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1.
Braz J Biol ; 83: e274888, 2023.
Article in English | MEDLINE | ID: mdl-38055577

ABSTRACT

Ateleia glazioveana Baill. is a pioneer, rustic and can be used for forest recovery. This work aimed to study the process of physiological maturation of this species. The research was carried out in the city of Alegre - ES, the trees were identified in the floral anthesis and accompanied during the filling of the fruits and development of the seeds until the complete maturation. The fruits were harvested at the following stages 7, 14, 21, 28, 35 and 42 days after anthesis, and characterized according to: morphometry, moisture, fresh and dry mass of fruits and seeds, germination, germination speed index, shoot and root length and dry mass of seedlings. The regression equations were adjusted for the main characteristics analyzed as a function of the harvest period. The point of physiological maturity of timbó occurred at 42 days after anthesis.


Subject(s)
Fabaceae , Seeds , Germination/physiology , Seedlings , Fruit
2.
DNA Repair (Amst) ; 129: 103532, 2023 09.
Article in English | MEDLINE | ID: mdl-37453244

ABSTRACT

Organisms have evolved a complex system, called the DNA damage response (DDR), which maintains genome integrity. The DDR is responsible for identifying and repairing a variety of lesions and alterations in DNA. DDR proteins coordinate DNA damage detection, cell cycle arrest, and repair, with many of these events regulated by protein phosphorylation. In the human proteome, 23 proteins contain the BRCT (BRCA1 C-Terminus domain) domain, a modular signaling domain that can bind phosphopeptides and mediate protein-protein interactions. BRCTs can be found as functional single units, tandem (tBRCT), triplet (tpBRCT), and quartet. Here we examine the evolution of the tpBRCT architecture present in TOPBP1 (DNA topoisomerase II binding protein 1) and ECT2 (epithelial cell transforming 2), and their respective interaction partners RAD9 (Cell cycle checkpoint control protein RAD9) and CYK-4 (Rac GTPase-activating protein 1), with a focus on the conservation of the phosphopeptide-binding residues. The pair TOPBP1-RAD9 arose with the Eukaryotes and ECT2-CYK-4 with the Eumetazoans. Triplet structural and functional characteristics were conserved in almost all organisms. The first unit of the triplet (BRCT0) is different from the other two BRCTs but conserved between orthologs for both TOPBP1 and ECT2. BRCT domain evolution simulations suggest a trend to retain the singlet or towards two or three BRCT copies per protein consistent with functional tBRCT and tpBRCT architectures. Our results shed light on the emergence of the function and architecture of multiple BRCT domain organizations and provide information about the evolution of the BRCT triplet. Knowledge of BRCT domain evolution can improve the understanding of DNA damage response mechanisms and signal transduction in DDR.


Subject(s)
BRCA1 Protein , Cell Cycle Proteins , Humans , BRCA1 Protein/metabolism , Protein Domains , Cell Cycle Proteins/metabolism , DNA Damage , Signal Transduction , Phosphorylation , Protein Binding
3.
Braz J Med Biol Res ; 55: e12381, 2022.
Article in English | MEDLINE | ID: mdl-36350974

ABSTRACT

Glial cells have been implicated in temporal lobe epilepsy in humans and in its models. Astrocytes are lost in several brain regions after acute seizures induced by pilocarpine and may suffer hyperplasia at subsequent time points. This study investigated the effect of N-methyl-(2S,4R)-trans-4-hydroxy-L-proline (NMP) on astrocytes exposed to cytotoxic concentrations of pilocarpine. Astrocytes were incubated with pilocarpine (half maximal inhibitory concentration (IC50)=31.86 mM) for 24 h. Afterwards, they were treated with NMP at concentrations ranging from 3.12 to 100 µg/mL for 24 h. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cytoplasmic reactive oxygen species (ROS) and mitochondrial transmembrane potential (ΔΨm) were analyzed by flow cytometry using 2',7'-dichlorofluorescein diacetate (DCFH-DA) and rhodamine-123 (Rho123), respectively. Expression of glial fibrillary acidic protein (GFAP) and voltage-dependent anion channel-1 (VDAC-1) were measured by western blot. Pilocarpine significantly decreased cell viability and mitochondrial potential and increased ROS concentration significantly by 6.7 times compared to the control. NMP concentrations ≥25 µg/mL protected astrocytes against pilocarpine-induced injury in a concentration-dependent manner. Concomitantly, NMP reduced cytoplasmic ROS accumulation to 27.3, 24.8, and 12.3% in the groups treated with 25, 50, and 100 µg/mL NMP, respectively. NMP also protected mitochondria from pilocarpine-induced depolarization. These effects were associated with improvement of pilocarpine-induced GFAP and VDAC-1 overexpression, which are important biomarkers of astrocyte dysfunction. In conclusion, the improvement of ROS accumulation, VDAC-1 overexpression, and mitochondrial depolarization are possible mechanisms of the NMP protective action on reactive astrocytes.


