Rotina para o exame do semen: resultados encontrados em 30 exames normais / Basic routine for semen analysis: results found in 30 normal examinations

Uma rotina para o exame do semen (espermograma), adaptada a partir dos procedimentos tecnicos recomendados pela Organizaçao Mundial de Saude e descrita neste estudo. Esta rotina abrange o exame fisico (coagulaçao, tempo de liquefaçao, volume ejaculado, cor-aspecto, viscosidade e pH), dosagens bioquimicas (fosfatase acida e frutose), a analise das caracteristicas gerais dos espermatozoides (contagem celular, viabilidade, motilidade e morfologia) e a pesquisa de anticorpos antiespermatozoides. Usando esta rotina, resultados encontrados em 30 semens normais sao apresentados

Ausencia funcional de conductos eyaculadores / Functional absence of ejaculatory ducts

Se realizó un estudio retrospectivo de muestras de semen procedentes de 5 159 pacientes que se consultaron por infertilidad en un período de 8 años y 4 meses. En 188 casos se comprobó azoospermia (muestras seminales con ausencia de espermatozoides) y en 23 de los azoospérmicos se presentaron características seminales del síndrome de ausencia funcional de los conductos eyaculadores. Se compararon los valores bioquímicos del plasma seminal (fructosa, fosfatasa ácida, y glicerilfosforilcolina) de los pacientes con las características del síndrome con igual número de normozoospérmicos infértiles y azoospérmicos no obstructivos. Pudo comprobarse diferencia altamente significativa en los 3 indicadores (p < 0,005) entre los pacientes con el sídrome y los otros 2 grupos. Los resultados indican que estas determinaciones bioquímicas son útiles en el diagnóstico del síndrome sin necesidad de utilizar métodos quirúrgicos

Actividad de sialiltransferasa en la membrana espermática y su relación con fertilidad y esterilidad humanas / Sialyltransferase activity on spermatic membrane and its relationship with human fertility and sterility

La actividad de la sialiltransferasa (ST) fue determinada en los espermatozoides (spz) de sujetos normospérmicos (>80 X 10 6 spz/ml y mn[as del 75% de movilidad) y en los spz de pacientes con problemas de fecundidad (oligospérmicos < 20 y 10 6 spz/ml y menos del 20% de movilidad y astenospérmicos > 40 X 10 6 spz/ml y menos del 10% de movilidad). La actividad de la ST se cuantifica mediante la transferencia de radiactividad de CMP -3 H- siálico hacia el aceptor exógeno (fetuina desializada). El complejo enzima sustrato formado, al ser colocado en presencia del ácido fosfotúrgstico da un producto de fosfotungstato insoluble, el cual es retenido en un filtro de fibra de vidrio. La actividad enzimática disminuye en los spz de oligospérmicos en un 62 + - 5% con respecto a los spz de normospérmicos. La disminución de la actividad de la ST en los spz de pacientes infértiles permite suponer que esta enzima participa probablemente como causa directa de su patología y que su disminución obedece a un daño en la integridad estructural de la membrana espermática.(au)

Efecto inhibidor del plasma seminal humano sobre la proliferación linfocitaria in vitro inducida por fitohemaglutinina (PHA) / Inhibite effect of human seminal plasma on in vitro lymphocytic proliferation induced by phytohematogglutinin (PHA)

Se estudió el efecto de diferentes muestras de plasma seminal de hombres infértiles a diferentes concentraciones y de fracciones de diferente peso molecular de un pool de plasma seminal sobre la proliferación linfocitaria inducida por PHA. Los experimentos se realizaron en presencia de suero humano AB y los resultados mostraron un efecto el cual fue reproducible y dependiente de la concentración de plasma seminal. Varias fracciones mostraron efecto inmunosupresor, la de peso molecular entre 1 000 y 5 000 fue la de mayor potencia inhibidora

Correlaçao entre histologia, volume testicular e analise seminal em portadores de varicocele / Correlation among histology, testicular volume and semen analysis in patients with varicocele

