Comparative analysis of the germ cell markers c-KIT, SSEA-1 and VASA in testicular biopsies from secretory and obstructive azoospermias.

Testicular biopsy is needed to confirm diagnosis in azoospermic patients and to recover spermatozoa, if possible. This report aims to quantitatively analyse the germline markers stage-specific embryonic antigen (SSEA-1), c-KIT and VASA in testicular biopsies with distinct azoospermic aetiologies. Twenty-three testicular biopsies were analysed by flow cytometry and RT-qPCR for c-KIT, SSEA-1 and VASA. In all the Sertoli cell-only (SCO) samples, significantly lower VASA mRNA expression and fewer VASA+ cells were found compared with obstructive controls. Maturation arrest (MA) cases showed significant differences only with the non-mosaic SCO samples when compared for VASA mRNA expression and percentage of VASA+ cells, but not with the mosaics. However, the normalized VASA-KIT parameter obtained by subtracting the percentage of c-KIT+ cells from the percentage of VASA+ cells showed significant differences between the MA and all the SCO samples. RT-qPCR consistently found differences for the VASA expression between SCO mosaic and non-mosaic samples. However, by flow cytometry, only VASA-KIT showed significant differences between them. Conversely, the percentage of SSEA-1+ cells revealed no inter-group differences. In conclusion, testicular biopsies display different expression profiles for c-KIT and VASA depending on the azoospermic aetiology. These results can be used as a complementary tool to create new molecular categories for diagnoses in azoospermic patients, particularly useful to discriminate between mosaic and non-mosaic SCO patients.

[Value of transrectal FNA in the diagnosis of prostate cancer in elderly patients]. / Valor de la PAAF transrectal en el diagnóstico del cáncer de próstata en pacientes de edad avanzada.

OBJECTIVES: To identify if there is a group elderly patients with clinical suspicion of prostate cancer in which pathological confirmation may be unnecessary, and if prostatic transrectal fine needle aspiration (FNA) may be a useful diagnostic tool for old patients. MATERIAL AND METHODS: A total of 72 patients aged 75-93 years were evaluated by means of prostatic transrectal FNA. Antibiotic prophylaxis, analgesia or cessation of anticoagulant therapy were not necessary. RESULTS: In 35 patients (48.6%) cytological diagnosis was positive for prostatic adenocarcinoma, whereas in 37 cytology was negative for cancer. In 100% of patients with PSA > 30 ng/ml or with PSA > 20 ng/ml and suspicious digital rectal examination FNA results were positive for cancer. 4.1% minor and 1.3% major (acute prostatitis) complications after FNA were observed. CONCLUSIONS: In male patients older than 75 years with PSA > 30 ng/ml or with PSA > 20 ng/ml and suspicious digital rectal examination, histological confirmation of carcinoma by prostatic biopsy may be not necessary, because of the high probability of a positive result. When histological confirmation of prostatic carcinoma is required in elderly patients, transrectal prostatic FNA is a valid alternative to transrectal biopsy, due to its excellent tolerance and low complication rate.

[Changes in sexual hormones in a male population over 50 years of age. Frequency of low testosterone levels and risk factors]. / Cambios en las hormonas sexuales en varones mayores de 50 años. Prevalencia de niveles bajos de testosterona y factores de riesgo.

OBJECTIVES: To investigate the changes in sexual hormones in a selected male population older than 50 years of age. To assess the frequency of biochemical hypogonadism and which factors are related to testosterone levels. PATIENTS AND METHODS: A Cross-sectional study was carried out on 230 Spanish men older than 50 years of age. Blood tests were performed including: total testosterone, SHBG, calculated free testosterone, dehidroepiandrosterone sulfate, androstendione, estradiol, bioavailable estradiol, FSH, LH, and prolactin. Clinical and socio-demographic backgrounds were investigated. The frequency of biochemical hypogonadism was established using total and free testosterone levels as diagnostic criteria. Factors that may influence testosterone levels were evaluated by univariate and multivariate statistical analysis, and a logistic regression model was used to determine which factors can predict biochemical hypogonadism according to free testosterone levels. RESULTS: Age was associated with a significant decrease (p < 0.05) in total testosterone (0.6% per year), free testosterone (1.3% per year), dehydroepiandrosterone sulfate (1.8% per year) and bioavailable estradiol (0.69% per year). Moreover, an increase in SHBG, LH, and FSH was observed (p < 0.05). According to total testosterone levels, 4.8% of the men were hypogonadal, whereas 24.8% were hypogonadal when free testosterone was considered. In the univariate analysis, obesity, diabetes mellitus and hyperlipemia were related to lower total testosterone levels, while free testosterone levels were lower in men with sedentary life, lower levels of education, obesity or diabetes mellitus. In the multivariate analysis age, diabetes mellitus and obesity were inversely related to total and free testosterone levels. Free testosterone was also inversely related to hyperlipemia. For biochemical hypogonadism, simple logistic regression analysis selected age, sedentary life, obesity and diabetes mellitus. In the multivariate analysis age, obesity and diabetes mellitus had significant independent prognostic value. CONCLUSIONS: Starting from 50 years of age, a significant age-related decrease in total testosterone, free testosterone, dehydroepiandrosterone sulfate and bioavailable estradiol is observed. The frequency of biochemical hypogonadism is higher when free testosterone levels are used for diagnosis. Total testosterone levels were related to age, diabetes mellitus and obesity. Free testosterone was related to age. diabetes mellitus, obesity and hyperlipemia. The probability of suffering low free testosterone levels increases with age, diabetes mellitus and/or obesity.

