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139 results for “Load cell”
Data description: Deprivation of loading during early healing of rat Achilles tendons affects extracellular matrix composition and structure, and reduces cell density and cell alignment
<p><a name="_Hlk158643946"></a><strong>Data description: Deprivation of loading during early healing of rat Achilles tendons affects extracellular matrix composition and structure, and reduces cell density and cell alignment</strong></p> <p><em>Malin Hammerman, Maria Pierantoni, Hanna Isaksson<sup> *</sup>, Pernilla Eliasson <sup>*</sup></em></p> <p><em><sup>* </sup></em><em>joint<sup> </sup>last authors</em></p> <p>This dataset contains microscope images obtained from sections of healing and intact rat Achilles tendons undergoing different in vivo loading protocols and different time points post-transection. The data presented are the full resolution microscope images available in lower resolution in the accompanying manuscript’s Supplementary Figures 4-6.</p> <p>Each zipped folders contain images (tif-files) from all time-points for each respective staining and loading group. </p> <ul> <li>Col1: Sections stained with Collagen 1 antibodies</li> <li>Col3: Sections stained with Collagen 3 antibodies</li> <li>Elastin: Sections stained with Elastin antibodies</li> <li>Full_loading: Free cage activity</li> <li>Reduced_loading: Paralysis of the calf muscle with Botox</li> <li>Minimal_loading: Botox combined with joint fixation using a steel-orthosis</li> <li>Intact_reference: Contralateral uninjured Achilles tendons, used as reference</li> </ul> <p>More description of the datasets inside the zipped files are available below and in the file 'Data Description.pdf'</p> <p> </p> <p><strong>Brief re-cap of methods</strong></p> <p>Histological analysis was performed on healing Achilles tendons from Female Sprague-Dawley rats, specific-pathogen free (11-12 weeks, weight 299 ± 15 g), that had undergone full transection [13] of the right Achilles tendon, and been exposed to different levels of loading. Altered loading was imposed through two mechanisms. Reduced loading involved intramuscular Botox injections in the right calf muscles to induce plantar flexor muscle paralysis [24]. Additionally, the rats in the minimal loading group received a steel-orthosis around their right hindlimb directly after surgery [24].</p> <p>Snap frozen tendons in OCT were sectioned longitudinally (7 μm thickness) and stained with immunofluorescent staining for collagen 1, collagen 3, or elastin. Sections were counterstained with DAPI followed by mounting. The tissue sections were imaged under a microscope (DMi8, Leica Microsystems, Wetzlar, Germany, with a Hamamatsu Orca LT Flash sCMOS camera) where fluorescence was detected at 550 nm (secondary antibody Alexa Fluor 594), 470 nm (secondary antibody Alexa Fluor 488) and 385 nm (DAPI), and exposure time was held constant for each color channel regarding magnification and staining.</p> <p>Mapping images of the entire tendon were obtained for one section per group (n=1 per healing time, loading group and ECM matrix protein). All images were adjusted to the negative control, where the primary antibody was omitted, to correct for unspecific antibody detection.</p> <p><strong>Microscope images and description of file-names </strong></p> <p>All data is presented in the form of .tif files. Please refer to the scale bars in the images. All image-files are named using the following abbreviations, as described below. As an example, the file name “Tendon_col1_FL_1W_col1.tif” refers to a tendon section stained for collagen 1 from a rat exposed to full loading for a period of 1 week after tendon transection, where only the channel for collagen 1 is shown, whereas “Tendon_col1_FL_1W_merged.tif” includes the channels for both staining for collagen 1 and DAPI of the same section.