Uterine and Tubal Lavage for Earlier Cancer Detection Using an Innovative Catheter: A Feasibility and Safety Study

Objectives Poor survival of high-grade serous pelvic cancer is caused by a lack of effective screening measures. The detection of exfoliated cells from high-grade serous pelvic cancer, or precursor lesions, is a promising concept for earlier diagnosis. However, collecting those cells in the most efficient way while fulfilling all requirements for a screening approach is a challenge. We introduce a new catheter for uterine and tubal lavage (UtL) and the clinical evaluation of its performance. Methods/Materials In study I, the clinical feasibility of the UtL using the new catheter was examined in 93 patients admitted for gynecologic surgery under general anesthesia. In study II, the safety of the UtL procedure was assessed. The pain during and after the UtL performed under local anesthesia was rated on a visual analog scale by 22 healthy women. Results In study I, the UtL was carried out successfully in 92 (98.9%) of 93 cases by 16 different gynecologists. It was rated as easy to perform in 84.8% of patients but as rather difficult in cancer patients (odds ratio, 5.559;95% confidence interval, 1.434-21.546;P = 0.007). For benign conditions, dilatation before UtL was associated with menopause status (odds ratio, 4.929;95% confidence interval, 1.439-16.884;P = 0.016). In study II, the pain during UtL was rated with a median visual analog scale score of 1.6. During a period of 4 weeks after UtL, none of the participants had to use medication or developed symptoms requiring medical attention. The UtL took 6.5 minutes on average. The amount of extracted DNA was above the lower limit for a sensitive, deep-sequencing mutation analysis in all cases. Conclusions Our studies demonstrate that the UtL, using the new catheter, is a safe, reliable, and well-tolerated procedure, which does not require elaborate training. Therefore, UtL fulfils all prerequisites to be used in a potential screening setting

Nosocomial Outbreak of Parechovirus 3 Infection among Newborns, Austria, 2014

In 2014, sepsis-like illness affected 9 full-term newborns in 1 hospital in Austria. Although results of initial microbiological testing were negative, electron microscopy identified picornavirus. Archived serum samples and feces obtained after discharge were positive by PCR for human parechovirus 3. This infection should be included in differential diagnoses of sepsis-like illness in newborns

Epithelial Ovarian Cancer

Epithelial ovarian cancer generally presents at an advanced stage and is the most common cause of gynaecological cancer death. Treatment requires expert multidisciplinary care. Population-based screening has been ineffective, but new approaches for early diagnosis and prevention that leverage molecular genomics are in development. Initial therapy includes surgery and adjuvant therapy. Epithelial ovarian cancer is composed of distinct histological subtypes with unique genomic characteristics, which are improving the precision and effectiveness of therapy, allowing discovery of predictors of response such as mutations in breast cancer susceptibility genes BRCA1 and BRCA2, and homologous recombination deficiency for DNA damage response pathway inhibitors or resistance (cyclin E1). Rapidly evolving techniques to measure genomic changes in tumour and blood allow for assessment of sensitivity and emergence of resistance to therapy, and might be accurate indicators of residual disease. Recurrence is usually incurable, and patient symptom control and quality of life are key considerations at this stage. Treatments for recurrence have to be designed from a patient's perspective and incorporate meaningful measures of benefit. Urgent progress is needed to develop evidence and consensus-based treatment guidelines for each subgroup, and requires close international cooperation in conducting clinical trials through academic research groups such as the Gynecologic Cancer Intergroup.status: publishe

Ovarian cancer

Ovarian cancer is not a single disease and can be subdivided into at least five different histological subtypes that have different identifiable risk factors, cells of origin, molecular compositions, clinical features and treatments. Ovarian cancer is a global problem, is typically diagnosed at a late stage and has no effective screening strategy. Standard treatments for newly diagnosed cancer consist of cytoreductive surgery and platinum-based chemotherapy. In recurrent cancer, chemotherapy, anti-angiogenic agents and poly(ADP-ribose) polymerase inhibitors are used, and immunological therapies are currently being tested. High-grade serous carcinoma (HGSC) is the most commonly diagnosed form of ovarian cancer and at diagnosis is typically very responsive to platinum-based chemotherapy. However, in addition to the other histologies, HGSCs frequently relapse and become increasingly resistant to chemotherapy. Consequently, understanding the mechanisms underlying platinum resistance and finding ways to overcome them are active areas of study in ovarian cancer. Substantial progress has been made in identifying genes that are associated with a high risk of ovarian cancer (such as BRCA1 and BRCA2), as well as a precursor lesion of HGSC called serous tubal intraepithelial carcinoma, which holds promise for identifying individuals at high risk of developing the disease and for developing prevention strategies

