RESUMO
In the past two decades, average litter size (ALS) in Entlebucher Mountain dogs decreased by approximately 0.8 puppies. We conducted a GWAS for ALS using the single-step methodology to take advantage of 1632 pedigree records, 892 phenotypes and 372 genotypes (173 662 markers) for which only 12% of the dogs had both phenotypes and genotypes available. Our analysis revealed associations towards the growth differentiation factor 9 gene (GDF9), which is known to regulate oocyte maturation. The trait heritability was estimated at 43.1%, from which approximately 15% was accountable by the GDF9 locus alone. Therefore, markers flanking GDF9 explained approximately 6.5% of the variance in ALS. Analysis of WGSs revealed two missense substitutions in GDF9, one of which (g.11:21147009G>A) affected a highly conserved nucleotide in vertebrates. The derived allele A was validated in 111 dogs and shown to be associated with decreased ALS (-0.75 ± 0.22 puppies per litter). The variant was further predicted to cause a proline to serine substitution. The affected residue was immediately followed by a six-residue deletion that is fixed in the canine species but absent in non-canids. We further confirmed that the deletion is prevalent in the Canidae family by sequencing three species of wild canids. Since canids uniquely ovulate oocytes at the prophase stage of the first meiotic division, requiring maturation in the oviduct, we conjecture that the amino acid substitution and the six-residue deletion of GDF9 may serve as a model for insights into the dynamics of oocyte maturation in canids.
Assuntos
Cães/genética , Fator 9 de Diferenciação de Crescimento/genética , Tamanho da Ninhada de Vivíparos/genética , Mutação de Sentido Incorreto , Sequência de Aminoácidos , Animais , Cruzamento , Feminino , Estudos de Associação Genética/veterinária , Genótipo , Masculino , Linhagem , FenótipoRESUMO
Post-translational modifications of histones, such as acetylation, are involved in regulating chromatin remodelling and gene expression. Proper in vitro maturation (IVM) of canine oocytes, for many reasons, is up to now inefficient. This study aimed to evaluate the post-translational histone H4 acetylation at lysine 5 (H4K5) in immature and post-IVM canine oocytes. Oocyte nuclear stage was assessed using Hoechst 33342 staining. Acetylation patterns were determined by indirect immunofluorescence staining of immature and post-IVM oocytes, using an antibody against the acetylated lysine 5 residue on histone 4 (H4K5ac). The experiment was repeated four times, with a total of 7-17 oocytes evaluated per stage. Immunofluorescence signal was quantified using the NIHimagej software. Data were expressed as a percentage of the average fluorescence intensity of the specific antibody over the intensity of DNA, as determined by Hoescht staining. H4K5ac displayed a significantly higher acetylated pattern in immature oocytes (0.97 ± 0.08) when compared to post-IVM oocytes at different nuclear stages. There was a decrease in the fluorescence level of the matured oocytes with the progression of meiosis (GVBD: 0.47 ± 0.06 and MI/MII: 0.35 ± 0.04). Similarly to other domestic species, we hypothesized that post-translational modification of histone acetylation takes place during meiosis of in vitro matured canine oocytes. However, it remains to be investigated whether these changes occur during in vivo maturation.
Assuntos
Histonas/química , Técnicas de Maturação in Vitro de Oócitos , Meiose/fisiologia , Oócitos/fisiologia , Oogênese/fisiologia , Acetilação , Animais , Cães/fisiologia , Feminino , Fertilização in vitro , Técnica Indireta de Fluorescência para AnticorpoRESUMO
Despite extensive efforts, establishment of bovine embryonic stem (ES) cell lines has not been successful. We hypothesized that culture conditions for in vitro-produced (IVP) embryos, the most used source of inner cell mass (ICM) to obtain ES cells, might affect their undifferentiated state. Therefore, the aim of this work was to improve pluripotency of IVP blastocysts to produce suitable ICM for further culturing. We tested KSR and foetal calf serum (FCS) supplements in SOF medium and ES cell conditioned medium (CM) on IVC (groups: KSR, KSR CM, FCS and FCS CM). Cleavage and blastocyst rates were similar between all groups. Also, embryonic quality, assessed by apoptosis rates (TUNEL assay), total cell number and ICM percentage did not differ between experimental groups. However, expression of pluripotency-related markers was affected. We detected down-regulation of OCT3/4, SOX2 and SSEA1 in ICM of FCS CM blastocysts (p < 0.05). SOX2 gene expression revealed lower levels (p < 0.05) on KSR CM blastocysts and a remarkable variation in SOX2 mRNA levels on FCS-supplemented blastocysts. In conclusion, pluripotency-related markers tend to decrease after supplementation with ES cell CM, suggesting different mechanisms regulating mouse and bovine pluripotency. KSR supplementation did not differ from FCS, but FCS replacement by KSR may produce blastocysts with stable SOX2 gene expression levels.
