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Ricardo Boland - One of the best experts on this subject based on the ideXlab platform.

  • in vitro effects of 1α 25 oh 2d3 glycosides from solbone a Solanum glaucophyllum leaves extract herbonis ag compared to synthetic 1α 25 oh 2d3 on myogenesis
    Steroids, 2016
    Co-Authors: Valeria Gili, Ricardo Boland, Verónica González Pardo, Ana Carolina Ronda, Pablo De Genaro, Heini Bachmann, Ana Russo De Boland
    Abstract:

    The presence of glycoside derivatives of 1α,25(OH)2D3 endows plants to gradual release of the free bioactive form of 1α,25(OH)2D3 from its glycoconjugates by endogenous animal tissue glycosidases. This results in increased half-life of the hormone in blood when purified plant fractions are administered for therapeutic purposes. In this work, we evaluated the role 1α,25(OH)2D3-glycosides enriched natural product (Solbone A) from Solanum glaucophyllum leaf extract compared with synthetic 1α,25(OH)2D3 on myogenic differentiation in C2C12 myoblasts. For these, differentiation markers and myogenic parameters were studied in C2C12 myoblasts. Results showed that Solbone A, likewise the synthetic hormone, increased creatine kinase activity at day 2 after differentiation induction (60%, p<0.05). Solbone A and synthetic 1α,25(OH)2D3 increased vitamin D3 receptor protein expression at 10nM (50% and 30%, respectively) and the transcription factor myogenin (80%, p<0.05). However, tropomyosin expression was not affected by both compounds. In addition, myosin heavy chain (MHC) protein expression was increased 30% at day 2 of differentiation. Solbone A or synthetic 1α,25(OH)2D3 had no effects on myogenin nor MHC cell localization. Cellular mass increased with myogenesis progression, being Solbone A more effective than synthetic 1α,25(OH)2D3. Finally, Solbone A, as well as synthetic 1α,25(OH)2D3, augmented the index fusion of cultured muscle fibers. In conclusion, these results demonstrated that Solbone A exhibit at least equal or greater effects on early myoblast differentiation as synthetic hormone, suggesting that plant glycosides could be an effective, accessible and cheaper substitute for synthetic 1α,25(OH)2D3 to promote muscle growth.

  • In vitro effects of 1α,25(OH)2D3-glycosides from Solbone A (Solanum glaucophyllum leaves extract; Herbonis AG) compared to synthetic 1α,25(OH)2D3 on myogenesis
    Steroids, 2016
    Co-Authors: Valeria Gili, Ricardo Boland, Verónica González Pardo, Ana Carolina Ronda, Pablo De Genaro, Heini Bachmann, Ana Russo De Boland
    Abstract:

    The presence of glycoside derivatives of 1α,25(OH)2D3 endows plants to gradual release of the free bioactive form of 1α,25(OH)2D3 from its glycoconjugates by endogenous animal tissue glycosidases. This results in increased half-life of the hormone in blood when purified plant fractions are administered for therapeutic purposes. In this work, we evaluated the role 1α,25(OH)2D3-glycosides enriched natural product (Solbone A) from Solanum glaucophyllum leaf extract compared with synthetic 1α,25(OH)2D3 on myogenic differentiation in C2C12 myoblasts. For these, differentiation markers and myogenic parameters were studied in C2C12 myoblasts. Results showed that Solbone A, likewise the synthetic hormone, increased creatine kinase activity at day 2 after differentiation induction (60%, p

  • presence of vitamin d3 receptor vdr like proteins in Solanum glaucophyllum
    Physiologia Plantarum, 2006
    Co-Authors: Lorena Magdalena Milanesi, Ricardo Boland
    Abstract:

    Fil: Milanesi, Lorena Magdalena. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Bahia Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biologia, Bioquimica y Farmacia; Argentina

  • Presence of estrogen receptor (ER)-like proteins and endogenous ligands for ER in solanaceae
    Plant Science, 2004
    Co-Authors: Lorena Magdalena Milanesi, Ricardo Boland
    Abstract:

    Abstract The synthesis of steroid hormones in different plant species and the possibility that these molecules could regulate cell growth, tissue differentiation and germination has been reported. However, the mechanism of action of these endogenous steroids in plant cells is still poorly understood. In the present work, binding experiments with [ 3 H]17β-estradiol in the presence and absence of an excess of unlabeled 17β-estradiol showed that Solanum glaucophyllum and Lycopersicon esculentum organs contain estrogen-binding sites. Scatchard analysis detected saturable [ 3 H]17β-estradiol-binding sites ( K d ∼6.6 nM; B max ∼1140 fmol/mg protein) in S. glaucophyllum callus tissue. Estrogen-like compounds were detected in lipid extracts from S. glaucophyllum and L. esculentum . Moreover, the lipid fraction was able to compete with [ 3 H]17β-estradiol for binding to estrogen receptor (ER) from breast cancer MCF-7 cells as well as with estrogen-binding sites present in both plant species. Western blot analysis with monoclonal antibodies against different domains of the ER α, detected a ∼67 kDa band in various organs of both plant species. These proteins were able to bind estradiol in ligand blot assays using 17β-estradiol macromolecular derivatives as ligands. Western and ligand blot experiments of subcellular fractions from callus tissues of S. glaucophyllum showed that the ER-like protein of ∼67 kDa was most concentrated in the nuclear fraction. Reactive bands of lower molecular weight were also detected. Altogether these results provide evidence about the existence of estrogen-binding proteins and endogenous ligands in Solanaceae.

  • Vitamin D compounds in plants
    Plant Science, 2003
    Co-Authors: Ricardo Boland, Alejandro Carlos Curino, Mario Skliar, Lorena Magdalena Milanesi
    Abstract:

    Abstract An appreciable number (15) of flowering plants, specially within the Solanaceae family, have been shown to contain vitamin D 3 and its hydroxylated derivatives, including 1α,25(OH) 2 -vitamin D 3 [1α,25(OH)D 3 ], a pluripotent hormone in animals. These secosteroids have also been detected in members of the Cucurbitaceae, Fabaceae and Poaceae families. On the basis of recent cladistic analysis it is possible to predict that the synthesis of these compounds is a characteristic of the Angiosperms. Highly specific and sensitive bioassays and analytical procedures of high resolution are now available which may allow confirmation of this hypothesis. Solanum glaucophyllum is the species which accumulates 1α,25(OH)D 3 to the greatest extent. The metabolite is distributed among different tissues of the plant as a free steroid or glycoside derivatives. The presence of 7-dehydrocholesterol, vitamin D 3 , 25(OH)D 3 and 1α,25(OH) 2 D 3 in S. glaucophyllum and other species has been unequivocally demonstrated. Moreover, recent studies with radioactive precursors, protein immunoblot and RNA hybridization analysis suggest the presence of specific hydroxylases for vitamin D 3 and 25(OH)D 3 structurally related to the corresponding enzymes in vertebrates. It appears then that plants possess a similar synthetic route to 1α,25(OH) 2 D 3 as in animals. However, of unique biological importance, evidence obtained with S. glaucophyllum and Nicotiana glauca supports the operation of a non-photolytic reaction of vitamin D 3 synthesis. Both intermediates and enzymes of the vitamin D 3 pathway can be detected in cell and tissue cultures thus affording a convenient experimental model for studies on its molecular characterization and regulation. This is also endowed with biotechnological significance. Vitamin D 3 compounds may play a function in Angiosperms. Studies in vitro have shown that, like mitogenic plant hormones, they stimulate root growth and differentiation through activation of the Ca 2+ messenger system. There are vitamin D 3 and 1α,25(OH) 2 D 3 binding proteins in plants which exhibit characteristics suggesting a regulatory function. It remains to be established whether these binders act as true steroid hormone receptors.

Lorena Magdalena Milanesi - One of the best experts on this subject based on the ideXlab platform.

  • presence of vitamin d3 receptor vdr like proteins in Solanum glaucophyllum
    Physiologia Plantarum, 2006
    Co-Authors: Lorena Magdalena Milanesi, Ricardo Boland
    Abstract:

    Fil: Milanesi, Lorena Magdalena. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Bahia Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biologia, Bioquimica y Farmacia; Argentina

  • Presence of estrogen receptor (ER)-like proteins and endogenous ligands for ER in solanaceae
    Plant Science, 2004
    Co-Authors: Lorena Magdalena Milanesi, Ricardo Boland
    Abstract:

    Abstract The synthesis of steroid hormones in different plant species and the possibility that these molecules could regulate cell growth, tissue differentiation and germination has been reported. However, the mechanism of action of these endogenous steroids in plant cells is still poorly understood. In the present work, binding experiments with [ 3 H]17β-estradiol in the presence and absence of an excess of unlabeled 17β-estradiol showed that Solanum glaucophyllum and Lycopersicon esculentum organs contain estrogen-binding sites. Scatchard analysis detected saturable [ 3 H]17β-estradiol-binding sites ( K d ∼6.6 nM; B max ∼1140 fmol/mg protein) in S. glaucophyllum callus tissue. Estrogen-like compounds were detected in lipid extracts from S. glaucophyllum and L. esculentum . Moreover, the lipid fraction was able to compete with [ 3 H]17β-estradiol for binding to estrogen receptor (ER) from breast cancer MCF-7 cells as well as with estrogen-binding sites present in both plant species. Western blot analysis with monoclonal antibodies against different domains of the ER α, detected a ∼67 kDa band in various organs of both plant species. These proteins were able to bind estradiol in ligand blot assays using 17β-estradiol macromolecular derivatives as ligands. Western and ligand blot experiments of subcellular fractions from callus tissues of S. glaucophyllum showed that the ER-like protein of ∼67 kDa was most concentrated in the nuclear fraction. Reactive bands of lower molecular weight were also detected. Altogether these results provide evidence about the existence of estrogen-binding proteins and endogenous ligands in Solanaceae.

  • Vitamin D compounds in plants
    Plant Science, 2003
    Co-Authors: Ricardo Boland, Alejandro Carlos Curino, Mario Skliar, Lorena Magdalena Milanesi
    Abstract:

    Abstract An appreciable number (15) of flowering plants, specially within the Solanaceae family, have been shown to contain vitamin D 3 and its hydroxylated derivatives, including 1α,25(OH) 2 -vitamin D 3 [1α,25(OH)D 3 ], a pluripotent hormone in animals. These secosteroids have also been detected in members of the Cucurbitaceae, Fabaceae and Poaceae families. On the basis of recent cladistic analysis it is possible to predict that the synthesis of these compounds is a characteristic of the Angiosperms. Highly specific and sensitive bioassays and analytical procedures of high resolution are now available which may allow confirmation of this hypothesis. Solanum glaucophyllum is the species which accumulates 1α,25(OH)D 3 to the greatest extent. The metabolite is distributed among different tissues of the plant as a free steroid or glycoside derivatives. The presence of 7-dehydrocholesterol, vitamin D 3 , 25(OH)D 3 and 1α,25(OH) 2 D 3 in S. glaucophyllum and other species has been unequivocally demonstrated. Moreover, recent studies with radioactive precursors, protein immunoblot and RNA hybridization analysis suggest the presence of specific hydroxylases for vitamin D 3 and 25(OH)D 3 structurally related to the corresponding enzymes in vertebrates. It appears then that plants possess a similar synthetic route to 1α,25(OH) 2 D 3 as in animals. However, of unique biological importance, evidence obtained with S. glaucophyllum and Nicotiana glauca supports the operation of a non-photolytic reaction of vitamin D 3 synthesis. Both intermediates and enzymes of the vitamin D 3 pathway can be detected in cell and tissue cultures thus affording a convenient experimental model for studies on its molecular characterization and regulation. This is also endowed with biotechnological significance. Vitamin D 3 compounds may play a function in Angiosperms. Studies in vitro have shown that, like mitogenic plant hormones, they stimulate root growth and differentiation through activation of the Ca 2+ messenger system. There are vitamin D 3 and 1α,25(OH) 2 D 3 binding proteins in plants which exhibit characteristics suggesting a regulatory function. It remains to be established whether these binders act as true steroid hormone receptors.

  • Presence of estrogens and estrogen receptor-like proteins in Solanum glaucophyllum.
    Biochemical and biophysical research communications, 2001
    Co-Authors: Lorena Magdalena Milanesi, Paula V. Monje, Ricardo Boland
    Abstract:

    Abstract The synthesis of mammalian steroid hormones by plants has been reported. However, their physiological role in plants is controversial. The existence of receptor molecules as those of animal cells could provide clues into a possible steroid mechanism of action. Solanum glaucophyllum callus cultures were found to contain not only 17β-estradiol and estrone but also abundant estrogen binding sites. These sites were specific for 17β-estradiol (∼550 fmol/mg protein) and could also be competed by the known estrogen receptor (ER) agonist diethylstilbestrol. Antibodies directed against specific sequences of the classical ER α isoform, labelled a ∼67 kDa band which comigrated with the mammalian ER α antigen. ER α-like proteins were tested positive as estrogen binders in Ligand blot experiments using 17β-estradiol macromolecular derivatives as ligands. Our results provide first evidences on the existence of estrogen binding proteins structurally related to the mammalian ER α subtype in a higher plant system.