Subject(s)
Pilocarpine , Sapotaceae , Humans , Pilocarpine/pharmacology , Astrocytes , Reactive Oxygen Species/metabolism , Sapotaceae/metabolism
5.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;55: e12381, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1403908

ABSTRACT

Glial cells have been implicated in temporal lobe epilepsy in humans and in its models. Astrocytes are lost in several brain regions after acute seizures induced by pilocarpine and may suffer hyperplasia at subsequent time points. This study investigated the effect of N-methyl-(2S,4R)-trans-4-hydroxy-L-proline (NMP) on astrocytes exposed to cytotoxic concentrations of pilocarpine. Astrocytes were incubated with pilocarpine (half maximal inhibitory concentration (IC50)=31.86 mM) for 24 h. Afterwards, they were treated with NMP at concentrations ranging from 3.12 to 100 μg/mL for 24 h. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cytoplasmic reactive oxygen species (ROS) and mitochondrial transmembrane potential (ΔΨm) were analyzed by flow cytometry using 2',7'-dichlorofluorescein diacetate (DCFH-DA) and rhodamine-123 (Rho123), respectively. Expression of glial fibrillary acidic protein (GFAP) and voltage-dependent anion channel-1 (VDAC-1) were measured by western blot. Pilocarpine significantly decreased cell viability and mitochondrial potential and increased ROS concentration significantly by 6.7 times compared to the control. NMP concentrations ≥25 µg/mL protected astrocytes against pilocarpine-induced injury in a concentration-dependent manner. Concomitantly, NMP reduced cytoplasmic ROS accumulation to 27.3, 24.8, and 12.3% in the groups treated with 25, 50, and 100 µg/mL NMP, respectively. NMP also protected mitochondria from pilocarpine-induced depolarization. These effects were associated with improvement of pilocarpine-induced GFAP and VDAC-1 overexpression, which are important biomarkers of astrocyte dysfunction. In conclusion, the improvement of ROS accumulation, VDAC-1 overexpression, and mitochondrial depolarization are possible mechanisms of the NMP protective action on reactive astrocytes.

6.
Arq. bras. med. vet. zootec. (Online) ; 72(2): 443-451, Mar./Apr. 2020. ilus, tab
Article in Portuguese | VETINDEX | ID: vti-29635

ABSTRACT

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.(AU)


The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.(AU)


Subject(s)
Animals , Oocytes , Fertilization in Vitro/veterinary , In Vitro Oocyte Maturation Techniques/veterinary , Dasyproctidae , Parthenogenesis , Ionomycin
7.
Arq. bras. med. vet. zootec. (Online) ; 72(2): 443-451, Mar./Apr. 2020. ilus, tab
Article in Portuguese | LILACS, VETINDEX | ID: biblio-1128368

ABSTRACT

O objetivo foi avaliar protocolos de maturação in vitro (MIV) para oócitos de cutias, seguida de fertilização in vitro (FIV) e ativação partenogenética (AP). Os oócitos imaturos (CCOs) foram obtidos por fatiamento do ovário, após OSH, e submetidos a três grupos: MAT - 16 (16 horas de maturação), MAT - 20 (20 horas de maturação) e MAT - 24 (24 horas de maturação), em incubadora de cultivo a 38,8°C, com atmosfera de 5% de CO2 e 95% de umidade relativa. A maturação foi analisada pela presença do primeiro corpúsculo polar. Em seguida, os CCOs maduros foram submetidos à FIV, com período de coincubação dos CCOs e dos espermatozoides de 15h, a 38,8ºC e 5% de CO2, e AP com ionomicina. Os grupos de MIV foram analisados utilizando-se o teste qui-quadrado e, nos experimentos de FIV e AP, foram analisadas a taxa de clivagem e a proporção de desenvolvimento embrionário. A análise estatística foi realizada utilizando-se o programa SAS. Houve diferença significativa entre os grupos de maturação, tendo os grupos MAT - 20 e MAT - 24 apresentado maior porcentagem de oócitos maturados in vitro. As taxas de clivagem e de desenvolvimento embrionário foram de 8,6% e 2,9%, respectivamente, na FIV, e de 63,6% e 15,1%, na AP. Entretanto, nos dois casos, o embrião não passou do estágio de mórula.(AU)


The objective was to evaluate IVM protocols for agouti oocytes, followed by in vitro fertilization (IVF) and parthenogenetic activation (PA). The immature oocytes (CCOs) were obtained by slicing the ovary after OSH and submitted to three groups: MAT - 16 (16 hours maturation), MAT - 20 (20 hours maturation) and MAT - (24 hours maturation), in a culture incubator at 38.8°C, with an atmosphere of 5% CO2 and 95% relative humidity. The maturation was analyzed by the presence of the first polar corpuscle. Then, mature CCOs were submitted to IVF, with co-incubation period of CCOs and spermatozoa from 15h to 38.8°C and 5% of CO2, and PA with inomycin. The IVM groups were analyzed using the chi-square test and in the FIV and PA experiment the rate of cleavage and the rate of embryonic development were analyzed. Statistical analysis was performed using the SAS program. There was a significant difference between the maturation groups, and the MAT - 20 and MAT - 24 groups showed a higher percentage of matured oocytes in vitro. The rates of cleavage and embryonic development were 8.6% and 2.9%, respectively in FIV and 63.6% and 15.1% in PA. However, in both cases the embryo did not pass beyond the morula stage.(AU)


Subject(s)
Animals , Oocytes , Fertilization in Vitro/veterinary , In Vitro Oocyte Maturation Techniques/veterinary , Dasyproctidae , Parthenogenesis , Ionomycin
8.
Arq. bras. med. vet. zootec. (Online) ; 71(5): 1571-1581, set.-out. 2019. graf, ilus
Article in English | VETINDEX | ID: vti-25278

