title stringlengths 1 1.19k | keywords stringlengths 0 668 | concept stringlengths 0 909 | paragraph stringlengths 0 61.8k | PMID stringlengths 10 11 |
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Statistical analysis | dysplasia | ESOPHAGEAL CANCER, DYSPLASIA | Summary statistics for continuous variables (means and standard deviations) and categorical variables (counts and proportions) were presented for both the intervention and control groups. An aggregate of the compliance for all 12 weeks for each individual was obtained by dividing 100/12 for example, 100% compliance if ... | PMC9939163 |
RESULTS | PMC9939163 | |||
Study participants | esophageal cancer, D) Muscle mass | OESOPHAGEAL CANCER | Patient enrolment was consecutive and lasted from January 2016 to August 2020. A total of 243 survivors from the OSCAR study were assessed for eligibility. Of them, 233 met the inclusion criteria, but 72 declined to participate. The remaining 161 participants (69.1% of those eligible) were randomly assigned to either t... | PMC9939163 |
Hand grip strength | esophageal cancer | OESOPHAGEAL CANCER | Among the 70 patients with completed measures in the control group, two patients had missing values for hand grip strength and hence were not included in the final analysis (Figure Differences in primary outcomes between baseline and post‐intervention for the two allocation groups of the Swedish nationwide randomized c... | PMC9939163 |
Lower extremity strength | From those who had completed measures in the control group ( | PMC9939163 | ||
Muscle mass | Among the participants with complete end of study measures in the intervention group ( | PMC9939163 | ||
DISCUSSION | esophageal cancer, lower extremity muscle strength, muscle mass | ESOPHAGEAL CANCER, RECRUITMENT, OESOPHAGEAL CANCER | This trial demonstrated that a specially designed 12‐week home‐based PA program improved lower extremity muscle strength among patients having undergone surgery for esophageal cancer 1‐year earlier, while no influence on hand grip strength or muscle mass was identified.Some methodological strengths and weaknesses of th... | PMC9939163 |
FUNDING INFORMATION | ALF, Cancer | CANCER | This work is supported by the Swedish Cancer Society 180685, the Swedish Research Council 2017–01744, the Swedish Research Council for Health, Workinglife and Welfare 2017–00812, The Cancer Research Funds of Radiumhemmet 171103 and Region Stockholm (ALF) LS 2018–1157. Pernilla Lagergren is supported by the NIHR Imperia... | PMC9939163 |
CONFLICT OF INTEREST | The authors declare no conflict of interest. | PMC9939163 | ||
ETHICAL APPROVAL STATEMENT | Ethical approval was granted by the Regional Ethical Review Board in Stockholm, Sweden (Dnrs: 2015/2142–32; 2016/1696–32/1; 2018/1447–32). | PMC9939163 | ||
CONSENT TO PARTICIPATE | Written and oral information was given, and informed consent was collected from all participants by the research nurse before the randomization envelope was opened. | PMC9939163 | ||
TRIAL REGISTRATION | PMC9939163 | |||
ACKNOWLEDGMENTS | We acknowledge patients of the PA study for their valuable participation and our Surgical Care Science patient research partnership group for valuable inputs on the study. | PMC9939163 | ||
DATA AVAILABILITY STATEMENT | The data sets generated and/or analyzed in the current study will not be publicly available due to the ethical review act but will be available from the principal investigators of the study on reasonable request. | PMC9939163 | ||
REFERENCES | PMC9939163 | |||
2. Materials and Methods | PMC10346321 | |||
2.1. Study Design and Participants | chronic non-communicable diseases, amenorrhea | VASODILATION, CARDIOVASCULAR DISEASES | A randomized trial conducted on 48 individuals compared the effects of resveratrol supplementation (500 mg/day) and energy restriction (1000 kcal/day) for 30 days on SIRT1 and vascular reactivity parameters. The study participants were 24 postmenopausal women (01 years of natural amenorrhea) and 24 men aged 55 to 65, a... | PMC10346321 |
2.2. Biochemical Analysis | Laboratory tests were performed with biological samples collected after a 12-h fast. Serum samples were obtained after collecting venous blood in tubes without an anticoagulant and centrifuged for 20 min at 1800G (Eppendorf, Hamburg, Germany). Citrated blood samples were centrifuged for 10 min at 200× | PMC10346321 | ||
