Presence Of Sperm Research Articles

A Transport Model for Estimating the Time Course of ERK Activation in the C. elegans Germline

The Caenorhabditis elegans germline is a well-studied model system for investigating the control of cell fate by signaling pathways. Cell signals at the distal tip of the germline promote cell proliferation; just before the loop, signals couple cell maturation to organism-level nutrient status; at the proximal end of the germline, signals coordinate oocyte maturation and fertilization in the presence of sperm. The latter two events require dual phosphorylation and activation of ERK, the effector molecule of the Ras/MAPK cascade. In C. elegans, ERK is known as MPK-1. At this point, none of today’s methods for real-time monitoring of dually phosphorylated MPK-1 are working in the germline. Consequently, quantitative understanding of the MPK-1-dependent processes during germline development is limited. Here, we make a step toward advancing this understanding using a model-based framework that reconstructs the time course of MPK-1 activation from a snapshot of a fixed germline. Our approach builds on a number of recent studies for estimating temporal dynamics from fixed organisms, but takes advantage of the anatomy of the germline to simplify the analysis. Our model predicts that the MPK-1 signal turns on ∼30 h into germ cell progression and peaks ∼7 h later.

Morphological analysis of the oviduct, oviducal gland and isthmus of the blue shark Prionace glauca (Linnaeus, 1758) (Elasmobranchii: Carcharhiniformes)

ABSTRACT Oviducal gland present in elasmobranchs is correlated to the organism's reproductive strategy, and its functions are to produce mucus, to form the egg's tertiary envelope and to store sperm. The gland contains four zones: club, papillary,baffle and terminal. The structures of the oviduct, oviducal gland and isthmus of blue shark Prionace glauca were described using macroscopic, light microscopy and scanning electron microscopy techniques. The epithelium of the oviduct and isthmus is folded and is a simple, columnar, ciliated lining epithelium with glandular cells. In the oviducal gland, the lining tissues in the four zones are similar to the oviduct and isthmus lining. The terminal zone shows the presence of sperm in the lumen of the secretory tubules, which remains stored even in the absence of recent copulation. Here, these organs were studied and their connections in an attempt to elucidate the mechanisms of reproduction in the blue shark, showing the three-dimensional aspects, thus adding morphological information important for the understanding of the structure and functioning of these organs of fundamental importance in the life of the majority of elasmobranchs.

Genetically determined patozoospermia. Literature review and research results

Genetic factors (chromosomal aberrations and point mutations) are the cause of infertility in 10–15 % of men with impaired fertility. Homogeneous structural and functional defects in the sperm or the total terato-, asthenozoospermia – rare cases of genetically determined male infertility, are autosomal recessive diseases. Currently, described 4 types of «syndromic» spermopatology. 1. Primary ciliary dyskinesia (PCD) in men with total asthenozoospermia. Affects axoneme structures (microtubules, dynein arms, radial spokes). It identified more than 20 chromosomal loci responsible for the development of the PCD. 2. Dysplasia of the fibrous sheath of sperm tail in men with asthenozoospermia. The shortened and thickened sperm tail observed with disorganization of vertical columns and cross ribs of the fibrous sheath. Candidate genes – genes family ACAP. 3. Globozoospermia in men with teratozoospermia characterized by the presence of sperm with round heads, primary lack of acrosome and disorganization middle part of the flagellum. Found mutations or deletions of genes SPATA16, PICK1 and DPY19L2. 4. Syndrome decapitated spermatozoa in men with teratozoospermia (microcephaly). Abnormalities in the spermiogenesis development of connecting part jf the tail and proximal (morphologically normal) centrioles. In 2012–2014 years we have studied the ultrastructure of 2267 semen samples of men with impaired fertility. Globozoospermia revealed in 7 patients, dysplasia of the fibrous sheath – 13, decapitated sperm – in one. PCD was revealed in 4 patients (lack of axoneme dynein arms was found in 1 patient, absence of axoneme radial spokes – in 3 patients. The problem of genetically determined patozoospermya must be taken into account when the assisted reproductive technologies practises. There are few cases of successful assisted reproductive technologies with sperm of these patients. We don»t know the etiological factors of syndromic spermopatologe, so we cannot determine the degree of genetic risk.