ABSTRACT

There is a growing interest in the study of unspecialized mesenchymal stem cells, for there are still some discussions about their in vitro behavior. Regenerative medicine is a science undergoing improvement which develops treatments as cell therapy using somatic stem cells. In several studies, adipose tissue is presented as a source of multipotent adult cells that has several advantages over other tissue sources. This study aimed to characterize and evaluate the tagging of mesenchymal stem cells from the agoutis adipose tissue (Dasyprocta prymonolopha), with fluorescent intracytoplasmic nanocrystals. Fibroblast cells were observed, plastic adherent, with extended self-renewal, ability to form colonies, multipotency by differentiation into three lineages, population CD90 + and CD45 - expression, which issued high red fluorescence after the tagging with fluorescent nanocrystals by different paths and cryopreserved for future use. It is possible to conclude that mesenchymal stem cells from agouti adipose tissue have biological characteristics and in vitro behavior that demonstrate its potential for use in clinical tests.(AU)


Há um interesse crescente no estudo das células estaminais mesenquimais, não especializadas, pois ainda existem algumas discussões sobre seu comportamento in vitro. A medicina regenerativa é uma ciência em fase de crescimento que desenvolve tratamentos como terapia celular utilizando células estaminais somáticas. Em vários estudos, o tecido adiposo é apresentado como uma fonte de células adultas multipotentes que tem várias vantagens em relação a outras fontes de tecido. Este estudo teve como objetivo caracterizar e avaliar a marcação de células estaminais mesenquimais do tecido adiposo de cutias (Dasyprocta prymnolopha) com nanocristais intracitoplasmáticos fluorescentes. Observaram-se células fibroblásticas, aderentes ao plástico, com autorrenovação prolongada, capacidade de formar colônias, diferenciação em três linhagens, população CD90 + e expressão CD45, que emitiram alta fluorescência vermelha após a marcação com nanocristais fluorescentes por diferentes vias, e criopreservadas para uso futuro. É possível concluir que as células estaminais mesenquimais do tecido adiposo de cutias têm características biológicas e comportamentos in vitro que demonstram seu potencial para uso em testes clínicos.(AU)


Subject(s)
Animals , Adipose Tissue/cytology , Immunophenotyping/veterinary , Regenerative Medicine/methods , Nanoparticles , Mesenchymal Stem Cells , Dasyproctidae/genetics
9.
Arq. bras. med. vet. zootec. (Online) ; 71(5): 1571-1581, set.-out. 2019. graf, ilus
Article in English | VETINDEX, LILACS | ID: biblio-1038673

ABSTRACT

There is a growing interest in the study of unspecialized mesenchymal stem cells, for there are still some discussions about their in vitro behavior. Regenerative medicine is a science undergoing improvement which develops treatments as cell therapy using somatic stem cells. In several studies, adipose tissue is presented as a source of multipotent adult cells that has several advantages over other tissue sources. This study aimed to characterize and evaluate the tagging of mesenchymal stem cells from the agoutis adipose tissue (Dasyprocta prymonolopha), with fluorescent intracytoplasmic nanocrystals. Fibroblast cells were observed, plastic adherent, with extended self-renewal, ability to form colonies, multipotency by differentiation into three lineages, population CD90 + and CD45 - expression, which issued high red fluorescence after the tagging with fluorescent nanocrystals by different paths and cryopreserved for future use. It is possible to conclude that mesenchymal stem cells from agouti adipose tissue have biological characteristics and in vitro behavior that demonstrate its potential for use in clinical tests.(AU)


Há um interesse crescente no estudo das células estaminais mesenquimais, não especializadas, pois ainda existem algumas discussões sobre seu comportamento in vitro. A medicina regenerativa é uma ciência em fase de crescimento que desenvolve tratamentos como terapia celular utilizando células estaminais somáticas. Em vários estudos, o tecido adiposo é apresentado como uma fonte de células adultas multipotentes que tem várias vantagens em relação a outras fontes de tecido. Este estudo teve como objetivo caracterizar e avaliar a marcação de células estaminais mesenquimais do tecido adiposo de cutias (Dasyprocta prymnolopha) com nanocristais intracitoplasmáticos fluorescentes. Observaram-se células fibroblásticas, aderentes ao plástico, com autorrenovação prolongada, capacidade de formar colônias, diferenciação em três linhagens, população CD90 + e expressão CD45, que emitiram alta fluorescência vermelha após a marcação com nanocristais fluorescentes por diferentes vias, e criopreservadas para uso futuro. É possível concluir que as células estaminais mesenquimais do tecido adiposo de cutias têm características biológicas e comportamentos in vitro que demonstram seu potencial para uso em testes clínicos.(AU)


Subject(s)
Animals , Adipose Tissue/cytology , Immunophenotyping/veterinary , Regenerative Medicine/methods , Nanoparticles , Mesenchymal Stem Cells , Dasyproctidae/genetics
10.
Theriogenology ; 125: 56-63, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30388472

ABSTRACT

The aim of this study was to evaluate the effect of the conditioned medium of ovine Wharton's jelly-derived mesenchymal stem cells (oWJ-MSCs) on the morphology, growth, reactive oxygen species (ROS) and glutathione (GSH) intracellular levels, active mitochondria, and meiotic resumption of isolated ovine secondary follicles in vitro. The oWJ-MSCs were isolated and the medium where they were cultured was recovered (conditioned medium). Isolated ovine secondary follicles were cultured for 6 days in 1) supplemented α-MEM+ (control); 2) 50% α-MEM+ + 50% conditioned medium (α-MEM + CM group) or 3) conditioned medium only (CM group). The parameters analyzed were morphology, antrum formation, follicle and oocyte growth, ROS and GSH levels, mitochondrial activity and meiotic resumption. The percentage of normal follicles, antrum formation, and fully grown oocytes did not differ (P > 0.05) among treatments. Follicles cultured in α-MEM + CM group had greater (P < 0.05) diameter than other treatments after culture. Moreover, the diameter of the follicles cultured in CM alone was higher (P < 0.05) than in the α-MEM+. In addition, α-MEM + CM and CM treatments increased the growth rate compared to the α-MEM+. Treatments containing conditioned medium (α-MEM + CM or CM) significantly reduced ROS levels compared to the control medium. Moreover, mitochondrial activity was higher in α-MEM+ and α-MEM + CM than in CM alone. All treatments showed oocytes in GV, GVBD and MI. In conclusion, oWJ-MSCs conditioned medium, especially when associated with α-MEM, improves the growth of secondary follicles and reduces ROS generation after short-term culture.


