ABSTRACT Purpose To determine enzymatic antioxidant and lipid peroxidation levels in seminal plasma of patients orchiectomized for testicular tumors. Materials and Methods The study included 52 patients: 26 control men and 26 orchiectomized patients for testicular tumor, of which 12 men had seminoma tumor and 14 men non-seminoma tumor. After semen analysis performed according to the WHO guidelines, an aliquot of semen was centrifuged and the seminal plasma was collected. Lipid peroxidation was performed by thiobarbituric acid reactive substances (TBARS) assay and antioxidant profile was assessed by analyzing catalase, glutathione peroxidase (GPx) and superoxide anion (SOD) activities using colorimetric assays with a standard spectrophotometer. Data were tested for normality and compared using one-way ANOVA (p<0.05). Results Seminoma and non-seminoma groups presented lower sperm concentration and morphology when compared to control group (p=0.0001). Both study groups (seminoma and non-seminoma) presented higher TBARS levels when compared to control group (p=0.0000013). No differences were observed for SOD (p=0.646) andGPx (p=0.328). It was not possible to access the enzymatic activity of catalase in any group. Conclusion Patients with testicular tumor present increased semen oxidative stress, but no differences were observed in antioxidant levels, even after orchiectomy. This indicates that most likely an increased generation of oxidative products takes place in these patients.

O objetivo deste estudo foi identificar a intenção de revelar ou não para o filho, familiares e amigos a ovodoação e os motivos subjacentes. Participaram do estudo dez casais receptores de oócitos do Programa de Ovodoação da Universidade Federal de São Paulo. Foram realizadas entrevistas individuais, e os dados foram analisados segundo o método qualitativo. A maioria dos participantes relatou que não havia contado sobre a ovodoação para outras pessoas, assim como não tinha a intenção de contar para o filho. Foram alegados diversos motivos, acompanhados de temores relacionados às possíveis reações das crianças. O sigilo sobre a doação de gametas é uma questão complexa, que não deve ser respondida com regras simples e gerais, devendo-se considerar as especificidades de cada família e o contexto em que vivem.

The aim of this study was to identify the intention among parents to disclose conception via egg donation to their child, relatives and friends and the underlying reasons. The study considered ten couples to have received oocytes via the Egg Donation Programme at the Federal University of São Paulo. Most couples preferred not to disclose their conception via egg donation to friends or relatives and did not intend to inform their child. Various reasons were reported, as well as fears over the future child's potential reaction. Disclosure regarding gamete donation is a complex issue and approaches should not apply simple and general rules, but rather take into consideration each family's specific circumstances and context.

A infertilidade interrompe um projeto de vida pessoal e do casal, produzindo sofrimento psíquico. O desenvolvimento das técnicas de reprodução assistida possibilitou a solução para casais que anteriormente não teriam alternativas de tratamento. Assim, a fertilização in vitro constitui uma nova fonte de esperança para ter um filho, mas, ao mesmo tempo, pode ser acompanhada de muitas dificuldades. O objetivo deste estudo foi comparar os estados emocionais de homens e mulheres submetidos à fertilização in vitro e verificar a relação entre estado emocional das mulheres e ocorrência de gravidez. Foram avaliados vinte casais submetidos ao primeiro ciclo de fertilização in vitro, entre o 5º e o 10º dia após a transferência dos pré-embriões. A avaliação foi realizada por meio de testes psicológicos e entrevistas. As mulheres apresentaram níveis mais altos de ansiedade e depressão que os homens. A autoestima delas mostrou-se mais baixa. Um quarto das mulheres que apresentaram sintomas psicoemocionais não engravidou, porém esta relação não foi significativa. Concluiu-se que as mulheres apresentaram mais ansiedade e sintomas depressivos que os homens e autoestima mais baixa. O grupo de mulheres com sintomas psicoemocionais apresentou tendência a não engravidar.

