Obesity causes weight increases in prepubertal and pubertal male offspring and is related to changes in spermatogenesis and sperm production in rats.

The effect of obesity on testicular activity in prepubertal and pubertal rats was investigated in the present study. Obesity was induced in adult females by feeding a high-calorie diet (HCD). These females were mated with normal males and were fed an HCD during pregnancy and lactation. The male offspring born to obese mothers and fed an HCD after weaning were found to be obese. Seminiferous tubules of offspring from control mothers (OCM) and offspring from HCD-fed mothers (OHCDM) had the same set of germ cells at different age intervals, namely spermatogonia, leptotene spermatocytes, zygotene spermatocytes, pachytene spermatocytes and round and elongated spermatids on postnatal days (PND) 7, 13, 17, 24 and 36, and on the day of preputial separation, respectively. However, there was a significant decrease in round and elongated spermatids and the epididymal sperm count, coupled with a significant decrease in testosterone and an increase in leptin serum concentrations in OHCDM compared with OCM. These results show that obesity in prepubertal rats does not affect the age-dependent appearance of germ cells according to developmental hierarchy, but it does interfere with spermatid formation, resulting in a reduced sperm count, which may be due to a deficiency of testosterone mediated by hyperleptinaemia.

Effects of caudal sufentanil supplemented with levobupivacaine on blocking spermatic cord traction response in pediatric orchidopexy.

PURPOSES: Caudal block is one of the most commonly used anesthetic techniques in subumbilical and genitourinary procedures. However, traditional administration of caudal levobupivacaine was inadequate on blocking peritoneal response during spermatic cord traction. The aim of this study was to evaluate whether the addition of caudal sufentanil to levobupivacaine provided better analgesia for children undergoing orchidopexy. METHODS: Sixty-two patients, scheduled for right orchidopexy, received caudal block after induction. Group LS (n = 31) received levobupivacaine 0.25% 1 ml/kg plus sufentanil 0.5 µg/kg, and group L (n = 31) received levobupivacaine 0.25% 1 ml/kg only. HR or MAP fluctuation >20% or entropy increase >15% during spermatic cord traction was defined as inadequate anesthesia and was treated with increasing sevoflurane concentration. The number of children who needed sevoflurane rescue was counted, and postoperative side effects and quality of sleep were also recorded. RESULTS: There were no statistically significant differences between the two groups in age, weight, and duration of surgery. Two (6.45%) children in group LS required inspired sevoflurane rescue to block hemodynamic fluctuation during spermatic cord traction, as compared with 12 (38.71%) patients in group L (P < 0.001). At the time of exerting spermatic cord traction, the median HR was, respectively, 134 and 145 (P < 0.001); the corresponding response entropy (RE) and state entropy (SE) was 65 and 54, respectively, in group LS versus 76 and 65 in group L (P < 0.001). CONCLUSION: In pediatric orchidopexy, the addition of sufentanil to levobupivacaine for caudal blockade offers clinical benefit over levobupivacaine alone in blocking the spermatic cord traction response.

Effects of compression/stretching of the spermatic cord and blunt dissection on testicular growth and fertility.

PURPOSE: This study was performed to investigate whether compression/stretching of the spermatic cord or blunt dissection influences testicular development and fertility. In addition, the authors evaluated whether the extents of testicular damage differ between these 2 surgical manipulations. METHODS: Forty-four prepubertal male Sprague-Dawley (SD) rats (Harlan Sprague-Dawley Inc, Indianapolis, Ind) were divided into 3 groups: (1) the control group (CG) animals underwent a sham operation in the right groin, (2) the experimental group 1 (EG1) underwent compression/stretching of the right spermatic cord, and (3) the experimental group 2 (EG2) underwent dissection around the right spermatic cord structures. Testicular volumes, weights, mean seminiferous tubular diameters (MSTDs), mean testicular biopsy scores, and numbers of offspring and of pregnant females were evaluated. RESULTS: Right (operative) and left (nonoperative) testicular volumes were smaller in the EG2 group than in the CG or EG1 groups. Left MSTDs in the EG1 and EG2 groups increased more than in the CG group. Numbers of Sertoli cells in left testes differed in the 3 groups, in the order EG1 < CG < EG2. Mean testicular biopsy scores, offspring numbers, and pregnant female numbers were no different in the 3 groups. CONCLUSIONS: Both surgical manipulations influenced testicular growth, but they did not compromise spermatogenesis or fertility in SD rats.

