RESUMO
Cereus hildmannianus is a cactus exhibiting morphological and physiological adaptation of its cladodes which ensuring growth in climatic and soil conditions unfavourable for many plant species. Currently, limited water resources and increasing demand for renewable energy make cacti a biomass source for the production of biofuels. Somaclones regenerated from callus in vitro can be a source of new raw material in useful plants. The objective of this work was to determine if the regenerated plants showing two different morphologies present polysaccharide composition different from the wild plant. Somaclones aqueous extraction shows the absence of soluble polysaccharides as mucilage. The alkaline extraction of in vivo cultivated plant showed the presence of starch, type I arabinogalactan, and arabinoxylan and the somaclones showed type I arabinogalactan and arabinoxylan in both morphologies. Hemicelluloses found in the somaclones are not different from in vivo cultivated plants, but somaclones not almost biosynthesize mucilage and starch.
Assuntos
Cactaceae , Cactaceae/metabolismo , Parede Celular/metabolismo , Plantas/metabolismo , Polissacarídeos , AmidoRESUMO
Protease-activated receptors (PARs) are G protein-coupled receptors, which are activated by proteolytical cleavage of the amino-terminus and act as sensors for extracellular proteases. We hypothesized that PAR-1 and PAR-2 can be modulated by inflammatory stimulus in human dental pulp cells. PAR-1 and PAR-2 gene expression in human pulp tissue and MDPC-23 cells were analyzed by quantitative polymerase chain reaction. Monoclonal PAR-1 and PAR-2 antibodies were used to investigate the cellular expression of these receptors using Western blot, flow cytometry, and confocal microscopy in MDPC-23 cells. Immunofluorescence assays of human intact and carious teeth were performed to assess the presence of PAR-1 and PAR-2 in the dentin-pulp complex. The results show for the first time that human odontoblasts and MDPC-23 cells constitutively express PAR-1 and PAR-2. PAR-2 activation increased significantly the messenger RNA expression of matrix metalloproteinase (MMP)-2, MMP-9, MMP-13, and MMP-14 in MDPC-23 cells ( P < 0.05), while the expression of these enzymes decreased significantly in the PAR-1 agonist group ( P < 0.05). The high-performance liquid chromatography and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis showed the presence of MMP-13 activity cleaving PAR-1 at specific, noncanonical site TLDPRS42↓F43LL in human dental pulp tissues. Also, we detected a presence of a trypsin-like activity cleaving PAR-2 at canonical site SKGR20↓S21LIGRL in pulp tissues. Confocal microscopy analysis of human dentin-pulp complex showed intense positive staining of PAR-1 and PAR-2 in the odontoblast processes in dentinal tubules of carious teeth compared to intact ones. The present results support the hypothesis of activation of the upregulated PAR-1 and PAR-2 by endogenous proteases abundant during the inflammatory response in dentin-pulp complex.
Assuntos
Polpa Dentária/citologia , Odontoblastos/enzimologia , Receptor PAR-1/metabolismo , Receptor PAR-2/metabolismo , Adulto , Western Blotting , Cromatografia Líquida de Alta Pressão , Citometria de Fluxo , Humanos , Inflamação/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Microscopia Confocal , RNA Mensageiro/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Regulação para CimaRESUMO
Knowledge of genetic diversity among genotypes and relationships among elite lines is of great importance for the development of breeding programs. Therefore, the objective of this study was to evaluate genetic variability based on the morphoagronomic and molecular characterization of 18 elite popcorn (Zea mays var. everta) lines to be used by Universidade Estadual de Maringá breeding programs. We used 31 microsatellite primers (widely distributed in the genome), and 16 morphological descriptors (including the resistance to maize white spot, common rust, polysora rust of maize, cercospora and leaf blights). The molecular data revealed variability among the lines, which were divided into four groups that were partially concordant with unweighted pair group method with arithmetic mean (UPMGA) and Bayesian clusters. The lines G3, G4, G11, and G13 exhibited favorable morphological characters and low disease incidence rates. The four groups were confirmed using the Gower distance in the UPGMA cluster; however, there was no association with the dissimilarity patterns obtained using the molecular data. The absence of a correlation suggests that both characterizations (morphoagronomic and molecular) are important for discriminating among elite popcorn lines.
