scholarly journals Haskap Berry Phenolic Subclasses Differentially Impact Cellular Stress Sensing in Primary and Immortalized Dermal Fibroblasts

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2643
Author(s):  
Lily R. Zehfus ◽  
Zoe E. Gillespie ◽  
Carla Almendáriz-Palacios ◽  
Nicholas H. Low ◽  
Christopher H. Eskiw
Keyword(s):  
Free Radicals ◽  
Mammalian Target Of Rapamycin ◽  
Nuclear Factor Κb ◽  
Dermal Fibroblasts ◽  
Sirtuin 1 ◽  
Population Doubling ◽  
Published Data ◽  
Stress Sensing ◽  
Berry Extracts ◽  
Doubling Times

It is generally accepted that dietary phenolics from fruits are of significant importance to human health. Unfortunately, there is minimal published data on how differences in phenolic structure(s) impact biological pathways at cellular and molecular levels. We observed that haskap berry extracts isolated with ethanol:formic acid:water or phenolic subclass fractions separated using different concentrations of ethanol (40% and 100%) impacted cell growth in a positive manner. All fractions and extracts significantly increased population doubling times. All extracts and fractions reduced intracellular free radicals; however, there were differences in these effects, indicating different abilities to scavenge free radicals. The extracts and fractions also exhibited differing impacts on transcripts encoding the antioxidant enzymes (CAT, SOD1, GPX1, GSS and HMOX1) and the phosphorylation state of nuclear factor-κB (NF-κB). We further observed that extracts and fractions containing different phenolic structures had divergent impacts on the mammalian target of rapamycin (mTOR) and sirtuin 1 (SIRT1). siRNA-mediated knockdown of SIRT1 transcripts demonstrated that this enzyme is key to eliciting haskap phenolic(s) impact on cells. We postulate that phenolic synergism is of significant importance when evaluating their dietary impact.

Culture of Arterial Endothelial Cells

1975 ◽  
Vol 34 (03) ◽  
pp. 825-839 ◽  
Author(s):  
Francois M Booyse ◽  
Bonnie J Sedlak ◽  
Max E Rafelson
Keyword(s):  
Endothelial Cells ◽  
Calf Serum ◽  
Intercellular Junctions ◽  
Cell Number ◽  
Population Doubling ◽  
Homogeneous Population ◽  
Bovine Endothelial Cells ◽  
Pinocytotic Vesicles ◽  
Arterial Endothelial Cells ◽  
Doubling Times

SummaryArterial endothelial cells were obtained from bovine aortae by mild treatment with collagenase and medium perfusion. These cells were cultured in RPMI-1640 medium containing 15 mM Hepes buffer and 35% fetal calf serum at pH 7.35. Essentially ah (90–95%) the effluent cells were viable and 80% of these cells attached to the substratum within 1 hour. Small patches of attached cells coalesced to form confluent monolayers in 3–5 days. Confluent monolayers of endothelial cells consisted of a homogeneous population of tightly packed, polygonal cells. Selected cultures were serially subcultured (trypsin-EDTA) for 12–14 months (30–35 passages) without any apparent change in morphology or loss of growth characteristics. Primary and three-month old (15 passages) cultures had population doubling times of 32–34 hours and 29–31 hours, respectively. These cells (primary and subcultures) did not require a minimum cell number to become established in culture. Bovine endothelial cells (primary, first, fifth and thirteenth passages) were characterized ultrastructurally by the presence of Weibel-Palade bodies, pinocytotic vesicles and microfilaments and immunologically by the presence of thrombosthenin-like contractile proteins and Factor VIII antigen. The intercellular junctions of post-confluent cultures stained specifically with silver nitrate. From these data, we concluded that identifiable endothelial cells could be obtained from bovine aortae and cultured and maintained for prolonged periods of time.

mTOR inhibition as a possible pharmacological target in the management of systemic inflammatory response and associated neuroinflammation by lipopolysaccharide challenge in rats

