In vitro culture of mechanically isolated murine primary follicles in the presence of human platelet lysate PLTMax.

OBJECTIVE: To develop a system for the culture of murine preantral ovarian follicles using Human Serum Albumin (HSA) and Human Platelet Lysate (PLTMax). METHODS: Mechanically isolated preantral follicles (N=146) were obtained from Swiss mice and cultured in DMEM:F12 medium for ten days in a 96-well plate with conical bottom. The medium was supplemented with penicillin, streptomycin, and equine chorionic gonadotropin. Additional proteins were tested in 4 test groups: G1: human serum albumin (HSA), G2: human platelet lysate (PLTM), and G3 and G4: HSA + PLTMax at lower and higher concentrations, respectively. Cellular vitality and oocyte morphology were evaluated on day 11 of culture. RESULTS: The highest follicular growth (3.4 fold) was achieved in HSA (G1), while a significantly lower (1.8 fold) growth was achieved in the presence of PLTM (G2, G4) and even further reduced (1.2 fold) when HSA and PLTM were combined (G3). Cellular vitality was close to 70-80% among the four groups, and the highest number of intact oocytes were found in G1. CONCLUSIONS: PLTM did not improve follicular development and oocyte maturation compared to HSA but preserved cell vitality.

Malacoplakia of the Uterine Cervix: A Case Report.

Malacoplakia is an uncommon chronic granulomatous inflammation that rarely affects the female genital tract. A case of a 78-year-old woman with malacoplakia involving the uterine cervix and the vagina is described. The patient complained of vaginal bleeding. Clinically, a 13-mm mass was detected in the cervix, which was confirmed by ultrasound scan and magnetic resonance imaging. Histological examination showed a dense histiocytic infiltrate with abundant Michaelis-Gutmann bodies involving the uterine cervix and the upper vagina. The presence of Escherichia coli was confirmed in the lesion by immunohistochemistry and polymerase chain reaction. Only 12 cases of cervical malacoplakia have been reported to date. This condition should be included in the differential diagnosis of cervical tumors.

Susceptibility artifacts and PIRADS 3 lesions in prostatic MRI: how often is the dynamic contrast-enhance sequence necessary?

PURPOSE: To assess the need of the dynamic contrast-enhanced (DCE) sequence in addition to T2-weighted imaging (T2-WI) and diffusion-weighted imaging (DWI) for the detection of clinically significant prostate cancer in the presence of artifacts associated with rectal gas (which compromise the diffusion assessment) and/or PIRADS 3 lesions. METHODS: This retrospective study was approved by the institutional review board; informed consent was not required. Patients referred consecutively over a period of 5 months for elevated PSA underwent multiparametric magnetic resonance imaging (mpMRI). mpMRI was performed using a 3T MRI system without an endorectal coil. The MRI findings were reviewed by two radiologists and were scored according to the Prostate Imaging Reporting and Data System version 2.0 (PI-RADSv2). Any discrepancies were resolved by consensus. For statistical purposes, lesions were classified as PIRADS 1-2, PIRADS 3, or PIRADS 4-5. First, all studies were reviewed using a biparametric assessment (T2-WI + DWI), and the presence or absence of susceptibility artifacts was assessed for each prostate. Subsequently, all images were analyzed using the standard multiparametric approach (T2-WI + DWI + DCE). RESULTS: The biparametric evaluation (T2-WI + DWI) showed artifacts (due to the presence of rectal gas or other) in 87 patients (43.5%) and no artifacts in 113 patients (56.5%). In the latter group, 15 patients had peripheral zone (PZ) PIRADS 3 lesions. Thus, a total of 102 patients (51%) had artifacts or PZ PIRADS 3 lesions and therefore required DCE. When evaluating the group of prostates without artifacts, 13.3% of prostates required DCE. A total of 17 (23.9%) PIRADS 4-5 prostate lesions would have not been detected without the use of DCE. CONCLUSION: Biparametric evaluation of the prostate revealed some limitation due to the presence of artifacts or PIRADS 3 PZ lesions. Artifacts were present in almost 44% of our patients, but when the DWI was correctly evaluated, only 13.3% of prostates required DCE.

