Pub Date : 2008-08-01DOI: 10.32604/BIOCELL.2008.32.175
N. Ocarino, A. Bozzi, R. Pereira, N. Breyner, V. L. Silva, P. Castanheira, A. Goes, R. Serakides
4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) is a DNA dye widely used to mark and trace stem cells in therapy. We here studied the effect of DAPI staining on the behavior of mesenchymal stem cells cultured in either a control, non-osteogenic medium or in an osteogenic differentiation medium. In the control medium, the number of stem cells/field, as well as the number of fluorescent cells/field increased up to the sixth day in both control and DAPI-treated cultures. Afterwards, both the number of fluorescent cells and their fluorescence intensity decreased. Control cells were fusiform and with some long extensions that apparently linked them to neighboring cells, while DAPI-treated cells were mostly round cells with fine and short extensions. The trypan-blue exclusion method showed 99% cell viability in both groups, however, both alkaline phosphatase activity and the thiazolyl blue formazan assay (indicative of mitochondrial metabolism) gave significantly lower values in DAPI-marked cells. The mitochondrial mass, as indicated by specific staining and flow cytometry, showed no differences between groups. Mesenchymal stem cells gave origin to mineralized nodules in the osteogenic differentiation medium and there were not DAPI-marked cells on the ninth day of culture. Alkaline phosphatase activity, viability assay and number of cells/field and of mineralized nodules/field were similar in both groups. So, DAPI treatment did not change cell viability and proliferation during osteogenic differentiation of mesenchymal stem cells. However, since these cells loose DAPI marking after 9 days in osteogenic cultures suggests that DAPI may not be an effective marker for mesenchymal stem cells implanted in bone tissue for long periods.
{"title":"Behavior of mesenchymal stem cells stained with 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) in osteogenic and non osteogenic cultures.","authors":"N. Ocarino, A. Bozzi, R. Pereira, N. Breyner, V. L. Silva, P. Castanheira, A. Goes, R. Serakides","doi":"10.32604/BIOCELL.2008.32.175","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.175","url":null,"abstract":"4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) is a DNA dye widely used to mark and trace stem cells in therapy. We here studied the effect of DAPI staining on the behavior of mesenchymal stem cells cultured in either a control, non-osteogenic medium or in an osteogenic differentiation medium. In the control medium, the number of stem cells/field, as well as the number of fluorescent cells/field increased up to the sixth day in both control and DAPI-treated cultures. Afterwards, both the number of fluorescent cells and their fluorescence intensity decreased. Control cells were fusiform and with some long extensions that apparently linked them to neighboring cells, while DAPI-treated cells were mostly round cells with fine and short extensions. The trypan-blue exclusion method showed 99% cell viability in both groups, however, both alkaline phosphatase activity and the thiazolyl blue formazan assay (indicative of mitochondrial metabolism) gave significantly lower values in DAPI-marked cells. The mitochondrial mass, as indicated by specific staining and flow cytometry, showed no differences between groups. Mesenchymal stem cells gave origin to mineralized nodules in the osteogenic differentiation medium and there were not DAPI-marked cells on the ninth day of culture. Alkaline phosphatase activity, viability assay and number of cells/field and of mineralized nodules/field were similar in both groups. So, DAPI treatment did not change cell viability and proliferation during osteogenic differentiation of mesenchymal stem cells. However, since these cells loose DAPI marking after 9 days in osteogenic cultures suggests that DAPI may not be an effective marker for mesenchymal stem cells implanted in bone tissue for long periods.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127596203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-08-01DOI: 10.32604/BIOCELL.2008.32.185
A. Ferreira, M. Mehanna, C. Prado
The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.
