scholarly journals Safety Profile of Caesalpinia spinosa Aqueous Extract Tested in Oreochromis niloticus Toward Its Application in Dermocosmetics

2021 ◽  
Vol 2 ◽  
Author(s):  
David Salirrosas ◽  
Nataly Reategui-Pinedo ◽  
Jan Pier Crespo ◽  
Linda Sánchez-Tuesta ◽  
Mónica Arqueros ◽  
...  
Keyword(s):  
Dna Damage ◽  
Aqueous Extract ◽  
Oreochromis Niloticus ◽  
Ex Vivo ◽  
Pharmaceutical Products ◽  
Cell Permeability ◽  
Internal Validation ◽  
Juvenile Stages ◽  
Morphological Deformities

Herbal extracts have been widely used in dermocosmetics as a source of biomolecules and also as a natural claim. Fruits from Caesalpinia spinosa show great potential for their polyphenolic content, preservative, and film-forming features, as previously reported in specialized literature; however, the toxicity requires investigation. We explored Oreochromis niloticus (tilapia) in larval, alevins, and juvenile stages to evaluate the ex vivo and in vivo genotoxicity and in vivo acute and chronic toxicity of C. spinosa aqueous extract in different concentrations. Cytotoxicity, animal behavior, morphological deformities, and DNA damage were evaluated. Our results showed genotoxic effect in ex vivo tests, but no DNA damage in in vivo erythrocytes. We suggest a mechanism of cell permeability involved in the toxicity of C. spinosa aqueous extract. Internal validation showed the feasibility of O. niloticus applied for toxicity evaluation. Further studies could contribute for better understanding the uses and safety of C. spinosa in cosmetics and topical pharmaceutical products.

scholarly journals Extra-gastric expression of the proton pump H+/K+-ATPase in the gills and kidney of the teleost Oreochromis niloticus

2020 ◽  
Vol 223 (16) ◽  
pp. jeb214890
Author(s):  
Ebtesam Ali Barnawi ◽  
Justine E. Doherty ◽  
Patrícia Gomes Ferreira ◽  
Jonathan M. Wilson
Keyword(s):  
Oreochromis Niloticus ◽  
Cellular Localization ◽  
Ex Vivo ◽  
Uptake Inhibition ◽  
Custom Made ◽  
Potassium Regulation ◽  
Immunohistochemical Techniques ◽  
Collecting Tubules ◽  
First Time

ABSTRACTPotassium regulation is essential for the proper functioning of excitable tissues in vertebrates. The H+/K+-ATPase (HKA), which is composed of the HKα1 (gene: atp4a) and HKβ (gene: atp4b) subunits, has an established role in potassium and acid–base regulation in mammals and is well known for its role in gastric acidification. However, the role of HKA in extra-gastric organs such as the gill and kidney is less clear, especially in fishes. In the present study in Nile tilapia, Oreochromis niloticus, uptake of the K+ surrogate flux marker rubidium (Rb+) was demonstrated in vivo; however, this uptake was not inhibited with omeprazole, a potent inhibitor of the gastric HKA. This contrasts with gill and kidney ex vivo preparations, where tissue Rb+ uptake was significantly inhibited by omeprazole and SCH28080, another gastric HKA inhibitor. The cellular localization of this pump in both the gill and kidney was demonstrated using immunohistochemical techniques with custom-made antibodies specific for Atp4a and Atp4b. Antibodies against the two subunits showed the same apical ionocyte distribution pattern in the gill and collecting tubules/ducts in the kidney. Atp4a antibody specificity was confirmed by western blotting. RT-PCT was used to confirm the expression of both subunits in the gill and kidney. Taken together, these results indicate for the first time K+ (Rb+) uptake in O. niloticus and that HKA is implicated, as shown through the ex vivo uptake inhibition by omeprazole and SCH28080, verifying a role for HKA in K+ absorption in the gill's ionocytes and collecting tubule/duct segments of the kidney.

scholarly journals Osmanthus fragrans Flower Aqueous Extract and its Enriched Acteoside inhibit Melanogenesis and Ultraviolet-induced Pigmentation