Realizou-se a correçao cirurgica de varicocele e a biopsia testicular bilateral em 20 pacientes masculinos; avaliou-se a histologia espermatica, diametro e area tubular atraves da fotomicrografia. Dentre 20 pacientes, 11 apresentavam o testiculo esquerdo com volume pelo menos 25 por cento menor que o testiculo direito. Nos pacientes com menor volume testicular, nao houve diferença no diametro e area tubular, tendo porem menor numero de espermatides maduras por unidade tubular; esta diferença nao foi observada nos pacientes com igual volume testicular. Esses fatos nos permitem supor que a diminuiçao do volume testicular se faz as custas do espaço intersticial, provocando disfunçao das celulas de Leydig, que estao relacionadas com o numero de espermatides maduras, principal marcador da espermatogenese testicular

Concentration of ofloxacin and ciprofloxacin in human semen following a single oral dose.

A single oral dose of ofloxacin (400 mg.) or ciprofloxacin (500 mg.) was administered to each of 40 men. The mean concentrations of ofloxacin in semen exceeded those of ciprofloxacin at 1 hour (p = 0.035) and 24 hours (p = 0.028), while the levels of the 2 drugs at 2 and 4 hours were comparable. Semen concentrations of ofloxacin but not ciprofloxacin exceeded the reported minimal inhibitory concentration of Enterobacteriaceae, Chlamydia trachomatis and genital Mycoplasma species 24 hours after dosage.

The high rate of noncompliance for post-vasectomy semen examination: medical and legal considerations.

The records of 1,029 consecutive patients undergoing bilateral vasectomy were reviewed. Of the 1,029 patients 375 (36%) never returned postoperatively and the partners of 2 of these 375 later became pregnant. A total of 560 patients (54%) was followed until the recommended demonstration of azoospermia occurred on 2 consecutive occasions. There were 84 patients (8%) who did not return after the first demonstration of azoospermia, while 7 (1%) still had sperm in the semen at the last examination. Three other patients had persistent nonmotile sperm postoperatively; 2 of them underwent repeat vasectomy followed by azoospermia and 1 was placed in a special clearance category of sterility as described. We discuss practical and legal considerations regarding the management of noncompliant patients.

PIP: Urologists reviewed the medical charts of 1029 consecutive patients who had experienced a bilateral vasectomy (cauterization of the mucosa at each end and of the divided vas deferens) at the University of Louisville's School of Medicine in Kentucky between February and December 1986 to examine the medical and legal ramifications of vasectomized men not returning to have physicians examine their semen. Only 54.4% of the men returned for the recommended 2 semen examinations. 8.2% returned only once. 7 men (1% of the 654 who returned for semen examination) had sperm in their last semen sample. 36.4% (375) did not return at all after the vasectomy. Partners of 2 of these men conceived. 1 pregnancy occurred 29 months postoperatively. The men did not want to have his semen examined, however, for fear of learning he was sterile. The other pregnancy occurred 67 months after the vasectomy and the man's sperm had many motile sperm. Late recanalization of the vas deferens most likely occurred in this case. Other than recommending to patients before and during the vasectomy to submit 2 semen samples after the vasectomy, the urologists had no mechanisms in place to assure compliance. The high noncompliance rate in this study made the researchers consider their new legal responsibilities. Court cases for somewhat related medical situations have resulted in the jury finding the physician negligent because he/she did not fully inform the patient of the risk of noncompliance. Thus physicians, even though they may believe they are not responsible for noncompliance, should prudently inform patients of their risk of not complying by perhaps sending a punctually written notice. These warnings may protect them from potential litigation. Nevertheless the courts may appropriate more responsibility of patient actions to the physicians than is either morally justified or reasonable.

Seminal fluid from men with agenesis of the Wolffian ducts: zinc-binding properties and effects on sperm chromatin stability.

