Fragmentación del ADN espermático empleando el método de TUNEL / Fragmentation of sperm DNA using the TUNEL method

Objetivos: Establecer la validez del test de TUNEL en la determinación de la fragmentación del ADN espermático, la relación entre el grado de fragmentación con los parámetros seminales y la muestra a emplear para efectuar la prueba. Materiales y métodos: Se emplearon muestras de semen de varones sanos fértiles (n = 33), pacientes que consultaron por infertilidad con prescripción de test de TUNEL (n = 77) y pacientes con fracaso en ICSI (n = 20), analizadas según OMS 2010. Se efectuó el test de TUNEL/ioduro de propidio por citometría de flujo, en muestras basales y post swim up. Resultados: El valor de corte de TUNEL (curvas ROC) fue de 26% con sensibilidad y especificidad de 85 y 89% respectivamente. Las medianas de los resultados de TUNEL pre y post swim up no mostró diferencia significativa (17,0 vs. 12,9%). Sin embargo, el 39,1% expuso una diferencia superior a 15 en valor absoluto entre los resultados del TUNEL basal y post swim up. El estudio de correlación lineal de la morfología, movilidad y vitalidad con el TUNEL post swim up mostró una correlación mayor que el pre selección, con resultados significativos (r: -0,394, p < 0,0001; r: -0,461, p < 0,0001; r: -0,526, p < 0,0001). Conclusiones: La prueba de TUNEL es una prueba válida para su empleo en la clínica. La fragmentación del ADN es un factor independiente de las pruebas tradicionales del semen. Hallamos mayor susceptibilidad al daño generado en los procedimientos de laboratorio en las muestras de peor calidad. La muestra de elección para evaluar la fragmentación del ADN dependerá de si se trata de una fertilización natural o asistida

Objectives: To establish the validity of the TUNEL assay in determining sperm DNA fragmentation, the relationship between the degree of fragmentation and the seminal parameters and the sample needed to conduct the test. Material and methods: We used semen samples from healthy fertile men (n = 33), patients who consulted for infertility with a prescription for the TUNEL assay (n = 77) and patients with intracytoplasmic sperm injection failure (n = 20), analyzed according to the 2010 WHO. The TUNEL/propidium iodide test was performed by flow cytometry, on baseline and post-swim-up samples. Results: The cutoff value for the TUNEL assay (ROC curves) was 26%, with a sensitivity and specificity of 85% and 89%, respectively. The pre-swim-up and post-swim-up medians of the results from the TUNEL assay showed no significant differences (17.0% vs. 12.9%, respectively). However, 39.1% of the samples showed a difference greater than 15 in absolute value between the results of the baseline and post-swim-up TUNEL assays. The linear correlation study of the morphology, mobility and vitality using the post-swim-up TUNEL assay showed a greater correlation than preselection, with significant results (r: -0.394, P < .0001; r: -0.461, P < .0001; r: -0.526, P < .0001). Conclusions: The TUNEL assay is a valid test for clinical use. DNA fragmentation is a factor independent from traditional semen tests. We found a greater susceptibility to damage generated in the laboratory procedures in the samples with lower quality. The sample of choice for evaluating DNA fragmentation will depend on whether the clinician is treating a natural or assisted fertilization

Fragmentation of sperm DNA using the TUNEL method.