In vitro maturation of Mus musculus mice oocytes after hyperosmotic shock induced by vitrification solutions.

OBJECTIVE: To evaluate in vitro oocyte maturation rates in embryonic culture medium after induction by hyperosmotic shock caused by exposure to vitrification solutions. METHODS: Bilateral oophorectomy was performed on 20 prepubescent female mice (Swiss). Immature (Prophase I) oocytes (N = 400) were obtained by ovarian dissection, divided into 4 groups, and transferred to culture dishes containing fertilization medium (Sydney IVF Fertilization Medium, Cook® Medical). The control group (CG) did not receive treatment, the test groups (G1, G2, G3) were treated with vitrification solution - 2 (VI-2: 14 M sucrose + ethylene glycol and dimethyl sulfoxide) for 30 seconds and subsequently: G1: 30 seconds in devitrification solution - 2 (DV-2: 0.5M sucrose); G2: 60 seconds DV-2; G3: 60 seconds DV-1(1M sucrose) and 180 seconds DV-2. All groups were cultivated for 24 hours in an incubator at 37ºC and 5% CO2 (Thermo model 3110). After this period, we checked their maturation status. RESULTS: Oocytes exposed to VI-2, DV-1 and DV-2 (G3) showed the highest rate of competence in resuming meiosis and reaching the MII stage; however, there was no statistically significant difference (G3 = 50.5% - 49/97; CG = 27.8% - 10/30). CONCLUSIONS: Oocyte exposure to vitrification solutions, in order to cause osmotic shock, did not interfere with the resumption of meiosis in mice oocytes.

Usefulness of multidetector computed tomography to differentiate between renal cell carcinoma and oncocytoma. A model validation.

OBJECTIVE: The purpose of this study is to validate a multivariable predictive model previously developed to differentiate between renal cell carcinoma (RCC) and oncocytoma using CT parameters. METHODS AND MATERIALS: We included 100 renal lesions with final diagnosis of RCC or oncocytoma studied before surgery with 4-phase multidetector CT (MDCT). We evaluated the characteristics of the tumors and the enhancement patterns at baseline, arterial, nephrographic and excretory MDCT phases. RESULTS: Histopathologically 15 tumors were oncocytomas and 85 RCCs. RCCs were significantly larger (median 4.4 cm vs 2.8 cm, p = 0.006). There were significant differences in nodule attenuation in the excretory phase compared to baseline (median: 31 vs 42, p = 0.015), with RCCs having lower values. Heterogeneous enhancement patterns were also more frequent in RCCs (85.9% vs 60%, p = 0.027).Multivariable analysis showed that the independent predictors of malignancy were the enhancement pattern, with oncocytomas being more homogeneous in the nephrographic phase [Odds Ratio (OR) 0.16 (95% CI 0.03 to 0.75, p = 0.02)], nodule enhancement in the excretory phase compared to baseline, with RCCs showing lower enhancement [OR 0.96 (95% CI 0.93 to 0.99, p = 0.005)], and a size > 4 cm, with RCCs being larger [OR 5.89 (95% CI 1.10 to 31.58), p = 0.038]. CONCLUSION: The multivariable predictive model previously developed which combines different MDCT parameters, including lesion size > 4 cm, lesion enhancement in the excretory phase compared to baseline and enhancement heterogeneity, can be successfully applied to distinguish RCC from oncocytoma. ADVANCES IN KNOWLEDGE: This study confirms that multiparametric assessment using MDCT (including parameters such as size, homogeneity and enhancement differences between the excretory and the baseline phases) can help distinguish between RCCs and oncocytomas. While it is true that this multiparametric predictive model may not always correctly classify renal tumors such as RCC or oncocytoma, it can be used to determine which patients would benefit from pre-surgical biopsy to confirm that the tumor is in fact an oncocytoma, and thereby avoid unnecessary surgical treatments.