2021 ◽  
Author(s):  
Demet Sinem Guden ◽  
Meryem Temiz-Resitoglu ◽  
Sefika Pınar Senol ◽  
Deniz Kibar ◽  
Sakir Necat Yilmaz ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Microglial Activation ◽  
Critical Role ◽  
Mammalian Target Of Rapamycin ◽  
Hypoxia Inducible Factor ◽  
B Cell Lymphoma ◽  
Nuclear Factor Κb ◽  
Systemic Inflammatory Response ◽  
Mtor Inhibition ◽  
Factor Α

Neuroinflammation plays a critical role during sepsis triggered by microglial activation. Mammalian target of rapamycin (mTOR) has gained attraction in neuroinflammation, however, the mechanism remains unclear. Our goal was to assess the effects of mTOR inhibition by rapamycin on inflammation, microglial activation, oxidative stress, and apoptosis associated with the changes in the inhibitor-κB (IκB)-α/nuclear factor-κB (NF-κB)/hypoxia-inducible factor-1α (HIF-1α) pathway activity following a systemic challenge with lipopolysaccharide (LPS). Rats received saline (10 ml/kg), LPS (10 mg/kg), and/or rapamycin (1 mg/kg) via intraperitoneally. Inhibition of mTOR by rapamycin blocked phosphorylated form of ribosomal protein S6, NF-κB p65 activity by increasing degradation of IκB-α in parallel with HIF-1α expression increased by LPS in the kidney, heart, lung, and brain tissues. Rapamycin attenuated the increment in the expression of tumor necrosis factor-α and interleukin-1β, the inducible nitric oxide synthase, gp91<sup>phox</sup>, and p47<sup>phox</sup> in addition to nitrite levels elicited by LPS in tissues or sera. Concomitantly, rapamycin treatment reduced microglial activation, brain expression of caspase-3, and Bcl-2-associated X protein while increased expression of B-cell lymphoma 2 induced by LPS. Overall, this study supports the hypothesis that mTOR contributes to the detrimental effect of LPS-induced systemic inflammatory response associated with neuroinflammation via IκB-α/NF-κB/HIF-1α signaling pathway.

scholarly journals Angiogenic Deficiency and Adipose Tissue Dysfunction Are Associated with Macrophage Malfunction in SIRT1−/− Mice

Endocrinology ◽  
2012 ◽  
Vol 153 (4) ◽  
pp. 1706-1716 ◽  
Author(s):  
Fen Xu ◽  
David Burk ◽  
Zhanguo Gao ◽  
Jun Yin ◽  
Xia Zhang ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Adipocyte Differentiation ◽  
Nuclear Factor Κb ◽  
The Body ◽  
Sirtuin 1 ◽  
Vascular Density ◽  
Peroxisome Proliferator ◽  
Wild Type ◽  
Peroxisome Proliferator Activated Receptor

The histone deacetylase sirtuin 1 (SIRT1) inhibits adipocyte differentiation and suppresses inflammation by targeting the transcription factors peroxisome proliferator-activated receptor γ and nuclear factor κB. Although this suggests that adiposity and inflammation should be enhanced when SIRT1 activity is inactivated in the body, this hypothesis has not been tested in SIRT1 null (SIRT1−/−) mice. In this study, we addressed this issue by investigating the adipose tissue in SIRT1−/− mice. Compared with their wild-type littermates, SIRT1 null mice exhibited a significant reduction in body weight. In adipose tissue, the average size of adipocytes was smaller, the content of extracellular matrix was lower, adiponectin and leptin were expressed at 60% of normal level, and adipocyte differentiation was reduced. All of these changes were observed with a 50% reduction in capillary density that was determined using a three-dimensional imaging technique. Except for vascular endothelial growth factor, the expression of several angiogenic factors (Pdgf, Hgf, endothelin, apelin, and Tgf-β) was reduced by about 50%. Macrophage infiltration and inflammatory cytokine expression were 70% less in the adipose tissue of null mice and macrophage differentiation was significantly inhibited in SIRT1−/− mouse embryonic fibroblasts in vitro. In wild-type mice, macrophage deletion led to a reduction in vascular density. These data suggest that SIRT1 controls adipose tissue function through regulation of angiogenesis, whose deficiency is associated with macrophage malfunction in SIRT1−/− mice. The study supports the concept that inflammation regulates angiogenesis in the adipose tissue.

