Evidence for expression and functionality of FSH and LH/hCG receptors in human endometrium.

PURPOSE: Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) mediate intracellular functions by binding their specific protein G-coupled gonadotrophin receptor, respectively FSH receptor (FSHR) and LH/choriogonadotrophin receptor (LHCGR). Whereas the expression of FSHR and LHCGR in mammals was considered gonad-specific and cell-specific, studies identified gonadotrophin receptors in human female extragonadal reproductive tissues. This study aims to demonstrate that gonadotrophin receptors are expressed in endometrium and mediates intracellular functions. METHODS: Collected endometria (n = 12) from healthy patients (mean age of 36 ± 6) were primary cultured for 24 h. The presence of gonadotrophin receptors was evaluated by RT-PCR followed by the sequencing of the resulted amplicons and by immunohistochemistry in original samples. Endometrial primary cultures were treated with increasing concentration (range 0-100 ng/ml) of either recombinant human LH (rhLH) or recombinant human FSH (rhFSH). Endometria controls had gonadotrophin replaced by the same volume of the culture medium. In gonadotrophin-treated samples, it was evaluated the intracellular cyclic adenosine monophosphate (cAMP) content by enzymatic immunoassay and the expression of steroidogenic genes by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). RESULTS: The sequencing of the RT-PCR amplicons confirmed the presence of both gonadotrophin receptors and immunohistochemistry localized them on the membrane of endometrial glands cells throughout the glandular epithelium. The gonadotrophin-receptor complex was able to increase the intracellular cAMP in a dose-response and time-course manner and to induce steroidogenic genes expression. CONCLUSION: This study demonstrates that both gonadotrophin receptors are expressed along the glandular epithelium of endometria and they mediate the effects of gonadotrophins on intracellular functions.

Modulation of gonadotrophin induced steroidogenic enzymes in granulosa cells by d-chiroinositol.

BACKGROUND: d-chiroinositol (DCI) is a inositolphosphoglycan (IPG) involved in several cellular functions that control the glucose metabolism. DCI functions as second messenger in the insulin signaling pathway and it is considered an insulin sensitizer since deficiency in tissue availability of DCI were shown to cause insulin resistance (IR). Polycystic ovary syndrome (PCOS) is a pathological condition that is often accompanied with insulin resistance. DCI can positively affects several aspect of PCOS etiology decreasing the total and free testosterone, lowering blood pressure, improving the glucose metabolism and increasing the ovulation frequency. The purpose of this study was to evaluate the effects of DCI and insulin combined with gonadotrophins namely follicle-stimulating hormone (FSH) and luteinizing hormone (LH) on key steroidogenic enzymes genes regulation, cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and cytochrome P450 side-chain cleavage (P450scc) in primary cultures of human granulosa cells (hGCs). We also investigated whether DCI, being an insulin-sensitizer would be able to counteract the expected stimulator activity of insulin on human granulosa cells (hGCs). METHODS: The study was conducted on primary cultures of hGCs. Gene expression was evaluated by RT-qPCR method. Statistical analysis was performed applying student t-test, as appropriate (P < 0.05) set for statistical significance. RESULTS: DCI is able to reduce the gene expression of CYP19A1, P450scc and insulin-like growth factor 1 receptor (IGF-1R) in dose-response manner. The presence of DCI impaired the increased expression of steroidogenic enzyme genes generated by the insulin treatment in gonadotrophin-stimulated hGCs. CONCLUSIONS: Insulin acts as co-gonadotrophin increasing the expression of steroidogenic enzymes genes in gonadotrophin-stimulated granulosa cells. DCI is an insulin-sensitizer that counteracts this action by reducing the expression of the genes CYP19A1, P450scc and IGF-1R. The ability of DCI to modulate in vitro ovarian activity of insulin could in part explain its beneficial effect when used as treatment for conditions associated to insulin resistance.

Clock drawing test: comparison between the Pfizer and the Shulman systems.