The Sertoli cell: Novel clinical potentiality.

The Sertoli cell is important for endocrine and paracrine control of spermatogenesis. Functions attributed to Sertoli cells are: (1) supportive and trophic functions for the cells of the seminiferous epithelium, (2) transport of mature spermatids towards the lumen of seminiferous tubules, (3) secretion of androgen binding protein, (4) production of substances with endocrine or paracrine action for spermatogenesis control and (5) interaction with intertubular endocrine Leydig cells. Inhibin B and anti-Müllerian hormone (AMH) are glycoproteins belonging to the transforming growth factor β (TGF-β) superfamily; they are produced almost exclusively by the Sertoli cells and have been proposed as direct markers of their function and indirect markers of spermatogenesis. Serum inhibin B and AMH concentrations seem to constitute additional diagnostic parameters in male subfertility as they reflect Sertoli cell function. Stimulated concentrations of serum inhibin B and AMH do not add clinically relevant information in subfertile men compared to basal concentrations of these hormones. Serum inhibin B and AMH concentrations correlate with testicular histology/cytology but are not superior to FSH as predictors of the presence of sperm in testicular sperm extraction (TESE)/fine needle aspiration (FNA) biopsy in men with azoospermia.

Sperm storage and morphofunctional bases of the female reproductive tract of the snake Philodryas patagoniensis from southeastern Brazil

Macroscopic and microscopic examination of the female reproductive tract of Philodryas patagoniensis was conducted in order to observe sperm storage structures and determine morphological changes throughout the reproductive cycle. The oviduct of P. patagoniensis is composed of the infundibulum (anterior and posterior), uterus, utero-vaginal junction, and vagina. The uterine epithelium showed constant secretory activity throughout the reproductive cycle, although increased production of secretory granules was observed in secondary vitellogenic females. Sperm storage occurs in the posterior infundibulum and utero-vaginal junction in pregnant, primary (postpartum), and secondary vitellogenic females. These data suggest that P. patagoniensis may be able to produce several clutches from a single mating. Sperm storage in the posterior infundibulum occurs in sperm receptacles, while in the utero-vaginal junction sperm is stored in crypts. Histochemical tests by periodic acid–Schiff revealed that both sperm storage sites increase the production of neutral carbohydrate-rich granules in the presence of sperm, similar to snakes from temperate regions. This is the first report of sperm storage in the posterior infundibulum of a neotropical snake.

Evolutionary Morphology of the Primary Male Reproductive System and Spermatozoa of Goblin Spiders (Oonopidae; Araneae)

Goblin spiders (Oonopidae Simon, 1890) are distributed worldwide and among the most species-rich spider taxa. However, goblin spiders are understudied in many aspects and their phylogenetic relationships are not well resolved. As previously shown for numerous other spider groups the male and female reproductive system bears many characters of phylogenetic relevance. Moreover, the diversity of sperm structures within spiders is astonishingly diverse and often taxon specific. In the present study, we analyzed the primary male reproductive system and spermatozoa of goblin spiders for the first time. We investigated 18 species of 13 genera representing the subfamilies Orchestininae and Oonopinae by means of light and transmission electron microscopy. We scored 44 characters from the gross morphology of the reproductive system as well as spermatozoa including four new characters for the male spider reproductive system. All investigated species transfer sperm as synspermia, a method corroborating with the recently proposed “Synspermiata” clade unifying all ecribellate Haplogynae. Furthermore, goblin spiders show by far the highest diversity of sperm structures in spiders. In total, we recovered 30 unambiguous synapomorphies for different oonopid taxa. In a comparison with all other spider taxa studied to date, we identified the longest sperm (Neoxyphinus termitophilus) and longest sperm conjugates (Orchestina). Moreover and most remarkable is the presence of aflagellate sperm in Opopaea apicalis, which is the first report of the loss of a sperm flagellum in tetrapulmonate arachnids. These findings are of high interest not only because of their phylogenetic implications, but also with regard to their contribution to our understanding of postcopulatory sexual selection in spiders.