scholarly journals A review of the life cycles and life-history adaptations of pelagic tunicates to environmental conditions

2011 ◽  
Vol 69 (3) ◽  
pp. 358-369 ◽  
Author(s):  
Don Deibel ◽  
Ben Lowen
Keyword(s):  
Life History ◽  
Environmental Conditions ◽  
Generation Time ◽  
Egg Production ◽  
Food Concentration ◽  
Life Cycles ◽  
Population Doubling ◽  
Event Scale ◽  
Generation Times ◽  
Doubling Times

Abstract Deibel, D., and Lowen, B. 2012. A review of the life cycles and life-history adaptations of pelagic tunicates to environmental conditions. – ICES Journal of Marine Science, 69: 358–369. Phylogeny, life cycles, and life-history adaptations of pelagic tunicates to temperature and food concentration are reviewed. Using literature data on lifetime egg production and generation time of appendicularians, salps, and doliolids, rmax, the maximum rate of lifetime reproductive fitness, is calculated as a common metric of adaptation to environmental conditions. The rmax values are high for all three groups, ranging from ∼0.1 to 1.9 d−1, so population doubling times range from ∼8 h to 1 week. These high values of rmax are attributable primarily to short generation times, ranging from 2 to 50 d. Clearly, pelagic tunicates are adapted to event-scale (i.e. days to weeks) rather than seasonal-scale changes in environmental conditions. Although they are not closely related phylogenetically, all three groups have a unique life-history adaptation promoting high lifetime fitness. Appendicularians have late oocyte selection, salps are viviparous, and doliolids possess a polymorphic asexual phase. There has been little research on hermaphroditic appendicularians, on large oceanic salps, and on doliolids generally. Research is needed on factors regulating generation time, on the heritability of life-history traits, and on age- and size-specific rates of mortality.

scholarly journals Sirtuin 1 suppresses nuclear factor κB induced transactivation and pro-inflammatory cytokine expression in cat fibroblast cells

2015 ◽  
Vol 77 (12) ◽  
pp. 1681-1684 ◽  
Author(s):  
Shingo ISHIKAWA ◽  
Hiroshi TAKEMITSU ◽  
Makoto HABARA ◽  
Nobuko MORI ◽  
Ichiro YAMAMOTO ◽  
...  
Keyword(s):  
Inflammatory Cytokine ◽  
Nuclear Factor Κb ◽  
Cytokine Expression ◽  
Sirtuin 1 ◽  
Fibroblast Cells ◽  
Nuclear Factor

The Effect of Follicular Fluid on the Proliferation and Osteoblastic Differentiation of Human Bone Marrow Mesenchymal Stem Cells

2019 ◽  
Author(s):  
Atefeh Soltani ◽  
Saeid Abroun ◽  
Mojtaba Rezazadeh Valojerdi ◽  
Bahareh Vahidianfar ◽  
Elahe Sadat Hosseini
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Cell Culture ◽  
Bone Marrow ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Population Doubling ◽  
Growth Factor Beta ◽  
Doubling Times