Infertility intrudes into the plans in one's personal life and the life of a couple, and causes mental suffering. In vitro fertilization is a new technique in reproductive technology and offers fresh hope of having children, but at the same time, it is considered to be a stressful experience. The aim of this study is to compare the emotional state of men and women undergoing in vitro fertilization and identify a possible relationship between women's emotional state and a successful outcome. Twenty couples in their first cycle of in vitro fertilization were included in this study. Men and women were submitted individually to psychological interviews and tests between 5 and 10 days after the transfer of pre-embryos. Women presented higher levels of anxiety and depression than men. Their self-esteem proved to be lower. Those women (25%) that presented psycho-emotional symptoms, did not get pregnant, however this relationship was not significant. Based on these findings, we conclude that women presented greater anxiety and symptoms of depression than the men and had lower self-esteem. The group of women with psycho-emotional symptoms displayed a tendency not to get pregnant.

O aumento da sobrevida dos pacientes que utilizam terapêutica antiretroviral altamente eficaz (HAART- Highly Active Antiretroviral Therapy) trouxe uma nova demanda de casais sorodiscordantes que desejam filhos. Como esses casais não podem abandonar o uso de preservativos, torna-se indispensável tratar o sêmen infectado com técnicas laboratoriais eficazes que além de isolar os melhores espermatozóides, reduzam a carga viral do HIV e HCV a níveis indetectáveis. Para isso, são utilizadas técnicas de semen washing, associadas a testes ultra sensíveis de biologia molecular. Após análise seminal, sêmen de 20 pacientes co-infectados HIV-HCV foram submetidos a fracionamento celular e isolamento de espermatozóides móveis através de método de densidade de gradiente descontínuo e swim-up. Posteriormente, testes para detecção do RNA do HIV e HCV foram aplicados nos sêmens totais e frações seminais obtidas. Em fase pré semen washing, o HIV foi detectado em 100% dos semens totais. Contrariamente, o HCV foi detectado em apenas uma amostra. Em fase pós semen washing, o HIV e HCV não foram detectados em nenhuma das frações seminais. A redução do HIV e do HCV através de semen washing mostra-se um método eficaz a indivíduos co-infectados HIV-HCV, apesar do encontro do HCV no sêmen ser raro.

INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

Every year there are 10 thousand new cases of patients victimized by spinal cord trauma (SCT) in the United States and it is estimated that there are 7 thousand new cases in Brazil. Eighty percent of patients are fertile males. Infertility in this patient group is due to 3 main factors resulting from spinal cord lesions: erectile dysfunction, ejaculatory disorder and low sperm counts. Erectile dysfunction has been successfully treated with oral and injectable medications, use of vacuum devices and penile prosthesis implants. The technological improvement in penile vibratory stimulation devices (PVS) and rectal probe electro-ejaculation (RPE) has made such procedures safer and accessible to patients with ejaculatory dysfunction. Despite the normal number of spermatozoa found in semen of spinal cord-injured patients, their motility is abnormal. This change does not seem to be related to changes in scrotal thermal regulation, frequency of ejaculation or duration of spinal cord damage but to factors related to the seminal plasma. Despite the poor seminal quality, increasingly more men with SCT have become fathers through techniques ranging from simple homologous insemination to sophisticated assisted reproduction techniques such as intracytoplasmic sperm injection (ICSI).

PURPOSE: Varicoceles are associated with impaired testicular function and male infertility, but the molecular mechanisms by which fertility is affected have not been satisfactorily explained. Spermatogenesis might be affected by increased scrotal temperature, such as that caused by varicocele. HSP90 is a molecular chaperone expressed in germ cells and is related to spermatogenesis, motility, and both heat and oxidative stress. Possible correlations between coding single region nucleotide polymorphisms (cSNPs) in the HSP90 gene in patients with varicocele associated with infertility were analyzed, and polymorphisms in these exons were characterized through DNA sequencing. MATERIALS AND METHODS: PCR-SSCP and DNA sequencing were used to search for mutations in 18 infertile patients with varicocele, 11 patients with idiopathic infertility and 12 fertile men. DNA was extracted from leucocytes for PCR amplification and SSCP analysis. DNA from samples with an altered band pattern in the SSCP was then sequenced to search for polymorphisms. RESULTS: Three silent polymorphisms that do not lead to amino acid substitutions were identified. CONCLUSION: Mutations in the HSP90 gene do not appear to be a common cause of male factor infertility. The low incidence of gene variation, or SNPs, in infertile men demonstrates that this gene is highly conserved and thus confirms its key role in spermatogenesis and response to heat stress.