2018 ◽  
Vol 13 (5) ◽  
pp. 1934578X1801300
Author(s):  
Shuo Liu ◽  
Zhen Zhao ◽  
Zhijun Huo ◽  
Zhiru Xu ◽  
Yan Zhong ◽  
...  
Keyword(s):  
Aqueous Extract ◽  
Ex Vivo ◽  
Melanoma Cells ◽  
B16 Melanoma ◽  
Tyrosinase Activity ◽  
In Vivo Model ◽  
Mrna Levels ◽  
Osmanthus Fragrans ◽  
B16 Melanoma Cells

The Osmanthus fragrans flower (OFF) is commonly used as an additive for tea in China and as a traditional medicine to treat dysentery, asthma and hepatitis. In the current study, we have acquired the aqueous extract of the dried OFF (OFFE) and determined its enriched acteoside contents. However, whether OFFE and acteoside can modulate melanogenesis and pigmentation has yet to be determined. We here provide novel data revealing that OFFE and acteoside inhibit melanogenesis induced by α-MSH in B16 melanoma cells via the MITF-tyrosinase signaling pathway. Treatment with α-MSH (1μM) enhanced melanin levels and tyrosinase activity, up-regulated the mRNA levels of MITF and tyrosinase and increased the dendritic number in B16 melanoma cells, effects all being intervened by OFFE and acteoside. Of interest, OFFE and acteoside showed no direct inhibition of tyrosinase activity as revealed by our ex vivo tyrosinase activity assay. In addition, OFFE produced a depigmenting action on UVB-induced hyperpigmentation in guinea pigs, as shown by the improved skin brightness and the decreased melanin staining. Our data have demonstrated that OFFE can alter melanogenesis via modulating the MITF-tyrosinase signaling thereby leading to its depigmenting action in the in vivo model. OFFE could be a substitute for acteoside as a promising skin-whitening agent.

Apoptosis and DNA damage in type 2 alveolar epithelial cells cultured from hyperoxic rats

1998 ◽  
Vol 274 (5) ◽  
pp. L714-L720 ◽  
Author(s):  
Sue Buckley ◽  
Lora Barsky ◽  
Barbara Driscoll ◽  
Kenneth Weinberg ◽  
Kathryn D. Anderson ◽  
...  
Keyword(s):  
Dna Damage ◽  
Epithelial Cells ◽  
Cellular Response ◽  
Ex Vivo ◽  
Keratinocyte Growth Factor ◽  
Alveolar Epithelial Cells ◽  
Alveolar Epithelial ◽  
Dna Laddering

Apoptosis is a genetically controlled cellular response to developmental stimuli and environmental insult that culminates in cell death. Sublethal hyperoxic injury in rodents is characterized by a complex but reproducible pattern of lung injury and repair during which the alveolar surface is damaged, denuded, and finally repopulated by type 2 alveolar epithelial cells (AEC2). Postulating that apoptosis might occur in AEC2 after hyperoxic injury, we looked for the hallmarks of apoptosis in AEC2 from hyperoxic rats. A pattern of increased DNA end labeling, DNA laddering, and induction of p53, p21, and Bax proteins, strongly suggestive of apoptosis, was seen in AEC2 cultured from hyperoxic rats when compared with control AEC2. In contrast, significant apoptosis was not detected in freshly isolated AEC2 from oxygen-treated rats. Thus the basal culture conditions appeared to be insufficient to ensure the ex vivo survival of AEC2 damaged in vivo. The oxygen-induced DNA strand breaks were blocked by the addition of 20 ng/ml of keratinocyte growth factor (KGF) to the culture medium from the time of plating and were partly inhibited by Matrigel or a soluble extract of Matrigel. KGF treatment resulted in a partial reduction in the expression of the p21, p53, and Bax proteins but had no effect on DNA laddering. We conclude that sublethal doses of oxygen in vivo cause damage to AEC2, resulting in apoptosis in ex vivo culture, and that KGF can reduce the oxygen-induced DNA damage. We speculate that KGF plays a role as a survival factor in AEC2 by limiting apoptosis in the lung after acute hyperoxic injury.