Cognitive decline can be screened by the clock drawing test (CDT), which has several versions. Objective: This survey aimed to analyze the correlation between two simple methods for scoring the CDT. Methods: This cross-sectional study was nested in the Elo-Creati cohort from Passo Fundo, Brazil and comprised 404 subjects. Two raters underwent previous training and scored the subjects' CDT according to both the Pfizer and Shulman systems. The inter-observer and intra-observer concordance within each method was analyzed with the Spearman's rank correlation coefficient, as well as the concordance of the scores between the two methods. Age and scholarity were also correlated with the scores. Results: Most of the participants were women (93.8%) and Caucasian (84.6%), with a mean age of 66.9 (±7.8) years and a scholarity of 10.9 years (±5.6). There was significant inter-observer (Pfizer: r=0.739, p£0.001; Shulman: r=0.727, p£0.001) and intra-observer correlation (Pfizer: rater 1, r=0.628, p≤0.001; rater 2, r=0.821, p≤0.001; Shulman: rater 1, r=0.843, p≤0.001; rater 2: r=0.819; p≤0.001). Intra-observer correlation was also observed comparing Pfizer and Shulman methods (rater 1: r=0.744; p≤0.001; rater 2: r=0.702; p≤0.001). There was weak correlation of the scores with scholarity (Pfizer: r=0.283, p£0.001; Shulman: r=0.244, p£0.001) and age (Pfizer: r=-0.174, p£0.001; Shulman: r=-0.170, p£0.001). More participants were classified with decreased cognition through the Pfizer system (rater 1: 44.3 vs. 26.5%; rater 2: 42.1 vs. 16.3%; p≤0.001). Conclusions: For this population, our results suggest that the Pfizer system of scoring CDT is more suitable for screening cognitive decline.

O déficit cognitivo pode ser triado pelo teste do desenho do relógio (TDR), que tem várias versões. Objetivo: Esta pesquisa visou avaliar a concordância entre dois métodos simples de TDR. Métodos: Estudo transversal, aninhado na coorte Elo-Creati de Passo Fundo, Brasil, que incluiu 404 sujeitos. Dois avaliadores previamente treinados analisaram o TDR dos participantes de acordo com os sistemas de Pfizer e de Shulman. A concordância inter e intraobservador foi analisada com o teste de coeficiente de correlação de postos de Spearman, assim como a concordância pela estatística kappa dos escores entre os métodos. Idade e escolaridade também foram correlacionados com os escores. Resultados: A maioria dos participantes era de mulheres (93,8%) e caucasianos (84,6%), com média de idade de 66,9±7,8 anos e de escolaridade de 10,9±5,6 anos. Houve significativa correlação interobservador (Pfizer: r=0,739, p£0,001; Shulman: r=0,727, p£0,001) e intraobservador (Pfizer: avaliador 1, r=0,628, p≤0,001; avaliador 2, r=0,821, p≤0,001; Shulman: avaliador 1, r=0,843, p≤0,001; avaliador 2: r=0,819; p≤0,001). Correlação intraobservador significativa também foi evidenciada comparando-se os sistemas de Pfizer e Shulman (avaliador 1: r=0,744; p≤0,001; avaliador 2: r=0,702; p≤0,001). Houve fraca correlação dos escores com escolaridade (Pfizer: r=0,283, p£0,001; Shulman: r=0,244, p£0,001) e idade (Pfizer: r=-0,174, p£0,001; Shulman: r=-0,170, p£0,001). Mais participantes foram classificados com declínio cognitivo com o sistema de Pfizer (avaliador 1: 44,3 vs. 26,5%; avaliador 2: 42,1 vs. 16,3%; p≤0,001). Conclusões: Nossos resultados sugerem que, para essa população, o sistema de Pfizer para avaliar o TDR é mais adequado para a triagem cognitiva.