OBJECTIVES: To establish the validity of the TUNEL assay in determining sperm DNA fragmentation, the relationship between the degree of fragmentation and the seminal parameters and the sample needed to conduct the test. MATERIAL AND METHODS: We used semen samples from healthy fertile men (n=33), patients who consulted for infertility with a prescription for the TUNEL assay (n=77) and patients with intracytoplasmic sperm injection failure (n=20), analyzed according to the 2010 WHO. The TUNEL/propidium iodide test was performed by flow cytometry, on baseline and post-swim-up samples. RESULTS: The cutoff value for the TUNEL assay (ROC curves) was 26%, with a sensitivity and specificity of 85% and 89%, respectively. The pre-swim-up and post-swim-up medians of the results from the TUNEL assay showed no significant differences (17.0% vs. 12.9%, respectively). However, 39.1% of the samples showed a difference greater than 15 in absolute value between the results of the baseline and post-swim-up TUNEL assays. The linear correlation study of the morphology, mobility and vitality using the post-swim-up TUNEL assay showed a greater correlation than preselection, with significant results (r: -0.394, P<.0001; r: -0.461, P<.0001; r: -0.526, P<.0001). CONCLUSIONS: The TUNEL assay is a valid test for clinical use. DNA fragmentation is a factor independent from traditional semen tests. We found a greater susceptibility to damage generated in the laboratory procedures in the samples with lower quality. The sample of choice for evaluating DNA fragmentation will depend on whether the clinician is treating a natural or assisted fertilization.

Intravascular flow prior to and following angioplasty in peripheral vessels (arteries).

OBJECTIVES: The aim of this study was to assess mean values of velocities in peripheral arterial vessels prior to and following angioplasty in patients with peripheral arterial occlusive disease. Standard values are evaluated with the guidewire positioned proximal to the lesion, inside the lesion and distal to the lesion. METHODS: Twenty-five measurements were taken prior to and following angioplasty in 22 patients with peripheral arterial occlusive disease. The Doppler guidewire was inserted over a 5 French sheath and a Cobra catheter. During measurements, the catheter was totally pulled back and the sheath was placed as proximal as possible. RESULTS: Prior to angioplasty, the velocities in the arteries were 69 +/- 60 cm/second proximal to the lesion, 186 +/- 112 cm/second inside the stenoses and 47 +/- 36 cm/second distal to the lesion. The values increased to 97 +/- 105 cm/second, 89 +/- 89 cm/second and 83 +/- 72 cm/second, respectively. Proximal velocities were lower inside occlusions and increased more markedly compared to stenoses. CONCLUSION: The Doppler guidewire is a practical and valuable tool in assessing technical success after angioplasty of peripheral lesions. In successful angioplasties, no flow acceleration should be observed. Standard values were evaluated.

Energy loss near edge structure of sulfur in non-stoichiometric CuInS(2)

Non-stoichiometric CuInS(2) is a novel material for thin film solar cells. Electron energy loss spectrometry analysis shows that there are considerable lateral inhomogeneities on a scale of less than 100nm. It is as yet unknown whether the Cu or In surplus comes in nanoprecipitates irregularly distributed in the otherwise stoichiometric matrix, or forms supercells where particular atoms are replaced by their complements. This question bears important consequences for the understanding of the local electronic structure and the mechanism of undesired recombination processes in the cell. Since the energy-loss near edge structure of ionisation edges (ELNES) mirrors the local density of unoccupied states it should be different in the two cases. We compare the sulfur L(2,3) ELNES of a Cu-rich and of an In-rich specimen with simulations of supercells based on the TELNES/WIEN97 code. Faint differences in the edge shape can be made visible using difference spectra. They agree reasonably well with simulations. The results show that the surplus species exists as a separate non-stoichiometric phase and not merely as a precipitate of elemental Cu or In.

Intra-arterial Doppler flowmetry in the superficial femoral artery following angioplasty.

The aim of the study was to assess the diagnostic value of an intravascular Doppler guidewire in patients with peripheral percutaneous angioplasty (PTA). The prognostic value was also evaluated. Measurements were done prior and following angioplasty in 22 patients with peripheral arterial occlusive disease. As additional therapy, stent insertion and peripheral (Aa. poplitea Tll/tibial) angioplasty was performed (4 patients per group). For stress testing, adenosinetriphosphate (ATP) was given intra-arterially. Follow-up was performed by angiography, colour-coded duplex ultrasound or judged by unequivocal clinical stage at follow-up to 13 months. Average (APV) and maximal peak velocity (MPV) increased following PTA, after additional treatment (peripheral PTA or stent), and after intra-arterial application of a vasodilator. Patients with peripheral lesions had markedly lower velocities prior treatment and following PTA after vasodilatation. Following peripheral PTA, the values were similar to the patients with PTA alone. Velocities after stenting were markedly increased in the stress condition. Of the 22 patients, 7 had a recurrent disease. The latter patients had higher velocities at rest prior to and following PTA. In stented lesions higher velocities seem to be linked with a worse outcome. The ratio between velocity prior to and after the application of the vasodilator seems to be of diagnostic importance. A ratio of 1.9 or more was of positive prognostic value. The Doppler guidewire is a practical and valuable tool in assessing technical success after angioplasty of peripheral lesions, critical or morphologically worse lesions. In our study the decision for stent application was made on the morphological image; however, increased velocity and changes in phasicity substantiated our decisions. Increased ratios prior to and after vasodilation (flow reserve) are of prognostic value and therefore suitable as indication for stent placement or tibial angioplasty.