Detection of Müllerian Duct Carcinomas : development and evaluation of a new, innovative diagnostic approach

Weltweit sind mehr als 200,000 Sterbefälle jährlich auf Eierstock- und Endometriumkarzinome zurückzuführen. Beide entwickeln sich aus Müllerschem Epithel und können in zwei Gruppen eingeteilt werden. Typ I wird für gewöhnlich bereits früh diagnostiziert, Type II weist einen wesentlich aggressiveren Phänotyp auf. Type II-Tumore werden auch als „stille Mörder“ bezeichnet, da sie keine, oder nur unspezifische Früh-Symptome verursachen und in den meisten Fällen erst nach der Ausbreitung auf weitere Organe diagnostiziert werden. Die derzeit verwendeten diagnostischen Methoden umfassen die Bestimmung des CA125-Wertes im Serum, sowie ein transvaginaler Ultraschall. Die begrenzte Sensitivität und Spezifität dieser Methoden erlaubt allerdings keine Frühdiagnose der Krebserkrankungen. Diese Arbeit beschreibt die Etablierung einer innovativen Methode zur Detektion von Müllerschen Tumoren. Die Methode basiert auf aktuellen Erkenntnissen, die gezeigt haben, dass Typ II-Tumoren und vielleicht auch Teile von Typ I-Tumoren sich tatsächlich in der Tube entwickeln, wo auch Vorstufen identifiziert wurden. Diese werden als „serous tubal intraepithelial carcinoma“, STIC, bezeichnet. Zellen invasiver Karzinome und STICs werden sehr häufig abgeschilfert. Daraus leitet sich unsere Hypothese ab, wonach diese abgeschilferten Zellen, aufgrund der biologischen Funktion der Tube, in den Uterus transportiert werden. Folglich umfasst unsere Methodik eine Lavage der Uterushöhle um diese Zellen zu gewinnen und in der gewonnenen Spülflüssigkeit nachzuweisen. Der erste Teil der Arbeit beschreibt die Evaluierungsphase. In dieser Phase wurde die Lavage-Methodik, sowie die Weiterverarbeitung der gewonnenen Probe optimiert. Dieser Teil beschreibt auch die erfolgten Maßnahmen, um eine Teilnahme externer Kooperationspartner an weiterführenden Studien zu ermöglichen. Der zweite Teil der vorliegenden Arbeit beschreibt die Ergebnisse, die im Zuge einer proof-of-concept-Studie gewonnen wurden, welche eine Analyse von Lavage-Proben von Ovarialkarzinompatientinnen umfasste. Ein Multiplex-Sequenzierverfahren wurde angewandt, um Mutationen in ausgewählten Genen und somit indirekt Tumorzellen zu detektieren. Bei 18/30 (60 %) der Ovarialkarzinomfälle wurde eine Mutation detektiert. Bei einem Teil der Proben wurde die noch sensitivere Methode - digitale droplet PCR (ddPCR) - angewandt. Dadurch konnte die Anzahl an Ovarialkarzinom-Proben mit IX detektierbarer Mutation auf 24/30 (80 %) erhöht werden. Die Mutationsanteile, die mittels Next generation sequencing und ddPCR bestimmt wurden, zeigten eine Übereinstimmung von R = 0,986 (Spermans Rho). Soweit vorhanden, wurde auch korrespondierendes Tumorgewebe analysiert. Mit Ausnahme von zwei Fällen, konnte dieselbe Mutation, die in der Lavage DNA identifiziert wurde, auch im Gewebe nachgewiesen werden. Dies bestätigt die Gültigkeit unseres Konzepts und somit die Sinnhaftigkeit, dieses auszuweiten und in größeren, spezialisierten Studien anzuwenden. Der dritte Teil der Arbeit beschreibt die ersten Resultate aus diesen laufenden Studien. Im Zuge dieser wurde die Anwendbarkeit des Konzeptes zum Nachweis eines okkulten Ovarialkarzinoms, sowie zur Detektion von 5/5 Endometrium-Karzinomen gezeigt. Des Weiteren wurden 27 Lavage-Proben von Patientinnen mit benignen gynäkologischen Erkrankungen auf das Vorhandensein von Mutationen getestet. In 8/27 (29,6 %) wurde eine Mutation detektiert, wobei in 6/8 (75 %) der Patientinnen das Gen KRAS betroffen war. Diese Arbeit beschreibt die erfolgreiche Etablierung eines neuen Konzeptes zur Detektion von Müllerschen Karzinomen. Die zugrundeliegende Hypothese, wonach Tumorzellen abgeschilfert und anschließend über die Tube in den Uterus transportiert und somit über eine Lavage dieser gewonnen werden können, konnte bestätigt werden. Es wurde gezeigt, dass die Mutationsanalyse ein geeigneter Ansatz ist, um Tumorzellen anschließend in der gewonnen Flüssigkeit zu detektieren. Jedoch ist eine hohe Sensitivität der angewandten Methode erforderlich. Der Grundstein für die Anwendung des