2.3. Sirtuin 1 Assessment | SIRT1 serum concentration was determined using an ELISA kit (Uscn Life Science, Wuhan, Hubei, China). Serum samples, before and after interventions, were analyzed in duplicate and in the same ELISA plate using the Multiscan FC plate reader (Thermo Scientific, Waltham, MA, USA), with a coefficient of variation of 12%, a... | PMC10346321 | ||
2.4. Resveratrol Purity and Formulation Analysis | The resveratrol administered to participants was obtained from a compounding pharmacy (Buenos Aires Pharmacy, São Paulo, Brazil). The purity of the product supplied was analyzed by capillary electrophoresis using a Proteome Lab PA800 (Beckman Coulter, Fullerton, CA, USA) at the Laboratory of Capillary Chromatography an... | PMC10346321 | ||
2.5. Vascular Reactivity Assessment | Endothelium-dependent flow-mediated vasodilation | VASODILATION | Endothelium-dependent flow-mediated vasodilation (FMD) and endothelium-independent vasodilation (NMD) were assessed according to a previous guideline [ | PMC10346321 |
2.6. Statistical Analyses | REGRESSIONS | The sample size of 48 patients, with 24 subjects per treatment arm, was determined to yield a power of 80% with a 5% significance level to detect a 30% difference in Sirt1 serum concentrations. Participants were randomly assigned in a 1:1 ratio using computer-generated numbers to include participants in the resveratrol... | PMC10346321 | |
3. Results | REGRESSIONS | Baseline and post-intervention data are described in At the end of the study, individuals in the energy restriction group had a significant reduction in weight (When comparing groups, there was a statistically significant difference between the changes (∆) in weight (Both interventions increased serum SIRT1 (Correlatio... | PMC10346321 | |
4. Discussion | cerebral ischemic stress | VASODILATION, CARDIOVASCULAR DISEASES, INFLAMMATION, OXIDATIVE STRESS, ENDOTHELIAL DYSFUNCTION | The main results of our study showed that 30 days of energy restriction, i.e., a 1000 kcal diet, or 500 mg daily resveratrol supplementation similarly increased circulating SIRT1 and decreased plasma noradrenaline. Plasma noradrenaline reduction was correlated with higher baseline circulating SIRT1. After the intervent... | PMC10346321 |
Author Contributions | Conceptualization, A.d.P.M. and G.H.F.G.; Methodology, A.d.P.M.; Validation, A.d.P.M.; Data collection, A.R., M.F.d.S.G., S.D.A., C.M.C.S. and A.d.P.M.; Data analysis, A.d.P.M. and G.H.F.G.; Writing—Original draft preparation, A.d.P.M., G.H.F.G., K.L.K. and N.F.d.O.F.; Writing—review and editing, A.d.P.M. and G.H.F.G.;... | PMC10346321 | ||
Institutional Review Board Statement | HEART | The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of Heart Institute of University of São Paulo Medical School (protocol code CAAE:00788012.8.0000.0068). | PMC10346321 | |
Informed Consent Statement | Informed consent was obtained from all subjects involved in the study. | PMC10346321 | ||
Data Availability Statement | Not applicable. | PMC10346321 | ||
Conflicts of Interest | The authors declare no conflict of interest. | PMC10346321 | ||
References | AD | VASODILATION | Research protocol design.Clinical and biochemical characteristics of the participants at baseline and post-treatment.Data are presented as median (interquartile range—IQR). *: Comparison of intra-groups post–pre-tests. **: Comparison between groups at baseline. Significant values are bold.Comparison of the median diffe... | PMC10346321 |
1. Introduction | Obesity, obesity, death, breast cancer, Breast cancer, Smartphone-Based Cancer, deaths, Cancer | OBESITY, OBESITY, BREAST CANCER, BREAST CANCER, SECONDARY, CANCER | Breast cancer prevalence has increased globally, with 12.2% of breast cancer cases identified in China. Obesity and unhealthy lifestyles are major risk factors for breast cancer. We conducted a randomized control trial to assess the feasibility and evaluate the preliminary effect of the Smartphone-Based Cancer and Obes... | PMC10218157 |
2. Materials and Methods | DISEASE, RECRUITMENT | This pilot study used a randomized control trial design. The social cognitive theory (SCT) [Women who met the inclusion criteria from two community health centers in Changsha, Hunan Province, China, were invited to this study and randomized to either the SCOPE or control programs. Inclusion criteria included (1) being ... | PMC10218157 | |
