Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men

OBJECTIVE: Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. MATERIALS AND METHODS: Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v) with the cryoprotector containing glycerol (Test yolk buffer). One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI), while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III) (group II - GII). The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog%) and percentage of live spermatozoa (Vit%). After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h). The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. RESULTS: There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p < 0.001). Values of Prog% and Vit% were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog% among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006). After cryoprotector removal, Prog% has been kept unaltered, in both groups, during the first 3 hours of incubation, although being superior in group I (p = 0,04). There was a significant decrease in Prog% after 24 hours of incubation, in both groups (p < 0,01). CONCLUSION: For human semen samples with low initial quality, freezing through vapor technique or through the automated technique showed to be equivalent in regarding recovery of live spermatozoa with progressive motility. The effects of dilution and centrifugation to remove the cryoprotector had a negative impact only in samples frozen through the automated technique. In both techniques, progressive motility is kept constant during the first 3 hours after thawing and removal of the cryoprotector, but is drastically diminished by the end of an incubation period of 24 hours.Universidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Division of UrologyUNIFESP, EPM, Division of UrologySciEL

Prognostic relevance of the histological subtype of renal cell carcinoma

OBJECTIVE: According to several studies, when the histological subtype of renal cell carcinoma is established it is possible to attribute a different life expectancy to each patient. We analyzed the prognostic significance of the histological subtype in renal cell carcinoma. MATERIALS AND METHODS: The authors retrospectively analyzed the follow-up of 230 patients after radical or conservative renal surgery. The histological characteristics of the different subtypes of tumor were obtained and the disease-free and cancer-specific survival curves for the clear cell, cromophobic, papillary, collecting duct (Bellini) subtypes and those with sarcomatoid differentiation were individualized. RESULTS: The disease-free and cancer-specific survival rates for clear cell tumors were 76.6% and 68.0% respectively, 71.2% and 82.1% respectively for the cromophobic type, 71.1% and 79.8% respectively for the papillary type, 26.9% and 39.3% respectively for the sarcomatoid type, and 0.0% and 0.0% respectively for the collecting ducts (p < 0.001). CONCLUSION: The histological subtypes of renal tumors can stratify patients into different prognostic groups only when the sarcomatoid differentiation is present

Pudendal nerve latency time in normal women via intravaginal stimulation

INTRODUCTION & OBJECTIVES: Studies of motor conduction for the efferent functional assessment of the pudendal nerve in women with pelvic dysfunctions have been conducted through researching distal motor latency times. The transrectal approach has been the classic approach for this electrophysiological examination. The objective of the present study is to verify the viability of the transvaginal approach in performing the exam, to establish normal values for this method and to analyze the influence of age, stature and parity in the latency value of normal women. MATERIALS AND METHODS: A total of 23 volunteers without genitourinary pathologies participated in this study. In each, pudendal motor latency was investigated through the transvaginal approach, which was chosen due to patient s higher tolerance levels. RESULTS: The motor response represented by registering the M-wave was obtained in all volunteers on the right side (100%) and in 13 volunteers on the left side (56.5%). The mean motor latency obtained in the right and left was respectively: 1.99 ± 0.41 and 1.92 ± 0.48 milliseconds (ms). There was no difference between the sides (p = 0.66). Latency did not correlate with age, stature or obstetric history. The results obtained in the present study were in agreement with those found by other researchers using the transrectal approach. CONCLUSION: The vaginal approach represents an alternative for pudendal nerve distal motor latency time, with similar results to those achieved through the transrectal approach. Normative values obtained herein might serve as a comparative basis for subsequent physiopathological studies.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Departments of Urology and NeurologyUNIFESP, EPM, Departments of Urology and NeurologySciEL

Magnetic resonance imaging urodynamics: technique development and preliminary results

OBJECTIVES: In this preliminary study we report the development of the video urodynamic technique using magnetic resonance imaging (MRI). MATERIALS AND METHODS: We studied 6 women with genuine stress urinary incontinence, diagnosed by history and physical examination. Urodynamic examination was performed on multichannel equipment with the patient in the supine position. Coughing and Valsalva maneuvers were performed at volumes of 150, 250 and 350 mL. Simultaneously, MRI was carried out by using 1.5 T GE Signa CV/i high-speed scanner with real time fluoroscopic imaging possibilities. Fluoroscopic imaging was accomplished in the corresponding planes with T2-weighted single shot fast spin echo sequences at a speed of about 1 frame per second. Both studies were recorded and synchronized, resulting in a single video urodynamic examination. RESULTS: Dynamic MRI with cine-loop reconstruction of 1 image per second demonstrated the movement of all compartment of the relaxed pelvis during straining with the concomitant registration of abdominal and intravesical pressures. In 5 patients, urinary leakage was demonstrated during straining and the Valsalva leak point pressure (VLPP) was determined as the vesical pressure at leak subtracted from baseline bladder pressure. Mean VLPP was 72.6 cm H2O (ranging from 43 to 122 cm H2O). CONCLUSIONS: The concept of MRI video urodynamics is feasible. In a clinical perspective, practical aspects represent a barrier to daily use and it should be recommended for research purposes.Federal University of São Paulo Department of UrologyUNIFESP, Department of UrologySciEL