Use of Radioactive Seed Localization to Guide Removal of a Nonpalpable Endometriotic Lesion: A Case Report.

Extrapelvic endometriosis is a rare and usually misdiagnosed entity. Some extrapelvic endometriotic lesions are small and nonpalpable, which makes them difficult to locate and remove. Here, we report the use of radioactive seed localization to locate and guide the excision of a small, nonpalpable endometriotic lesion. A 32-year-old woman presented with disabling pain in the right inguinal area. Magnetic resonance imaging and abdominal ultrasound results showed an 11-mm nodule in the abdominal wall, in the vicinity of the groin, consistent with an endometriotic lesion. The radioactive seed was placed within the lesion with the help of ultrasonography, and excision was guided with a portable gamma camera. Complete excision of the endometriotic nodule was achieved. We propose radioactive seed localization as an accurate and feasible technique for the treatment of nonpalpable endometriotic lesions.

Evaluation of the DNA microarray "AMR Direct Flow Chip Kit" for detection of antimicrobial resistance genes from Gram-positive and Gram-negative bacterial isolated colonies / Evaluación de una micromatriz de ADN "AMR Direct Flow Chip Kit" para la detección de genes de resistencia antimicrobiana de bacterias grampositivas y gramnegativas a partir de colonias aisladas

INTRODUCTION: The AMR Direct Flow Chip assay allows the simultaneous detection of a large variety of antibiotic resistance genetic markers. To assess this kit's performance, we use isolated colonies as starting material. The assay has been approved by the European Economic Area as a suitable device for in vitro diagnosis (CE IVD) using clinical specimens. METHODS: A total of 210 bacterial isolates harbouring either one or more antimicrobial resistance genes including plasmid-encoded extended-spectrum β-lactamases (SHV, CTX-M) and carbapenemases (GES, SME, KPC, NMC/IMI, SIM, GIM, SPM, NDM, VIM, IMP, and OXA), mecA, vanA and vanB, and 30 controls were included. RESULTS: The assay displayed a sensitivity and specificity of 100% for all target genes included in the array. CONCLUSION: The AMR Direct Flow Chip Kit is an accurate assay for detecting genes which commonly confer resistance to β-lactams and vancomycin from isolated colonies in culture of Gram-positive and Gram-negative bacteria

INTRODUCCIÓN: El ensayo "AMR Direct Flow Chip Kit" permite detectar simultáneamente la presencia de una gran variedad de marcadores genotípicos de resistencia bacteriana. Evaluamos su rendimiento utilizando colonias aisladas como material de partida. El ensayo aludido ha sido aprobado por el Área Económica Europea como un dispositivo adecuado para el diagnóstico in vitro (CE IVD) utilizando muestras clínicas. MÉTODOS: El estudio ha incluido 210 aislados bacterianos con uno o más genes de resistencia a los antimicrobianos, incluidos genes plasmídicos que codifican β-lactamasas de espectro extendido (SHV y CTX-M) y carbapenemasas (GES, SME, KPC, NMC/IMI, SIM, GIM, SPM, NDM, VIM, IMP y OXA), mecA, vanA y vanB, y 30 controles. RESULTADOS: El ensayo mostró una sensibilidad y especificidad del 100% para todos los genes diana incluidos en la matriz. CONCLUSIÓN: El «AMR Direct Flow Chip Kit» es un ensayo fiable para la detección de genes que comúnmente confieren resistencia a β-lactámicos y vancomicina en bacterias grampositivas y gramnegativas a partir de colonias aisladas en cultivo

Evaluation of the DNA microarray "AMR Direct Flow Chip Kit" for detection of antimicrobial resistance genes from Gram-positive and Gram-negative bacterial isolated colonies.