{"title":"Morphologic and morphometric analysis of testis of Pseudis limellum (Cope, 1862) (Anura, Hylidae) during the reproductive cycle in the Pantanal, Brazil.","authors":"A. Ferreira, M. Mehanna, C. Prado","doi":"10.32604/BIOCELL.2008.32.185","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.185","url":null,"abstract":"The spermatogenesis of Pseudis limellum, from the Southern Pantanal, Brazil, was studied from July 1995 to May 1996, through histological sections of the testis. The cells could be differentiated as: primary spermatogonia, large cells, generally with bilobed nucleus; secondary spermatogonia, smaller cells, with darker cytoplasm, chromatin of radial form; primary and secondary spermatocytes, differentiated according to the different stages of the nucleus during the successive cells divisions. Furthermore, we observed cells in process of morphologic differentiation: rounded spermatids much smaller, with nucleus containing chromatin in compacting process and cytoplasm reduction; elongated spermatids, generally parallel organized in well defined bundles, with the anterior region directed toward the periphery of the seminiferous tubule and the tail directed toward the lumen. Spermatozoa are free in the lumen of the seminiferous tubule. All the cells are organized as cysts, which are supported by a large amount of Sertoli cells. The spermatogenesis in P. limellum is very similar to that of other anurans, but peculiarities were observed regarding the organization of the germ cells, the great amount of free Sertoli cells in the lumen of testis collected in May, and the long cytoplasmatic extensions of the cells bearing pigments and involving the seminiferous tubule. The diameter of the seminiferous tubule (SD) exhibited an annual mean of 251.79 +/- 37.57 microm. Spermatozoa number by seminiferous tubule (SN) exhibited an annual mean of 306.66 +/- 39.83, also with higher and lower values at each month. Variations in SD and SN were not significantly correlated with climatic variables. In this species, reproduction occurs throughout the year in ponds and flooded areas, despite the seasonal climate of the Pantanal. Although males varied in their annual reproductive activity, they were considered potentially reproductive in all months throughout the year.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"15 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132365963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-08-01DOI: 10.32604/BIOCELL.2008.32.169
I. Novak, H. Cabral
We had previously found in autologous human leukocyte cultures, in which dead neutrophils phagocytosis by macrophages occur, macrophages and T CD4 lymphocytes perform a selective cell-cell interaction showing many figures of either one, two or several T- lymphocytes adhering to a central macrophage were seen. Considering that antigen presentation would be necessary for the formation of these immune synapses, we attempted to block rosette formation (i.e., the formation of macrophage associations with at least three lymphocytes) by interfering with both antigen processing and presentation. Culture samples of autologous leukocytes from 7 healthy donors were subjected to either brefeldin A, chloroquine or to an anti-HLA DR antibody. Rosette formation was significantly inhibited in the treated samples (either with brefeldin A, chloroquine or the anti- HLA DR; ANOVA, p<0.001, as compared with the untreated controls). It is concluded that interference with antigen processing and presentation precludes the formation of these cell-cell interactions.