Clock drawing test: comparison between the Pfizer and the Shulman systems / Teste do desenho do relógio: comparação dos sistemas de Pfizer e de Shulman

ABSTRACT Cognitive decline can be screened by the clock drawing test (CDT), which has several versions. Objective: This survey aimed to analyze the correlation between two simple methods for scoring the CDT. Methods: This cross-sectional study was nested in the Elo-Creati cohort from Passo Fundo, Brazil and comprised 404 subjects. Two raters underwent previous training and scored the subjects' CDT according to both the Pfizer and Shulman systems. The inter-observer and intra-observer concordance within each method was analyzed with the Spearman's rank correlation coefficient, as well as the concordance of the scores between the two methods. Age and scholarity were also correlated with the scores. Results: Most of the participants were women (93.8%) and Caucasian (84.6%), with a mean age of 66.9 (±7.8) years and a scholarity of 10.9 years (±5.6). There was significant inter-observer (Pfizer: r=0.739, p£0.001; Shulman: r=0.727, p£0.001) and intra-observer correlation (Pfizer: rater 1, r=0.628, p≤0.001; rater 2, r=0.821, p≤0.001; Shulman: rater 1, r=0.843, p≤0.001; rater 2: r=0.819; p≤0.001). Intra-observer correlation was also observed comparing Pfizer and Shulman methods (rater 1: r=0.744; p≤0.001; rater 2: r=0.702; p≤0.001). There was weak correlation of the scores with scholarity (Pfizer: r=0.283, p£0.001; Shulman: r=0.244, p£0.001) and age (Pfizer: r=-0.174, p£0.001; Shulman: r=-0.170, p£0.001). More participants were classified with decreased cognition through the Pfizer system (rater 1: 44.3 vs. 26.5%; rater 2: 42.1 vs. 16.3%; p≤0.001). Conclusions: For this population, our results suggest that the Pfizer system of scoring CDT is more suitable for screening cognitive decline.

RESUMO O déficit cognitivo pode ser triado pelo teste do desenho do relógio (TDR), que tem várias versões. Objetivo: Esta pesquisa visou avaliar a concordância entre dois métodos simples de TDR. Métodos: Estudo transversal, aninhado na coorte Elo-Creati de Passo Fundo, Brasil, que incluiu 404 sujeitos. Dois avaliadores previamente treinados analisaram o TDR dos participantes de acordo com os sistemas de Pfizer e de Shulman. A concordância inter e intraobservador foi analisada com o teste de coeficiente de correlação de postos de Spearman, assim como a concordância pela estatística kappa dos escores entre os métodos. Idade e escolaridade também foram correlacionados com os escores. Resultados: A maioria dos participantes era de mulheres (93,8%) e caucasianos (84,6%), com média de idade de 66,9±7,8 anos e de escolaridade de 10,9±5,6 anos. Houve significativa correlação interobservador (Pfizer: r=0,739, p£0,001; Shulman: r=0,727, p£0,001) e intraobservador (Pfizer: avaliador 1, r=0,628, p≤0,001; avaliador 2, r=0,821, p≤0,001; Shulman: avaliador 1, r=0,843, p≤0,001; avaliador 2: r=0,819; p≤0,001). Correlação intraobservador significativa também foi evidenciada comparando-se os sistemas de Pfizer e Shulman (avaliador 1: r=0,744; p≤0,001; avaliador 2: r=0,702; p≤0,001). Houve fraca correlação dos escores com escolaridade (Pfizer: r=0,283, p£0,001; Shulman: r=0,244, p£0,001) e idade (Pfizer: r=-0,174, p£0,001; Shulman: r=-0,170, p£0,001). Mais participantes foram classificados com declínio cognitivo com o sistema de Pfizer (avaliador 1: 44,3 vs. 26,5%; avaliador 2: 42,1 vs. 16,3%; p≤0,001). Conclusões: Nossos resultados sugerem que, para essa população, o sistema de Pfizer para avaliar o TDR é mais adequado para a triagem cognitiva.