INTRODUCTION: The AMR Direct Flow Chip assay allows the simultaneous detection of a large variety of antibiotic resistance genetic markers. To assess this kit's performance, we use isolated colonies as starting material. The assay has been approved by the European Economic Area as a suitable device for in vitro diagnosis (CE IVD) using clinical specimens. METHODS: A total of 210 bacterial isolates harbouring either one or more antimicrobial resistance genes including plasmid-encoded extended-spectrum ß-lactamases (SHV, CTX-M) and carbapenemases (GES, SME, KPC, NMC/IMI, SIM, GIM, SPM, NDM, VIM, IMP, and OXA), mecA, vanA and vanB, and 30 controls were included. RESULTS: The assay displayed a sensitivity and specificity of 100% for all target genes included in the array. CONCLUSION: The AMR Direct Flow Chip Kit is an accurate assay for detecting genes which commonly confer resistance to ß-lactams and vancomycin from isolated colonies in culture of Gram-positive and Gram-negative bacteria.

Percutaneous ultrasound-guided vacuum-assisted excision of benign breast lesions: A learning curve to assess outcomes.

OBJECTIVE:: To evaluate the efficacy and learning curve of ultrasoundguided vacuum-assisted excision (US-VAE) of benign breast lesions, and to assess characteristics associated with residual lesion. METHODS:: This was a retrospective study with institutional review board-approval. Sonographic and clinical follow-up were performed 6 months after intervention. Effectiveness and safety of the technique were analyzed. The cumulative summation (CUSUM) graphs were used to evaluate learning curves concerning complete excision and hematoma. RESULTS:: 152 ultrasound-VAEs in 143 patients were included. Initial complete resection was achieved in 90.8 % (138 of 152). 6-month follow-up was completed for 143 (94%) of cases and complete resection was observed in 72 % (100 of 143). Mean maximum size without residual tumor was 16.9 mm, while with residual lesion it was 21.9 mm (p = < 0.001), with a volume of 1.53 and 3.39 cm3, respectively (p = < 0.001). Increase in lesion size and volume was associated with less effectiveness (p = 0.05), clinical control (p = 0.05), and higher risk of clinically significant hematoma (p = 0.05). Receiver operating characteristic analysis demonstrate a volume threshold of 2.6 cm3 (r = 0.71, specificity 84.5%) for leaving no residual lesion. Cumulative summation graphs demonstrate that, on average, 11 excisions were required to acquire skills to perform complete excision in more than 80% at the end of the ultrasound-VAE and 18 excisions at 6 months. CONCLUSION:: Ultrasound-VAE is an effective treatment for benign breast lesions. Breast lesion volume should be considered when assessing for percutaneous treatment. ADVANCES IN KNOWLEDGE:: A follow-up of the learning process of ultrasound-VAE will be a valuable tool to assess the efectiveness and safety of the technique i.

Wistar rats immature testicular tissue vitrification and heterotopic grafting.

OBJECTIVE: To evaluate the efficiency of two vitrification protocols for rat immature testicular tissue and heterotopic transplantation. METHODS: Twenty-four pre-pubertal Wistar rats were divided into three groups (n=8). After orchiectomy, testicular fragments (3mm) from Groups 1 and 2 were vitrified with different cryoprotectant concentration solutions, using sterile inoculation loops as support. After warming up, the fragments were submitted to cell viability assessment by Trypan blue and histological evaluation. Vitrified (Groups 1 and 2) and fresh (Group 3) fragments were grafted to the animals periauricular region. After 8 weeks of grafting, the implant site was histologically analyzed. RESULTS: The viability recovery rate from Group 1 (72.09%) was higher (p=0.02) than that from Group 2 (59.19%). Histological analysis showed similar tubular integrity between fresh fragments from Groups 1 and 3. Group 2 samples presented lower tubular integrity. We ran histological analyses in the grafts from the Groups. In all groups, it was possible to see the implant site, however, no fragment of testicular tissue or signs of inflammation were histologically found in most samples from Groups 1 and 3. In one sample from Group 2, we found degenerated seminiferous tubules with necrosis and signs of an inflammatory process. In another sample from Group 2, we found seminiferous tubules in the implant site. CONCLUSION: The vitrification of pre-pubertal testicular tissue of rats showed little damage to cell viability through histological analysis when we used cryoprotectants in a lower concentration. Heterotopic transplantation could not preserve the structural organization of the testicular tissue.

