46 results on '"somatic tissue"'
Search Results
2. Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)
- Author
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Sakil Ahmed, Aamir Sohail, Sabiha Khatoon, Shabnam Khan, and Mohammad Khalid Saifullah
- Subjects
Bubalus bubalis ,Gastrothylax crumenifer ,glutathione S-transferase ,purification ,somatic tissue ,Animal culture ,SF1-1100 ,Veterinary medicine ,SF600-1100 - Abstract
Aim: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis). Materials and Methods: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture. Results: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5. Conclusion: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C.
- Published
- 2017
- Full Text
- View/download PDF
3. Influence of cryopreservation techniques and low concentrations of permeating cryoprotectants on the conservation of ear cartilage and skin derived from six-banded armadillos (Euphractus sexcinctus Linnaeus, 1758).
- Author
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Fernandes, Denilsa Pires, Praxedes, Érika Almeida, Viana, João Vitor da Silva, de Aquino, Leonardo Vitorino Costa, Rodrigues, Luanna Lorenna Vieira, Moura, Yasmin Beatriz França, de Oliveira, Moacir Franco, Freitas, Carlos Iberê Alves, and Pereira, Alexsandra Fernandes
- Subjects
- *
ARMADILLOS , *CRYOPROTECTIVE agents , *EAR , *CARTILAGE , *FROZEN semen , *SOMATIC cells , *ETHYLENE glycol - Abstract
Considering the importance of efficiently obtaining somatic resource banks as an ex-situ conservation strategy for wild mammals, we evaluated two techniques (slow freezing - SF and solid-surface vitrification - SSV) for the cryopreservation of ear cartilage and skin from six-banded armadillos. Additionally, we analyzed the effects of two combinations of intracellular cryoprotectants (3.0 M or 6.0 M ethylene glycol - EG and dimethyl sulfoxide - Me 2 SO) on SSV. Tissues not subjected to cryopreservation were used as controls. All samples were evaluated for morphological analysis and cell ability during culture. The thickness of the basal layer was similar to the control only for tissues derived from SF, while SSV ensured the preservation of the cartilage thickness. Moreover, fragments derived from SF and SSV, especially in the 3.0 M EG-Me 2 SO group, resulted in dermis thickness and total skin similar to the control. All cryopreserved techniques maintained normal patterns of the fibroblasts, epidermal cells, and melanocytes. While only SF-derived fragments maintained the number of degenerated chondrocytes similar to the control, no difference was observed between groups for normal chondrocytes and lacunae. Moreover, SSV maintained the collagen fibers percentage of the tissues even after warming. After culture, SF and SSV techniques were efficient for the recovery of the somatic cells in all parameters evaluated, except the day of subconfluence, which was greater for the SSV group with 6.0 M EG-Me 2 SO. In summary, SSV, especially with 3 M EG-Me 2 SO, was as efficient as SF in preserving ear skin and cartilage derived from six-banded armadillos. • Solid-surface vitrification with 3 M EG-Me 2 SO was as efficient as slow freezing in preserving skin of six-banded armadillos. • Solid-surface vitrification is suggested as the most convenient technique than slow freezing for six-banded armadillos • The efficiency of the vitrification depends on the use of lower concentrations of permeating cryoprotectants at 3 M • Our data provide an important basis for the formation of tissue bank for six-banded armadillos. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
4. Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536
- Author
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Angela Ricci, Luca Capriotti, Bruno Mezzetti, Oriano Navacchi, and Silvia Sabbadini
- Subjects
Prunus persica ,in vitro organogenesis ,somatic tissue ,STS ,carbenicillin ,cefotaxime ,Botany ,QK1-989 - Abstract
In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.
- Published
- 2020
- Full Text
- View/download PDF
5. Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758).
- Author
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Borges, Alana A., Queiroz Neta, Luiza B., Santos, Maria V. O., Oliveira, Moacir F., Silva, Alexandre R., and Pereira, Alexsandra F.
- Abstract
Copyright of Pesquisa Veterinaria Brasileira is the property of Colegio Brasileiro de Patologia Animal - CBPA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
- Published
- 2018
- Full Text
- View/download PDF
6. Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea).
- Author
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Ahmed, Sakil, Sohail, Aamir, Khatoon, Sabiha, Khan, Shabnam, and Saifullah, Mohammad Khalid
- Subjects
- *
GLUTATHIONE transferase , *SOMATIC cells , *TREMATODA , *WATER buffalo , *PRECIPITATION (Chemistry) , *GEL permeation chromatography - Abstract
Aim: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis). Materials and Methods: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture. Results: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5. Conclusion: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
7. Evolutionary Theories of Imprinting— Enough Already!
- Author
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Moore, Tom, Mills, Walter, Back, Nathan, editor, Cohen, Irun R., editor, Lajtha, Abel, editor, Lambris, John D., editor, Paoletti, Rodolfo, editor, and Wilkins, Jon F., editor
- Published
- 2008
- Full Text
- View/download PDF
8. Radiation-Induced Genomic Instability in the Offspring of Irradiated Parents
- Author
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Dubrova, Yuri E., Mothersill, Carmel, editor, Mosse, Irma, editor, and Seymour, Colin, editor
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- 2007
- Full Text
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9. The Function of Toll-Like Receptors
- Author
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Dembic, Zlatko and Rich, Tina
- Published
- 2005
- Full Text
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10. Studies on the reaction in tissue culture of tomato genotypes under biotic stress
- Author
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Ewa Hanus-Fajerska
- Subjects
micropropagation ,morphogenesis ,somatic tissue ,viral infection ,biotic stress ,tomato ,Botany ,QK1-989 - Abstract
Plant regeneration in vitro from virus-infected somatic tomato (Lycopersicon sp.) tissue was performed. Regeneration experiments were started after the determination of virus presence, using enzyme-linked immunosorbent assay, in leaves used as a source of explants. Leaf explants infected with selected strains of tomato mosaic Tobamovirus or cucumber mosaic Cucumovirus respectively, were cultured on a standarised MS agar medium to induce adventitious shoots, which were afterwards excised, rooted in vitro and cultured to plants. Explants were also screened for their ability to produce callus. Diverse effects of viral infection, ranging from stimulation to inhibition of callus formation and of morphogenesis rate, were observed. The health condition of the tissue proved to affect regeneration potential of Lycopersicon esculentum, whereas wild accesions did not react in that case so distinctly. In cultivated tomato was encountered the decline in competence to reproduce shoots adventitiously in infected tissue. There was also relationship between donor plant health condition and adventitious root formation in regenerated shoots. Experiments with short-term cultures of L. esculenum reveled also that a certain number of shoots regenerated from diseased tissue can be virus-free.
- Published
- 2014
- Full Text
- View/download PDF
11. Meiosis was another unexpected property : The cell could reduce its chromosome number
- Author
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Lima-de-Faria, A. and Lima-de-Faria, A.
- Published
- 2003
- Full Text
- View/download PDF
12. Cryostorage of Somatic Tissues of Endangered Australian Species
- Author
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Touchell, D. H., Turner, S. R., Bunn, E., Dixon, K. W., Bajaj, Y. P. S., editor, and Towill, Leigh E., editor
- Published
- 2002
- Full Text
- View/download PDF
13. In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different requirements.
- Author
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Santos, Magda L. T., Borges, Alana A., Queiroz Neta, Luiza B., Santos, Maria V. O., Oliveira, Moacir F., Silva, Alexandre R., and Pereira, Alexsandra F.
