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Prognostic factors for relapse in stage I seminoma managed by surveillance: a pooled analysis.

PURPOSE: Several management options are available to patients with stage I seminoma, including adjuvant radiotherapy, surveillance, and adjuvant chemotherapy. We performed a pooled analysis of patients from the four largest surveillance studies to better delineate prognostic factors associated with disease progression. PATIENTS AND METHODS: Individual patient data were obtained from each center (Princess Margaret Hospital, Danish Testicular Cancer Study Group, Royal Marsden Hospital, and Royal London Hospital) for 638 patients. Tumor characteristics (size, histologic subtype, invasion of rete testis, and tumor invasion into small vessels [SVI]) as well as age at diagnosis were analyzed for prognostic importance for relapse. RESULTS: With a median follow-up of 7.0 years (range, 0.02 to 17.5 years), 121 relapses were observed for an actuarial 5-year relapse-free rate (RFR) of 82.3%. On univariate analysis, tumor size (RFR: 4 cm, 76%; P =.003), rete testis invasion (RFR: 86% [absent] v 77% [present], P =.003), and the presence of SVI (RFR: 86% [absent] v 77% [present], P =.038) were predictive of relapse. On multivariate analysis, tumor size ( 4 cm, hazard ratio 2.0; 95% confidence interval [CI], 1.3 to 3.2) and invasion of the rete testis (hazard ratio 1.7; 95% CI, 1.1 to 2.6) remained as important predictors for relapse. CONCLUSION: We have identified size of primary tumor and rete testis invasion as important prognostic factors for relapse in patients with stage I seminoma managed with surveillance. This information will allow patients and clinicians to choose management based on a more accurate assessment of an individual patient's risk of relapse. In addition, it will allow clinicians to tailor follow-up protocols based on risk of occult disease.

Adult↗

[Lymphatic system of the human testis].

The distribution of lymph vessels in the human testis was investigated using ink injection methods, light and electron microscopy. Lymph capillaries occur in the septula testis but are absent in the intertubular tissue. They consist of endothelial cells provided with an incomplete basal lamina and bound by anchoring filaments to the adjacent connective tissue. Frequently, the endothelial cells are separated by gaps (measuring up to 2 microns). The lymph capillaries of the septula testis are connected to lymph vessels in the rete testis and tunica albuginea. These vessels have occasional smooth muscle cells and valves. At the posterior margin of the testis, the network of lymphatics merges into collecting ducts that together with vessels derived from the rete testis are drained by lymphatics in the spermatic cord.

Adolescent↗

Effect of ultrasound on testicular electrolytes (sodium and potassium).

The testes of 50 rats were placed in a cup filled with water and received 1 W/cm2 of ultrasound for 15 min. Fluid was collected from the seminiferous tubules and rete testis of the treated and control groups at 1, 8, 12, and 24 hr intervals. Ultrasound increased the sodium concentration in the fluid of the seminiferous tubules, decreased the sodium concentration in the fluid of the rete testis, increased the potassium concentration in the fluid of the rete testis, and decreased the potassium concentration in the fluid of the seminiferous tubules. Fourteen, slightly sedated, monkeys (Macaca fascicularis) were treated with 1/2 W/cm2 of ultrasound for 30 min. Water was used as the coupling agent for seven monkeys and 3% NaCl was used as the coupling agent for the other seven monkeys. The efficacy of ultrasound treatment in reducing sperm count to zero and achieving zero motility was increased when 3% NaCl was used. Sperm count was at the level of presonication after 20 weeks when water was used as a coupling agent.

Animals↗

Distribution of keratins, vimentin, and actin in the testis of two South American camelids: vicuna (Vicugna vicugna) and llama (Lama glama). An immunohistochemical study.