Subject(s)
Culture Media, Conditioned , Mesenchymal Stem Cells/physiology , Ovarian Follicle/physiology , Reactive Oxygen Species/metabolism , Sheep/physiology , Wharton Jelly/cytology , Animals , Female , Tissue Culture Techniques
11.
R. bras. Ci. avíc. ; 21(1): eRBCA-2019-0687, abr. 2019. tab, graf
Article in English | VETINDEX | ID: vti-17600

ABSTRACT

The objective of this study was to determine if lipid extraction processes alter the isotopic value of 13C and 15N of tissues (pectoral muscle, thigh and liver) and eggs and if the use of anticoagulants interferes with blood and plasma 13C and 15N isotopic values. Samples were acquired from the same flock of birds. The 32 egg samples were randomly divided into four treatments (liquid, dehydrated, and fat-extracted with ether or chloroform + methanol) with eight replicates each. The 24 samples of pectoral muscle, thigh muscle and liver of broilers were randomly divided into three treatments (dehydrated, fat-extracted with ether and chloroform + methanol) with eight replicates each. Blood samples were divided into a 3x3 factorial arrangement with three physical forms (liquid, oven-dried or freeze-dried) and three collection methods (with no anticoagulant, with EDTA or heparin). Plasma samples were distributed in a 3x2 factorial arrangement, with three physical forms (liquid, oven-dried, or freeze-dried) and two anticoagulants (EDTA or heparin). The obtained isotopic results were submitted to the multivariate analysis of variance (MANOVA) and univariate (ANOVA, complemented by Tukey test), using the GLM procedure of the statistical program SAS (1996) or Minitab 16. The results show that it is possible to use the evaluated methods of fat extraction, drying and anticoagulants in the isotopic analyses of carbon-13 and nitrogen-15 in chicken tissues.(AU)


Subject(s)
Animals , Birds , Isotope Labeling/methods , Isotope Labeling/veterinary , Carbon
12.
Rev. bras. ciênc. avic ; 21(1): eRBCA, abr. 2019. tab, graf
Article in English | VETINDEX | ID: biblio-1490602

ABSTRACT

The objective of this study was to determine if lipid extraction processes alter the isotopic value of 13C and 15N of tissues (pectoral muscle, thigh and liver) and eggs and if the use of anticoagulants interferes with blood and plasma 13C and 15N isotopic values. Samples were acquired from the same flock of birds. The 32 egg samples were randomly divided into four treatments (liquid, dehydrated, and fat-extracted with ether or chloroform + methanol) with eight replicates each. The 24 samples of pectoral muscle, thigh muscle and liver of broilers were randomly divided into three treatments (dehydrated, fat-extracted with ether and chloroform + methanol) with eight replicates each. Blood samples were divided into a 3x3 factorial arrangement with three physical forms (liquid, oven-dried or freeze-dried) and three collection methods (with no anticoagulant, with EDTA or heparin). Plasma samples were distributed in a 3x2 factorial arrangement, with three physical forms (liquid, oven-dried, or freeze-dried) and two anticoagulants (EDTA or heparin). The obtained isotopic results were submitted to the multivariate analysis of variance (MANOVA) and univariate (ANOVA, complemented by Tukey test), using the GLM procedure of the statistical program SAS (1996) or Minitab 16. The results show that it is possible to use the evaluated methods of fat extraction, drying and anticoagulants in the isotopic analyses of carbon-13 and nitrogen-15 in chicken tissues.


Subject(s)
Animals , Birds , Isotope Labeling/methods , Isotope Labeling/veterinary , Carbon
13.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;51(2): e6546, 2018. graf
Article in English | LILACS | ID: biblio-889018

ABSTRACT

The inducible inflammatory enzyme cycloxigenase-2 is up-regulated in cancer, and favors tumor progression. Cycloxigenase-2 is encoded by the prostaglandin-endoperoxide synthase 2 (PTGS2) gene, which presents sequence variations in the promoter region (PR) and in the 3′-untranslated region (3′-UTR). Different PR (rs689465, rs689466, rs20417) and 3′-UTR (rs5275) variants were generated by site-directed mutagenesis, and combined in haplotypes to access expression levels using a reporter system (luciferase) in human cells (MCF-7 and HEK293FT). Luciferase activity did not differ significantly among PTGS2 PR constructs, except for pAAC (containing variant allele rs20417 C), with 40% less activity than pAAG (wild-type sequence) in MCF-7 cells (P<0.01). Despite the lack of individual significant differences, PTGS2 PR constructs enclosing rs689466 G (pAGG and pAGC) showed an approximate two-fold increase in luciferase activity when compared to those containing rs689466 A (pAAG, pGAC, pAAC and pGAG) in both cell lines (P<0.001 for MCF-7 and P=0.03 for HEK293FT). The effect of PTGS2 3′-UTR sequences varied between MCF-7 and HEK293FT: MCF-7 cells showed significant reduction (40-60%) in luciferase activity (at least P<0.01), whereas HEK293FT cells showed more diverse results, with an average 2-fold increase when combined constructs (PR and 3′-UTR) were compared to respective parental PR sequences. The contribution of 3′-UTR variant (rs5275) was not consistent in either cell line. Despite the modulation of the 3′-UTR, with variable effects of rs5275, the enhancing transcriptional effect of rs689466 G was still detectable (P<0.0001 in MCF-7 or P=0.03 in HEK293FT cells).


