Claudia Cristina Paro de Paz

Differentially expressed genes in eutopic and ectopic endometrium of women with endometriosis

OBJECTIVE To elucidate the potential mechanisms involved in the physiopathology of endometriosis. We analyzed the differential gene expression profiles of eutopic and ectopic tissues from women with endometriosis. DESIGN Prospective laboratory study. SETTING University hospital. PATIENT(S) Seventeen patients in whom endometriosis was diagnosed and 11 healthy fertile women. INTERVENTION(S) Endometrial biopsy specimens from the endometrium of healthy women without endometriosis and from the eutopic and ectopic endometrium tissues of patients with endometriosis were obtained in the early proliferative phase of the menstrual cycle. MAIN OUTCOME MEASURE(S) Six paired samples of eutopic and ectopic tissue were analyzed by subtractive hybridization. To evaluate the expression of genes found by rapid subtraction hybridization methods, we measured CTGF, SPARC, MYC, MMP, and IGFBP1 genes by real-time polymerase chain reaction in all samples. RESULT(S) This study identified 291 deregulated genes in the endometriotic lesions. Significant expression differences were obtained for SPARC, MYC, and IGFBP1 in the peritoneal lesions and for MMP3 in the ovarian endometriomas. Additionally, significant differences were obtained for SPARC and IGFBP1 between the peritoneal and ovarian lesions. No significant differences were found for the studied genes between the control and the eutopic endometrium. CONCLUSION(S) This study identified 291 genes with differential expression in endometriotic lesions. The deregulation of the SPARC, MYC, MMP3, and IGFBPI genes may be responsible for the loss of cellular homeostasis in endometriotic lesions.

Resposta do híbrido de sorgo-sudão à adubação nitrogenada e potássica: composição química e digestibilidade in vitro da matéria orgânica

A pesquisa foi instalada no Setor de Forragicultura da FCAV/UNESP-Jaboticabal, objetivando avaliar a composicao quimica e a digestibilidade in vitro da materia orgânica (DIVMO) do hibrido de Sorgo-sudao cv. AG 2501C, no outono e inverno. O manejo da pastagem foi conduzido simulando o sistema de lotacao intermitente. O experimento foi desenvolvido de marco a setembro de 2002. A forrageira foi submetida a nove tratamentos: tres niveis de nitrogenio (100, 200 e 300 kg de N/ha) e tres niveis de potassio (0, 80 e 160 kg de K2O/ha), em delineamento experimental em blocos casualizados e parcelas subdivididas. A adubacao nitrogenada e potassica nao foram significativas para a DIVMO. O nitrogenio influenciou a proteina bruta (PB) com valores de 15,1; 16,4 e 15,7 %, a fibra em detergente neutro (FDN) com valores de 65,3; 65,8 e 64,5% e fibra em detergente acido (FDA) com 35,5; 37,8 e 39,6% para 100; 200 e 300 kg N/ha. O potassio aumentou significativamente a lignina das plantas. O melhor nivel obtido foi 100 kg/ha de nitrogenio, sem potassio.

Influência de fatores de ambiente sobre a contagem de células somáticas e sua relação com perdas na produção de leite de vacas da raça Holandesa

Cumulative milk yield at 305 days (MY-305) and somatic cell count (SCC) from the first to the fifth lactation of Holstein cows were analyzed in this study. Somatic cell count was logarithmically transformed to somatic cell score (SCS) that was then included in the model not only as a dependent but also as an independent variable. Year, season, and parity order all significantly affected the average SCS in this study. Data of the effect of SCS on milk yield showed significant losses on MY-305 because of the observed negative linear relationship between these two variables. Somatic cell score affected differently MY-305 from different parity orders with greater effect after the 3rd parity. Although clinical mastitis seems to cause greater economic losses, prevention and control of sub-clinical mastitis deserve more attention due to its greater prevalence leading to significant reduction on herd productivity.Cumulative milk yield at 305 days (MY-305) and somatic cell count (SCC) from the first to the fifth lactation of Holstein cows were analyzed in this study. Somatic cell count was logarithmically transformed to somatic cell score (SCS) that was then included in the model not only as a dependent but also as an independent variable. Year, season, and parity order all significantly affected the average SCS in this study. Data of the effect of SCS on milk yield showed significant losses on MY-305 because of the observed negative linear relationship between these two variables. Somatic cell score affected differently MY-305 from different parity orders with greater effect after the 3rd parity. Although clinical mastitis seems to cause greater economic losses, prevention and control of sub-clinical mastitis deserve more attention due to its greater prevalence leading to significant reduction on herd productivity.

Glycodelin expression in the endometrium of healthy women and in the eutopic and ectopic endometrium of women with endometriosis.