Assuntos
Bovinos/embriologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Antígenos CD15/metabolismo , Fator 3 de Transcrição de Octâmero/metabolismo , Células-Tronco Pluripotentes/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Animais , Linhagem Celular , Meios de Cultivo Condicionados , Técnicas de Cultura Embrionária/veterinária , Células-Tronco Embrionárias/fisiologia , Fertilização in vitro , Antígenos CD15/genética , Camundongos , Fator 3 de Transcrição de Octâmero/genética , Fatores de Transcrição SOXB1/genéticaRESUMO
Mammalian fetal survival and growth are dependent on a well-established and functional placenta. Although transient, the placenta is the first organ to be formed during pregnancy and is responsible for important functions during development, such as the control of metabolism and fetal nutrition, gas and metabolite exchange, and endocrine control. Epigenetic marks and gene expression patterns in early development play an essential role in embryo and fetal development. Specifically, the epigenetic phenomenon known as genomic imprinting, represented by the non-equivalence of the paternal and maternal genome, may be one of the most important regulatory pathways involved in the development and function of the placenta in eutherian mammals. A lack of pattern or an imprecise pattern of genomic imprinting can lead to either embryonic losses or a disruption in fetal and placental development. Genetically modified animals present a powerful approach for revealing the interplay between gene expression and placental function in vivo and allow a single gene disruption to be analyzed, particularly focusing on its role in placenta function. In this paper, we review the recent transgenic strategies that have been successfully created in order to provide a better understanding of the epigenetic patterns of the placenta, with a special focus on imprinted genes. We summarize a number of phenotypes derived from the genetic manipulation of imprinted genes and other epigenetic modulators in an attempt to demonstrate that gene-targeting studies have contributed considerably to the knowledge of placentation and conceptus development.
Assuntos
Desenvolvimento Fetal/fisiologia , Impressão Genômica/fisiologia , Placenta/metabolismo , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Impressão Genômica/genética , Humanos , GravidezRESUMO
OBJETIVO: Investigar a modulação autonômica exercida sobre o nodo sinusal, por meio da análise da variabilidade da freqüência cardíaca (VFC), em indivíduos jovens e de meia-idade (MI), bem como os efeitos de um programa de treinamento de força resistência sobre tal modulação nos indivíduos de meia-idade. MÉTODO: Trinta e dois homens sadios, sedentários e não-tabagistas, sendo 10 jovens (22,2 ± 1,5 anos) e 22 de MI (49,3 ± 5,3 anos), foram submetidos à aquisição do sinal de eletrocardiograma para análise da VFC no domínio do tempo. Os indivíduos de MI foram divididos em dois grupos: experimental (n= 12) e controle (n= 10). Os indivíduos do grupo experimental foram inseridos em um programa de treinamento de força com duração de três meses. A análise dos dados foi realizada por meio dos testes de Wilcoxon e Mann-Whitney (p< 0,05). RESULTADOS: O grupo MI apresentou redução significativa, em comparação ao jovem, de todas as variáveis utilizadas para a investigação da VFC (SDNN= 33,4 vs. 49,7ms; RMSSD= 29,9 vs. 49,5ms; pNN50= 6,5 vs. 27 por cento). O treinamento promoveu aumento significativo da força e resistência muscular em todos os grupamentos musculares e aumento não significativo das variáveis SDNN (33,4 vs. 37,6ms), RMSSD (30,2 vs. 31,3ms) e pNN50 (7,5 vs. 11,4 por cento). CONCLUSÕES: Os achados deste estudo confirmam que o aumento da idade provoca alteração na modulação autonômica exercida sobre o nodo sinusal, retratada por uma diminuição da VFC em indivíduos de MI, que não foi modificada de maneira significativa pelo tipo de treinamento físico estudado.
OBJECTIVE: To investigate autonomic modulation of the sinus node, by analyzing heart rate variability (HRV) among young and middle-aged individuals, and to assess the effect of an endurance strength training program on this modulation among middle-aged individuals. METHOD: Thirty-two healthy nonsmoking men with sedentary lifestyles, of whom 10 were young (22.2 ± 1.5 years) and 22 were middle-aged (49.3 ± 5.3 years), underwent electrocardiogram signal acquisition for time-domain HRV analysis. The middle-aged individuals were divided into two groups: experimental (n= 12) and control (n= 10). The individuals in the experimental group were enrolled in a strength training program lasting three months. The data analysis was carried out using the Wilcoxon and Mann-Whitney tests (p< 0.05). RESULTS: The middle-aged group presented significant reductions (in relation to the young group) for all the variables used in investigating HRV (SDNN= 33.4 vs. 49.7 ms; RMSSD= 29.9 vs. 49.5 ms; pNN50= 6.5 vs. 27 percent). The training caused a significant increase in muscle strength and endurance for all muscular groups and non-significant increases in the variables SDNN (33.4 vs. 37.6 ms), RMSSD (30.2 vs. 31.3 ms) and pNN50 (7.5 vs. 11.4 percent). CONCLUSIONS: The findings from this study confirm that increased age causes alteration to the autonomic modulation of the sinus node, as demonstrated by reduced HRV in middle-aged individuals, which was not significantly modified by the type of physical training studied.
