Optimizing the transport of porcine semen: a proposal for Brazil / Otimizar o transporte do sêmen suíno: uma proposta para o Brasil

Semen from the first 15mL of the ejaculate (P1) obtained from two boars (30mL) was diluted in glycine-egg yolk extender, cooled at 5°C in a special container and rediluted in standard doses of 3x109 mobile spermatozoa after 12h of storage. Semen was also stored up to 24h after redilution. The physical characteristics of the semen were evaluated at different storage periods (fresh, 0h, 12h, rediluted, 24h, and 36h). The reproductive performance of the boars and their fertility regarding the insemination of primiparous sows were also determined. Two treatments were used: T1-15B sows inseminated with semen originated from hyperconcentrated heterospermic doses (15x109 mobile spermatozoa per dose), rediluted after 12h of storage at 5°C for standard doses of 3x109 mobile spermatozoa per dose and stored at 5°C up to 24h after redilution (n=10); T2-3B sows inseminated with standard heterospermic doses (3x109 mobile spermatozoa per dose), stored at 5°C up to 36h after semen collection (n=10). There was no effect (P>0.05) of treatments on the spermatic motility, even though a pronounced decrease (P>0.05) of their values at 12h of storage was recorded. However, they remained higher than 70% until 36h. There was effect of treatments on spermatic vigour at 0h (P<0.05), when T1-15B vigour was higher. There was also effect of the storage period for both treatments with a progressive decrease throughout 36h of storage, although the differences were not always significant. Pregnancy rates (90%) and the number of total farrowed piglets (15, 11-T1-15B; 13, 44- T2-3B) did not differ (P>0.05) between the treatments. It was concluded that the semen hyperconcentration of 15 billion of mobile spermatozoa per dose, stored at 5°C for 12h, did not result in drawbacks considering the physical characteristics of the semen, maintaining the pregnancy rates and prolificacy of the inseminated sows.

Os primeiros 15mL do ejaculado (P1) de dois varrões foram coletados (30mL) e diluídos em diluidor glicina-gema de ovo, resfriados a 5°C em contêiner especial e rediluídos para doses padrão de 3x109 espermatozoides (sptz) móveis, após 12 horas de armazenamento. Além disso, foram armazenados por até 24 horas após a rediluição, sendo as características físicas avaliadas em diferentes períodos de estocagem (fresco, zero hora, 12h, Red12h, 24h e 36h) e a fertilidade avaliada por meio de fêmeas primíparas inseminadas. Foram realizados dois tratamentos: T1-15B: porcas inseminadas com sêmen de doses heterospérmicas hiperconcentradas (15x109 sptz móveis/dose), rediluídas após 12 horas de armazenamento a 5°C para doses padrão de 3x109 sptz móveis/dose, e armazenadas a 5°C por até 24 horas após a rediluição (n=10); T2-3B: porcas inseminadas com doses heterospérmicas padrão (3x109 sptz móveis/dose), armazenadas a 5°C por até 36 horas após coleta. Não houve efeito (P>0.05) dos tratamentos sobre a motilidade espermática e, embora tenha ocorrido queda (P<0.05) às 12 horas, a motilidade foi superior a 70% durante as 36 horas de armazenamento. Houve efeito (P<0.05) dos tratamentos no tempo zero hora quanto ao vigor espermático, sendo E1T1-15B superior. Além disso, houve efeito do período de estocagem para os dois tratamentos, com queda progressiva do vigor ao longo das 36 horas, embora nem sempre as diferenças tenham sido significativas. As taxas de gestação (90%) e o número total de leitões nascidos (15, 11 - T1-15B; 13, 44 - T2-3B) não diferiram (P>0.05) entre os tratamentos. Concluiu-se que a hiperconcentração do sêmen para 15x109 sptz móveis/dose, armazenado a 5°C por 12 horas não resultou em prejuízos quanto à manutenção das características físicas do sêmen e ao desempenho reprodutivo dos varrões, sendo capaz de manter a taxa de gestação e a prolificidade das fêmeas inseminadas.

