Within each of these 18 cells, raw ADOS totals were mapped onto a 10-point severity metric. After considering a variety of approaches, severity scores 1–3 were set so as to represent the distribution of raw scores receiving a nonspectrum ADOS classification within that calibration cell, severity scores 4–5 represented ASD-classification ADOS totals, and 6–10 represented raw totals receiving an autism classification within that cell. ADOS classification thresholds were determined by the revised algorithm relevant to each calibration cell. The range of raw totals corresponding to each point on the severity metric was determined by the percentiles of available data associated with each severity point within a classification range. Lower severity scores are associated with less autism impairment.
Cellular Senescence
This phenotype is triggered by various intrinsic and extrinsic factors, such as telomere attrition, DNA damage, oxidative stress, and oncogenic signaling.
Senescent cells exhibit distinct morphological and functional changes, including enlarged and flattened morphology, increased senescence-associated β-galactosidase activity, and the secretion of a pro-inflammatory senescence-associated secretory phenotype (SASP).
Cellular senescence plays a crucial role in organismal aging, tissue homeostasis, and the development of age-related diseases.
Understanfing the mechanisms and regulators of cellular senescence is crucial for developing targeted interventions to mitigate the detrimental effects of this process and promote healthy aging.
The study of cellular senescence has important implications for fields such as regenerative medicine, cancer biology, and age-related disease research.
Most cited protocols related to «Cellular Senescence»
To estimate “pure” epigenetic aging effects that are not influenced by differences in blood cell counts (“intrinsic” epigenetic age acceleration, IEAA), we obtained the residual resulting from a multivariate regression model of epigenetic age on chronological age and various blood immune cell counts (naive CD8+ T cells, exhausted CD8+ T cells, plasmablasts, CD4+ T cells, natural killer cells, monocytes, and granulocytes) imputed from methylation data.
Extrinsic epigenetic age acceleration measures capture both cell intrinsic methylation changes and extracellular changes in blood cell composition. Our measure of EEAA is defined using the following three steps. First, we calculated the epigenetic age measure from Hannum et al [2 (link)], which already correlated with certain blood cell types [5 (link)]. Second, we increased the contribution of immune blood cell types to the age estimate by forming a weighted average of Hannum's estimate with 3 cell types that are known to change with age: naïve (CD45RA+CCR7+) cytotoxic T cells, exhausted (CD28-CD45RA-) cytotoxic T cells, and plasmablasts using the Klemera-Doubal approach [32 (link)]. The weights used in the weighted average are determined by the correlation between the respective variable and chronological age [32 (link)]. The weights were chosen on the basis of the WHI data. Thus, the same (static) weights were used for all data sets. EEAA was defined as the residual variation resulting from a univariate model regressing the resulting age estimate on chronological age. By construction, EEAA is positively correlated with the estimated abundance of exhausted CD8+ T cells, plasmablast cells, and a negative correlated with naive CD8+ T cells. Blood cell counts were estimated based on DNA methylation data as described in the next section. By construction, the measures of EEAA track both age related changes in blood cell composition and intrinsic epigenetic changes. None of our four measures of epigenetic age acceleration are correlated with chronological age.
The first part of our work consisted in finding which genes driving CS are also associated with ARDs or with longevity, using the following data sources:
Human genes associated with CS: CellAge build 1.
Human genes associated with human aging: GenAge human build 19.
Human orthologues of model organisms’ genes associated with longevity: proOrthologuesPub.tsv and antiOrthologuesPub.tsv file (
Human oncogenes: Oncogene database (
Human tumor suppressor gene database: TSGene 2.0 (
Human genes associated with ARDs (
Human genes differentially expressed with age from the GTEx project (v7, January 2015 release) [32 (link), 43 (link)].
Most recents protocols related to «Cellular Senescence»
Example 8
Administration of bleomycin, a DNA damaging agent, to the anterior chamber of the mouse or rabbit eye leads to cellular senescence, as detected by the induction of p16 transcript in the trabecular meshwork.
To induce a senescent phenotype in the trabecular meshwork in vivo, C57Bl/6 mice (aged 8 to 10 weeks) were injected intracamerally with 2 μL of 0.0075 U bleomycin sulfate. In the rabbit, 30 μL of 0.0075 U bleomycin sulfate were injected intracamerally in New Zealand white rabbits. Eyes were enucleated 14 days post-bleomycin injury and TM-enriched samples were micro-dissected. To determine change in senescent cells, RNA was isolated from TM and qPCR analysis was done to assess the effect of bleomycin on p16 mRNA levels.
Example 1
A primary goal of the present invention was to develop yeast strains highly effective at metabolizing galactose. However, galactose-metabolizing yeasts are uncommon; yeasts typically prefer glucose, a carbohydrate source known to strongly suppress the expression of genes needed to metabolize other carbohydrates such as galactose (See Escalante-Chong et al., “Galactose metabolic genes in yeast respond to a ratio of galactose and glucose.” Proc Nat'l Acad Sci, USA 112:1636-41 (2015)). Wild type yeast strains may thus be prevented from utilizing any carbohydrates when glucose is present. Even if a yeast does have the capability to use other carbohydrate sources, it may occur late in the growth process, only after glucose has been completely depleted. Therefore, a particular type of galactose-metabolizing yeast was developed that degrades galactose in the presence of glucose. The present invention provides methods for adaptively evolving yeast according to this process.