[Bilateral testicular cancer: a report of four cases]. / Cancer testicular bilateral: presentación de cuatro casos.

Bilateral testicular cancer represents from 2 to 5% of all testicle tumors, even though this prevalence nowadays is increasing. The median age of presentation was from 15 to years and in approximately 75% of them occurred metachronously. There used to be concordance in histological findings between both testicles, being seminoma the most common histological type. The most important risk factor in the development of testicular cancer is the presence of intratubular germ cell neoplasia. Radical orquiectomy is the treatment to choose, although in some carefully selected patients testis-sparing surgery may be considered. We present four cases attended in our medical service, we analize the risk factors, management of tumors and a medical literature review is done.

[Evaluation of an immunoassay kit to measure free testosterone]. / Evaluación de una técnica de inmunoanálisis para la determinación de testosterona libre.

INTRODUCTION AND OBJECTIVES: The best indicators to the diagnosis of hypogonadism are free and bioavailable testosterone circulating levels. Free and bioavailable testosterone measurements are complex. However, simple kits for direct measurement of free testosterone by analog immunoassay are available. We examined the utility of an enzymoimmunoassay kit for free testosterone measurement. MATERIAL AND METHOD: One hundred thirty-three healthy males were included. Total testosterone, SHBG, albumin and free testosterone was measured. We used two different methods to free testosterone estimation: direct measurement by enzymoimmunoassay and mathematical calculation with Vermeulen's formula, which uses albumin concentration, total testosterone and SHBG to calculate free testosterone (method recommended by the International Society for the Study of the Aging Male). We compared the two methods means values and a linear regression study was performed. RESULTS: Mean age was 37 +/- 11 years. Mean serum concentration for total testosterone was 21.43 +/- 6.8 nm ol/L. The mean value for free testosterone measured by direct and mathematical method was 0.0508 +/- 0.0118 nmol/L and 0.474 +/- 0.123 nmol/L respectively. In linear regression study exists a positive correlation between both methods (p< 0.05), although correlation coefficient is very low (r = 0.25). CONCLUSIONS: There are significant statistical differences between the measurements of free testosterone by direct and mathematical methods. Although certain correlation is observed, this is very low. In conclusion, free testosterone measurement by enzymoimmunoassay is not reliable.

Intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

To assess if testicular sperm cryopreservation is a valid alternative to repetition of testicular sperm retrieval techniques, results of a cryopreservation technique in pills have been retrospectively analyzed. Enough motile spermatozoa for ICSI were obtained in 172 from 190 (90.5%) frozen-thawed testicular sperm samples. Overall, 249 couples underwent 390 ICSI cycles, 156 using fresh and 234 using cryopreserved testicular sperm. Mean two-pronuclear fertilization rates per cycle were not significantly different after ICSI with fresh (62.0%) or with cryopreserved (63.2%) spermatozoa. Mean embryo cleavage rate per cycle was higher in the fresh (90.6%) than in the cryopreserved (84.6%) group (P = 0.016). However, clinical pregnancy rates per cycle (28.2% with fresh vs 27.8% with cryopreserved), implantation rates (12.2% vs 13.1%) and ongoing pregnancy rates per cycle (22.4% vs 21.8%) were not significantly different. Cryopreservation of testicular spermatozoa is an effective technique that can be used both in obstructive and in non-obstructive azoospermia.

Pregnancy in an azoospermic patient with markedly elevated serum follicle-stimulating hormone levels.