- Abstract
Copyright of Pesquisa Veterinaria Brasileira is the property of Colegio Brasileiro de Patologia Animal - CBPA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
- Published
- 2016
- Full Text
- View/download PDF
14. The biochemical composition and calorie density of the walleye pollock Theragra chalcogramma in the Sea of Okhotsk.
- Author
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Gorbatenko, K. and Lazhentsev, A.
- Abstract
In tissues of the walleye pollock Theragra chalcogramma the dry matter content averages 18.5%. The lipid content of the raw material is 0.7%, the protein content is 15.3%, carbohydrates are 0.6%, and ash is 1.3%. The average calorie density is 940 cal/g wet weight and 5080 cal/g dry weight. The dry matter content of gonads varies within 14.9-28.0% in females and 14.5-17.0% in males. The lipid content of the raw material is 0.9-3.0% in females and 1.3-1.8% in males; the protein content is 10.2-21.5% and 10.7-13.4%, respectively. The calorie density of female gonads is 702-1537 cal/g wet weight and 4426-5482 cal/g dry weight; for the male gonads it is 760-960 cal/g wet weight and 4952-5641 cal/g dry weight. The dry matter content of the liver varies within 42.2-62.2% for females and 34.4-62.4 for males. The lipid content of the raw material is 25.6-44.5% for females and 16.6-41.3% for males; the protein content is 6.3-9.8% and 8.1-12.3%, respectively. The calorie density of the liver in females varies within 2918-4601 cal/g wet weight and 6370-7395 cal/g dry weight; in males it is 2291-4357 cal/g wet weight and 6392-7492 cal/g dry weight. The minimum calorie density of the liver is observed in juvenile pollock: 963 cal/g wet weight and 2045 cal/g dry weight. The dry-matter content of feces in different size groups varies within 15.0-18.4%. Values of the average lipid content of raw material range from 1.1 to 1.6%; the protein content is from 1.8 to 3.8% and carbohydrates are from 0.9 to 1.4%. The calorie density of feces from variously-sized walleye pollock varies within a narrow range, from 308 to 362 cal/g wet weight. The energy equivalent ranges, depending on body size, from 259 to 2377 cal. The share of energy concentrated in the somatic (muscle) tissue of variously-sized walleye pollock during ontogenesis constitutes 56.5-93.9%; in female gonads it is 0.9-26.6%; in male gonads it is 0.4-7.3%, in the female liver it is 7.9-27.2%, and in the male liver it is 5.7-26.9%. The amount of energy (cal), concentrated in the female liver and gonads is on average 1.5 and 3 times as high as that in the male liver and gonads, respectively. The maximum total energy loss (15-30%) in mature walleye pollock of various-sizes occurs in the spawning period, during the transition from the maturity stage 5 to stage 6. The total amount of energy accumulated during the lifecycle from small juveniles (<17 cm) to very large individuals (>60 cm) averages 1964 kcal for females and 1465 kcal for males. The difference in the amount of energy is explained by the fact that oogenesis requires more energy than spermatogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
15. Murine β1,4-Galactosyltransferase : Analysis of a Gene That Serves Both A Housekeeping and a Cell Specific Function
- Author
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Shaper, Joel H., Harduin-Lepers, Anne, Rajput, Bhanu, Shaper, Nancy L., Alavi, Azita, editor, and Axford, John S., editor
- Published
- 1995
- Full Text
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16. Gene Transfer via Particle Bombardment: Applications of the Accell Gene Gun
- Author
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Yang, Ning-Sun, De Luna, Carolyn, Cheng, Liang, and Wolff, Jon A., editor
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- 1994
- Full Text
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17. Arabidopsis as a Model System for Analysis of Leaf Senescence and Inflorescence-Meristem Longevity
- Author
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Hensel, Linda L., Bleecker, Anthony B., and Amasino, Richard M., editor
- Published
- 1993
- Full Text
- View/download PDF
18. Reduced representation bisulphite sequencing of ten bovine somatic tissues reveals DNA methylation patterns and their impacts on gene expression.
- Author
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Yang Zhou, Lingyang Xu, Bickhart, Derek M., abdel Hay, El Hamidi, Schroeder, Steven G., Connor, Erin E., Alexander, Leeson J., Sonstegard, Tad S., Van Tassell, Curtis P., Hong Chen, and Liu, George E.
- Subjects
- *
DNA methylation , *SULFITES , *SOMATIC cells , *GENE expression , *CPG nucleotides , *CYTOSINE , *NUCLEOTIDE sequencing - Abstract
Background: As a major epigenetic component, DNA methylation plays important functions in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in economically important animals like cattle. Results: Using reduced representation bisulphite sequencing (RRBS), we obtained single-base-resolution maps of bovine DNA methylation from ten somatic tissues. In total, we evaluated 1,868,049 cytosines in CG-enriched regions. While we found slightly low methylation levels (29.87 to 38.06 %) in cattle, the methylation contexts (CGs and non-CGs) of cattle showed similar methylation patterns to other species. Non-CG methylation was detected but methylation levels in somatic tissues were significantly lower than in pluripotent cells. To study the potential function of the methylation, we detected 10,794 differentially methylated cytosines (DMCs) and 836 differentially methylated CG islands (DMIs). Further analyses in the same tissues revealed many DMCs (including non-CGs) and DMIs, which were highly correlated with the expression of genes involved in tissue development. Conclusions: In summary, our study provides a baseline dataset and essential information for DNA methylation profiles of cattle. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
19. Ancestral TCDD exposure promotes epigenetic transgenerational inheritance of imprinted gene Igf2: Methylation status and DNMTs.
- Author
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Ma, Jing, Chen, Xi, Liu, Yanan, Xie, Qunhui, Sun, Yawen, Chen, Jingshan, Leng, Ling, Yan, Huan, Zhao, Bin, and Tang, Naijun
- Subjects
- *
TETRACHLORODIBENZODIOXIN , *EPIGENETICS , *DNA methyltransferases , *ARYL hydrocarbon receptors , *SOMATOMEDIN , *GENE expression , *POLYMERASE chain reaction , *ANALYSIS of variance - Abstract
Ancestral TCDD exposure could induce epigenetic transgenerational phenotypes, which may be mediated in part by imprinted gene inheritance. The aim of our study was to evaluate the transgenerational effects of ancestral TCDD exposure on the imprinted gene insulin-like growth factor-2 (Igf2) in rat somatic tissue. TCDD was administered daily by oral gavage to groups of F0 pregnant SD rats at dose levels of 0 (control), 200 or 800 ng/kg bw during gestation day 8–14. Animal transgenerational model of ancestral exposure to TCDD was carefully built, avoiding sibling inbreeding. Hepatic Igf2 expression of the TCDD male progeny was decreased concomitantly with hepatic damage and increased activities of serum hepatic enzymes both in the F1 and F3 generation. Imprinted Control Region (ICR) of Igf2 manifested a hypermethylated pattern, whereas methylation status in the Differentially Methylated Region 2 (DMR2) showed a hypomethylated manner in the F1 generation. These epigenetic alterations in these two regions maintained similar trends in the F3 generation. Meanwhile, the expressions of DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) changed in a non-monotonic manner both in the F1 and F3 generation. This study provides evidence that ancestral TCDD exposure may promote epigenetic transgenerational alterations of imprinted gene Igf2 in adult somatic tissue. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
20. Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536
- Author
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Bruno Mezzetti, Silvia Sabbadini, A. Ricci, Luca Capriotti, and O. Navacchi
- Subjects
0106 biological sciences ,0301 basic medicine ,Prunus persica ,somatic tissue ,cefotaxime ,Organogenesis ,Plant Science ,Biology ,01 natural sciences ,STS ,Article ,Petiole (botany) ,03 medical and health sciences ,Prunus ,Basal shoot ,Cultivar ,Ecology, Evolution, Behavior and Systematics ,Ecology ,in vitro organogenesis ,fungi ,Botany ,food and beverages ,Horticulture ,030104 developmental biology ,QK1-989 ,Shoot ,Rootstock ,carbenicillin ,010606 plant biology & botany ,Explant culture - Abstract
In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 &mu, M N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 &mu, M STS or carbenicillin (238 mM) combined with cefotaxime (210 &mu, M) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.