Konzeptes im Rahmen größerer Studien konnte gelegt werden. Die ersten darin gewonnenen Resultate waren äußerst vielversprechend, und zeigen das Potential der beschriebenen Methode zur Frühdiagnose von Müllerschen Karzinomen, möglicherweise sogar derer Vorstufen.Ovarian cancer and endometrial cancer account for more than 200,000 deaths each year. Both develop from Müllerian epithelium and can be divided into two groups. Type I usually diagnosed early and type II, which is far more aggressive. Type II cancers are also referred to as the “silent killer”, because they usually only present with unspecific or no early symptoms at all and are frequently not diagnosed before they have already spread. State-of-the-art diagnostic tools are the assessment of the CA125 serum level and a transvaginal ultrasound. However, the sensitivity and specificity of these tests are not high enough to allow for an early detection of these cancers. The present thesis describes an innovative new approach for detecting Müllerian cancers. It is based on recent findings which showed that type II and possibly even type I cancers actually arise in the fallopian tube and that a precursor lesion called “serous tubal intraepithelial carcinoma” (STIC) exists. Cells from invasive carcinoma and already STICs are shed very frequently. We hypothesized that these shed cells are transported into the uterine cavity, based on the natural function of the fallopian tube. Therefore, our approach involves a lavage of the uterine cavity to collect these cells and subsequently detect them in the specimen obtained thereby. The first part of the thesis describes the evaluation phase, including the optimization process of the lavage approach and subsequent processing, as well as different measures which had to be taken to allow conducting the studies within collaborating institutions. The second part describes the results obtained when analyzing samples of ovarian cancer (OC) patients within a proof-of-concept-study. Within this study a massively parallel sequencing approach was used to test the lavage DNA for the presence of mutations in a panel of selected genes. In 18/30 (60 %) of OC cases such a mutation was detected. Additionally, the use of an even more sensitive method for mutation detection was applied in a subset of patients, namely digital droplet PCR (ddPCR). Thereby, the number of positive samples from OC patients could be increased to 24/30 (80 %). The mutation rates obtained by both methods showed a concordance of Spearmans Rho R = 0.986. To the extent available, corresponding tumor tissue DNA was analyzed as well, and in all - except for two cases - the same mutation as in the lavage sample was identified. In this way we were able to confirm the general validity of the concept which in parallel VII indicated that the applicability of this concept should be evaluated within larger, more specific studies. The third part of this thesis describes the first results of these studies, which are still ongoing. In one patient an occult OC could be detected. 5/5 included endometrial cancer cases were tested positive as well. Furthermore, 27 lavage samples of patients with benign gynecological diseases were tested for the presence of a mutation in the same panel of genes. In 8/27 (29.6 %) a mutation was detected. 6/8 (75 %) affected the KRAS gene. This thesis outlines the successful establishment of a new concept for diagnosis of Müllerian cancers. It proves the assumption that tumor cells are shed and can be collected through a lavage of the uterine cavity. Furthermore, it is shown that mutation analysis is a suitable tool for subsequent detection, but a high sensitivity is required. Foundations for larger studies investigating different fields of application of this technique were laid. First promising results were obtained and showed that the described approach has a potential for early diagnosis of Müllerian cancers, possibly even for premalignant lesions.submitted by Elisabeth Maritschnegg, MScZusammenfassung in deutscher SpracheAbweichender Titel laut Übersetzung der Verfasserin/des VerfassersMedizinische Universität Wien, Dissertation, 2016OeBB(VLID)171400