OBJECTIVE: To assess the possibility of achieving a pregnancy in an azoospermic patient with markedly elevated serum FSH level. DESIGN: A case report. SETTING: In vitro fertilization program at the Instituto Valenciano de Infertilidad. PATIENT: An azoospermic patient with small testes and serum FSH level (38.7 mIU/mL) higher than three times normal. Testicular biopsy revealed Sertoli cell-only syndrome with focal spermatogenesis. INTERVENTIONS: Intracytoplasmic microinjection of testicular tissue-extracted spermatozoa. MAIN OUTCOME MEASUREMENTS: Fertilization rate, cleavage rate, clinical pregnancy. RESULTS: Eight of 11 (73%) intact oocytes showed two pronuclei. All of them cleaved normally. Four embryos were replaced into the uterine cavity and the other four were cryopreserved. A twin clinical pregnancy was achieved. CONCLUSION: Spermatozoa may be present in testicular biopsy specimens of azoospermic patients with severe spermatogenic failure despite markedly elevated serum FSH level. These patients can be fertile after intracytoplasmic testicular sperm microinjection.

Fertilization after intracytoplasmic sperm injection with cryopreserved testicular spermatozoa.

OBJECTIVE: To assess the possibility of cryopreserving testicular tissue extracted sperm for intracytoplasmic sperm injection (ICSI). DESIGN: A report of two cases. Our study was approved by the Ethical Committee at the Instituto Valenciano de Infertilidad. SETTING: In vitro fertilization program at the Instituto Valenciano de Infertilidad. PATIENTS: Two azoospermic patients with severe spermatogenic failure but with focal spermatogenesis on testicular biopsies. In both cases, a first ICSI attempt with fresh testicular biopsy extracted sperm was unsuccessful. INTERVENTIONS: Cryopreservation of testicular spermatozoa in 100-micro L "pills." Intracytoplasmic sperm injection with thawed testicular spermatozoa. MAIN OUTCOME MEASUREMENTS: Fertilization rate, cleavage rate, embryo quality, clinical pregnancy. RESULTS: Fertilization rates were 36 percent and 100 percent after ICSI with fresh testicular spermatozoa, and 63 percent and 57 percent after ICSI with cryopreserved testicular sperm. In both cases, cleavage rates and embryo quality were similar when using fresh and cryopreserved testicular spermatozoa. No clinical pregnancies were achieved. CONCLUSION: High fertilization rates can be obtained after ICSI with frozen-thawed testicular tissue extracted spermatozoa. Cryopreservation of testicular sperm may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testicle.

Screening for AZF deletion in a large series of severely impaired spermatogenesis patients.

Recent investigations have pointed to a high prevalence of Y chromosome submicroscopic deletions in men with severely impaired spermatogenesis. We report on the incidence in 128 infertile men, in whom karyotype, sperm count, and hormonal parameters were evaluated. Patients with abnormal karyotype (other than an abnormal Y chromosome) or sperm concentration of more than 2 million/mL were excluded. Genomic DNA was extracted from the peripheral leukocytes of 57 men with azoospermia and 71 with severe oligospermia. Molecular analysis was performed by 3 multiplex polymerase chain reactions using a set of 9 sequence tagged sites (STSs) from 3 different regions of the Y chromosome: AZFa, AZFb, and AZFc. In 7% of the studied patients Yq microdeletions were detected, with a high prevalence in men with azoospermia (14%). No deletions were detected in the AZFa region. Deletions were present in AZFb, AZFc, or both regions. The deletion observed in 1 patient that did not overlap with the DAZ region demonstrates that genes other than DAZ may also be involved in the pathogenesis of some subsets of male infertility. Furthermore, common Yq deletions present different testicular pictures, suggesting that some unknown factors may be disturbing spermatogenesis. Because men with severe infertility suffer a high risk of Y chromosome deletion, screening for these men is recommended prior to treatment with assisted reproduction.

[Value of isotopic cystography in the diagnosis of vesicoureteral reflux in childhood]. / Valor de la cistografía isotópica en el diagnóstico del reflujo vesicoureteral en la infancia.

We have studied 62 children with urinary infection simultaneously by means of micturitional cystoureterography (MCU) and indirect isotopic cystography. We confirm a correlation between the results of the MCU and in the indirect isotopic cystography in 86% of the ureters explored, whilst the results of the MCU and of the direct isotopic cystography coincided in 92% of the ureters explored. We discuss the advantages and drawbacks of isotopic cystography, concluding that it is a suitable method for the diagnosis and evolutive control of vesicoureteral reflux in infancy.