- Published
- 2020
21. Somatic Tissue
- Author
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Schwab, Manfred, editor
- Published
- 2011
- Full Text
- View/download PDF
22. Proteasome dysfunction in Drosophila signals to an Nrf2-dependent regulatory circuit aiming to restore proteostasis and prevent premature aging.
- Author
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Tsakiri, Eleni N., Sykiotis, Gerasimos P., Papassideri, Issidora S., Terpos, Evangelos, Dimopoulos, Meletios A., Gorgoulis, Vassilis G., Bohmann, Dirk, and Trougakos, Ioannis P.
- Subjects
- *
PROTEASOMES , *DENATURATION of proteins , *DROSOPHILA , *PREMATURE aging (Medicine) , *RNA interference , *PHENOTYPES - Abstract
The ubiquitin-proteasome system is central to the regulation of cellular proteostasis. Nevertheless, the impact of in vivo proteasome dysfunction on the proteostasis networks and the aging processes remains poorly understood. We found that RNAi-mediated knockdown of 20 S proteasome subunits in Drosophila melanogaster resulted in larval lethality. We therefore studied the molecular effects of proteasome dysfunction in adult flies by developing a model of dose-dependent pharmacological proteasome inhibition. Impaired proteasome function promoted several 'old-age' phenotypes and markedly reduced flies' lifespan. In young somatic tissues and in gonads of all ages, loss of proteasome activity induced higher expression levels and assembly rates of proteasome subunits. Proteasome dysfunction was signaled to the proteostasis network by reactive oxygen species that originated from malfunctioning mitochondria and triggered an Nrf2-dependent upregulation of the proteasome subunits. RNAi-mediated Nrf2 knockdown reduced proteasome activities, flies' resistance to stress, as well as longevity. Conversely, inducible activation of Nrf2 in transgenic flies upregulated basal proteasome expression and activity independently of age and conferred resistance to proteotoxic stress. Interestingly, prolonged Nrf2 overexpression reduced longevity, indicating that excessive activation of the proteostasis pathways can be detrimental. Our in vivo studies add new knowledge on the proteotoxic stress-related regulation of the proteostasis networks in higher metazoans. Proteasome dysfunction triggers the activation of an Nrf2-dependent tissue- and age-specific regulatory circuit aiming to adjust the cellular proteasome activity according to temporal and/or spatial proteolytic demands. Prolonged deregulation of this proteostasis circuit accelerates aging. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
23. Somatic Tissue
- Author
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Schwab, Manfred, editor
- Published
- 2009
- Full Text
- View/download PDF
24. ASB-1, a germline-specific isoform of mitochondrial ATP synthase b subunit, is required to maintain the rate of germline development in Caenorhabditis elegans
- Author
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Kawasaki, Ichiro, Hanazawa, Momoyo, Gengyo-Ando, Keiko, Mitani, Shohei, Maruyama, Ichiro, and Iino, Yuichi
- Subjects
- *
CAENORHABDITIS elegans , *MITOCHONDRIA , *ADENOSINE triphosphatase , *GERM cells - Abstract
Abstract: The developmental timing of all types of cells must be synchronized and spatially coordinated to achieve the organized development of a multicellular organism. Previously, we found RNAi of asb-1, encoding a germline-specific isoform of mitochondrial ATP synthase b subunit, caused 100% penetrant sterility in Caenorhabditis elegans. ATP synthase is one of the five complexes of the mitochondrial respiratory chain, and defects in some of the components of the chain are known to slow the growth and extend the lifespan of worms. We found that development of asb-1 mutant germ line was not arrested at any stage, but did slow to half the rate of wild type, whereas the rate of somatic development was the same in asb-1 mutants as that of wild type, indicating that asb-1 is required to maintain the rate of germline development but has no effect on somatic development. Among ATP synthase subunit genes, RNAi of asg-1, encoding a germline-specific isoform of the g subunit, also caused asb-1-like sterility, indicating that some other germline-specific components are also required to maintain the rate of germline development. Both asb-1 and asg-1 are located on autosomes while they possess counterparts, asb-2 and asg-2, respectively, on X chromosome, which are both required for somatic development. Chromosomal locations of the genes may be the basis of the segregation of germline/somatic functions of each gene, as were demonstrated for other autosomal/X-linked duplicated gene pairs. [Copyright &y& Elsevier]
- Published
- 2007
- Full Text
- View/download PDF
25. In vitro plant regeneration from somatic tissue of strawberry Fragaria x ananassa Duch.
- Author
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Jasna Berljak, Mojca Marn, and Darinka Koron
- Subjects
in vitro ,somatic tissue ,leax explants ,plant regenration ,strawberry ,cv. Elsanta ,Biology (General) ,QH301-705.5 - Abstract
Successful shoot regeneration in somatic tissue is the basic requirement for in vitro induction of genetic variability as the new tool in plant breeding. Somatic tissue excised from in vitro multiplied strawberry plants were tested on ability for plant regeneration. Leaves, petiole and stipules were inoculated on initial medium with BA and 2,4-D, or on medium with BA only after 1 hour pulse treatment with 2,4-D. Callus was induced on all sliced surfaces of explants inoculated on initial medium with growth regulators BA and 2,4-D during first 7 days of culture. Explants inoculated on initial medium with BA, after pulse treatment with 2,4-D did not develop callus but abundantly produced fenolic compounds, and tured necrotic in the first 24 hours. Spontaneous plant regeneration was noticed on leaves explants with less developed callus tissue on initial medium with growth regulators duringsecond week of culture. High percentage of explants with regenerated shoots were obtained after transfer on hormone-free medium. The highest plant regeneration ability was in leaf tissue, less in petiole and stipules. Callus induced in leaf tissue showed ability for constant plant regeneration during three months of culture and careful 4-week interval transfer on basal MS medium with 4.4¼M BA and 40 g/l sucrose.
- Published
- 2003
- Full Text
- View/download PDF
26. Tissue distribution and prevalence of Wolbachia infections in tsetse flies, Glossina spp.
- Author
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Cheng, Q., Ruel, T. D., Zhou, W., Moloo, S. K., Majiwa, P., O'neill, S. L., and Aksoy, S.
- Subjects
- *
TSETSE-flies , *INFECTION - Abstract
SummaryTsetse flies Glossina spp. (Diptera: Glossinidae) harbor three different symbiotic microorganisms, one being an intracellular Rickettsia of the genus Wolbachia. This bacterium infects a wide range of arthropods, where it causes a variety of reproductive abnormalities, one of which is termed cytoplasmic incompatibility (CI) that, when expressed, results in embryonic death due to disruptions in fertilization events. We report here that in colonized flies, Wolbachia infections can be detected in 100% of sampled individuals, while infections vary significantly in field populations. Based on Wolbachia Surface Protein (wsp) gene sequence analysis, the infections associated with different fly species are all unique within the A group of the Wolbachia pipientis clade. In addition to being present in germ-line tissues, Wolbachia infections have been found in somatic tissues of several insects. Using a Wolbachia-specific PCR-based assay, the tissue tropism of infections in Glossina morsitans morsitans Westwood, Glossina brevipalpis Newstead and Glossina austeni Newstead were analysed. While infections in G. m. morsitans and G. brevipalpis were limited to reproductive tissues, in G. austeni, Wolbachia could be detected in various somatic tissues. [ABSTRACT FROM AUTHOR]
- Published
- 2000
- Full Text
- View/download PDF
27. EVALUATION OF QUALITY INDICES OF THE GONAD AND SOMATIC TISSUES INVOLVED IN REPRODUCTION OF THE PEARL OYSTER PINCTADA MAZATLANICA WITH HISTOCHEMISTRY AND DIGITAL IMAGE ANALYSIS.
