Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations

The HEK293 human cell lineage is widely used in cell biology and biotechnology. Here we use whole-genome resequencing of six 293 cell lines to study the dynamics of this aneuploid genome in response to the manipulations used to generate common 293 cell derivatives, such as transformation and stable clone generation (293T); suspension growth adaptation (293S); and cytotoxic lectin selection (293SG). Remarkably, we observe that copy number alteration detection could identify the genomic region that enabled cell survival under selective conditions (i.c. ricin selection). Furthermore, we present methods to detect human/vector genome breakpoints and a user-friendly visualization tool for the 293 genome data. We also establish that the genome structure composition is in steady state for most of these cell lines when standard cell culturing conditions are used. This resource enables novel and more informed studies with 293 cells, and we will distribute the sequenced cell lines to this effect

Minimally invasive classification of pediatric solid tumors using reduced representation bisulfite sequencing of cell-free DNA : a proof-of-principle study

In the clinical management of pediatric solid tumors, histological examination of tumor tissue obtained by a biopsy remains the gold standard to establish a conclusive pathological diagnosis. The DNA methylation pattern of a tumor is known to correlate with the histopathological diagnosis across cancer types and is showing promise in the diagnostic workup of tumor samples. This methylation pattern can be detected in the cell-free DNA. Here, we provide proof-of-concept of histopathologic classification of pediatric tumors using cell-free reduced representation bisulfite sequencing (cf-RRBS) from retrospectively collected plasma and cerebrospinal fluid samples. We determined the correct tumor type in 49 out of 60 (81.6%) samples starting from minute amounts (less than 10 ng) of cell-free DNA. We demonstrate that the majority of misclassifications were associated with sample quality and not with the extent of disease. Our approach has the potential to help tackle some of the remaining diagnostic challenges in pediatric oncology in a cost-effective and minimally invasive manner. Translational relevance: Obtaining a correct diagnosis in pediatric oncology can be challenging in some tumor types, especially in renal tumors or central nervous system tumors. Furthermore, the diagnostic odyssey can result in anxiety and discomfort for these children. By applying a novel technique, reduced representation bisulfite sequencing on cell-free DNA (cf-RRBS), we show the feasibility of obtaining the histopathological diagnosis with a minimally invasive test on either plasma or cerebrospinal fluid. Furthermore, we were able to derive the copy number profile or tumor subtype from the same assay. Given that primary tumor material might be difficult to obtain, in particular in critically ill children or depending on the tumor location, and might be limited in terms of quantity or quality, our assay could become complementary to the classical tissue biopsy in difficult cases

Participant evaluation of an education module on interprofessional collaboration for students in healthcare studies

BACKGROUND: Interprofessional collaboration is considered a key-factor to deliver the highest quality of care. Interprofessional collaboration (IPC) assumes a model of working together, in particular with awareness of the process of interprofessional collaboration, to develop an integrated and cohesive answer to the needs of the client/family/population. Educational modules are developed in response to a perceived need to improve interprofessional collaboration for the benefit of patientcare. Up until 2005 no explicit module on interprofessional collaboration existed in the education programs of the Antwerp University Association (AUHA). During a decade the ‘Interprofessional Collaboration In Healthcare (IPCIHC) – module’ is organised and evaluated by its participants. METHODS: One group, post-test design was used to gather data from the participating students using a structured questionnaire. Data was collected between March 2005 and March 2014 from participating final year students in healthcare educational programs. RESULTS: 3568 (84 % overall response) students evaluated the IPCIHC module from 2005 up to 2014. Over 80 % of the participants were convinced the IPCIHC increased their knowledge and changed their understanding that it will impact their future professional relationships, and felt a greater understanding about problem-solving in healthcare teams. Even though the results indicate that the goals of the IPCIHC module were achieved, less than 60 % of the participants experienced a change in attitude towards other professional groups. CONCLUSIONS: Despite the positive outcomes from the participants, the challenge still remains to keep on educating future healthcare providers in interprofessional collaboration in order to achieve an increase in interprofessional behaviour towards other professional groups. Research is needed to investigate the effectiveness of undergraduate programs on the quality and safety of patientcare in practice