Subject(s)
Humans , Gene Expression Regulation, Neoplastic/genetics , Cyclooxygenase 2/genetics , Haplotypes , Up-Regulation , Mutagenesis, Site-Directed , Polymorphism, Single Nucleotide , Cell Line, Tumor , Cyclooxygenase 2/metabolism , MCF-7 Cells , Genotype , Luciferases/metabolism
14.
Braz J Med Biol Res ; 51(2): e6546, 2017 Nov 30.
Article in English | MEDLINE | ID: mdl-29211250

ABSTRACT

The inducible inflammatory enzyme cycloxigenase-2 is up-regulated in cancer, and favors tumor progression. Cycloxigenase-2 is encoded by the prostaglandin-endoperoxide synthase 2 (PTGS2) gene, which presents sequence variations in the promoter region (PR) and in the 3'-untranslated region (3'-UTR). Different PR (rs689465, rs689466, rs20417) and 3'-UTR (rs5275) variants were generated by site-directed mutagenesis, and combined in haplotypes to access expression levels using a reporter system (luciferase) in human cells (MCF-7 and HEK293FT). Luciferase activity did not differ significantly among PTGS2 PR constructs, except for pAAC (containing variant allele rs20417 C), with 40% less activity than pAAG (wild-type sequence) in MCF-7 cells (P<0.01). Despite the lack of individual significant differences, PTGS2 PR constructs enclosing rs689466 G (pAGG and pAGC) showed an approximate two-fold increase in luciferase activity when compared to those containing rs689466 A (pAAG, pGAC, pAAC and pGAG) in both cell lines (P<0.001 for MCF-7 and P=0.03 for HEK293FT). The effect of PTGS2 3'-UTR sequences varied between MCF-7 and HEK293FT: MCF-7 cells showed significant reduction (40-60%) in luciferase activity (at least P<0.01), whereas HEK293FT cells showed more diverse results, with an average 2-fold increase when combined constructs (PR and 3'-UTR) were compared to respective parental PR sequences. The contribution of 3'-UTR variant (rs5275) was not consistent in either cell line. Despite the modulation of the 3'-UTR, with variable effects of rs5275, the enhancing transcriptional effect of rs689466 G was still detectable (P<0.0001 in MCF-7 or P=0.03 in HEK293FT cells).


Subject(s)
3' Untranslated Regions/genetics , Cyclooxygenase 2/genetics , Gene Expression Regulation, Neoplastic/genetics , Cell Line, Tumor , Cyclooxygenase 2/metabolism , Genotype , Haplotypes , Humans , Luciferases/metabolism , MCF-7 Cells , Mutagenesis, Site-Directed , Polymorphism, Single Nucleotide , Up-Regulation
15.
Braz. J. Biol. ; 77(4): 868-875, Nov. 2017. tab, ilus
Article in English | VETINDEX | ID: vti-20371

ABSTRACT

Roots and leaves of Panicum maximum Tanzânia, Mombaça and Massai; Urochloa brizantha Piatã, Marandu and Xaraés; Urochloa humidicola Llanero; Urochloa ruziziensis Ruzizienses; Urochloa hybrida Mulato II and Cynodon nlemfuensis Estrela-roxa were analyzed, seeking to identify characters for better adaptation to the environment that may interfere with digestibility of tissue from the point of view of the rumen in cattle. Were planted ten cultivars in a completely randomized blocks with three repetitions. Was collected vegetative material, which histological slides were prepared from middle third of the sections of roots and leaves. Were observed differences (p>0.05) in the roots: higher volume of epidermal cells (28.62 µm) and overall diameter (1926.41 µm) of Llanero; thicker vascular cylinder (975.09 µm) and more protoxylem (42.25) in Estrela-roxa and occurrence of aerenchyma in cultivars Piatã, Mulato II, Xaraés, Massai, Llanero and Estrela-roxa; Were found higher proportions of bulliform cells in the leaves (121.07 µm) and thicker leaf mesophyll in U. humidicola Llanero (263.63 µm); higher proportion of sclerenchyma fibers in Xaraés and Marandu; lower results for amount of fibers in P. maximum Massai. We conclude that the cultivars Estrela-roxa, Llanero and Massai have greater adaptability to the environment and better nutritional quality.(AU)