Assuntos
Polimorfismo Genético , Zea mays/genética , Fungos/patogenicidade , Repetições de Microssatélites , Melhoramento Vegetal , Imunidade Vegetal/imunologia , Característica Quantitativa Herdável , Zea mays/crescimento & desenvolvimento , Zea mays/imunologiaRESUMO
The reduction in sugarcane productivity in subsequent cutting stages may be related to a gradual decrease of the allele number and mean observed heterozygosity (HO) in the sugarcane ratoon. This hypothesis was tested assessing the number of alleles and HO values in 10 expressed sequence tag microsatellites (Est-SSR loci) of the sugarcane varieties RB72454 and RB867515 in different cutting stages. Changes of allele numbers in samples of different cutting stages were observed in seven and six EstSSR loci of the RB72454 and RB867515 varieties, respectively. Reduction of allele numbers was observed in the samples collected in the fourth and sixth cutting stages of the RB72454 variety. In contrast, an increase of the allele numbers was detected in the samples collected on fourth, sixth, and seventh cutting stages of the RB867515 variety. Unchanged allele numbers were observed only in EstB41, EstC84, and EstB130 loci of the RB72454 variety, and EstB41, EstC67, EstA68, and EstB130 loci of the RB867515 variety. The variety RB867515 has lower polymorphism and values of HO than the RB72454 variety in different stages of cutting. At molecular level, in Est-SSR loci, the RB72454 variety showed higher changes in subsequent stages of cutting than RB867515. The similarities and divergences at molecular level between varieties RB72454 and RB867515 observed in the 10 Est-SSR loci during subsequent cutting stages can not explain the reduced productivity frequently observed after subsequent cutting stages but showed that phenotypic and physiological changes after each cutting stage are also accompanied by changes at genomic level.
Assuntos
Etiquetas de Sequências Expressas , Repetições de Microssatélites , Saccharum/genética , Alelos , DNA de Plantas/genética , Variação Genética , Heterozigoto , Polimorfismo Genético , Reprodução/genéticaRESUMO
There is evidence that regulation of follicle selection in cattle involves locally produced growth factors. In the present study, we investigated the expression of members of the fibroblast growth factor (FGF) 7 family during follicle deviation. The largest and second largest follicles were recovered during the second day of a synchronised follicle wave and the future dominant and future subordinate follicles were identified based on diameter and cytochrome P450, family 19, subfamily A, polypeptide 1 (CYP19A1) mRNA levels in granulosa cells. Theca cells of the future dominant follicle contained less mRNA encoding FGF7 and FGF10 compared with those from the future subordinate follicle 2.5 days after ovulation, before a significant difference between the diameters of the future dominant and future subordinate follicles could be observed, but FGF22 mRNA levels did not change. Levels of mRNA encoding FGF receptors FGFR1B and FGFR2B in theca and granulosa cells, respectively, were lower in the future dominant follicle compared with the future subordinate follicle. Addition of FGF10 to granulosa cells in vitro significantly decreased oestradiol secretion, as well as CYP19A1, FSH receptor (FSHR) and insulin-like growth factor 1 receptor (IGF1R) mRNA abundance, whereas FGF22 had no effect. We conclude that FGF10 and FGFR2B expression is increased in the future subordinate follicle before morphological deviation, which may contribute to follicle selection.
Assuntos
Fator 10 de Crescimento de Fibroblastos/metabolismo , Folículo Ovariano/metabolismo , Ovário/metabolismo , Ovulação/metabolismo , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/metabolismo , Animais , Bovinos , Feminino , Fator 10 de Crescimento de Fibroblastos/farmacologia , Hormônio Foliculoestimulante/metabolismo , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Folículo Ovariano/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ovulação/efeitos dos fármacos , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Receptores do FSH/metabolismo , Células Tecais/efeitos dos fármacos , Células Tecais/metabolismoRESUMO
We analyzed 80 plants of the sugarcane (Saccharum spp) variety 'RB867515' in order to investigate its diversity and genetic structure at the molecular level. Four simple sequence repeat (SSR) loci (UGSM51, SMC1237, SEGMS1069, and UGSM38) and five expressed sequence tag (EST)-SSR loci (ESTA68, ESTB92, ESTB145, ESTC66, and ESTC84) were used as molecular markers. The polymorphic loci rate was 66.6%. A total of 17 alleles and an average of 1.88 alleles/locus were detected. The number of alleles in the EST-SSR loci was lower than the number of alleles in the SSRs of non-expressed loci. The mean observed heterozygosity among the nine SSR loci was 0.3291. Genetic structure analysis showed that 'RB867515' contains alleles from three ancestral groups (K = 3), but there is little admixing of alleles in the same plant (from 0.8 to 17.3%); only 1.88% of the plants shared alleles from two or three groups. ESTB92, ESTC84, and UGSM38 were monomorphic, but there was evidence of polymorphism in ESTA68, ESTB145, ESTC66, UGSM51, SMC1237, and SEGMS1069, indicating that 'RB867515' has variability at the molecular level and the potential to be used as a parent in breeding programs. The molecular variability observed in 'RB867515' indicates that the clone terminology that is used to identify this cultivar is inconsistent with the original meaning of "clone", which is defined as a sample of genetically identical plants.