Evidence for mate-encounter Allee effect in an invasive longhorn beetle (Coleoptera: Cerambycidae)

1. Limited empirical support is available for mate-encounter Allee effects in invasive insects due to the logistical challenges of studying demographic trends in low-density populations. 2. Traps baited with pheromone and spruce volatiles were used to monitor the abundance of female Tetropium fuscum F. (Coleoptera: Cerambycidae) at multiple sites in Nova Scotia in 2011 and 2012. Each female was dissected to determine the presence or absence of sperm in its spermatheca (mated or virgin female, respectively). 3. Both male and total T. fuscum abundance declined with increasing distance to the focal point of T. fuscum's invasion. Female mating probability declined with male abundance and with distance from the invasion focus, and mating probabilities were very low at the most peripheral sites. Difficulty in encountering mates may thus contribute to limiting the spread of T. fuscum. 4. The approach outlined here could be integrated into existing surveys of wood borers using traps baited with semiochemicals to improve our understanding of the role of the mate-encounter Allee effect in invasion dynamics.

Clinical Background of Patients with Sperm in Their Urinary Sediment.

IntroductionThe detection rate and associated factors of at least one sperm in urinary sediment is not well-known in real clinical practice.AimsThe aim of the present study was to evaluate the clinical features associated with the presence of sperm in urinary sediment in a large number of samples.MethodsWe conducted a cross-sectional study at Tokyo Saiseikai Central Hospital. We identified 5,005 males who were aged ≥20 years in whom urinary sedimentation had been performed at least twice between May 2011 and June 2012. The sperm group included patients in whom at least one urinary sediment test performed under a microscope had detected at least one sperm. We evaluated the associations between the presence of at least one sperm in urinary sediment and clinical parameters such as various diseases and the use of particular oral medicines.Main OutcomesIn total, 1.6% (339/20,937) of urinary sediment samples contained at least one sperm. The sperm group consisted of 282 subjects (5.6%), and the no-sperm group included 4,723 subjects (94.3%).ResultsMultivariate analysis demonstrated that younger age (<65) (odds ratio [OR]: 1.71, 95% confidence interval [CI]: 1.32–2.21), the total number of examinations (≥4) (OR: 1.46, 95%CI: 1.11–1.92), diabetes (OR: 1.72, 95%CI: 1.31–2.25), a history of pelvic surgery for colon cancer (OR: 4.89, 95%CI: 2.38–10.02), alpha-1 blocker use (OR: 1.55, 95%CI: 1.16–2.08), a history of trans-urethral resection of the prostate (OR: 2.77, 95%CI: 1.46–5.13), and selective serotonin reuptake inhibitor use (OR: 2.12, 95%CI: 1.07–4.19) were independent predictors of the presence of at least one sperm in urinary sediment.ConclusionThere is considerable overlap between the factors associated with the presence of at least one sperm in urinary sediment and those that are strongly associated with ejaculatory disorders.

Human sperm degradation of zona pellucida proteins contributes to fertilization

BackgroundThe mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. Although penetration of the ZP is a limiting event to achieve fertilization, this is one of the least comprehended stages of gamete interaction. Even though previous studies suggest that proteases of sperm origin contribute to facilitate the passage of sperm through the ZP, in human this process is not yet fully understood. The aim of this study was to determine the ability of human sperm to degrade recombinant human ZP (rhZPs) proteins and to characterize the proteases involved in this process.MethodsPurified rhZP2, rhZP3 and rhZP4 proteins were incubated with capacitated sperm and the proteolytic activity was determined by Western blot analysis. To further characterize the proteases involved, parallel incubations were performed in the presence of the protease inhibitors o-phenanthroline, benzamidine and MG-132 meant to block the activity of metalloproteases, serine proteases and the proteasome, respectively. Additionally, protease inhibitors effect on sperm-ZP binding was evaluated by hemizona assay.ResultsThe results showed that rhZPs were hydrolyzed in the presence of capacitated sperm. O-phenanthroline inhibited the degradation of rhZP3, MG-132 inhibited the degradation of rhZP4 and benzamidine inhibited the degradation of the three proteins under investigation. Moreover, hemizona assays demonstrated that sperm proteasome inhibition impairs sperm interaction with human native ZP.ConclusionsThis study suggests that sperm proteasomes could participate in the degradation of ZP, particularly of the ZP4 protein. Besides, metalloproteases may be involved in specific degradation of ZP3 while serine proteases may contribute to unspecific degradation of the ZP. These findings suggest that localized degradation of ZP proteins by sperm is probably involved in ZP penetration and may be of help in understanding the mechanisms of fertilization in humans.