Insights into the role of cervical mucus and vaginal pH in unexplained infertility / Sobre o papel do muco cervical e do PH vaginal na gênese da infertilidade inexplicada

Unexplained infertility diagnosis is made in the presence of a normal semen analysis when tubal patency and normal ovulatory function are established. Among several potential causes, unexplained infertility could be attributed to vaginal pH and cervical mucus abnormalities. Although the vaginal canal and the cervix generally function as effective barriers to sperm, and although the production of mucus is essential to transport them from the vagina to the uterine cavity, these factors receive little attention in the investigation of couples with unexplained infertility. A substantial reduction in sperm number occurs as they transverse the cervix. From an average of 200 to 300 million sperm deposited in the vagina, only a few hundred achieve proximity to the oocyte. Given this expected high spermatozoa loss, a slight modification in cervical mucus may rapidly transform the cervix into a "hostile" environment, which, together with changes in vaginal environment and cervix structure, may prevent natural conception and be a cause of infertility. In this review, we discuss the physiological role of the vaginal pH and cervical mucus in fertility, and describe several conditions that can render the cervical mucus hostile to sperm and therefore be implicated in the pathophysiology of unexplained infertility.

RESUMO O diagnóstico de infertilidade inexplicada baseia-se na presença de espermograma normal, constatadas também permeabilidade tubária e função ovulatória normais. Entre as várias causas potenciais de infertilidade inexplicada, a presença de muco cervical e pH vaginal anormais devem ser consideradas. Embora a produção adequada de muco cervical seja essencial para o transporte dos espermatozóides da vagina para a cavidade uterina, e tanto o canal vaginal quanto o colo do útero desempenham função importante como barreira à passagem dos espermatozóides, estes fatores recebem pouca atenção na investigação de casais com infertilidade inexplicada. Uma redução substancial do número de espermatozoides ocorre à medida que estes percorrem o trato reprodutivo feminino. Partindo de cerca de 200 a 300 milhões de espermatozoides depositados na vagina, apenas algumas centenas alcançam a proximidade do oócito. Alteracões do muco cervical podem rapidamente transformar o colo do útero num ambiente hostil, que em conjunto com alterações no ambiente vaginal e da estrutura de colo do útero, podem apresentar-se condicões impedientes para a concepção natural; desse modo, convertem-se em causa de infertilidade. Nesta revisão, discutimos o papel fisiológico do pH vaginal e do muco cervical na fertilidade, descrever várias condicões que podem tornar o muco cervical hostil aos espermatozoides e, por fim analisamos como estes fatores interferem na fisiopatologia da infertilidade inexplicada.

Effects of feminine cleanser Inclear on sperm motility: A prospective study / 대한생식의학회지

OBJECTIVE: The objective of this study is to estimate the effects of Inclear, a feminine cleanser, on sperm motility. METHODS: Semen samples were obtained from infertile male patients. Following liquefaction, the raw semen samples were diluted with Ham's F-10 nutrient mixture medium containing 0.4% human serum albumin solution at a ratio of 1:3. The semen samples were subsequently centrifuged to separate the seminal plasma from the serum. The supernatant was discarded, and the pellet was resuspended. The sample was again centrifuged to remove cell debris, and the supernatant was removed. The final pellet was gently loosened by resuspension and incubated in medium alone as a control, and in a 10% solution of the medium plus Inclear. A sampling time of 30 minutes was selected on the basis of sperm transport studies. Sperm motility was evaluated with computer-assisted sperm analysis. RESULTS: A total of 20 samples were analyzed. The mean age of patients was 34.40+/-2.96 years. There was no difference in sperm concentration and motility in the two samples at 0 minute and 30 minutes of incubation. In both semen samples, the sperm concentration and motility decreased after an incubation period of 30 minutes. However, there was no statistical difference between the samples. Sperm concentration and motility were not significantly different between the control and Inclear samples after 0 minute and 30 minutes of incubation. CONCLUSION: Inclear has no negative effects on sperm motility. This product can be recommended to pregnancy planners for vaginal hygiene and as a vaginal lubricant.