- Author
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Gómez-Robles, Eliana and Saucedo, Pedro E.
- Abstract
We evaluated indicators of quality for female gonads (ovary and oocytes), male gonads (testis and seminal tubules), and selected somatic tissues (fiber packages in the adductor muscle and adenomeres in the digestive gland) that participate in reproduction of the pearl oyster Pinctada mazatlanica. The goal was to identify timing of optimal broodstock condition for larval rearing practices. Tissue samples were collected seasonally and processed with a combination of histochemistry and digital image analysis to develop a gonad tissue index, a lipid index, and a glycogen index. Seasonal changes in these indicators were correlated with changes in water temperature and chlorophyll a in the water. P. mazatlanica uses a combination of stored reserves and food supply (conservative vs. opportunistic strategy) to regulate reproduction, but the way energy is acquired and allocated varies between sexes. Female gonads contained higher lipid contents during spring. We suggest energy allocation from digestive gland, because this tissue showed lower lipid contents in the same season (conservative strategy). Within oocytes, the accumulation of lipids occurred from nutrients obtained from food supply during winter (19.6°C and ~650 ng/L) (opportunistic strategy). Male gonads contained higher glycogen contents in spring. A decreasing trend in the glycogen content of the adductor muscle was also detected in spring. This suggests that sperm build-up occurs partly at the expense of the glycogen stored in this tissue (conservative strategy). In seminal tubules, no correlation with the glycogen content of adductor muscle was detected, suggesting that these reserves were obtained from food supply in spring (22°C and ~400 ng/L) (opportunistic strategy). Optimal broodstock condition occurs mainly in spring and secondly in early winter. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
28. Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea)
- Author
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Sabiha Khatoon, Aamir Sohail, Shabnam Khan, Sakil Ahmed, and M.K. Saifullah
- Subjects
0301 basic medicine ,somatic tissue ,purification ,Veterinary medicine ,Size-exclusion chromatography ,SF1-1100 ,03 medical and health sciences ,chemistry.chemical_compound ,SF600-1100 ,Parasite hosting ,Polyacrylamide gel electrophoresis ,Ammonium sulfate precipitation ,glutathione S-transferase ,chemistry.chemical_classification ,Chromatography ,General Veterinary ,biology ,Glutathione ,Enzyme assay ,Animal culture ,Bubalus bubalis ,Gastrothylax crumenifer ,030104 developmental biology ,Enzyme ,Glutathione S-transferase ,chemistry ,biology.protein ,Research Article - Abstract
Aim: Aim of the present study was to carry out the partial purification and biochemical characterization of glutathione S-transferase (GST) from the somatic tissue of ruminal amphistome parasite, Gastrothylax crumenifer (Gc) infecting Indian water buffalo (Bubalus bubalis). Materials and Methods: The crude somatic homogenate of Gc was subjected to progressive ammonium sulfate precipitation followed by size exclusion chromatography in a Sephacryl S 100-HR column. The partially purified GST was assayed spectrophotometrically, and the corresponding enzyme activity was also recorded in polyacrylamide gel. GST isolated from the amphistome parasite was also exposed to variable changes in temperature and the pH gradient of the assay mixture. Results: The precipitated amphistome GST molecules showed maximum activity in the sixth elution fraction. The GST subunit appeared as a single band in the reducing polyacrylamide gel electrophoresis with an apparent molecular weight of 26 kDa. The GST proteins were found to be fairly stable up to 37°C, beyond this the activity got heavily impaired. Further, the GST obtained showed a pH optima of 7.5. Conclusion: Present findings showed that GST from Gc could be conveniently purified using gel filtration chromatography. The purified enzyme showed maximum stability and activity at 4°C.
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- 2014
29. Histological characterization and vitrification of somatic tissue derived from collared peccaries (Pecari tajacu Linnaeus, 1758)
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Borges, Alana Azevedo, Pereira, Alexsandra Fernandes, Silva, Alexandre Rodrigues, and Teixeira, Dárcio Ítalo Alves
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Tecido somático ,CIENCIAS AGRARIAS::MEDICINA VETERINARIA [CNPQ] ,Animais silvestres ,CIENCIAS AGRARIAS [CNPQ] ,crioprotetor ,Somatic tissue ,Cryoprotectant ,Wild animals - Abstract
Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-03-22T14:51:38Z No. of bitstreams: 1 AlanaAB_DISSERT.pdf: 4740716 bytes, checksum: e3666ea7a25906742164df387e99208b (MD5) Made available in DSpace on 2017-03-22T14:51:38Z (GMT). No. of bitstreams: 1 AlanaAB_DISSERT.pdf: 4740716 bytes, checksum: e3666ea7a25906742164df387e99208b (MD5) Previous issue date: 2016-11-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior The cryopreservation of somatic tissue derived from collared peccaries represents an interesting step in the obtainment and conservation of cells for nuclear transfer (cloning). In this sense, tissue vitrification protocols need to be optimized to ensure maximum preservation of viable cell characteristics. Therefore, the aims of this study were to characterize histologically the peripheral auricular integumentary system (Stage 1) and evaluate different cryoprotectants in the solid-surface vitrification of somatic tissue of collared peccaries (Stage 2). Thun, ear fragments (9.0 mm3) were collected of sixteen animals derived from the Multiplication Center of Wild Animals (CEMAS/UFERSA). In the first stage, tissue samples were evaluated for the characterization of layers, its components and proliferative activity. For the second stage, tissue fragments were cryopreserved by solidsurface vitrification using Dulbecco modified minimum essential medium supplemented with 10% fetal bovine serum and different cryoprotectants and concentrations [dimethylsulfoxide (DMSO, 3.0 M), ethylene glycol (EG; 3.0 M) and association DMSO/EG (1.5 M; 1.5 M) in the absence and presence of sucrose (0.25M)]. After two weeks, warmed and non-vitrified (control) fragments were analyzed using histological techniques. Thus, for both steps, tissue samples were evaluated using hematoxylin-eosin and Gomori Trichrome, quantification of regions argyrophilic nucleolar organizer (AgNORs) and viability by MTT assay (3-[4,5- dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). Also, in the first stage, fragments were analyzed by transmission electronic microscopy. Thus, in the first stage, sizes of 104.2 μm and 222.6 μm were observed in the epidermis and dermis, with a volumetric ratio of 36.6% and 58.7%, respectively. Moreover, in the epidermis were evidenced the basal layer (22.5 μm), intermediate (53.5 μm) and cornea (28.2 μm), with mean values of 65.3 epithelial cells, 43.4 melanocytes and 14.8 perinuclear halos. Already the dermis has 127 fibroblasts with 2.5 AgNORs by nucleolus. Additionally, the metabolic activity was 0.243. In the second stage, the 3.0 M EG with sucrose was able to maintain normal tissue characteristics compared with non-vitrified (control), especially for the volumetric ratio of epidermis (61.2 vs. 58.7) and dermis (34.5 vs. 36.6), number of fibroblast (90.3 vs. 127.0), and AgNOR ratio (0.09 vs. 0.17), respectively. In conclusion, the peripheral auricular integumentary system derived from collared peccaries possessed some variations compared to other mammals, as the number of layers and thickness of the epidermis, number of epithelial