Pulsed radiofrequency for chronic testicular pain-a preliminary report.

OBJECTIVE: To evaluate the effectiveness of pulsed radiofrequency (PRF) of spermatic cord in the treatment of chronic testicular pain. DESIGN: Ten patients with chronic testicular pain were treated with PRF stimulation of the spermatic cord. A radiofrequency probe placed percutaneously into the spermatic cord was used to deliver four 120-second cycles of 20-millisecond pulses at 2 Hz. Test stimulation was first used to confirm the precise placement of the probe. The short-form McGill Pain Questionnaire was used to assess pain before treatment and at 3 months. Patients who had experienced improvement were followed up by telephone, to determine whether pain relief was sustained. RESULTS: Ten patients were entered into the study but one was lost to follow-up. Of the nine patients evaluated, four had complete resolution of pain, while one had partial pain relief. Three patients experienced no change and one reported that his pain was worse. All patients who experienced complete and partial pain relief continued to do so at a mean long-term follow-up of 9.6 months (range 3-14 months). There were no complications observed immediately or during the follow-up period. CONCLUSION: In this pilot study, pain scores improved in five out of nine patients. PRF of spermatic cord appears to be a safe minimally invasive outpatient procedure that should be investigated further with placebo-controlled trials.

Identification and characterization of Myosin from rat testicular peritubular myoid cells.

In the mammalian testis, peritubular myoid cells (PMCs) surround seminiferous tubules. These cells are contractile, express the cytoskeletal markers of true smooth muscle-alpha-isoactin and F-actin-and participate in the contraction of seminiferous tubules during the transport of spermatozoa and testicular fluid to the rete testis. Myosin from PMCs (PMC-myosin) was isolated from adult rat testis and purified by cycles of assembly-disassembly and sucrose gradient centrifugation. PMC-myosin was recognized by a monoclonal anti-smooth muscle myosin antibody, and the peptide sequence shared partial homology with rat smooth muscle myosin-II, MYH11 (also known as SMM-II). Most PMC-myosin (95%) was soluble in the PMC cytosol, and purified PMC-myosin did not assemble into filaments in the in vitro salt dialysis assay at 4 degrees C, but did at 20 degrees C. PMC-myosin filaments are stable to ionic strength to the same degree as gizzard MYH11 filaments, but PMC-myosin filaments were more unstable in the presence of ATP. When PMCs were induced to contract by endothelin 1, a fraction of the PMC-myosin was found to be involved in the contraction. From these results we infer that PMCs express an isoform of smooth muscle myosin-II that is characterized by solubility at physiological ionic strength, a requirement for high temperature to assemble into filaments in vitro, and instability at low ATP concentrations. PMC-myosin is part of the PMC contraction apparatus when PMCs are stimulated with endothelin 1.

A real-time dynamic 3D model of the human inguinal region for surgical education.

A suitable dynamic 3D model that allows the simulation of the inguinal region with real-time performance on a personal computer was developed. A geometric model adjusted to real data was created by means of semiautomatic contour segmentation of anatomic units from the visible human project and data generated from classical anatomic information. A dynamic model included converting muscular units from their continuous geometric representation into a set of voxels and then real-time interaction and performance. The current implementation enables deformation of the realistic model associated with pushing and stretching interaction, allowing immersion in the anatomy of the inguinal structures. The model does not allow simulation of surgical interventions.

Study of the cremasteric muscle during erection.