[Value of prostate-specific antigen density and transitional-zone prostate-specific antigen density in the diagnosis of prostate cancer]. / Valor de la densidad del antígeno prostático específico y de la densidad del antígeno prostático específico de la zona transicional en el diagnóstico del cáncer de próstata.

INTRODUCTION: With serum prostate specific antigen (PSA) levels of 4.1 to 10 ng/ml a significant number of patients are unnecessary subjected to biopsy. We try to determine if the calculation of prostate specific antigen density (PSAD) and prostate specific antigen density of the transition zone (PSADTZ) improve the capacity to discriminate between non-malignant disease and prostate cancer. METHODS: A prospective study including 314 males with PSA levels between 4.1 and 10 ng/ml is reported. Transrectal ultrasonography and prostatic biopsy were performed in all of them and total prostate and transition-zone volumes were calculated. PSA density and PSA density of the transition zone were calculated for each patient. Receiver operating characteristics (ROC) curves for PSA, PSAD and PSADTZ were constructed for all the patients and for those patients with digital rectal examination unsuspicous of malignancy, determining the sensitivity and specificity for several cutoff values. RESULTS: The area under the curve for both, PSAD and PSADTZ, were greater than for PSA (p < 0.05), without any significant differences between PSADTZ and PSAD. The cutoff value of greatest diagnostic efficiency for PSAD was 0.17 ng/ml/cc (71.4% sensitivity and 55.7% specificity), while it was 0.41 ng/ml/cc for DPSATZ (70% sensitivity and 61.5% specificity). For those cases of normal digital rectal examination, no differences were observed between PSA and PSAD but they were between PSA and PSADTZ. In any event, the area under ROC curves was always less than 0.7, and, in order to avoid a large number of biopsies (high specificity), a large number of cancers are left without diagnosis (low sensitivity). CONCLUSIONS: We conclude that PSAD and PSADTZ are not excessively useful for adequately discriminating between patients with prostate cancer and those with non-malignant disease, particularly when digital rectal examination is normal.

[The Ask-Upmark kidney: a form of reflux nephropathy]. / El riñón de ASK-Upmark: una forma de nefropatía del reflujo.

We present six female patients aged 14 to 47, all diagnosed histopathologically as suffering from Ask-Upmark Kidney and who had clinical manifestations of severe arterial tension associated with urinary infection in four cases. Mictional Cystourethrography was carried out in all cases, and four of them displayed vesicoureteral reflux in the small kidney. Although this pathology has classically been considered a congenital malformation in the context of renal hypoplasias (segmental hypoplasia), the observation of glomerular traces with PAS staining in the renal segments regarded classically as "aglomerular" and the frequent association of this pathology with vesicoureteral reflux point significantly towards Ask-Upmark Kidney being a form of reflux nephropathy. Nephrectomy cured the arterial hypertension in half the cases, and the factors with prognostic importance in this respect are commented upon.

[Cystic lymphangioma of the adrenal gland in adults. Review of the literature and report of a new case]. / Linfangioma quístico de la glándula suprarrenal del adulto. Revisión de la literatura y aportación de un nuevo caso.

Presentation of one case of cystic lymphangioma of the suprarenal gland in a 34-year-old female patient with a background of lower urinary tract infections, presenting with continuous and aggravating pain in left lumbar area and with no other associated symptomatology, diagnosed with ultrasound, IVU and CAT. A review of the literature is made. The present study pretends to give an overall view of a rare and benign disease, commenting upon the difficulties found in its diagnosis and the different therapeutical options.

[Renal dysplasia and vesico-ureteral reflux]. / Displasia renal y reflujo vesicoureteral.

We studied two groups of patients in order to evaluate the frequency of association between renal dysplasia and vesicoureteral reflux. The first was composed of 19 patients with dysplastic kidneys, 15 of whom were studied by means of micturition cystourethrography. The second group was made up of 86 patients with 124 kidneys with reflux. In 16 of these we were able to obtain a histopathological diagnosis of the renal lesions (12 nephrectomies, 4 biopsies). In three cases the renal dysplasia was associated with a vesicoureteral reflux, which represents an incidence of reflux in 20% of the patients with renal dysplasia, and an incidence of dysplasia in 19% of the kidneys with reflux examined histopathologically. We contrast these results with the literature, discussing the pathogenic hypotheses that enable us to explain this association as well as the clinical consequences thereof.