cells, melanocytes and proliferative parameters. Moreover, 3.0 M EG with 0.25 M sucrose resulted in a better cryoprotectant composition in the vitritification for somatic tissue of collared peccaries A criopreservação de tecido somático de catetos representa uma etapa interessante na obtenção e conservação de células para a transferência nuclear (clonagem). Nesse sentido, protocolos de vitrificação tecidual necessitam ser otimizados para garantir a máxima conservação das características viáveis das células. Portanto, os objetivos do presente trabalho foram caracterizar histologicamente o sistema tegumentar da região auricular periférica (Etapa 1) e avaliar diferentes crioprotetores na vitrificação em superfície sólida de tecido somático de catetos (Etapa 2). Para tanto, fragmentos auriculares (9,0 mm3) foram recuperados de dezesseis animais oriundos do Centro de Multiplicação de Animais Silvestres (CEMAS/UFERSA). Na primeira etapa, amostras teciduais foram avaliadas quanto à caracterização das camadas, seus componentes e atividade proliferativa. Para a segunda etapa, fragmentos teciduais foram criopreservados por vitrificação em superfície sólida em meio essencial mínimo modificado por Dulbecco suplementado com 10% de soro fetal bovino e diferentes crioprotetores e concentrações [dimetilsulfóxido (DMSO; 3,0 M), etilenoglicol (EG; 3,0 M) e associação DMSO/EG (1,5 M; 1,5 M) na ausência e presença de sacarose (0,25 M)]. Após duas semanas, fragmentos aquecidos e não criopreservados (controle) foram analisados usando técnicas histológicas. Assim, para ambas as etapas, amostras teciduais foram avaliadas usando colorações hematoxilina-eosina e tricômico de Gomori, quantificação de regiões argirofílicas organizadoras nucleolares (AgNORs) e viabilidade pelo ensaio de MTT (brometo de 3-[4,5-dimetil-tiazol-2-il]-2,5-difeniltetrazólio). Ainda, na primeira etapa, fragmentos foram analisados por microscopia eletrônica de transmissão. Assim, na primeira etapa, tamanhos de 104,2 μm e 222,6 μm foram observados para epiderme e derme, com uma proporção volumétrica de 36,6% e 58,7%, respectivamente. Além disso, na epiderme, foram evidenciadas as camadas basal (22,5 μm), intermediárias (53,5 μm) e córnea (28,2 μm), com valores médios de 65,3 de células epidermais, 43,4 melanócitos e 14,8 de halos perinucleares. Já a derme apresentou 127 fibroblastos com 2,5 AgNORs por nucléolo. Adicionalmente, a atividade metabólica foi de 0,243. Na segunda etapa, a combinação de 3,0 M de EG com sacarose foi adequada em manter as características teciduais normais quando comparado com os fragmentos não vitrificados, especialmente para a proporção volumétrica da epiderme (61,2 vs. 58,7) e derme (34,5 vs. 36,6), número de fibroblastos (90,3 vs. 127,0) e razão de AgNOR (0,09 vs. 0,17), respectivamente. Em conclusão, o sistema tegumentar auricular periférico de catetos possuiu algumas variações em relação a outros mamíferos, quanto ao número de camadas e espessura da epiderme, quantidade de células epidermais, melanócitos e parâmetros proliferativos. Além disso, 3,0 M de EG com 0,25 M de sacarose resultaram na melhor composição de crioprotetores na vitrificação de tecido somático de catetos 2017-03-21
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- 2016
30. Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536.
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Ricci, Angela, Capriotti, Luca, Mezzetti, Bruno, Navacchi, Oriano, and Sabbadini, Silvia
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PLANT shoots ,PEACH ,ROOTSTOCKS ,WOODY plants ,PLANT regulators ,GENETIC transformation ,PRUNUS - Abstract
In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N
6 -benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars. [ABSTRACT FROM AUTHOR]- Published
- 2020
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31. A Comparative Study of the Diversity of Gene Expression in Brain
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Kaplan, B. B., Gioio, A. E., Capano, C. Perrone, Giuditta, A., Giuditta, Antonio, editor, Kaplan, Barry B., editor, and Zomzely-Neurath, Claire, editor
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- 1986
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32. A Comparison of Genotoxic Activity in Somatic Tissue and in Germ Cells of Drosophila Melanogaster
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Vogel, Ekkehart W., Chu, Ernest H. Y., editor, and Generoso, Walderico M., editor
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- 1984
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33. Ontogenetic patterns and phylogenetic trends in freshwater flatworms (Tricladida); constraint or selection?
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Calow, P., Read, D. A., Dumont, H. J., editor, and Tyler, Seth, editor
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- 1986
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34. The Use of Gaspé Variety for the Study of Pollen and Anther Development in Maize
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Hourcade, Dennis E., Bugg, Michael, Loussaert, Dale F., Mulcahy, David L., editor, Mulcahy, Gabriella Bergamini, editor, and Ottaviano, Ercole, editor
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- 1986
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35. Identification of Duplicate Loci and Evidence for Post-meiotic Gene Expression in Pollen
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Weeden, N. F., Mulcahy, David L., editor, Mulcahy, Gabriella Bergamini, editor, and Ottaviano, Ercole, editor
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- 1986
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36. In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different supplements
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Santos, Magda Lorena Turbano dos, Pereira, Alexsandra Fernandes, Silva, Alexandre Rodrigues, and Freitas, Vicente José de Figueirêdo
- Subjects
Tecido somático ,Protein source ,Conservação ,CIENCIAS AGRARIAS::MEDICINA VETERINARIA [CNPQ] ,CIENCIAS AGRARIAS [CNPQ] ,Animais silvestres ,Fonte proteica ,Fator mitótico ,Mitotic factor ,Somatic tissue ,Conservation ,Wild animals - Abstract
Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-03-07T12:56:11Z No. of bitstreams: 1 MagdaLTS_DISSERT.pdf: 1454861 bytes, checksum: 4612469437c8dd924c24af03a052758d (MD5) Made available in DSpace on 2017-03-07T12:56:11Z (GMT). No. of bitstreams: 1 MagdaLTS_DISSERT.pdf: 1454861 bytes, checksum: 4612469437c8dd924c24af03a052758d (MD5) Previous issue date: 2016-03-30 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior The maintenance of metabolic activities during the in vitro culture of somatic cells of wild animals, especially collared peccary (Pecari tajacu), is an interesting step in the conservation of these cells for use in nuclear transfer (cloning). In this context, it is necessary to optimize the in vitro culture conditions of somatic cell by establishment of some appropriate supplementations to the media, in order to ensure the maximum preservation of the viable cell characteristics. Therefore, this study aimed to optimize the composition of the culture means of somatic cell derived from ear tissue of collared peccaries, evaluating two concentrations of fetal bovine serum (E1: FBS; 10% vs. 20%) and epidermal growth factor (E2: EGF, 5 ng/mL vs. 10 ng/mL). Thus, tissue fragments from 18 adult animals were submitted to primary culture and subcultures for 40 days until the fourth passage and the resulting cells were analyzed for morphology, adhesion, subconfluence, proliferative activity for developing growth curve for seven days and determining the population doubling time (PDT), viability by trypan blue and functional/metabolic activity by assay 