Doerthe Brueggmann

Deficiencies in immunoassay methods used to monitor serum Estradiol levels during aromatase inhibitor treatment in postmenopausal breast cancer patients

Optimal care for breast cancer patients undergoing aromatase inhibitor (AI) treatment is ensured when estradiol (E2) levels are adequately suppressed. To assess treatment efficacy accurately, it is important to measure the serum E2 levels using a well validated assay method with high sensitivity and specificity. This translates into the urgent need to evaluate various E2 immunoassay kits, which are frequently used in hospital settings to measure E2 serum levels in patients undergoing AI treatment, so clinicians obtain accurate and reliable measurements allowing appropriate clinical decision making. Our objective was to evaluate the performance of different commercially available and commonly used E2 immunoassay kits regarding measurement of E2 levels in the serum of postmenopausal breast cancer patients treated with AIs, in comparison to a highly accurate and reliable mass spectrometry assay. Clinical and demographic data were obtained from 77 postmenopausal breast cancer patients who were treated with an AI. Serum E2 levels were measured by 6 immunoassay methods and by liquid chromatography-tandem mass spectrometry (LC-MS/MS), which served as the standard for comparison. Analysis of E2 by LC-MS/MS showed that 70% of the samples had levels that were <5 pg/ml. Three of the assays carried out with commercial E2 immunoassay kits had poor sensitivities and were not able to detect E2 levels <10 or <20 pg/ml. Although two of the E2 assays using commercial kits demonstrated a better sensitivity (5 pg/ml), the measured E2 values were substantially higher than those obtained by LC-MS/MS. The assay with the sixth commercial E2 kit grossly underestimated the true E2 values. E2 assays carried out with commercial E2 immunoassay kits lack the accuracy to measure the very low serum E2 levels found in patients being treated with AIs. Serum samples from such patients should be sent to laboratories that use a mass spectrometry assay.

Novel three-dimensional in vitro models of ovarian endometriosis.

BackgroundEndometriosis is characterized by the presence of functional endometrial tissue outside of the uterine cavity. It affects 1 in 10 women of reproductive age. This chronic condition commonly leads to consequences such as pelvic pain, dysmenorrhea, infertility and an elevated risk of epithelial ovarian cancer. Despite the prevalence of endometriosis and its impact on women’s lives, there are relatively few in vitro and in vivo models available for studying the complex disease biology, pathophysiology, and for use in the preclinical development of novel therapies. The goal of this study was to develop a novel three-dimensional (3D) cell culture model of ovarian endometriosis and to test whether it is more reflective of endometriosis biology than traditional two dimensional (2D) monolayer cultures.MethodsA novel ovarian endometriosis epithelial cell line (EEC16) was isolated from a 34-year old female with severe endometriosis. After characterization of cells using in vitro assays, western blotting and RNA-sequencing, this cell line and a second, already well characterized endometriosis cell line, EEC12Z, were established as in vitro 3D spheroid models. We compared biological features of 3D spheroids to 2D cultures and human endometriosis lesions using immunohistochemistry and real-time semi-quantitative PCR.ResultsIn comparison to normal ovarian epithelial cells, EEC16 displayed features of neoplastic transformation in in vitro assays. When cultured in 3D, EEC16 and EEC12Z showed differential expression of endometriosis-associated genes compared to 2D monolayer cultures, and more closely mimicked the molecular and histological features of human endometriosis lesions.ConclusionsTo our knowledge, this represents the first report of an in vitro spheroid model of endometriosis. 3D endometriosis models represent valuable experimental tools for studying EEC biology and the development of novel therapeutic approaches.