Einfluss der Belastungsdauer bei definierter Intensität auf Ermüdung und Wiederherstellung

Um ein Training optimal auf eine/n Sportler/in anzupassen bedarf es sowohl einer individuellen Gestaltung der Belastungsintensität, als auch der Belastungsdauer. Die „Intensität“ kann man mit dem 3 Phasen-Modell und zwei Turn-Points bereits sehr gut steuern. Hingegen ist in der Praxis ein geeignetes Modell für die „Belastungsdauer“ noch nicht vorhanden und es bedarf weiterer Untersuchungen.Ziel der Studie war es einerseits, den Einfluss der Belastungsdauer in Abhängigkeit zur maximal möglichen Belastungsdauer bei einer definierten Belastungsintensität auf die akute Ermüdung und den Wiederherstellungsverlauf zu überprüfen. ^Andererseits wurden 3 Dauerbelastungen an verschiedenen Belastungsintensitäten über die maximal mögliche Belastungsdauer auf ihren Zusammenhang überprüft.Es handelt sich um eine Einzelfallstudie (Sportstudent: 26 J., 74,5 kg, 1,77 m), daher standen vor allem der methodische Aufbau und die Wirksamkeit der ausgewählten Parameter zur Erfassung der Ermüdung im Vordergrund. Als Belastungen wurden Dauerbelastungen im Laufen mit unterschiedlichen Zeiten abhängig von der maximal möglichen Belastungsdauer vorgegeben. Unterschiedliche Belastungsdauern wurden bei 5%TP2 und Vmax absolviert. Die Ermüdung und Wiederherstellung wurden dabei über HF-Parameter und Sprungtests gemessen.Über die HF-Daten konnte eine belastungsabhängige systemische Ermüdung direkt nach Belastungsende erfasst werden. Mehrere Messzeitpunkte an einem Tag scheinen durch viele Umwelteinflüsse nicht vergleichbar zu sein. ^Die Sprungtests stellten sich nicht als geeigneter Parameter zur Analyse der muskulären Ermüdung heraus.Aus 3 Maximaltests konnte ein linearer Zusammenhang von Geschwindigkeit und Dauer auf einer logarithmischen Zeitachse festgestellt werden.Aus dieser Pilotstudie konnten wichtige methodische Erkenntnisse gewonnen werden. Um dieses Modell in der Praxis anwendbar zu machen, ist weitere Forschung notwendig.To adapt training to an individual athlete both the training-intensity and the training-duration have to be planed individually. The intensity already is described in detail by a 3-phase-model and a 2 turn-point concept. To verify and design an appropriate model we are just in the beginning and further research is necessary.The aim of the study was on the one hand to check the influence of training load depended on the maximal possible training duration a defined training intensity on fatigue and recovery. On the other hand we checked the coherence of 3 constant loads with different training intensities and the maximal possible training duration.It is a single-case-study (sports student: 26 y., 74,5 kg, 177 cm), so especially the methodology and efficacy of the collected parameters to check the fatigue and process of recovery had priority. The workload was a run with constant speed and with different times depended on the maximal possible training duration. 4 training-loads were performed at 5%TP2 and at Vmax. The acute fatigue and recovery were measured by HR parameters and jumping-tests.HR-data allowed to detect fatigue related to the applied duration. Several time points of measures during one day seem to be difficult to compare, because of various environmental imfluences. Jumping-tests didnt expose to be an useful parameter to determine muscular fatigue in this case.From the maximal-tests a linear correlation of speed and duration on a logarithmic time axis was noticed.From this pilot study useful methodological information was collected. To make this model applicable for the practice further research has to be done.vorgelegt von Kevin Maritschnegg, BScAbweichender Titel laut Übersetzung des Verfassers/der VerfasserinZusammenfassungen in Deutsch und EnglischKarl-Franzens-Universität Graz, Masterarbeit, 2017(VLID)231681

