Enforcing differentiation of cancer stem cells is considered as a potential strategy to sensitize colorectal cancer cells to irradiation and chemotherapy. Activation of the unfolded protein response, ...due to endoplasmic reticulum (ER) stress, causes rapid stem cell differentiation in normal intestinal and colon cancer cells. We previously found that stem cell differentiation was mediated by a Protein kinase R-like ER kinase (PERK) dependent arrest of mRNA translation, resulting in rapid protein depletion of WNT-dependent transcription factor c-MYC. We hypothesize that ER stress dependent stem cell differentiation may rely on the depletion of additional transcriptional regulators with a short protein half-life that are rapidly depleted due to a PERK-dependent translational pause. Using a novel screening method, we identify novel transcription factors that regulate the intestinal stem cell fate upon ER stress. ER stress was induced in LS174T cells with thapsigargin or subtilase cytotoxin (SubAB) and immediate alterations in nuclear transcription factor activity were assessed by the CatTFRE assay in which transcription factors present in nuclear lysate are bound to plasmid DNA, co-extracted and quantified using mass-spectrometry. The role of altered activity of transcription factor CtBP2 was further examined by modification of its expression levels using CAG-rtTA3-CtBP2 overexpression in small intestinal organoids, shCtBP2 knockdown in LS174T cells, and familial adenomatous polyposis patient-derived organoids. CtBP2 overexpression organoids were challenged by ER stress and ionizing irradiation. We identified a unique set of transcription factors with altered activation upon ER stress. Gene ontology analysis showed that transcription factors with diminished binding were involved in cellular differentiation processes. ER stress decreased CtBP2 protein expression in mouse small intestine. ER stress induced loss of CtBP2 expression which was rescued by inhibition of PERK signaling. CtBP2 was overexpressed in mouse and human colorectal adenomas. Inducible CtBP2 overexpression in organoids conferred higher clonogenic potential, resilience to irradiation-induced damage and a partial rescue of ER stress-induced loss of stemness. Using an unbiased proteomics approach, we identified a unique set of transcription factors for which DNA-binding activity is lost directly upon ER stress. We continued investigating the function of co-regulator CtBP2, and show that CtBP2 mediates ER stress-induced loss of stemness which supports the intestinal stem cell state in homeostatic stem cells and colorectal cancer cells.
Postoperative rehabilitation after primary total hip arthroplasty (p-THA) differs between the Netherlands and Germany. Aim is to compare clinical effectiveness and to get a first impression of cost ...effectiveness of Dutch versus German usual care after p-THA.
A transnational prospective controlled observational trial. Clinical effectiveness was assessed with self-reported questionnaires and functional tests. Measurements were taken preoperatively and 4 weeks, 12 weeks, and 6 months postoperatively. For cost effectiveness, long-term economic aspects were assessed from a societal perspective.
124 working-age patients finished the measurements. German usual care leads to a significantly larger proportion (65.6% versus 47.5%) of satisfied patients 12 weeks postoperatively and significantly better self-reported function and Five Times Sit-to-Stand Test (FTSST) results. German usual care is generally 45% more expensive than Dutch usual care, and 20% more expensive for working-age patients. A scenario analysis assumed that German patients work the same number of hours as the Dutch, and that productivity costs are the same. This analysis revealed German care is still more expensive but the difference decreased to 8%.
German rehabilitation is clinically advantageous yet more expensive, although comparisons are less straightforward as the socioeconomic context differs between the two countries.
The study is registered in the German Registry of Clinical Trials (DRKS00011345, 18/11/2016).
During the suckling‐to‐weaning transition, the intestinal epithelium matures, allowing digestion of solid food. Transplantation experiments with rodent fetal epithelium into subcutaneous tissue of ...adult animals suggest that this transition is intrinsically programmed and occurs in the absence of dietary or hormonal signals. Here, we show that organoids derived from mouse primary fetal intestinal epithelial cells express markers of late fetal and neonatal development. In a stable culture medium, these fetal epithelium‐derived organoids lose all markers of neonatal epithelium and start expressing hallmarks of adult epithelium in a time frame that mirrors epithelial maturation in vivo. In vitro postnatal development of the fetal‐derived organoids accelerates by dexamethasone, a drug used to accelerate intestinal maturation in vivo. Together, our data show that organoids derived from fetal epithelium undergo suckling‐to‐weaning transition, that the speed of maturation can be modulated, and that fetal organoids can be used to model the molecular mechanisms of postnatal epithelial maturation.
Synopsis
Organoids derived from mouse primary fetal intestinal epithelial cells undergo suckling‐to‐weaning transition independent of stromal cells and can be used to model the molecular mechanisms of postnatal epithelial development.
