Perforant path activation of ectopic granule cells that are born after pilocarpine-induced seizures.

Granule cells in the dentate gyrus are born throughout life, and various stimuli can affect their development in the adult brain. Following seizures, for instance, neurogenesis increases greatly, and some new cells migrate to abnormal (ectopic) locations, such as the hilus. Previous electrophysiological studies of this population have shown that they have intrinsic properties that are similar to normal granule cells, but differ in other characteristics, consistent with abnormal integration into host circuitry. To characterize the response of ectopic hilar granule cells to perforant path stimulation, intracellular recordings were made in hippocampal slices from rats that had pilocarpine-induced status epilepticus and subsequent spontaneous recurrent seizures. Comparisons were made with granule cells located in the granule cell layer of both pilocarpine- and saline-treated animals. In addition, a few ectopic hilar granule cells were sampled from saline-treated rats. Remarkably, hilar granule cells displayed robust responses, even when their dendrites were not present within the molecular layer, where perforant path axons normally terminate. The evoked responses of hilar granule cells were similar in several ways to those of normally positioned granule cells, but there were some differences. For example, there was an unusually long latency to onset of responses evoked in many hilar granule cells, especially those without molecular layer dendrites. Presumably this is due to polysynaptic activation by the perforant path. These results indicate that synaptic reorganization after seizures can lead to robust activation of newly born hilar granule cells by the perforant path, even when their dendrites are not in the terminal field of the perforant path. Additionally, the fact that these cells can be found in normal tissue and develop similar synaptic responses, suggests that seizures, while not necessary for their formation, strongly promote their generation and the development of associated circuits, potentially contributing to a lowered seizure threshold.

Comparison of microscopic methods for detecting inflammation in expressed prostatic secretions.

PURPOSE: We evaluated microscopic methods of detecting inflammation in expressed prostatic secretions. MATERIALS AND METHODS: Methods of counting expressed prostatic secretion leukocytes were compared in 251 samples from 159 patients with chronic prostatitis/chronic pelvic pain syndrome, including traditional wet mounts, hemocytometer derived concentrations and expressed prostatic secretion smears stained with Gram's method or DiffQuick stain (Dade International, Inc., Miami, Florida). RESULTS: Of 159 initial patient evaluations 84 (53%) showed inflammation by hemocytometer concentration at 500 leukocytes per mm.3 or greater but only 37 (23%) were considered inflammation by the traditional wet mount method (p <0.001). Inflammation was identified in 149 of 251 specimens (59%) by hemocytometer but in only 82 (33%) by wet mount (p <0.001). When inflammation was defined as 1,000 leukocytes per mm.3 or greater the hemocytometer still identified significantly more patients (41%) and specimens (48%) with inflammation than the wet mount. The hemocytometer method had a substantially lower interassay and intra-assay coefficient of variation than the wet mount method. Polymorphonuclear neutrophils and macrophages were the most common cells observed on stained smears, which detected inflammation in 147 specimens (59%) by DiffQuick but in only 98 (39%) by Gram's method. CONCLUSIONS: Detecting inflammation in expressed prostatic secretions is method dependent. Significantly more cases of inflammation were detected by hemocytometer than by the traditional wet mount technique. Because the wet mount method also proved more variable than the hemocytometer and highly sensitive to volume, its use is not recommended. These findings support the adoption of hemocytometer and staining methods for accurate evaluation of expressed prostatic secretion inflammation in men with chronic prostatitis/chronic pelvic pain syndrome.

Neurophysiologic testing in chronic pelvic pain syndrome: a pilot study.