Density equalizing mapping of the global tuberculosis research architecture

BACKGROUND Tuberculosis belongs to the lung infectious diseases with the highest impact on global burden of disease. Yet there is no concise scientometric study about tuberculosis research. Therefore, the NewQiS project elected this subject as focus of an in depth analysis to perform density equalizing mapping in combination with scientometrics. METHOD In this retrospective study all publications related to tuberculosis research listed in the Web of Science database between 1900 and 2012 were identified, analyzed and submitted to density equalizing mapping procedures. RESULTS In total 58,319 entries on TBC were identified with the USA being the most productive country with 11,788 publications, followed by the United Kingdom (4202), India (3456), France (2541), South Africa (1840), Germany (1747) and China (1427). Concerning the citations rate Denmark leads with 43.7 citations per article, followed by Latvia (39.1), Gambia (38.3), Senegal (34.9), and the Netherlands (31.4). Chart techniques demonstrates a widely ramified international network with a focus the joint work of USA, the UK and South Africa. CONCLUSIONS This is the first density equalizing and scientometric study that addresses tuberculosis research over a period of 112 years. It illustrates global tuberculosis research architecture and stresses the need for strengthening global research efforts and funding program.

Gender disparities in high-quality research revealed by Nature Index journals

Background The present study aims to elucidate the state of gender equality in high-quality research by analyzing the representation of female authorships in the last decade (from 2008 to 2016). Methods Based on the Gendermetrics platform, 293,557 research articles from 54 journals listed in the Nature Index were considered covering the categories Life Science, Multidisciplinary, Earth & Environmental and Chemistry. The core method was the combined analysis of the proportion of female authorships and the female-to-male odds ratio for first, co- and last authorships. The distribution of prestigious authorships was measured by the Prestige Index. Results 29.8% of all authorships and 33.1% of the first, 31.8% of the co- and 18.1% of the last authorships were held by women. The corresponding female-to-male odds ratio is 1.19 (CI: 1.18–1.20) for first, 1.35 (CI: 1.34–1.36) for co- and 0.47 (CI: 0.46–0.48) for last authorships. Women are underrepresented at prestigious authorships compared to men (Prestige Index = -0.42). The underrepresentation accentuates in highly competitive articles attracting the highest citation rates, namely, articles with many authors and articles that were published in highest-impact journals. More specifically, a large negative correlation between the 5-Year-Impact-Factor of a journal and the female representation at prestigious authorships was revealed (r(52) = -.63, P < .001). Women publish fewer articles compared to men (39.0% female authors are responsible for 29.8% of all authorships) and are underrepresented at productivity levels of more than 2 articles per author. Articles with female key authors are less frequently cited than articles with male key authors. The gender-specific differences in citation rates increase the more authors contribute to an article. Distinct differences at the journal, journal category, continent and country level were revealed. The prognosis for the next decades forecast a very slow harmonization of authorships odds between the two genders.

Differential gene expression by 1,25(OH)2D3 in an endometriosis stromal cell line

Endometriosis is a common female reproductive disease characterized by invasion of endometrial cells into other organs, frequently causing pelvic pain and infertility. Alterations of the vitamin D system have been linked to endometriosis incidence and severity. To shed light on the potential mechanism for these associations, we examined the effects of 1,25(OH)2D3 on gene expression in endometriosis cells. Stromal cell lines derived from endometriosis tissue were treated with 1,25(OH)2D3, and RNA-seq was used to identify genes differentially expressed between treated and untreated cells. Gene ontology and pathway analyses were carried out using Partek Flow and Ingenuity software suites, respectively. We identified 1627 genes that were differentially expressed (886 down-regulated and 741 up-regulated) by 1,25(OH)2D3. Only one gene, CYP24A1, was strongly up-regulated (369-fold). Many genes were strongly down-regulated. 1,25(OH)2D3 treatment down-regulated several genetic pathways related to neuroangiogenesis, cellular motility, and invasion, including pathways for axonal guidance, Rho GDP signaling, and matrix metalloprotease inhibition. These findings support a role for vitamin D in the pathophysiology of endometriosis, and provide new targets for investigation into possible causes and treatments.