</p> <p>Col1: Sections stained with Collagen 1 antibodies<br>Col3: Sections stained with Collagen 3 antibodies<br>Elastin: Sections stained with Elastin antibodies<br>dapi: Sections stained with 4',6-Diamidino-2-Phenylindole Dihydrochloride.<br>FL: Full loading (free cage activity),<br>RL: Reduced loading (paralysis of the calf muscle with Botox),<br>ML: Minimal loading (Botox combined with joint fixation using a steel-orthosis)<br>IT: Intact contralateral Achilles tendons, used as reference.</p> <p>1W: Healing time point 1 week after transection<br>2W: Healing time point 2 weeks after transection<br>3W: Healing time point 3 weeks after transection<br>20W: Healing time point 20 weeks after transection</p> <p><strong>Settings for brightness and contrast</strong></p> <p><em>Collagen 1</em><br>1w FL 2000-12 000, UL 4000-10 000, ML 4000-12 000<br>2w FL 2500-10 000, UL 4000-10 000, ML 5000-12 000<br>3w FL 2000-12 000, UL 3500-13 000, ML 3500-14 000<br>12w FL 3000-12 000<br>20w FL 3000-11 000<br>IT 2000-8 000</p> <p>Collagen 3<br>1w FL 3000-12 000, UL 4000-10 000, ML 4000-13 000<br>2w FL 2000 - 7 000, UL 2500-12 000, ML 2000-12 000<br>3w FL 2000-12 000, UL 3500-13 000, ML 3500-14 000<br>12w FL 3000-12 000<br>20w FL 2000-12 000<br>IT 3000-12 000</p> <p>Elastin<br>1w FL 4000-10 000, UL 5000 - 8000, ML 3500-12 000<br>2w FL 3000-12 000, UL 3000-12 000, ML 3000-12 000<br>3w FL 2500-12 000, UL 2000-12 000, ML 2500-12 000,<br>12w FL 3500-12 000<br>20w FL 3500-12 000<br>IT 2000-12 000</p>
Dataset for the paper "High Loading of Single Atomic Iron Sites in Pyrolysed Fe-NC Oxygen Reduction Catalysts for Proton Exchange Membrane Fuel Cells", DOI:10.1038/s41929-022-00772-9
<p>The data in this spreadsheet was used to produce the figures in the paper </p> <p>Authors: Asad Mehmood, Mengjun Gong, Frédéric Jaouen, Aaron Roy, Andrea Zitolo, Anastassiya Khan, Moulay-Tahar Sougrati, Mathias Primbs, Alex Martinez Bonastre, Dash Fongalland, Goran Drazic, Peter Strasser, Anthony Kucernak</p> <p>Title: High Loading of Single Atomic Iron Sites in Pyrolysed Fe-NC Oxygen Reduction Catalysts for Proton Exchange Membrane Fuel Cells</p> <p>Journal: Nature Materials</p> <p>DOI: 10.1038/s41929-022-00772-9</p> <p>Please cite the above reference if you wish to use this data </p> <p> </p> <p>DOI of data: 10.5281/zenodo.6411262</p>
CXCL12-loaded-hydrogel (CLG): a new device for metastatic Circulating Tumor Cells (CTCs) capturing and characterization
<p><strong>Background</strong>: Circulating Tumor Cells (CTCs) represent a small, heterogeneous population that comprise the minority of cells able to develop metastasis. To trap and characterize CTCs with metastatic attitude, a CXCL12-loaded hyaluronic-gel (CLG) was developed. CXCR4+cells with invasive capability would infiltrate CLG.<br><strong>Methods</strong>: Human colon, renal, lung and ovarian cancer cells (HT29, A498, H460 and OVCAR8 respectively) were seeded on 150 µl Empty Gels (EG) or 300 ng/ml CXCL12 loaded gel (CLG) and allowed to infiltrate for 16 hours. Gels were then digested and fixed with 2% FA-HAse for human cancer cell enumeration or digested with HAse and cancer cells recovered. CLG-recovered cells migrated toward CXCL12 and were tested for colonies/spheres formation. Moreover, CXCR4, E-Cadherin and Vimentin expression was assessed through flow cytometry and RT-PCR. The clinical trial “TRAP4MET” recruited 48 metastatic/advanced cancer patients (8 OC, 8 LC, 8 GBM, 8 EC, 8 RCC and 8 EC). 10 cc whole blood were devoted to PBMCs extraction (7cc) and ScreenCell™ filters (3cc) CTCs evaluation. Ficoll-isolated patient’s PBMCs were seeded over CLG and allowed to infiltrate for 16 hours; gels were digested and fixed with 2% FA-HAse, cells stained and DAPI+/CD45-/pan-CK+ cells enumerated as CTCs.