Assuntos
Variação Genética , Saccharum/classificação , Saccharum/genética , DNA de Plantas/análise , Evolução Molecular , Etiquetas de Sequências Expressas , Genoma de Planta , Repetições de Microssatélites , FilogeniaRESUMO
BACKGROUND: Renal transplantation is the treatment of choice for patients with stage V chronic kidney disease, which does not have contraindications to the procedure and is more cost-effective than dialysis treatments and provides better survival and quality of life. OBJECTIVE: The objective of this study was to evaluate the incidence of postoperative complications in kidney transplant recipients in a reference hospital. METHODOLOGY: This was a descriptive and retrospective study involving the analysis of patient records during hospitalization and outpatient treatment. We analyzed the demographics, clinical indicators, surgical techniques, and postoperative complications. RESULTS: In the analysis of 147 transplantations, there was a higher incidence of transplantation in female recipients, average age of 37 years with a predominance of cadaveric transplantation. Of all pretransplantation comorbidities, hypertension was the most frequent. The overall incidence of surgical complications was 29.9%, with an incidence of vascular complications of 12.7%, 13.4% of surgical site complications, 8.2% of urologic complications, and 3% of hemorrhagic complications. DISCUSSION: Vascular complications are serious complications and are associated with increased risk of graft loss (relative risk, 8.4), particularly arterial thrombosis. Patients with ureteral anastomosis using Lich-Gregoir technique showed lower urologic complications compared with patients with anastomosis by Leadbetter-Politano technique. CONCLUSION: Surgical complications have different clinical effects, depending on their category. The vascular complications are associated with graft lost.
Assuntos
Anastomose Cirúrgica/métodos , Falência Renal Crônica/cirurgia , Transplante de Rim/métodos , Complicações Pós-Operatórias/epidemiologia , Trombose/epidemiologia , Abscesso/epidemiologia , Adulto , Transfusão de Sangue , Brasil/epidemiologia , Feminino , Humanos , Incidência , Hérnia Incisional/epidemiologia , Linfocele/epidemiologia , Masculino , Hemorragia Pós-Operatória/epidemiologia , Hemorragia Pós-Operatória/terapia , Qualidade de Vida , Diálise Renal , Reoperação , Estudos Retrospectivos , Infecção da Ferida Cirúrgica/epidemiologia , Transplantados , Ureter/cirurgia , Obstrução Ureteral/epidemiologia , Fístula Urinária/epidemiologiaRESUMO
Retrotransposons are abundant in the genomes of plants. In the present study, inter-retrotransposon amplified polymorphism (IRAP) and retrotransposon-microsatellite amplified polymorphism (REMAP) markers were developed for the cassava genome (Manihot esculenta Crantz). Four cassava cultivars (Fécula Branca, IPR-União, Olho Junto, and Tamboara, two samples per cultivar) were used to obtain IRAP and REMAP fingerprints. Twelve designed primers were amplified alone and in combinations. The 42 IRAP/REMAP primer combinations amplified 431 DNA segments (bands; markers) of which 36 (8.36%) were polymorphic. The largest number of informative markers (16) was detected using the primers AYF2 and AYF2xAYF4. The number of bands for each primer varied from 3 to 16, with an average of 10.26 amplified segments per primer. The size of the amplified products ranged between 100 and 7000 bp. The AYF2 primer generated the highest number of amplified segments and showed the highest number of polymorphic bands (68.75%). Two samples of each cassava cultivar were used to illustrate the usefulness and the polymorphism of IRAP/REMAP markers. IRAP and REMAP markers produced a high number of reproducible bands, and might be informative and reliable for investigation of genetic diversity and relationships among cassava cultivars.