  • Effect of culture conditions on the synthesis of vitamin D3 metabolites in Solanum glaucophyllum grown in vitro
    Phytochemistry, 2001
    Co-Authors: Alejandro Carlos Curino, Lorena Magdalena Milanesi, Mario Skliar, Silvia Benassati, Ricardo Boland
    Abstract:

    Abstract In cultured Solanum glaucophyllum we have recently described the operation of a light-independent pathway of 1α,25-dihydroxy-vitamin D3 (1α,25(OH)2D3) biosynthesis which involves similar intermediates as in vertebrates. In this work we investigated factors influencing the formation of 1α,25(OH)2D3 and related sterols in S. glaucophyllum grown in vitro in darkness. Callus tissue and cells cultured in Murashige and Skoog medium in the absence of light were employed. Lipids were extracted with chloroform–methanol. The remaining water soluble fraction was incubated with β-glucosidase to release vitamin D3 compounds from their glycoconjugated derivatives followed by organic solvent extraction. Vitamin D3 derivatives were isolated by Sephadex LH-20 and high-performance liquid chromatography (HPLC). HPLC or competitive protein binding assays with intestine 1α,25(OH)2D3 receptor and serum vitamin D binding protein were used to quantify the metabolites. The levels of 1α,25(OH)2D3 in calli varied according to the tissue explant origin, e.g. stem>leaf>fruit. For all organs, the metabolite was mainly present as free sterol (>80% of total). There were larger amounts of 25(OH)D3 than 1α,25(OH)2D3. It was found that Ca2+, auxin and kinetin are important factors upregulating 1α,25(OH)2D3 synthesis in S. glaucophyllum tissue and cells. Irradiation with UV light increased vitamin D3 but not 1α,25(OH)2D3 levels. In agreement with these results, incubation of cells with [3H]25(OH)D3 revealed a low conversion rate to [3H]1α,25(OH)2D3. The operation of a light-dependent pathway formation of vitamin D3 coupled to higher expression of 25(OH)D3-1α-hydroxylase may account for the large concentrations of 1α,25(OH)2D3 normally found in differentiated field-grown plants.

Ana Russo De Boland - One of the best experts on this subject based on the ideXlab platform.

  • 1α,25(OH)2D3-glycosides from Solanum glaucophyllum leaves extract induce myoblasts differentiation through p38 MAPK and AKT activation.
    Biology open, 2018
    Co-Authors: Ana Paula Irazoqui, Pablo De Genaro, Claudia Buitrago, Heinrich Bachmann, Verónica González-pardo, Ana Russo De Boland
    Abstract:

    We have previously shown that Solanum glaucophyllum leaves extract (SGE) increases VDR protein levels and promotes myoblast differentiation. Here, we investigated whether p38 MAPK and AKT are involved in SGE actions. Cell cycle studies showed that SGE prompted a peak of S-phase followed by an arrest in the G0/G1-phase through p38 MAPK. Time course studies showed that p38 MAPK and AKT phosphorylation statically increased by SGE (10 nM) or synthetic 1α,25(OH) 2 D 3 (1 nM) treatment. Furthermore, p38 MAPK and AKT inhibitors, SB203580 and LY294002, respectively, suppressed myoblasts fusion induced by SGE or synthetic 1α,25(OH) 2 D 3 . We have also studied differentiation genes by qRT-PCR. myoD1 mRNA increased significantly by SGE (24-72 h) or 1α,25(OH) 2 D 3 (24 h) treatment. mRNA expression of myogenin also increased upon SGE or 1α,25(OH) 2 D 3 treatment. Finally, MHC2b mRNA expression, a late differentiation marker, increased significantly by both compounds at 72 h compared to control. Taken together, these results suggest that SGE, as synthetic 1α,25(OH) 2 D 3, promotes myotube formation through p38 MAPK and AKT activation.

  • 1α,25(OH)2D3-glycosides from Solanum glaucophyllum leaves extract induce myoblasts differentiation through p38 MAPK and AKT activation
    The Company of Biologists, 2018
    Co-Authors: Ana Paula Irazoqui, Pablo De Genaro, Claudia Buitrago, Heinrich Bachmann, Verónica González-pardo, Ana Russo De Boland
    Abstract:

    We have previously shown that Solanum glaucophyllum leaf extract (SGE) increases VDR protein levels and promotes myoblast differentiation. Here, we investigated whether p38 MAPK and AKT are involved in SGE actions. Cell-cycle studies showed that SGE prompted a peak of S-phase followed by an arrest in the G0/G1-phase through p38 MAPK. Time course studies showed that p38 MAPK and AKT phosphorylation were statistically increased by SGE (10 nM) or synthetic 1α,25(OH)2D3 (1 nM) treatment. Furthermore, p38 MAPK and AKT inhibitors, SB203580 and LY294002 respectively, suppressed myoblasts fusion induced by SGE or synthetic 1α,25(OH)2D3. We have also studied differentiation genes by qRT-PCR. myoD1 mRNA increased significantly by SGE (24–72 h) or 1α,25(OH)2D3 (24 h) treatment. mRNA expression of myogenin also increased upon SGE or 1α,25(OH)2D3 treatment. Finally, MHC2b mRNA expression, a late differentiation marker, was increased significantly by both compounds at 72 h compared to control. Taken together, these results suggest that SGE, as synthetic 1α,25(OH)2D3, promotes myotube formation through p38 MAPK and AKT activation