<br><strong>Results</strong>: Human cancer cells infiltrate CLG more efficiently than EG (CLG/EG ratio 1.25 for HT29/ 1.58 for A498/ 1.71 for H460 and 2.83 for OVCAR8). CLG- recovered HT29 cells display hybrid-mesenchymal features [low E-cadherin (40%) and high vimentin (235%) as compared to HT29], CXCR4 two-fold higher than HT29, efficiently migrate toward CXCL12 (two-fold higher than HT29) and developed higher number of colonies (171±21 for HT29-CLG vs 131±8 colonies for HT29) /larger spheres (spheroid area: 26561±6142 µm2 for HT29-CLG vs 20297±7238 for HT29). In TRAP4MET clinical trial, CLG-CTCs were isolated in 8/8 patients with OC, 6/8 with LC, 6/8 with CRC, 8/8 with EC, 8/8 with RCC cancer and 5/8 with GBM. Interestingly, in OC, LC and GBM, CLG isolated higher number of CTCs as compared to the conventional ScreenCell™ (CLG/SC ratio=1.88 for OC, 2.47 for LC and 11.89 for GBM). Bland and Altman blot analysis and Passing and Bablok regression analysis showed concordance between the methodological approaches but indicate that SC and CLG are not superimposable suggesting that the two systems select cells with different features.<br><strong>Conclusion</strong>: CLG might represent a new and easy tool to isolate invasive CTCs in multiple cancers such as OC, LC and GBM at today orphan of reliable methods to consistently detect CTCs. </p>
Dynamic interactome of the MHC I peptide loading complex in human dendritic cells - Source II
<p>Source data underlying MS data set (SFig.2). Datafile comprises MS raw files + MaxQuant output files.</p>
Dynamic interactome of the MHC I peptide loading complex in human dendritic cells - Source III
<p>Source data underlying Raji cell data set (SFig.3). Datafile comprises MS raw data + MaxQuant output files.</p>
Insulin Sensitivity and Beta Cell Function After Carbohydrate Loading in Hip Replacement Surgery
ClinicalTrials.gov study NCT01774084. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Long-term performance of PEM water electrolysis cells with 3D printed electrodes and low catalyst loading
<p>Original data for the article with the same title</p>
Dataset for the paper "Nanostructured Catalyst Layer Allowing Production of Ultralow Loading Electrodes for Polymer Electrolyte Membrane Fuel Cells with Superior Performance" published in ACS Appl. Energy Mater.
<p>The data in this spreadsheet was used to produce the figures in the paper</p><p>Authors:</p><p>Colleen Jackson, Michalis Metaxas, Jack Dawson, Anthony Kucernak</p><p>Title:</p><p>Nanostructured Catalyst Layer Allowing Production of Ultralow Loading Electrodes for Polymer Electrolyte Membrane Fuel Cells with Superior Performance</p><p>Journal:</p><p>ACS Appl. Energy Mater. </p><p>DOI:</p><p>10.1021/acsaem.3c01987</p><p>Please cite the above reference if you wish to use this data</p><p>DOI of data:</p><p>10.5281/zenodo.10256698</p>
A method to investigate Pathological Variant load of Mitochondrial DNA in single human Mesenchyma stem cells
<p>Mesenchymal stem cells (MSCs) are the parent cells to many cells of the musculoskeletal system including osteoblasts. Previous work has shown that in mouse models mitochondrial DNA (mtDNA) pathological variants lead to dysfunction of the respiratory chain of these osteoblasts and the premature development of age related osteoporosis. An increased rate of respiratory chain deficiency has also been observed in human osteoblasts. mtDNA mutates at an enhanced rate compared to nuclear DNA and has been linked to the ageing process in humans. </p> <p>We have developed and optimised an experimental pipeline to isolate single MSCs using flow cytometry before performing next generation sequencing to analyse the pathogenic variant load of 13 patients aged 22-88 to assess if pathogenic variants exist in mesenchymal stem cells and if there are age related changes.</p> <p>Across all patients and all cells mtDNA pathological variants were present at similarly low levels. As per previous studies we found increased pathogenic variants within the mtDNA control region (D-Loop). However overall, there was no significant difference in the distribution of variants across the rest of the genome in all patients. Although a higher proportion of non-synonymous variants were seen this was not statistically significant.</p> <p>We have shown that it is possible to isolate and sequence individual MSCs and observe mtDNA pathological variants. This gives a snapshot in time of the variant load for each patient. Due to low sample numbers compared to other studies investigate mtDNA in other cell types, we did not observe any age related increase in pathological variants. However this adds further evidence that clonally expanded, somatic mtDNA variants are common and could contribute to age related disease including osteoporosis.</p>