[The long-term results after Hydrolyser-supported angioplasty--a prospective study]. / Langzeit-Ergebnisse nach Hydrolyser-unterstützter Angioplastie--eine prospektive studie.

PURPOSE: Data of a three-year follow-up after mechanical thrombolysis with the Hydrolyser catheter were evaluated. Patients have otherwise been treated by local thrombolysis. METHOD: 35 patients were treated by thrombolysis, balloon angioplasty, aspiration, local thrombolysis, and stent placement, if necessary. Morphological results following Hydrolyser treatment and additional treatment were evaluated. RESULTS: Following Hydrolyser treatment a significant reduction of the degree and length of the occlusion was observed. Primary clinical success was 80%. Patency rate after 3 years was 0.5. 23% of all patients died in the follow-up period. CONCLUSION: The Hydrolyser treatment is a relevant alternative to local thrombolysis. This method reduces the time of treatment. The authors favor the lateral opening of the catheter to remove mural thrombus.

[Principles in the assessment of radiation exposure in interventional measures]. / Grundlagen zur Abschätzung der Strahlenexposition bei Interventionellen Massnahmen.

Interventional procedures undoubtedly involve a high radiation exposure, for the patient. For some interventional procedures the frequencies of procedures, radiation exposures and considerations to the radiation risk are shown and discussed. Radiation risk is calculated with the currently common assumptions but always in consideration of the age and expectation of life of the patient. The risk coefficient used for the age-distribution of the common population is 5% per Sv, but for the risk coefficient for, e.g., the group of PTA patients 1.0% per Sv can be derived. That's why the risk of cancer increases in this patient group by 5/100,000. This risk must be balanced with the immediate benefit and the risk of alternative procedures.

When to Stop Angioplasty in Peripheral Vessels (Arteries).

RATIONALE AND OBJECTIVES: The aim of the study was to assess parameters for immediate decisions to optimize peripheral arterial angioplasties and prognostic factors with the help of a intravascular Doppler guide wire in patients with peripheral arterial occlusive disease. METHODS: Measurements were done prior to and following angioplasty in 22 patients with peripheral arterial occlusive disease as additional therapy (stent insertion and tibial angioplasty) was performed. For stress assessment, adenosine-triphosphate (ATP) was given intra-arterially in the vessel periphery. Decisions to perform peripheral angioplasty or stent insertion were made according to morphologic parameters. Follow-up was performed by angiography, color-coded duplex ultrasound or judged by unequivocal improved clinical stage at follow up to 13 month. RESULTS: All but three patients showed an increase in maximal peak velocity (MPV) following angioplasty from 41.2 +/- 23 cm/sec to 67.7 +/- 26 cm/sec. After application of a vasodilator all flow parameters increased in every patient (mean = 61.0 +/- 23.4 cm/sec to 91.0 +/- 32.0 cm/sec). The most well suited parameter to decide stent insertion was the maximal peak velocity in rest. If extremities required peripheral angioplasty the increased flow following ATP application was a valuable parameter. A ratio of 1.9 was of positive prognostic value. CONCLUSIONS: The Doppler guide wire is a practical and valuable tool in assessing technical success after angioplasty of peripheral lesions, critical or tandem lesions. It is suitable for indication to stent insertion or tibial angioplasty and of prognostic value.