Expression of Wnt signaling pathway genes in human endometriosis tissue: a pilot study

Despite the high prevalence of endometriosis, little is known about the underlying disease mechanisms [1]. The development and maintenance of endometriosis lesions are linked to the process of epithelial–mesenchymal transition (EMT), which describes the transformation of stationary epithelial cells into proliferative and invasive mesenchymal cells [2]. Since the canonical Wnt-signaling system is an important driver for EMT [3], we hypothesized that Wnt-signaling might be altered in different types of endometriosis lesions [4] leading to changes in related EMT processes. Hence, this study’s objective was to quantify the expression changes of (1) Wnt-signaling pathway genes such as Wnt-agonists, antagonists and receptors and (2) Wnt-target genes (e.g. E-cadherin and S100A4), which play a role in EMT, in endometrium from women with endometriosis (cases) and without (controls) as well as endometriosis lesions at different stages of the cycle.

Src as a novel therapeutic target for endometriosis.

BACKGROUND Endometriosis is a common condition that is associated with an increased risk of developing ovarian carcinoma. Improved in vitro models of this disease are needed to better understand how endometriosis, a benign disease, can undergo neoplastic transformation, and for the development of novel treatment strategies to prevent this progression. METHODS We describe the generation and in vitro characterization of novel TERT immortalized ovarian endometriosis epithelial cell lines (EEC16-TERT). RESULTS Expression of TERT alone was sufficient to immortalize endometriosis epithelial cells. TERT immortalization induces an epithelial-to-mesenchymal transition and perturbation in the expression of genes involved in the development of ovarian cancer. EEC16-TERT was non-tumorigenic when xenografted into immunocompromised mice but grew in anchorage-independent growth assays in an epidermal growth factor and hydrocortisone dependent manner. Colony formation in agar was abolished by inhibition of Src, and the Src pathway was found to be activated in human endometriosis lesions. CONCLUSIONS This new in vitro model system mimics endometriosis and the early stages of neoplastic transformation in the development of endometriosis associated ovarian cancer. We demonstrate the potential clinical relevance of this model by identifying Src activation as a novel pathway in endometriosis that could be targeted therapeutically, perhaps as a novel strategy to manage endometriosis clinically, or to prevent the development of endometriosis-associated ovarian cancer.

Perceptions and patterns of cervical cancer screening among predominately Hispanic women who have sex with women: a cross-sectional study

The major risk factor for cervical cancer – the Human Papilloma Virus (HPV) – is identified in up to 30% of women having sex with women (WSW) [1]. WSW undergo Papanicolaou screening (PAP) less often and less regularly than national guidelines advise [2]. This translates into delayed diagnosis and inferior disease outcomes compared to heterosexual women [3]. Little data exist regarding perceptions, attitudes and behavior putting WSW at risk for cervical cancer in the Hispanic population, which comprises 3% of the WSW demographic. Hence, the objectives of this crosssectional pilot study were to (1) evaluate cervical cancer screening patterns amongst predominately Hispanic WSW, (2) investigate perceptions of health care providers and (3) assess the educational need related to this topic. We recruited our study participants from LGBTQ (Lesbian/Gay/ Bisexual/Transgender/Queer) centers or by self-referral. Respondents were self-identified WSWs, reported having a history of sex with women, were biological females and between 18 and 65 years old. We created and validated a 72-item questionnaire. Study data were collected and managed using REDCap electronic data capture tools [4]. The Institutional Review Board approval was obtained for this study (#HS-11-00467). We included 28 predominately Hispanic women (average age 37 9 years) of which 82% (80% of Hispanic participants) reported PAP screening every 1–3 years. Half of women reported no barriers to their cervical cancer screenings. For females experiencing barriers, the most common reasons were lack of insurance (21%), fear of discrimination (11%), and a lack of time to visit a doctor (11%). 75% of WSW were aware of HPV vaccinations. However, only a minority (36%) would consider getting the vaccine, and only 7% had already done so (Table 1). 37% of women perceived their personal risk of developing cervical cancer to be ‘‘very low’’ to ‘‘somewhat low’’. But over half of participants had sexual experiences with men in the past and 68% of participants reported practicing ‘‘protected sex’’, yet 39% and 75% never used condoms and dental dams, respectively. 93% of WSW believed providers should be more aware of LGBTQ specific health needs. Most women were familiar with HPV (86%), but 25% indicated they either ‘‘did not believe HPV was carcinogenic’’ or were ‘‘not sure’’. 54% did not believe ‘‘HPV could be transmitted between women’’ or were ‘‘not sure’’. Only 57% of participants were satisfied with their knowledge about HPV and PAP screening and stated their interest in educational interventions. WSW would prefer to be Table 1 Preventive gynecological care and screening history.