The purpose of the present study was to investigate the pattern of distribution of cytokeratins, vimentin and muscular actin in the testis of vicuna (Vicugna vicugna) and llama (Lama glama) two species of camelids native of the Andean high plateau of South America. Testicular biopsies of four vicunas and five llamas were used. Animals were healthy breeders. The tissues were processed by standard immunohistochemistry with antipancytokeratinAE1/AE3, antikeratin 18 (K 18), CAM 5.2 (antikeratin 5, 18, and 19), antivimentin, and smooth-muscle-specific antiactin antibodies to track the cytoskeletal pattern of testicular cells. Using AE1/AE3 antibody the immunostaining was found in the epithelial lining of tubuli recti and rete testis. The reaction was relatively stronger in the apical cytoplasm of epithelial cells. The testicular cells of the two species showed no reaction to K 18 and CAM 5.2 antibodies. Antivimentin antibody stained the basal cytoplasm of the Sertoli cells, the Leydig cells, and the epithelial lining of tubuli recti and rete testis. In the last two structures the immunostain was relatively more intense in the basal cytoplasm of epithelial cells. Antiactin antibody stained the peritubular cells and the muscle cells of the lamina propria oftubuli recti and rete testis. The presence in these species of only some keratins found in man, its coexpression with vimentin in epithelial lining of tubuli recti and rete testis and the peritubule organization, so different from other ungulates may reflect a differential adaptation of the cytoskeleton to particular reproductive strategies.

Actins↗

Degeneration of the seminiferous epithelium with ageing is a cause of spermatoceles?

A spermatocele refers to the cystic accumulation of semen in the male reproductive tract. Although it is thought to be caused by narrowing of the lumen of the excurrent duct with resultant cystic dilatation of the duct, the pathogenesis of the narrowing remains unknown. In the present study, we histologically examined spontaneous spermatoceles in C3H/He mice to elucidate the pathogenesis of the lesions. Testes, efferent ducts, epididymides and vas deferens obtained from young and aged C3H/He mice were embedded in plastic for histological observation at the light microscopic level. It was found that spontaneous spermatoceles were localized in the rete testis and efferent ducts of aged mice, as seen in man. The dilated rete testis and efferent ducts contained many degenerated and aggregated germ cells derived from the exfoliated seminiferous epithelium in the aged testis. In particular, it was noted that the agglutinated germ cells obstructed the narrow lumen of the efferent ducts, resulting in the failure of transport of germ cells to the caput epididymis, and spermatoceles were consistently found in the region between the rete testis and the obstructed site in the efferent ducts. However, no inflammatory cell infiltration, traumatic injury or spermatic granulomas were found in the occluded region. These results suggest that agglutinated germ cells may occupy the narrow lumen of the efferent ducts, resulting in the formation of a spermatocele. It may be that a senile change to the seminiferous epithelium, which releases immature germ cells into the lumen of the seminiferous tubules, is the cause of this type of spermatocele.

Aging↗

Histogenetic consideration of ovarian sex cord-stromal tumors analyzed by expression pattern of cytokeratins, vimentin, and laminin. Correlation studies with human gonads.

A total of 30 sex cord-stromal tumors including 9 adult type and 5 juvenile type granulosa cell tumors (GCTs), 4 Sertoli-Leydig cell tumors (SLTs), 1 gynandroblastoma, 5 thecomas, 2 fibromas and 3 sclerosing stromal tumors were immunohistochemically evaluated by means of cytokeratins of different molecular weight, vimentin and laminin with regard to the histogenesis of these tumors and to the embryogenesis of the sex cord and stroma of developing gonads. For comparison, 7 embryonic gonads, 9 fetal and 9 adult ovaries, 14 fetal and 5 postnatal testes, and 1 gonadoblastoma were also examined. The coelomic epithelium of all gonads were positive for both cytokeratins (CAM 5.2 and AE1) and vimentin. In fetal ovaries, the granulosa cells of primordial follicles express low molecular weight cytokeratins only and those cells of more maturing follicles did not express any cytokeratin or vimentin. In adult ovaries, the granulosa cells of primordial follicles coexpressed low molecular weight cytokeratins and vimentin, but those cells of more maturing follicles expressed vimentin only. In fetal testes before 20 weeks gestational age, the Sertoli and Leydig cells did not express any cytokeratins and vimentin. After that time, both cells expressed vimentin only throughout life. The rete ovarii and rete testis from fetal to adult life coexpressed both low molecular weight cytokeratins and vimentin. The rete ovarii in all ages and rete testis in prenatal and childhood ages were surrounded by the laminin-positive basement membrane, however, the rete testis in adult were not. In neoplasia, the GCTs, thecomas, fibromas, and sclerosing stromal tumors expressed vimentin only.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Intratesticular spermatocele.