Peripheral nerve stimulation by time-varying magnetic fields.

PURPOSE: A study was performed to assess the stimulation threshold for healthy adults using sinusoidally oscillating gradients. METHOD: One hundred thirteen healthy adults were examined in the study. ECG and physiological parameters were measured. All measurements were performed of both the head and the abdomen. The subjects were measured in the supine position with the region of interest positioned in the center of the gradient coils. The measurement was performed for three orthogonal, four oblique, and double oblique orientations. RESULTS: No volunteer reported painful, severe stimulation. The mean thresholds for peripheral stimulation in head and body measurement were similar: 85.5% of stimulation during examination of the head and 87.6% during measurements of the abdomen were reported when the y-gradient was used. CONCLUSION: The greatest frequency of reported stimulations occurs when the y-gradient is used. This was confirmed by the results and supports the hypothesis that orthogonal to the y-axis the body has its largest conductive loop, resulting in the strongest peripheral stimulation.

[Indications and outcome effectiveness of transpopliteal angioplasty]. / Indikation und Erfolg der retrograden transpoplitealen Angioplastie.

PURPOSE: In this retrospective study, the results following angioplasty by the retrograde transpopliteal approach were evaluated to confirm the indications of this technique. PATIENTS AND METHODS: 34 patients were evaluated. Of these 28 were male and 6 were female. Differences to a normal group of patients planned to have angioplasty are the different location of the puncture and on extremely negative selection. RESULTS: Technical success was observed in 84%. A long-term patency ratio of 0.53 was achieved after 30 month. No complications relevant to the therapy were observed. CONCLUSIONS: The transpopliteal approach is a safe and efficient alternative to femoral puncture if the latter is impossible. Compared to the crossover-technique, mechanical considerations favor this technique.

Immediate sealing of arterial puncture site following femoropopliteal angioplasty: a prospective randomized trial.

PURPOSE: A new hemostatic puncture closure device (HPCD) was evaluated following femoropopliteal angioplasty. Efficacy in hemostasis and complications were compared between manual compression and the new system. METHODS: One hundred patients undergoing percutaneous interventional procedures were randomly assigned to receive either manual compression or HPCD. The time to complete hemostasis (when a compression bandage was applied) was noted as well as complications such as hematoma or arterial stenosis at the puncture site. Follow-up was by clinical examination and color-coded duplex sonography (CCDS). RESULTS: With the HPCD, immediate hemostasis was achieved in 22 patients (44%). Discrete oozing without the necessity of external compression or further consequences was observed in 11 patients. Mean manual compression time was 25 (+/- 20) min including application of the pressure bandage. Eleven patients needed additional manual compression and technical failures were observed in 6 patients (12%). The compression time in these 17 cases was 27 (+/- 12) min. Subcutaneous hematomata with a diameter of more than 5 cm developed in 15 of 48 patients in the HPCD group and in 14 of 48 patients in the manual compression group. No surgical or percutaneous intervention was necessary. The complication rate was comparably low in the experimental and control groups. CONCLUSION: Compared with manual compression HPCD is faster and more accurate for sealing the arterial puncture defect following angioplasty. After an initial learning curve, it is easy to handle and time-saving as well as convenient for the patient. Furthermore, immediate and full anticoagulation is possible and arterial inflow is not compromised. A drawback is the necessity of an 8 Fr sheath. Nevertheless, the complication rate is comparably low for both methods.

[Multiplanar imaging in the retroperitoneum: noninvasive diagnosis with a new spiral CT technique--anatomy and pathology]. / Multiplanare Bildgebung im Retroperitoneum: Nichtinvasive Diagnostik mit einer neuen Spiral-CT-Technik--Anatomie und Pathologie.