Ovarian tissue vitrification and heterotopic autologous transplantation in prepubertal Wistar rats.

OBJECTIVE: To evaluate the efficiency of ovarian tissue heterotopic autografting after vitrification in prepubertal rats. METHODS: Fragments of excised ovaries from prepubertal rats were used after assessing post-warming cellular viability, to determine the best vitrification protocol prior to retroauricular autografting. Pre-pubertal females (N=24) were castrated and divided into three group: Group 1 - fresh ovarian tissue transplantation; Group 2 - vitrified/warmed tissue transplantation; Group 3 - bilateral oophorectomy without transplantation. The ovarian fragments were exposed to solutions from the Ingamed® commercial kit, allocated in bacteriological loops and immersed in liquid nitrogen. Sixty days after transplantation, a vaginal mucus sample was collected for cytology tests, followed by sacrificing the animal, performing a cardiac puncture for collecting a blood sample to determine luteinizing hormone and estradiol levels, and excision of the transplanted fragment for histology tests. RESULTS: Vaginal cytology revealed that 87.5% of females from groups 1 and 2 had estrus while all females in Group 3 remained in diestrus. The mean LH value in groups 1 (0.08 mIU/mL) and 2 (0.34 mIU/mL) were statistically different from that of Group 3 (2.27 mIU/mL). E2 values did not differ between the groups. The histological analysis of Group 1 excised grafts versus those from Group 2 showed a higher percentage of primary follicles (62.5% vs. 12.5%), developing follicles (75% vs. 25%), corpus luteum (37.5% vs. 12.5%) and stromal region (100% vs. 87.5%). CONCLUSION: This study indicated that pre-pubertal ovarian tissue vitrification can be used to preserve fertility and to restore endocrine function in castrated rats.

Could cryopreserved human semen samples be stored at -80°C?

OBJECTIVE: To evaluate storage time effects in cryopreserved human semen samples, kept in the freezer at a controlled temperature of -80°C, on sperm viability after thawing. METHODS: We used 20 semen samples. Each sample was cryopreserved in 10 fingers, which were divided into five groups: one group was kept in cryogenic canisters throughout the experiment(control), and four groups were kept in a VIP Ultra Low MDF-U76V- PE freezer, with the temperature set at -80°C, for 24, 48, 72 and 96 hours, respectively. After the exposure time, the samples were stored in cryogenic canisters after being thawed. The analyzed parameters were: motility, vitality and mitochondrial activity. RESULTS: After thawing, we noticed decreased sperm motility, vitality and mitochondrial activity, when comparing the tested groups with the control group, as well as a progressive reduction in the analyzed parameters between the times evaluated. CONCLUSIONS: Cryopreservation of semen samples at -80°C is potentially harmful to sperm viability, causing damage when submitted to longer exposure times.