scholarly journals The Injury Response to DNA Damage Promotes Anti-Tumor Immunity

2020 ◽  
Author(s):  
Ganapathy Sriram ◽  
Lauren Milling ◽  
Jung-Kuei Chen ◽  
Wuhbet Abraham ◽  
Erika D. Handly ◽  
...  
Keyword(s):  
Dna Damage ◽  
T Cells ◽  
Dendritic Cells ◽  
T Cell ◽  
Tumor Cells ◽  
Immune Checkpoint ◽  
Ex Vivo ◽  
Injured Cell ◽  
Ifn Γ

ABSTRACTInhibition of immune checkpoints has shown promising results in the treatment of certain tumor types. However, the majority of cancers do not respond to immune checkpoint inhibition (ICI) treatment, indicating the need to identify additional modalities that enhance the response to immune checkpoint blockade. In this study, we identified a tumor-tailored approach using ex-vivo DNA damaging chemotherapy-treated tumor cells as a live injured cell adjuvant. Using an optimized ex vivo system for dendritic cell-mediated T-cell IFN-γ induction in response to DNA-damaged tumor cells, we identified specific dose-dependent treatments with etoposide and mitoxantrone that markedly enhance IFN-γ production by T-cells. Unexpectedly, the immune-enhancing effects of DNA damage failed to correlate with known markers of immunogenic cell death or with the extent of apoptosis or necroptosis. Furthermore, dead tumor cells alone were not sufficient to promote DC cross-presentation and induce IFN-γ in T-cells. Instead, the enhanced immunogenicity resided in the fraction of injured cells that remained alive, and required signaling through the RIPK1, NF-kB and p38MAPK pathways. Direct in vivo translation of these findings was accomplished by intra-tumoral injection of ex vivo etoposide-treated tumor cells as an injured cell adjuvant, in combination with systemic anti-PD1/CTLA4 antibodies. This resulted in increased intra-tumoral CD103+ dendritic cells and circulating tumor antigen-specific CD8+ T-cells, leading to enhanced anti-tumor immune responses and improved survival. The effect was abrogated in BATF3-deficient mice indicating that BATF3+ DCs are required for appropriate T-cell stimulation by live but injured DNA-damaged tumor cells. Notably, injection of the free DNA-damaging drug directly into the tumor failed to elicit such an enhanced anti-tumor response as a consequence of simultaneous damage to dendritic cells and T-cells. Finally, the DNA damage induced injured cell adjuvant and systemic ICI combination, but not ICI alone, induced complete tumor regression in a subset of mice who were then able to reject tumor re-challenge, indicating induction of a long-lasting anti-tumor immunological memory by the injured cell adjuvant treatment in vivo.

scholarly journals PTEN mutant NSCLC require ATM to suppress pro-apoptotic signalling and evade radiotherapy

2021 ◽  
Author(s):  
Thomas Fischer ◽  
Oliver Hartmann ◽  
Michaela Reissland ◽  
Cristian Prieto-Garcia ◽  
Kevin Klann ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Dna Damage ◽  
Western Blot ◽  
Colony Formation ◽  
Ex Vivo ◽  
Radiation Sensitivity ◽  
Genetic Alterations ◽  
Altered Expression ◽  
The Impact