{"title":"Rosette formation by macrophages with adhered T lymphocytes is precluded by inhibitors of antigen processing and presentation.","authors":"I. Novak, H. Cabral","doi":"10.32604/BIOCELL.2008.32.169","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.169","url":null,"abstract":"We had previously found in autologous human leukocyte cultures, in which dead neutrophils phagocytosis by macrophages occur, macrophages and T CD4 lymphocytes perform a selective cell-cell interaction showing many figures of either one, two or several T- lymphocytes adhering to a central macrophage were seen. Considering that antigen presentation would be necessary for the formation of these immune synapses, we attempted to block rosette formation (i.e., the formation of macrophage associations with at least three lymphocytes) by interfering with both antigen processing and presentation. Culture samples of autologous leukocytes from 7 healthy donors were subjected to either brefeldin A, chloroquine or to an anti-HLA DR antibody. Rosette formation was significantly inhibited in the treated samples (either with brefeldin A, chloroquine or the anti- HLA DR; ANOVA, p<0.001, as compared with the untreated controls). It is concluded that interference with antigen processing and presentation precludes the formation of these cell-cell interactions.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123502645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-08-01DOI: 10.32604/BIOCELL.2008.32.207
Patrícia Carvalho de Souza, A. Khayat, I. C. Seligmann, R. R. Rodriguez Burbano
The collared peccary (Tayassu tajacu) is widely distributed over the American continent, being found from the south of the USA to the north of Argentina. In Brazil, it is spread all over the country, being one of the potential species to be raised in captivity. Therefore, the cytogenetic techniques could be a potential tool for reproductive monitoring of animals raised in captivity, mainly when destined for commercial purposes. This study had the objective of determining the chromosome number of two populations raised in captivity and characterizing them by GTG banding. For this purpose, an analysis was made of mitotic metaphases obtained from lymphocyte cultures made from blood samples of 11 animals, six of which from the Northeast and five from the North of Brazil. The results of this analysis showed the same karyotype pattern for the species (2n=30 chromosomes and NF=48), besides corresponding to the South American pattern of the species, i.e., without a translocation between autosomes 1 and 8, chromosome X acrocentric, and no differences were found between the two populations studied. However, chromosomal polymorphisms were observed compared to data from the literature on populations from North and South America.
{"title":"Chromosome comparison between populations of the collared peccary, Tayassu tajacu, raised in captivity.","authors":"Patrícia Carvalho de Souza, A. Khayat, I. C. Seligmann, R. R. Rodriguez Burbano","doi":"10.32604/BIOCELL.2008.32.207","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.207","url":null,"abstract":"The collared peccary (Tayassu tajacu) is widely distributed over the American continent, being found from the south of the USA to the north of Argentina. In Brazil, it is spread all over the country, being one of the potential species to be raised in captivity. Therefore, the cytogenetic techniques could be a potential tool for reproductive monitoring of animals raised in captivity, mainly when destined for commercial purposes. This study had the objective of determining the chromosome number of two populations raised in captivity and characterizing them by GTG banding. For this purpose, an analysis was made of mitotic metaphases obtained from lymphocyte cultures made from blood samples of 11 animals, six of which from the Northeast and five from the North of Brazil. The results of this analysis showed the same karyotype pattern for the species (2n=30 chromosomes and NF=48), besides corresponding to the South American pattern of the species, i.e., without a translocation between autosomes 1 and 8, chromosome X acrocentric, and no differences were found between the two populations studied. However, chromosomal polymorphisms were observed compared to data from the literature on populations from North and South America.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"72 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133914379","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-08-01DOI: 10.32604/BIOCELL.2008.32.195
Raquel Alves dos Santos, T. Cabral, I. Cabral, L. M. Antunes, Cristiane Pontes Andrade, Plínio Cerqueira dos Santos Cardoso, Marcelo de Oliveira Bahia, C. Pessoa, J. L. Martins do Nascimento, R. R. Rodriguez Burbano, C. Takahashi
Physalis angulata L (Solanaceae) is a medicinal plant from North of Brazil, whose different extracts and infusions are commonly used in the popular medicine for the treatment of malaria, asthma, hepatitis, dermatitis and rheumatism. However, the genotoxic effects of P. angulata on human cells is not well known. The main purpose of the present study was to evaluate the in vitro genotoxic effects of aqueous extract of P. angulata using the comet assay and the micronucleus assay in human lymphocytes provided from 6 healthy donors. Treatments with P. angulata extracts were performed in vitro in order to access the extent of DNA damage. The comet assay has shown that treatments with P. angulata at 0.5, 1.0, 2.0, 3.0 and 6.0 microg/mL in culture medium were genotoxic. Lymphocytes treated with P. angulata at the concentrations of 3.0 and 6.0 microg/mL in culture medium showed a statistically significant increase in the frequency of micronucleus (p<0.05), however, the cytokinesis blocked proliferation index (CBPI) was not decreased after P. angulata treatment. In conclusion, the present work demonstrated the genotoxic effects of P. angulata extract on human lymphocytes in vitro.