Foram analisadas raízes e folhas de Panicum maximum Tanzânia, Mombaça e Massai; Urochloa brizantha Piatã, Marandu e Xaraés; Urochloa humidicola Llanero; Urochloa ruziziensis Ruzizienses; Urochloa hybrida Mulato II e Cynodon nlemfuensis Estrela-roxa, procurando identificar caracteres relacionados à melhor adaptação ao ambiente e à qualidade nutritiva das forrageiras. As dez cultivares foram semeadas em blocos inteiramente casualizados com três repetições. Após estabelecidas foi coletado material vegetativo, do qual lâminas histológicas foram confeccionadas a partir de secções do terço médio de raízes e folhas. Foram observadas diferenças significativas (p>0,05) nas raízes: maior volume de células epidérmicas (28,62 µm) e diâmetro total (1926,41 µm) de Llanero; em Estrela-roxa maior espessura do cilindro vascular (975,09 µm) e número maior de protoxilemas (42,25) e formação de aerênquimas nas cultivares Piatã, Mulato II, Xaraés, Massai, Llanero, e Estrela-roxa; Nas folhas, foram constatadas maiores proporções de células buliformes (121,07 µm) e mesofilo foliar mais espesso (263,63 µm) em Llanero; em Xaraés e Marandu maiores proporções de fibras esclerenquimáticas; em Massai menores resultados para quantidade de fibras. Conclui-se que as cultivares Estrela-roxa, Llanero e Massai apresentam maior adaptabilidade ao ambiente e melhor qualidade nutritiva.(AU)

16.
Braz. j. biol ; Braz. j. biol;77(4): 868-875, Nov. 2017. tab, graf
Article in English | LILACS | ID: biblio-888816

ABSTRACT

Abstract Roots and leaves of Panicum maximum Tanzânia, Mombaça and Massai; Urochloa brizantha Piatã, Marandu and Xaraés; Urochloa humidicola Llanero; Urochloa ruziziensis Ruzizienses; Urochloa hybrida Mulato II and Cynodon nlemfuensis Estrela-roxa were analyzed, seeking to identify characters for better adaptation to the environment that may interfere with digestibility of tissue from the point of view of the rumen in cattle. Were planted ten cultivars in a completely randomized blocks with three repetitions. Was collected vegetative material, which histological slides were prepared from middle third of the sections of roots and leaves. Were observed differences (p>0.05) in the roots: higher volume of epidermal cells (28.62 µm) and overall diameter (1926.41 µm) of Llanero; thicker vascular cylinder (975.09 µm) and more protoxylem (42.25) in Estrela-roxa and occurrence of aerenchyma in cultivars Piatã, Mulato II, Xaraés, Massai, Llanero and Estrela-roxa; Were found higher proportions of bulliform cells in the leaves (121.07 µm) and thicker leaf mesophyll in U. humidicola Llanero (263.63 µm); higher proportion of sclerenchyma fibers in Xaraés and Marandu; lower results for amount of fibers in P. maximum Massai. We conclude that the cultivars Estrela-roxa, Llanero and Massai have greater adaptability to the environment and better nutritional quality.


Resumo Foram analisadas raízes e folhas de Panicum maximum Tanzânia, Mombaça e Massai; Urochloa brizantha Piatã, Marandu e Xaraés; Urochloa humidicola Llanero; Urochloa ruziziensis Ruzizienses; Urochloa hybrida Mulato II e Cynodon nlemfuensis Estrela-roxa, procurando identificar caracteres relacionados à melhor adaptação ao ambiente e à qualidade nutritiva das forrageiras. As dez cultivares foram semeadas em blocos inteiramente casualizados com três repetições. Após estabelecidas foi coletado material vegetativo, do qual lâminas histológicas foram confeccionadas a partir de secções do terço médio de raízes e folhas. Foram observadas diferenças significativas (p>0,05) nas raízes: maior volume de células epidérmicas (28,62 µm) e diâmetro total (1926,41 µm) de Llanero; em Estrela-roxa maior espessura do cilindro vascular (975,09 µm) e número maior de protoxilemas (42,25) e formação de aerênquimas nas cultivares Piatã, Mulato II, Xaraés, Massai, Llanero, e Estrela-roxa; Nas folhas, foram constatadas maiores proporções de células buliformes (121,07 µm) e mesofilo foliar mais espesso (263,63 µm) em Llanero; em Xaraés e Marandu maiores proporções de fibras esclerenquimáticas; em Massai menores resultados para quantidade de fibras. Conclui-se que as cultivares Estrela-roxa, Llanero e Massai apresentam maior adaptabilidade ao ambiente e melhor qualidade nutritiva.


Subject(s)
Animals , Poaceae/anatomy & histology , Animal Husbandry , Brazil , Cattle , Plant Roots/anatomy & histology , Plant Leaves/anatomy & histology , Acclimatization
17.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 997-1005, jul.-ago. 2017. ilus, tab, graf
Article in Portuguese | VETINDEX | ID: vti-18021

ABSTRACT

Este trabalho objetivou apresentar a caracterização da morfologia do testículo de cutia (Dasyproctaprymnolopha) macho, com o intuito de colaborar com o conhecimento da morfofisiologia reprodutiva da espécie. Foram utilizados testículos de 47 animais, com idade entre um e dois anos, pesos homogêneos (2,08 ± 0,23kg), oriundos do Núcleo de Estudos e Preservação de Animais Silvestres do Centro de Ciências Agrárias da Universidade Federal do Piauí. As estruturas foram dissecadas, descritas, e fragmentos foram processados para a microscopia de luz, sendo, posteriormente avaliada a atividade gonadal. Observou-se que os testículos são órgãos elipsoides alongados, podendo ser encontrados na região inguinal ou na cavidade abdominal, não apresentando um escroto bem delimitado. Verificou-se também parênquima com característica histológica padrão para o órgão em mamíferos, com a identificação de oito tipos de associações celulares, caracterizando os estádios do ciclo do epitélio seminífero, com menor e maior frequência dos estádios 3 e 5, respectivamente.(AU)