OBJECTIVE To analyze the expression of the glycodelin gene to better understand the molecular environment of endometriotic lesions and to elucidate the potential mechanisms that underlie the complex physiopathology of endometriosis. DESIGN Prospective laboratory study. SETTING University hospital. PATIENT(S) Eleven healthy fertile women and 17 patients with endometriosis in the early proliferative phase of the menstrual cycle. INTERVENTION(S) Endometrial biopsy specimens were obtained from the endometrium of healthy women without endometriosis (controls) and from eutopic and ectopic endometrium tissues (pelvic and ovarian endometriotic implants) of endometriosis patients. MAIN OUTCOME MEASURE(S) The glycodelin relative expression level by real-time polymerase chain reaction (PCR) analysis. RESULT(S) The glycodelin down-regulation found in the endometriotic lesions was 332.26 and 123.17-fold lower, respectively, when compared with the eutopic tissue and the control endometrium. CONCLUSION(S) Glycodelin may be one of the molecules that contributes to the loss of cellular homeostasis in endometriotic lesions.

Increased Circulating MMP-2 Levels in Infertile Patients With Moderate and Severe Pelvic Endometriosis

The current study compares the levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the follicular fluid (FF) of infertile patients with and without endometriosis submitted to ovarian stimulation for in vitro fertilization and the levels of MMP-2 in the serum of the same patients. We also evaluated whether the severity of endometriosis can influence serum and/or FF concentration of these metalloproteinases. A cross-sectional study was conducted on 30 patients: stage I/II endometriosis (n = 10), stage III/IV endometriosis (n = 10), and control (infertility due to tubal and/or male factor; n = 10). Blood samples for the analysis of MMP-2 levels were obtained during the early follicular phase of the menstrual cycle. The FF samples for the analysis of MMP-2 and MMP-9 were obtained on the day of oocyte retrieval. The concentrations of MMP-2 and MMP-9 were determined by zymography. No intragroup or intergroup difference was observed in MMP-2 or MMP-9 levels in FF. Significantly higher MMP-2 levels were detected in the serum of infertile women with stage III/IV endometriosis compared to women with stage I/II endometriosis. In conclusion, no differences were observed in the follicular levels of MMP-2 and MMP-9 between infertile patients with and without endometriosis. However, the levels of MMP-2 were significantly higher in the serum of infertile women with advanced stages of endometriosis. Taken together, the present results demonstrate that advanced pelvic endometriosis severity is related to higher serum MMP-2 levels but does not influence follicular MMP-2 or MMP-9 levels in periovulatory follicles obtained from stimulated cycles.

Down-regulation of the CYP19A1 gene in cumulus cells of infertile women with endometriosis

Aromatase plays a fundamental role in the establishment of oocyte quality, which might be compromised in infertile women with endometriosis. The expression of the CYP19A1 gene (that encodes aromatase) was compared in cumulus cells and oestradiol concentrations in the follicular fluid of infertile women with and without endometriosis submitted to ovarian stimulation for intracytoplasmic sperm injection. Cumulus cells were isolated and the expression of the CYP19A1 was quantitated through real-time polymerase chain reaction. Oestradiol concentrations in follicular fluid were measured by chemiluminescence immunoassay. A lower expression of the CYP19A1 in the cumulus cells of infertile women with endometriosis was observed compared with controls (0.17 ± 0.13 and 0.56 ± 0.12, respectively), and no significant difference in the follicular fluid oestradiol concentrations was observed between groups. Our results show reduced expression of the CYP19A1 in cumulus cells of infertile women with endometriosis, which may play a role in the pathogenesis of endometriosis-related infertility.

IGF-I and IgG serum concentrations and in mammary secretions in cows treated with rbST prepartum

Foram utilizadas 42 vacas holandesas gestantes e multiparas, distribuidas ao acaso em dois grupos, com o objetivo de se determinar possiveis alteracoes na concentracao de IGF-I no colostro e nas secrecoes lacteas em resposta a mudancas na concentracao serica de IGF-I pre-parto, alem de se avaliar comparativamente a flutuacao serica pre-parto de IGF-I em relacao a IgG. O grupo tratado (n=21) recebeu 500 mg de somatotropina bovina recombinante (rbST) e o grupo controle (n=21) injecoes de vitamina E. Os tratamentos foram iniciados 35 dias pre-parto e repetidos a cada 14 dias ate o parto. Semanalmente, ate o parto, avaliou-se o escore corporal (EC) e coletaram-se amostras de sangue da veia mamaria. Foram amostrados o colostro e as secrecoes lacteas diariamente por sete dias pos-parto. O EC e a concentracao serica de acidos graxos nao-esterificados nao diferiram entre os grupos. O grupo rbST apresentou maiores concentracoes sericas de IGF-I na segunda e quarta semanas apos o inicio dos tratamentos, nao sendo encontradas diferencas entre os tratamentos ao parto. A concentracao de IGF-I foi superior no colostro das vacas tratadas com rbST, nao diferindo nas secrecoes subsequentes. As concentracoes sericas de IgG nao diferiram entre tratamentos durante todo o periodo experimental, assim como as do colostro e demais secrecoes lacteas. As concentracoes sericas de IGF-I e IgG variaram em funcao do periodo experimental, diferindo na amplitude e refletindo as grandes mudancas que ocorrem no ultimo mes de gestacao.