Assuntos
Adolescente , Pessoa de Meia-Idade , Humanos , Masculino , Sistema Nervoso Autônomo , Exercício Físico , Frequência CardíacaRESUMO
Our aim was to compare the clinical features of panic disorder (PD) patients sensitive to hyperventilation or breath-holding methods of inducing panic attacks. Eighty-five PD patients were submitted to both a hyperventilation challenge test and a breath-holding test. They were asked to hyperventilate (30 breaths/min) for 4 min and a week later to hold their breath for as long as possible, four times with a 2-min interval. Anxiety scales were applied before and after the tests. We selected the patients who responded with a panic attack to just one of the tests, i.e., those who had a panic attack after hyperventilating (HPA, N = 24, 16 females, 8 males, mean age +/- SD = 38.5 +/- 12.7 years) and those who had a panic attack after breath holding (BHPA, N = 20, 11 females, 9 males, mean age +/- SD = 42.1 +/- 10.6 years). Both groups had similar (chi(2) = 1.28, d.f. = 1, P = 0.672) respiratory symptoms (fear of dying, chest/pain discomfort, shortness of breath, paresthesias, and feelings of choking) during a panic attack. The criteria of Briggs et al. [British Journal of Psychiatry, 1993; 163: 201-209] for respiratory PD subtype were fulfilled by 18 (75.0%) HPA patients and by 14 (70.0%) BHPA patients. The HPA group had a later onset of the disease compared to BHPA patients (37.9 +/- 11.0 vs 21.3 +/- 12.9 years old, Mann-Whitney, P < 0.001), and had a higher family prevalence of PD (70.8 vs 25.0%, chi(2) = 19.65, d.f. = 1, P = 0.041). Our data suggest that these two groups--HPA and BHPA patients--may be specific subtypes of PD.
Assuntos
Testes Respiratórios , Hiperventilação/complicações , Transtorno de Pânico/etiologia , Adolescente , Adulto , Ansiedade/complicações , Ansiedade/psicologia , Feminino , Humanos , Hiperventilação/psicologia , Masculino , Pessoa de Meia-Idade , Transtorno de Pânico/diagnósticoRESUMO
Our aim was to compare the clinical features of panic disorder (PD) patients sensitive to hyperventilation or breath-holding methods of inducing panic attacks. Eighty-five PD patients were submitted to both a hyperventilation challenge test and a breath-holding test. They were asked to hyperventilate (30 breaths/min) for 4 min and a week later to hold their breath for as long as possible, four times with a 2-min interval. Anxiety scales were applied before and after the tests. We selected the patients who responded with a panic attack to just one of the tests, i.e., those who had a panic attack after hyperventilating (HPA, N = 24, 16 females, 8 males, mean age ± SD = 38.5 ± 12.7 years) and those who had a panic attack after breath holding (BHPA, N = 20, 11 females, 9 males, mean age ± SD = 42.1 ± 10.6 years). Both groups had similar (chi² = 1.28, d.f. = 1, P = 0.672) respiratory symptoms (fear of dying, chest/pain disconfort, shortness of breath, paresthesias, and feelings of choking) during a panic attack. The criteria of Briggs et al. [British Journal of Psychiatry, 1993; 163: 201-209] for respiratory PD subtype were fulfilled by 18 (75.0 percent) HPA patients and by 14 (70.0 percent) BHPA patients. The HPA group had a later onset of the disease compared to BHPA patients (37.9 ± 11.0 vs 21.3 ± 12.9 years old, Mann-Whitney, P < 0.001), and had a higher family prevalence of PD (70.8 vs 25.0 percent, chi² = 19.65, d.f. = 1, P = 0.041). Our data suggest that these two groups - HPA and BHPA patients - may be specific subtypes of PD.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Adolescente , Testes Respiratórios , Hiperventilação , Transtorno de Pânico , Ansiedade , Transtorno de PânicoRESUMO
Epidemiological and clinical studies have shown a positive correlation between smoking and psychiatric disorders. To investigate the prevalence of cigarette smoking, 277 psychiatric outpatients with anxiety or depressive disorders (DSM-IV) answered a self-evaluation questionnaire about smoking behavior and were compared with a group of 68 control subjects. The diagnoses (N = 262) were: 30.2% (N = 79) major depressive disorder, 23.3% (N = 61) panic disorder, 15.6% (N = 41) social anxiety disorder, 7.3% (N = 19) other anxiety disorders, and 23.7% (N = 62) comorbidity disorders. Among them, 26.3% (N = 69) were smokers, 23.7% (N = 62) were former smokers and 50.0% (N = 131) were nonsmokers. The prevalence of nicotine dependence among the smokers was 59.0% (DSM-IV). The frequency of cigarette smoking did not show any significant difference among the five classes of diagnosis. The social anxiety disorder patients were the heaviest smokers (75.0%), with more unsuccessful attempts to stop smoking (89.0%). The frequency of former smokers was significantly higher among older subjects and nonsmokers were significantly younger (chi2 = 9.13, d.f. = 2, P = 0.01). Our data present some clinical implications suggesting that in our psychiatric outpatient sample with anxiety disorder, major depression and comorbidity (anxiety disorder and major depression), the frequency of cigarette smoking did not differ from the frequency found in the control group or in general population studies. Some specific features of our population (outpatients, anxiety and depressive disorders) might be responsible for these results.
