Antioxidant enzyme profile and lipid peroxidation products in semen samples of testicular germ cell tumor patients submitted to orchiectomy

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.

Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing / Detecção do RNA do HIV e HCV em sêmen de homens brasileiros, usando PCR multiplex antes e depois do "semen washing"

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 percent 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.

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 por cento 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.

Search and identification of spermatozoa and spermatids in the ejaculate of non-obstructive azoospermic patients

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 æL 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 percent) patients presented spermatozoa in the first seminal sample and 6/23 (26.1 percent), in the second seminal sample. No spermatozoa were seen in the third sample, however, 11/17 (64.7 percent) 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 percent presented spermatozoa and 64.7 percent, spermatids.

Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men

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 percent) and percentage of live spermatozoa (Vit percent). After defrosting, Prog percent 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 percent and Vit percent from before freezing to after defrosting in both groups, I and II (p < 0.001). Values of Prog percent and Vit percent were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog percent among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006). After cryoprotector removal, Prog percent 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 percent 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...

Propriedades do muco cervical no período periovulatório: variação da elasticidade e da cristalização com a concentração plasmática de estradiol / Cervical mucus properties in the periovulatory period: spinability and ferning variations as related to plasma estradiol levels

O objetivo deste estudo é verificar a variação dos escores de cristalização e elasticidade do muco cervical com a concentração plasmática de estradiol, e sua relação com o período periovulatório. Trata-se de estudo prospectivo em pacientes com queixa de infertilidade, que tiveram um ciclo menstrual monitorizado por ultra-sonografia e dosagens hormonais de estradiol. Noventa e uma pacientes com queixa de infer-tilidade conjugal e ciclos menstruais regulares, com sinais ultra-sonográficos de ovulação e sem doença cervical participaram do estudo, no Setor de Reprodução Humana da UNIFEP/EPM. Diariamente, a partir do nono dia do ciclo e até a ovulação, foi dosado estradiol plasmático, foram verificadas a cristalização e a elasticidade do muco cervical e realizadas ultra-sonografias transvaginais. Os escores cervicais de cristalização e elasticidade foram associados aos níveis plasmáticos de estradiol. Mostrou-se que a concentração de estradiol necessária para aumento do escore da elasticidade é maior do que a necessária para produzir variação no escore da cristalização do muco cervical. Como a ocorrência do pico de LH, necessário à ovulação, exige níveis altos de estradiol, o escore da elasticidade tem maior associação com a ovulação do que o escore da cristalização.

Estudo prospectivo randomizado entre doxiciclina e oflaxacina no tratamento da uretrite näo-gonocócica causada pela chlamydia trachomatis / A prospective randomized trial of ofloxacin vs. doxycycline in the treatment of nongonococcal urethritis caused by chlamydia trachomatis

Neste estudo, 44 pacientes do sexo masculino com uretrite näo-gonocócica, causada por C. trachomatis, foram tratados randomizadamente segundo três protocolos clínicos. Quatorze pacientes receberam 200mg de ofloxacina de 12 em 12 horas por sete dias. Quatorze pacientes receberam 400mg de ofloxacina de 12 em 12 horas por sete dias. Dezesseis pacientes receberam uma dose inicial de 200mg de doxiciclina seguida de 100mg de 12 em 12 horas por sete dias. Após o tratamento, constatou-se que houve o desaparecimento da C. trachomatis em todos os pacientes que receberam ofloxacina porém, no grupo que recebeu doxiciclina, a cura bacteriológica näo ocorreu em dois pacientes (12,5 por cento). Quanto à tolerância ela foi considerada satisfatória nos três grupos