Evidence of a Wave of Spermatogenesis in Human Testis*

Hinweis auf das Vorliegen einer Welle der Spermatogenese im menschlichen Hoden In Samenkanälchen von zwei alten Männern war eine ungewohnt klare Gliederung des Keimepithels in Stadien erkennbar. An flächenhaften Rekonstruktionen der Wand von Samenkanälchen konnte die Existenz wendelförmig angeordneter Stadiensequenzen festgestellt werden. Ähnliche Befunde ließen sich auch bei einem jüngeren Patienten erheben. Die vorliegenden Ergebnisse weisen darauf hin, daß es auch im menschlichen Samenkanälchen eine Welle der Spermatogenese gibt. Es ist denkbar, daß die besondere lokale Organisation des Keimepithels beim Menschen auf einem spezifischen Modus der Spermatogoniogenese beruht.     

The Connection between the Seminiferous Tubules and the Rete Testis in the Domestic Fowl (Gallus Domesticus) Morphological Study

The tubules connecting the seminiferous tubules proper to the rete testis in the fowl were studied with the aid of light and electron microscopy. The material examined ultrastructurally was fixed by vascular perfusion through the thoracic aorta. The seminiferous tubules were joined to the rete testis in three different ways; they were either linked by a terminal segment and a tubulus rectus, by a terminal segment only, or opened directly into the rete cavities. The terminal segment of the seminiferous tubules was lined with columnar cells (modified Sertoli cells). These cells were characterized by having an indented nucleus with a prominent nucleolus, many mitochondria, a sizable Golgi apparatus, electron dense bodies and many cytoplasmic protrusions into the lumen. Intraepithelial lymphocytes as well as macrophages in the lumina of the terminal segment, the tubuli recti and the rete testis were also observed. Myoid cells were found in the boundary tissue of the terminal segment.     

Stages of the Cycle of the Seminiferous Epithelium in the Dog

Stadieneinteilung der normalen Spermatogenese beim Hund
Prinzipiell weist der Hund eine ähnliche Spermatogenese wie andere Mammalia auf. Er besitzt 6 aufeinanderfolgende, differenzierende Spermatogoniengenerationen (A1–A4, Im, B). Die Anzahl der nachfolgenden Zellgenerationen der Spermatogenese ent-spricht den Verhältnissen bei den übrigen Mammalia.
Die Spermatogenese des Hundes wird in 10 Stadien eingeteilt. In den Stadien II, III, V, VI, VII, VIII finden die Teilungen der hintereinandergeschalteten Spermato-gonien statt. Stadien V a und V c sind durch die Reifeteilungen gekennzeichnet.     

氟桂利嗪对小鼠生精细胞T型Ca~(2+)通道的作用

目的:研究二苯烷基氨类衍生物氟桂利嗪(F lu)对小鼠生精细胞T型钙电流(ICaT)的影响。方法:采用急性机械分离的小鼠生精细胞,应用膜片钳全细胞记录技术,观察F lu对ICaT的影响。结果:钳制电位为-90 mV、刺激电压为-30 mV时,F lu(0.1、1、10、100μmol/L)可抑制小鼠生精细胞Ca2+电流,抑制率分别为(23.34±2.76)%、(46.04±3.52)%、(62.52±3.70)%、(73.52±3.12)%(P<0.05);F lu对T型Ca2+通道的半数最大抑制浓度为0.29μmol/L。结论:F lu对生精细胞T型Ca2+通道有阻断作用,呈浓度依赖性;Cav3.2是生精细胞T型Ca2+电流的主要贡献者。     

胱蛋白酶抑制剂相关的附睾精子发生基因在不同发育阶段小鼠睾丸和附睾中的表达

目的 :研究胱蛋白酶抑制剂相关的附睾精子发生 (Cres)基因在生后不同发育阶段小鼠睾丸及附睾中的表达规律。　方法 :采用半定量RT PCR方法检测CresmRNA在生后 14、2 0、2 2、2 8、35、4 9、70、4 0 0d小鼠睾丸及附睾中的表达变化。　结果 :Cres基因在 14d小鼠睾丸和附睾中呈低水平表达 ,随着小鼠的生长发育 ,CresmRNA的表达量逐渐升高。在 70d小鼠睾丸和 4 0 0d小鼠附睾中 ,CresmRNA的表达量达到峰值。　结论 :Cres基因在生后不同发育阶段小鼠睾丸及附睾中呈现明显的规律性表达 ,可能参与精子发生、成熟过程的调控。     

Lepidium meyenii （Maca） reduces spermatogenic damage induced by a single dose of malathion in mice