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). Moreover, in the E1, comparisons as cell adhesion were performed with cells cultured in the presence of bovine serum albumin (BSA, 0.5% and 1.0%). All data were analyzed by ANOVA followed by post hoc test. In the E1, no difference (P>0.05) was observed between the concentrations of FBS for the number of adhered [SFB10: 39/39 vs. SB20: 35/39] and subconfluent fragments [SFB10: 39/39 vs. SB20: 35/39], day all adhered [SFB10: 3.5 vs. SFB20: 3.0], with growth [SFB10: 7.4 vs. SFB20: 7.2] and subconfluent samples [SFB10: 11.8 vs. SFB20: 11.8]. However, significant values were observed in cells cultured in the presence of 20% FBS for viability [SFB10: 85.6% vs. SFB20: 98.2%], PDT [SFB10: 155.4 h vs.77.25 h] and MTT assay [SFB10: 0.57-0.57 vs. SFB20: 0.82-0.99 (D5-D7)]. Additionally, comparisons of supplementation of BSA and FBS confirm the potential FBS cell adhesion. Thus, 20% FBS was used in the following experiment. In the evaluation of the presence of EGF in culture, no difference was observed in the evaluated parameters for the number of attached [EGF0: 31/31 vs. EGF5: 31/31 vs. EGF10: 31/31] and subconfluent fragments [EGF0: 31/31 vs. EGF5: 31/31 vs. EGF10: 31/31] day all adhered [EGF0: 4.9 vs. EGF5: 7.0 vs. EGF10: 3.5] growth [EGF0: 7.2 vs. EGF5: 8.2 vs. EGF10: 7.9] and subconfluent samples [0 EGF: 12.6 vs. EGF5: 16.6 vs. EGF10:12.6], viability [EGF0: 84.3% vs. EGF5: 88.8% vs. EGF10: 87.0%], PDT [EGF0: 69.6 h vs. EGF5: 64.8 h vs. EGF10: 65.3 h] and MTT assay [EGF0: 1.26-1.38 vs. EGF5: 1.06-1.14 vs. EGF10: 1.13-1.16 (D5-D7)]. In all experiments, the growth curves showed clear log and lag phases of development. In conclusion, 20% FBS is suitable for the recovery of somatic cells in vitro; however, EGF does not improve the quality of growing these cells A manutenção das atividades metabólicas durante o cultivo in vitro de células somáticas de animais silvestres, especialmente catetos (Pecari tajacu), representa uma etapa interessante na conservação dessas células para aplicação na transferência nuclear (clonagem). Nesse contexto, faz-se necessário a otimização das condições de cultivo in vitro de células somáticas pelo estabelecimento de algumas suplementações adequadas aos meios, visando garantir a máxima preservação das características celulares viáveis. Portanto, o presente trabalho teve como objetivo otimizar a composição do meio de cultivo de células somáticas derivadas de tecido auricular de catetos, avaliando duas concentrações de soro fetal bovino (E1: SFB; 10% vs. 20%) e fator de crescimento epidermal (E2: EGF; 5 ng/mL vs. 10 ng/mL). Para tanto, fragmentos teciduais de 18 animais adultos foram submetidos ao cultivo primário e subcultivos por 40 dias, até a quarta passagem e as células resultantes foram analisadas quanto à morfologia, aderência, subconfluência, atividade proliferativa pela elaboração de curva de crescimento por sete dias e determinação do tempo de duplicação da população (PDT), viabilidade por azul de tripano e atividade funcional/metabólica pelo ensaio de brometo de 3-[4,5-dimetil-tiazol-2-il]-2,5-difeniltetrazólio (MTT). Além disso, no E1, comparações quanto à adesão celular foram realizadas com células cultivadas na presença de albumina sérica bovina (BSA, 0,5% e 1,0%). Todos os dados foram analisados por ANOVA seguido por teste post-hoc. No E1, nenhuma diferença (P>0,05) foi observada entre as concentrações de SFB para o número de fragmentos aderidos [SFB10: 39/39 vs. SB20: 35/39] e subconfluentes [SFB10: 39/39 vs. SB20: 35/39], dia de todas as amostras aderidas [SFB10: 3,5 vs. SFB20: 3,0], com crescimento [SFB10: 7,4 vs. SFB20: 7,2] e subconfluentes [SFB10: 11,8 vs. SFB20: 11,8]. Contudo, valores significativos foram observados em células cultivadas na presença de SFB a 20% quanto à viabilidade [SFB10: 85,6% vs. SFB20: 98,2%], PDT [SFB10: 155,4 h vs.77,2 h] e ensaio de MTT [SFB10: 0,57-0,57 vs. SFB20: 0,82-0,99 (D5-D7)]. Adicionalmente, comparações da suplementação do BSA e SFB confirmaram o potencial de adesão celular do soro. Assim, SFB a 20% foi empregado no experimento seguinte. Já na avaliação da presença de EGF no cultivo, nenhuma diferença foi observada nos parâmetros avaliados para o número de fragmentos aderidos [EGF0: 31/31 vs. EGF5: 31/31 vs. EGF10: 31/31] e subconfluentes [EGF0: 31/31 vs. EGF5: 31/31 vs. EGF10: 31/31], dia de todas as amostras aderidas [EGF0: 4,9 vs. EGF5: 7,0 vs. EGF10: 3,5], em crescimento [EGF0: 7,2 vs. EGF5: 8,2 vs. EGF10: 7,9] e subconfluentes [EGF0: 12,6 vs. EGF5: 16,6 vs. EGF10: 12,6], viabilidade [EGF0: 84,3% vs. EGF5: 88,8% vs. EGF10: 87,0%], PDT [EGF0: 69,6 h vs. EGF5: 64,8 h vs. EGF10: 65,3 h] e ensaio de MTT [EGF0: 1,26-1,38 vs. EGF5: 1,06-1,14 vs. EGF10: 1,13-1,16 (D5-D7)]. Em todos os experimentos, as curvas de crescimento apresentaram nítidas fases log e lag de desenvolvimento. Em conclusão, o SFB a 20% é adequado para a recuperação de células somáticas in vitro; contudo, o EGF não melhora a qualidade do cultivo dessas células 2017-03-07
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- 2016
37. Material and Methods
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Offerijns, Frederik Jan Maurits and Offerijns, Frederik Jan Maurits
- Published
- 1935
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38. Development of DNA delivery system using Pseudomonas exotoxin A and a DNA binding region of human DNA topoisomerase I
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Chen, T.-Y., Hsu, C.-T., Chang, K.-H., Ting, C.-Y., Whang-Peng, J., Hui, C.-F., and Hwang, J.
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- 2000
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39. In vitro culture of somatic cells derived from ear tissue of collared peccary (Pecari tajacu Linnaeus, 1758) in medium with different requirements
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Magda L.T. Santos, Alana A. Borges, Luiza B. Queiroz Neta, Maria V.O. Santos, Moacir F. Oliveira, Alexandre R. Silva, and Alexsandra F. Pereira
- Subjects
Somatic cells ,collared peccary ,Pecari tajacu ,culture medium ,wild animals ,conservation ,somatic tissue ,protein source ,mitotic factor. ,Veterinary medicine ,SF600-1100 - Abstract
ABSTRACT: The maintenance of metabolic activities during the in vitro culture of somatic cells of wild animals, especially collared peccary (Pecari tajacu), is an interesting step in conservation of these cells for the use in nuclear transfer. In this context, it is necessary to optimize the culture conditions of somatic cells by the establishment of appropriate supplementation to the media. Therefore, this study aimed to analyze the composition of the culture means of somatic cell derived from ear tissue of collared peccaries, evaluating concentrations of fetal bovine serum (FBS; 10% vs. 20%) and epidermal growth factor (EGF; 5ng/mL vs. 10ng/mL). Tissues were submitted to primary culture and subcultures for 40 days and cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity to develop the growth curve and to determine the population doubling time (PDT), viability, and functional/metabolic activity. No difference was observed between the concentrations of FBS for several parameters, except for viability [FBS10: 85.6% vs. FBS20: 98.2%], PDT [FBS10: 155.4h vs. 77.2h], and functional/metabolic assay [FBS10: 0.57-0.55 vs. FBS20: 0.82-0.99 (D5-D7)]. For the EGF in culture, no difference was observed in the evaluated parameters. In all experiments, the growth curves were typical S-shape and the cells passed through a lag, logarithmic, and plateau phase. In conclusion, 20% FBS is suitable for the recovery of somatic cells; nevertheless, EGF does not improve the quality of growing these cells. To our knowledge, this is the first study culturing somatic cells of collared peccaries.