2.1. Intervention Contents | Obesity, WeChat-based, Cancer | OBESITY, BREAST CANCER, DISEASE, CANCER | Fitbit tracker. All participants received a Fitbit Alta HRTwelve modules. Modules were sent via WeChat. Participants received 12 weekly, culturally appropriate, evidence-based educational modules along with tailored tips and messages via WeChat. Each module includes an educational session/information that lasts less th... | PMC10218157 |
2.2. Control Group | The control group received a Fitbit Alta HR | PMC10218157 | ||
2.3. Data Collection and Measures | obesity, Diabetes | OBESITY, BREAST CANCER, CREST, DIABETES | Data were collected at baseline, 3 months, and 6 months. The research assistants and a registered nurse conducted all measurements. The trained research assistant and registered nurse measured WC, weight, and height. Study participants completed all questionnaires in a separate room where a research assistant could ans... | PMC10218157 |
2.4. Data Analysis | Because this is a pilot study aimed at testing the feasibility, acceptance, and preliminary efficacy, a power analysis was not conducted. Since this pilot RCT has a limited sample size, analyses were intended to be exploratory, and we set α = 0.05 (two-sided). Descriptive statistics (means and standard deviations, or m... | PMC10218157 | ||
3. Results | PMC10218157 | |||
3.1. Sample Characteristics | A total of 102 women (52 in the SCOPE intervention and 50 in the control group) participated in this study (see | PMC10218157 | ||
3.2. Effect of the Intervention | Results from the mixed-model analysis found that at the 3-month follow-up, compared to baseline, participants in the SCOPE program compared to control significantly reduced WC (At 6 months, compared to baseline, women in the SCOPE group compared to control significantly reduced WC ( | PMC10218157 | ||
3.3. Feasibility and Satisfaction of the SCOPE Modules | A total of 85% of participants completed 6 months of follow-up. The main reasons for missing the follow-up assessment include lack of time and inability to come to the site for assessment. The average number of modules viewed was 8.5 out of 12 modules. Most participants in the SCOPE intervention program (93%) indicated... | PMC10218157 | ||
4. Discussion | obesity, abdominal obesity, breast cancer | OBESITY, SDH, BREAST CANCER | As breast cancer is a significant health issue, and obesity increases the risk for breast cancer, especially for women with children, we developed a smartphone-based intervention using a popular communication app tailored to Chinese mothers. Our intervention aimed at reducing obesity, increasing knowledge and attitude,... | PMC10218157 |
5. Conclusions | obesity, breast cancer | OBESITY, BREAST CANCER | This pilot study demonstrates a promising way to prevent obesity and breast cancer. The SCOPE intervention delivered via a communication app is feasible and may positively reduce women’s risk for obesity and improve breast cancer knowledge and attitude. This intervention has great potential to promote the health and we... | PMC10218157 |
Author Contributions | Conceptualization, J.-L.C. and J.G.; methodology, J.-L.C.; software, T.J.H. and C.-X.L.; validation, J.-L.C., J.G. and T.J.H.; formal analysis, T.J.H., J.-L.C. and C.-X.L.; investigation, Q.Z., Y.J., P.M., H.Z. and Q.H.; resources, P.M., H.Z. and Q.H.; data curation, C.-X.L., Q.Z., Y.J. and Q.H.; writing—original draft... | PMC10218157 | ||
Institutional Review Board Statement | The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Ethics Committee on Human Research of the University of California San Francisco (IRB 18-27025). | PMC10218157 | ||
Informed Consent Statement | Informed consent was obtained from all subjects involved in the study. | PMC10218157 | ||
Data Availability Statement | The data presented in this study are available on request from the corresponding author. The data are not publicly available due to them containing personal information. | PMC10218157 | ||
Conflicts of Interest | The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. | PMC10218157 | ||
References | RECRUITMENT | Sample session in WeChat. Patient recruitment flow chart.Demographics.Baseline data.Comparisons between control and SCOPE experimental groups.Mixed model and effect size of the intervention. | PMC10218157 | |
Visual Abstract | breast cancer | METASTATIC DISEASE, BREAST CANCER | Published online Jul. 13, 2023.