{"title":"Genotoxic effect of Physalis angulata L. (Solanaceae) extract on human lymphocytes treated in vitro.","authors":"Raquel Alves dos Santos, T. Cabral, I. Cabral, L. M. Antunes, Cristiane Pontes Andrade, Plínio Cerqueira dos Santos Cardoso, Marcelo de Oliveira Bahia, C. Pessoa, J. L. Martins do Nascimento, R. R. Rodriguez Burbano, C. Takahashi","doi":"10.32604/BIOCELL.2008.32.195","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.195","url":null,"abstract":"Physalis angulata L (Solanaceae) is a medicinal plant from North of Brazil, whose different extracts and infusions are commonly used in the popular medicine for the treatment of malaria, asthma, hepatitis, dermatitis and rheumatism. However, the genotoxic effects of P. angulata on human cells is not well known. The main purpose of the present study was to evaluate the in vitro genotoxic effects of aqueous extract of P. angulata using the comet assay and the micronucleus assay in human lymphocytes provided from 6 healthy donors. Treatments with P. angulata extracts were performed in vitro in order to access the extent of DNA damage. The comet assay has shown that treatments with P. angulata at 0.5, 1.0, 2.0, 3.0 and 6.0 microg/mL in culture medium were genotoxic. Lymphocytes treated with P. angulata at the concentrations of 3.0 and 6.0 microg/mL in culture medium showed a statistically significant increase in the frequency of micronucleus (p<0.05), however, the cytokinesis blocked proliferation index (CBPI) was not decreased after P. angulata treatment. In conclusion, the present work demonstrated the genotoxic effects of P. angulata extract on human lymphocytes in vitro.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"338 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134260635","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-08-01DOI: 10.32604/biocell.2008.32.163
J. Zanoni, Renata Virginia Fernandes Pereira
The objective of this work was to evaluate the effect of the ascorbic acid supplementation on the cellular proliferation on the ileum mucosa of diabetic rats. Fifteen 90-days rats were divided in the groups: control, diabetic and diabetic supplemented with ascorbic acid (DA). Two hours prior the sacrifice, they were injected with Vincristin. Semi-seriate histological cuts stained with HE were accomplished. About 2500 crypt cells from the intestinal mucosa were counted in order to obtain the metaphasic indexes. The height and depth of 30 villi and 30 crypts were measured for each animal, respectively. The metaphasic indexes showed no significant changes when we compared the three groups: 20.2 +/- 0.7 (control), 18 +/- 1.9 (diabetic) and 17 +/- 1.4 (DA) (p > 0.05). The values obtained from the crypts measurement were 221.2 +/- 8.5 (control), 225.3 +/- 9.5 (diabetic) and 222 +/- 34 (DA). The villi of the control, diabetic and DA animals presented the following results: 301.7 +/- 25.33, 304.8 +/- 25.63 and 322.1 +/- 45.77 respectively. The morphometric data were not different statistically (p > 0.05). Summing up, the present work showed that there was no alteration in the cellular proliferation of the ileum of diabetic-induced rats supplemented with ascorbic acid.