Background: Despite advances in treatment of patients with non-small cell lung cancer, carriers of certain genetic alterations are prone to failure. One such factor frequently mutated, is the tumor suppressor PTEN. These tumors are supposed to be more resistant to radiation, chemo- and immunotherapy. Methods: Using CRISPR genome editing, we deleted PTEN in a human tracheal stem cell-like cell line as well generated primary murine NSCLC, proficient or deficient for Pten, in vivo. These models were used to verify the impact of PTEN loss in vitro and in vivo by immunohistochemical staining, western blot and RNA-Sequencing. Radiation sensitivity was assessed by colony formation and growth assays. To elucidate putative treatment options, identified via the molecular characterisation, PTEN pro- and deficient cells were treated with PI3K/mTOR/DNA-PK-inhibitor PI-103 or the ATM-inhibitors KU-60019 und AZD 1390. Changes in radiation sensitivity were assessed by colony-formation assay, FACS, western-blot, phospho-proteomic mass spectrometry and ex vivo lung slice cultures. Results: We demonstrate that loss of PTEN led to altered expression of transcriptional programs which directly regulate therapy resistance, resulting in establishment of radiation resistance. While PTEN-deficient tumor cells were not dependent on DNA PK for IR resistance nor activated ATR during IR, they showed a significant dependence for the DNA damage kinase ATM. Pharmacologic inhibition of ATM, via KU-60019 and AZD1390 at non-toxic doses, restored and even synergized with IR in PTEN-deficient human and murine NSCLC cells as well in a multicellular organotypic ex vivo tumor model. Conclusion: PTEN tumors are addicted to ATM to detect and repair radiation induced DNA damage. This creates an exploitable bottleneck. At least in cellulo and ex vivo we show that low concentration of ATM inhibitor is able to synergise with IR to treat PTEN-deficient tumors in genetically well-defined IR resistant lung cancer models.

scholarly journals Effects of wound dressings containing silver on skin and immune cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Kristina Nešporová ◽  
Vojtěch Pavlík ◽  
Barbora Šafránková ◽  
Hana Vágnerová ◽  
Pavel Odráška ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ex Vivo ◽  
Wound Dressings ◽  
In Vitro Toxicity ◽  
Skin Cells ◽  
Antimicrobial Efficiency ◽  
Activated Neutrophils ◽  
Stress Related Genes

Abstract Wound dressings with silver have been shown to be cytotoxic in vitro. However, the extrapolation of this cytotoxicity to clinical settings is unclear. We applied dressings with various forms of silver on porcine skin ex vivo and investigated silver penetration and DNA damage. We assessed antimicrobial efficacy, cytotoxicity to skin cells, and immune response induced by the dressings. All dressings elevated the DNA damage marker γ-H2AX and the expression of stress-related genes in explanted skin relative to control. This corresponded with the amount of silver in the skin. The dressings reduced viability, induced oxidative stress and DNA damage in skin cells, and induced the production of pro-inflammatory IL-6 by monocytes. The oxidative burst and viability of activated neutrophils decreased. The amount of silver released into the culture medium varied among the dressings and correlated with in vitro toxicity. However, antimicrobial efficiencies did not correlate strongly with the amount of silver released from the dressings. Antimicrobial efficiency and toxicity are driven by the form of silver and the construction of dressings and not only by the silver concentration. The damaging effects of silver dressings in ex vivo skin highlight the importance of thorough in vivo investigation of silver dressing toxicity.

scholarly journals Tissue-infiltrating macrophages mediate an exosome-based metabolic reprogramming upon DNA damage

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Evi Goulielmaki ◽  
Anna Ioannidou ◽  
Maria Tsekrekou ◽  
Kalliopi Stratigi ◽  
Ioanna K. Poutakidou ◽  
...  
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Glucose Transporter ◽  
Ex Vivo ◽  
Disease Onset ◽  
Metabolic Reprogramming ◽  
Inflammatory Stimuli ◽  
Underlying Mechanisms ◽  
Repair Defect

AbstractDNA damage and metabolic disorders are intimately linked with premature disease onset but the underlying mechanisms remain poorly understood. Here, we show that persistent DNA damage accumulation in tissue-infiltrating macrophages carrying an ERCC1-XPF DNA repair defect (Er1F/−) triggers Golgi dispersal, dilation of endoplasmic reticulum, autophagy and exosome biogenesis leading to the secretion of extracellular vesicles (EVs) in vivo and ex vivo. Macrophage-derived EVs accumulate in Er1F/− animal sera and are secreted in macrophage media after DNA damage. The Er1F/− EV cargo is taken up by recipient cells leading to an increase in insulin-independent glucose transporter levels, enhanced cellular glucose uptake, higher cellular oxygen consumption rate and greater tolerance to glucose challenge in mice. We find that high glucose in EV-targeted cells triggers pro-inflammatory stimuli via mTOR activation. This, in turn, establishes chronic inflammation and tissue pathology in mice with important ramifications for DNA repair-deficient, progeroid syndromes and aging.