High resolution ultrasound has increased our awareness of benign testicular cysts that appear to arise from the intratesticular sperm conduit system of the intratesticular ducts of the mediastinum and rete testis. The location of these benign cysts in the area of the mediastinum and rete testis and the absence of signs of a germ cell tumor may permit testicle preservation. These benign cysts may contain sperm. We present a case of a benign sperm-containing cyst with visible communication to the rete testis, designated an intratesticular spermatocele and managed with testicle preservation.

Adult↗

Expression of constitutively active Notch1 in male genital tracts results in ectopic growth and blockage of efferent ducts, epididymal hyperplasia and sterility.

The Notch signaling pathway is involved in a variety of developmental processes. Here, we characterize the phenotypes developing in the reproductive organs of male transgenic (Tg) mice constitutively expressing the activated mouse Notch1 intracellular domain (Notch1(intra)) under the regulatory control of the mouse mammary tumor virus (MMTV) long terminal repeat (LTR). Tg expression was detected in testis, vas deferens and epididymis by Northern blot analysis. In situ hybridization with a Notch1-specific probe lacked sensitivity to detect expression in normal-appearing cells, but demonstrated expression in hyperplastic epithelial cells of the vas deferens, epididymis and efferent ducts. Tg males from three independent founder lines were sterile. Histological analysis of reproductive organs of young Tg males (postnatal ages 8 and 21) showed no difference compared to those of non-Tg males. In contrast, in adult Tg mice from day 38 onwards, the efferent ducts, the vas deferens and most epididymal segments revealed bilateral epithelial cell hyperplasia with absence of fully differentiated epithelial cells. Electron microscopy confirmed the uniformly undifferentiated state of these cells. Immunohistochemistry with anti-PCNA antibody also revealed enhanced proliferation of Tg epididymis. In adult Tg testis, the different generations of germ cells of seminiferous tubules appeared normal, although some tubules were highly dilated and revealed an absence of early and/or late spermatids. The epithelial cells of the Tg tubuli recti and rete testis were not abnormal, but the rete testis was highly dilated and contained numerous spermatozoa, suggesting a downstream blockage. Consistent with a blockage of efferent ducts often seen at the rete testis/efferent duct interface, spermatozoa were absent in epididymis of all adult Tg mice and in all highly hyperplastic efferent duct tubules of these Tg mice. Such a blockage was visualized by injection of Evans blue dye into the rete testis lumen. Finally, the presence of ectopic hyperplastic efferent duct tubules was observed within the testicular parenchyma itself, outside their normal territory, suggesting that Notch1 signaling is involved in the establishment of these borders. This phenotype seems to represent a novel developmental defect in mammals. Together, these results show that constitutive Notch1 signaling significantly affects the development of male reproductive organs.

Animals↗

Role of epithelial cells of the male excurrent duct system of the rat in the endocytosis or secretion of sulfated glycoprotein-2 (clusterin).