General practitioners in German metropolitan areas – distribution patterns and their relationship with area level measures of the socioeconomic status

BackgroundGeographical variation of the general practitioner (GP) workforce is known between rural and urban areas. However, data about the variation between and within urban areas are lacking.MethodWe analyzed distribution patterns of GP full time equivalents (FTE) in German cities with a population size of more than 500,000. We correlated their distribution with area measures of social deprivation in order to analyze preferences within neighborhood characteristics. For this purpose, we developed two area measures of deprivation: Geodemographic Index (GDI) and Cultureeconomic Index (CEI).ResultsIn total n = 9034.75 FTE were included in n = 14 cities with n = 171 districts. FTE were distributed equally on inter-city level (mean: 6.49; range: 5.12–7.20; SD: 0.51). However, on intra-city level, GP distribution was skewed (mean: 6.54; range: 1.80–43.98; SD: 3.62). Distribution patterns of FTE per 10^4 residents were significantly correlated with GDI (r = −0.49; p < 0.001) and CEI (r = −0.22; p = 0.005). Therefore, location choices of GPs were mainly positively correlated with 1) central location (r = −0.50; p < 0.001), 2) small household size of population (r = −0.50; p < 0.001) and 3) population density (r = 0.35; p < 0.001).ConclusionIntra-city distribution of GPs was skewed, which could affect the equality of access for the urban population. Furthermore, health services planners should be aware of GP location preferences. This could be helpful to better understand and plan delivery of health services. Within this process the presented Geodemographic Index (GDI) could be of use.

Improving trauma care for pregnant patients at the LAC+USC Medical Center by conducting regular multidisciplinary drills.

U p to 98 000 Americans die annually due to preventable medical errors mostly caused by deficient crisis management. Besides inadequate clinical competencies of individuals, 70% of these events can be attributed to miscommunication in health care teams. On the basis of recommendations by the Joint Commission on Accreditation of Healthcare Organizations, we identified multidisciplinary simulations (MuSs) as an effective tool to deliver better and safer health care in our facility. At the LAC + USC Medical Center, we treat pregnant trauma patients in one of the largest acute care hospitals in the United States. Rare and complex situations require teamwork for an efficient management. Our multidisciplinary response teams consist of physicians, nurses, respiratory therapists, and pharmacists who are practicing at different levels of training. Given these challenges, the setting is prone to result in suboptimal crisis management and unfavorable patient outcomes. Since 2011, we have organized regular MuSs in the emergency department. Scenarios are conducted to recreate the presentation of a pregnant patient with multiple gunshot wounds or a high-velocity car accident using a high-fidelity NOELLE Birthing Simulator. In a safe and standardized setting, participants treat the patient following a predetermined clinical pathway. This usually includes the activation of a “Code OB event” that deploys obstetrics, pediatrics, and trauma surgery teams. Observing faculty evaluates interprofessional and multidisciplinary team performance using the “Performance for Communication and Teamwork” (PACT)– long form (University of Washington). Evaluation of individual clinical knowledge and skills is based on key performance measures that are specific to each scenario and created by experts in the field. A debriefing session concludes every drill, where participants selfassess their performance and receive feedback. Traditionally, simulations are conducted to assess individual clinical competencies or team performance. Hence, the series of MuSs allowed us to identify shortcoming in these areas, for example, participants performed an inadequate surgical approach, or they did not know the timelines for delivery after circulatory breakdown of the mother. Frequently, senior team members failed to take on leadership roles, assign functions, or delegate tasks, leading to delayed patient care. On the