Imaging using the human epidermal growth factor receptor 2 (HER2)–binding tracer Up to 20% of breast cancer cases have human epidermal growth factor receptor 2 (HER2) overexpression with or without HER2 oncogene amplification (Currently, treatment failure is relatively common, and cure r... | PMC10478820 |
MATERIALS AND METHODS | PMC10478820 | |||
Patient Population | tumor, PBC, neutropenia | TUMOR, HER2-POSITIVE BREAST CANCER, PBC, NEUTROPENIA, MALIGNANCIES, CONGESTIVE HEART FAILURE, CONCOMITANT DISEASE, HER2-NEGATIVE BREAST CANCER | The current study was a planned interim analysis of a phase II study embedded within a larger academically driven prospective open-label phase II and phase III diagnostic trial, approved by the Swedish Medical Products Agency (EudraCT 2017-002115-34; diarienummer, 5.1-2018-30296; Women with newly diagnosed stage II or ... | PMC10478820 |
Production of [ | The ABY-025 peptide was provided by Affibody AB. Production of [ | PMC10478820 | ||
[ | All scans were performed using a Discovery MI PET/CT scanner (GE Healthcare), and scans included a section from the skull apex to mid thigh. [The first 10 patients were closely monitored by electrocardiograms and clinical examinations during their stay at the PET center. All patients received a phone call after 24 h fr... | PMC10478820 | ||
[ | [ | PMC10478820 | ||
PET Measurements | Images obtained from both [ | PMC10478820 | ||
Biopsies | Lesions were selected for study biopsies on the basis of uptake on both PET scans and accessibility at a multidisciplinary trial conference the day after the last baseline PET scan. The biopsied lesions were carefully located and manually defined on [ | PMC10478820 | ||
HER2-Targeted Treatment | multiple recurrent disease | RECURRENT DISEASE | Patients planned for neoadjuvant treatment and patients with first-time recurrent disease received trastuzumab, pertuzumab, and chemotherapy according to guidelines, whereas patients with multiple recurrent disease received trastuzumab emtansine. | PMC10478820 |
Statistical Analysis | PET metrics were reported as mean ± SD unless otherwise stated. Receiver operating characteristic curve analysis was used to investigate the predictive value of the variables and to define a cutoff for [ | PMC10478820 | ||
RESULTS | PMC10478820 | |||
Patient Characteristics | RECRUITMENT, BREAST CANCER | Patient characteristics and descriptive data are shown in Patient Characteristics and Descriptive DataBased on immunohistochemistry and ISH results.Qualitative data are number and percentage; continuous data are median and range.Anatomic Distribution and [Number of biopsied lesions is in parentheses, 1 per patient. SUV... | PMC10478820 | |
Tumor Lesions | We measured tracer uptake in up to 5 of the largest lesions per patient, including the biopsied lesion. Anatomic distribution of measured lesions is shown in [ | PMC10478820 | ||
[ | In total, 12 patients showed a mismatch between PET and HER2 status, using SUV | PMC10478820 | ||
Clinical and Metabolic Response | therapy-naïve, PBC, breast cancer | DISEASE, PBC, BREAST CANCER | Thirty-two patients had either partial or complete metabolic response after receiving 2 cycles of treatment. Clinically, 22 patients had complete response, 11 had partial response, 2 had stable disease, and 5 had progressive disease. Global Δ-TLG was significantly associated with the clinical response ([Receiver operat... | PMC10478820 |