AIM: To observe the effect of the aqueous extract of hypocotyls of the plant Lepidium meyenii (Maca) on spermatogenic damage induced by the organophosphate insecticide malathion in mice. METHODS: Mice were treated with 80 mg/kg of malathion in the presence or absence of an aqueous extract of Maca, which was orally administered 7, 14 or 21 days after injection of the malathion. Stages of the seminiferous epithelium were assessed by transillumination on days 0, 7, 14 and 21. RESULTS: The administration of Maca increased significantly the length of stage VIII on days 7, 14 and 21 of treatment compared with the controls. An increase in the length of stage IX occurred on day 14 of treatment. Malathion affected spermatogenesis by reducing the lengths of stage IX on day 7, stages VII and IX-XI on day 14 and a recovery of stages IX-XII on day 21. The magnitude of alteration in the length of stage IX produced by malathion was significantly reduced by Maca on days 7 and 14. The length of stage VIII was increased when Maca was administered to mice treated with malathion. Assessment of the relative length of stages of the seminiferous epithelium showed that Maca treatment resulted in rapid recovery of the effect of malathion. CONCLUSION: Maca enhances spermatogenesis following spermatogenic damage caused by the organophosphorous pesticide.     

Radical inguinal orchidectomy: the gold standard for initial management of testicular cancer

Radical inguinal orchidectomy with division of the spermatic cord at the internal inguinal ring is the gold standard for diagnosis and local treatment of testicular malignancies. The technique is well established and described in detail in this paper, collating methods from various surgical textbooks and articles. We also discuss pre-operative considerations including fertility counselling and potential testicular prosthesis at time of orchidectomy, and the importance of contemplating differential diagnoses such as para-testicular sarcoma and primary testicular lymphoma (PTL) prior to performing radical orchidectomy (RO). The evidence and indications for new surgical techniques to treat local testicular malignancies are also described, including testis sparing surgery (TSS) and spermatic cord sparing orchidectomy.     

Comparative immunolocalisation of perlecan, heparan sulphate, fibroblast growth factor-18, and fibroblast growth factor receptor-3 and their prospective roles in chondrogenic and osteogenic development of the human foetal spine

Purpose

A comparative immunolocalisation study of perlecan, HS, FGF-18 and FGFR-3 in the 12–20-week gestational age human foetal spine was undertaken to identify spatiotemporal associations between these components to provide insights into prospective roles in spinal development.

Methods

Comparative immunolocalisations of matrix and cell associated components in Histochoice-fixed paraffin-embedded human foetal spinal tissues.

Results

The 12–14-week-old human foetal spine was a predominantly cartilaginous structure with the discs displaying a relative paucity of proteoglycan compared to the adjacent cartilaginous vertebral rudiments, notochordal remnants were also observed. HS and perlecan had a widespread distribution throughout the spine at 12 weeks, however, FGF-18 was only localised to the outer AF margins and hypertrophic cell condensations in the vertebral bodies. This contrasted with HS distributions at 14–20 weeks, which were prominent in the developing intervertebral disc (IVD). Ossification centres were also evident centrally within the vertebral rudiments surrounded by small columns of hypertrophic chondrocytes which expressed FGFR-3 and FGF-18 and upregulated levels of perlecan. FGF-18 also had a prominent localisation pattern in the developing IVD and the cartilaginous endplate while FGFR-3 was expressed throughout the disc interspace. This suggested roles for perlecan, FGF-18 and FGFR-3 in chondrogenic and osteogenic events which drive discal development and ossification of the vertebral bodies.

Conclusions

The above data supported a role for FGF-18 in discal development and in the terminal osteogenic differentiation of chondroprogenitor cell populations, which promote vertebral ossification during spinal development.     

Genetic analysis of the human Insulin‐like 3 gene: absence of mutations in a Greek paediatric cohort with testicular maldescent

This study investigated the hypothesis that genetic alterations of the human insulin‐like 3 (INSL3) gene are associated with testicular maldescent (TMD). Genomic DNA was extracted and amplified from peripheral blood samples of 170 unrelated children with all possible phenotypical expressions of TMD and 50 volunteers with normal external genitalia from the general paediatric population (controls). PCR‐single strand conformation polymorphism analysis was used to screen INSL3 gene for genetic variants. For rapid screening of a detected nonsilent genetic alteration, restriction assay using endonuclease Eag I was further employed. Products were analysed on 2% agarose gel and restriction patterns were visualised by ethidium bromide. Differences in genotype and allelic distributions of nonsilent genetic alterations were evaluated between (i) patients–controls, (ii) familial–sporadic, (iii) bilateral–unilateral and (iv) intra‐abdominal–inguinal cases of TMD. No mutations were detected. Three common INSL3 gene polymorphisms (27G>A, 126G>A, 178G>A) unrelated to any particular phenotype of TMD were detected both in patients and controls. These results indicate that INSL3 gene mutations are not a common cause of TMD in the human.     

