The proteomic analysis of complex body fluids by liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis requires the selection of suitable sample preparation techniques and optimal parameter settings in data analysis software packages to obtain reliable results. Proteomic analysis of follicular fluid, as a representative of a complex body fluid similar to serum or plasma, is difficult as it contains a vast amount of high abundant proteins and a variety of proteins with different concentrations. However, the accessibility of this complex body fluid for LC-MS/MS analysis is an opportunity to gain insights into the status, the composition of fertility-relevant proteins including immunological factors or for the discovery of new diagnostic and prognostic markers for, for example, the treatment of infertility. In this study, we compared different sample preparation methods (FASP, eFASP and in-solution digestion) and three different data analysis software packages (Proteome Discoverer with SEQUEST, Mascot and MaxQuant with Andromeda) combined with semi- and full-tryptic databank search options to obtain a maximum coverage of the follicular fluid proteome. We found that the most comprehensive proteome coverage is achieved by the eFASP sample preparation method using SDS in the initial denaturing step and the SEQUEST-based semi-tryptic data analysis. In conclusion, we have developed a fractionation-free methodical workflow for in depth LC-MS/MS-based analysis for the standardized investigation of human follicle fluid as an important representative of a complex body fluid. Taken together, we were able to identify a total of 1392 proteins in follicular fluid.

In this guideline, recommendations and standards for optimum diagnosis and treatment of endometriosis are presented. They are based on the analysis of the available scientific evidence as published in prospective randomized and retrospective studies as well as in systematic reviews. The guideline working group consisted of experts from Austria, Germany, Switzerland, and the Czech Republic.

In this guideline, recommendations and standards for optimum diagnosis and treatment of endometriosis are presented. They are based on the analysis of the available scientific evidence as published in prospective randomized and retrospective studies as well as in systematic reviews. The guideline working group consisted of experts from Austria, Germany, Switzerland, and the Czech Republic.

In this guideline, recommendations and standards for optimum diagnosis and treatment of endometriosis are presented. They are based on the analysis of the available scientific evidence as published in prospective randomized and retrospective studies as well as in systematic reviews. The guideline working group consisted of experts from Austria, Germany, Switzerland, and the Czech Republic. Zusammenfassung !

3.1 Definition and epidemiology Statements: Endometriosis – one of the most common gynecologic diseases – is defined as the occurence of endometrium-like cell formations outside the uterine cavity. The cardinal symptom is chronic pelvic pain. Infertility is common. There are about 20 000 hospital admissions per year for endometriosis in Germany (Haas et al. 2012). Pathologically and histologically, endometriosis is a benign disease. However, infiltrative growth into adjacent organs is possible requiring extensive surgical procedures.

In this prospective study, we tested the hypothesis if E2 and P serum levels significantly differ during the luteal phase following in vitro-fertilization/intracytoplasmic sperm injection (IVF/ICSI) therapy in conception (CC) versus non-conception (NC) cycles, and their potential in the prediction of pregnancy at the earliest point in time. Serum was sampled from the day of embryo transfer (ET) and throughout the luteal phase until ET + 14 from patients consecutively enrolling for IVF/ICSI therapy. The luteal phase was supported by vaginal P suppositories only, clinical pregnancies were detected by ultrasound and followed up until the 20th week. Overall pregnancy rate was 30.9% constituting the two study groups of CC (n = 22) and NC cycles (n = 49). Significantly, higher E2 (3326 ± 804 versus 1072 ± 233 pmol/l, p = 0.014) and P (244 ± 68 versus 73 ± 10 nmol/l, p = 0.023) were present in CC versus NC from as early as ET + 7. In the CC group, patients with ongoing pregnancies (CC-OG) as compared with miscarriages (CC-MC) had significantly higher E2 and P from ET + 7, predicting ongoing pregnancy in receiver operator characteristics analysis.

German and Austrian Societies for Obstetrics and Gynecology (Deutsche und Österreichische Gesellschaften für Gynäkologie und Geburtshilfe), Association of the Scientific Medical Societies of Germany (Arbeitsgemeinschaft der wissenschaftlichen medizinischen Fachgesellschaften, AWMF).

Im Rahmen der Sterilitätsabklärung wird Endometriose 3–10-mal häufiger beobachtet als in Kollektiven fertiler Frauen. Abgesehen von Zuständen mit offensichtlicher tubarer Komponente ist derzeit unklar, ob die Fertilitätseinschränkung bei unterschiedlichen anatomischen Manifestationen und Ausprägungen der Endometriose auch über unterschiedliche Mechanismen zu erklären ist. Bei Patientinnen mit endometrioseassoziierter Sterilität können das natürliche Fertilitätspotential und einzelne Schritte der Fertilisation in vitro in Abhängigkeit vom beschriebenen Operationssitus analysiert werden, um Mechanismen der Fertilitätseinschränkung verschiedener Endometriosemanifestationen zu evaluieren.