  • in vitro effects of 1α 25 oh 2d3 glycosides from solbone a Solanum glaucophyllum leaves extract herbonis ag compared to synthetic 1α 25 oh 2d3 on myogenesis
    Steroids, 2016
    Co-Authors: Valeria Gili, Ricardo Boland, Verónica González Pardo, Ana Carolina Ronda, Pablo De Genaro, Heini Bachmann, Ana Russo De Boland
    Abstract:

    The presence of glycoside derivatives of 1α,25(OH)2D3 endows plants to gradual release of the free bioactive form of 1α,25(OH)2D3 from its glycoconjugates by endogenous animal tissue glycosidases. This results in increased half-life of the hormone in blood when purified plant fractions are administered for therapeutic purposes. In this work, we evaluated the role 1α,25(OH)2D3-glycosides enriched natural product (Solbone A) from Solanum glaucophyllum leaf extract compared with synthetic 1α,25(OH)2D3 on myogenic differentiation in C2C12 myoblasts. For these, differentiation markers and myogenic parameters were studied in C2C12 myoblasts. Results showed that Solbone A, likewise the synthetic hormone, increased creatine kinase activity at day 2 after differentiation induction (60%, p<0.05). Solbone A and synthetic 1α,25(OH)2D3 increased vitamin D3 receptor protein expression at 10nM (50% and 30%, respectively) and the transcription factor myogenin (80%, p<0.05). However, tropomyosin expression was not affected by both compounds. In addition, myosin heavy chain (MHC) protein expression was increased 30% at day 2 of differentiation. Solbone A or synthetic 1α,25(OH)2D3 had no effects on myogenin nor MHC cell localization. Cellular mass increased with myogenesis progression, being Solbone A more effective than synthetic 1α,25(OH)2D3. Finally, Solbone A, as well as synthetic 1α,25(OH)2D3, augmented the index fusion of cultured muscle fibers. In conclusion, these results demonstrated that Solbone A exhibit at least equal or greater effects on early myoblast differentiation as synthetic hormone, suggesting that plant glycosides could be an effective, accessible and cheaper substitute for synthetic 1α,25(OH)2D3 to promote muscle growth.

  • In vitro effects of 1α,25(OH)2D3-glycosides from Solbone A (Solanum glaucophyllum leaves extract; Herbonis AG) compared to synthetic 1α,25(OH)2D3 on myogenesis
    Steroids, 2016
    Co-Authors: Valeria Gili, Ricardo Boland, Verónica González Pardo, Ana Carolina Ronda, Pablo De Genaro, Heini Bachmann, Ana Russo De Boland
    Abstract:

    The presence of glycoside derivatives of 1α,25(OH)2D3 endows plants to gradual release of the free bioactive form of 1α,25(OH)2D3 from its glycoconjugates by endogenous animal tissue glycosidases. This results in increased half-life of the hormone in blood when purified plant fractions are administered for therapeutic purposes. In this work, we evaluated the role 1α,25(OH)2D3-glycosides enriched natural product (Solbone A) from Solanum glaucophyllum leaf extract compared with synthetic 1α,25(OH)2D3 on myogenic differentiation in C2C12 myoblasts. For these, differentiation markers and myogenic parameters were studied in C2C12 myoblasts. Results showed that Solbone A, likewise the synthetic hormone, increased creatine kinase activity at day 2 after differentiation induction (60%, p

P. A. Fontana - One of the best experts on this subject based on the ideXlab platform.

  • Effects of Solanum glaucophyllum toxicity on cell proliferation and apoptosis in the small and large intestine of rabbits.
    Research in veterinary science, 2011
    Co-Authors: Carolina Natalia Zanuzzi, Eduardo Juan Gimeno, Enrique Leo Portiansky, P. A. Fontana, Fabián Nishida, Claudio Gustavo Barbeito
    Abstract:

    Vitamin D regulates mineral homeostases and enterocyte proliferation and differentiation. Hypervitaminosis D generates changes in cell proliferation, differentiation and apoptosis in several organs. We analysed morphometric parameters and proliferative and apoptotic indices in the intestinal epithelium of rabbits with hypervitaminosis D induced by the chronic treatment with the calcinogenic plant Solanum glaucophyllum. Rabbits were treated for 15 or 30 days. A group was treated for 15 days and led to possible recovery for 30 days. Another group was nutritionally restricted for 30 days. Morphological, morphometric, proliferative and apoptotic changes were found in the treated animals. Mild atrophy and reduced proliferation was found in the jejunum and ileum. Apoptosis increased in the crypts of the ileum and in the superficial epithelium and crypts of the rectum. Most of the alterations were partially recovered. The possible involvement in these changes of the hypervitaminosis D-like state induced by S. glaucophyllum is discussed.