Mouse fetal intestinal organoids undergo the suckling‐to‐weaning transition in vitro.
Fetal intestinal organoids express functional adult brush border enzymes over time.
Dexamethasone can accelerate intestinal epithelial maturation in vitro.
Fetal intestinal organoids can be used to model postnatal epithelial development.
Organoids derived from mouse primary fetal intestinal epithelial cells undergo suckling‐to‐weaning transition independent of stromal cells and can be used to model the molecular mechanisms of postnatal epithelial development.
The High‐Dynamic Double‐Crystal Monochromator (HD‐DCM) is a mechatronic system with unique control‐based architecture and deep paradigm changes as compared with traditional beamline monochromators. ...Aiming at unprecedented inter‐crystal positioning stability in vertical‐bounce double‐crystal monochromators (DCMs) of the order of 10 nrad RMS (1 Hz to 2.5 kHz), and not only in fixed‐energy but also in fly‐scan operation, it has been developed according to a `first‐time right' predictive design approach for hard X‐ray beamlines at Sirius, the fourth‐generation light source at the Brazilian Synchrotron Light Laboratory (LNLS/CNPEM). This work explores some of the challenges that emerge with this new technology and presents the latest commissioning results that demonstrate the unparallel performances of the HD‐DCM at the undulator‐based EMA (Extreme Methods of Analysis) beamline at Sirius. With the enabled fast spectroscopy fly‐scan possibilities, a new energy‐tuning evaluation method, based on wave‐propagation simulations, becomes part of a motion‐oriented analysis that is carried out to derive the multi‐axis non‐linear positioning problem, covering not only energy selection and fixed exit in the HD‐DCM but also the emission spectrum of an adjustable‐phase undulator (APU). The HD‐DCM control scheme and its flexible operation modes are described in detail as well. Furthermore, a new integration topology between the HD‐DCM and EMA's APU, coming already close to ultimate motion levels, is described and validated.
The High‐Dynamic Double‐Crystal Monochromator that has been developed for the fourth‐generation light source Sirius/LNLS is explored, with its unprecedented inter‐crystal stability performance around 10 nrad RMS both at fixed‐energy and continuous fly‐scan. Given the new paradigm and the unique enabling technology of its high‐performance mechatronic system, the engineering and operational aspects for the next‐level beamline experimental possibilities with a double‐crystal monochromator are comprehensively discussed. Fly‐scan spectroscopy scientific commissioning results are also presented.
ICES: Data, Discovery, Better Health Schull, Michael J; Azimaee, Mahmoud; Marra, Marcel ...
International journal of population data science,
03/2020, Letnik:
4, Številka:
2
Journal Article
Recenzirano
Odprti dostop
ICES was founded in 1992 to study the health care system and promote effective, efficient and equitable health care. Over 27 years later, the goal remains largely unchanged, though the institute has ...grown in size and impact. Created as an independent not-for-profit research institute and given what was, at the time, unprecedented access to administrative health data records for the population of Ontario, ICES’ initial focus was to better understand the delivery of hospital services and translate its findings into better health care and policy. From modest beginnings with a handful of researchers located in a few hospital offices, ICES has grown to encompass a community of almost 500 scientists and staff across a network of seven physical sites in Ontario. The original focus on hospital-based services has expanded significantly and now includes research and analysis of community-based health services, health policy, Indigenous health, social determinants of health, and data science.
An extensive protein–protein interaction network has been identified between proteins implicated in inherited ataxias. The protein sacsin, which is mutated in the early-onset neurodegenerative ...disease autosomal recessive spastic ataxia of Charlevoix-Saguenay, is a node in this interactome. Here, we have established the neuronal expression of sacsin and functionally characterized domains of the 4579 amino acid protein. Sacsin is most highly expressed in large neurons, particularly within brain motor systems, including cerebellar Purkinje cells. Its subcellular localization in SH-SY5Y neuroblastoma cells was predominantly cytoplasmic with a mitochondrial component. We identified a putative ubiquitin-like (UbL) domain at the N-terminus of sacsin and demonstrated an interaction with the proteasome. Furthermore, sacsin contains a predicted J-domain, the defining feature of DnaJ/Hsp40 proteins. Using a bacterial complementation assay, the sacsin J-domain was demonstrated to be functional. The presence of both UbL and J-domains in sacsin suggests that it may integrate the ubiquitin–proteasome system and Hsp70 function to a specific cellular role. The Hsp70 chaperone machinery is an important component of the cellular response towards aggregation prone mutant proteins that are associated with neurodegenerative diseases. We therefore investigated the effects of siRNA-mediated sacsin knockdown on polyglutamine-expanded ataxin-1. Importantly, SACS siRNA did not affect cell viability with GFP-ataxin-130Q, but enhanced the toxicity of GFP-ataxin-182Q, suggesting that sacsin is protective against mutant ataxin-1. Thus, sacsin is an ataxia protein and a regulator of the Hsp70 chaperone machinery that is implicated in the processing of other ataxia-linked proteins.