Assuntos
Manihot/genética , Repetições de Microssatélites , Polimorfismo Genético , Retroelementos , Marcadores GenéticosRESUMO
BACKGROUND AND PURPOSE: The synthetic peptide PnPP-19 has been studied as a new drug candidate to treat erectile dysfunction. However, PnTx2-6, the spider toxin from which the peptide was designed, induces hyperalgesia. Therefore, we intended to investigate the role of PnPP-19 in the nociceptive pathway. EXPERIMENTAL APPROACH: Nociceptive thresholds were measured by paw pressure test. PnPP-19 was administered intraplantarly alone or with selective cannabinoid or opioid receptor antagonists. The hydrolysis of PnPP-19 by neutral endopeptidase (NEP) (EC 3.4.24.11), an enzyme that cleaves enkephalin, was monitored by HPLC and the cleavage sites were deduced by LC-MS. Inhibition by PnPP-19 and Leu-enkephalin of NEP enzyme activity was determined spectrofluorimetrically. KEY RESULTS: PnPP-19 (5, 10 and 20 µg per paw) induced peripheral antinociception in rats. Specific antagonists of µ opioid receptors (clocinnamox), δ opioid receptors (naltrindole) and CB1 receptors (AM251) partly inhibited the antinociceptive effect of PnPP-19. Inhibition of fatty acid amide hydrolase by MAFP or of anandamide uptake by VDM11 enhanced PnPP-19-induced antinociception. NEP cleaved PnPP-19 only after a long incubation, and Ki values of 35.6 ± 1.4 and 14.6 ± 0.44 µmol·L(-1) were determined for PnPP-19 and Leu-enkephalin respectively as inhibitors of NEP activity. CONCLUSIONS AND IMPLICATIONS: Antinociception induced by PnPP-19 appears to involve the inhibition of NEP and activation of CB1, µ and δ opioid receptors. Our data provide a greater understanding of the antinociceptive effects of PnPP-19. This peptide could be useful as a new antinociceptive drug candidate.
Assuntos
Analgésicos Opioides/farmacologia , Inibidores Enzimáticos/farmacologia , Neprilisina/antagonistas & inibidores , Peptídeos/farmacologia , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptores Opioides/metabolismo , Venenos de Aranha/química , Animais , Relação Dose-Resposta a Droga , Masculino , Neprilisina/metabolismo , Ratos , Ratos Wistar , Relação Estrutura-AtividadeRESUMO
Amplified fragment length polymorphism (AFLP) analysis was used to evaluate DNA polymorphism in Pilosocereus gounellei with the aim of differentiating samples grown in different Brazilian semiarid regions. Seven primer pairs were used to amplify 703 AFLP markers, of which 700 (99.21%) markers were polymorphic. The percentage of polymorphic markers ranged from 95.3% for the primer combination E-AAG/M-CTT to 100% for E-ACC/M-CAT, E-ACC/M-CAA, E-AGC/M-CAG, E-ACT/M-CTA, and E-AGG/M-CTG. The largest number of informative markers (126) was detected using the primer combination E-AAC/M-CTA. Polymorphism of the amplified DNA fragments ranged from 72.55% (in sample from Piauí State) to 82.79% (in samples from Rio Grande Norte State), with an average of 75.39%. Despite the high genetic diversity of AFLP markers in xiquexique, analysis using the STRUCTURE software identified relatively homogeneous clusters of xiquexique from the same location, indicating a differentiation at the molecular level, among the plant samples from different regions of the Caatinga biome. The AFLP methodology identified genetically homogeneous and contrasting plants, as well as plants from different regions with common DNA markers. Seeds from such plants can be used for further propagation of plants for establishment of biodiversity conservation units and restoration of degraded areas of the Caatinga biome.
Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Cactaceae/genética , Ecossistema , Marcadores Genéticos , Genética Populacional , Polimorfismo Genético , Brasil , Variação GenéticaRESUMO
The time at which follicles acquire LHR in bovine granulosa cells is the subject of some controversy among researchers. The main objective of the present study was to assess the mRNA expression of LHR and LRBP (mRNA protein binding), a post-transcriptional suppressor of LHR mRNA expression, in granulosa cells from the two largest follicles around the expected time of follicle deviation in Nelore heifers. First, the interval between ovulation and follicle deviation in 20 Nelore heifers was determined (2.3 ± 0.2 days after ovulation). Ovulation was hormonally synchronized, and then, heifers were slaughtered on days 2, 2.5 and 3 after ovulation (before, during and after, respectively, the expected time of follicle deviation), and granulosa cells from the two largest follicles were collected. The mRNA abundance of an LHR fragment common to all isoforms (total LHR) and LRBP was assessed by real-time RT-PCR, and LHR alternative transcripts were assessed by semiquantitative RT-PCR followed by electrophoresis. LHR mRNA expression was not detected before the expected time of deviation. Total LHR mRNA abundance was greater in the largest follicle and increased from day 2.5 to 3. In contrast, LRBP mRNA was detected starting on day 2 and was more expressed in the second largest follicle on days 2.5 and 3. The present data suggest that the expression of LHR mRNA in bovine granulosa cells is established after follicle deviation and that the lower abundance of LRBP mRNA after the expected time of deviation may contribute to greater expression of LHR in the bovine dominant follicle.
Assuntos
Bovinos/genética , Células da Granulosa/metabolismo , Folículo Ovariano/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Receptores do LH/genética , Animais , Feminino , Expressão Gênica , Ovulação/genética , RNA Mensageiro/genética , Transdução de Sinais/genéticaRESUMO
In this study, we analyzed dominant molecular markers to estimate the genetic divergence of 26 popcorn genotypes and evaluate whether using various dissimilarity coefficients with these dominant markers influences the results of cluster analysis. Fifteen random amplification of polymorphic DNA primers produced 157 amplified fragments, of which 65 were monomorphic and 92 were polymorphic. To calculate the genetic distances among the 26 genotypes, the complements of the Jaccard, Dice, and Rogers and Tanimoto similarity coefficients were used. A matrix of Dij values (dissimilarity matrix) was constructed, from which the genetic distances among genotypes were represented in a more simplified manner as a dendrogram generated using the unweighted pair-group method with arithmetic average. Clusters determined by molecular analysis generally did not group material from the same parental origin together. The largest genetic distance was between varieties 17 (UNB-2) and 18 (PA-091). In the identification of genotypes with the smallest genetic distance, the 3 coefficients showed no agreement. The 3 dissimilarity coefficients showed no major differences among their grouping patterns because agreement in determining the genotypes with large, medium, and small genetic distances was high. The largest genetic distances were observed for the Rogers and Tanimoto dissimilarity coefficient (0.74), followed by the Jaccard coefficient (0.65) and the Dice coefficient (0.48). The 3 coefficients showed similar estimations for the cophenetic correlation coefficient. Correlations among the matrices generated using the 3 coefficients were positive and had high magnitudes, reflecting strong agreement among the results obtained using the 3 evaluated dissimilarity coefficients.
Assuntos
Variação Genética , Genótipo , Zea mays/genética , Análise por Conglomerados , Marcadores Genéticos , Polimorfismo Genético , Técnica de Amplificação ao Acaso de DNA Polimórfico , Zea mays/classificaçãoRESUMO
The purpose of this study was to analyze the genetic diversity of 15 sugary-1 sweet corn lines by microsatellite markers. One hundred pairs of simple sequence repeat primers that were mapped for field corn were tested. Of these primers, 15% were polymorphic, and all were selected for the evaluation. These primers identified a total of 39 alleles among the 15 loci that were evaluated. The number of alleles per locus in the genotypes ranged from 2 to 4, with an average of 2.60 alleles per locus; the highest number of alleles was observed at the loci Bnlg1083, Umc1241, and Umc1590. The occurrence of null alleles at locus Umc1363 was evident only in line DN44. The proportion of polymorphic loci was the highest in lines DN17.1 and DN6 (73.33%), whereas lines DN47, DN23, and DN28 were more monomorphic than other lines. The loci Bnlg1083 and Umc1506 were polymorphic in 8 and 7 lines, respectively, indicating that these loci might be effective and promising for the identification of polymorphism in other sweet corn lines. The genetic diversity calculated by Rogers' genetic distances indicated the lowest genetic similarity between lines DN9 and DN28 (0.7603) and the highest similarity between lines DN19 and DN6 (0.3724). The dendrogram obtained by the unweighted pair-group method based on arithmetic averages indicated the formation of 4 major groups, showing the crossing of the genotypes DN19 and DN6 with DN8 as a possible alternative for the expression of heterozygosis.