Raw gel and membrane figures associated with the publication "Iron-loaded deferiprone can support full hemoglobinization of cultured red blood cells"
<p>Raw gel and membrane figures associated with the publication <strong>Iron-loaded deferiprone can support full hemoglobinization of cultured red blood cells</strong>.</p> <p> </p> <p><strong>For Figure 1a:</strong></p> <p>Raw uncropped image of the gel run with samples combining apotransferrin with iron chelators. After staining with Coomassie, gel was scanned in an Epson V500 scanner (full color, 1200 dpi, bit depth = 24).</p> <p> </p> <p><strong>For Figure 3:</strong></p> <p>Raw uncropped images of the membranes obtained from western blots of cultured erythroblasts derived from three different donors, showing the expression level of proteins involved in iron metabolism regulation (ferritin, transferrin receptor, total EIF2, phospho-EIF2). Actin was used as housekeeping protein for relative quantification of protein abundance for further analysis.</p> <p>After staining, membranes were scanned in an Epson V500 scanner (full color, 1200 dpi, bit depth = 24).</p> <p>For all membranes, the loading order is the same:</p> <p>- <strong>Day 0:</strong> 1000 μg/mL holotransferrin</p> <p>- <strong>Day 0:</strong> 100 μg/mL holotransferrin</p> <p>- <strong>Day 0:</strong> 100 μg/mL holotransferrin + 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 1:</strong> 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 1:</strong> 1000 μg/mL apotransferrin</p> <p>- <strong>(empty lane)</strong></p> <p>- <strong>Day 1:</strong> 1000 μg/mL holotransferrin</p> <p>- <strong>Day 1:</strong> 100 μg/mL holotransferrin</p> <p>- <strong>Day 1:</strong> 100 μg/mL holotransferrin + 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 1:</strong> 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 1:</strong> 1000 μg/mL apotransferrin</p> <p>- <strong>(empty lane)</strong></p> <p>- <strong>Day 2:</strong> 1000 μg/mL holotransferrin</p> <p>- <strong>Day 2:</strong> 100 μg/mL holotransferrin</p> <p>- <strong>Day 2:</strong> 100 μg/mL holotransferrin + 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 2:</strong> 52 μM iron-loaded deferiprone</p> <p>- <strong>Day 2:</strong> 1000 μg/mL apotransferrin</p> <p>More details on the protocol for western blot available in the manuscript at https://doi.org/10.1101/2021.08.02.454758</p> <p> </p> <p><strong>For Figure 4:</strong></p> <p>Raw uncropped images of the membranes obtained from western blots of cultured erythroblasts derived from two different donors with different iron supplementations (no hTf or Def, 52 μM iron-loaded deferiprone, or 300 μg/mL holotransferrin), showing the phosphorylation level of STAT5 upon withdrawal of erythropoietin (Epo) (3 hours) and restimulation with 0.2 or 1.0 U/mL Epo.</p> <p>After staining, membranes were scanned in an Epson V500 scanner (full color, 1200 dpi, bit depth = 24).</p> <p>For both membranes, the loading order is the same:</p> <p>- <strong>Lane 1:</strong> cells cultured without hTf nor Def, and no Epo restimulation</p> <p>- <strong>Lane 2:</strong> cells cultured with 52 μM iron-loaded deferiprone, and no Epo restimulation</p> <p>- <strong>Lane 3:</strong> cells cultured with 300 μg/mL holotransferrin, and no Epo restimulation</p> <p>- <strong>Lane 4:</strong> cells cultured without hTf nor Def, and restimulation with 0.2 U/mL Epo.</p> <p>- <strong>Lane 5:</strong> cells cultured with 52 μM iron-loaded deferiprone, and restimulation with 0.2 U/mL Epo.</p> <p>- <strong>Lane 6:</strong> cells cultured with 300 μg/mL holotransferrin, and restimulation with 0.2 U/mL Epo.</p> <p>- <strong>Lane 7:</strong> cells cultured without hTf nor Def, and restimulation with 1.0 U/mL Epo.</p> <p>- <strong>Lane 8:</strong> cells cultured with 52 μM iron-loaded deferiprone, and restimulation with 1.0 U/mL Epo.</p> <p>- <strong>Lane 9:</strong> cells cultured with 300 μg/mL holotransferrin, and restimulation with 1.0 U/mL Epo.</p>