Effects of energy drinks on biochemical and sperm parameters in Wistar rats

BACKGROUND: The present study evaluates the effects of energy drinks on the reproductive and biochemical parameters of adult male rats. METHODS: A total of 40 male rats (Wistar) were exposed to an energy drink mixed with the drinking water for a period of 120 days. The animals were divided into four groups and exposed to increasing therapeutic doses (DT) of an energy drink, based on allometric extrapolation, resulting in values (mL/day) per animal of 250 g: DT1 2.36 mL, DT3 7.47 mL, and DT6 14.16 mL. The control group (CTRL) consumed water only. During the treatment, the rats were assessed for signs of toxicity. After treatment, the animals were sacrificed and their organs were weighed. Sperm parameters (motility, concentration, and morphology) were evaluated. The biochemical markers alanine eamino transferase, aspartate amino transferase, alkaline phosphatase, lactic dehydrogenase, urea, creatinine, creatine phosphokinase, and creatine kinase MB fraction were measured, in addition to total cholesterol and testosterone. RESULTS: There was a significant decrease (p< 0.05) in the concentration of sperm in the treated groups (DT18.5 ± 0.7; DT3 7.2 ± 0.9; DT6 8.4 ± 0.9) compared to the control group (12.3 ± 1.2). No difference was observed with respect to relative weights of the animals'organs, water consumption, signs of toxicity, behavioral changes, biochemical markers, and sperm motility and morphology. CONCLUSION: The long-term consumption of energy drinks interferes negatively with sperm concentration, without affecting sperm motility and morphology or altering the hepatic, cardiac, or renal functions

Endometrial Stromal Sarcoma Arising in Colorectal Endometriosis.

The malignant transformation of endometriosis is very uncommon. Whereas 75% of tumors arising from endometriosis arise in the ovary, location in extra-genital organs is rare and mesenchymal neoplasms are exceptional. A 47 year-old woman who underwent hysterectomy with bilateral salpingo-ooforectomy due to endometriosis 13 years before presented with abdominal pain. The magnetic resonance imaging (MRI) showed a 9.7×7.5 cm solid-cystic supravesical mass and a recto-vaginal tumor, as well as endometriotic nodules in the sigma, right parametrium and peritoneum that had significantly increased in size over a six months period. The patient underwent surgical resection of the masses. The histological study showed a low-grade endometrial stromal sarcoma (ESS) arising from endometriosis located at recotovaginal septum and affecting colonic wall and multiple peritoneal and pelvic implants. The patient received radiotherapy and aromatase inhibitors and is free of disease after a follow up of 2 years. Only 15 cases of ESS arising in endometriosis of the bowel have been reported. Tumor dissemination at diagnosis is unusual but does not imply a poor prognosis, as only one patient has died due to progression of the tumor. ESS should be included in the differential diagnosis of mesenchymal neoplasms in the intestine.

Accuracy of unenhanced magnetic resonance angiography for the assessment of renal artery stenosis.

PURPOSE: To evaluate the accuracy of unenhanced magnetic resonance angiography (U-MRA) using balanced steady-state free precession (SSFP) sequences with inversion recovery (IR) pulses for the evaluation of renal artery stenosis. MATERIALS AND METHODS: U-MRA was performed in 24 patients with suspected main renal artery stenosis. Two radiologists evaluated the quality of the imaging studies and the ability of U-MRA to identify hemodynamically significant main renal artery stenosis (RAS) defined as a stenosis ≥50% when compared to gold standard tests: contrast-enhanced magnetic resonance angiography (CE-MRA) (18 patients) or digital subtraction arteriography (DSA) (6 patients). RESULTS: A total of 44 main renal arteries were evaluated. Of them, 32 renal arteries could be assessed with U-MRA. When CE-MRA or DSA was used as the reference standard, nine renal arteries had hemodynamically significant RAS. U-MRA correctly identified eight out of nine arteries as having ≥50% RAS, and correctly identified 22 out of 23 arteries as not having significant RAS, with a sensitivity of 88.8%, a specificity of 95.65%, positive and negative predictive value of 88.8% and 95.65%, respectively, and an accuracy of 93.75%. Renal artery fibromuscular dysplasia (FMD) was observed in the two misclassified arteries. CONCLUSION: U-MRA is a reliable diagnostic method to depict normal and stenotic main renal arteries. U-MRA can be used as an alternative to contrast-enhanced magnetic resonance angiography or computer tomography angiography in patients with renal insufficiency unless FMD is suspected.

Usefulness of MDCT to Differentiate Between Renal Cell Carcinoma and Oncocytoma: Development of a Predictive Model.