The localization of sulfated glycoprotein-2 (clusterin; SGP-2) was investigated in the rete testis, efferent ducts, and epididymis of the rat using light (LM) and electron (EM) microscope immunocytochemistry. At the LM level, the epithelial cells of the rete testis and efferent ducts demonstrated an intense immunoperoxidase reaction over their apical and supranuclear regions, and sperm in the lumen of the efferent ducts were unreactive. In the EM, gold particles were found exclusively over the endocytic apparatus of these cells. In the proximal area of the epididymal initial segment, an insignificant immunostaining of epithelial cells and sperm was observed. However, the distal area of the initial segment showed a moderate staining over the epithelial principal cells and sperm, while in the intermediate zone of the epididymis a stronger reaction was observed over these cells. The strongest immunoperoxidase reaction was noted in the caput epididymidis, where it formed a distinct mottled pattern. Thus, while some principal cells were intensely stained, others were moderately or weakly stained; a few were completely unreactive. In the corpus and cauda epididymidis, the staining pattern was similar but not as intense. In the EM, only the secretory apparatus of these cells was found to be immunolabeled with gold particles. Sperm in the lumen of these different regions were also labeled. The epithelial clear cells were unreactive throughout the epididymis. Northern blot analysis substantiated these results and showed the presence of highest levels of SGP-2 mRNA in the caput epididymidis, especially in its proximal area, whereas increasingly lower levels were found in the corpus and cauda epididymidis. In summary, these results suggest that testicular SGP-2 dissociates from the sperm during passage through the rete testis and efferent ducts, where it is endocytosed by the epithelial cells lining these regions. In the epididymis, it is replaced by an epididymal SGP-2 that is secreted by the epithelial principal cells of the epididymis. Furthermore, in the epididymis, the principal cells appear to be in different functional states with respect to the secretion of epididymal SGP-2 within a given region of the duct as well as along the epididymal duct.

Animals↗

Distribution pattern of F-actin, vimentin and alpha-tubulin in the bovine testis during postnatal development.

The distribution of F-actin, vimentin and alpha-tubulin was studied immunohistochemically in bovine seminiferous and straight testicular tubules, rete testis and intertubular tissue during postnatal development. Sites of antigenicity were detected by ABC immunoperoxidase technique and visualized by metal-enhanced deposition of diaminobenzidine. Within the seminiferous epithelium, F-actin appears at 20 weeks and is found in adult Sertoli cells as part of specialized cell contacts. In peritubular cells, F-actin increases gradually from 4 to 30 weeks when the adult concentration is achieved. After 20 weeks, subepithelial fibroblasts of the mediastinum testis start to express F-actin and at 52 weeks, a thick layer of positive myofibroblasts is seen beneath the epithelia of rete testis and straight testicular tubules. Testicular macrophages and light intercalated cells (LIC) are also characteristically decorated following F-actin immunoreaction. Vimentin is localized in perinuclear position in pre-Sertoli cells of 4-20 weeks and in adult Sertoli cells. During the period of transformation from pre-Sertoli to Sertoli cells, the perinuclear vimentin coat is absent. The epithelia of rete testis and straight tubules exhibit a strong vimentin immunoreaction in their basal parts. This specific pattern does not change from 4 weeks to adulthood. Alpha -tubulin is absent in 4-week-old seminiferous tubules. At 8 weeks, the perinuclear area of pre-Sertoli cells reacts positive. The alpha-tubulin content increases in these cells continuously, and from 30 weeks on nearly the entire supranuclear cytoplasm of Sertoli cells is heavily decorated. The epithelial of rete and straight tubules display a growing number of alpha-tubulin-positive cells from 4 to 40 weeks. From then on, nearly all epithelial cells contain alpha-tubulin, particularly in a narrow zone beneath their lateral cell borders.

Actins↗

Secretion of D-aspartic acid by the rat testis and its role in endocrinology of the testis and spermatogenesis.