DISCUSSION | coronavirus disease 2019 | CORONAVIRUS DISEASE 2019 | The potential of [The results showed that immunohistochemistry staining did not always reflect the biologic availability of the receptors; a HER2-positive biopsy sample with low [We found an inverse association between the number of previous treatments and the metabolic response to current treatment. The more treatment... | PMC10478820 |
CONCLUSION | BREAST CANCER | Immunohistochemistry staining and ISH are currently the gold standards to determine HER2 status in breast cancer. However, limitations in the metastatic setting still hinder accurate biopsy-based evaluation of heterogeneous HER2 expression. Hence, the advantage of noninvasive techniques such as [ | PMC10478820 | |
DISCLOSURE | Breast Cancer, Cancer | BREAST CANCER, CANCER | This work was partially supported by grants from the Swedish Breast Cancer Association, Swedish Cancer Foundation (19 0507 Pj), Roche AB Sweden, and the Percy Falk Foundation. Fredrik Frejd and Joachim Feldwisch are employees and own shares in Affibody AB. Jens Sörensen received clinical advisor remunerations from Affi... | PMC10478820 |
REFERENCES | PMC10478820 | |||
Subject terms | AML, cancers, myelofibrosis | CANCERS, ACUTE MYELOID LEUKEMIA, AML, MYELOFIBROSIS, MUTANT, MYELODYSPLASTIC SYNDROMES | Telomerase enables replicative immortality in most cancers including acute myeloid leukemia (AML). Imetelstat is a first-in-class telomerase inhibitor with clinical efficacy in myelofibrosis and myelodysplastic syndromes. Here, we develop an AML patient-derived xenograft resource and perform integrated genomics, transc... | PMC10824665 |
Main | blood cancer, AML | BLOOD CANCER, DISEASE, AML, LEUKEMIA STEM CELL | AML is an aggressive and lethal blood cancer with a 5-year overall survival rate of less than 45% for patients younger than 60 years of age and less than 10% for older patients, predominantly due to disease relapse after chemotherapy or targeted treatments. AML has been extensively classified based on biological featur... | PMC10824665 |
Results | PMC10824665 | |||
Generation of a comprehensive AML PDX resource | AML, thrombocytopenia, anemia, splenomegaly, AML PDX | DISEASE, THROMBOCYTOPENIA, AML, ANEMIA | To generate a representative AML PDX inventory, primary bone marrow or blood samples from 50 patients were tested for engraftment and development of AML in NOD/SCID/IL2gR−/−/hIL3,CSF2,KITLG (NSGS). The overall success rate for primary engraftment in NSGS was 70%, defined by bone marrow, spleen or peripheral blood donor... | PMC10824665 |
A phase II-like preclinical trial of imetelstat in AML PDX | AML | AML | To test the preclinical efficacy of imetelstat in AML, the characterized 30 individual samples from patients with AML were each transplanted into 12 NSGS recipients ( | PMC10824665 |
A CRISPR/Cas9 screen to identify key effectors of imetelstat | AML | To investigate the mechanism of action of imetelstat in AML in an unbiased manner, we applied the Brunello guide RNA (gRNA) library | PMC10824665 | |
Lipophagy precedes imetelstat-induced ferroptosis | AML | By integrating transcriptomics and functional genetics, we aimed to investigate the mechanism by which imetelstat induces ferroptosis. We performed an overlay of the in vivo AML PDX RNA-seq datasets from imetelstat and vehicle-treated mice with the Brunello library CRISPR/Cas9 knockout screen data (cutoff criteria of R... | PMC10824665 | |