OBJECTIVES: Studies of male chronic pelvic pain syndrome (CPPS) have generally centered on the pathologic features of the prostate rather than on the neurology of pain. Electrodiagnostic studies examine the integrity of somatosensory nerve pathways consisting of large, group A fibers. Heat sensation and visceral (autonomic) pain is mediated through small, unmyelinated C fibers, which can be tested cutaneously by thermal sensory analysis. We hypothesized that CPPS pain is mediated by these small C fibers. METHODS: All subjects and controls had no history of neurologic disease and had normal neurologic examinations. Phase I: 14 patients with CPPS underwent electrodiagnostic testing using pudendal somatosensory evoked potentials and bulbocavernosus reflex latency measurements. Phase II: 31 patients with CPPS and 14 controls underwent thermal sensory analysis testing on the perineum and anterior thigh using noxious heat stimuli. Subjects used a computer-generated visual analog scale to dynamically report their discomfort. The peak and slope of the computer-generated visual analog scale were analyzed. RESULTS: Phase I: two patients had delayed latency of the somatosensory evoked potentials, but additional evaluation with magnetic resonance imaging revealed no definable lesion. Phase II: with thermal sensory analysis, men with CPPS reported higher intensity pain at lower temperatures (P = 0.03). Men with CPPS also had higher peak computer-generated visual analog scale scores on perineal testing. No difference in thermal testing on the anterior thigh was found between the two groups. CONCLUSIONS: Large, myelinated somatic fibers do not play a significant role in the pathophysiology of CPPS. Patients with CPPS have an altered sensation of perineal pain elicited by heat, which may represent a C-fiber-mediated effect.

Consensus development of a histopathological classification system for chronic prostatic inflammation.

OBJECTIVE: To develop a standardized histopathological classification system for chronic prostatitis (standardized description of prostatic inflammatory infiltrates) based on a literature review, extensive prospective evaluations in two recognized prostatitis research centres and widespread consensus of international urological centres identified as having major expertise or interest in chronic prostatitis. METHODS: Relevant articles for review were identified by a Medline search undertaken by the Cochrane Review Group in Prostate Diseases and Urologic Malignancies, and cross-checking bibliographies of retrieved studies, reviews, book chapters and abstracts of the American Urological Association and International Prostatitis Collaborative Network Annual Meetings. Initial drafts were based on classification systems independently developed by the Prostatitis Research Centers at Queen's University in Canada and University of Washington in the USA. A collaborative draft was distributed to 20 urological/pathological clinical centres who participated in the North American Chronic Prostatitis Collaborative Research Network and First International Prostatitis Collaborative Network. A consensus classification system was then distributed to the participating panel for acceptance. RESULTS: The literature review identified a reasonably consistent description of inflammatory infiltrate locations and patterns that were further incorporated into the draft based on the Queen's University and University of Washington proposals. Eighteen (90%) of the identified Prostatitis Centers participated in the revision of the draft and the final consensus process. The final consensus document classifies prostatic inflammation according to its extent and grade/severity in each tissue compartment (location). Conclusion The consensus of the expert panel was that this classification system can be used in the evaluation of prostatic inflammation in prostate biopsies, transurethral resected prostate chips or prostatectomy specimens. A standardized accepted framework to describe histopathological prostate inflammation will prove useful in evaluating prostate disease.

Should vasectomy reversal be performed in men with older female partners?

PURPOSE: An assumption exists that men with older female partners who seek treatment of post-vasectomy infertility should undergo in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) rather than vasectomy reversal. Although several studies have reviewed ICSI success rates with advancing maternal age, to our knowledge none has compared them to outcomes for vasectomy reversal in men with older partners. MATERIALS AND METHODS: The records of all patients with ovulating partners older than 37 years who underwent vasectomy reversal from 1994 through 1998 were reviewed. Patients were contacted to establish pregnancy and birth rates. Costs of vasectomy reversal, testicular sperm extraction, IVF and ICSI were obtained from the financial office of our institution. RESULTS: A total of 29 patients underwent vasectomy reversal with a followup of 3 to 59 months (median 25). Median male age was 46 years (range 37 to 67) and median female age was 40 years (range 38 to 48). A total of 5 pregnancies and 4 live births were achieved. In the 23 patients followed for more than 1 year the pregnancy rate was 22% and live birth rate was 17%. Using this 17% birth rate at our $4,850 cost for vasectomy reversal the cost per newborn was $28,530. In comparison, using the 8% birth rate per cycle of ICSI for women older than 36 years at a cost of $8,315 for testicular sperm extraction and 1 cycle of IVF with ICSI, the cost per newborn was estimated at $103,940. CONCLUSIONS: Vasectomy reversal appears to be cost-effective to achieve fertility in men with ovulating partners older than 37 years.

