Discovery of dachshund 2 protein as a novel biomarker of poor prognosis in epithelial ovarian cancer

<p>Abstract</p> <p>Background</p> <p>The Dachshund homolog 2 (<it>DACH2</it>) gene has been implicated in development of the female genital tract in mouse models and premature ovarian failure syndrome, but to date, its expression in human normal and cancerous tissue remains unexplored. Using the Human Protein Atlas as a tool for cancer biomarker discovery, DACH2 protein was found to be differentially expressed in epithelial ovarian cancer (EOC). Here, the expression and prognostic significance of DACH2 was further evaluated in ovarian cancer cell lines and human EOC samples.</p> <p>Methods</p> <p>Immunohistochemical expression of DACH2 was examined in tissue microarrays with 143 incident EOC cases from two prospective, population-based cohorts, including a subset of benign-appearing fallopian tubes (n = 32). A nuclear score (NS), i.e. multiplier of staining fraction and intensity, was calculated. For survival analyses, cases were dichotomized into low (NS < = 3) and high (NS > 3) using classification and regression tree analysis. Kaplan Meier analysis and Cox proportional hazards modelling were used to assess the impact of DACH2 expression on survival. DACH2 expression was analysed in the cisplatin sensitive ovarian cancer cell line A2780 and its cisplatin resistant derivative A2780-Cp70. The specificity of the DACH2 antibody was tested using siRNA-mediated silencing of DACH2 in A2780-Cp70 cells.</p> <p>Results</p> <p>DACH2 expression was considerably higher in the cisplatin resistant A2780-Cp70 cells compared to the cisplatin-sensitive A2780 cells. While present in all sampled fallopian tubes, DACH2 expression ranged from negative to strong in EOC. In EOC, DACH2 expression correlated with several proteins involved in DNA integrity and repair, and proliferation. DACH2 expression was significantly higher in carcinoma of the serous subtype compared to non-serous carcinoma. In the full cohort, high DACH2 expression was significantly associated with poor prognosis in univariable analysis, and in carcinoma of the serous subtype, DACH2 remained an independent factor of poor prognosis.</p> <p>Conclusions</p> <p>This study provides a first demonstration of DACH2 protein being expressed in human fallopian tubes and EOC, with the highest expression in serous carcinoma where DACH2 was found to be an independent biomarker of poor prognosis. Future research should expand on the role of DACH2 in ovarian carcinogenesis and chemotherapy resistance.</p

Intrarater Reliability of Digital Thermography in Detecting Pin Site Infection:A Proof of Concept Study

AIM AND OBJECTIVE: The purpose of this study was to explore the capability and Intrarater reliability of thermography in detecting pin site infection. MATERIALS AND METHODS: This is an explorative proof of concept study. Clinical assessment of pin sites was performed by one examiner with the Modified Gordon Pin Infection Classification from grade 0 to 6. Thermography of the pin sites was performed with a FLIR C3 camera. The analysis of the thermographic images was done in the software FLIR Tools. The maximum skin temperature around the pin site and the maximum temperature for the whole thermographic picture were measured. An Intrarater agreement was established and test-retests were performed with different camera angles. RESULTS: Thirteen (four females, nine males) patients (age 9–72 years) were included. Indications for frames: Fracture (n=4), two deformity correction, one lengthening and six bone transport. Days from surgery to thermography ranged from 27 to 385 days. Overall, 231 pin sites were included. Eleven pin sites were diagnosed with early signs of infection: five grade 1, five grade 2 and one grade 3. Mean pin site temperature for each patient was calculated, varied between patients from 29.0°C to 35.4°C (mean 33.9°C). With 34°C as cut-off value for infection, sensitivity was 73%; specificity, 67%; positive predictive value, 10%; and negative predictive value, 98%. Intrarater agreement for thermography was ICC 0.85 (0.77–0.92). The temperature measured was influenced by the camera positioning in relation to the pin site with a variance of 0.2. CONCLUSIONS: Measurements of pin site temperature using the hand-held FLIR C3 infrared camera was a reliable method and the temperature was related to infection grading. CLINICAL SIGNIFICANCE: This study demonstrated that digital thermography with a hand-held camera might be used for monitoring the pin sites after operations to detect early infection. HOW TO CITE THIS ARTICLE: Rahbek O, Husum HC, Fridberg M, et al. Intrarater Reliability of Digital Thermography in Detecting Pin Site Infection: A Proof of Concept Study. Strategies Trauma Limb Reconstr 2021;16(1):1–7

Broadening the foundation for the study of childhood connectedness to nature

The spatial aspect of access to nature experience is considered a key factor for studying school-age educare and connectedness to nature. While the standard approach for questions of connectedness to nature is to study at the individual level using methods such as observations, psychometric scaling, and interviews, less common are spatial methods applied to structural or collective aspects of these questions; connectedness to nature study rarely considers the human relationship with nature across sociocultural/structural/institutional levels. Spatial analysis is presented as a step toward a broader consideration of connectedness to nature; careful consideration of connectedness to nature/disconnection must explore the forces beyond the individual shaping access and opportunity. Specifically, the study considers access through proximity to nature from school-age educare sites in the Swedish city of Malmö. Using spatial methodology, proximity to nature was measured at 67 school-aged educare sites. The results provide a complex picture of a range from high to low-quality access to nature for children at the sites. The results help highlight the importance of access via proximity while also opening the door to a mix of other sociocultural/structural/institutional factors to be considered in support of children’s access to nature experience

Preparing Infection Detection Technology for Hospital at Home after Lower Limb External Fixation