Oxidative stress signatures distinguish sustained responders | AML, hematological malignancies | DISEASE, AML, HEMATOLOGICAL MALIGNANCIES | We next aimed to identify biomarkers of imetelstat response and resistance. Improved survival in imetelstat-treated AML PDXs correlated with significantly reduced engraftment and disease burden; however, there were clear differences in the magnitude and duration of individual responses (Extended Data Fig. We next aimed... | PMC10824665 |
Oxidative stress induction sensitizes AML PDX to imetelstat | OXIDATIVE STRESS | The finding that responses to imetelstat are associated with baseline molecular signatures annotated as oxidative stress and that the mechanism of action of imetelstat features ROS-mediated ferroptosis led to the hypothesis that oxidative stress induction can sensitize to imetelstat therapy.Standard induction chemother... | PMC10824665 | |
Discussion | AML, death, hematological myeloid malignancies, myelofibrosis | ESSENTIAL THROMBOCYTHEMIA, OXIDATIVE STRESS, AML, MYELOFIBROSIS | Imetelstat is a first-in class telomerase inhibitor with clinical efficacies in hematological myeloid malignancies, including essential thrombocythemia, myelofibrosis and lower-risk myelodysplastic syndromesFerroptosis is a recently discovered type of non-apoptotic regulatory cell death that relies on the balance of th... | PMC10824665 |
Methods | Our research complies with all relevant ethical regulations, including QIMR Berghofer human research ethics committee protocol P1382 (HREC/14/QRBW/278) and QIMR Berghofer animal research ethics committee protocol A11605M. Animals were monitored daily and immediately killed based on the scoring criteria detailed below. | PMC10824665 | ||
Mouse monitoring | weight loss, hunching, paralysis | Animals were monitored daily and always immediately killed as soon as a cumulative clinical score of 3 or above was reached, based on weight loss (score 1, >10–20%; and score 2, >20% or >15% if maintained for >72 h), posture (score 1, hunching noted only at rest; and score 2, severe hunching), activity (score 1, mild t... | PMC10824665 | |
Mouse models | All mouse experiments were approved by the institutional (QIMR Berghofer) ethics committee protocol A11605M. NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjz/SzJ), NSGS (NOD.Cg-Prkdcscid Il2rgtm1Wjl Tg[CMV-IL3,CSF2,KITLG]1Eav/MloySzJ) and NRGS (NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl Tg[CMV-IL3,CSF2,KITLG]1Eav/J) were imported from Jackson Lab... | PMC10824665 | ||
Xenograft transplantation experiments | AML | AML | AML samples were obtained from patients, after informed consent in accordance with the Declaration of Helsinki. Ficoll density gradient was used to recover viable mononuclear cells. Viably frozen AML cells were thawed and CD3-depleted with biotinylated anti-human CD3 (SK7) and biotin-binder Dynabeads (Invitrogen) and s... | PMC10824665 |
Oligonucleotide sequences of imetelstat and mismatch controls | Imetelstat (GRN163L): 5′ R-TAGGGTTAGACAA-NH2 3′.Mismatch 1 (GRN140833): 5′ R-TAGGTGTAAGCAA-NH2 3′.Mismatch 2 (GRN142865): 5′ R-TAGGGATTCAGAA-NH2 3′. | PMC10824665 | ||
Drug treatment studies | NSG, NSGS or NRGS mice were treated with 15 mg kg | PMC10824665 | ||
Blood analysis | BLOOD | Blood was collected into EDTA-coated tubes and analyzed on a Hemavet 950 (Drew Scientific). PB smears were stained with Wright-Giemsa according to the manufacturer’s protocol (BioScientific). | PMC10824665 | |
Histology | Tissues were fixed in 10% neutral buffered formalin, embedded in paraffin and stained with hematoxylin and eosin. Images of histological slides were obtained on a ScanScope FL (Aperio). | PMC10824665 | ||