PURPOSE:It has become evident that laparoscopic myomectomy is limited by size, number and location of fibroids. Myomectomy performed by laparotomy can be technically challenging and the surgical benefits have to be weighed against associated risks and impairing fertile potential, especially in multiple and large fibroids that may be positioned close to the cavity. Our aim was to evaluate the effect of microsurgical myomectomy technique on perioperative morbidity in premenopausal women.METHODS:This retrospective study included 228 patients with symptomatic uterine fibroids and/or infertility undergoing myomectomy by laparotomy. As much as 156 patients were treated by standardized microsurgical technique and 72 patients by conventional myomectomy. The following data were recorded and analysed: postoperative haemoglobin, haemoglobin decrease, rate of blood transfusion, and number, size and location of myomas.RESULTS:In 228 patients, seven complications occurred (abdominal wall haematoma, bowel and colon injury, transient ileus). The transfusion rate was 1.3%. Microsurgical technique was associated with a smaller haemoglobin decrease compared to conventional myomectomy (1.77 vs. 2.38 g/dl; P = 0.007). Microsurgical technique correlated inversely with haemoglobin decrease (P < 0.001). Haemoglobin decrease correlated positively with myoma number (P < 0.001), size of myoma (P < 0.001) and the opening of the cavum uteri (P = 0.014).CONCLUSIONS:In this large series of abdominal myomectomies, procedure-related morbidity, mainly perioperative blood loss, was amongst the lowest reported when microsurgical techniques were used. In patients with multiple, large or deep intramural fibroids who desire future pregnancies, the use of microsurgical techniques may decrease intraoperative blood loss and perioperative morbidity.

You have accessJournal of Urology1 Apr 2008PROTAMINE AND Bcl2 mRNA IN TESTICULAR SPERMATIDS AND EJACULATED SPERMATOZOA AS PROGNOSTIC BIOMARKER FOR MALE FERTILITY Klaus Steger, Jochen Wilhelm, Lutz Konrad, Thomas Stalf, Robert Greb, Thorsten Diemer, Sabine Kliesch, Martin Bergmann, and Wolfgang Weidner Klaus StegerKlaus Steger More articles by this author , Jochen WilhelmJochen Wilhelm More articles by this author , Lutz KonradLutz Konrad More articles by this author , Thomas StalfThomas Stalf More articles by this author , Robert GrebRobert Greb More articles by this author , Thorsten DiemerThorsten Diemer More articles by this author , Sabine KlieschSabine Kliesch More articles by this author , Martin BergmannMartin Bergmann More articles by this author , and Wolfgang WeidnerWolfgang Weidner More articles by this author View All Author Informationhttps://doi.org/10.1016/S0022-5347(08)61851-0AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "PROTAMINE AND Bcl2 mRNA IN TESTICULAR SPERMATIDS AND EJACULATED SPERMATOZOA AS PROGNOSTIC BIOMARKER FOR MALE FERTILITY." The Journal of Urology, 179(4S), p. 632 © 2008 by American Urological AssociationFiguresReferencesRelatedDetails Volume 179Issue 4SApril 2008Page: 632 Advertisement Copyright & Permissions© 2008 by American Urological AssociationMetricsAuthor Information Klaus Steger More articles by this author Jochen Wilhelm More articles by this author Lutz Konrad More articles by this author Thomas Stalf More articles by this author Robert Greb More articles by this author Thorsten Diemer More articles by this author Sabine Kliesch More articles by this author Martin Bergmann More articles by this author Wolfgang Weidner More articles by this author Expand All Advertisement PDF DownloadLoading ...

Endometriose und Fertilitätsstörungen: Endometriose reduziert die Fertilität um etwa 50%. In frühen Stadien funktionell in fortgeschrittenen Stadien zusätzlich mechanisch.

Objective To investigate cyclooxygenase (COX-2) expression within different endometriotic lesions and to assess whether these expression patterns correlate with clinical characteristics. Design Retrospective cross-sectional study. Setting University Hospital. Patients Seventy patients with histologically confirmed exclusively peritoneal (n=20), ovarian (n=19) or deep-infiltrating (n=31) endometriosis and a detailed medical history. Intervention Immunohistochemical analysis for COX-2 was performed on 108 endometriotic lesions. Measurements and main results COX-2 intensity, percentage of stained glandular endometriotic cells, and correlation of COX-2 expression with clinicopathological parameters. Semiquantitative COX-2 expression did not differ between distinct morphological types of endometriosis and showed no association with the menstrual cycle. Patients with peritoneal-only endometriosis suffering from moderate or severe chronic pelvic pain showed significantly more frequent COX-2 overexpression than asymptomatic patients or patients with minimal symptoms. In patients with exclusively ovarian or deep-infiltrating endometriosis no association between COX-2 expression and clinical parameters, such as chronic pelvic pain, dysmenorrhoea, dyspareunia, sterility, lower urinary tract symptoms or gastrointestinal symptoms was observed. Conclusion Peritoneal endometriotic lesions with increased COX-2 expression have a special relevance for the development of chronic, nonmenstruation-associated, pelvic pain in endometriotic patients. These patients may benefit from therapy with COX-2 inhibitors.