Assuntos
Transtornos Mentais/epidemiologia , Fumar/epidemiologia , Tabagismo/epidemiologia , Adolescente , Adulto , Idoso , Transtornos de Ansiedade/diagnóstico , Transtornos de Ansiedade/epidemiologia , Brasil/epidemiologia , Comorbidade , Transtorno Depressivo Maior/diagnóstico , Transtorno Depressivo Maior/epidemiologia , Estudos Epidemiológicos , Feminino , Humanos , Masculino , Transtornos Mentais/diagnóstico , Pessoa de Meia-Idade , Transtorno de Pânico/diagnóstico , Transtorno de Pânico/epidemiologia , Escalas de Graduação Psiquiátrica , Fumar/psicologia , Tabagismo/psicologiaRESUMO
Epidemiological and clinical studies have shown a positive correlation between smoking and psychiatric disorders. To investigate the prevalence of cigarette smoking, 277 psychiatric outpatients with anxiety or depressive disorders (DSM-IV) answered a self-evaluation questionnaire about smoking behavior and were compared with a group of 68 control subjects. The diagnoses (N = 262) were: 30.2 percent (N = 79) major depressive disorder, 23.3 percent (N = 61) panic disorder, 15.6 percent (N = 41) social anxiety disorder, 7.3 percent (N = 19) other anxiety disorders, and 23.7 percent (N = 62) comorbidity disorders. Among them, 26.3 percent (N = 69) were smokers, 23.7 percent (N = 62) were former smokers and 50.0 percent (N = 131) were nonsmokers. The prevalence of nicotine dependence among the smokers was 59.0 percent (DSM-IV). The frequency of cigarette smoking did not show any significant difference among the five classes of diagnosis. The social anxiety disorder patients were the heaviest smokers (75.0 percent), with more unsuccessful attempts to stop smoking (89.0 percent). The frequency of former smokers was significantly higher among older subjects and nonsmokers were significantly younger (chi² = 9.13, d.f. = 2, P = 0.01). Our data present some clinical implications suggesting that in our psychiatric outpatient sample with anxiety disorder, major depression and comorbidity (anxiety disorder and major depression), the frequency of cigarette smoking did not differ from the frequency found in the control group or in general population studies. Some specific features of our population (outpatients, anxiety and depressive disorders) might be responsible for these results
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Adolescente , Comorbidade , Transtornos Mentais , Fumar , Tabagismo , Transtornos de Ansiedade , Brasil , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Estudos Transversais , Transtorno Depressivo Maior , Transtornos Mentais , Transtorno de Pânico , Prevalência , Escalas de Graduação Psiquiátrica , Estudos Retrospectivos , Fumar , TabagismoRESUMO
OBJECTIVE: To assess the effectiveness of clonazepam, in a fixed dose (2 mg/day), compared with placebo in the treatment of panic disorder patients. METHOD: 24 panic disorder patients with agoraphobia were randomly selected. The diagnosis was obtained using the structured clinical interview for DSM-IV. All twenty-four subjects were randomly assigned to either treatment with clonazepam (2 mg/day) or placebo, during 6 weeks. Efficacy assessments included: change from baseline in the number of panic attacks; CGI scores for panic disorder; Hamilton rating scale for anxiety; and panic associated symptoms scale. RESULTS: At the therapeutic endpoint, only one of 9 placebo patients (11.1%) were free of panic attacks, compared with 8 of 13 (61.5%) clonazepam patients (Fisher exact test; p=0,031). CONCLUSION: the results provide evidence for the efficacy of clonazepam in panic disorder patients.