{"title":"Cell proliferation of the ileum intestinal mucosa of diabetic rats treated with ascorbic acid.","authors":"J. Zanoni, Renata Virginia Fernandes Pereira","doi":"10.32604/biocell.2008.32.163","DOIUrl":"https://doi.org/10.32604/biocell.2008.32.163","url":null,"abstract":"The objective of this work was to evaluate the effect of the ascorbic acid supplementation on the cellular proliferation on the ileum mucosa of diabetic rats. Fifteen 90-days rats were divided in the groups: control, diabetic and diabetic supplemented with ascorbic acid (DA). Two hours prior the sacrifice, they were injected with Vincristin. Semi-seriate histological cuts stained with HE were accomplished. About 2500 crypt cells from the intestinal mucosa were counted in order to obtain the metaphasic indexes. The height and depth of 30 villi and 30 crypts were measured for each animal, respectively. The metaphasic indexes showed no significant changes when we compared the three groups: 20.2 +/- 0.7 (control), 18 +/- 1.9 (diabetic) and 17 +/- 1.4 (DA) (p > 0.05). The values obtained from the crypts measurement were 221.2 +/- 8.5 (control), 225.3 +/- 9.5 (diabetic) and 222 +/- 34 (DA). The villi of the control, diabetic and DA animals presented the following results: 301.7 +/- 25.33, 304.8 +/- 25.63 and 322.1 +/- 45.77 respectively. The morphometric data were not different statistically (p > 0.05). Summing up, the present work showed that there was no alteration in the cellular proliferation of the ileum of diabetic-induced rats supplemented with ascorbic acid.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"109 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117192882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-04-01DOI: 10.32604/BIOCELL.2008.32.041
S. Rosenfeldt, B. Galati
Eleven Oxalis L. species from the province of Buenos Aires (Argentina) were investigated with scanning and transmission electron microscopes. We identified four different types and two subtypes of orbicules. We conclude that the close morphological similarity between these species is also reflected in their orbicules, and we suggest that the orbicules morphology may be a useful character in systematic studies.
{"title":"Orbicules diversity in Oxalis species from the province of Buenos Aires (Argentina).","authors":"S. Rosenfeldt, B. Galati","doi":"10.32604/BIOCELL.2008.32.041","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.041","url":null,"abstract":"Eleven Oxalis L. species from the province of Buenos Aires (Argentina) were investigated with scanning and transmission electron microscopes. We identified four different types and two subtypes of orbicules. We conclude that the close morphological similarity between these species is also reflected in their orbicules, and we suggest that the orbicules morphology may be a useful character in systematic studies.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"48 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"125033304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-04-01DOI: 10.32604/BIOCELL.2008.32.001
F. Capani, E. Saraceno, V. Boti, Laura Aon-Bertolino, J. C. Fernández, Fernándo Gato, M. Kruse, L. Giraldez, Mark Ellisman, H. Coirini
Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions.
{"title":"A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.","authors":"F. Capani, E. Saraceno, V. Boti, Laura Aon-Bertolino, J. C. Fernández, Fernándo Gato, M. Kruse, L. Giraldez, Mark Ellisman, H. Coirini","doi":"10.32604/BIOCELL.2008.32.001","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.001","url":null,"abstract":"Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"5 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124493381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-04-01DOI: 10.32604/BIOCELL.2008.32.027
A. Weyers, L. Ugnia, H. Ovando, N. Gorla
In the present study, the antioxidant capacity of vitamin C was examined in the liver and the kidney tissues of mice with or without ciprofloxacin (CFX) treatment. The antioxidant capacity of the vitamin was evaluated in terms of lipid hydroperoxides (LOOH) and thiobarbituric acid reactive substances (TBARs). The experimental design was 15 days of water (control and CFX groups) or vitamin C (vitamin C and vitamin C plus CFX groups) in drinking water. One dose of CFX was injected, 15 minutes before sacrifice, in the corresponding mice. The initial nmol of lipid hydroperoxides/g of tissue were 137 +/- 11 in the kidney and 145 +/- 15 in the liver, and the nmol of TBARs were 13 +/- 0.7 and 12 +/- 0.6, respectively. Pre-treatment with vitamin C reduced the levels of LOOH in the liver to 45 +/- 11 (p < 0.01) and vitamin C with CFX injection to 54 +/- 9 (p < 0.01). Vitamin C treatment also reduced the LOOH levels in the kidney roughly duplicated by CFX. Through the TBARs method we have not observed these effects. Quantification of LOOH is more sensitive than that of TBARs for estimating lipid peroxidation. CFX is used especially for urinary infections and can produce oxidative stress in the kidney. Pre-treatment with vitamin C may ameliorate this stress and also may improve the oxidative balance in the liver.