Angiogenesis plays an important role in the development of the ovarian follicle and its subsequent transition into the corpus luteum. Accordingly, follicular fluid is a rich source of mitogenic and angiogenic factors such as basic fibroblast growth factor and vascular endothelial growth factor secreted by granulosa cells. In the present study, we show that follicular fluid deprived of basic fibroblast growth factor or vascular endothelial growth factor by means of thermal denaturation or antibody neutralization retains its capacity to stimulate endothelial proliferation and angiogenesis. Mass spectrometric analysis of chromatographic fractions stimulating endothelial growth obtained from follicular fluid revealed that the heat-stable mitogenic activity is identical with the subfraction alpha of high density lipoproteins purified from follicular fluid (FF-HDL). Further investigations demonstrated that sphingosine 1-phosphate (S1P), one of the lysophospholipids associated with HDL, accounts for the capacity of this lipoprotein to stimulate endothelial growth and the formation of new vessels. Activation of mitogen-activated protein kinase (p42/44(ERK1/2)), protein kinase C, and protein kinase Akt represent signaling pathways utilized by FF-HDL and S1P to induce endothelial proliferation and angiogenesis. We conclude that FF-HDL represents a novel mitogenic and angiogenic factor present in follicular fluid and that S1P is one of the FF-HDL lipid components accounting for this activity.

The small leucine-rich proteoglycan (SLRP) decorin is efficiently internalized by a variety of cultured cells. A 51-kDa protein has previously been described as a receptor mediating endocytosis of decorin and of the structurally related SLRP biglycan. Recent findings suggest that endocytosis of SLRPs may also be mediated by additional receptors. The class-A scavenger receptor, the endocytic mannose receptor, the epidermal growth factor receptor, and insulin-like growth factor-I receptor have emerged as candidates. We used a combined approach of immunoprecipitation and photoactivated cross-linking to identify endocytosis receptors for decorin in human skin fibroblasts. Decorin was purified by HPLC-DEAE-ion exchange chromatography from the secretions of human skin fibroblasts under nondenaturing conditions. Confocal immunofluorescence microscopy revealed that both biotinylated decorin and decorin conjugated to the heterobifunctional cross-linker sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1-3'-dithiopropionate (SASD) were endocytosed with equal efficiency. SASD-conjugated decorin was added to [35S]-methionine-labeled fibroblasts and cross-linked intracellularly to receptor molecules by photoactivation on endocytic uptake. Cross-linked decorin-receptor complexes were purified from the extracts of trypsin-treated fibroblasts by anion exchange chromatography and immunoprecipitation with a decorin-specific antiserum. Analysis by 2D electrophoresis and autoradiography revealed that decorin was specifically cross-linked to a protein of 110 kDa, which exhibited an isoelectric point of 5.5. In a second approach, unlabeled fibroblasts were subjected to decorin endocytosis and photoactivated cross-linking followed by Western blotting of DEAE-purified cell extracts. A shift of biotinylated decorin immunoreactivity from 165 kDa (decorin-receptor complex) to 54 kDa (SASD-conjugated biotinylated decorin) was noted on reductive cleavage of the cross-linker, representing a difference in molecular weight of approximately 110 kDa. The identification of a 110-kDa protein as a novel endocytosis receptor for decorin provides further support for the emerging concept of a redundancy of receptor molecules in the endocytosis of SLRP.

Das diesjährige Symposion der Deutschen Gesellschaft für Endokrinologie (DGE) vom 9.–12.3. stand ganz im Zeichen der gynäkologischen Endokrinologie. Knapp 600 Teilnehmer folgten der Einladung des Tagungspräsidenten Prof. Ludwig Kiesel nach Münster. Neben der internistischen und pädiatrischen Endokrinologie sollte nach den Vorstellungen des Vorstands der DGE auch die gynäkologische Sichtweise wieder verstärkt in die Gesellschaft integriert werden. Die 10 von den verschiedenen Sektionen der DGE organisierten Plenarsitzungen waren auch mit international renommierten Wissenschaftlern besetzt, wobei sich in diesem Jahr die Reproduktionsendokrinologie wie ein roter Faden durch die verschiedenen Blöcke zog.