Assuntos
Variação Genética , Repetições de Microssatélites/genética , Zea mays/genética , Alelos , Deriva Genética , Genótipo , Polimorfismo Genético , Especificidade da EspécieRESUMO
In this study, we measured the genetic diversity within and among a set of 9 commercial sugarcane varieties used for alcohol and sugar production using 17 microsatellite DNA markers. The UGSM148 and UGSM59 primers were monomorphic for all 74 sugarcane samples. The estimated proportion of simple sequence repeated (SSR) polymorphic loci was 88.23%; 17 alleles were detected. The mean gene diversity of all SSR loci was 0.7279. The highest observed heterozygosity (HO) value was found in the RB72454 variety, whereas the lowest HO value was recorded in the SP813250 variety. The SP813250, RB845210, and RB835054 sugarcane varieties were the most genetically uniform varieties. An extremely high level of population differentiation was detected in the varieties exhibiting similar agronomic characteristics. Analysis of the genetic structure of the 9 sugarcane varieties using SSR markers was especially important to identify SSR loci with high levels of heterozygosity and to identify varieties showing the highest levels of heterozygosity. The monomorphic primers may be used to evaluate the genetic stability of sugarcane during cycles of vegetative multiplication, i.e., propagation via rhizomes.
Assuntos
DNA de Plantas/genética , Saccharum/classificação , Saccharum/genética , Primers do DNA/genética , Variação Genética , Instabilidade Genômica , Heterozigoto , Repetições de Microssatélites , FilogeniaRESUMO
The LH plays a key role in controlling physiological processes in the ovary acting via LH receptor (LHR). In general, the effects of LHR on the regulation of granulosa cell differentiation are mediated mainly via the Gs-protein/adenylyl cyclase/cAMP system; however, the LHR activation could also induce phospholipase C (PLC)/inositol trisphosphate (IP3) via Gq/11 system. Additionally, the expression of G-proteins (GNAS, GNAQ, and GNA11) and PLC ß has been showed in bovine antral follicle, concomitant with an increase in LHR expression. To gain insight into the effects of superstimulation with FSH (P-36 protocol) or FSH combined with equine chorionic gonadotropin (eCG; P-36/eCG protocol) on the mRNA expression of proteins involved in LHR signaling in bovine granulosa cells, Nelore cows (Bos indicus) were treated with two superstimulatory protocols: P-36 protocol or P-36/eCG protocol (replacement of the FSH by eCG administration on the last day of treatment). Nonsuperstimulated cows were only submitted to estrous synchronization without ovarian superstimulation. The granulosa cells were harvested from follicles and mRNA abundance of GNAS, GNAQ, GNA11, PLCB1, PLCB, PLCB4, and adenylyl cyclase isoforms (ADCY3, ADCY4, ADCY6, ADCY8, and ADCY9) was measured by real-time reserve transcription followed by polymerase chain reaction. No differences on mRNA abundance of target genes were observed in granulosa cells of cows submitted to P-36 protocol compared with control group. However, the cows submitted to P-36/eCG protocol showed upregulation on the mRNA abundance of target genes (except ADCY8) in granulosa cells. Although the P-36 protocol did not regulate mRNA expression of the proteins involved in the signaling mechanisms of the cAMP and IP3 systems, the constant presence of GNAS, GNAQ, GNA11, PLCB1, PLCB3, PLCB4, and adenylyl cyclase isoforms (ADCY3, ADCY4, ADCY6, and ADCY9) mRNA and the upregulation of these genes in granulosa cells from cows submitted to P-36/eCG protocol reinforce the participation of Gq/11/PLC/IP3 signaling as well as Gs-protein/adenylyl cyclase/cAMP system on LHR pathways during bovine granulosa cell differentiation submitted to superstimulatory treatments.