OBJECTIVE: The objective of our study was to identify the most useful parameters to differentiate between renal cell carcinoma (RCC) and oncocytoma using four-phase CT. MATERIALS AND METHODS: Ninety-seven patients with solid renal lesions who underwent surgery with four-phase preoperative CT evaluation and with pathologic diagnosis of RCC or oncocytoma were included in the study. Features of tumors and the enhancement pattern in the four CT phases were evaluated and analyzed. Logistic regression models were used to assess independent predictors for malignancy. RESULTS: Histopathologically, 13 tumors were oncocytomas and 84 were RCCs. RCCs were larger (6.20 cm vs 3.21 cm, p = 0.0004) and more often enhanced heterogeneously (66 vs 6, p = 0.02). Lesions that were larger than 4 cm showed a significantly higher risk of malignancy (p = 0.0046). Significant differences were found in intensity of nodule enhancement between the nephrographic and the excretory phases with respect to the unenhanced phase (p = 0.003 and p = 0.0026). At multivariate analysis, parameters that were independent predictors of malignancy were enhancement pattern, with RCCs more often having heterogeneous enhancement than oncocytomas (odds ratio [OR], 0.18; 95% CI, 0.04-0.90), and nodule enhancement in the excretory phase in relation to the unenhanced phase, with RCCs showing lower enhancement (OR, 0.93; 95% CI, 0.88-0.97), and a size larger than 4 cm (OR, 4.01; 95% CI, 0.70-23.14). CONCLUSION: The combination of different CT parameters including lesion size larger than 4 cm, lesion enhancement in the excretory phase in relation to the unenhanced phase, and heterogeneous enhancement pattern helps distinguish RCC from oncocytoma.

The effects of storing and transporting cryopreserved semen samples on dry ice.

OBJECTIVE: This study aimed to test the effects on sperm viability of transporting cryopreserved semen samples on dry ice. METHODS: Twenty normozoospermic semen samples were cryopreserved and divided into five groups. The samples in Group 1 were immersed in liquid nitrogen throughout the experiment in cryogenic storage tanks; the cryopreserved straws in Group 2 were placed in a Styrofoam box containing dry ice and kept under these conditions for 48 hours; the samples in Group 3 were kept for 48 hours on dry ice under the same conditions as the Group 2 samples, and were then moved to a storage tank filled with liquid nitrogen; Group 4 samples were also kept for 48 hours in dry ice storage, and the Styrofoam box containing the samples was shipped by plane to assess the effects of shipping; the samples in Group 5 were shipped together with the Group 4 samples and were placed in a storage tank with liquid nitrogen after spending 48 hours stored on dry ice. After thawing, sperm parameters were analyzed for viability, vitality, and motility; spermatozoa were also tested for mitochondrial activity. RESULTS: Significant decreases in motility recovery rates (P=0.01) and vitality (P=0.001) were observed in all groups when compared to the control group. Mitochondrial activity was significantly decreased only in Group 5 (P=0.04), as evidenced by greater numbers of sperm cells not stained by reagent 3,3'-diaminobenzidine. CONCLUSIONS: Transportation did not affect the quality of cryopreserved semen samples, but dry ice as a means to preserve the samples during transportation had detrimental effects upon the sperm parameters assessed in this study.

Pathways of lymphatic spread in gynecologic malignancies.