Effects of curcumin in an orthotopic murine bladder tumor model

Cigarette smoking (CS) is the main risk factor for bladder cancer development. There are more than 100 carcinogens present in cigarette smoke. Among the potential mediators of CS-induced alterations is nuclear factor-kappa (NF-&#954;B), which is responsible for the transcription of genes related to cell transformation, tumor promotion, angiogenesis, invasion and metastasis. Curcumin is a polyphenol compound derived from Curcuma longa that suppress cellular transformation, proliferation, invasion, angiogenesis, and metastasis by down regulating NF-&#954;B and its regulated genes. The aim of our study was to assess the effects of curcumin in bladder urothelial carcinoma. We studied the effects of curcumin in vitro and in vivo using the orthotropic syngeneic bladder tumor animal model MB49. Curcumin promotes apoptosis of bladder tumor cells in vitro. In vivo tumors of animals treated with curcumin were significantly smaller as compared to controls. Using immunohistochemistry, we demonstrated a decrease in the expression of Cox-2 by 8% and Cyclin D1 by 13% in the animals treated with curcumin; both genes regulated by NF-&#954;B and related to cell proliferation. In this study, we showed that curcumin acts in bladder urothelial cancer, possibly dowregulating NF-&#954;B-related genes, and could be an option in the treatment of urothelial neoplasms. The results of our study suggest that further research is warranted to confirm our findings

Early experience with targeted therapy and dendritic cell vaccine in metastatic renal cell carcinoma after nephrectomy

PURPOSE: Metastatic renal cell carcinoma (RCC) is one of the most treatment-resistant malignancies and nephrectomy, isolated or combined with systemic chemotherapy typically has limited or no effectiveness. We report our initial results in patients treated with the association of molecular targeted therapy, nephrectomy, and hybrid dendritic-tumor cell (DC) vaccine. MATERIALS AND METHODS: Two male patients diagnosed with metastatic RCC were selected for the study. They were treated with the triple strategy, in which sunitinib (50 mg per day) was given for 4 weeks, followed by radical nephrectomy after two weeks. DC vaccine was initiated immediately after surgery and repeated monthly. Sunitinib was restarted daily after 2 to 3 weeks of surgery with a 7-day interval every 4 weeks. RESULTS: Both patients had complete adherence to the proposed treatment with DC vaccine therapy combined with sunitinib. Follow-up in these patients at 9 and 10 months demonstrated a stable disease in both, as shown by imaging and clinical findings, with no further treatment required. CONCLUSION: The immune response obtained with DC vaccine combined with the antiangiogenic effect of sunitinib and the potential benefits of cytoreductive nephrectomy in advanced disease could represent a new option in the treatment of metastatic RCC. Further prospective trials are needed not only to elucidate the ideal dosing and schedule, but also to better define the proof-of-concept proposed in this report and its role in clinical practice

The use of immunohistochemistry for diagnosis of prostate cancer

PURPOSE: Atypical glands (ASAP) are diagnosed in 5.0% of prostate biopsies, and cancer identification in a rebiopsy is higher than 40.0%. The use of antibodies to mark basal cells is currently a common practice, in order to avoid rebiopsies. There has been no reported study that has reviewed characteristics of radical prostatectomies (RPs) when immunohistochemistry (IHC) was necessary for definitive diagnosis. MATERIALS AND METHODS: Out of 4127 biopsies examined from 2004 to 2008, 144 (3.5%) were diagnosed with ASAP. IHC was performed using antibody anti-34ßE12 and p63. The results of surgical specimens of 27 patients treated by RP after the diagnosis of prostate cancer (PC) was made using IHC (Group 1) were compared with 1040 patients where IHC was not necessary (Group 2). RESULTS: IHC helped to diagnose PC in 103 patients (71.5%). Twenty-seven (26.2%) underwent RP. In Group 1, two (7.4%) adenocarcinomas were insignificant versus 29 (2.9%) for Group 2. Patients from Group 1 were younger (p = 0.039), had lower Gleason scores (GS) (p < 0.001), lower percentage of Gleason pattern 4 (p < 0.001), and smaller tumors (p < 0.001). CONCLUSION: The use of IHC did not lead to diagnosis of insignificant tumors as illustrated by absence of differences in pathological stage or positive surgical margins in men submitted to RP. Therefore, our results suggest that this modality should be routinely used for a borderline biopsy and ASAP cases