Assuntos
Bovinos/metabolismo , Gonadotropina Coriônica/farmacologia , Hormônio Foliculoestimulante/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Indução da Ovulação/veterinária , Receptores do LH/metabolismo , Animais , Feminino , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Cavalos , Indução da Ovulação/métodos , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
In the mammalian ovary, FGF10 is expressed in oocytes and theca cells and is a candidate for paracrine signaling to the developing granulosa cells. To gain insight into the participation of FGF10 in the regulation of fetal folliculogenesis, we assessed mRNA expression patterns of FGF10 and its receptors, FGFR1B and FGFR2B, in relation to fetal follicle dynamics and localized FGF10 protein in bovine fetal ovaries at different ages. Primordial, primary, secondary, and antral follicles were first observed on Days 75, 90, 150, and 210 of gestation, respectively. The levels of GDF9 and BMP15 mRNA, markers for primordial and primary follicles, respectively, increased during fetal ovary development in a consistent manner with fetal follicle dynamics. CYP17A1 mRNA abundance increased from Day 60 to Day 75 and then from Day 120 to Day 150, coinciding with the appearance of secondary follicles. FGF10 mRNA abundance increased from Day 90, and this increase was temporally associated with increases in FGFR1B mRNA abundance and in the population of primary follicles. In contrast, FGFR2B mRNA expression was highest on Day 60 and decreased thereafter. FGF10 protein was localized to oogonia and oocytes and surrounding granulosa cells at all fetal ages. The present data suggest a role for FGF10 in the control of fetal folliculogenesis in cattle.
Assuntos
Bovinos/embriologia , Fator 10 de Crescimento de Fibroblastos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Ovário/embriologia , Animais , Feminino , Fator 10 de Crescimento de Fibroblastos/genética , Ovário/metabolismo , Receptores de Fatores de Crescimento de Fibroblastos/genética , Receptores de Fatores de Crescimento de Fibroblastos/metabolismoRESUMO
This study used esterases and ribosomal DNA (rDNA) markers to determine endophytic variability in order to better understand endophyte-host interactions. Polyacrylamide gel electrophoresis and esterase isoenzymes (EST; EC 3.1.1.3), with α-naphthyl acetate and ß-naphthyl acetate as substrates, were used to assess relationships among endophytes. ITS1-5.8S-ITS2 sequencing data were used as rDNA markers. Thirty-two esterases were obtained from 37 isolates of Saccharum spp, which clustered into five endophyte groups. Esterase EST-06 was observed with the highest frequency, being present in 22 of the 37 isolates analyzed, followed by esterase EST-11, which was present in 20 isolates. The esterases EST-10 and EST-14 were present in 19 isolates and EST-09 was present in 18 isolates. The esterase EST-01 was unique to isolate 33 and can, therefore, be used as a marker for this isolate. None of the esterases identified were common to all isolates tested. Similarly, phylogenetic analysis, based on rDNA sequence data, classified the isolates into 5 genus groups: 1) Curvularia with a 100% bootstrap value (BP), 2) Alternaria with 100% BP, 3) Epicoccum with 60% BP, 4) Phoma with 89% BP, and 5) Saccharicola with 100% BP. This polyphyletic analysis based on several markers, therefore, proved to be a valuable approach in determining the relationship between variation in endophytes and their associated host plants. Furthermore, both the esterase and rDNA analyses obtained similar results and were equally effective in resolving relationships.
Assuntos
Endófitos/classificação , Endófitos/isolamento & purificação , Esterases/genética , Saccharum/microbiologia , Ascomicetos/classificação , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/isolamento & purificação , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Endófitos/genética , Marcadores Genéticos , Variação Genética , Estrutura Molecular , Naftóis/química , Filogenia , Folhas de Planta/microbiologia , Análise de Sequência de DNARESUMO
Genetic diversity and structure were analyzed in 10 accessions belonging to Banco Ativo de Germoplasma de Capsicum located at Federal University of Piauí in northwestern Brazil that receives pepper samples grown in community gardens in various regions and Brazilian states. Selections were made from seeds of C. chinense (4 accessions), C. annuum (5 accessions), and C. baccatum (1 accession). Samples consisting of leaves were collected from 4-10 plants of each accession (a total of 85 plants). Native polyacrylamide gel electrophoresis was used to identify α- and ß-esterase polymorphisms. Polymorphism was clearly detected in 5 loci. Sixteen alleles were found at 5 α/ß-esterase loci of the three Capsicum species. In the C. chinense samples, the highest HO and HE values were 0.3625 and 0.4395, respectively, whereas in C. annuum samples, HO and HE values were 0.2980 and 0.3310, respectively; the estimated HO and HE values in C. chinense samples were higher than those detected in C. annuum samples. A deficit of homozygous individuals was found in C. chinense (FIS = -0.6978) and C. annuum (FIS = 0.7750). Genetic differentiation between C. chinense and C. annuum at these loci was high (FST = 0.1867) indicating that C. chinense and C. annuum are genetically structured species for α/ß- esterase isozymes. The esterase analysis showed high genetic diversity among the C. chinense and C. annuum samples and very high genetic differentiation (FST = 0.6321) among the C. chinense and C. annuum samples and the C. baccatum accession.