{"title":"Antioxidant capacity of vitamin C in mouse liver and kidney tissues.","authors":"A. Weyers, L. Ugnia, H. Ovando, N. Gorla","doi":"10.32604/BIOCELL.2008.32.027","DOIUrl":"https://doi.org/10.32604/BIOCELL.2008.32.027","url":null,"abstract":"In the present study, the antioxidant capacity of vitamin C was examined in the liver and the kidney tissues of mice with or without ciprofloxacin (CFX) treatment. The antioxidant capacity of the vitamin was evaluated in terms of lipid hydroperoxides (LOOH) and thiobarbituric acid reactive substances (TBARs). The experimental design was 15 days of water (control and CFX groups) or vitamin C (vitamin C and vitamin C plus CFX groups) in drinking water. One dose of CFX was injected, 15 minutes before sacrifice, in the corresponding mice. The initial nmol of lipid hydroperoxides/g of tissue were 137 +/- 11 in the kidney and 145 +/- 15 in the liver, and the nmol of TBARs were 13 +/- 0.7 and 12 +/- 0.6, respectively. Pre-treatment with vitamin C reduced the levels of LOOH in the liver to 45 +/- 11 (p < 0.01) and vitamin C with CFX injection to 54 +/- 9 (p < 0.01). Vitamin C treatment also reduced the LOOH levels in the kidney roughly duplicated by CFX. Through the TBARs method we have not observed these effects. Quantification of LOOH is more sensitive than that of TBARs for estimating lipid peroxidation. CFX is used especially for urinary infections and can produce oxidative stress in the kidney. Pre-treatment with vitamin C may ameliorate this stress and also may improve the oxidative balance in the liver.","PeriodicalId":342778,"journal":{"name":"Biocell : official journal of the Sociedades Latinoamericanas de Microscopia Electronica ... et. al","volume":"66 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134243695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2008-04-01DOI: 10.32604/biocell.2008.32.033
L. Mroginski, P. Sansberro, A. Scocchi, C. Luna, H. Rey
Tropical Ilex species have recalcitrant seeds. This work describes experiments demonstrating the feasibility of long-term conservation of Ilex brasiliensis, I. brevicuspis, I. dumosa, I. intergerrima, I. paraguariensis, I. pseudoboxus, I. taubertiana, and I. theezans through cryopreservation of zygotic rudimentary embryos at the heart developmental stage. The embryos were aseptically removed from the seeds and precultured (7 days) in the dark, at 27 +/- 2 degrees C on solidified (0.8% agar) 1/4MS medium, [consisting of quarter-strength salts and vitamins of Murashige and Skoog (1962) medium] with 3% sucrose and 0.1 mg/l Zeatin. The embryos were then encapsulated in 3% calcium alginate beads and pretreated at 24 h intervals in liquid medium supplemented with progressively increasing sucrose concentrations (0.5, 0.75 and 1 M). Beads were dehydrated for 5 h with silicagel to 25% water content (fresh weight basis) and then placed in sterile 5 ml cryovials. Then the beads were either plunged rapidly in liquid nitrogen were they were kept for 1 h (rapid cooling) or cooled at 1 degrees C min(-1) to -30 degrees C. Then the beads were immersed in liquid nitrogen for 1 h (slow cooling). The beads were rewarmed by immersion of the cryovials for 1 min in a water bath thermostated at 30 degrees C. Finally, beads were transferred onto culture medium (1/4MS, 3% sucrose, 0.1 mg/l zeatin, solidified with 0.8% agar) and incubated in a growth room at 27 +/- 2 degrees C under a 14 h light (116 micromol. m(-2) x s(-1))/ 10 h dark photoperiod. Maximum recovery percentages between 15 and 83% (depending on de the species and the treatment) were obtained with the cryopreserved embryos.
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