Zinc-binding properties were studied in 'prostatic fluid', i.e. in seminal plasma from patients with agenesis of the Wolffian ducts, and in split-ejaculate fractions dominated by seminal vesicular fluid. The effect of seminal fluid, with different zinc-binding properties, on the stability of zinc-dependent sperm chromatin was assessed by exposing sperm to 1% sodium dodecyl sulphate (SDS) for 60 min. Citrate was the only zinc ligand in 'prostatic fluid', as revealed by gel chromatography. Zinc in this fluid enhanced the stability of sperm chromatin. In contrast, the stability of sperm chromatin was decreased in seminal plasma dominated by vesicular fluid. These results are in accordance with the concept that prostatic fluid ensures the appropriate zinc content and stability of sperm chromatin, whereas abundance of vesicular fluid may jeopardize chromatin stability by reducing chromatin zinc content.

External quality control in the andrology laboratory: an experimental multicenter trial.

An external quality control study for semen analysis was performed involving 10 andrology laboratories in geographically separate locations. For the evaluation of sperm concentration, eight samples with different concentrations, and for the assessment of sperm morphology three slides prepared from different semen samples were distributed. Sperm motility was evaluated in five samples delivered cryopreserved to the participants. The coefficients of variation (CVs) for sperm counts varied with the sperm concentrations showing highest variability for low and lowest for high concentrations (range 23% to 73%). The CV for sperm morphology ranged from 25% for normal heads to 87% for abnormal midpieces. The CV for motility of sperm was 21%. For comparison the mean CVs for internal quality control were as follows: 10% for concentration, 8% for morphology (normal heads), and 8% for motility.

Sandwich ELISA assay for quantitative measurement of SP-40,40 in seminal plasma and serum.

SP-40,40 was purified from human plasma by PEG fractionation, DEAE-Sephacel, Phenyl-Toyopearl 650M, Bio-Gel A-0.5m and hydroxylapatite chromatographies. Three monoclonal antibodies (IF12, IID9 and IVF4) to this protein were prepared: IF12 and IID9 were specific for the beta subunit and IVF4 for the alpha subunit. The concentrations of SP-40,40 in seminal plasmas and sera were determined using a sandwich ELISA method. The results showed that the average concentrations of SP-40,40 were 438 +/- 285 micrograms/ml in seminal plasmas and 111 +/- 50 micrograms/ml in sera of normal donors. SP-40,40 concentrations in seminal plasmas of Klinefelter and excretory azoospermia patients were similar to those of normal donors. However, those of oligozoospermia and idiopathic azoospermia patients were about half the normal value.

Transrectal prostate ultrasonography as a diagnostic and therapeutic aid for ejaculatory duct obstruction.

Preoperative transrectal ultrasonography was used to establish the diagnosis of ejaculatory duct obstruction and also to determine the distal level of extension of the obstructed system within prostatic parenchyma. Transrectal ultrasonography not only established the diagnosis but also enabled precise transurethral resection into the obstructed system. Prostatic ultrasound facilitates evaluation and treatment of azoospermia caused by ejaculatory duct obstruction and may eliminate the need for vasography in such cases.

A sensitive enzyme-linked immunosorbent assay used to differentiate type AB specimen from a mixture of type A and type B specimens.

A sensitive enzyme-linked immunosorbent assay (ELISA) for detection of ABO blood substances in human body fluids was devised. The ELISA plates coated with purified human anti-A or anti-B serum effectively captured the blood substances, and these were then analysed by the combination of rabbit anti-A and goat anti-B. This capture ELISA could differentiate the type AB specimen from a mixture of the type A and the type B specimens, and the method was applied to rape cases to make the ABO typing of the criminal.

An investigation into the origin of the first fraction of the canine ejaculate.

Histological examination of the reproductive tract of the male dog failed to identify glandular tissue other than the prostate gland and its disseminated portion. A biochemical study of the three fractions of the ejaculate demonstrates similarities between the first and the third fractions. The composition of three ejaculates collected over 12 hours indicated that storage of prostatic secretion was a possible explanation for the biochemical differences observed. It is suggested that both the first and third fractions originate from the prostate gland.

