Willingness of Infertile Couples to Pay for In Vitro Fertilization Treatment in the Integrated Human Reproduction Section of the Escola Paulista de Medicina, São Paulo Federal University.

OBJECTIVE: To describe the willingness to pay (WTP) of infertile couples for in vitro fertilization (IVF) treatment. METHOD: This was a prospective study with an anonymous questionnaire for infertile couples in an academic setting. Clinical characteristics were analyzed by a Student's t test or Mann-Whitney test, categorical variables were compared by a chi-square or Fisher exact test, and correlations were assessed using a Spearman's test. An alpha of 5% was adopted. RESULTS: Mean female and male ages were 31.5 and 35.9 years, respectively; 80.2% were married; 19.8% were in consensual union; 48.1% of women had college degrees; and 49.4% of men had a high school education. Most women (77.8%) and men (75.3%) were white, with a household income of class C. Average duration of union was 8.5 years, and average infertility was 4.7 years. Using a willingness-to-pay (WTP) evaluation and the technique of "direct questioning," the average value was determined to be R$18 720.18 (by payment scale R$22 831.17). WTP positively correlated with household income and each woman's education level. Previous parenthood or use of public health system negatively correlated with WTP. CONCLUSION: We conclude that the higher the couple's monthly income and the woman's educational level, the higher the WTP for an IVF treatment; previous parenthood determined a lower WTP for an IVF treatment, and previous use of the Brazilian Unified Health System, determined a lower WTP for an IVF treatment.

Expression of the Fas-ligand gene in ejaculated sperm from adolescents with and without varicocele.

PURPOSE: To assess FasL mRNA levels in ejaculated sperm from adolescent patients with and without varicocele. METHODS: Semen was obtained by masturbation following 2-4 days of ejaculatory abstinence, from 14 adolescents with varicocele grades II and III (study group), and 20 adolescents without varicocele (control group). Seminal analysis was done according to World Health Organization guidelines and morphology using Kruger's strict criteria. The Fas-ligand (FasL) gene expression was performed using reverse transcription and real-time quantitative polymerase chain reaction (RQ-PCR) analysis, according to the expression level of the housekeeping cyclophilin A gene. A Student's t-test was applied to compare the groups, and Spearman's rank test in order to verify possible correlations (p < 0.05). RESULTS: Quantitative RQ-PCR demonstrated that the expression of FasL mRNA in sperm from the varicocele group was higher than in the control group. Also, sperm concentration was higher in the controls, when compared to the varicocele group. When submitted to correlation analysis, adolescents with varicocele presented a correlation between sperm concentration and FasL gene expression levels (r = -0.470), not observed in controls. CONCLUSION: Our results allow us to conclude that, in adolescents with varicocele presenting lower sperm concentration, FasL mRNA levels are higher than in adolescents without varicocele.

Expression of the HSPA2 gene in ejaculated spermatozoa from adolescents with and without varicocele.

OBJECTIVE: To evaluate mRNA expression of the HSPA2 gene in ejaculated spermatozoa in adolescents with and without varicocele. DESIGN: Controlled prospective study. SETTING: Patients in an academic research environment. PATIENTS: Adolescent patients with clinical diagnosed bilateral varicocele grades II and III, and adolescent patients without varicocele. INTERVENTION: Reverse-transcription-polymerase chain reaction (RT-PCR) analysis of HSPA2 gene expression in adolescents with and without varicocele. MAIN OUTCOME MEASURE: Comparative RT-PCR expression analysis of HSPA2 gene mRNA, compared to the housekeeping beta-actin gene. RESULTS: Sperm from adolescents with varicocele and oligozoospermia had significantly lower levels of HSPA2 gene expression than both adolescents without varicocele (controls) and adolescents with varicocele and normal sperm concentration (P<.05). The latter group had a nonsignificant increase in gene expression compared to the control group. CONCLUSIONS: This is the first report on HSPA2 gene expression in ejaculated spermatozoa from adolescents and its relationship with varicocele pathology. Results demonstrated that HSPA2 expression was down-regulated in adolescents with varicocele and oligozoospermia compared to controls. There was a higher, albeit nonsignificant, gene expression in adolescents with varicocele and normal sperm concentration than in controls. We speculate that expression levels of this gene might be used as a molecular marker for the acquisition of thermal tolerance in ejaculated spermatozoa.

Sperm nuclear DNA fragmentation in adolescents with varicocele.