Granulomatous Amebic Encephalitis in a Child with Acute Lymphoblastic Leukemia Successfully Treated with Multimodal Antimicrobial Therapy and Hyperbaric Oxygen▿

Acanthamoeba is the causative agent of granulomatous amebic encephalitis, a rare and usually fatal disease. We report a child with acute lymphoblastic leukemia who developed brain abscesses caused by Acanthamoeba during induction therapy. Multimodal antimicrobial chemotherapy and hyperbaric oxygen therapy resulted in complete resolution of symptoms and of pathology as seen by magnetic resonance imaging

KRAS mutation analysis using blood derived cell free tumor DNA for treatment monitoring of metastatic colorectal cancer patients [Poster]

Abstract Currently used non-invasive diagnostic methods for monitoring patients undergoing anti-cancer treatment are limited. The applicability of diagnostic imaging is limited and lack of sensitivity and specificity of routinely used tumor markers hampers clinical application. Circulating cell free DNA (ccfDNA) has been shown to contain minute fractions of tumor-derived DNA. We tested in a proof of principal study the applicability of circulating mutant DNA as a tool to (1) evaluate the clinical sensitivity of cancer detection by analyzing KRAS2 mutations and (2) monitor therapy response in metastatic colorectal cancer (mCRC) patients. In our retrospective study we analyzed 50 serum samples of 18 patients with mCRC and confirmed KRAS2 mutation in the resected primary tumor (G12D, G12V, or G13D). Samples were collected at therapy start (t0), after four cycles of therapy (t1), which was on average after 8 weeks and if available, additional samples at later time points (t2, t3) to assess the longitudinal outcome (median follow up 23 weeks, range 8-248weeks). Therapy response and course of disease were assessed using RECIST criteria at all analyzed time points. The three most commonly occurring KRAS2 (G12D, G12V, or G13D) mutations were examined by Allele-specific PCR. The assay was established using defined tumor cell line DNA and its sensitivity proofed to be up to 0.1%. In 15/18 patients (83.3%), we could detect a KRAS2 mutation at start of therapy (t0) and could confirm diagnosis of mCRC by analysis of ccfDNA. In contrast only 8/18 (44.4%) patients and 7/18 (38.9%) patients showed increased levels of the tumor markers CEA and CA19-9, respectively. Therefore, the analysis of the KRAS2 status identified 5 additional patients as positive for colorectal cancer. To assess therapy response we quantified mutant KRAS2 levels in ccfDNA of 15 patients with detectable KRAS2 mutations. The changes of KRAS2 level were compared to the information from imaging, evaluated by RECIST criteria. A concordance in the assessment of therapy response after 4 cycles (t1) could be observed in 10/15 (66.7%) analyzed cases. In 12/15 patients the longitudinal outcome (t2, t3) was assessed and compared to t1. A concordance in the assessment of therapy response could be observed in 9/12 (75%) patients. Validation of data from patient sample analysis with Deep Sequencing is in progress.Our findings confirm the potential of ccfDNA as a biomarker for improved serum based relapse diagnosis and enables closed meshed monitoring of therapy response. A multiplex mutational analysis will be needed to translate this principle and include also other patient subpopulations (ie. KRAS2 negative). Therefore, we adapted the assay using whole genome amplification to amplify the limited amounts of ccfDNA that now allows the parallel testing of multiple mutations (work in progress). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4513. doi:1538-7445.AM2012-4513</jats:p

Nosocomial Outbreak of Parechovirus 3 Infection among Newborns, Austria, 2014

In 2014, sepsis-like illness affected 9 full-term newborns in 1 hospital in Austria. Although results of initial microbiological testing were negative, electron microscopy identified picornavirus. Archived serum samples and feces obtained after discharge were positive by PCR for human parechovirus 3. This infection should be included in differential diagnoses of sepsis-like illness in newborns