To assess the ability of a yeast strain to degrade galactose, its growth was evaluated on media containing galactose in presence or absence of glucose. The following strains were tested: the commercially available strains Saccharomyces cerevisiae (N) (Natureland, Saccharomyces boulardii (SB) (Jarrow, Santa Fe Springs, CA), and Saccharomyces boulardii (B) (Biocodex, Redwood City, CA). Additional strains included in the screening were isolated from food containing large amounts of galactose such as dairy products and legumes stored at room temperature for over two weeks.
Cultures of various strains were initiated from a single colony on agar plates or from glycerol stocks, and grown in liquid YP medium (1% yeast extract, 2% peptone; Teknova) by incubation at 30° C. with agitation at 125 rpm (Murakami & Kaeberlein “Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells.” J Visual Exp (27) (2009)). Overnight yeast cultures initiated in duplicate in liquid YP medium were used as pre-cultures to initiate growth efficiency experiments in liquid CM (Synthetic Complete Minimal Medium, 0.5% Ammonium Sulfate, Teknova) containing 2% galactose alone as the sole carbon source, 2% glucose alone as the sole carbon source, or galactose and glucose. Culture growth of cultures set at 30° C. under static conditions was monitored over time by measuring optical density (OD) at 600 nm (OD600) using a spectrophotometer.
Growth—was evaluated for several strains. As illustrated in Table 1, one of the evolved clone exhibited the lowest doubling time, which remained at the same level independently of the carbohydrate source and growth conditions.
Example 6
The efficacy of model compound UBX1967 was studied in the mouse oxygen-induced retinopathy (OIR) model, which provides an in vivo model of retinopathy of prematurity (ROP) and diabetic retinopathy.
C57Bl/6 mouse pups and their CD1 foster mothers were exposed to a high oxygen environment (75% 02) from postnatal day 7 (P7) to P12. At P12, animals were injected intravitreally with 1 μl test compound (200, 20, or 2 uM) formulated in 1% DMSO, 10% Tween-80, 20% PEG-400, and returned to room air until P17. Eyes were enucleated at P17 and retinas dissected for either vascular staining or qRT-PCR. To determine avascular or neovascular area, retinas were flatmounted, and stained with isolectin B4 (IB4) diluted 1:100 in 1 mM CaCl2. For quantitative measurement of senesecence markers (e.g., Cdkn2a, Cdkn1a, 116, Vegfa), qPCR was performed. RNA was isolated and cDNA was generated by reverse-transcription, which was used for qRT-PCR of the selected transcripts.
These results show that a single ocular injection of UBX1967 can functionally inhibit pathogenic angiogenesis and promote vascular repair in this key OIR disease model. We believe that efficacy of UBX1967 in the OIR model is due to elimination of senescent cells and accompanying SASP that propagates senescence in retinal cells and promotes neovascularization of retinal vessels.
The prognostic significance of cellular senescence-related lncRNAs was initially determined using univariate Cox regression. Least absolute shrinkage and selection operator (LASSO) regression was used to integrate the cellular senescence-related lncRNAs with p < 0.05 in univariate analysis. The LASSO results were then included in a multivariate Cox model to generate a risk score. A risk score was calculated using a linear combination of cellular senescence-related lncRNA expression levels multiplied by a regression coefficient (β): risk score = (expression of lncRNAi). Based on the median risk score, the patients were categorized into high-risk and low-risk groups. The log-rank test was used to compare the survival differences between the two groups.
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More about "Cellular Senescence"
This phenotype is triggered by various intrinsic and extrinsic factors, such as telomere attrition, DNA damage, oxidative stress, and oncogenic signaling.
Senescent cells exhibit distinct morphological and functional changes, including enlarged and flattened morphology, increased senescence-associated β-galactosidase (SA-β-gal) activity, and the secretion of a pro-inflammatory senescence-associated secretory phenotype (SASP).
Cellular senescence plays a crucial role in organismal aging, tissue homeostasis, and the development of age-related diseases.
Understanding the mechanisms and regulators of cellular senescence is crucial for developing targeted interventions to mitigate the detrimental effects of this process and promote healthy aging.
The study of cellular senescence has important implications for fields such as regenerative medicine, cancer biology, and age-related disease research.
Researchers can leverage various tools and assays to study cellular senescence, including the Senescence β-Galactosidase Staining Kit, SA-β-gal staining kit, Senescence Cells Histochemical Staining Kit, Senescence Detection Kit, and Cellular Senescence Assay Kit.
These kits often utilize the increased activity of β-galactosidase, a hallmark of senescent cells, to detect and quantify senescent cell populations.
Additionally, fetal bovine serum (FBS) can be used in cell culture to support the growth and maintenance of senescent cells.
By incorporating these insights and related terms, researchers can optimize their cellular senescence research and uncover new strategies for promoting healthy aging and mitigating age-related diseases.