BACKGROUND: Patients with severe bone fractures and complex bone deformities are treated by orthopedic surgeons with external fixation for several months. During this long treatment period, there is a high risk of inflammation and infection at the superficial skin area (pin site). This can develop into a devastating, sometimes fatal, and always costly condition of deep bone infection. OBJECTIVE: For pin site infection surveillance, thermography technology could be the solution to build an objective and continuous home-based remote monitoring tool to avoid frequent nursing care and hospital visits. However, future studies of infection monitoring require a preliminary step to automate the process of locating and detecting the pin sites in thermal images reliably for temperature measurement, and this step is the aim of this study. METHODS: This study presents an automatic approach for identifying and annotating pin sites on visible images using bounding boxes and transferring them to the corresponding thermal images for temperature measurement. The pin site is detected by applying deep learning-based object detection architecture YOLOv5 with a novel loss evaluation and regression method, control distance intersection over union. Furthermore, we address detecting pin sites in a practical environment (home setting) accurately through transfer learning. RESULTS AND CONCLUSION: The proposed model offers the pin site detection in 1.8 ms with a high precision of 0.98 and enables temperature information extraction. Our work for automatic pin site annotation on thermography paves the way for future research on infection assessment on thermography

Validation of podocalyxin-like protein as a biomarker of poor prognosis in colorectal cancer

Background: Podocalyxin-like 1 (PODXL) is a cell-adhesion glycoprotein and stem cell marker that has been associated with an aggressive tumour phenotype and adverse outcome in several cancer types. We recently demonstrated that overexpression of PODXL is an independent factor of poor prognosis in colorectal cancer (CRC). The aim of this study was to validate these results in two additional independent patient cohorts and to examine the correlation between PODXL mRNA and protein levels in a subset of tumours. Method: PODXL protein expression was analyzed by immunohistochemistry in tissue microarrays with tumour samples from a consecutive, retrospective cohort of 270 CRC patients (cohort 1) and a prospective cohort of 337 CRC patients (cohort 2). The expression of PODXL mRNA was measured by real-time quantitative PCR in a subgroup of 62 patients from cohort 2. Spearman's Rho and Chi-Square tests were used for analysis of correlations between PODXL expression and clinicopathological parameters. Kaplan Meier analysis and Cox proportional hazards modelling were applied to assess the relationship between PODXL expression and time to recurrence (TTR), disease free survival (DFS) and overall survival (OS). Results: High PODXL protein expression was significantly associated with unfavourable clinicopathological characteristics in both cohorts. In cohort 1, high PODXL expression was associated with a significantly shorter 5-year OS in both univariable (HR = 2.28; 95% CI 1.43-3.63, p = 0.001) and multivariable analysis (HR = 2.07; 95% CI 1.25-3.43, p = 0.005). In cohort 2, high PODXL expression was associated with a shorter TTR (HR = 2.93; 95% CI 1.26-6.82, p = 0.013) and DFS (HR = 2.44; 95% CI 1.32-4.54, p = 0.005), remaining significant in multivariable analysis, HR = 2.50; 95% CI 1.05-5.96, p = 0.038 for TTR and HR = 2.11; 95% CI 1.13-3.94, p = 0.019 for DFS. No significant correlation could be found between mRNA levels and protein expression of PODXL and there was no association between mRNA levels and clinicopathological parameters or survival. Conclusions: Here, we have validated the previously demonstrated association between immunohistochemical expression of PODXL and poor prognosis in CRC in two additional independent patient cohorts. The results further underline the potential utility of PODXL as a biomarker for more precise prognostication and treatment stratification of CRC patients

Protein kinases and phosphatases in B-cell lymphoma

Around 2000 persons are diagnosed with lymphoma in Sweden each year. There are many subgroups described for this form of cancer and the great majority is derived from B-cells. The most common subgroup is Diffuse large B-cell lymphoma (DLBCL), a highly aggressive disease where only half of the patients are cured. The lymphoma types seen in adults and children vary and the dominant form in children is the B-cell derived Burkitt lymphoma (BL). In contrast to DLBCL, high survival rates in BL are obtained but more research on normal- and tumor B-cells is needed for a better understanding of the cellular mechanisms underlying lymphoma pathogenesis. Phosphorylation and dephosphorylation of enzymes are key events in signal transduction in cells. Protein kinases phosphorylate enzymes while protein phosphatases dephosphorylate them, and these opposite actions must be carefully balanced for normal cell division to occur. The aim of this study was to characterize B-cell lymphomas with emphasis on a panel of kinases and phosphatases previously described as oncogenes (kinases) and tumorsuppressors (phosphatases) in human cancers. We analyzed DLBCL tissue and found the kinases PKC-β II and ZAP70 to be stronger expressed at protein level in a subgroup of the disease associated with poor survival. We further report enhanced activation of DLBCL cells through the B-cell receptor when ZAP70 is introduced. Also, the protein expression of the phosphatases HePTP and PTEN are weaker in BL tissue from children compared to in non-malignant pediatric controls. Our results indicate important roles for the described kinases and phosphatases in B-cell lymphomas

Teachers’ and children’s use of words during early childhood STEM teaching supported by robotics

With science and digitalisation emphasised further in the new Swedish preschool curriculum, there is a need to clarify teachers’ role in educating children in and about these areas. With research pointing out the importance of a conscious language use in STEM teaching, we here focus on words used by teachers and children during inquiry-based STEM activities in five different preschools. Bers’ powerful ideas about early childhood computational thinking (Bers 2018. Coding as a Playground. New York: Routledge.) were used for analysis and results highlight how digital programming and use of robots can promote a more versatile use of robotic words, compared to analogue, ‘unplugged’, programming without robots. Furthermore, it is also found that use of precise decontextualised language by the teacher seems to stimulate children’s use of words related to STEM and the object of learning. The findings add to the discussion about how teachers can scaffold children’s learning by inquiry teaching of STEM supported by robotics