[Study of oligosaccharides in human urothelial membrane by means of lectins]. / Estudio de los oligosacáridos de la membrana del urotelio humano mediante lectinas.

We have used four lectins (PHA-L, HPA, RCA-120 and PNA) to characterise the oligosaccharides present in the membrane of the different cell layers of the human vesical urothelium, obtained from 27 normal biopsies of vesical mucus. In all the patients studied the PHA-L and HPA lectins showed an affinity for all the urothelial layers, and a progressive increase in fluorescence is observed from the basal to the surface stratum. The pattern observed when using lectin RCA-120 was a positive marking of all the urothelial layers studied, with a homogeneous intensity in all of them. The PNA lectin did not show reactivity to the normal urothelium in none of the cases studied. We comment on the applications of this type of study both for characterizing the glycid composition of the urothelial cell membrane and for ascertaining its variations in accordance with cell differentiation.

[Treatment of advanced prostatic carcinoma with ketoconazole. Our experience in 10 cases]. / Tratamiento del carcinoma prostático avanzado con ketoconazol. Nuestra experiencia en diez casos.

We have treated 10 patients with histologically diagnosed prostate carcinoma, in states C and D, with orally administered dosage of 1200 mg./day of ketoconazol, with the following results: Four patients were excluded through displaying gastrointestinal intolerance in the first days of treatment. Three patients treated had the dosage reduced to half due to poor tolerance, but no improvement was noted. Another three patients tolerated the treatment for four months and displayed transitory remission of ostealgia, which permitted the withdrawal of analgesics. In these patients the treatment was suspended due to digestive intolerance and to the appearance of a clinical picture compatible with suprarrenal corex hypoadrenalism.

[Percentage of tumor in prostatic biopsy cylinders as prognosis factor of organ-limited disease in candidates for radical prostatectomy]. / El porcentaje de tumor en cilindros de biopsia prostática como factor pronóstico de enfermedad órganoconfinada en pacientes candidatos a prostatectomía radical.

UNLABELLED: The present study analyzes the prognostic influence of tumor percentage in cylinders of prostatic biopsy in this patient group. MATERIAL AND METHODS: Retrospective study of 68 patients with a diagnosis of adenocarcinoma, clinical stage T1-T2 who had undergone a radical prostatectomy from May 1997 to october 2000. Following preoperative parameters were analyzed: age, PSA, clinical staging, Gleason and six cylinders of ultrasonography-leaded transrectal prostatic biopsy, studying the amount of positive biopsies, the tumor percentage of the total amount of biopsies and the maximum percentage of tumor in one cylinder. Univariate (square-Chi, Student t) and multivariate (multiple logistic regression) analysis are performed in order to study the relationship of these parameters with the presence or not of an organ-located disease. RESULTS: An organ-located disease was shown at 44 patients through the piece of radical prostatectomy. The univariate analysis gave all studied parameters, except age, a prognostic value of the existence or not of an organ-located disease. In the multivariate analysis only the total percentage of biopsy tumors (p = 0.0002) and PSA (p = 0.005) behaved as independent prognostic factors. CONCLUSION: Tumor percentage in prostatic biopsy seems to be a factor with a high predictive value of an organ-located disease, possibly because it is an index of tumoral volume.

[Effects of urologic surgery on lymphocyte subpopulations]. / Efectos de la cirugía urológica sobre las subpoblaciones linfocitarias.

We studied the postoperative development of lymphocytes, T cells and lymphocyte subpopulations, in peripheral blood in 45 patients. Twenty-five of them were subjected to a TUR of a bladder tumor, 10 to a transvesical prostatectomy, 6 to a pyelolithotomy and 4 to a nephrectomy for renal atrophy secondary to lithiasis. We confirm a significant postoperative drop in the lymphocyte count and in the percentages of OKT 3+ (total T cells), OKT 4+ (helper T cells) and OKT 8+ cells (cytotoxic/suppressor T cells) 2-4 hours after the operation. Its duration varies according to the type of operation carried out. Despite the slight differences observed in the postoperative development of the lymphocyte subpopulations, at no time in the study did we report significant postoperative variations in the OKT 4+/OKT 8+ quotient.

[Malignant mesothelioma of the tunica vaginalis of the testis]. / Mesotelioma maligno de la túnica vaginal del testículo.

We report a case of malign mesothelioma of the tunica vaginalis of testis in a 84 years-old male. Clinical presentation was hydrocele. Special attention is given to differential diagnosis and therapeuticals possibilities.