Higher SOD1 Gene Expression in Cumulus Cells From Infertile Women With Moderate and Severe Endometriosis

It is questioned whether worsening of oocyte quality and oxidative stress (OS) are involved in the pathogenesis of infertility related to endometriosis and in compromised intracytoplasmic sperm injection (ICSI) outcomes. Cumulus cells (CCs) protect oocytes from entering apoptosis induced by OS. Thus, we carried out a case–control study comparing expression of superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), and glutathione peroxidase 4 (GPX4; genes encoding for the main antioxidant enzymes) in CCs from mature oocytes of 26 infertile patients with minimal/mild endometriosis, 14 patients with moderate/severe endometriosis, and 41 controls undergoing controlled ovarian stimulation for ICSI, using real-time polymerase chain reaction. As a secondary objective, we investigated the interaction between the expression of these genes and clinical pregnancy (CP) by a statistical model. Only infertile women with moderate/severe endometriosis showed increased expression of the SOD1 in CCs compared to women with minimal/mild endometriosis and controls, with a positive interaction between increased expression and the occurrence of CP, suggesting that SOD1 might be a potential biomarker of CP following ICSI.

Ajuste de modelos não-lineares em estudos de associação entre polimorfismos genéticos e crescimento em bovinos de corte

Weights at birth, at weaning and monthly from 8 to 19 months of age of the 11 genotype identified by combination of polymorphic RFLPs of the genes kappa-casein-HinfI (CSN3): AA and AB, growth hormone-AluI (GH): LL and LV, and b-lactoglobulin-HaeIII (LGB): AA, AB and BB (G1=AALLAA, G2=AALLAB, G3=AALLBB, G4=AALVAB, G5=AALVBB, G6=ABLLAA, G7=ABLLAB, G8=ABLLBB, G9=ABLVAA, G10=ABLVAB e G11=ABLVBB) were used. Data were obtained from three genetic groups: ½Canchim-Nelore (CN), ½Angus-Nelore (AN) and ½Simmental-Nelore (SN), born in 1998 and 1999 in Southeast Brazil. From five nonlinear models studied: Brody, Von Bertalanffy, Richards, Gompertz and Logistic, the more adequate was the last one. The parameters A (asymptotic value), k (maturing rate) and m (inflection point) estimated by Logistic model for each animal, were analyzed by ordinary least squares, by a linear model that included, besides the overall mean, the fixed effects of the genotype, year of birth, sex and feed management. For the CN genetic groups, was detected significant effect of genotype for A and k parameters estimates. The genotypes G3 showed inferior value of A and superior of k in relation to G7 and G8. Considering the AN and SN genetic groups, the effects of genotype on A, k and m parameters were not significant. Nonlinear model application in this study of association between genetic marker and weight trait in beef cattle provided a fitted analysis of the shape of the growth curve regarding the genotype for the CSN3, GH and LGB genes.

Influência de polimorfismos genéticos sobre os parâmetros da curva de crescimento em bovinos de corte

ABSTRACT - Records of the weights at birth, weaning and monthly from 8 to 19 months of age from three genetic groups: ½Canchim-Nellore (CN), ½Angus-Nellore (AN) e ½Simmental-Nellore (SN), born in 1998 and 1999 in Southeast Brazil, were used to fit a Logi sticnonlinear model that included the fixed effects of the contemporary group and genotype of the genes kappa-casein- Hinf I (CSN3): AAand AB, growth hormone- Alu I (GH): LL and LV, and β-lactoglobulin- Hae III (LGB): AA, AB and BB, to examine the effect of thesemarkers on the growth curve. The Logistic model used to explain the growth of the CN, AN and SN genetic groups, was influenced bythe genotypes of the CSN3, GH e LGB markers. The major differences started at 12-13 months of age. The value of the maturing ra te( k ) of the AA genotype for CSN3 was superior in relation to AB genotype in the CN, AN and SN genetic groups. However, there wasobserved small difference in estimate of the asymptotic value ( A ) for the AA and AB genotypes in CN and SN genetic groups. For theGH polymorphism there were inferior values of the parameters