RESUMO
Wisent, or European bison (Bison bonasus), is listed as "vulnerable" on the IUCN Red List of Threatened Species and is therefore protected by international law. For the first time, a Wisent embryo has been obtained in vitro. This procedure creates a new opportunity to protect and increase Wisent reproductive potential and thereby opens new possibilities for the establishment of a controlled and broad reserve of the gene pool.
Assuntos
Bison/fisiologia , Técnicas de Cultura Embrionária/veterinária , Fertilização in vitro/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Animais , Blastocisto , Chlorocebus aethiops , Técnicas de Cocultura , Técnicas de Cultura Embrionária/métodos , Espécies em Perigo de Extinção , Feminino , Fertilização in vitro/métodos , Técnicas de Maturação in Vitro de Oócitos/métodos , Masculino , Células VeroRESUMO
UNLABELLED: Observational studies have suggested that statins may have beneficial effects on outcomes in chronic obstructive pulmonary disease (COPD) patients. These effects may be mediated through an anti-inflammatory effect of statins. The purpose of this pilot-study was to determine whether statins have an anti-inflammatory effect on the lungs of COPD patients. We conducted randomized, controlled, parallel group pilot-study to compare the effects of atorvastatin (n=12) or placebo (n=6) on lung inflammation in patients with mild to moderate COPD. The primary endpoint was change in CD45+ cells expression measured by immunohistochemistry and changes in expression of genes measured using microarrays in lung biopsy (TBB) samples before and after 12 weeks of treatment with atorvastatin 40 mg/day. All subjects had spirometry, lung volumes, diffusing capacity of the lungs for carbon monoxide (DLCO), St George's Respiratory Questionnaire (SGRQ), 6 minute walk distance (6 MWD), serum lipids, hs-CRP, induced sputum (IS), bronchoscopy and TBB carried out at baseline and after treatment. TBB specimens were processed for histology, immunohistochemistry and genome-wide association studies (GWAS) profiling. Seventeen subjects completed the study. There was a significant improvement in SGRQ with mean SGRQ decreased by 12 points after treatment with atorvastatin (P=0.012). Atorvastatin treatment produced a significant 34% reduction in sputum neutrophil count, and a 57% reduction in CD45+ cells in lung biopsies (expressed as integrated optical density -IOD; median IOD 62.51% before, 27.01% after atorvastatin treatment, P=0.008). In patients' lung tissue atorvastatin treatment produced downregulation of key genes involved in inflammatory processes, immune response, and leukocyte activation. These data demonstrate the pulmonary anti-inflammatory effects of atorvastatin in COPD patients with the potential for beneficial clinical effects. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01748279.
Assuntos
Anti-Infecciosos/uso terapêutico , Atorvastatina/uso terapêutico , Pulmão/efeitos dos fármacos , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Idoso , Biópsia , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Marcadores Genéticos , Estudo de Associação Genômica Ampla , Humanos , Imuno-Histoquímica , Mediadores da Inflamação/imunologia , Pulmão/imunologia , Pulmão/fisiopatologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Polônia , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/genética , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Recuperação de Função Fisiológica , Testes de Função Respiratória , Índice de Gravidade de Doença , Método Simples-Cego , Fatores de Tempo , Resultado do TratamentoRESUMO
The objective of this study was to examine the feasibility of identification and selection of cattle embryos based on green fluorescence (GFP-positive) in order to obtain calves carrying an integrated transgene. The construct used (pbLGTNF-EGFP) contained the human tumor necrosis factor alpha (hTNFalpha) gene fused to the bovine beta-lactoglobulin promoter (bLG) in plasmid vector pCX-EGFP. In four experiments, 76 zygotes were injected; eight of them developed to the morulae/blastocysts stage of which only five were GFP positive (one of them 100%, one-50%, three- 25%). All of the GFP positive embryos were transferred to recipients. Two calves were born: one after transfer of the 100% GFP positive embryo and the other after transfer of one of the 25% GFP positive embryos. Both animals were healthy with normal weight when compared to two control calves. The integration of pbLGTNF-EGFP in the host genome could not be detected in either of the calves, suggesting that GFP is an unreliable marker for preimplantation screening of embryos.
Assuntos
Animais Geneticamente Modificados/genética , Biomarcadores , Bovinos/genética , Embrião de Mamíferos/citologia , Proteínas de Fluorescência Verde , Animais , Blastocisto , Transferência Embrionária/veterinária , Embrião de Mamíferos/metabolismo , Estudos de Viabilidade , Feminino , Fertilização in vitro/veterinária , Vetores Genéticos , Lactoglobulinas/genética , Microinjeções/veterinária , Plasmídeos/genética , Reação em Cadeia da Polimerase/veterinária , Proteínas Recombinantes de Fusão/genética , Transgenes/genética , Transplante/veterinária , Fator de Necrose Tumoral alfa/genéticaRESUMO
The objective of this study was to test a new co-culture system of bovine embryos, which we call "mixed co-culture." This system consists of culturing embryos on cell monolayers composed of both Vero and BRL cells (Vero/BRL). Cumulus-oocyte complexes from ovaries of slaughtered cows were matured and fertilized in vitro. The presumptive zygotes were cultured with Vero/BRL (Group 1), BRL (Group 2) or Vero (Group 3) cell monolayers, in 40 microL drops of Menezo B2 medium supplemented with 10% FBS and antibiotics. The development of the presumptive zygotes was compared on Day 2 [48 h post insemination (pi)] and Day 7 (168 h pi). On Day 2, there was no difference between the groups. On Day 7, the highest percentage of compacted morulae/blastocysts was observed in mixed co-culture of Vero/BRL cells: 40% versus 36% on BRL versus 27% on Vero cell monolayers. The differences were statistically significant (P < or = 0.05). Among compacted morulae/blastocysts, blastocysts prevailed in mixed co-culture: 67% on Vero/BRL as compared with 55% on BRL and 27% on Vero cell monolayers. The differences were highly statistically significant (P < or = 0.01). The results suggest that Vero/BRL cells improve the development of bovine embryos.
Assuntos
Bovinos/embriologia , Técnicas de Cocultura/veterinária , Reprodução/fisiologia , Zigoto/crescimento & desenvolvimento , Animais , Bovinos/fisiologia , Linhagem Celular , Chlorocebus aethiops , Técnicas de Cocultura/métodos , Feminino , Fertilização in vitro/veterinária , Masculino , Células VeroRESUMO
The role of persistent chlorinated hydrocarbons (PCH) in the reproductive disorders in ruminants is not well documented. In the present study we have examined the effect of these compounds and their metabolites on the secretion of estradiol (E2) and progesterone (P4) by bovine granulosa cells in vitro. Granulosa cells were isolated from large follicles (> or = 8 mm diameter) by gently washing the internal follicle wall. Aliquots of approximately 4 X 10(5) viable granulosa cells in 0.5 ml medium were cultured at 37 degrees C in an atmosphere of 5% CO2 and 95% air. Granulosa cells were cultured for 96h in a medium containing different concentrations (10(-1)-10(-4) ng/ml) of a PCH combination. Estradiol and progesterone were measured in unextracted granulosa cell culture medium by Enzyme Immunoassay (EIA). The exposure of granulosa cells to a combination of these organochlorine compounds in vitro results in a slight decrease of estradiol secretion only at the highest studied concentration of the PCH combination. However, the secretion of progesterone by these cells was seriously decreased, even by concentrations found in ovaries from animals kept under natural environmental conditions. The in vitro culture system of granulosa cells from preovulatory follicles may be useful in screening toxic effects of pesticides in animal reproduction.