Background and Aims: Bone marrow-derived mesenchymal stem cells (BM-MSCs) are a well-known source of multipotent adult stem cells. Despite using different methodologies of MSCs preparing for clinical applications, the top safest procedure to manipulate these cells, has not yet been determined. Recently, ex-vivo expansion of MSCs for their subsequent implantation, using some biological product, is suggested instead of fetal bovine serum (FBS). Previous studies have shown the effect of follicular fluid (FF) (a dynamic fluid in ovarian follicle) as an additive component in cell culture. Hence, this study aimed to decipher its role on the human BM-MSC proliferation.Materials and Methods: In this study, BM-MSCs at 3rd passage were cultivated in the presence of 20% FF (group I), 10% FF+ FBS 10% (group II) and FBS 20% as control group. The capacity of proliferation as calculating population doubling times and gene expression levels of stem cell factor, stromal cell-derived factor 1, and transforming growth factor beta were analyzed in osteogeneic media to examine the impacts of FF on osteogenesis of MSCs.Results: Our results corroborated an up-regulatory effect of FF on the proliferation of BM-MSCs by shorter population doubling times in the group II of treated cells and an increase in gene expression level of osteocalcin and transforming growth factor beta in the presence of higher concentrations of FF in cell culture  FF 20% and 10%, respectively.Conclusions: FF is a potent mitogen for cell proliferation. FF may be an efficient substitution of FBS in ex-vivo cell culture, eliminating zoonotic infections and immunological reactions.

Mathematical Determination of Cell Population Doubling Times for Multiple Cell Lines

2012 ◽  
Vol 74 (10) ◽  
pp. 2510-2534 ◽  
Author(s):  
Liene Daukste ◽  
Britta Basse ◽  
Bruce C. Baguley ◽  
David J. N. Wall
Keyword(s):  
Cell Lines ◽  
Cell Population ◽  
Population Doubling ◽  
Multiple Cell ◽  
Doubling Times

scholarly journals Growth factor regulation of hyaluronan synthesis and degradation in human dermal fibroblasts: importance of hyaluronan for the mitogenic response of PDGF-BB

2007 ◽  
Vol 404 (2) ◽  
pp. 327-336 ◽  
Author(s):  
Lingli Li ◽  
Trias Asteriou ◽  
Berit Bernert ◽  
Carl-Henrik Heldin ◽  
Paraskevi Heldin
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Fold Increase ◽  
Nuclear Factor Κb ◽  
Transforming Growth Factor Β1 ◽  
Dermal Fibroblasts ◽  
Mitogen Activated Protein Kinase ◽  
Signalling Pathways ◽  
Human Dermal Fibroblasts ◽  
High Level

The glycosaminoglycan hyaluronan is important in many tissuerepair processes. We have investigated the synthesis of hyaluronan in a panel of cell lines of fibroblastic and epithelial origin in response to PDGF (platelet-derived growth factor)-BB and other growth factors. Human dermal fibroblasts exhibited the highest hyaluronan-synthesizing activity in response to PDGF-BB. Analysis of HAS (hyaluronan synthase) and HYAL (hyaluronidase) mRNA expression showed that PDGF-BB treatment induced a 3-fold increase in the already high level of HAS2 mRNA, and increases in HAS1 and HYAL1 mRNA, whereas the levels of HAS3 and HYAL2 mRNA were not affected. Furthermore, PDGF-BB also increased the amount and activity of HAS2 protein, but not of HYAL1 and HYAL2 proteins. Using inhibitors for MEK1/2 [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase 1/2] (U0126) and for PI3K (phosphoinositide 3-kinase) (LY294002), as well as the SN50 inhibitor, which prevents translocation of the active NF-κB (nuclear factor κB) to the nucleus, we observed a complete inhibition of both HAS2 transcriptional activity and hyaluronan synthesis, whereas inhibitors of other signalling pathways were without any significant effect. TGF-β1 (transforming growth factor-β1) did not increase the activity of hyaluronan synthesis in dermal fibroblasts, but increased the activity of HYALs. Importantly, inhibition of hyaluronan binding to its receptor CD44 by the monoclonal antibody Hermes-1, inhibited PDGF-BB-stimulated [3H]thymidine incorporation of dermal fibroblasts. We conclude that the ERK MAPK and PI3K signalling pathways are necessary for the regulation of hyaluronan synthesis by PDGF-BB, and that prevention of its binding to CD44 inhibits PDGF-BB-induced cell growth.

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