The D-isomer of aspartic acid (D-Asp) has been found in rat testes. In the present study, samples of testicular venous blood plasma, rete testis fluid, interstitial extracellular fluid, luminal fluid from the seminiferous tubules, testicular parenchymal cells, epididymal spermatozoa and peripheral blood plasma were collected and analyzed for D-Asp by two methods, an enzymatic and a chromatographic HPLC method. The two methods gave very similar results for all samples. The highest concentrations of D-Asp (about 120 nmol/ml) were found in testicular venous blood plasma, with slightly lower concentrations in rete testis fluid (95 nmol/ml) and epididymal spermatozoa (80 nmol/g wet weight). Lower levels were found in testicular parenchymal cells (which would comprise mostly spermatids and spermatocytes), luminal fluid from the seminiferous tubules and interstitial extracellular fluid (26, 23 and 11 nmol/ml respectively). However, these values were all higher than those for peripheral blood plasma (6 nmol/ml). It would appear that D-Asp is being secreted by the testis mostly into the venous blood, passing thence into the rete testis fluid and being incorporated into the spermatozoa at the time or after they leave the testis. The distribution of D-Asp is thus quite different from that of testosterone, and its role and the reason for its high concentration in the male reproductive tract remain to be elucidated.

Animals↗

Isolation and transplantation of spermatogonia in sheep.

Studies in rodents show that spermatogonial transplantation is an excellent new tool for studying spermatogenesis and for preservation and dissemination of genetics. The aim of this study was to adapt the technique to rams. Two issues were addressed: purification of stem cell spermatogonia, and efficient injection of donor spermatogonia into the seminiferous tubules of rams. We compared differential plating and Percoll gradient methods for purifying donor spermatogonia from ram lamb testes. Spermatogonia were identified with an antibody against PGP 9.5, a ubiquitin C-terminal hydrolase. Both purity and total number of spermatogonia recovered were higher after purification by Percoll gradient than by differential plating. Four approaches for injecting cells into the seminiferous tubules of ram testes were compared ex vivo: insertion of a needle into the extra-testicular rete testis after reflection of the head of the epididymis ('surgical' approach), and ultrasound-guided insertion of a needle into the extra-testicular rete, and the proximal and distal parts of the intra-testicular rete testis. 'Surgical' and ultrasound-guided approaches into the extra-testicular rete resulted in highest success rates and best filling of the seminiferous tubules. Finally, the ultrasound guided approach into the extra-testicular rete testis was validated in vivo by transplanting purified spermatogonia previously labeled with a fluorescent molecule (CFDA-SE). In seven of eight testes injected, donor cells were identified within the seminiferous epithelium for up to 2wk after transplantation, indicating the integration of donor cells.

Animals↗

LGR4 regulates the postnatal development and integrity of male reproductive tracts in mice.

The roles of the leucine-rich repeat domain containing G protein-coupled receptor (GPCR) 4 (Lgr4), which is one of the orphan GPCRs, were analyzed with the Lgr4 hypomorphic mutant mouse line (Lgr4(Gt)). This homozygous mutant had only one-tenth the normal transcription level; furthermore, 60% of them survived to adulthood. The homozygous male was infertile, showing morphologic abnormalities in both the testes and the epididymides. In the testes, luminal swelling, loss of germinal epithelium in the seminiferous tubules, and rete testis dilation were observed. Cauda epididymidis sperm were immotile. Rete testis dilation was due to a water reabsorption failure caused by a decreased expression of an estrogen receptor (ESR1) and SLC9A3 in the efferent ducts. Although we found differential regulation of ESR1 expression in the efferent ducts and the epididymis, the role of ESR1 in the epididymis remains unclear. The epididymis contained short and dilated tubules and completely lacked its initial segment. In the caput region, we observed multilamination and distortion of the basement membranes (BMs) with an accumulation of laminin. Rupture of swollen epididymal ducts was observed, leading to an invasion of macrophages into the lumen. Male infertility was probably due to the combination of a developmental defect of the epididymis and the rupture of the epithelium resulting in the immotile spermatozoa. These results indicate that Lgr4 has pivotal roles to play in the regulation of ESR1 expression, the control of duct elongation through BM remodeling, and the regional differentiation of the caput epididymidis.

Animals↗

Immunolocalisation of oestrogen receptor-alpha within the testis and excurrent ducts of the rat and marmoset monkey from perinatal life to adulthood.