Study of the efficacy of Korean Red Ginseng in the treatment of erectile dysfunction

Aim: To examine the treatment efficacy of Korean Red Ginseng (KRG) in impotent men with erectile dysfunction (ED). Methods: A total of 60 patients presenting mild or mild to moderate ED were enrolled in a double-blind, placebo-controlled study in which the efficacies of KRG and a placebo were compared. the patients received either 1 000 mg (3 times daily) of KRG or a placebo. Results: the five-item version of the International Index of Erectile Function (IIEF-5) score after the treatment was significantly higher in the KRG group compared with that before the treatment (from 16.4 +/- 2.9 to 21.0 +/- 6.3, P 0.05). in the KRG group, 20 patients (66.6%), reported improved erection, significant in the global efficacy question (P 0.05). Conclusion: Our data show that KRG can be an effective alternative to the invasive approaches for treating male ED.Univ São Paulo, Div Urol Clin, Sector Sexual Med, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Urol, São Paulo, BrazilUniversidade Federal de São Paulo, Discipline Urol, São Paulo, BrazilWeb of Scienc

Search and identification of spermatozoa and spermatids in the ejaculate of non-obstructive azoospermic patients

OBJECTIVE: To search and to identify spermatozoa and spermatids, present in the ejaculate of non-obstructive azoospermic patients. MATERIALS AND METHODS: 27 patients, aged between 18 and 48 years, with initial diagnosis compatible with non-obstructive azoospermia, underwent up to 3 seminal samples, with assessment of macroscopic and microscopic parameters differentiated for each sample. In the first sample, 5 µL of semen were analyzed in a Horwell chamber in order to assess the presence or absence of spermatozoa. The procedure was repeated with 2 other aliquots. In the absence of spermatozoa, the entire sample was transferred to a conic tube and following centrifugation the sediment was freshly analyzed. The second seminal sample was collected only when no spermatozoa were found in the first sample and the research was performed in the same way. In cases where spermatozoa were not seen, the sample was centrifuged and the obtained sediment was stained by the panoptic method and observed under common light microscopy (1250X). The third seminal sample was collected only in cases when patients had not shown spermatozoa in the first and second seminal samples. RESULTS: 4/27 (14.8%) patients presented spermatozoa in the first seminal sample and 6/23 (26.1%), in the second seminal sample. No spermatozoa were seen in the third sample, however, 11/17 (64.7%) presented spermatids. CONCLUSION: In clinical situations where the initial diagnosis is non-obstructive azoospermia, one single routine seminal analysis is not enough to confirm this diagnosis and the analysis of the centrifuged sediment can have relevant clinical consequences. Among patients considered non-obstructive azoospermic, when duly assessed, 37% presented spermatozoa and 64.7%, spermatids.Universidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Division of UrologyUNIFESP, EPM, Division of UrologySciEL

Abnormal expression of MDM2 in prostate carcinoma

Mutation of p53 is rare in localized prostate carcinoma the oncoprotein MDM2, whose gene has a response element for p53, promotes the degradation of p53 protein and inhibits its transcriptional activation of genes related to cell cycle arrest and apoptosis, constituting a negative feedback control. We studied p53 and MDM2 expression by immunohistochemistry and looked for mutations in p53 exons 5 to 8 by polymerase chain reaction-single strand conformational polymorphism in 118 patients submitted to radical prostatectomy for localized prostate cancer. in 28 cases, we studied cell proliferation by immunohistochemistry, using antibody for Ki-67, and apoptosis by the deoxynucleotidyl transferase mediated dUTP biotin nick end labeling technique. Although no p53 mutations were found, p53 protein was detected in 31.4% of the cases, and these cases had higher Gleason scores (P = .03) and more advanced tumor stages (P = .02). MDM2 was overexpressed in 40.7% of the cases, and these: cases had greater tumor volumes (P = .001). Tumors that were positive for both p53 and MDM2 were larger (P = .003) and of more advanced stage (P = .03). Within the 28-case subset, the proliferative index was higher among MDM2-positive tumors (P = .046), and the apoptotic index was lower among p53-positive tumors (P = .01). We conclude that, although p53 mutation is a rare event in prostate carcinogenesis, the detection of p53 protein by immunohistochemistry is common and is associated with decreased apoptosis and increased histologic grade and tumor stage. We also conclude that the overexpression of MDM2 has a role in prostate carcinogenesis, being frequently detected and associated with increased cell proliferation and tumor volume. Finally, we propose that the MDM2-positive/53-positive phenotype identifies prostate cancers with aggressive behavior.Hosp Sirio Libanes, Lab Surg & Mol Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Surg, Div Urol, São Paulo, BrazilUniv São Paulo, Dept Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pathol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Surg, Div Urol, São Paulo, BrazilWeb of Scienc