Both anatomic and pathologic structures are often not documented in sufficient detail to provide overall information in axial scans, and magnetic resonance imaging does not always provide the required spatial resolution. The spiral CT technique, in contrast, with use of the secondary image reconstruction is capable of clearly documenting these interrelations. We have examined 45 patients with pathologic changes in the retroperitoneum by means of a spiral CT scanner and used the secondary image reconstruction in the coronary and sagittal plane in all cases. The scan times ranged between 45 and 60 seconds at a selected slice thickness of 3 mm and a table feed of 4.5 mm. Up to 30 cm of the crania-caudal length of the patients could thus be documented. From the basic data, up to 170 overlapping images were calculated for a multiplanar image reconstruction, and every third axial scan was photographed. The post-processing time was 10 to 15 minutes. The patients received 120 ml of a non-ionic contrast medium, intravenous at a flow rate of 2.5 ml per second. The spiral CT examination was started 40 seconds after the injection had been begun. It could be performed in all patients under inspiration, without problems. The chosen scan times did not lead to significant overshadowing due to motion artefacts. The vascular structures of the aorta and its main branches as well as the vena cava and the portal vein could be identified in their whole lengths. The peritoneal duplications and the renal fascia could be clearly documented. The lymph nodes could be identified in all of them. In cases of tumours with metastases, the extent of the tumor was excellently visible in the multiplanar reconstructions and a thickened peritoneum could be differentiated in such cases. This standardized examination procedure offers a uniform and reproducible diagnostic basis with short examination times. The early detection even of small recurrences in the axial slices and the identification of pathologic changes of the lymph nodes enable a timely tumor treatment for curative purposes.

Electron beam computed tomography in the evaluation of cardiac calcification in chronic dialysis patients.

The purpose of this study was to assess the value of electron beam computed tomography in the detection of cardiac calcifications in coronaries and valves of dialysis patients and to determine the rate at which calcification progresses. Forty-nine chronic hemodialysis patients aged 28 to 74 years were compared with 102 non-dialysis patients aged 32 to 73 years with documented or suspected coronary artery disease, all of whom underwent coronary angiography. We used high-resolution electron beam computed tomography scanning to make 30 axial slices with a distance of 3 mm between each slice. The number of calcifications, the surface area, and the average and highest density values were measured. We calculated a quantitative coronary artery calcium score and assessed calcification of mitral and aortic valves. In dialysis patients, the measurements were repeated after 12 months. The coronary artery calcium score was from 2.5-fold to fivefold higher in the dialysis patients than in the non-dialysis patients. Hypertensive dialysis patients had higher calcium scores than non-hypertensive dialysis patients (P < 0.05). A stepwise, multiple regression analysis confirmed the importance of age and hypertension. No correlation between calcium, phosphate, or parathyroid hormone values and the coronary calcium score was identified; however, the calcium score was inversely correlated with bone mass in the dialysis patients (r = 0.47, P < 0.05). The mitral valve was calcified in 59% of dialysis patients, while the aortic valve was calcified in 55%. The coronary artery calcium score was correlated with aortic valvular, but not mitral valvular calcification. A repeat examination of the dialysis patients at an interval of 1 year showed a disturbing tendency for progression. Our data under-score the frequency and severity of coronary and valvular calcifications in dialysis patients, and illustrate the rapid progression of this calcification. Finally, they draw attention to hypertension as an important risk factor in this process.

[The reduction of the extent of vascular lesions during angioplasty by the use of a sheath]. / Reduktion des Ausmasses von Gefässläsionen im Rahmen der Angioplastie bei Gebrauch einer Schleuse.

PURPOSE: To document the protection of the vessel using an introducer sheath. MATERIAL AND METHODS: In eleven in situ specimens we compared in the inguinal vessel the macroscopic and histologic trauma to the vascular wall during antegrade percutaneous balloon dilatation either without or with a sheath. RESULTS: Significant differences we observed regarding both the puncture site configuration and the puncture hole border, which were evaluated macroscopically. During the histological assessment we found only small intimal trauma in the puncture track in the group where a sheath had been used. Severe intimal damages and the media showed tears if no sheath had been used (100/36% vs. 0/64%). CONCLUSION: It is evident that using a sheath during angioplasty reduces the risk of intimal damage and, consequently, of complications such as recurrent haematoma, stenosis at the puncture site or formation of aneurysms due to the puncture.