  • Glycoconjugate histochemistry in the small and large intestine of normal and Solanum glaucophyllum-intoxicated rabbits
    Research in veterinary science, 2010
    Co-Authors: Carolina Natalia Zanuzzi, Enrique Leo Portiansky, P. A. Fontana, Claudio Gustavo Barbeito, M. L. Ortiz, Facundo A. Lozza, Eduardo Juan Gimeno
    Abstract:

    Vitamin D participates in mineral homeostasis, immunomodulation, cell growth and differentiation. The leaves of Solanum glaucophyllum contain high levels of 1,25-dihydroxyvitamin D3 as glycoside derivatives and their chronic ingestion generates a hypervitaminosis D-like state. We analyzed changes on carbohydrate expression as a cell differentiation indicator on samples of the small and large intestine of S. glaucophyllum-intoxicated rabbits, using conventional and lectin histochemistry. Male New Zealand white rabbits were intoxicated with S. glaucophyllum during two or four weeks and killed the day after. A group of animals ("possibly recovered group") were intoxicated during 15 days and killed at day 45 of the beginning of the experiment. We found changes in the lectin binding pattern in the small and large intestine of the intoxicated rabbits. Some of these changes were reverted in the possibly recovered group. Vitamin D could be a new regulator factor of the intestinal glycosylation process.

  • Paneth cells: histochemical and morphometric study in control and Solanum glaucophyllum intoxicated rabbits.
    European journal of histochemistry : EJH, 2009
    Co-Authors: Carolina Natalia Zanuzzi, P. A. Fontana, Enrique Leo Portiansky, Claudio Gustavo Barbeito, Eduardo Juan Gimeno
    Abstract:

    The intestinal epithelium has a critical roll in host defence. One specialised cell type involved in this function is the Paneth cell, which secretes many substances with antimicrobial properties in response to different stimuli. Under pathological conditions, changes in the Paneth cell number, morphology and location as well as in granule number, morphology and composition have been reported. In the normal animal, 1,25-dihydroxyvitamin D3 participates in the maintenance of mineral homeostasis, immunomodulation and cell proliferation and differentiation. Solanum glaucophyllum, a calcinogenic plant containing high levels of 1,25-dihydroxyvitamin D3, is responsible for a condition known as enzootic calcinosis in ruminants, characterised by loss of body condition and mineralization of soft tissues. Using and established rabbit model, this study analyses the changes that rabbit Paneth cells undergo during intoxication with S. glaucophyllum. Male New Zealand white rabbits were experimentally intoxicated with S. glaucophyllum for 15 or 30 days. Lectin, immunohistochemical and morphometric studies were carried out on Paneth cells from samples of jejunum. SBA, DBA and WGA lectins bound to Paneth cells-granules in both normal and intoxicated rabbits, with more heterogenity in the labelling of granules from intoxicated rabbits. Paneth cells in both groups were immunonegative for lysosyme. A time and dose-dependent increase in the size and number of Paneth cells was found in both intoxicated groups. We suggest that the changes described in these cells may be directly or indirectly induced by S. glaucophyllum intoxication.

  • Thymic atrophy in cattle poisoned with Solanum glaucophyllum
    Pesquisa Veterinária Brasileira, 2009
    Co-Authors: P. A. Fontana, Eduardo Juan Gimeno, Carolina Natalia Zanuzzi, Claudio Gustavo Barbeito, Enrique Leo Portiansky
    Abstract:

    Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing "Enzootic Calcinosis" in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.