Comparison of sperm preparation methods: effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer

The aim of this study was to evaluate the efficacy of swim-up, PureSperm gradient centrifugation and glass-wool filtration methods for semen preparation and to assess the possible enhancement of the quality of the subpopulation of spermatozoa in terms of sperm concentration, morphology and chromatin condensation. Moreover, to determine the effect of this semen processing technique on the clinical outcome after in vitro fertilization embryo transfer (IVF-ET). A total of 180 semen samples of patients' husbands who were undergoing IVF therapy were prepared by swim-up (G1, n = 60), PureSperm gradient centrifugation (G2, n=60) or glass-wool (G3, n=60) methods. Chromatin condensation was assessed by Chromomycin (CMA3), whereas sperm morphology was evaluated according to strict criteria. In all three semen processing methods, the percentage of chromatin condensed and morphologically normal spermatozoa was higher after semen processing in comparison with native semen samples. The proportion of normal chromatin condensed spermatozoa prepared in glass-wool filtration was significantly higher than that in swim-up (G.I, p=0.02) or PureSperm (G.II, p=0.001). In addition semen processing with PureSperm yields significantly a higher percentage of morphologically normal spermatozoa than swim-up (p < 0.001) or glass-wool method (p < 0.002). However, the fertilization, implantation and pregnancy rates, in turn were similar in all semen preparation methods. In conclusion, PureSperm gradient centrifugation yields a higher percentage of morphologically normal spermatozoa than shown in traditional swim-up or glass-wool filtration. However, the percentage of chromatin condensed spermatozoa was significantly higher after semen processing via glass-wool in comparison with the other two methods. Nevertheless, there were no significant difference in the fertilization, implantation and pregnancy rates of sperm prepared by means of swim-up, PureSperm or glass-wool filtration. Therefore, glass-wool filtration should be recommended as the first choice for semen preparation for Intracytoplasmic sperm injection (ICSI) technique as the natural selection is bypassed. Whereas, swim-up and PureSperm should be used for semen processing in IVF programme.     

Prostate stem cell antigen (PSCA) mRNA expression in prostatic intraepithelial neoplasia: implications for the development of prostate cancer

BACKGROUND: Prior data clearly demonstrated the expression of prostate stem cell antigen (PSCA) mRNA in prostatic intraepithelial neoplasia (PIN) tissues. The purpose of the present investigation was to determine whether PSCA mRNA expression was associated with the presence of cancer in this disease. METHODS: One hundred seventeen men were diagnosed with isolated PIN on initial prostate biopsy, 51 with low-grade form (LGPIN), and 66 with high-grade form (HGPIN). PSCA mRNA expression in initial PIN and subsequent cancer was examined by in situ hybridization (ISH). The differences of the PSCA mRNA expression level between the groups were analyzed by the Chi-square and Student's t-test. Univariate and multivariate logistic regression analyses were performed to evaluate the predictive performance of PSCA mRNA. RESULTS: PSCA mRNA expression level in 34 subsequent cancers was statistically increased compared with their paired PIN (P < 0.001), with a Gleason's dependence. HGPIN showed statistically high PSCA mRNA expression compared with LGPIN (P < 0.01). PSCA mRNA expression levels were significantly stronger in the initial isolated LGPIN and isolated HGPIN with subsequent cancer than those without (P < 0.001 and P < 0.001, respectively). Multivariate logistic regression analysis demonstrated that only PSCA mRNA was predictive of the onset of subsequent cancer in patients with isolated LGPIN and in those with isolated HGPIN, respectively. CONCLUSIONS: Our data identify PSCA mRNA in initial PIN as a significant predictor of subsequent cancer, suggesting that PSCA implies in prostatic tumorigenesis and may be used to identify the patients with isolated PIN who are at high risk for cancer onset in the disease process.     

Determinants of seeking of primary care for lower urinary tract symptoms: the Krimpen study in community-dwelling men

OBJECTIVES: To determine which factors predict seeking of primary care among men with lower urinary tract symptoms (LUTS). METHODS: A longitudinal, population-based study with a follow-up period of 6.5 yr was conducted among 1688 men aged 50-78 yr. The following data were collected: prostate volume (using transrectal ultrasonometry), urinary flow rate, ultrasound-estimated postvoid residual urine volume, generic and disease-specific quality of life (QOL), and symptom severity (International Prostate Symptom Score [IPSS]). Information on the seeking of primary care by all participants during 2 yr of follow-up was collected from the general practitioner's (GP) record using a computerised search engine and a manual check of electronically selected files. RESULTS: Prostate volume, postvoid residual volume, IPSS, and social generic QOL are important determinants of first GP consultation in men with LUTS. Measurements (physical urologic parameters) and self-reported items (symptom severity and QOL) contribute almost equally to GP consultation in these men. CONCLUSIONS: Both measurements of prostate volume and postvoid residual urine volume and self-reported information about symptoms or QOL can help to select those who will benefit from medical care and to reassure those men not likely to need help in the near future.     