Research progress in sperm mediated gene transfer technology / 生物医学工程学杂志

With the rapid development of biotechnology, we can change the trait of organism using transgenetic technology. In recent years, there are growing interests in the establishment of sperm mediated gene transfer (SMGT) technology as an effective and convenient method to produce transgenic animals. SMGT technology is a transgenetic method, which is easy in operation and does little harm to the cell compared with the other transgenetic methods. In this review, we expound the background, development, mechanism, operation and application of SMGT.

Morphological and morphometrical assessment of spermathecae of Aedes aegypti females

The vectorial capacity of Aedes aegypti is directly influenced by its high reproductive output. Nevertheless, females are restricted to a single mating event, sufficient to acquire enough sperm to fertilize a lifetime supply of eggs. How Ae. aegypti is able to maintain viable spermatozoa remains a mystery. Male spermatozoa are stored within either of two spermathecae that in Ae. aegypti consist of one large and two smaller organs each. In addition, each organ is divided into reservoir, duct and glandular portions. Many aspects of the morphology of the spermatheca in virgin and inseminated Ae. aegypti were investigated here using a combination of light, confocal, electron and scanning microscopes, as well as histochemistry. The abundance of mitochondria and microvilli in spermathecal gland cells is suggestive of a secretory role and results obtained from periodic acid Schiff assays of cell apexes and lumens indicate that gland cells produce and secrete neutral polysaccharides probably related to maintenance of spermatozoa. These new data contribute to our understanding of gamete maintenance in the spermathecae of Ae. aegypti and to an improved general understanding of mosquito reproductive biology.

Mecanismo de accion del levonorgestrel en la anticoncepcion de emergencia

En esta revisión se actualiza el mecanismo de acción del levonorgestrel (LNG) usado en anticoncepción de emergencia. El análisis crítico de la estimación de la eficacia anticonceptiva del LNG indica que su tasa de falla es más alta que la publicada. El LNG aumenta la viscosidad del moco cervical impidiendo que los espermatozoides del reservorio cervical vayan a renovar la población espermática en el sitio de fecundación. Diversos autores han documentado que LNG suprime el pico preovulatorio de gonadotrofinas e interfiere con el proceso ovulatorio en la mujer y en modelos animales. Administrado después de la fecundación en rata y Cebus apella, no interfiere con la implantación del embrión. Se concluye que LNG previene el embarazo solamente cuando se administra en un momento del ciclo menstrual en el cual puede impedir la fecundación y que el método falla cuando la administración es más tardía.

Would mirosurgical testicular sperm extraction replace the conventional technique for non-obstructive azoospermia?

To study the effectiveness of microsurgical testicular sperm extraction in non-obstructive azoospermia. Testicular sperm extraction [TESE] is well-defined procedure for surgical sperm retrieval in obstructive and non-obstructive azoospermia. This study was focused on the evaluation of the effectiveness of microsurgical testicular sperm extraction [TESE] in intracytoplasmic sperm injection [ICSI] for non-obstructive azoospermia. Material and Forty-two [42] male patients were included in this study. They were classified into 4 groups. Group I: Patients with normal sized tests and normal FSH, with previous diagnostic biopsy [10 patients]. Group II: Patients with normal sized tests and normal FSH, with previous conventional TESE [10 patients]. Group III: Patients with small sized tests and high FSH, and without previous testicular biopsy [20 patients]. Group IV: Patients with kleinfelter's syndrome [2 patients]. Microsurgical TESE was compared with conventional TESE as regard sperm recovery rate and complications were assessed by ultrasonographic and endocrinological methods. Microsurgical technique could extract sperm in 7 patients [70%] of group 1, 6 patients [60%] of group II and in in 5 patients [50%] of group III. It also succeded to extract sperm in group IV [kleinfelter's syndrome]. The sperm retrieval rate by microdissection TESE [60%] appeared higher than by conventional TESE [40%]. However, this observation failed to reach statistical significance. Microsurgical TESE could retrieve sperm in 100% of patients with hypospermatogenesis while conventional TESE retrieved sperm in 87.5%. Microsurgical TESE could retrieve sperm in 58.3% of patients with Sertoli only syndrome while conventional TESE retrieved sperm in only 8.3%. This was statistically significant [p < 0.05]. Microsurgical TESE could retrieve sperm in 20% of patients with maturation arrest while non-microsurgical TESE retrieved sperm in 40%. The complication rate was indifferent between microdissection and conventionol TESE. Microsurgical testicular sperm extraction is an ideal methodss for harvesting the sperm specially in difficult cases [small sized tests, cases of failed previous diagnostic biopsy or conventional TESE, repeated TESE, and in Kleinfelter's syndrome]. It allows selection of seminiferous tubules more likely to contain sperm specially in Sertoli only syndrome and Kleinfelter's syndrome. It also allows identification and avoidance of tessticular vessels, minimizing complications