The sites of action and the physiological role of oestrogens in the male reproductive tract are poorly understood. We have undertaken a systematic study of the immunoexpression of oestrogen receptor-alpha (ER alpha) in the male rat from late fetal life through to adulthood and compared the findings with results obtained in the marmoset monkey (Callithrix jacchus) from neonatal to adult life. The testes, rete testis, efferent ducts and epididymis were examined from normal male rats (aged 4, 8, 10, 15, 20, 25, 38, 48 and 90 days) and from male rat fetuses on days 17.5 and 18.5 of gestation; comparable tissues were examined from neonatal, infantile, peripubertal and adult marmosets aged 8, 18-24, 54-62 and 92-112 weeks respectively. Immunolocalisation of ER alpha used antigen retrieval and a monoclonal antibody directed to the N-terminus, which had proved superior to six other antisera tested. ER alpha was immunoexpressed in interstitial cells, including the fetal/ neonatal generation of Leydig cells, in both the rat and marmoset. In the rat, the adult generation of Leydig cells were also immunopositive for ER alpha whereas the comparable cells in the marmoset were only weakly immunopositive. ER alpha was not expressed in Sertoli cells, peritubular myoid cells, blood vessels or germ cells at any time in either species. In late fetal life in the rat, ER alpha was immunoexpressed in cells surrounding the mesonephric tubules, whereas postnatally it was expressed in the epithelium of the rete testis and efferent ducts at all ages from 4 to 90 days; this immunoexpression was most pronounced in the efferent ducts. In the marmoset, the efferent ducts, but not the rete testis, also showed intense immunoexpression of ER alpha. Apart from sporadic immunostaining for ER alpha in the epididymal duct of the rat in the neonatal period, the caput, corpus and cauda epididymis were negative for immunoexpression of ER alpha at all ages in both species. These findings suggest that the main actions of oestrogens in the male reproductive tract, mediated by ER alpha, are related to the development and function of the efferent ducts and the Leydig cells. In consideration of data from this and previous studies of oestrogen binding, we predict possible sites of expression of other oestrogen receptors (e.g. ER beta) in Sertoli cells and the epididymis. Interactive effects, related to the relative levels of androgens and oestrogens, could be physiologically important in the excurrent ducts of the adult testis.

Age Factors↗

A new approach to study the architectural arrangement of spermatogenic stages revealed little evidence of a partial wave along the length of human seminiferous tubules.

Although evidence of the spermatogenic wave in humans has been reported, little is known about the architectural arrangement of stages along the length of tubules in men with varied rates of sperm production. To conduct these studies, methodology is needed to stage relatively long lengths of straight portions of tubules in several men. Testes obtained at autopsy were glutaraldehyde perfused. Six men representing a range of daily sperm production were selected for this study. Wedges composed of groups of seminiferous tubules attached to the rete testis and separated by testicular septa were teased apart from the remaining testicular tissue, fixed in osmium, and oriented during Epon embedding. This orientation allowed for sections to be cut perpendicular to the length of the group of tubules, beginning at the end near where the tubules had been attached to the rete testis. Also, a tubule isolated away from the rete testis was oriented for perpendicular sectioning for one of the men. Several serial sections were cut at a setting of 22 microns, and each was optically sectioned approximately at 7 microns and photographed using Nomarski optics. Because many cytoplasmic and nuclear features of spermatogonia, spermatocytes, and spermatids were distinguished in photomicrographs produced by Nomarski optics, this optical system facilitated the identification of stages of the spermatogenic cycle in human seminiferous tubules.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗

Ontogeny of anti-müllerian hormone, 3 beta-hydroxysteroid dehydrogenase and androgen receptor expression during ovine total gonadal development.