Seminal transferrin and spermatogenic capability in the bull.

The objective of this study was to determine the relationship between seminal transferrin and sperm output in ejaculates from mature dairy bulls. Caudal sperm reserves in mature Holstein bulls (n = 15) were depleted by 8 successive ejaculations during a 50-70-min period (depletion phase). Bulls were then ejaculated 6 times per week for a period of 4 weeks (6X phase). Weekly sperm output (WSO) and weekly transferrin output (WTfO) were the sums of sperm and transferrin levels in 6 ejaculates taken in 1 week of the study. Mean WSO ranged from 20.7 billion to 39.6 billion and mean WTfO ranged from 334 micrograms to 1872 micrograms among the bulls. Regression analysis of sperm and transferrin levels in ejaculates collected during the depletion phase indicated that approximately 40% of seminal transferrin was not related to sperm output and probably was from accessory fluids. A relationship between total seminal transferrin and total sperm in ejaculate was observed (p less than 0.01, r = 0.54). This relationship was stronger when the transferrin was corrected for accessory fluid contribution (p less than 0.01, r = 0.65). The relationship between WSO and WTfO corrected for accessory fluid transferrin contribution (cWTfO) was significant (p less than 0.01, r = 0.64). The relationship between WSO and cWTfO can be interpreted to reflect the relationship between actual testicular sperm production and transferrin from testicular or epididymal origin.

Semen characteristics of asymptomatic males affected by Trichomonas vaginalis.

Trichomonas vaginalis is a sexually transmitted anaerobic parasite which causes vaginitis and cervicitis. The present study was carried out to determine the incidence of Trichomonas infection in semen samples of asymptomatic men and also to understand the changes of semen characteristics in the infected individuals before treatment and after treatment with metronidazole (Flagyl, 400 mg x 3x a day for 10 days). The semen obtained from 1131 men was examined for different characteristics and Trichomonas were detected in wet smear preparations in 50 cases (4.42%). The characteristics of semen in them was compared with 52 normal semen samples. Statistical analysis showed that the seminal fluid viscosity and percentage particulate debris was increased significantly in the infected group (P less than 0.001). There was no significant change in the pH of semen. Spermatozoan motility and morphologically normal forms were decreased significantly (P less than 0.001), spermatozoan viability was altered, and there was a significant change in membrane integrity (P less than 0.001) as indicated by the hypoosmotic swelling test. The significant improvement in semen characteristics were seen in 25 cases after a single course of treatment. Therefore, it is possible that in some cases, the infertility seen in asymptomatic individuals may be due to infection by Trichomonas.

Contraceptive potentiality of STS-557--a feasibility study in male bonnet monkey (Macaca radiata).

STS-557 (17 alpha-cyanomethyl-17 beta-hydroxy-estra-4, 9(10)-diene-3-one) was administered (i.m.) to two groups (4 in each group) of adult male bonnet monkeys at a daily dose of 10 mg/monkey for 12 weeks (first group) and 5 mg/monkey for 14 weeks (second group). Treatment with the 10 mg dose resulted in a significant decline in the count, motility, acrosin and hyaluronidase activities and the fertilizing ability (zona-free hamster egg penetration assay) of spermatozoa by the 6th week of initiation of treatment. The circulating level of STS-557 was low after one week and increased from the 2nd week of treatment when the serum level of testosterone was significantly reduced. Complete recovery was observed by the 11th week after withdrawal of treatment. The treatment with the 5 mg dose had minor and inconsistent effect on the motility, hyaluronidase and acrosin activity, and the fertilizing ability of spermatozoa in addition to the blood level of testosterone. STS-557 may have the potentiality to be used as a chemical contraceptive in the male but compensation for the reduced level of blood testosterone may be necessary.

Chlamydial infection--a female and/or male infertility factor?