addition of clonidine or fentanyl to lidocaine in local anaesthetic technique for testicular sperm extraction [TESE] a comparative study

A group of forty-five ASA I or II patients scheduled for testicular sperm extraction under local analgesia and sedation were studied. In order to improve the efficacy of local anaesthetic drug and better postoperative pain control, clonidine or fentanyl were added to lidocaine. The patients were randomly allocated into three equal groups. All patients received I.V midazolam and pethidine for sedation and glycopyrrolate premedication. In control group, lidocaine was used alone for local technique, however clonidine was added to lidocaine in the second group and fentanyl was added to lidocaine in the third group. Pulse and blood pressure were determined. Sedation and postoperative analgesia were assessed by subjective scoring. All groups showed haemodynamic stability. The mean sedation score was significantly higher in clonidine and fentanyl groups, the postoperative pain intensity and analgesic requirements were significantly less in clonidine group than in control and fentanyl groups. These results confirm that adding clonidine to lidocaine provides an effective safe analgesia for TESE with better postoperative analgesia and early ambulation

Spermatotoxic effect of carbendazim.

Carbendazim, suspended in sunflower oil, was administered to Wistar male rats through an oral intubation at a daily dose of 25 mg/kg body weight for 48 days, and the cauda epididymal sperm were analysed on day 49 for counts, motility and abnormalities. The study indicates that carbendazim affects the cauda epididymal sperm as seen in decreased sperm counts, inhibition of motility and increased incidence of abnormalities.

Uso de midodrín y congelación de semen en el tratamiento de las alteraciones del transporte espermático: caso clínico / Use of midodrine and frozen/thawed semen to treat semen transport disturbances: report of 2 cases

Retrograde ejaculation severely compromises male fertility. The use of sympathicomimetics for the treatment of this condition has poor results, except in patients with partial retrograde ejaculation, whose semen has a higher spermatozoa concentration. The semen of two patients with partial retrograde ejaculation was collected and frozen after the injection of a sympathicomimetic (Midodrine). The frozen/thawed samples were mixed with fresh semen recently ejaculated to obtain a minimal number of motile spermatozoa, and used for intrauterine insemination (> de 1 x 106 motile spermatozoa/ml). In both cases, pregnancies that developed satisfactorily, were obtained

La región epididimaria de la paloma (columbia livia): análisis morfológico y morfométrico / The epididymal region of the pigeon (columba livia): morphological and morphometrical evaluation

La vía espermática extratesticular de la paloma está formada por los conductores eferentes proximales y distales, y Por el conducto epididimario. Los conductos eferentes están revestidos por epitelio pseudo-estratificado, estereocilado y el conducto Epididimario por epitelio columnar bajo, pseudo-estratificado. El análisis morfométrico demostró que los conductores eferentes proximales Poseen mayor diámetro medio entre los túbulos de la región epididimaria (X: 283,64 um); teniendo este hecho una probable implicancia Fisiológica en el proceso de reabsorción del fluido seminífero

Changes of Contractility of the Vas Deferens to Field Stimulation in Diabetic Rats / 대한비뇨기과학회지

