecular weight of 6700, displays antibacterial properties and appears different from the antimicrobially active peptides previously isolated from the 'venom' of the toad.

Animals↗

Renal selectivity properties towards endogenous albumin in minimal change nephropathy.

It is well accepted that the molecular charge and conformation of serum proteins are major determinants of their glomerular filtration, but few studies characterizing the molecular features of circulating proteins in renal diseases are currently available. In 11 children affected by minimal change nephropathy (MCN) we determined the electrical charge and the fluorescence quantum yield of Tyrosine (Tyr) and Tryptophan (Trp) (taken as index of conformation) of serum and urinary albumin before and after steroid-induced remission of proteinuria. In all proteinuric children at the onset of the disease, urinary albumin was formed by one band with an isoelectric point (pI) of 4.7 (pI of the native protein), and by numerous other, less anionic bands with pIs between 4.8 and 5.5 accounting for about 50% of the total amount of this protein. The normalization of proteinuria which followed steroid therapy was characterized by the disappearance in urines of the less anionic fraction and by the appearance of numerous isoforms with a pI still more anionic (pI less than 4.7) than normal. At the same time, in the proteinuric phase, the fluorescence quantum yield of Trp of urinary albumin was markedly quenched, returning to near normal levels after steroid-induced remission of proteinuria. These data indicate that in MCN the charge-dependent renal selectivity properties are partially maintained and that the less anionic isoforms of albumin are a main component of urinary albumin. Together with the electrical charge, the conformation of albumin as a major determinant of its urinary excretion in MCN must also be considered.

Adolescent↗

Reactivity with diazonium salts of albumin from micro and macroalbuminuric diabetic patients.

A recent theory of the pathogenesis of diabetic microalbuminuria points to an involvement of glycated albumin, which has been demonstrated as being able to fluetrate the renal filter. The chemical characterization of urinary albumin, initially performed on the electrical charge and conformation of the protein has now been extended to the affinity properties for specific chemical probes. In this context, urinary albumin from Albustix-negative diabetic patients was found to be highly reactive towards diazonium salts (a dye specific for pyrrole rings) while the same protein purified from macroproteinuric diabetics showed no difference in reactivity towards diazonium salts compared to serum or normal albumin. These data indicate that, beside being highly anionic and conformationally deranged, urinary albumin in conditions of normal renal selectivity contains pyrrole structures. The reasons for considering this reactivity as an indirect sign of rearranged structure are presented here.

Adult↗