Pubertal age based on testicular and epididymal histology in Göttingen minipigs

Göttingen minipigs are used worldwide as nonrodent animal models for toxicologic and pharmaceutical research. Having knowledge of the age for onset of puberty is an important consideration when designing such biomedical experiments. The present study reports an earlier age for puberty on the basis of changes in testicular spermatogenic development in the Göttingen minipig. Testes and epididymides of 24 Göttingen minipigs ranging 5 to 8 weeks of age were obtained for histologic observation after hematoxylin–eosin staining. Microscopic examination was performed to determine the prevalence of cell types (e.g., spermatogonia, primary and secondary spermatocytes, round spermatids, elongated spermatids, luminal sperm) in seminiferous tubules and for the presence of sperm in the cauda epididymis. Puberty was defined as having a majority cell type presence of elongated spermatids in the seminiferous tubules in conjunction with the presence of spermatozoa in the cauda epididymis. Puberty was identified in two males at 6 weeks of age, two males at 7 weeks of age, and in all 6 males at 8 weeks of age. In conclusion, the age of puberty in male Göttingen minipigs occurs by 8 weeks of age, an earlier age than previously reported.

Male infertility and its genetic causes.

Infertility is a serious social problem in advanced nations, with male factor infertility accounting for approximately half of all cases of infertility. Here, we aim to discuss our laboratory results in the context of recent literature on critical genes residing on the Y chromosome or autosomes that play important roles in human spermatogenesis. The PubMed database was systematically searched using the following keywords: 'genetics of male factor infertility'; 'male infertility genes', 'genetics of spermatogenesis' to retrieve information for this review. Striking progress has recently been made in the elucidation of mechanisms of spermatogenesis using knockout mouse models. This information has, in many cases, not been directly translatable to humans. Nevertheless, mutations in several critical genes have been shown to cause male infertility. We discuss here the contribution to male factor infertility of a number of genes identified in the azoospermia factor (AZF) region on the Y chromosome, as well as the autosomally located genes: SYKP3, KLHL10, AURKC and SPATA16. Non-obstructive azoospermia is the most severe form of azoospermia. However, the presence of spermatozoa can only be confirmed through procedures, which may prove to be unnecessary. Elucidation of the genes underlying male factor infertility, and thereby a better understanding of the mechanisms that cause it, will result in more tailored, evidence-based decisions in treatment of patients.

Oviductal Transcriptome Is Modified after Insemination during Spontaneous Ovulation in the Sow

Gene Expression Microarray technology was used to compare oviduct transcriptome between inseminated and non-inseminated pigs during spontaneous oestrus. We used an in vivo model approaching the study from a physiological point of view in which no hormonal treatment (animals were in natural oestrus) and no artificial sperm selection (selection was performed within the female genital) were imposed. It is therefore emphasised that no surgical introduction of spermatozoa and no insemination at a site other than the physiological one were used. This approach revealed 17 genes that were two-fold or more up-regulated in oviducts exposed to spermatozoa and/or developing embryos and 9 genes that were two-fold or more down-regulated. Functional analysis of the genes revealed that the top canonical pathways affected by insemination were related to the inflammatory response and immune system (Network 1) to molecular transport, protein trafficking and developmental disorder (Network 2) and to cell-to-cell signalling and interaction (Network 3). Some of the genes in network 1 had been previously detected in the oviduct of human and animals, where they were over-expressed in the presence of spermatozoa or pre-implantation embryos (C3, IGHG1, ITIH4, TNF and SERPINE1) whereas others were not previously reported (SAA2, ALOX12, CD1D and SPP1). Genes in Network 2 included RAB1B and TOR3A, the latter being described for the first time in the oviduct and clearly expressed in the epithelial cells of the mucosa layer. Network 3 integrated the genes with the highest down-regulation level (CYP51, PTH1R and TMOD3). Data in the present study indicate a change in gene expression during gamete encounter at the site of fertilization after a natural sperm selection within the female genital tract. These changes would indicate a modification of the environment preparing the oviduct for a successful fertilization and for an adequate embryo early development.