This study meant to characterize the morphology of the testicle from (Dasyprocta prymnolopha) agouti males, in order to collaborate with the knowledge of reproductive morphophysiology of the specie. Testicles were used from 47 animals aged between 1 and 2 years, homogeneous weight (2.08±0.23kg), coming from the Centre for the Study and Conservation of Wild Animals of Agricultural Sciences Center of the Federal University of Piauí. The structures were dissected, described and fragments were processed for light microscopy, and, subsequently, gonadal activity was evaluated. Testes were observed to be elongated ellipsoidal bodies that can be found in the groin or in the abdominal cavity, not having a clearly defined scrotum. We also could see parenchymal with standard histological characteristic for the mammalian body, with the identification of eight types of cell associations, characterized epithelium Seminiferous stages of the cycle, with lower and higher frequency of stages 3 and 5, respectively.(AU)


Subject(s)
Animals , Male , Dasyproctidae/anatomy & histology , Testis/anatomy & histology , Spermatogenesis , Body Weights and Measures/veterinary
18.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 980-988, jul.-ago. 2017. tab
Article in Portuguese | VETINDEX | ID: vti-18019

ABSTRACT

O objetivo da presente pesquisa foi alcançado com a divisão da pesquisa em dois experimentos: (1) aperfeiçoar o protocolo de congelação utilizando água de coco em pó (ACP-104) como diluente para a criopreservação seminal de carpa comum; (2) avaliar o efeito da suplementação das vitaminas C (ácido ascórbico) ou E (α-tocoferol) sobre os melhores diluidores testados no experimento 1 na qualidade do sêmen pós-descongelado da espécie. Para o experimento 1, foram formados oito pools de sêmen, provenientes de 14 machos selecionados. As amostras seminais coletadas foram avaliadas quanto à motilidade total, à velocidade, ao percentual de espermatozoides normais e à vitalidade espermática antes e depois da criopreservação seminal. Esta foi realizada em meio ACP-104 acrescido de dimetilsulfóxido (DMSO), ou etilenoglicol (EG), ou glicerol, ou metanol, todos à concentração de 10%, diluídos em 1:3 (sêmen:diluidor). As amostras foram, então, congeladas em vapor de nitrogênio líquido em dry shipper e estocadas em nitrogênio líquido (-196°C). Para o experimento 2, foram formados oito pools provenientes da coleta de sêmen de 15 machos. As amostras seminais foram avaliadas seguindo as mesmas análises do experimento 1, acrescentando-se a duração da motilidade total. A criopreservação seminal utilizou-se do meio ACP-104 acrescido de DMSO ou EG, suplementado ou não com vitamina C ou E. Os melhores resultados encontrados no experimento 1 foram obtidos com o DMSO e o EG. Estes não diferiram significativamente entre si para a motilidade total (24% e 28%; P>0,05) e a normalidade espermática (32% e 26%; P>0,05), respectivamente. Para o experimento 2, o EG suplementado com vitamina E produziu significativamente resultados superiores de motilidade total, normalidade espermática e duração da motilidade em relação ao DMSO, concluindo-se que o EG deve ser, portanto, o crioprotetor de escolha a ser utilizado com o ACP-104 suplementado ou não com vitamina E.(AU)


The objective was achieved by dividing the research into two experiments: (1) improving the freezing protocol using powdered coconut water (ACP-104) as a diluent for the cryopreservation seminal of common carp; (2) evaluating the effect of supplementation of vitamins C (ascorbic acid) or vitamin E (α-tocoferol) with the best extenders tested in experiment 1 on the quality of post-thawed. For experiment 1, semen pools from 14 selected males were formed. Seminal samples were evaluated for total motility, velocity, percentage of normal sperm and sperm vitality before and after the seminal cryopreservation. This was done in ACP-104 extender plus dimethyl sulfoxide (DMSO), or ethylene glycol (EG), or glycerol or methanol all at concentration 10% diluted in 1:3 (semen:extender). The samples were frozen in vapors of nitrogen into dry shippers and stored in liquid nitrogen (-196 °C). For experiment 2, eight pools were formed from the 15 males. The semen samples were evaluated following the same analysis of experiment 1 adding duration of total motility. The sperm cryopreservation was performed in extenders ACP-104 plus DMSO or EG supplemented or not with vitamin C or E. The best results found in Experiment 1 were obtained with DMSO and EG. They do not differ significantly for total motility (24% and 28%; P>0.05) and normal sperm (32% and 26%; P>0.05) respectively. For experiment 2, EG supplemented with vitamin E, produced significantly better results overall motility, sperm normality and duration of motility relative to DMSO. In conclusion, EG should be the cryoprotectant of choice for use with the ACP-104 supplemented or not with vitamin E.(AU)


Subject(s)
Animals , Vitamins/administration & dosage , Cryopreservation/veterinary , Semen Preservation/veterinary , Semen Analysis/veterinary , Antioxidants/analysis , Carps
19.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 980-988, jul.-ago. 2017. tab
Article in Portuguese | LILACS, VETINDEX | ID: biblio-876722