Pain associated with testicular retraction treated with Gore-Tex external inguinal ring reconstruction.

We present our experience with three patients with chronic testicular pain due to retractile testes and propose a new operative solution to the problem. Three patients with chronic testicular pain associated with testicular retraction and relieved by pushing the testicle into the scrotum were identified. Full history and physical examinations were performed to rule out other causes of testicular pain. The patients underwent open inguinal exploration, aborted attempt at repair of an attenuated or obliterated external oblique aponeurosis, and construction of a neo-external inguinal ring with a Gore-Tex strip. The patients were reevaluated in the clinic postoperatively to determine change in physical examination and symptoms. All three patients had nonretractile testes on follow-up examination and reported improvement of their testicular pain. An attenuated or torn external oblique aponeurosis can result in a patulous external inguinal ring and painful retractile testicle. If traditional orchidopexy is insufficient to prevent severe retraction, reconstruction of the external inguinal ring with Gore-Tex mesh can correct the anatomical deficiency, reduce testicular retraction, and improve pain symptoms.

Prostate histopathology and the chronic prostatitis/chronic pelvic pain syndrome: a prospective biopsy study.

PURPOSE: The chronic prostatitis/chronic pelvic pain syndrome is a common clinical syndrome characterized by lower genitourinary tract symptoms, particularly pain in the perineum or genitalia, voiding symptoms, such as dysuria or frequency, and sexual dysfunction in the absence of uropathogens in the urine or prostatic secretions. The term prostatitis is based on the presumption that prostatic inflammation is important in the pathophysiology of this syndrome. To our knowledge there has been no systematic characterization of the degree and nature of inflammation in the prostate in symptomatic cases. MATERIALS AND METHODS: Prostate histopathology in 368 biopsies from 97 patients with the chronic prostatitis/chronic pelvic pain syndrome was characterized.. RESULTS: Prostatic inflammation was detected in only 33% of patients, including 29% with mild (less than 10 leukocytes per 1 mm. field) and 4% with moderate (between 10 and 200) or severe (more than 200) infiltrate. Of the 3 patients with moderate inflammation 1 had glandular, 1 periglandular and 3 multifocal or diffuse distribution of leukocytes in the interstitium. Some patients had more than 1 pattern of inflammation. CONCLUSIONS: The finding of moderate or severe inflammation in only 5% of 97 patients argues for the need to reevaluate current concepts of the pathophysiology of the chronic prostatitis/chronic pelvic pain syndrome.

Chronic, low-level (1.0 W/kg) exposure of mice prone to mammary cancer to 2450 MHz microwaves.

In a previous study (Frei et al., Bioelectromagnetics 19, 20-31, 1998), we showed that low-level (0.3 W/kg), long-term exposure of mice prone to mammary tumors to 2450 MHz radiofrequency (RF) radiation did not affect the incidence of mammary tumors, latency to tumor onset, tumor growth rate or animal survival when compared to sham-irradiated animals. In the current study, the specific absorption rate (SAR) was increased from 0.3 W/kg to 1.0 W/kg. The same biological end points were used. One hundred C3H/HeJ mice were exposed in circularly polarized waveguides for 78 weeks (20 h/day, 7 days/week) to continuous-wave, 2450 MHz RF radiation; 100 mice were sham-exposed. There was no significant difference between exposed and sham-exposed groups with respect to the incidence of palpated mammary tumors (sham-exposed = 30%; irradiated = 38%), latency to tumor onset (sham-exposed = 62.0 +/- 2.3 weeks; irradiated = 62.5 +/- 2.2 weeks) and rate of tumor growth. Histopathological evaluations revealed no significant difference in numbers of malignant, metastatic or benign neoplasms between the two groups. Thus long-term exposures of mice prone to mammary tumors to 2450 MHz RF radiation at SARs of 0.3 and 1.0 W/kg had no significant effects when compared to sham-irradiated animals.

Vasectomy reversal associated with increased reactive oxygen species production by seminal fluid leukocytes and sperm.