Precise radiologic evaluation of regional adenopathic involvement in pelvic gynecologic tumors is fundamental to clinical practice because of its prognostic and therapeutic significance. Likewise, the identification of metastatic adenopathies at posttreatment imaging is essential for assessing response and detecting recurrence. Similar to urologic neoplasms, gynecologic neoplasms most often spread regionally to the pelvic and retroperitoneal lymph nodes, following the normal drainage pathways of the pelvic organs. Familiarity with routes of dissemination, treatment options, and means of analyzing lymph node characteristics is crucial to determine the extent of disease. Two staging systems can be used in characterizing gynecologic malignancies: the FIGO (International Federation of Gynecology and Obstetrics) system, which is the most commonly and universally used, and the TNM (tumor, node, metastasis) system, which is based on clinical and/or pathologic classification. Anatomic assessment with multidetector computed tomography (CT) and magnetic resonance (MR) imaging is still the most commonly used technique for the detection of lymph node spread, which is mainly based on morphologic criteria, the most important of which is nodal size. However, size has limited diagnostic specificity. Consequently, functional imaging techniques such as diffusion-weighted MR imaging, positron emission tomography combined with CT, lymphoscintigraphy, and sentinel lymph node mapping, which are based on molecular and physiologic activity and allow more precise evaluation, are often incorporated into diagnostic imaging protocols for staging of gynecologic malignancies.

Sperm Selection Using Three Semen Processing Techniques.

OBJECTIVE: This study aimed to assess the efficiency, in terms of recovered motile spermatozoa with normal morphology, of three sperm selection techniques: migration- sedimentation (SS), swim-up from fresh semen (SF), and swim-up from washed (SL) sperm. METHODS: Samples from 20 normozoospermic men were divided into three equal aliquots and processed in parallel. SS was performed in a Jondet tube, using 1 ml of semen and 2.5 ml of Human Tubal Fluid medium (HTF+10% Synthetic Serum Supplement, Irvine, USA). For SF, 1 ml of HTF was layered over 1 ml of fresh semen (SF). For SL, 1 ml of sperm was first centrifuged (300 g, 10 min) and the pellet resuspended in 1 ml of HTF; a second layer of HTF was placed on top. Migration time was 1h (SF and SL) and 1h30' for SS at 37°C. After migration, 200 µl were removed from the top layer (SF, SL) and from the central cone (SS). Concentration, morphology and motility were determined. RESULTS: Recovery rates were 25% for SS, 10.1% for SF and 4.5% for SL. SS recovery rate was significantly higher (P<0.01) than the two swim-up techniques. Total motility was statistically different (P<0.001), with 93.6% for SS, 91.2% for SF, and 77% for SL. Sperm morphology was similar between the three techniques (P= 0.12). CONCLUSION: SS is an efficient technique for the recovery of motile spermatozoa from native semen preparations and yielded better results than SF and SL. Routine use for assisted reproduction remains to be evaluated.

Prospective evaluation of CT indeterminate renal masses using US and contrast-enhanced ultrasound.

OBJECTIVE: The aim of this study was to assess the accuracy of Ultrasound (US) and contrast-enhanced ultrasound (CEUS) in the characterization of renal nodules indeterminate on CT by identifying benign cystic lesions not requiring further examination. METHODS: 72 patients with 83 indeterminate renal nodules on CT underwent baseline US and CEUS that classified lesions as benign (Bosniak I, II or IIF cysts) or potentially malignant (Bosniak III or IV cysts, solid nodules). The accuracy of US and CEUS in the differentiation between benign cysts and potentially malignant nodules was analyzed and compared with the final diagnosis obtained by histology or follow-up of at least 23 months with CEUS ± a conclusive CT/MR study. RESULTS: Final diagnoses comprised 50 benign complex cysts, 1 focal nephritis, 1 multilocular cystic nephroma, 3 oncocytomas, 1 transitional cell carcinoma and 27 renal cell carcinomas. Unenhanced US correctly classified 18/50 (36%) benign cysts and 17/33 (51.5%) of the potentially malignant lesions obtaining a sensitivity of 36%, specificity of 51.5%, and overall accuracy of 42.2%. The addition of CEUS allowed a correct diagnosis of 48 /50 (96%) benign cysts and of 31/33 (93.9%) nodules as potentially malignant, with a sensitivity of 96%, specificity of 93.9%, and overall accuracy of 95.2%. CONCLUSION: CEUS is very useful in the differentiation between benign complex cysts and other lesions that require further investigation in non-conclusive renal nodules detected on CT, improving the accuracy of baseline US from 42.2 to 95.2%.