Diabetes is known to induce autonomic dysfunction and most experiments have been focused on the smooth muscle dysfunctions of the urinary bladder and corpus cavernosum in the genitourinary tract. The contractile response of the vas deferens is also decreased in diabetic rats and this effect may cause impairment of sperm transportation. However, few studies have been investigated the changes of contractility of the vas deferens in diabetic animals and little attention has been given to the differences between the contractilities of the prostatic and epididymal ends of the vas deferens in diabetic rats. In this study, the whole vas deferens was divided into two portions, prostatic and epididymal ends, and we examined the contractility of both segments to field stimulation at various frequencies from 4 to 64 Hz for 30 sec with pulses of 1 msec duration at 80 V in organ bath containing modified Tyrode's solution. Both initial twitch and the last contraction after 30 sec elicited by field stimulation were compared to clarify which component was more susceptible to field stimulation in control and diabetic rats. In the prostatic end, initial twitch was more impaired than the last contraction, which is believed that in diabetic rats, ATP(adenosine triphosphate) is more affected than NA(noradrenaline). In contrast, in the epididymal and the last contraction was more impaired than the initial twitch. In 64 Hz, initial twitch and the secondary contractions were not distinguished in the epididymal end of control rats, but in diabetic rats both phases were distinct. This means that the release of NA to field stimulation in the epididymal end is delayed in diabetic rats.

Transporte de gametas / The gametes migration

Apresenta-se uma revisäo sobre transporte de gametas.

Human pregnancy following sperm antibody separation technique and sperm intra-uterine transfer

Microscopic examination of the seminal fluid of infertile patients who gave positive kibrick and/or F-D tests also revealed massive sperm clumping and agglutination. Positive correlation was observed between infertility and sperm antibodies. Sperm antibodies were also found in the cervical mucus of infertile females. Sperm clumping was treated by dilution with 50% fetal calf serum followed immediately. by physical and mechanical separation of sperm clumping several times. The sperm motility was increased from 10% to 50%. Ovarian stimulation was induced by clomipgene citrate [100 mg from day two to day six of the cycle] and human menopausal gonadotropin [two ampoules from day 7 to 11 of the cycle] followed by 5000 to 10000 International Units [IU] of Human chorionic gonadotropin when the diameter of the ovarian follicles was more than 1.8cm and the concentration of estradiole was more than 300pg/ml. Following the separation of sperm clumping, the hyperactive sperm cells were selected through a simple layer technique. A concentration of one to two millions of sperm cells was removed from the top and middle rich sperm fraction and transferred in less than 200 micro liter of sperm culture medium into the uterus by special catheter. A total of ten transfers was done and five pregnancies induced. Out of five pregnancies, three were aborted within the first trimester and two pregnancies gave birth. The first baby was a male born on 31 of January and the second case was a female born on first of February 1990. Both babies are healthy and weighing more than 3000 grams and the lengths are also more than 49cm. It was concluded from the results of this study that the technique used under the experimental conditions of the present study was found to have a significant value in the treatment of infertile couples

Phosphoinositide specific phospholipase C activity of goat spermatozoa in transit from the caput to the cauda epididymis.

Goat sperm lysate from cauda epididymis was incubated in the presence of [14C]phosphatidyl-choline, -ethanolamine, -inositol and diphosphatidyl-glycerol. The release of [14C]diacylglycerol from only phosphatidyl inositol confirmed the presence of phosphatidyl inositol specific phospholipase C. The enzyme activity was linear up to 1 hr of incubation at a sperm concentration of 1-10 x 10(9). It had pH optimum of 7.5 in a broad range of pH activity profile (pH 6-9). Maximum activity was observed at 8 mM calcium ion concentration. EDTA and EGTA (5 mM) did not inhibit the activity completely. A comparative study on spermatozoa and fluid from caput, corpus and cauda epididymis revealed 6.5-fold increase of activity in spermatozoa and a 4-fold decrease in case of fluid during the epididymal transit. However, the total protein content remained unchanged in fluid and decreased up to the extent of 2.4-fold in spermatozoa during this process. Thirty five percent of the caudal sperm activity was soluble and the rest was associated with head (44%), mid-piece (10%) and tail (10%).

Teste de Huhner ou teste de Sims-Huhner? / Huhner or Sims-Huhner test?