  • Thymic atrophy in cattle poisoned with Solanum glaucophyllum Atrofia do timo em bovinos intoxicados por Solanum glaucophyllum
    Colégio Brasileiro de Patologia Animal (CBPA), 2009
    Co-Authors: P. A. Fontana, Carolina Natalia Zanuzzi, Claudio Gustavo Barbeito, Eduardo J. Gimeno, Enrique L. Portiansky
    Abstract:

    Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing "Enzootic Calcinosis" in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.Solanum glaucophyllum (Sg) [= S. malacoxylon] é uma planta calcinogênica que induz "Calcinose Enzoótica" em bovinos. O 1,25-dihidroxivitamina D3, seu principal agente tóxico, regula o metabolismo ósseo, o metabolismo de cálcio e também mostra efeitos na imunomodulação. Precursores de timócitos derivados da medula óssea se diferenciam em linfócitos T maduros. A diferenciação da maioria dos linfócitos T é caracterizada pela expressão variável de moléculas de receptores CD4/CD8 e densidade aumentada dos receptores antigênicos de superfície de células T. Alem disso, há mudanças no padrão de glicosilação de glicolipídeos na superfície celular ou de glicoproteínas. Timócitos mostram uma influência de retro alimentação em células tímicas não-linfóides. Foram analisadas modificações induzidas pelo Sg em linfócitos T e células tímicas não-linfóides de bovinos. Novilhas foram divididas em 5 grupos (controle, intoxicadas com Sg durante 15, 30 ou 60 dias, e grupo provavelmente recuperado). As diferentes populações celulares das novilhas experimentais foram caracterizadas com técnicas histoquímicas, imuno-histoquímicas, lectina-histoquímicas e morfométricas. As novilhas intoxicadas com Sg mostraram uma atrofia cortical progressiva que foi caracterizada usando a lectina aglutinina de amendoim (PNA) que reconhece timócitos imaturos. Estes animais também aumentaram as células não-linfóides tímicas por unidade de área, detectadas com a técnica de Picrosirius, lectinas WGA e DBA e anticorpos antipancitoqueratina e anti-S-100. A atrofia de timo observada nos animais intoxicados foi semelhante àquela do processo de envelhecimento fisiológico. Após supressão da intoxicação, foi observado um efeito de reversão nestas mudanças. Estes resultados sugerem que a intoxicação por Sg induza a alteração observada no timo diretamente, pela ação de 1,25-dihidroxivitamina D3, ou indiretamente, pela ação da hipercalcemia

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  • Effects of Solanum glaucophyllum toxicity on cell proliferation and apoptosis in the small and large intestine of rabbits.
    Research in veterinary science, 2011
    Co-Authors: Carolina Natalia Zanuzzi, Eduardo Juan Gimeno, Enrique Leo Portiansky, P. A. Fontana, Fabián Nishida, Claudio Gustavo Barbeito
    Abstract:

    Vitamin D regulates mineral homeostases and enterocyte proliferation and differentiation. Hypervitaminosis D generates changes in cell proliferation, differentiation and apoptosis in several organs. We analysed morphometric parameters and proliferative and apoptotic indices in the intestinal epithelium of rabbits with hypervitaminosis D induced by the chronic treatment with the calcinogenic plant Solanum glaucophyllum. Rabbits were treated for 15 or 30 days. A group was treated for 15 days and led to possible recovery for 30 days. Another group was nutritionally restricted for 30 days. Morphological, morphometric, proliferative and apoptotic changes were found in the treated animals. Mild atrophy and reduced proliferation was found in the jejunum and ileum. Apoptosis increased in the crypts of the ileum and in the superficial epithelium and crypts of the rectum. Most of the alterations were partially recovered. The possible involvement in these changes of the hypervitaminosis D-like state induced by S. glaucophyllum is discussed.

  • Glycoconjugate histochemistry in the small and large intestine of normal and Solanum glaucophyllum-intoxicated rabbits
    Research in veterinary science, 2010
    Co-Authors: Carolina Natalia Zanuzzi, Enrique Leo Portiansky, P. A. Fontana, Claudio Gustavo Barbeito, M. L. Ortiz, Facundo A. Lozza, Eduardo Juan Gimeno
    Abstract:

    Vitamin D participates in mineral homeostasis, immunomodulation, cell growth and differentiation. The leaves of Solanum glaucophyllum contain high levels of 1,25-dihydroxyvitamin D3 as glycoside derivatives and their chronic ingestion generates a hypervitaminosis D-like state. We analyzed changes on carbohydrate expression as a cell differentiation indicator on samples of the small and large intestine of S. glaucophyllum-intoxicated rabbits, using conventional and lectin histochemistry. Male New Zealand white rabbits were intoxicated with S. glaucophyllum during two or four weeks and killed the day after. A group of animals ("possibly recovered group") were intoxicated during 15 days and killed at day 45 of the beginning of the experiment. We found changes in the lectin binding pattern in the small and large intestine of the intoxicated rabbits. Some of these changes were reverted in the possibly recovered group. Vitamin D could be a new regulator factor of the intestinal glycosylation process.