ABSTRACT

O objetivo da presente pesquisa foi alcançado com a divisão da pesquisa em dois experimentos: (1) aperfeiçoar o protocolo de congelação utilizando água de coco em pó (ACP-104) como diluente para a criopreservação seminal de carpa comum; (2) avaliar o efeito da suplementação das vitaminas C (ácido ascórbico) ou E (α-tocoferol) sobre os melhores diluidores testados no experimento 1 na qualidade do sêmen pós-descongelado da espécie. Para o experimento 1, foram formados oito pools de sêmen, provenientes de 14 machos selecionados. As amostras seminais coletadas foram avaliadas quanto à motilidade total, à velocidade, ao percentual de espermatozoides normais e à vitalidade espermática antes e depois da criopreservação seminal. Esta foi realizada em meio ACP-104 acrescido de dimetilsulfóxido (DMSO), ou etilenoglicol (EG), ou glicerol, ou metanol, todos à concentração de 10%, diluídos em 1:3 (sêmen:diluidor). As amostras foram, então, congeladas em vapor de nitrogênio líquido em dry shipper e estocadas em nitrogênio líquido (-196°C). Para o experimento 2, foram formados oito pools provenientes da coleta de sêmen de 15 machos. As amostras seminais foram avaliadas seguindo as mesmas análises do experimento 1, acrescentando-se a duração da motilidade total. A criopreservação seminal utilizou-se do meio ACP-104 acrescido de DMSO ou EG, suplementado ou não com vitamina C ou E. Os melhores resultados encontrados no experimento 1 foram obtidos com o DMSO e o EG. Estes não diferiram significativamente entre si para a motilidade total (24% e 28%; P>0,05) e a normalidade espermática (32% e 26%; P>0,05), respectivamente. Para o experimento 2, o EG suplementado com vitamina E produziu significativamente resultados superiores de motilidade total, normalidade espermática e duração da motilidade em relação ao DMSO, concluindo-se que o EG deve ser, portanto, o crioprotetor de escolha a ser utilizado com o ACP-104 suplementado ou não com vitamina E.(AU)


The objective was achieved by dividing the research into two experiments: (1) improving the freezing protocol using powdered coconut water (ACP-104) as a diluent for the cryopreservation seminal of common carp; (2) evaluating the effect of supplementation of vitamins C (ascorbic acid) or vitamin E (α-tocoferol) with the best extenders tested in experiment 1 on the quality of post-thawed. For experiment 1, semen pools from 14 selected males were formed. Seminal samples were evaluated for total motility, velocity, percentage of normal sperm and sperm vitality before and after the seminal cryopreservation. This was done in ACP-104 extender plus dimethyl sulfoxide (DMSO), or ethylene glycol (EG), or glycerol or methanol all at concentration 10% diluted in 1:3 (semen:extender). The samples were frozen in vapors of nitrogen into dry shippers and stored in liquid nitrogen (-196 °C). For experiment 2, eight pools were formed from the 15 males. The semen samples were evaluated following the same analysis of experiment 1 adding duration of total motility. The sperm cryopreservation was performed in extenders ACP-104 plus DMSO or EG supplemented or not with vitamin C or E. The best results found in Experiment 1 were obtained with DMSO and EG. They do not differ significantly for total motility (24% and 28%; P>0.05) and normal sperm (32% and 26%; P>0.05) respectively. For experiment 2, EG supplemented with vitamin E, produced significantly better results overall motility, sperm normality and duration of motility relative to DMSO. In conclusion, EG should be the cryoprotectant of choice for use with the ACP-104 supplemented or not with vitamin E.(AU)


Subject(s)
Animals , Antioxidants/analysis , Carps , Cryopreservation/veterinary , Semen Analysis/veterinary , Semen Preservation/veterinary , Vitamins/administration & dosage
20.
Arq. bras. med. vet. zootec. (Online) ; 69(4): 997-1005, jul.-ago. 2017. ilus, tab, graf
Article in Portuguese | LILACS, VETINDEX | ID: biblio-876727

ABSTRACT

Este trabalho objetivou apresentar a caracterização da morfologia do testículo de cutia (Dasyproctaprymnolopha) macho, com o intuito de colaborar com o conhecimento da morfofisiologia reprodutiva da espécie. Foram utilizados testículos de 47 animais, com idade entre um e dois anos, pesos homogêneos (2,08 ± 0,23kg), oriundos do Núcleo de Estudos e Preservação de Animais Silvestres do Centro de Ciências Agrárias da Universidade Federal do Piauí. As estruturas foram dissecadas, descritas, e fragmentos foram processados para a microscopia de luz, sendo, posteriormente avaliada a atividade gonadal. Observou-se que os testículos são órgãos elipsoides alongados, podendo ser encontrados na região inguinal ou na cavidade abdominal, não apresentando um escroto bem delimitado. Verificou-se também parênquima com característica histológica padrão para o órgão em mamíferos, com a identificação de oito tipos de associações celulares, caracterizando os estádios do ciclo do epitélio seminífero, com menor e maior frequência dos estádios 3 e 5, respectivamente.(AU)


This study meant to characterize the morphology of the testicle from (Dasyprocta prymnolopha) agouti males, in order to collaborate with the knowledge of reproductive morphophysiology of the specie. Testicles were used from 47 animals aged between 1 and 2 years, homogeneous weight (2.08±0.23kg), coming from the Centre for the Study and Conservation of Wild Animals of Agricultural Sciences Center of the Federal University of Piauí. The structures were dissected, described and fragments were processed for light microscopy, and, subsequently, gonadal activity was evaluated. Testes were observed to be elongated ellipsoidal bodies that can be found in the groin or in the abdominal cavity, not having a clearly defined scrotum. We also could see parenchymal with standard histological characteristic for the mammalian body, with the identification of eight types of cell associations, characterized epithelium Seminiferous stages of the cycle, with lower and higher frequency of stages 3 and 5, respectively.(AU)


Subject(s)
Animals , Male , Body Weights and Measures/veterinary , Dasyproctidae/anatomy & histology , Spermatogenesis , Testis/anatomy & histology
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