After screening a large series (n = 491) of asymptomatic males of infertile partnerships for chlamydial immunoglobulin (Ig) G antibodies (Chlam AB), no significant influence of past chlamydial infection was found with regard to semen analysis, postcoital testing, in vitro sperm-cervical mucus penetration tests with hormonally standardized cervical mucus, circulating antisperm antibodies (detected with three different methods), local IgG and IgA antibodies (detected by means of the mixed antiglobulin reaction test) on the sperm surface, the sperm-cervical mucus contact test, and a microbial screening of semen samples for mycoplasmas and other potentially pathogenic micro-organisms. However, when the findings were correlated with infertility factors of patients' female partners and the subsequent pregnancy rate in a prospective study, a significant positive correlation of male Chlam AB with a tubal factor in their wives as cause of the couple's infertility was found. The results suggest that the main influence of Chlamydia trachomatis on male fertility is based on sexual transmission and negative influence on tubal function of female partners, but not on reduced sperm functional capacity.

The predictive value of zona-free hamster egg sperm penetration assay for failure of human in vitro fertilization and subsequent successful zona drilling.

The value of various sperm parameters and the zona-free hamster egg sperm penetration assay (SPA) in predicting human in vitro fertilization (IVF) failure and subsequent successful fertilization with zona drilling was assessed. In 19 couples, throughout 31 IVF cycles, a total of 153 oocytes failed to be fertilized. In subsequent 12 cycles with zona drilling, 33 of 131 (25%) were fertilized. The incidence of teratospermia and asthenospermia was significantly higher in the study group than in the control, 74% versus 32% and 42% versus 5%, respectively. Although the mean values for the performance of sperm in SPA and fertilization of human eggs after zona drilling were remarkably similar (28 +/- 6 versus 28 +/- 4), there was no correlation between individual parameters (r = 0.15). Thus, whereas male factor infertility is more likely to be associated with teratoasthenospermia, neither the SPA nor other sperm parameters have any predictive value for failure in IVF. In addition, no criterion of sperm function has yet been identified that would eliminate oligoteratoasthenozoospermic males from consideration of IVF with zona drilling.

Computerized slow-staged freezing of semen from men with testicular tumors or Hodgkin's disease preserves sperm better than standard vapor freezing.

Two methods of freezing semen taken from patients with testicular tumors or Hodgkin's disease before treatment were compared. Many patients already have semen abnormalities, so an optimal method is extremely important. Ejaculates from 8 patients with testicular tumors and 20 with Hodgkin's disease were frozen by fast-freezing or by slow-staged freezing. Effects of motility, viability, and swelling after thawing were significantly impaired with both methods. However, cryosurvival was better after slow- than fast-freezing: motility 24% +/- 12.4% versus 15% +/- 11.2%; viability 24.1% +/- 11.4% versus 17.3% +/- 10.4%, swelling 33.3% +/- 11% versus 27.6% +/- 12.8%. The effects were equal for normal and abnormal sperm. Sperm from tumor patients should be frozen by slow-staged freezing method in spite of the higher cost and longer time.

Eupergit C as a carrier for high-performance liquid chromatographic-based immunopurification of antigens and antibodies.

An immunoaffinity purification system using C30N and C1Z Eupergit C beads was developed and optimized. Poly- and monoclonal antibodies were purified using immobilized antigens and antigens were purified using immobilized antibodies. Antigens were used that possess enzymic activities and the efficiency of antigen binding was determined from the enzymic activity of the matrix-bound immunocomplexes. High-performance immunoaffinity purification using Eupergit C beads proved to be highly specific, reproducible, free from protein leakage and possessed a low degree of non-specific adsorption of tissue proteins. These characteristics of the system were illustrated by the isolation of immunoglobulin G from serum and of human decidua proteins from the decidua tissue and from seminal plasma. These proteins were obtained at high purity in a single purification step, as shown by sodium dodecyl sulphate-polyacrylamide gel electrophoresis.