Procaine Induces Cytokinesis in Horse Oocytes via a pH-Dependent Mechanism.

Coincubating equine gametes in the presence of procaine has been reported to facilitate in vitro fertilization, with cleavage rates exceeding 60%. We report that while procaine does trigger sperm hyperactivation, it independently induces cleavage of equine oocytes. First, we found that procaine (1-5 mM) did not facilitate stallion sperm penetration of equine oocytes but instead induced sperm-independent oocyte cytokinesis in the absence of the second polar body extrusion. Indeed, 56 ± 4% of oocytes cleaved within 2.5 days of exposure to 2.5 mM procaine regardless of sperm presence. However, the cleaved oocytes did not develop beyond 8 to 16 cells, and the daughter cells either lacked nuclei or contained aberrant, condensed DNA fragments. By contrast, intracytoplasmic sperm injection (ICSI) was followed by second polar body extrusion and formation of normal blastocysts. Moreover, neither the calcium oscillations detectable using fura-2 AM staining nor the cortical granule reaction visualized by LCA-FITC staining, after oocyte activation induced by ICSI or ionomycin treatment, were detected after exposing oocytes to 2.5 mM procaine. Instead, procaine initiated an ooplasmic alkalinization, detectable by BCECF-AM staining that was not observed after other treatments. This alkalinization was followed, after an additional 18 h of incubation, by cortical F-actin depolymerization, as demonstrated by reduced actin phalloidin-FITC staining intensity, that resembled preparation for cytokinesis in ICSI-fertilized zygotes. Overall, we conclude that procaine induces cytokinesis in equine oocytes accompanied by aberrant chromatin condensation and division; this explains why embryos produced after exposing equine oocytes to procaine fail to develop beyond the 8- to 16-cell stage.

Postcopulatory sexual selection favors fertilization success of restocking hybrid quails over native Common quails (Coturnix coturnix)

Postcopulatory sexual selection plays an important role in the reproductive success of males in many species. Differences in fertilization success could affect rates of admixture and genetic introgression between divergent lineages. We investigated sperm precedence in matings in captivity involving Common quails (Coturnix coturnix) and farm quails of hybrid origin (C. coturnix × domestic Japanese quail, C. japonica), the last used in restocking practices to increase hunting bags. These inter-specific matings in natural conditions are claimed to represent an important threat to the conservation of native Common quail populations. Results showed that fertilization success of each male depended on (1) the time it spent with the female, (2) the presence of sperm from a previous male in the female oviduct, (3) the time that the previous partner had been copulating with the female, and, most importantly, (4) the genetic origin of the male (wild or farm). Farm hybrid males showed higher fertilization success than wild Common males, and they required less time with the female to fertilize the same proportion of eggs. The presence of sperm from another male in the female oviduct reduced the percentage of fertilized eggs by a male. However, this reduction was higher for wild males when the precedent mate was a farm male. In summary, the sperm of farm hybrid males may outcompete the sperm of native males and this could be favoring the introgression of domestic Japanese alleles into the Common quail population, thus constituting a severe conservation threat to wild Common quail populations.

Genetic aspects of monomorphic teratozoospermia: a review.