BACKGROUND: Mechanism of testicular elevation during erection is not known. We investigated the hypothesis that erection evokes reflex cremasteric muscle (CM) contraction which effects testicular elevation. METHODS: Electromyographic (EMG) response of CM to erection was recorded in 26 healthy volunteers (age 36.7 +/- 6.8 SD years). Erection was induced by intracavernosal injection of alprostadil. CM response was tested before and after individual glans penis (GP) and CM anesthetization. RESULTS: The CM exhibited resting electric activity of mean amplitude of 74.8 +/- 6.3 microV which, on erection, increased to 486.6 +/- 36.8 microV (P < 0.001). Response was momentary. Anesthetization of erect GP did not effect increase of CM EMG activity, while bland gel did. Anesthetized CM did not respond to GC erection while saline infiltrated did. CONCLUSIONS: The CM appears to contract during erection through a reflex which we call 'peno-cremasteric reflex'. CM contraction assumingly elevates testicle and support cord veins; it may effect testicular compression, thus expressing its secretions into vas deferens.

Straining-cremasteric reflex: identification of a new reflex and its role during increased intra-abdominal pressure.

The cremasteric muscle (CM) being composed of fleshy muscle bundles constitutes the active component of the fasciomuscular tube of the spermatic cord. On contraction, the CM compresses the cord veins pushing the blood in the pampiniform plexus to the abdominal veins. The role of the CM during increased intra-abdominal pressure (IAP) could not be traced in the literature. We investigated the hypothesis that the CM contracts upon IAP increase so as to support the cord veins and prevent abdominal veins reflux into them. Thirty-two healthy male volunteers (mean age 40.2 +/- 11.2 SD years) were studied. The IAP was recorded by a manometric catheter introduced into the rectum. The CM response to straining (sudden by coughing and slow by Valsalva's maneuver) was registered by a needle electrode introduced into the muscle. The response was recorded again after individual anesthetization of the CM and rectum. The test was repeated using saline instead of lidocaine and was performed on both sides. Straining (sudden or slow sustained) effected increase of the rectal pressure and CM EMG. The more the rectal pressure was increased by straining, the more the CM EMG was increased. The CM EMG response disappeared after prolonged or repeated successive straining. The CM did not respond to straining after individual anesthetization of the rectum and CM but did respond to saline administration. The response was similar from muscles on both sides. Increased CM EMG on straining postulates a reflex relationship which we call the 'straining-cremasteric reflex' (SCR). We suggest that this reflex, which results in CM contraction, supports the spermatic cord veins against the increase of the IAP induced by straining and against the tendency of venous reflux from the abdominal veins. The SCR may prove of diagnostic significance in neurogenic disorders provided further studies are performed in this respect.

The effect of spermatic vessel ligation on testicular nitric oxide levels and germ cell-specific apoptosis in rat testis.

Management of high testis may vary but the most popular method in surgical treatment is the Fowler-Stephens maneuver. The aim of the present study was to investigate the effects of spermatic vessel ligation on testicular nitric oxide (NO) levels, expression of inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS) and germ cell-specific apoptosis in both ipsilateral and contralateral testes in rats. Twenty-eight animals were randomly allocated into four groups (n=7 each). The spermatic vessels were ligated as a simulation of the Fowler-Stephens maneuver. The groups of animals were sacrificed at 2 h (group 1), 4 h (group 2) and 24 h (group 3) after ligation, respectively. Sham-operated animals served as controls (group 4). Biochemical assessment of testicular NO levels was performed by the Griess method. iNOS and eNOS expression and apoptosis were studied in ipsilateral and contralateral testes. Testicular NO levels at 24 h after the simulated Fowler-Stephens maneuver were found to be significantly increased in both ipsilateral and contralateral testes when compared with the sham-operated group. eNOS expression was clearly increased in ipsilateral testes, whereas moderate expression was detected in the contralateral seminiferous tubules at 24 h after ligation. Mild focal iNOS immunostaining was also observed in seminiferous tubules of the ipsilateral testis at 24 h after the simulated Fowler-Stephens maneuver. Apoptosis was dramatically increased in ipsilateral testes; however, it was only detected in single cells in the contralateral side at 24 h after ligation. In conclusion, the simulated Fowler-Stephens maneuver induces testicular nitric oxide synthesis and germ cell-specific apoptosis in the ipsilateral testis. These results suggest that high levels of NO induce apoptosis and may impair spermatogenesis thus explaining the unsuccessful outcome of the Fowler-Stephens maneuver.