PURPOSE: Reactive oxygen species, which are primarily produced by leukocytes, are generally detrimental to sperm. High reactive oxygen species levels are found in men with abnormal sperm function. Since men often have poor sperm characteristics and infertility after vasectomy reversal, we compared reactive oxygen species in seminal cells of men after vasovasostomy to those of fertile men to determine if reactive oxygen species were elevated in the former group. MATERIALS AND METHODS: We studied semen samples of men with proved fertility (39) and those with previously proved fertility who had undergone vasectomy reversal (45). The presence of leukocytes was determined by Bryan-Leishman staining. Reactive oxygen species endogenous activity was monitored by luminol dependent chemiluminescence in washed cells, including all cells in the semen, and Percoll density gradient purified sperm. RESULTS: After vasovasostomy men had significantly lower sperm concentration, motility and computerized motility measurements than fertile men. Mean reactive oxygen species in washed seminal cells after vasovasostomy was 684 relative light units per second compared to 49 for fertile controls (p < 0.0001). Density gradient purified sperm had 53 and 0.64 relative light units per second, respectively (p < 0.0001). When men with leukocytospermia were excluded from analysis, differences between the groups remained, although 9 times more reactive oxygen species were detected in men after vasectomy reversal with than those without leukocytes in semen. CONCLUSIONS: Higher levels of reactive oxygen species are found in washed seminal cells and purified sperm after vasectomy reversal than in those of fertile men. Although leukocytes are probably a significant source of reactive oxygen species in these groups, they may not account for all of the increased reactive oxygen species after vasovasostomy. Low motility after vasectomy reversal may be related to the detrimental effects of reactive oxygen species produced by leukocytes or sperm, even in men without clinical leukocytospermia.

Diverse and related 16S rRNA-encoding DNA sequences in prostate tissues of men with chronic prostatitis.

Treatment of chronic prostatitis/chronic pelvic pain syndrome is often empirical because clinical culture methods fail to detect prostate-associated pathogens in >90% of patients. Previously, we tested a variety of specific-microorganism PCRs and began a DNA sequence study after we found that 77% of prostatitis patients were PCR positive for prokaryotic rRNA-encoding DNA sequences (rDNAs) despite negative cultures using optimal techniques. In the present study, 36 rDNA clones from 23 rDNA-positive patients were sequenced. This study represents more than twice the total rDNA sequence and more than twice the number of patients in the previous study. The increased number of patients and clones sequenced allowed enhanced phylogenetic analyses and refinements in our view of rDNA species inhabiting the prostate. A continuum of related rDNAs that might be arbitrarily described as two major groups of rDNAs and several minor groups was found. Sequences termed Pros A, identified in 8 (35%) of 23 rDNA-positive patients, grouped with Aeromonas spp. in phylogenetic studies. Sequences termed Pros B, identified in 17 (74%) of 23 rDNA-positive patients, were distinct from previously reported sequences, although all were >90% similar to known gram-negative bacteria. Of the nine patients for whom multiple rDNAs were sequenced, six had biopsy specimens containing rDNAs from more than one species. Four (17%) patients had rDNAs different from those of the Pros A and Pros B groups. Of these four, one patient had rDNA similar to that of Flavobacterium spp., another had rDNA similar to that of Pseudomonas testosteroni, and two patients had rDNAs <70% similar to known rDNAs. These findings suggest that the prostate can harbor bacteria undetectable by traditional approaches. Most of these diverse sequences are not reported in environments outside the prostate. The sequence similarities suggest adaptation of limited groups of bacteria to the microenvironment of the prostate. Further studies may elucidate the relationship of prostate-associated bacteria to chronic prostatitis/chronic pelvic pain syndrome.

Triangulation end-to-side vasoepididymostomy.

PURPOSE: A technique and the preliminary results of triangulation vasoepididymostomy are described. MATERIALS AND METHODS: Triangulation vasoepididymostomy is performed by placing 3 double-armed 10-zero nylon sutures into the epididymis so that each suture forms 1 side of a triangle. An opening in the tubule is made between the sutures and they are brought inside-out, invaginating the epididymal tubule into the vas deferens. RESULTS: Triangulation end-to-side vasoepididymostomy was performed bilaterally in 12 men, and sperm was found in the postoperative ejaculate of 11 (92%). Operative time was 156 +/- 14 minutes. CONCLUSIONS: Triangulation vasoepididymostomy is reasonably successful in restoring sperm to the ejaculate and should be considered as an alternative method of vasoepididymostomy.