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40. Genomic analysis of the mouse protamine 1, protamine 2, and transition protein 2 gene cluster reveals hypermethylation in expressing cells
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Choi, Young-Chul, Aizawa, Akira, and Hecht, Norman B.
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- 1997
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41. CpG content affects gene silencing in mice: evidence from novel transgenes
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Chevalier, Christine, Henry, Isabelle, Montfort, Lucile, Capgras, Suzanne, Forlani, Sylvie, Muschler, John, Nicolas, Jean-François, Biologie Moléculaire du Développement, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Vividis Biotech [Canada], University of California [Berkeley], University of California, This work was supported by grants from ARC (Association pour la Recherche sur le Cancer), CNRS (Centre National pour la Recherche Scientifique) and AFM (Association Française contre les Myopathies). C.C.M. is a student of EpHE (Ecole pratique des Hautes Etudes), J.F.N. and I.H. are from the Institut National de la Recherche Médicale (INSERM)., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), University of California [Berkeley] (UC Berkeley), and University of California (UC)
- Subjects
Male ,MESH: Mutation ,MESH: Mice, Transgenic ,[SDV]Life Sciences [q-bio] ,Mice, Transgenic ,MESH: Transgenes ,Mice ,Peptide Elongation Factor 1 ,MESH: DNA Methylation ,MESH: Mice, Inbred C57BL ,MESH: Plasmids ,MESH: Gene Expression Regulation, Developmental ,MESH: Promoter Regions, Genetic ,Animals ,MESH: Animals ,MESH: Gene Silencing ,Gene Silencing ,Transgenes ,MESH: Peptide Elongation Factor 1 ,Promoter Regions, Genetic ,MESH: CpG Islands ,MESH: Mice ,MESH: Mutagenesis ,MESH: Mice, Inbred DBA ,Research ,MESH: Spermatozoa ,LacZ Reporter ,MESH: Embryo, Mammalian ,Gene Expression Regulation, Developmental ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Methylation Pattern ,MESH: Lac Operon ,DNA Methylation ,Epigenetic Control ,Embryo, Mammalian ,Spermatozoa ,Somatic Tissue ,MESH: Male ,Mice, Inbred C57BL ,Lac Operon ,Mice, Inbred DBA ,Mutagenesis ,Mutation ,Male Germ Line ,CpG Islands ,Female ,MESH: Female ,Plasmids - Abstract
It has been demonstrated that the density of CpG sequences in the transcribed regions of transgenes can have a causal role in repression of transcription. These results show that the mechanism by which CpG islands escape de novo methylation is sensitive to CpG density of adjacent sequences., Background Transgenes are often engineered using regulatory elements from distantly related genomes. Although correct expression patterns are frequently achieved even in transgenic mice, inappropriate expression, especially with promoters of widely expressed genes, has been reported. DNA methylation has been implicated in the aberrant expression, but the mechanism by which the methylation of a CpG-rich sequence can perturb the functioning of a promoter is unknown. Results We describe a novel method for analyzing epigenetic controls that allows direct testing of CpGs involvement by using LacZ reporter genes with a CpG content varying from high to zero that are combined with a CpG island-containing promoter of a widely expressed gene - the α-subunit of the translation elongation factor 1. Our data revealed that a LacZ transgene with null CpG content abolished the strong transgene repression observed in the somatic tissues of transgenic lines with higher CpG content. Investigation of transgene expression and methylation patterns suggests that during de novo methylation of the genome the CpG island-containing promoter escapes methylation only when combined with the CpG-null transgene. In the other transgenic lines, methylation of the promoter may have led to transcriptional silencing. Conclusions We demonstrate that the density of CpG sequences in the transcribed regions of transgenes can have a causal role in repression of transcription. These results show that the mechanism by which CpG islands escape de novo methylation is sensitive to CpG density of adjacent sequences. These findings are of importance for the design of transgenes for controlled expression.
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- 2003
42. Cryopreservation of somatic germplasm of selected Australian monocotyledonous taxa (Haemodoraceae).
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Turner, Shane and Turner, Shane
- Abstract
The South West Botanical Province of Western Australia is one of the most floristically rich areas of the world with over, 8,000 species present, the majority of which (70%), are endemic to this region. Coupled with this high level of endemism, many taxa are threatened which makes them vulnerable to habitat alterations, modifications and destruction. Significant habitat alteration in many areas has resulted in 27 species becoming extinct in the South West Botanical Province, while an additional 327 species are classified as rare and endangered. In the context of stemming this loss of biodiversity, research in cryopreservation was undertaken to provide offsite protection and conservation of somatic germplasm.Cryostorage techniques were evaluated in this study to determine the key factors which may affect the ability of somatic tissues of Haemodoraceae species to survive, recover and grow following liquid nitrogen (LN) immersion and storage. Using Anigozanthos viridis as a comparator in most experiments, the base vitrification protocol was established, which involved: (1) preculturing shoot apices on 0.4 M sorbitol for 48 h; (2) incubation in a vitrification cryoprotective solution (PVS2) for 25 min at 0 degrees celsius; (3) LN immersion; (4) recovery to active growth through warming (immersion in a 40 degrees celsius water bath). Using this procedure the highest post-LN survival of shoot apices for A. viridis was 41.4 plus or minus 6.1% Four additional taxa were successfully cryopreserved with this base protocol (Anigozanthos manglesii, A. rufus, Conostylis wonganensis and A. rufus x A. pulcherrimus); a fifth taxon, Macropidia fuliginosa, however, proved unresponsive.To improve on post-LN survival, further research established that four of the six study taxa responded to the following amendments to the basic protocol: (1) longer preculture duration; (2) preculture on 0.8 M glycerol rather than sorbitol, (3) utilisation of PVS2 solutions with reduced DMSO content; and
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- 2001
43. Reduced representation bisulphite sequencing of ten bovine somatic tissues reveals DNA methylation patterns and their impacts on gene expression.
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Zhou Y, Xu L, Bickhart DM, Abdel Hay EH, Schroeder SG, Connor EE, Alexander LJ, Sonstegard TS, Van Tassell CP, Chen H, and Liu GE
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- Animals, Cattle, CpG Islands, Epigenesis, Genetic, Epigenomics methods, Organ Specificity genetics, Sequence Analysis, DNA, DNA Methylation, Gene Expression
- Abstract
Background: As a major epigenetic component, DNA methylation plays important functions in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in economically important animals like cattle., Results: Using reduced representation bisulphite sequencing (RRBS), we obtained single-base-resolution maps of bovine DNA methylation from ten somatic tissues. In total, we evaluated 1,868,049 cytosines in CG-enriched regions. While we found slightly low methylation levels (29.87 to 38.06 %) in cattle, the methylation contexts (CGs and non-CGs) of cattle showed similar methylation patterns to other species. Non-CG methylation was detected but methylation levels in somatic tissues were significantly lower than in pluripotent cells. To study the potential function of the methylation, we detected 10,794 differentially methylated cytosines (DMCs) and 836 differentially methylated CG islands (DMIs). Further analyses in the same tissues revealed many DMCs (including non-CGs) and DMIs, which were highly correlated with the expression of genes involved in tissue development., Conclusions: In summary, our study provides a baseline dataset and essential information for DNA methylation profiles of cattle.