OBJECTIVE: To search and to identify spermatozoa and spermatids, present in the ejaculate of non-obstructive azoospermic patients. MATERIALS AND METHODS: 27 patients, aged between 18 and 48 years, with initial diagnosis compatible with non-obstructive azoospermia, underwent up to 3 seminal samples, with assessment of macroscopic and microscopic parameters differentiated for each sample. In the first sample, 5 µL of semen were analyzed in a Horwell chamber in order to assess the presence or absence of spermatozoa. The procedure was repeated with 2 other aliquots. In the absence of spermatozoa, the entire sample was transferred to a conic tube and following centrifugation the sediment was freshly analyzed. The second seminal sample was collected only when no spermatozoa were found in the first sample and the research was performed in the same way. In cases where spermatozoa were not seen, the sample was centrifuged and the obtained sediment was stained by the panoptic method and observed under common light microscopy (1250X). The third seminal sample was collected only in cases when patients had not shown spermatozoa in the first and second seminal samples. RESULTS: 4/27 (14.8%) patients presented spermatozoa in the first seminal sample and 6/23 (26.1%), in the second seminal sample. No spermatozoa were seen in the third sample, however, 11/17 (64.7%) presented spermatids. CONCLUSION: In clinical situations where the initial diagnosis is non-obstructive azoospermia, one single routine seminal analysis is not enough to confirm this diagnosis and the analysis of the centrifuged sediment can have relevant clinical consequences. Among patients considered non-obstructive azoospermic, when duly assessed, 37% presented spermatozoa and 64.7%, spermatids.

OBJECTIVE: Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. MATERIALS AND METHODS: Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v) with the cryoprotector containing glycerol (Test yolk buffer). One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI), while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III) (group II - GII). The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog%) and percentage of live spermatozoa (Vit%). After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h). The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. RESULTS: There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p < 0.001). Values of Prog% and Vit% were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog% among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006). After cryoprotector removal, Prog% has been kept unaltered, in both groups, during the first 3 hours of incubation, although being superior in group I (p = 0,04). There was a significant decrease in Prog% after 24 hours of incubation, in both groups (p < 0,01). CONCLUSION: For human semen samples with low initial quality, freezing through vapor technique or through the automated technique showed to be equivalent in regarding recovery of live spermatozoa with progressive motility. The effects of dilution and centrifugation to remove the cryoprotector had a negative impact only in samples frozen through the automated technique. In both techniques, progressive motility is kept constant during the first 3 hours after thawing and removal of the cryoprotector, but is drastically diminished by the end of an incubation period of 24 hours.

O artigo apresenta dados preliminares de um estudo prospectivo com uma amostra de alunos saudáveis variando de idade entre 14 e 17 anos, de uma escola técnica em São Paulo. Foram avaliados 96 adolescentes estágio Tanner 5 e 31 estágio Tanner 4: o volume testicular foi medido utilizando um orquidômetro de Prader, e o espermograma foi realizado de acordo com os procedimentos padronizados. O volume testicular mediano, tanto direito quanto esquerdo, era 20,0ml nos alunos Tanner 5 e 15,0ml nos alunos Tanner 4. Não foram constatadas diferenças significativas no volume ou motilidade espermática. A concentração mediana era 66,0 milhões/ml para Tanner 5 e 47,0 milhões/ml para Tanner 4. Os padrões morfológicos evidenciaram formas anormais em 81,9% dos adolescentes Tanner 5 e 93,6% dos Tanner 4. A oligospermia (concentração espermática < 5 milhões/ml) foi observada em 7,3% dos indivíduos Tanner 5 e 12,9% dos Tanner 4. A azoospermia foi observada em 3 alunos (1,8%), além de uma contagem abaixo de 1,0 em 8 alunos (4,8%). Os autores discutem os resultados observados, analisando as implicações para o desenvolvimento biológico a partir de possíveis exposições ambientais.