[History of interventional radiology]. / Geschichte der interventionellen Radiologie.

Interventional radiology (IR) was only possible following the detection of x-rays by W.C. Röntgen. Also the development of equipments, contrast media, instruments and catheters was necessary. The historical steps since 1964 and some ideas that existed before are discussed, including special highlights in vascular interventional radiology. All the interventional radiological methods used in biliary diseases, urogenital problems and tumour therapy are shortly discussed. The paper gives a personal view of the history of IR, from its beginnings up to day, by a man actively working in this field.

Controversies in vascular imaging and intervention: a panel discussion.

PURPOSE: To express for educational purposes, different and often opposing views on controversial topics of current interest in vascular radiology. And to explore whether or not consensus can be reached on any of these controversial issues. METHOD: Panel discussion among experts in the field. With the help of a moderator, controversial topics were introduced for discussion. For each topic the moderator underscored the principal issue and asked relevant questions. All panelists were asked to express their views on all issues. The following issues were discussed: Atherectomy vs balloon angioplasty; Balloon angioplasty vs stents; Digital vs analog Imaging; TIPS as an emergency procedure; Regional thrombolysis; MRA vs conventional angiography; Laser angioplasty; Carbon dioxide angiography; Lymphangiography vs CT; The training of surgeons in angiography. The panelists were urged to avoid extensive references to the literature but rather to express their own personal opinions based on experience and practice. For each topic an attempt was made to arrive at consensus. RESULTS: Ten issues were presented for debate and discussion. Despite divergent opinions it was possible and relatively painless to reach consensus on seven controversies. On the remaining issues the panelists agreed to disagree. However even when there was disagreement certain trends became apparent. Local considerations and resource availability accounted for varied approaches to the solution of certain controversies. CONCLUSIONS: A panel discussion among experts may be a useful way to address controversies for educational purposes. Despite divergence of opinions consensus may be reached, or in absence of consensus general trends may become apparent.

[Quality, value and risks of pelvic/ leg angiography before PTA (percutaneous transluminal angioplasty)]. / Qualität, Nutzen und Risiken der Becken-/Bein-Angiographie vor PTA.

This study evaluates the diagnostic value and image quality of 183 angiographies performed prior to angioplasty. 56% of the examinations were by arterial and 42% by intravenous DSA. Both techniques were underestimating mild and overestimating severe stenoses. With arterial DSA, 42% of the images in the iliac, 63% in the femoropopliteal, and 53% in the crural region showed an excellent quality. With venous DSA, these percentages were 31%, 47%, and 8%, respectively. Newly occurring occlusions were observed in the iliac region in 8%, and in the femoropopliteal region in 14% of the patients. Venous DSA and other, alternative techniques are sufficient in stage II of POVD and after surgery. In stages III and IV, as well as in diabetic patients and prior to femoro-crural bypasses, an arterial DSA has to be performed. In appropriate lesions, the interval of time to the intervention should be as short as possible to avoid progression of arterial thrombosis.

[Computer program for contrast medium administration in computerized tomography diagnosis]. / Computerprogramme zur Kontrastmittelapplikation in der Computertomographie-Diagnostik.

Contrast medium administration with the contrast medium injector system XD 5500 has proved to be an ideal supplement to the Somatom Plus. The specially developed programs for specific organ regions and differential diagnosis allow an optimization of the diagnosis. The contrast medium administration is thereby standardized and reproducible at any time. Contrast medium enhancement is constant and homogenous with density values of the aorta definitely over 60 Hounsfield units and in most cases over 100 HU. Operation is simple for the proficient investigator and is performed from outside the examination room. The function of a particle filter and an air bubble detector were convincing and certainly ensure the safety of the patient. As a disadvantage, it must be mentioned that the injector carries out program selected even in the event of a perivascular injection. This only occurred three times during our examinations on a total of approximately 5600 patients, without any lasting harm to the patient.