Teratozoospermia is characterized by the presence of spermatozoa with abnormal morphology over 85% in sperm. When all the spermatozoa display a unique abnormality, teratozoospermia is said to be monomorphic. Two forms of monomorphic teratozoospermia, representing less than 1% of male infertility, are recognized: macrozoospermia (also called macrocephalic sperm head syndrome) and globozoospermia (also called round-headed sperm syndrome). Macrozoospermia is defined as the presence of a very high percentage of spermatozoa with enlarged head and multiple flagella. Meiotic segregation studies in 30 males revealed that over 90% of spermatozoa were aneuploid, mainly diploid. Sperm DNA fragmentation studies performed in a few patients showed an increase in DNA fragmentation index compared to fertile men. Four mutations in the AURKC gene, a key player in meiosis and more particularly in spermatogenesis, have been found to be responsible for macrozoospermia. Globozoospermia is characterized by round-headed spermatozoa with an absent acrosome, an aberrant nuclear membrane and midpiece defects. The rate of aneuploidy of various chromosomes in spermatozoa from 26 globozoospermic men was slightly increased compared to fertile men. However, this increase was of the same order as that commonly found in infertile men with altered sperm parameters. The majority of the studies found that globozoospermic males had a sperm DNA fragmentation index higher than in fertile men. Mutations or deletions in three genes, SPATA16, PICK1 and DPY19L2, have been shown to be responsible for globozoospermia. Identification of the genetic causes of macrozoospermia and globozoospermia should help refine diagnosis and treatment of these patients, avoiding long and painful treatments. Elucidating the molecular causes of these defects is of utmost importance as intracytoplasmic sperm injection (ICSI) is very disappointing in these two pathologies.

Detection of insemination status in live Aedes aegypti females

Using the technique described in this report, the presence or absence of sperm in spermathecae of female Aedes aegypti is detectable without dissection. Spermathecae of a lightly anesthetized female can be visualized by phase contrast microscopy through the distended abdomen, after the intersegmental membranes are stretched by ventral placement of a glass cover slip. Most females recovered after the procedure were capable of subsequent reproductive activities. Albeit tedious, this technique preserves the female alive for subsequent experiments or observations. Its extension to other mosquito species, or other Diptera and insects, will depend on spermathecal and sperm visibility through the distended abdomen.

Association Between the Presence of Sperm in the Vasal Fluid During Vasectomy Reversal and Postoperative Patency: A Systematic Review and Meta-analysis

To investigate the association between the presence of sperm in the vasal fluid during vasectomy reversal (VR) and postoperative patency. We performed a systematic review and meta-analysis of the English-language literature reporting on the association between the presence of sperm in the intraoperative vasal fluid (ie, whole or parts vs none) and patency (ie, patent or not) after microsurgical vasovasostomy for men with obstructive azoospermia due to vasectomy. Odds ratios (OR) and 95% confidence intervals were calculated to quantify the strength of the association reported by each study. Meta-analysis was performed using a random-effects model. Four case series and 2 retrospective cohort studies of a total of 1293 eligible patients were identified. The mean age at VR was 37.8years, and the mean obstructive interval was 7.1years. The unadjusted OR of postoperative patency was 4.1 times higher (95% confidence interval, 2.3-7.3) given the presence of intravasal sperm or sperm parts as opposed to their absence at the time of VR (Q= 3.4; df= 5; P= .6; I(2)= 22%). The pooled OR should be interpreted with caution as only the 2 retrospective cohort studies reported meaningful data on this association. Because of inconsistent reporting, analysis of other vasal fluid characteristics (eg, consistency) and outcomes (eg, pregnancy) was not possible. The presence of whole sperm or sperm parts in the vasal fluid during VR is positively associated with postoperative patency. Our review highlights the poor methodological quality of existing evidence and underscores the need for more thorough follow up and higher standards of reporting in future studies.