Anti-müllerian hormone (AMH) and androgenic steroids are key factors regulating the masculinisation of the internal and external genitalia during fetal development. AMH is produced in Sertoli cells and causes regression of the müllerian ducts in the male. 3 beta-Hydroxysteroid dehydrogenase (3 beta-HSD) is one of the key steroidogenic enzymes regulating testosterone production in Leydig cells. The objective of this experiment was to elucidate the development of the ovine fetal testes by identifying the spatio-temporal expression of AMH, 3 beta-HSD and androgen receptor expression within them. Fetuses from days 30 and 40 of gestation were fixed intact, while the gonads were dissected from the fetuses on days 70, 100 and 130 of gestation. Tissue was fixed in Bouin's fixative for 6 h, processed into paraffin wax and sections immunostained using rabbit anti-human AMH, 3 beta-HSD or androgen receptor antibodies. While seminiferous cords were absent on day 30 of gestation, pre-cord organisation was apparent and the gonad could be clearly distinguished from surrounding tissue by the presence of AMH and 3 beta-HSD immunopositive cells. Androgen receptor expression was not apparent at this stage. By day 40 of gestation the testis was organised into distinct seminiferous cords and intense immunostaining for AMH and 3 beta-HSD was present in Sertoli cells within the cords and Leydig cells in the interstitium respectively. Androgen receptor immunopositive cells were present in the interstititum but cells destined to develop into rete testis were immunonegative. By day 70 of gestation, the rete testis was organised in the centre of the testis and was strongly androgen receptor immunopositive. AMH and 3 beta-HSD expression was present in Sertoli and Leydig cells respectively. The expression of AMH, 3 beta-HSD and androgen receptor in the 100 and 130 day gestation fetuses was similar to that identified in the 70 day fetuses. In conclusion, Sertoli and Leydig precursor cells are present in the gonad prior to seminiferous cord formation and contain AMH and 3 beta-HSD at all stages of gestation examined. While androgen receptor immunoexpression was present in nuclei of interstitial cells from day 40 of gestation and in the rete testis from day 70 of gestation, Sertoli cells were immunonegative for androgen receptor at all of the stages examined.

3-Hydroxysteroid Dehydrogenases↗

Microassay of sperm concentration in the rat epididymis by micropuncture technique.

Micropuncture samples taken from the rete testis, caput, corpus and cauda epididymidis of adult rats were assayed for intraluminal sperm concentrations. The amount of fluid resorbed from the efferent duct and epididymal lumen was calculated based on the sperm concentration. Epididymal sperm concentrations increased from the rete fluid to the cauda fluid through the caput fluid. Eighty-nine percent of the fluid leaving the rete testis is resorbed by the efferent ducts and proximal epididymal tubule, and 96% of the fluid leaving the rete testis is resorbed in the distal cauda epididymidis. Resorption is important for the control of intraluminal fluid by the epididymis.

Animals↗

Unusual observations on a serologically negative bluetongue virus infected bull.

A Holstein bull named "Regency" born in New York in July 1968, maintained in an artificial insemination (AI) stud for 9 years, unremarkable from the point of view of health history or semen production, was discovered "by accident" to have bluetongue (BT) virus (BTV) in his semen. A microscopic study in 1975 and 1977 revealed unusual cytoplasmic vacuoles in the sperm of "Regency" prompting us to send semen to A. J. Luedke, USDA-ARS, Denver, Colorado, USA, to attempt virus isolation. Serotype 13 BTV was isolated from the semen. Serum sent with the semen was negative for antibody by the agar gel immunodiffusion (AGID) and complement fixation (CF) tests. Annual AGID serologic tests begun in 1975 and continuing to the present have been negative for antibody to BTV for all bulls in this AI stud. "Regency" was moved to Auburn, Alabama, USA in 1978 and further studies were performed. The bull was maintained with the Veterinary School herd. The BT serologic status of the cattle did not change during the year the bull was in the herd nor were the cows he bred or their calves serologically positive for BT. Studies on insemination of cattle with experimentally contaminated semen indicated that 5X10(4) infectious doses of BTV were required for infection. These results suggest that since "Regency's" semen did not infect cows, the semen probably contained less than this amount of virus. The rete testis was cannulated, rete fluid collected, a testis removed and virus isolation attempted.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