An intravital microscopy study of radiation-induced changes in permeability and leukocyte-endothelial cell interactions in the microvessels of the rat pia mater and cremaster muscle.

Using intravital microscopy and a closed window method, we measured irradiation-induced changes in the vascular permeability and cell interactions in microcirculation networks of the rat pia mater; the same effects were monitored in the cremaster muscle as a control. The closed cranial window has many advantages, including long-term direct visualization of microcirculation. The method allows for repeated testing of the same vessel or network, thereby reducing variability. The method also allows for measurement of permeability changes and the accompanying leukocyte-endothelial cell interactions in the same network or vessel, which permits correlative studies of these phenomena. However, this method is not without challenges. The optical conditions are difficult, because the brain is three-dimensional and its parenchyma is more complex than the thinner, flatter peripheral tissues. To overcome this limitation, we performed a dynamic background subtraction. The background is dynamically related to vessel intensity, and changes in intensity were determined by eliminating the effects of neighboring and underlying vessels. We applied this method to studying the effects of ionizing radiation on the blood-brain barrier (BBB) permeability and cell interactions and the modulation of these effects by anti-ICAM-1 antibodies. Our results demonstrate that this method is sensitive to changes in these properties of the BBB.

Three-dimensional geometrical models of the inguinal region. Towards a new stereology.

In this work we studied the inguinal-abdominal region and the inguinal canal using three-dimensional geometrical models. We built the models through computer aided geometric modeling techniques on the basis of observations during real dissections, operations and diagnostic medical imaging. The obtained models show in a complete modular synthesis and with a schematic iconology the structural organization of the anatomical districts in a logic sequence of layers and topographic and spatial relationships among its components. The models represent an amazing support to anatomy and clinical anatomy for teaching and research purposes on organogenesis, surgery and diagnosis.

Caudal anesthesia in children: effect of volume versus concentration of bupivacaine on blocking spermatic cord traction response during orchidopexy.

UNLABELLED: In this study we compared the intensity and level of caudal blockade when two different volumes and concentrations of a fixed dose of bupivacaine were used. Fifty children, 1-6 yr old, undergoing unilateral orchidopexy received a caudal block with a fixed 2 mg/kg dose of bupivacaine immediately after the induction. Group 1 (n = 23) received 0.8 mL/kg of 0.25% bupivacaine, whereas Group 2 (n = 27) received 1.0 mL/kg of 0.2% bupivacaine. Epinephrine 1:400,000 and 0.1 mL of sodium bicarbonate per 10 mL of local anesthetic solution were added. There were no statistically significant differences between the two groups in their anesthesia, surgery, recovery, and discharge times. Fifteen patients (65.2%) in Group 1 required an increase in inspired halothane concentration to block hemodynamic and/or ventilatory response during spermatic cord traction, as compared with 8 patients (29.6%) in Group 2 (P = 0.022). In the recovery room, four (17.4%) patients in Group 1 required rescue treatment with fentanyl, versus two (7.4%) in Group 2 (P = 0.372). In children undergoing orchidopexy, a caudal block with a larger volume of dilute bupivacaine is more effective than a smaller volume of the standard 0.25% solution in blocking the peritoneal response during spermatic cord traction, with no change in the quality of postoperative analgesia. IMPLICATIONS: In children undergoing orchidopexy, a caudal block with a larger volume of dilute bupivacaine is more effective than a smaller volume of the more concentrated solution in blocking the peritoneal response during spermatic cord traction, with no change in the quality of postoperative analgesia.

Excitation and sensitization of the heat response induced by a phorbol ester in canine visceral polymodal receptors studied in vitro.

To clarify the possible involvement of protein kinase (PK) C activation in the excitation and sensitization of polymodal receptors (PMRs), the effects of phorbol 12,13-dibutyrate (PDBu) on PMRs were studied in canine testis-spermatic nerve preparations in vitro. Application of PDBu (10(-7), 10(-6), AND 10(-5) M) for 5 min evoked a significant increase in the ongoing activity of the PMRs within 15 min. PDBu (10(-8) to 10(-5) M) significantly augmented the subsequent heat responses of the PMRs. Staurosporine (10(-6) M), a PK inhibitor, attenuated the effect of PDBu on heat responses. These data suggest that activation of PKC contributes to the activities of PMRs.