The assessment of health-related quality of life (HRQoL) is important for health outcomes research, disease modeling studies and comparisons of different healthcare interventions. Yet, only a few ...tools are available to assess HRQoL in 0-1-year-old infants. Furthermore, there is a need for an instrument able to assess HRQoL with a single, standardized, overall score in the first year of life. Here we described the development of the Infant health-related Quality of life Instrument (IQI), a generic, preference-based instrument that can be administered through a mobile application for assessing HRQoL in 0-1-year-old infants.
A multi-step development process began by extracting candidate health concepts from relevant measures identified by two literature searches. Next, three panels, with experts from Asia, Europe, New Zealand and United States of America, and two surveys, with primary caregivers in New Zealand, Singapore, and the United Kingdom, evaluated the relevance of the candidate health concepts, organized them into attributes based on their similarities, explored alternative attributes and generated response scales. Additional interviews assessed the cross-cultural interpretability, parents' understanding of health attributes, and the usability of the mobile application.
The final list of 7 health attributes included in the IQI consisted of sleeping, feeding, breathing, stooling/poo, mood, skin, and interaction. The users' experiences with the mobile application were generally positive.
The IQI is the first generic, preference-based, instrument designed to assess overall HRQoL in 0-1-year old infants. It is short and easy-to-administer through a mobile application. Moreover, close attention was paid to the opinions of the infants' primary caregivers during the instrument and mobile application development process.
Summary Introduction Home mechanical ventilation (HMV) in the Netherlands is normally initiated in hospital, but this is expensive and often a burden for the patient. In this randomised controlled ...study we investigated whether initiation of HMV at home in patients with chronic respiratory failure is non-inferior to an in hospital based setting. Methods Seventy-seven patients were included, of which 38 patients started HMV at home. All patients were diagnosed with chronic respiratory failure due to a neuromuscular or thoracic cage disease. Primary outcome was the arterial carbon dioxide ( P aCO2 ) while quality of life and costs were secondary outcomes. Telemonitoring was used in the home group to provide therapeutic information, for example; transcutaneous carbon dioxide, oxygen saturation and ventilator information, to the caregivers. Follow-up was six months. Results P aCO2 , improved by 0.72 (SE ± 0.16) kPa in the hospital group and by 0.91 (±0.20) in the home group, both improvements being significant and the latter clearly not inferior. There were also significant improvements in quality of life in both groups, again not being inferior with home treatment. Conclusion This study is the first to show that initiation of HMV at home in a selective group of patients with chronic respiratory failure is as effective for gas exchange and quality of life as hospital initiation. In addition we found that it is safe, technically feasible and that more than € 3000 per patient can be saved compared to our standard care.
Biodegradable fixation systems could reduce/delete the problems associated with titanium plate removal. This means less surgical discomfort, and a reduction in costs.
The aim of the present study was ...to compare the cost-effectiveness between a biodegradable and a titanium system in Maxillofacial surgery.
This multicenter RCT was performed in the Netherlands from December 2006 to July 2009. Included were 230 patients who underwent a bilateral sagittal split osteotomy (BSSO), a Le Fort-I osteotomy, or a bi-maxillary osteotomy and those treated for fractures of the mandible, maxilla, or zygoma. The patients were randomly assigned to a titanium group (KLS Martin) or to a biodegradable group (Inion CPS). Costs were assessed from a societal perspective. Health outcomes in the incremental cost-effectiveness ratio (ICER) were bone healing (8 weeks) and plate removal (2 years).
In 25 out of the 117 patients who were randomized to the biodegradable group, the maxillofacial surgeon made the decision to switch to the titanium system intra-operatively. This resulted in an Intention-To-Treat (ITT-)analysis and a Treatment-Received (TR-) analysis. Both analyses indicated that operations performed with titanium plates and screws had better health outcomes. In the TR-analysis the costs were lower in the biodegradable group, in the ITT-analysis costs were lower in the titanium group.
The difference in costs between the ITT and the TR analyses can be explained by the intra-operative switches: In the TR-analysis the switches were analysed in the titanium group. In the ITT-analysis they were analysed in the biodegradable group. Considering the cost-effectiveness the titanium system is preferable to the biodegradable system in the regular treatment spectrum of mandibular, Le Fort-I, and zygomatic fractures, and BSSO's, Le Fort-I osteotomies and bimaxillary osteotomies.
Controlled-Trials.com ISRCTN 44212338.
PDF