Chronic exposure of cancer-prone mice to low-level 2450 MHz radiofrequency radiation.

The purpose of this study was to determine whether chronic, low-level exposure of mammary-tumor-prone mice to 2450 MHz radiofrequency radiation (RFR) promotes an earlier onset (decreased latency), a greater total incidence, or a faster growth rate of mammary tumors. One hundred C3H/ HeJ mice were exposed in circularly polarized waveguides (CWG) for 18 months (20 h/day, 7 days/wk) to continuous-wave, 2450 MHz RFR at a whole body average specific absorption rate (SAR) of 0.3 W/kg; 100 mice were sham exposed. Before exposure, SARs were determined calorimetrically; during experimentation, SARs were monitored by differential power measurement. All animals were visually inspected twice daily and were removed from the CWG cages for a weekly inspection, palpation, and weighing. From the time of detection, tumor size was measured weekly. Animals that died spontaneously, became moribund, or were killed after 18 months of exposure were completely necropsied; tissues were fixed and subjected to histopathological evaluations. Results showed no significant difference in weight profiles between sham-irradiated and irradiated mice. Concerning mammary carcinomas, there was no significant difference between groups with respect to palpated tumor incidence (sham = 52%; irradiated = 44%), latency to tumor onset (sham = 62.3 +/- 1.2 wk; irradiated = 64.0 +/- 1.6 wk), and rate of tumor growth. In general, histopathological examination revealed no significant differences in numbers of malignant, metastatic, or benign neoplasms between the two groups; a significantly greater incidence of alveolar-bronchiolar adenoma in the sham-irradiated mice was the only exception. In addition, survival analysis showed no significant difference in cumulative percent survival between sham and irradiated animals. Thus, results indicate that under the conditions of this study, long-term, low-level exposure of mammary-tumor-prone mice to 2450 MHz RFR did not affect mammary tumor incidence, latency to tumor onset, tumor growth rate, or animal longevity when compared with sham-irradiated controls.

Bladder petechiae after cystoscopy and hydrodistension in men diagnosed with prostate pain.

PURPOSE: We determined if men with prostate pain syndromes have petechiae in the bladder after hydrodistension. MATERIALS AND METHODS: A total of 60 men with the diagnosis of prostate pain and without bacteriuria underwent cystoscopy and hydrodistension under a general or regional anesthetic. RESULTS: Of the 60 men 35 (58%) had moderate to severe petechiae similar in appearance to women with interstitial cystitis after hydrodistension. Men with moderate to severe bladder petechiae had fewer leukocytes in expressed prostatic secretions, smaller bladder capacities and less often testicular pain than men with more normal appearing bladders after hydrodistension. Symptomatic improvement 2 to 6 weeks after hydrodistension was more common in men with moderate to severe petechiae than in those with fewer petechiae. Absence of rectal pain predicted symptomatic improvement after hydrodistension. CONCLUSIONS: We suggest that bladder petechiae, and possibly interstitial cystitis or a related condition, may be more frequently associated with prostate pain syndromes in men than previously appreciated.

Distribution and mobility of the tetracycline resistance determinant tetQ.