Removal of senescent cells reduces the viral load and attenuates pulmonary and systemic inflammation in SARS-CoV-2-infected, aged hamsters
<p>Older age is one of the strongest risk factors for coronavirus disease 2019 (COVID-19) morbidity and mortality. In an older adult, the chronic accumulation of senescent cells can interfere with the immune system and accentuate inflammation. Here, we sought to determine whether age-associated cellular senescence contributes to the severity of COVID-19 by studying the well-established golden hamster model of severe acute respiratory syndrome coronavirus (SARS-CoV-2)-driven lung disease. We found that aged hamsters (22 months of age) accumulate senescent cells in the lungs and that the senolytic drug ABT-263 (a Bcl-2-family inhibitor) depletes these cells at baseline and during a SARS-CoV-2 infection (when the senescent cell count is typically elevated). Relative to young hamsters (2 months of age), aged hamsters had a greater viral load during the acute phase of infection and displayed higher levels of fibrosis and worse body weight recovery during the post-acute phase. Interestingly, early treatment with ABT-263 was associated with a significantly lower pulmonary viral load and an amelioration of COVID-19-like lung disease in aged (but not young) animals. ABT-263 treatment of aged animals was also associated with lower pulmonary and systemic levels of senescence-associated secretory phenotype factors. We conclude that the removal of senescent cells via treatment with a senolytic reduces the pathologic severity of SARS-CoV-2 infection in aged hamsters. As several senolytics have recently moved into early-stage clinical trials, our present findings have clear clinical relevance.</p>
Dynamic interactome of the MHC I peptide loading complex in human dendritic cells - Source Ib
<p>Source data underlying MS data set (Fig.1). Datafile comprises MaxQuant output files.</p>
Vaccination of Melanoma Patients With Dendritic Cells Loaded With Allogeneic Apoptotic-Necrotic Melanoma Cells
ClinicalTrials.gov study NCT00515983. IPD Sharing: Not stated. Countries: 1. Publications: 1.
IKKb-matured, RNA-loaded Dendritic Cells for Metastasised Uveal Melanoma
ClinicalTrials.gov study NCT04335890. IPD Sharing: NO. Countries: 1. Publications: 4.
Dendritic Cells Loaded With Allogeneous Cell Lysate in Mesothelioma Patients
ClinicalTrials.gov study NCT02395679. IPD Sharing: Not stated. Countries: 1. Publications: 6.
Efficiency of Vaccination with Lysate-loaded Dendritic Cells in Patients with Newly Diagnosed Glioblastoma
ClinicalTrials.gov study NCT03395587. IPD Sharing: NO. Countries: 1. Publications: 2.
Cellular Immunotherapy Study With Autologous Dendritic Cells Loaded With Oncofetal Antigen/iLRP in Patients With Metastatic Breast Cancer
ClinicalTrials.gov study NCT00879489. IPD Sharing: Not stated. Countries: 1. Publications: 17.
Autologous Dendritic Cells Loaded With Autologous Tumor Associated Antigens for Treatment of Newly Diagnosed Glioblastoma
ClinicalTrials.gov study NCT03400917. IPD Sharing: Not stated. Countries: 1. Publications: 1.
Carcinoembryonic Antigen-loaded Dendritic Cells in Advanced Colorectal Cancer Patients
ClinicalTrials.gov study NCT00228189. IPD Sharing: Not stated. Countries: 1. Publications: 5.
Treatment of Infertility by Collagen Scaffold Loaded With Umbilical Cord Derived Mesenchyma Stem Cells
ClinicalTrials.gov study NCT02313415. IPD Sharing: NO. Countries: 1. Publications: 1.
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OpenNeuro
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