Outcome indicators on interprofessional collaboration interventions for elderly

<p class="p1">  <p class="p2"><span class="s1"><strong> </strong></span><strong>Background: </strong>Geriatric care increasingly needs more multidisciplinary health care services to deliver the necessary complex and continuous care. The aim of this study is to summarize indicators of effective interprofessional outcomes for this population. <p class="p2"><strong>Method: </strong>A systematic review is performed in the Cochrane Library, Pubmed (Medline), Embase, Cinahl and Psychinfo with a search until June 2014. <p class="p2"><strong>Results: </strong>Overall, 689 references were identified of which 29 studies met the inclusion criteria. All outcome indicators were summarized in three categories: collaboration, patient level outcome and costs. Seventeen out of 24 outcome indicators within the category of ‘collaboration’ reached significant difference in advantage of the intervention group. On ‘patient outcome level’ only 15 out of 32 outcome parameters met statistical significance. In the category of ‘costs’ only one study reached statistical significance. <p class="p2"><strong>Discussion and conclusion: </strong>The overall effects of interprofessional interventions for elderly are positive, but based on heterogeneous outcomes. Outcome indicators of interprofessional collaboration for elderly with a significant effect can be summarized in three main categories: ‘collaboration’, patient level’ and ‘costs’. For ‘collaboration’ the outcome indicators are key elements of collaboration, involved disciplines, professional and patient satisfaction and quality of care. On ‘patient level’ the outcome indicators are pain, fall incidence, quality of life, independence for daily life activities, depression and agitated behaviour, transitions, length of stay in hospital, mortality and period of rehabilitation. ‘Costs’ of interprofessional interventions on short- and long-term for elderly need further investigation. When organizing interprofessional collaboration or interprofessional education these outcome indicators can be considered as important topics to be addressed. Overall more research is needed to gain insight in the process of interprofessional collaboration and so to learn to work interprofessionally

Open-cell recording of action potentials using active electrode arrays

The investigation of complex communication in cellular networks requires superior measurement tools than those available to date. Electrode arrays integrated onto silicon electronics are increasingly used to measure the electrical activity of cells in an automated and highly parallelized fashion, but they are restricted to recording extracellular potentials. Here, we report on an array of TiN electrodes built using standard silicon electronics for intracellular action potential recording. Intracellular access, possible at each of the 16 384 electrodes on the chip, was accomplished by local membrane electroporation using electrical stimulation with subcellular, micrometer-sized electrodes. Access to the cell interior was transient and could be tuned in duration by adapting the electroporation protocol. Intracellular sensing was found to be minimally invasive in the short and long-term, allowing consecutive intracellular recordings from the same cell over the course of days. Finally, we applied this method to investigate the effect of an ion channel blocker on cardiac electrical activity. This technique opens the door to massively parallel, long-term intracellular recording for fundamental electrophysiology and drug screening.status: publishe

Single-cell recording and stimulation with a 16k micro-nail electrode array integrated on a 0.18 μm CMOS chip

To cope with the growing needs in research towards the understanding of cellular function and network dynamics, advanced micro-electrode arrays (MEAs) based on integrated complementary metal oxide semiconductor (CMOS) circuits have been increasingly reported. Although such arrays contain a large number of sensors for recording and/or stimulation, the size of the electrodes on these chips are often larger than a typical mammalian cell. Therefore, true single-cell recording and stimulation remains challenging. Single-cell resolution can be obtained by decreasing the size of the electrodes, which inherently increases the characteristic impedance and noise. Here, we present an array of 16 384 active sensors monolithically integrated on chip, realized in 0.18 μm CMOS technology for recording and stimulation of individual cells. Successful recording of electrical activity of cardiac cells with the chip, validated with intracellular whole-cell patch clamp recordings are presented, illustrating single-cell readout capability. Further, by applying a single-electrode stimulation protocol, we could pace individual cardiac cells, demonstrating single-cell addressability. This novel electrode array could help pave the way towards solving complex interactions of mammalian cellular networks.status: publishe