  • Paneth cells: histochemical and morphometric study in control and Solanum glaucophyllum intoxicated rabbits.
    European journal of histochemistry : EJH, 2009
    Co-Authors: Carolina Natalia Zanuzzi, P. A. Fontana, Enrique Leo Portiansky, Claudio Gustavo Barbeito, Eduardo Juan Gimeno
    Abstract:

    The intestinal epithelium has a critical roll in host defence. One specialised cell type involved in this function is the Paneth cell, which secretes many substances with antimicrobial properties in response to different stimuli. Under pathological conditions, changes in the Paneth cell number, morphology and location as well as in granule number, morphology and composition have been reported. In the normal animal, 1,25-dihydroxyvitamin D3 participates in the maintenance of mineral homeostasis, immunomodulation and cell proliferation and differentiation. Solanum glaucophyllum, a calcinogenic plant containing high levels of 1,25-dihydroxyvitamin D3, is responsible for a condition known as enzootic calcinosis in ruminants, characterised by loss of body condition and mineralization of soft tissues. Using and established rabbit model, this study analyses the changes that rabbit Paneth cells undergo during intoxication with S. glaucophyllum. Male New Zealand white rabbits were experimentally intoxicated with S. glaucophyllum for 15 or 30 days. Lectin, immunohistochemical and morphometric studies were carried out on Paneth cells from samples of jejunum. SBA, DBA and WGA lectins bound to Paneth cells-granules in both normal and intoxicated rabbits, with more heterogenity in the labelling of granules from intoxicated rabbits. Paneth cells in both groups were immunonegative for lysosyme. A time and dose-dependent increase in the size and number of Paneth cells was found in both intoxicated groups. We suggest that the changes described in these cells may be directly or indirectly induced by S. glaucophyllum intoxication.

  • Thymic atrophy in cattle poisoned with Solanum glaucophyllum
    Pesquisa Veterinária Brasileira, 2009
    Co-Authors: P. A. Fontana, Eduardo Juan Gimeno, Carolina Natalia Zanuzzi, Claudio Gustavo Barbeito, Enrique Leo Portiansky
    Abstract:

    Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing "Enzootic Calcinosis" in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.

  • Thymic atrophy in cattle poisoned with Solanum glaucophyllum Atrofia do timo em bovinos intoxicados por Solanum glaucophyllum
    Colégio Brasileiro de Patologia Animal (CBPA), 2009
    Co-Authors: P. A. Fontana, Carolina Natalia Zanuzzi, Claudio Gustavo Barbeito, Eduardo J. Gimeno, Enrique L. Portiansky
    Abstract:

    Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing "Enzootic Calcinosis" in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.Solanum glaucophyllum (Sg) [= S. malacoxylon] é uma planta calcinogênica que induz "Calcinose Enzoótica" em bovinos. O 1,25-dihidroxivitamina D3, seu principal agente tóxico, regula o metabolismo ósseo, o metabolismo de cálcio e também mostra efeitos na imunomodulação. Precursores de timócitos derivados da medula óssea se diferenciam em linfócitos T maduros. A diferenciação da maioria dos linfócitos T é caracterizada pela expressão variável de moléculas de receptores CD4/CD8 e densidade aumentada dos receptores antigênicos de superfície de células T. Alem disso, há mudanças no padrão de glicosilação de glicolipídeos na superfície celular ou de glicoproteínas. Timócitos mostram uma influência de retro alimentação em células tímicas não-linfóides. Foram analisadas modificações induzidas pelo Sg em linfócitos T e células tímicas não-linfóides de bovinos. Novilhas foram divididas em 5 grupos (controle, intoxicadas com Sg durante 15, 30 ou 60 dias, e grupo provavelmente recuperado). As diferentes populações celulares das novilhas experimentais foram caracterizadas com técnicas histoquímicas, imuno-histoquímicas, lectina-histoquímicas e morfométricas. As novilhas intoxicadas com Sg mostraram uma atrofia cortical progressiva que foi caracterizada usando a lectina aglutinina de amendoim (PNA) que reconhece timócitos imaturos. Estes animais também aumentaram as células não-linfóides tímicas por unidade de área, detectadas com a técnica de Picrosirius, lectinas WGA e DBA e anticorpos antipancitoqueratina e anti-S-100. A atrofia de timo observada nos animais intoxicados foi semelhante àquela do processo de envelhecimento fisiológico. Após supressão da intoxicação, foi observado um efeito de reversão nestas mudanças. Estes resultados sugerem que a intoxicação por Sg induza a alteração observada no timo diretamente, pela ação de 1,25-dihidroxivitamina D3, ou indiretamente, pela ação da hipercalcemia