Datasets were obtained from the GEO database through the following GEO Series accession numbers: GSE93391, GSE114012, GSE131594, GSE152699, GSE124854, GSE135215, GSE99116, GSE124109, GSE61130, GSE64553 and GSE63577.
Cell lines were obtained from ATCC or Sigma and regularly checked for mycoplasma. A549 and NCIH460 were cultured in DMEM . NCIH358, NCIH1299 and NCIH1563 were maintained in RPMI-1640 supplemented with 5 mM sodium pyruvate and 0.5% glucose. NCIH1944, NCIH1666, NCIH1650 and L23 were grown in RPMI-1640 ATCC formulation . A427 were cultured in EMEM . A549, NCIH460, H358, NCIH1299, NCIH1563, and A427 were supplemented with 10% heat inactivated FBS.
Endogenous PCNA was labelled at the N-terminus with a cDNA encoding mRuby in both A549 and NCI-H1944 cells, using AAV-mediated gene-targeting, according to methods described in Zerjatke et al. []. mRuby-expressing cells were sorted into 50:50 conditioned:fresh media at single-cell density into 96-well plates by FACS and single-cell clones expanded. For live-cell imaging, 500 cells in phenol-red free media were plated per well of a 384 CellCarrierUltra plate the day before imaging.
The G0 arrest scores for cancer cell lines were calculated using corresponding log-transformed RPKM normalised bulk RNA-seq data from the Cancer Cell Line Encyclopedia database [CEP89 was depleted by ON-Target siRNA Pool from Horizon. NCI-H1299 cells were reverse transfected in 384-well plates with 20 nM of non-targeting control or CEP89-targeting siRNA using Lipofectamine RNAiMax , according to the manufacturer’s instructions.
The mutation rates of all TCGA primary tumour samples were determined by log-transforming the total number of mutations in each sample divided by the length of the exome capture .functional status was assessed based on somatic mutation and copy number alterations as described in Zhang et al. [
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Analysis of the microglia transcriptome across the human lifespan using single cell RNA sequencing - Journal of NeuroinflammationBackground Microglia are tissue resident macrophages with a wide range of critically important functions in central nervous system development and homeostasis. Method In this study, we aimed to characterize the transcriptional landscape of ex vivo human microglia across different developmental ages using cells derived from pre-natal, pediatric, adolescent, and adult brain samples. We further confirmed our transcriptional observations using ELISA and RNAscope. Results We showed that pre-natal microglia have a distinct transcriptional and regulatory signature relative to their post-natal counterparts that includes an upregulation of phagocytic pathways. We confirmed upregulation of CD36, a positive regulator of phagocytosis, in pre-natal samples compared to adult samples in situ. Moreover, we showed adult microglia have more pro-inflammatory signature compared to microglia from other developmental ages. We indicated that adult microglia are more immune responsive by secreting increased levels of pro-inflammatory cytokines in response to LPS treatment compared to the pre-natal microglia. We further validated in situ up-regulation of IL18 and CXCR4 in human adult brain section compared to the pre-natal brain section. Finally, trajectory analysis indicated that the transcriptional signatures adopted by microglia throughout development are in response to a changing brain microenvironment and do not reflect predetermined developmental states. Conclusion In all, this study provides unique insight into the development of human microglia and a useful reference for understanding microglial contribution to developmental and age-related human disease.
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Prevalence and risk factors of significant persistent pain symptoms after critical care illness: a prospective multicentric study - Critical CareBackground Prevalence, risk factors and medical management of persistent pain symptoms after critical care illness have not been thoroughly investigated. Methods We performed a prospective multicentric study in patients with an intensive care unit (ICU) length of stay ≥ 48 h. The primary outcome was the prevalence of significant persistent pain, defined as a numeric rating scale (NRS) ≥ 3, 3 months after admission. Secondary outcomes were the prevalence of symptoms compatible with neuropathic pain (ID-pain score | 3) and the risk factors of persistent pain. Results Eight hundred fourteen patients were included over a 10-month period in 26 centers. Patients had a mean age of 57 (± 17) years with a SAPS 2 score of 32 (± 16) (mean ± SD). The median ICU length of stay was 6 [4–12] days (median [interquartile]). At 3 months, the median intensity of pain symptoms was 2 [1–5] in the entire population, and 388 (47.7%) patients had significant pain. In this group, 34 (8.7%) patients had symptoms compatible with neuropathic pain. Female (Odds Ratio 1.5 95% CI [1.1–2.1]), prior use of anti-depressive agents (OR 2.2 95% CI [1.3–4]), prone positioning (OR 3 95% CI [1.4–6.4]) and the presence of pain symptoms on ICU discharge (NRS ≥ 3) (OR 2.4 95% CI [1.7–3.4]) were risk factors of persistent pain. Compared with sepsis, patients admitted for trauma (non neuro) (OR 3.5 95% CI [2.1–6]) were particularly at risk of persistent pain. Only 35 (11.3%) patients had specialist pain management by 3 months. Conclusions Persistent pain symptoms were frequent in critical illness survivors and specialized management remained infrequent. Innovative approaches must be developed in the ICU to minimize the consequences of pain. Trial registration. NCT04817696. Registered March 26, 2021.
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Effect of the COVID-19 pandemic on obesity and it is risk factors: a systematic review - BMC Public HealthBackground Coronavirus disease (COVID-19) is a contagious disease caused by the severe acute new coronavirus called SARS-CoV-2. Devastating social, economic, and health service utilisation-related activities. Increased burden and lifestyle changes due to confinement. Objective This study aimed to investigate and determine the determinants of obesity during the coronavirus disease (COVID-19) pandemic from 2019 to 2023. Methods Observational studies published between December 2019 and January 2023 were thoroughly searched using a PRISMA flow chart. PubMed, Google Scholar, Web of Science, HINARI, Scopus, and Embase databases were used. Two reviewers independently identified and critically evaluated the relevant literature. Studies that reported weight gain or involved BMI measurements of 25 kg/m2 or BMI z-scores for children during the COVID-19 lockdown were selected for inclusion. The Newcastle–Ottawa Scale (NOS) was used as a quality assessment instrument in nonrandomised studies to evaluate study quality. All the contributing determinants of weight increase were identified, gathered, and synthesised. Results This systematic review identified 40 studies with a total population of 5,681,813 from 22 countries, of which 74.6% were male. The sample size from included articles ranged from 37 to 5,315,435. Of the 40 selected articles, 24 focused on adults, five on adolescents, three on children, and eight on children and adolescents. Physical inactivity, sedentary behaviour, bad eating habits, behavioural lifestyle, excessive stress, depression, anxiety, behavioural risk factors, sex, and ethnic minorities were associated with obesity during the COVID-19 pandemic lockdown. Conclusion During the COVID-19 pandemic, physical inactivity, sedentary lifestyle, and poor eating patterns were the most common risk factors for obesity. Additionally, unhealthy eating habits, excessive behavioural stress, depression, anxiety, low mood, age, gender, and ethnic minorities have been ident
Source: BioMedCentral - 🏆 22. / 71 Read more »