OBJECTIVE: To verify if sperm from adolescents with varicocele have an increased rate of DNA fragmentation when compared with adolescents without varicocele. DESIGN: Controlled prospective study. SETTING: Patients in an academic research environment. PATIENT(S): Adolescent patients with a clinical diagnosed bilateral varicocele, grades II and III, and adolescent patients without a varicocele. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Rate of sperm DNA fragmentation as assessed by the Comet assay, graded as class I (no DNA fragmentation), II (little DNA fragmentation), III (meaningful DNA fragmentation), or IV (high DNA fragmentation). RESULT(S): A higher percentage of cells with no DNA fragmentation (class I) was found in the nonvaricocele group (47.62 +/- 7.69) when compared with the varicocele group (27.52 +/- 10.73). A higher percentage of sperm with class III and class IV DNA fragmentation was found in the varicocele group (20.43 +/- 8.97, and 19.57 +/- 10.68) when compared with the nonvaricocele group (11.38 +/- 5.55, and 5.71 +/- 2.35). CONCLUSION(S): Although standard semen analysis showed no difference between the groups, adolescents with varicocele have an increase in sperm nuclear DNA fragmentation. Thus, DNA fragmentation evaluation could be important in deciding treatment options for adolescent patients.

Reversed-phase high-performance liquid chromatography separation of adrenal steroids prior to radioimmunoassay: application in congenital adrenal hyperplasia.

21-Hydroxylase deficiency (21-OHD) is the most common form of congenital adrenal hyperplasia (CAH), followed by 11beta-hydroxylase deficiency (11beta-OHD). Diagnostic serum markers for these conditions are 17-hydroxyprogesterone (17-OHP) and 11-desoxycortisol (S), respectively. In 21-OHD, the large amounts of 17-OHP are further 11beta-hydroxylated to form 21-deoxycortisol (21-DF), making it also an excellent marker of this disease. These steroids can be measured in blood by radioimmunoassay (RIA). In this paper, we report the use of high-performance liquid chromatography (HPLC) for steroid purification, prior to RIA determinations of 21-DF, S, 17-OHP, and testosterone (T) in ether-extracted serum. The chromatographic separation is developed in a BDS-Hypersil column using water-methanol (53:47, v/v) as the mobile phase. The method is applied to 35 patients with the classic form of 21-OHD (18 females, 17 males, 5.1-14.2 years old) and 2 with 11beta-OHD (1 female, 1 male, 9.5 and 12.6 years old). Thirteen control children (5 females and 8 males, 5.2-15.2 years) are also studied. The results obtained for all measured steroids are compatible with those reported in the literature. The method is precise, and recovery is adequate. The HPLC technique proved to be of value for the purification of several steroids from single serum samples prior to RIA in patients with CAH.

Aplicaçäo da cromatografia líquida de alta eficiência na determinaçäo de estrôgenios eqüinos / The use of high-performance liquid chromatography for the determination of equine estrogens

Os estrogênios eqüinos, amplamente empregados na terapia de reposição hormonal em mulheres menopausadas, são encontrados em formulações de concentrações pré-estabelecidas disponíveis comercialmente. No presente trabalho, 17`alfaï-diidroeqüilina (A), eqüilina (Eq) e estrona (E1) foram determinadas empregando cromatografia líquida de alta eficiência (CLAE) com separação por fase reversa e detecção por ultravioleta em 220, 230 240 e 280 nm. Utilizou-se coluna cromatográfica ODS HYPERSIL©, fase móvel composta de água-acetronitrila (60:40 v/v) com fluxo de 1,0 mL/min. A testosterona foi utilizada como padrão de referência (tR = 4,6 min)...

Resposta esteroidogênica induzida por hCG em garanhöes jovens Mangalarga: testosterona e sulfato de estrona plasmáticos / HCG induced steroidogenic response in Mangalarga young stallions: plasma testosterone and estrone sulphate

Avaliou-se a resposta esteroidogênica aguda da célula de Leydig em sete garanhöes jovens da raça Mangalarga (2,6 a 4,5 anos), através de estimulaçäo com gonadotrofina coriônica humana (hCG). Os animais foram tratados com 5.000 UI (hCG) Vetecorr, i.v., in bollus, às 8 horas. Amostras de plasma foram obtidas por venopuntura às 7 horas (basal), 4, 24, 48, 72 horas pós-estímulo (28 a 31 de agosto). Testosterona (T) e sulfato de estrona (E1SO4) foram dosados por radioimunoensaio (RIE) c.v. £ 5 por cento. Os dados foram analisados pelos testes Friedman e Wilcoxon Matched-Pairs Signed Ranks (p £ 0,05). A magnitude da produçäo hormonal após o estímulo foi calculada pela Variaçäo Percentual (D por cento). As 48 horas do teste, 57,1 por cento dos animais (n = 4) apresentaram pico de T com um aumento de 5,7 vezes com relaçäo ao basal. Embora em 57,1 por cento (n = 4) tenha ocorrido a resposta máxima de E1SO4 às 4 horas, o valor máximo foi de 1,6 vez às 24 horas. A hCG pode ser empregada para estimular a resposta esteroidogênica da célula de Leydig e a dose de 5.000 UI é suficiente para estimular a esteroidogênese testicular. Em condiçöes basais, as concentraçöes plasmáticas de estrógenos säo superiores às dos andrógenos. Este protocolo pode ser usado para identificar problemas de origem endócrina em cavalos desta raça