Flow cytometry analysis of AML PDX and cord blood transplants | AML | LYSIS, AML | For monitoring AML engraftment, 25–50 μl of PB were stained after red cell lysis (BD Pharm Lyse, BD Biosciences) with anti-human CD45-AF647 (H130) and anti-mouse CD45.1-PE (A20). For AML phenotyping, cell populations were purified from bone marrow (both femurs and tibiae) or SPL and after red blood cell lysis stained w... | PMC10824665 |
Terminal restriction fragment analysis | TRFs were obtained from genomic DNA by complete digestion with the restriction enzymes HinfI and RsaI. TRFs were separated by pulsed-field gel electrophoresis. Gels were dried, denatured and subjected to in-gel hybridization with a γ-[32P]-ATP-labeled (CCCTAA)4 oligonucleotide probe. Gels were washed and the telomeric ... | PMC10824665 | ||
Telomere length qPCR | BLOOD | Samples were purified using the DNeasy Blood and Tissue kit (QIAGEN). DNA isolation was performed as described previously | PMC10824665 | |
Cell lines | The | PMC10824665 | ||
Cell culture and in vitro cell growth analysis | AML | AML | AML cell lines were cultured in RPMI with 10% fetal calf serum, 2 mM glutamine and 200 U ml | PMC10824665 |
Imaging flow cytometry | Lipophagy was detected by assessing colocalization of C12-BODIPY and LAMP1 using a previously published method with modifications | PMC10824665 | ||
Flow cytometry analysis of AML cell lines | Before staining, 2 × 10Flow cytometry analysis of lipid peroxidation was performed using C11-BODIPY 581/591 (Sapphire Bioscience) according to a previously published protocol | PMC10824665 | ||
Western blotting | LYSED, SECONDARY, LYSIS | Cells were lysed on ice using m-PER lysis buffer (Thermo Fisher Scientific) supplemented with a protease and phosphatase inhibitor (Cell Signaling, 5872) and protein was quantified using Pierce BCA protein assay kit (Thermo Fisher Scientific). In total, 20–50 μg of protein extract was electrophoresed on a 4–15% SDS gra... | PMC10824665 | |
Confocal microscopy | FLUOR, COLD | Cytospins were fixed and permeabilized with methanol:acetone (prechilled) at a ratio of 1:1 for 10 min at room temperature, then washed twice with cold PBS and once with room temperature PBS (5 min each). Cytospins were then incubated with 1% BSA–PBS at room temperature for 1 h, washed three times in PBS for 5 min each... | PMC10824665 | |
CRISPR/Cas9 editing | The Brunello genome-wide gRNA library contains 76,441 gRNAs targeting 19,114 genes and was obtained from Addgene (cat. 73178) | PMC10824665 | ||
Mutational sequencing | cancer | CANCER, HEMATOLOGICAL MALIGNANCIES | Genomic alterations were profiled using the HemePACT assay (integrated mutation profiling of actionable cancer targets related to hematological malignancies) | PMC10824665 |
RNA sequencing | RNA was isolated from a maximum of 0.5 × 10 | PMC10824665 | ||
Lipidomics | Targeted lipidomics was performed on a 1290 Infinity II UHPLC coupled to a 6470 QQQ mass spectrometer via AJS ESI source (Agilent) in positive ionization mode, using a scheduled multiple reaction monitoring (MRM) method | PMC10824665 | ||
Statistical analyses | Unless otherwise stated, statistical analyses were carried out using GraphPad Prism v.9.4.0. Microsoft Excel for Mac v.16.75.2 was used to re-calculate those | PMC10824665 | ||
Statistics and reproducibility | Study design was based on sample sizes that proved to be adequate in previous experiments using similar approaches and thus no statistical methods were used to predetermine sample sizes for this study | PMC10824665 |
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