Assuntos
Capsicum/genética , Esterases/genética , Polimorfismo Genético , Alelos , Capsicum/metabolismo , Esterases/classificação , Esterases/metabolismo , Loci Gênicos , Genótipo , Isoenzimas , Fenótipo , FilogeniaRESUMO
Currículos de medicina veterinária devem disponibilizar ferramentas para que os futuros profissionais atendam a demanda da sociedade, que inclui preocupações diretas com os animais. O objetivo deste trabalho foi traçar um perfil do panorama geral do ensino da medicina veterinária em relação a questões de bem-estar e dor animal. O método utilizado foi análise de documentos disponíveis online e coleta de dados via aplicação de questionários para coordenadores de curso de medicina veterinária. A descrição do curso, sua grade curricular e ementário foram estudados. Observou-se que 46% das 94 instituições estudadas apresentavam a disciplina de bem-estar animal e 26% ofereciam a disciplina de etologia. Houve evidência de que há uma pronta relação com a esfera física do bem-estar animal, sendo que as outras duas esferas, comportamental e psicológica, não recebem atenção similar ao longo dos cursos. Na avaliação do ementário, o termo "bem-estar animal" é empregado com caráter difuso e o termo "dor" encontra-se presente em 54% dos cursos estudados, relacionado principalmente a disciplinas de patologia, fisiologia, farmacologia e anestesiologia. Conclui-se que o ensino brasileiro de medicina veterinária enfatiza a esfera física do bem-estar animal, sendo importante o enriquecimento em relação às esferas comportamental e psicológica e ao ensino da dor.
The curricula of veterinary medicine should provide tools for future professionals to meet society demands, which include direct concerns for the animals. The overall scenario of education in veterinary medicine on issues of animal welfare and pain was studied. This study was conducted through the analysis of documents available online and via questionnaires to coordinators of veterinary medicine programs. The program description, its curriculum and course content descriptions were considered. Results show that 46% of the 94 institutions studied offer an animal welfare course and 26% offer an ethology course. We observed a direct relationship with the physical component of animal welfare; the other two components, the behavioral and psychological ones, do not receive similar attention throughout the programs. In the study of course contents of the veterinary programs, the term 'animal welfare' is used in a diffuse manner and the term 'pain' appears in 54% of the programs studied, mainly related to disciplines covering its pathology, physiology, pharmacology and anesthesiology. We conclude that in the teaching of veterinary medicine in Brazil there is an emphasis on the physical realm of animal welfare, and that there is room for improvement in the naturalness and psychological realms and in the teaching of pain.
Assuntos
Animais , Bem-Estar do Animal/estatística & dados numéricos , Educação em Veterinária/estatística & dados numéricos , Educação em Veterinária/métodosRESUMO
Os mecanismos que coordenam o desenvolvimento folicular ainda não são completamente conhecidos e, portanto, constituem o alvo de numerosas investigações, seja por facilitarem a compreensão da fisiologia ou por serem promissoras ferramentas para a reprodução assistida. Recentemente, diversos peptídeos ovarianos de ação local têm sido descritos por participarem do controle de todas as fases do desenvolvimento folicular, bem como da modulação de hormônios esteróides ovarianos e gonadotrofinas; entre esses peptídeos estão os fatores de crescimento fibroblástico (FGFs). Os FGFs têm sido extensamente investigados em diversas fases do desenvolvimento folicular e parecem controlar processos como atresia folicular, esteroidogênese, bem como o desenvolvimento folicular pré-antral, sendo a subfamília do FGF7 uma das mais investigadas neste contexto. Assim, esta revisão tem como objetivo sumarizar a participação da subfamília do FGF7 no controle da foliculogênese antral de bovinos.
The mechanisms that coordinate follicular development are not well known and are, therefore, target of numerous investigations for facilitateing the understanding of physiology or as promising tools for assisted reproduction. Recently, several ovarian peptides with local action have been reported to participate in the control of follicular development in all stages and modulation of gonadotropins andovarian steroid hormones. In this context, fibroblast growth factors (FGFs) have been extensively investigated in different stages of follicular development and seem to control processes such as follicular atresia, steroidogenesis, and pre-antral follicle development, where FGF7 subfamily is one of the most investigated. Therefore, the objective of this review is to summarize the participation of the FGF7 subfamily in the control of bovine antral folliculogenesis.