Intra‐ and intermale variability of mature sperm traits analysed in two brackish water populations of the pipefish Syngnathus abaster (Syngnathidae)

AbstractSperm cells are highly diversified in animals, and considerable research effort has focused on variation in sperm morphology among species. Surprisingly, little is known about intraspecific variation in sperm morphology. We analysed within‐ and between‐male variation in mature sperm traits in two brackish water populations of the pipefish Syngnathus abaster. Four morphometric parameters, such as the width and length of the head (including nucleus, and midpiece), length of flagellum and total sperm length were taken into account. The differences in all morphometric parameters analysed between populations were not statistically significant. Moreover, the multidimensional scaling analysis shows that (i) the two populations seem to be indistinguishable based on their spermatozoa and (ii) there is not polymorphism, being sperm not distinguishable into discrete classes both within a single male and between males of each populations. The latter datum does not seem to support the presence of polymorphic sperm in syngnathids. Both populations, however, exhibit a high variation in all sperm traits, both among individual sperm within an ejaculate and among males within each population. The relationship between sperm traits variability and the low selection pressure determined by the absence of postcopulatory sexual selection (i.e. absence of sperm competition) is discussed.

Impact on Pregnancy of Gross and Microscopic Vasal Fluid during Vasectomy Reversal

We compared fertility outcomes with gross and microscopic fluid findings at vasectomy reversal at a high volume vasectomy reversal center. A retrospective study of a prospective database was performed. All vasectomy reversals were performed by a single surgeon (EFF) between 1978 and 2011. The clinical pregnancy rate was self-reported or determined via patient mailers. Patient and operative findings were determined through database review. We classified vasal fluid as opalescent, creamy, pasty or clear. Intraoperative light microscopy was used to determine if sperm or sperm parts were present and if they were motile. Multivariate analysis was performed evaluating patient age, partner age, years after vasectomy, type of surgery, and gross and microscopic fluid analysis. A total of 2,947 microsurgical vasectomy reversals were reviewed after we excluded reversals performed for post-vasectomy pain. We determined the pregnancy status of 902 (31%) cases. On univariate analysis with respect to pregnancy the presence of motile sperm at vasovasostomy neared statistical significance (p=0.075) and there was no difference between bilateral vs unilateral motile sperm. Gross fluid appearance was not statistically significant but we found the order of pregnancy success to be opalescent, creamy, clear then pasty fluid. On multivariate analysis only female partner age and sperm heads only or no sperm seen on light microscopy had statistical significance (p <0.05). The presence of motile sperm at vasectomy reversal approaches statistical significance on univariate analysis as a factor that affects clinical pregnancy rates. On multivariate analysis female partner age and microscopic findings of sperm heads only or no sperm are inversely related to pregnancy rates. These data will help counsel couples after vasectomy reversal and reinforce the importance of female partner age.

Microscopic Analysis of Mustelus schmitti´s Uteri During Reproductive Cycle

Aims: It was analysed with light and electron microscopy the different zones of the Mustelus schmitti`suteri along the reproductive cycle. Study Design: Histological routine techniques were used. Place and Duration of Study: Specimens of Mustelus schmitti were collected from the Bahia Blanca estuary, each month, from 2002 to 2006. They were caught by longlines. The fishing was made at the top of the tide during the day. Methologogy: A total of 24 animals were collected and processed; 11 were immature, 6 were maturing and 7 mature females; 4 of this last group were pregnant. Each animal was humanely sacrificed. A longitudinal ventral incision was made in each animal so as to remove the reproductive organs. The pieces were stored in Bouin solution for light microscopy and in 2.5% glutaraldehyde in 0.1 M cacodylate with 12% saccarose for transmission. Results: In immature animals, the simple columnar epithelium shows small and numerous folds, PAS (-) and AB (-). Beneath this epithelium, muscular fibers in longitudinal disposition alternate with connective tissue, finishing in a simple serosa. The uterine epithelium of the anterior portion in mature females, between the isthmus and the cervix, is stratified, PAS (+) and AB (+). Three muscle layers continue and the first one displays a disposition like a net. The second layer has longitudinal fibers divided into two by vascularized connective tissue and the third, is very thin. A typical serosa encloses the organ. In the posterior Original Research Article Elias; ARRB, 6(1): 20-32, 2015; Article no.ARRB.2015.060 21 portion of the uterus in mature females, the epithelium is also stratified but with more secretive cells. The presence of sperm was detected in the cervix and in the posterior portion of the uterus suggesting a special uterine epithelial-sperm interaction. Conclusion: The uteri suffer a cyclic remodelation not only in the epithelia but in the muscle layers.