scholarly journals A Critical Analysis of the AvailableIn VitroandEx VivoMethods to Study Retinal Angiogenesis

2017 ◽  
Vol 2017 ◽  
pp. 1-19 ◽  
Author(s):  
A. F. Moleiro ◽  
G. Conceição ◽  
A. F. Leite-Moreira ◽  
A. Rocha-Sousa
Keyword(s):  
Ex Vivo ◽  
Tube Formation ◽  
Cell Permeability ◽  
Retinal Diseases ◽  
Advantages And Disadvantages ◽  
Retinal Angiogenesis ◽  
Ideal Method ◽  
The Ideal

Angiogenesis is a biological process with a central role in retinal diseases. The choice of the ideal method to study angiogenesis, particularly in the retina, remains a problem. Angiogenesis can be assessed throughin vitroandin vivostudies. In spite of inherent limitations,in vitrostudies are faster, easier to perform and quantify, and typically less expensive and allow the study of isolated angiogenesis steps. We performed a systematic review of PubMed searching for original articles that appliedin vitroor ex vivo angiogenic retinal assays until May 2017, presenting the available assays and discussing their applicability, advantages, and disadvantages. Most of the studies evaluated migration, proliferation, and tube formation of endothelial cells in response to inhibitory or stimulatory compounds. Other aspects of angiogenesis were studied by assessing cell permeability, adhesion, or apoptosis, as well as by implementing organotypic models of the retina. Emphasis is placed on how the methods are applied and how they can contribute to retinal angiogenesis comprehension. We also discuss how to choose the best cell culture to implement these methods. When applied together,in vitroand ex vivo studies constitute a powerful tool to improve retinal angiogenesis knowledge. This review provides support for researchers to better select the most suitable protocols in this field.

scholarly journals DNA damage after long-term repetitive hyperbaric oxygen exposure

2009 ◽  
Vol 106 (1) ◽  
pp. 311-315 ◽  
Author(s):  
Michael Gröger ◽  
Sükrü Öter ◽  
Vladislava Simkova ◽  
Markus Bolten ◽  
Andreas Koch ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Endurance Training ◽  
Hyperbaric Oxygen ◽  
Oxidative Dna Damage ◽  
Ex Vivo ◽  
Pure Oxygen ◽  
Repetitive Exposure

A single exposure to hyperbaric oxygen (HBO), i.e., pure oxygen breathing at supra-atmospheric pressures, causes oxidative DNA damage in humans in vivo as well as in isolated lymphocytes of human volunteers. These DNA lesions, however, are rapidly repaired, and an adaptive protection is triggered against further oxidative stress caused by HBO exposure. Therefore, we tested the hypothesis that long-term repetitive exposure to HBO would modify the degree of DNA damage. Combat swimmers and underwater demolition team divers were investigated because their diving practice comprises repetitive long-term exposure to HBO over years. Nondiving volunteers with and without endurance training served as controls. In addition to the measurement of DNA damage in peripheral blood (comet assay), blood antioxidant enzyme activities, and the ratio of oxidized and reduced glutathione content, we assessed the DNA damage and superoxide anion radical (O2•−) production induced by a single ex vivo HBO exposure of isolated lymphocytes. All parameters of oxidative stress and antioxidative capacity in vivo were comparable in the four different groups. Exposure to HBO increased both the level of DNA damage and O2•− production in lymphocytes, and this response was significantly more pronounced in the cells obtained from the combat swimmers than in all the other groups. However, in all groups, DNA damage was completely removed within 1 h. We conclude that, at least in healthy volunteers with endurance training, long-term repetitive exposure to HBO does not modify the basal blood antioxidant capacity or the basal level of DNA strand breaks. The increased ex vivo HBO-related DNA damage in isolated lymphocytes from these subjects, however, may reflect enhanced susceptibility to oxidative DNA damage.

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