- Published
- 2016
- Full Text
- View/download PDF
44. Cultivo in vitro de células somáticas derivadas de tecido auricular de cateto (Pecari tajacu Linnaeus, 1758) em meio com diferentes requerimentos
- Author
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Alana Azevedo Borges, Luiza Bento de Queiroz Neta, Moacir Franco de Oliveira, Alexandre Rodrigues Silva, M. L. T. Santos, Alexsandra Fernandes Pereira, and María Victoria Santos
- Subjects
0301 basic medicine ,Pecari ,Pecari tajacu ,culture medium ,somatic tissue ,Somatic cell ,fator mitótico ,Context (language use) ,Biology ,mitotic factor ,Andrology ,03 medical and health sciences ,tecido somático ,Epidermal growth factor ,wild animals ,tecido auricular ,Doubling time ,conservação ,Somatic cells ,protein source ,fonte proteica ,lcsh:Veterinary medicine ,General Veterinary ,meio de cultivo ,0402 animal and dairy science ,conservation ,collared peccary ,04 agricultural and veterinary sciences ,Growth curve (biology) ,biology.organism_classification ,040201 dairy & animal science ,In vitro ,animais silvestres ,030104 developmental biology ,catetos ,Células somáticas ,Immunology ,lcsh:SF600-1100 ,Fetal bovine serum - Abstract
The maintenance of metabolic activities during the in vitro culture of somatic cells of wild animals, especially collared peccary (Pecari tajacu), is an interesting step in conservation of these cells for the use in nuclear transfer. In this context, it is necessary to optimize the culture conditions of somatic cells by the establishment of appropriate supplementation to the media. Therefore, this study aimed to analyze the composition of the culture means of somatic cell derived from ear tissue of collared peccaries, evaluating concentrations of fetal bovine serum (FBS; 10% vs. 20%) and epidermal growth factor (EGF; 5ng/mL vs. 10ng/mL). Tissues were submitted to primary culture and subcultures for 40 days and cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity to develop the growth curve and to determine the population doubling time (PDT), viability, and functional/metabolic activity. No difference was observed between the concentrations of FBS for several parameters, except for viability [FBS10: 85.6% vs. FBS20: 98.2%], PDT [FBS10: 155.4h vs. 77.2h], and functional/metabolic assay [FBS10: 0.57-0.55 vs. FBS20: 0.82-0.99 (D5-D7)]. For the EGF in culture, no difference was observed in the evaluated parameters. In all experiments, the growth curves were typical S-shape and the cells passed through a lag, logarithmic, and plateau phase. In conclusion, 20% FBS is suitable for the recovery of somatic cells; nevertheless, EGF does not improve the quality of growing these cells. To our knowledge, this is the first study culturing somatic cells of collared peccaries. RESUMO: A manutenção das atividades metabólicas durante o cultivo in vitro de células somáticas de animais silvestres, especialmente cateto (Pecari tajacu), é uma etapa interessante na conservação dessas células para o uso na transferência nuclear. Nesse contexto, é necessário aperfeiçoar as condições de cultivo de células somáticas pelo estabelecimento de suplementações apropriadas aos meios. Portanto, este estudo objetivou analisar a composição dos meios de cultivo de células somáticas derivadas de tecido auricular de catetos, avaliando concentrações de soro fetal bovino (SFB; 10% vs. 20%) e fator de crescimento epidermal (EGF; 5 ng/mL vs. 10 ng/mL). Para tanto, tecidos foram submetidos ao cultivo primário e subcultivos por 40 dias e células foram analisadas por morfologia, adesão, subconfluência, e atividade proliferativa pelo desenvolvimento da curva de crescimento e determinação do time de duplicação da população (PDT), viabilidade, e atividade funcional/metabólica. Nenhuma diferença foi observada entre as concentrações de SFB para os vários parâmetros, exceto para viabilidade [SFB10: 85,6% vs. SFB20: 98,2%], PDT [SFB10: 155,4 h vs. 77,2 h], e atividade funcional/metabólica [SFB10: 0,57-0,55 vs. SFB20: 0,82-0,99 (D5-D7)]. Para o EGF em cultivo, nenhuma diferença foi observada nos parâmetros avaliados. Em todos os experimentos, as curvas de crescimento foram típicas de forma S e as células passaram por uma fase lag, logarítmica e platô. Em conclusão, 20% de SFB é adequado para a recuperação de células somáticas; contudo, EGF não melhora a qualidade de crescimento dessas células. Ao nosso conhecimento, este é o primeiro estudo cultivando células somáticas de catetos.
45. Reduced representation bisulphite sequencing of ten bovine somatic tissues reveals DNA methylation patterns and their impacts on gene expression
- Author
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Leeson J. Alexander, El Hamidi abdel Hay, Tad S. Sonstegard, Derek M. Bickhart, Lingyang Xu, Curtis P. Van Tassell, Steven G. Schroeder, Hong Chen, Erin E. Connor, Yang Zhou, and George E. Liu
- Subjects
0301 basic medicine ,Epigenomics ,Bisulfite sequencing ,Gene Expression ,Biology ,Epigenesis, Genetic ,03 medical and health sciences ,chemistry.chemical_compound ,Cattle genome ,Genetics ,Animals ,Epigenetics ,DNA methylation ,Somatic tissue ,Methylation ,Sequence Analysis, DNA ,Molecular biology ,RRBS (reduced representation bisulphite sequencing) ,030104 developmental biology ,chemistry ,Organ Specificity ,Illumina Methylation Assay ,Cattle ,CpG Islands ,DNA microarray ,DNA ,Research Article ,Biotechnology - Abstract
Background As a major epigenetic component, DNA methylation plays important functions in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in economically important animals like cattle. Results Using reduced representation bisulphite sequencing (RRBS), we obtained single-base-resolution maps of bovine DNA methylation from ten somatic tissues. In total, we evaluated 1,868,049 cytosines in CG-enriched regions. While we found slightly low methylation levels (29.87 to 38.06 %) in cattle, the methylation contexts (CGs and non-CGs) of cattle showed similar methylation patterns to other species. Non-CG methylation was detected but methylation levels in somatic tissues were significantly lower than in pluripotent cells. To study the potential function of the methylation, we detected 10,794 differentially methylated cytosines (DMCs) and 836 differentially methylated CG islands (DMIs). Further analyses in the same tissues revealed many DMCs (including non-CGs) and DMIs, which were highly correlated with the expression of genes involved in tissue development. Conclusions In summary, our study provides a baseline dataset and essential information for DNA methylation profiles of cattle. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-3116-1) contains supplementary material, which is available to authorized users.
- Full Text
- View/download PDF
46. A Simple Cryopreservation Method for Efficient Isolation of Live Cells from Dead Animals
- Author
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Sano, Michiya, Kawanabe, Ayako, Kurosawa, Yaetsu, Suzuki, Yusuke, Takeda, Masahiro, Nakamura, Tomoaki, Iwata, Hisataka, Kuwayama, Takehito, and Shirasuna, Koumei
- Published
- 2022
- Full Text
- View/download PDF
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