The article presents preliminary data from a prospective investigation in a sample of healthy 14-17-year-old students from a technical school in Sao Paulo, Brazil. Ninety-six Tanner stage 5 and thirty-one Tanner stage 4 adolescents were evaluated: testicular volume was measured using a Prader orchidometer, and semen analysis was performed according to standard procedures. Median testicular volume was 20.0ml among Tanner 5 students and 15.0ml in Tanner 4 students in both the right and left testes. No significant differences were found in sperm volume or motility. Median concentration was 66.0 million/ml for Tanner 5 and 47.0 million/ml for Tanner 4 subjects. Morphological patterns showed abnormal forms in 81.9% of Tanner 5 and 93.6% of Tanner 4 adolescents. Oligospermia (sperm concentration < 5 million/ml) was observed in 7.3% of Tanner stage 5 and 12.9% of Tanner stage 4 individuals. Azoospermia was observed in 3 students (1.8%), with counts less than 1.0 in 8 students (4.8%). The authors discuss the observed results, analyzing the potential implications arising from biological development and potential environmental exposures.

O artigo apresenta dados preliminares de um estudo prospectivo com uma amostra de alunos saudáveis variando de idade entre 14 e 17 anos, de uma escola técnica em São Paulo. Foram avaliados 96 adolescentes estágio Tanner 5 e 31 estágio Tanner 4: o volume testicular foi medido utilizando um orquidômetro de Prader, e o espermograma foi realizado de acordo com os procedimentos padronizados. O volume testicular mediano, tanto direito quanto esquerdo, era 20,0ml nos alunos Tanner 5 e 15,0ml nos alunos Tanner 4. Não foram constatadas diferenças significativas no volume ou motilidade espermática. A concentração mediana era 66,0 milhões/ml para Tanner 5 e 47,0 milhões/ml para Tanner 4. Os padrões morfológicos evidenciaram formas anormais em 81,9% dos adolescentes Tanner 5 e 93,6% dos Tanner 4. A oligospermia (concentração espermática < 5 milhões/ml) foi observada em 7,3% dos indivíduos Tanner 5 e 12,9% dos Tanner 4. A azoospermia foi observada em 3 alunos (1,8%), além de uma contagem abaixo de 1,0 em 8 alunos (4,8%). Os autores discutem os resultados observados, analisando as implicações para o desenvolvimento biológico a partir de possíveis exposições ambientais.

The article presents preliminary data from a prospective investigation in a sample of healthy 14-17-year-old students from a technical school in Sao Paulo, Brazil. Ninety-six Tanner stage 5 and thirty-one Tanner stage 4 adolescents were evaluated: testicular volume was measured using a Prader orchidometer, and semen analysis was performed according to standard procedures. Median testicular volume was 20.0ml among Tanner 5 students and 15.0ml in Tanner 4 students in both the right and left testes. No significant differences were found in sperm volume or motility. Median concentration was 66.0 million/ml for Tanner 5 and 47.0 million/ml for Tanner 4 subjects. Morphological patterns showed abnormal forms in 81.9% of Tanner 5 and 93.6% of Tanner 4 adolescents. Oligospermia (sperm concentration < 5 million/ml) was observed in 7.3% of Tanner stage 5 and 12.9% of Tanner stage 4 individuals. Azoospermia was observed in 3 students (1.8%), with counts less than 1.0 in 8 students (4.8%). The authors discuss the observed results, analyzing the potential implications arising from biological development and potential environmental exposures.