In vivo studies comparing the biocompatibility of various polypropylene meshes and their handling properties during endoscopic total extraperitoneal (TEP) patchplasty: an experimental study in pigs

Background: Polypropylene (PP) meshes are currently being used in millions of operations for hernial repair. We tested several recently modified PP meshes to evaluate them in terms of their biocompatibility and handling characteristics during minimally invasive procedures. Methods: Four different PP meshes (a heavyweight PP, Atrium; a lightweight PP incorporating absorbable polyglactic acid, Vypro II; a lightweight PP, Parietene; and a titanium-coated lightweight PP, Ti-Mesh Extralight), all implanted using the endoscopic total extrapreperitoneal (TEP) technique, were investigated in groups comprising 11 pigs each. After 94 ± 5 days, the animals were autopsied and tissue samples were studied histologically, immunohistochemically, and electron-microscopically. Results: Whereas endoscopic handling of the Vypro II mesh proved difficult, the already good properties of the Atrium mesh were significantly improved on by Parietene and the Ti-Mesh. Clear differences were also found in the shrinkage characteristics of the implant materials. In comparison with Atrium (12%), Vypro II mesh shrinkage was significantly greater (28%), whereas Parietene (7%) and Ti-Mesh (5%) incurred less shrinkage. With regard to the chronic inflammatory reaction, the titanium-coated mesh showed a significantly lower inflammatory activity (13.1% partial volume [%PV] vs 34.1%PV and 29.0%PV) than the lightweight meshes Vypro II and Parietene, but—with the exception of the monocytic (0.2 vs 9.1, 5.1, and 7.9) and B-lymphocytic reaction (1.1 vs 18.0, 11.7, and 12.2)—no significant difference was seen in comparison with Atrium. The various mediators in the extracellular matrix (matrix metalloproteinases 1 [MMP-1 transforming growth factor beta [TGF-], urokinase plasminogen activator [uPA], and type I collagen) tended to show the highest expression with Vypro II (13.6, 113.2, 132.7, and 139.5, respectively) and the lowest expression with Ti-Mesh (11.9, 68.5, 92.8, and 75.0, respectively). With regard to cell proliferation, Parietene and Ti-Mesh appeared to have slight advantages, but no differences were observed in the apoptotic rate. Conclusion: In our opinion, despite a reduction in material, Vipro II, on account of the inflammatory reaction, does not represent a true improvement over the meshes currently used for hernia repair. In comparison, the two lightweight PP patches are characterized by a more favorable foreign body reaction, with the titanium coating of the Ti-Mesh providing an additional advantage in terms of its biocompatibility.     

A transverse rectus abdominis myocutaneous (TRAM) flap for reconstruction of a large deformity in the buttock: A new indication for a TRAM flap

A rectus abdominis myocutaneous flap has been used to repair the abdominal wall, chest wall, sternum, breast, and groin. We describe a patient in whom a large deformity in the buttock caused by a road crash was repaired with a transverse rectus abdominis myocutaneous (TRAM) flap. Distally-based TRAM flaps provide a good and reliable way of reconstructing the buttock. Its advantages are a long arc of rotation and well-vascularised bulky tissue that serve as a partition and promote quick healing of the defect. However, the flap is not the first choice for traumatic and infected wounds where fatty tissue is not desired.     

Modulation of the C1 visual event-related component by conditioned stimuli: evidence for sensory plasticity in early affective perception

Previous research has demonstrated optimized processing of motivationally significant stimuli early in perception. In the present study, the time course and underlying mechanisms for such fast differentiation are of interest. We investigated the involvement of the primary visual cortex in affective evaluation of conditioned stimuli (CSs). In order to elicit learning within the visual system we chose affective pictures as unconditioned stimuli and used laterally presented gratings as CSs. Using high-density electroencephalography, we demonstrated modulation of the C1 visual event-related component for threat-related stimuli versus neutral stimuli, which increased with continuing acquisition of affective meaning. The differentiation between aversive and neutral visual stimuli occurred as early as 65-90 ms after stimulus onset and suggested involvement of the primary visual areas in affective evaluation. As an underlying mechanism, we discuss short-term reorganization in visual cortex, enabling sensory amplification of specific visual features that are related to motivationally relevant information.