We tested 34 American Type Culture Collection (ATCC) and 168 clinical bacterial isolates, from the human urogenital and oral tracts and streptococci isolated from cows with mastitis, for the presence of the tetQ gene using a polymerase chain reaction (PCR) assay and DNA-DNA hybridization. The identities of PCR products were confirmed by Southern blot hybridization of whole-cell DNA. Eleven of the ATCC strains were positive for tetQ, including five Bacteroides spp., five Prevotella spp. and a single isolate of Mitsuokella multiacidus. Twenty-eight (29%) of the 95 clinical Gram-negative isolates carried the tetQ gene, while eight (11%) of the 73 clinical Gram-positive isolates carried the tetQ gene. This is the first description of tetQ in Gram-positive species. All isolates except one Peptostreptococcus sp. carried tetQ integrated into the chromosome. The tetQ gene could be transferred from Prevotella bivia, Bacteroides ovatus, Bacteroides fragilis, Bacteroides vulgatus and Bacteroides distasonis into an Enterococcus faecalis recipient at frequencies of 10(-7)-10(-9) per recipient. In contrast, tetQ failed to transfer from two isolates of Prevotella intermedia, two isolates of Porphyromonas gingivalis, one isolate of Mobiluncus curtisii and one isolate of Peptostreptococcus sp. The latter two are Gram-positive species. The PCR assay was used to screen 198 proteinase K-treated biopsies of prostate, periprostate and bladder from 84 men with prostatitis. Thirty-four (40%) of the patients had one or more positive samples, suggesting that the PCR assay could be of value in screening patient material directly for the presence of bacteria.

The effect of pharmacologic erection on the dorsal nerve of the penis.

The dorsal nerve of the penis (DNP) is the primary source of afferent somatic input from the penis and is critical in the male sexual functions of erection and ejaculation. Using genitourinary electrodiagnostic techniques, this study was conducted to investigate the effect of pharmacologic erection on the DNP. Three tests were administered, and the changes in the DNP between flaccid, stretched, and erect states were examined. Calculated nerve conduction velocity (cNCV) measurements of the DNP increased with pharmacologic erection because mechanical straightening of the nerve allowed for a more precise measurement of nerve length. The latencies of the cortical evoked response and the bulbocavernosus reflex were not significantly changed with stretch or pharmacologic erection. In the evaluation of impotence, cNCV DNP measurements should be performed on the erect penis.

Randomized clinical trial of an absorbable stent for vasectomy reversal.

PURPOSE: More successful methods of vasectomy reversal would benefit those undergoing this treatment and might also increase the popularity of vasectomy. We conducted a randomized, prospective clinical trial of vasectomy reversal methods, comparing a new absorbable stent with 2-layer reattachment. MATERIALS AND METHODS: We studied 116 men seeking vasectomy reversal between November 1990 and March 1994. Data were analyzed primarily by intention to treat. RESULTS: Patients in the stent (64 cases) and no stent (52) randomized groups were similar in age distribution, age of spouses, years since vasectomy, proportion who had undergone prior vasectomy reversal and proportion who had previously achieved pregnancy. Operation time was more than 19 minutes shorter in the stent randomized group (p = 0.006). Fewer patients in the stent than the no stent group had patent vasovasostomies (81.0 versus 89.6%, respectively, p = 0.2) postoperatively. Fewer stent randomized patients had motile sperm (76.2 versus 81.3%, respectively, p = 0.5) and normal total motile sperm counts (49.2 versus 52.1%, respectively, p = 0.8) than did those without a stent. Conception occurred in 22 and 51% of all couples in the stent and no stent groups. The relative risk of conception among those in the stent group was 0.42 (95% confidence interval 0.24 to 0.71, p = 0.002). A Mantel-Cox log-rank test comparing pregnancies in each group according to the number of postoperative months revealed that the no stent group achieved more earlier pregnancies (p = 0.003). CONCLUSIONS: The 2-layer microscopic vasovasostomy results in greater pregnancy rates than vasovasostomy using the absorbable stent.

Post-traumatic priapism treated with selective cavernosal artery ligation.

Two cases of post-traumatic high-flow priapism after blunt trauma to the penis are presented. Diagnosis of high-flow priapism was accomplished with the use of both color-flow Doppler ultrasound and arteriography, which demonstrated arteriocorporal fistulas. In each patient, angiographic embolization was attempted but abandoned because the distal artery feeding the fistula could not be safely catheterized. Both patients were definitively treated with surgical ligation of the arteriovenous fistula, guided by intraoperative ultrasound. Operative management with arterial ligation provides a safe, selective, and effective alternative treatment to embolization. Two surgical approaches are discussed, one extracorporal and the other transcorporal. We believe that although transcorporal dissection poses increased risks, it is appropriate for arterial priapism of prolonged duration, especially if a well-formed vascular pseudocapsule is identified.