CONTEXTO: A verificação da maturidade endometrial, elemento diagnóstico necessário na avaliação do casal com queixa de infertilidade, vem sendo feita desde 1950 através do critério de datação histológica de Noyes. No entanto, não existe um consenso em relação ao período da fase lútea mais adequado para a colheita. OBJETIVO: Avaliar a correlação entre as datações histológicas de duas amostras de endométrio colhidas nos dias 6 e 10 da fase lútea de um mesmo ciclo menstrual. LOCAL: Setor de Reprodução Humana da Universidade Federal de São Paulo. TIPO DE ESTUDO: Estudo prospectivo. Constou da comparação entre duas datações de endométrio num mesmo ciclo menstrual. PARTICIPANTES: 25 pacientes com queixa de infertilidade tiveram um ciclo menstrual monitorizado por ultra-sonografia e medida plasmática de LH, para demonstração de ovulação. PROCEDIMENTO: Biópsias de endométrio foram feitas nos dias LH+6 e LH+10 da fase lútea, considerando-se o dia seguinte ao do pico de LH como LH+1. A datação foi feita de acordo com critério morfométrico, considerando-se o endométrio como fora de fase, se o atraso de maturação mínimo fosse de um dia. No dia LH+6 foi feita dosagem de progesterona plasmática. RESULTADOS: Todas as pacientes apresentaram ciclos ovulatórios (média dos valores de pico de LH: 47,3 U/L; média dos diâmetros foliculares no dia do pico de LH: 18,9 mm; média das espessuras do endométrio no dia do pico de LH: 10,3 mm; média das concentrações de progesterona plasmática no dia LH+6: 14,4 ng/ml.). Em 14 pacientes, as duas biópsias estavam em fase. Houve atraso de maturação apenas no dia LH+6 em cinco pacientes; apenas no dia LH+10 em três pacientes e, nos dois dias, em três pacientes. Não houve diferença estatística entre esses valores (teste de McNemar, p=33,36%). CONCLUSÕES: Os resultados sugerem que a colheita do endométrio em qualquer dos dias (sexto ou décimo) da fase lútea fornece resultados semelhantes em relação à maturidade endometrial.

CONTEXT: Endometrial maturation, important in the diagnosis of infertile couples, has been evaluated since 1950 using the Noyes criteria. Nevertheless, there is no consensus regarding the most suitable period of the luteal phase for performing the biopsy. OBJETIVE: This study evaluated the correlation between the histological dating of two endometrial biopsies performed in the same menstrual cycle, on luteal phase days six and ten. DESIGN: Prospective study. SETTING: Human Reproduction Division of the Federal University of São Paulo, referral center. PATIENTS:Twenty-five women complaining of infertility had their menstrual cycles monitored by ultrasound and LH plasma levels, to obtain evidence of ovulation. PROCEDURES: Endometrial biopsies were performed on luteal phase days LH+6 and LH+10 (luteal phase day 1 = LH+1 = the day that follows LH peak). Dating was done according to morphometric criteria, in which an endometrium sample is considered out of phase if the minimum maturation delay is one day. On day LH+6, blood was drawn for plasma progesterone level determination. RESULTS: All patients had an ovulatory cycle (mean LH peak: 47.4 U/L; mean follicular diameter on LH peak day: 18.9 mm; mean endometrial thickness on LH peak day: 10.3 mm; mean plasma progesterone level on day LH+6: 14.4 ng/ml). 14 patients had both biopsies in phase; 5 patients had out of phase biopsies only on day LH+6; 3 had out of phase biopsies only on day LH+10 and 3 patients had out of phase biopsies on both days. McNemar's test showed no statistical difference between these data (p>33.36%). CONCLUSIONS: The correlation found between the endometrial datings suggests that biopsies performed on either of these two days are suitable for evaluation of endometrial maturation.