Vascular morphology of the golden hamster spermatic cord.

The morphology of the convoluted testicular artery and the pampiniform plexus of the golden hamster was studied by light microscopy and corrosion cast techniques combined with scanning electron microscopy. The artery was found to be totally enclosed by the pampiniform plexus, except for minor superficial areas where the artery was exposed. Although no direct connection between the artery and the vein was found in the area of apposition, the arterial and venous walls reduced their thickness by sharing a single tunica adventitia, which seemed well suited to the transfer of substances by diffusion. Many band-like structures of the venous walls were found in the deep part of the spermatic cord, suggesting that these may act as barriers to slow down the venous blood velocity. The venous wall here and there showed a stick-like endothelial bridge, suggesting that it may prevent the veins from over distension. In addition to the close relation between the artery and the vein, lymphatic vessels and mast cells were distributed widely within the connective tissue of the arterio-venous walls and venous walls. Mast cells were situated mainly in the area of apposition, especially at the base of the protruding venous wall. These morphological findings suggest that mast cells may be involved in the counter-current transfer mechanism in the spermatic cord of the golden hamster.

Cooling prevents ischaemic testicular damage after spermatic cord clamping in rats.

A controlled trial was conducted on four groups of seven rats to evaluate any beneficial effect of cooling to 15 degrees C on testicular ischaemia induced by clamping of the spermatic cord for 2, 4, 6 or 8 h. At operation the microscopic appearance of the testes on unclamping was a poor guide to ultimate viability, whereas by the third day macroscopic and microscopic appearances concurred. When killed on the third day, testicular histology demonstrated that increasing periods of clamping led to gross ischaemia and infarction when testes were maintained at 37 degrees C, and this was reduced greatly by testicular cooling. For each period of clamping, maintenance of spermatogenesis, expressed in terms of Johnsen's histological scores, was significant (P less than 0.01), being 2 (37 degrees C) vs 5.3 (15 degrees C) for 8-h clamping. It is proposed that testicular cooling should be used during autotransplantation of testes in humans.

Sperm duct contractions mediate centrally evoked sperm release in goldfish.

In order to determine the peripheral mechanisms underlying sperm release (SR) in goldfish, the contractile activity of the sperm ducts (SD) and testes were monitored during SR responses evoked by electrical stimulation of the brain. Electrical stimulation of the brain triggered testicular and SD contractions, and SR, while electrical stimulation of the genital nerve branch to the SD evoked only SD contractions and SR. Centrally activated SD contractions and SR were blocked by sectioning the SD genital nerve, while testicular contractions were unaffected. Testicular contractions do not appear necessary for centrally evoked SR since the response can be elicited from preparations in which the testes were separated from the SD. The results indicate that SR in goldfish is primarily mediated by the SD and not the testes. Testicular contractions may, however, serve to load the SD with milt. The functional significance of the central pathway(s) associated with SD and testicular contractions are discussed.

Morphological and physiological factors involved in the contractility of the spermatic cord and ductus deferens of the opossum (Didelphis albiventris).

The opossum spermatic cord is characterized by a thick skeletal muscle coat, provided by the great development of the cremasteric muscle. Acetylcholine induced a powerful contraction of the spermatic cord placed in an organ bath containing Heller's solution. This contraction was blocked by curare but not completely by atropine as is usual for skeletal muscle. However, the ductus deferens did not respond to acetylcholine or catecholamines under the same conditions. Specific histochemical methods for both catecholamines and acetylcholinesterases revealed that the ductus deferens musculature, composed mainly of circular fibres, is richly innervated by adrenergic and presumed cholinergic nerve fibres. The lack of responsiveness to the neurotransmitters could be explained by the absence of longitudinally arranged muscle. It is suggested that in the opossum the cremaster could participate in the mechanism of sperm transport.