Descreve-se de modo resumido a biografia de J. M. Sims. Mostra-se o processo que o ilustre médico trouxe à cirurgia e ginecologia dos Estados Unidos da América, razäo por que é denominado o "Pai da Ginecologia Norte-americana". Além da magistral contribuiçäo ao tratamento das fístulas vesicovaginais, preocupou-se com o problema da infertilidade e descreveu com detalhes o teste pós-coito em 1866. Apesar de merecidos elogios pelo desempenho profissional, foi considerado pessoa insensível por realizar repetidas operaçöes em escravas negras e em imigrantes pobres de Nova Iorque, no intuito de aperfeiçoamento cirúrgico. Mas Hühner, em 1913, publicou um livro em que expôs os diferentes exames a serem realizados na pesquisa diagnóstica do casal infértil e redescreveu o teste pós-coito. Em 1929 o livro teve uma segunda ediçäo. Em artigo de divulgaçäo de 1924, Hühner enumerou os exames descritos anteriormente e mencionou que médicos daquela época tinha proposto a denominaçäo de "teste de Hühner" ao exame pós-coito por ele relatado como "teste do espermatozóide". Na bibliografia do artigo há citaçäo de um trabalho, posterior ao de 1866, é bem explícito e, além da descriçäo da prova, realça o valor do microscópio e assinala as condiçöes adversas ao teste. Causa estranheza a omissäo de Hühner

Prueba de integridad funcional de la membrana espermatica: valor en el estudio del hombre infertil / Functional integrity test for the sperm membrane: the value in the study of interfile men

Se analiza el valor diagnóstico de la PIFME en 51 pacientes infértiles según la técnica descrita por Jeyedran (1984) clasificándolos en tres grupos: A) normozoospérmicos, B) oligozooespérmicos, C) astenozoospérmicos. Al comparar los valores medios del % de PIFME en cada grupo se encontraron diferencias significativas entre A y B entre B y C, pero no así entre A y C. De acuerdo a la movilidad, Ct/ml y morfología basal, los pacientes se agruparon atendiendo al criterio de normalidad de la OMS, hallándose diferencia estadísticamente significativa (p<0,02) entre los subgrupos de cada indicar. El índice de correlación encontrado entre el % de PIFME vs movilidad, Ct/ml y morfología en la muestra total fue de 0,33, 0,52 y 0,50 respectivamente. En el grupo de los normozooespérmicos no se encontró correlación significativa alguna, mientras que en los oligozoospérmicos se halló solamente con la movilidad (r = 0,61) ambos casos positiva y significativa. Se concluye que la PIFME debe ser considerada como un indicador más del análisis seminal sistemático, ya que su realización no es compleja, no eleva el costo de la investigación y permite precisar otra posible causa de disminución de la capacidad fecundante del espermatozoide

Epididymal transit and morphology of the hamster sperm head

Los cambios que experimenta la cabeza del espermatozoide de hamster durante su tránsito por el epidídimo fueron analizados con técnicas morfológicas, de criofractura y morfométricas. Desde el punto de vista morfológico se comprueba una modificación del ángulo formado por el acrosoma y la región postacrosomal que de recto en espermatozoides del caput pasa a obtuso en espermatozoides de cauda. En cuanto a la criofractura se estudió la distribución de las partículas intramembranosas en la membrana plamática (IMPS). Se describe una disposición regular de las IMPS formando una trama hexagonal en el borde convexo del acrosoma. Esta trama está presente en espermatozoides extraídos del caput y no se observa en los extraídos de la región caudal del epidídimo. La región postacrosomal revela dos cambios notables: la presencia de una corona de barras adyacentes al cuello y formada por IMPS de 10 nm en espermatozoides del caput que se desorganiza en espermatozoides extraídos del epidídimo caudal y el incremento en estos del número de IMPS de 10 nm dispersas por toda la región postacrosomal. Postulamos que el aumento de IMPS en la región postacrosomal se relaciona con la incorporación de glicoproteínas secretadas por epidídimo como factor (es) de maduración y reconocimiento intergamético