scholarly journals Insights into Cadmium-Induced Carcinogenesis through an In Vitro Study Using C3H10T1/2Cl8 Cells: The Multifaceted Role of Mitochondria

2021 ◽  
Vol 22 (19) ◽  
pp. 10837
Author(s):  
Monica Oldani ◽  
Anna Maria Villa ◽  
Marta Manzoni ◽  
Pasquale Melchioretto ◽  
Paolo Parenti ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Oxidative Phosphorylation ◽  
Cell Transformation ◽  
In Vitro Study ◽  
Atp Synthesis ◽  
Cadmium Treatment ◽  
Mouse Embryo Fibroblasts

In this paper, we report the metabolic characterization of two foci, F1 and F3, obtained at the end of Cell Transformation Assay (CTA), performed by treating C3H10T1/2Cl8 mouse embryo fibroblasts with 1 μM CdCl2 for 24 h. The elucidation of the cadmium action mechanism can be useful both to improve the in vitro CTA and to yield insights into carcinogenesis. The metabolism of the two foci was investigated through Seahorse and enzyme activity assays; mitochondria were studied in confocal microscopy and reactive oxygen species were detected by flow cytometry. The results showed that F1 focus has higher glycolytic and TCA fluxes compared to F3 focus, and a more negative mitochondrial membrane potential, so that most ATP synthesis is performed through oxidative phosphorylation. Confocal microscopy showed mitochondria crowded in the perinuclear region. On the other hand, F3 focus showed lower metabolic rates, with ATP mainly produced by glycolysis and damaged mitochondria. Overall, our results showed that cadmium treatment induced lasting metabolic alterations in both foci. Triggered by the loss of the Pasteur effect in F1 focus and by mitochondrial impairment in F3 focus, these alterations lead to a loss of coordination among glycolysis, TCA and oxidative phosphorylation, which leads to malignant transformation.

scholarly journals Insights into cadmium-induced carcinogenesis through an in-vitro study using C3H10T1/2Cl8 cells: the multifaceted role of mitochondria

2021 ◽  
Author(s):  
Monica Oldani ◽  
Anna Maria Villa ◽  
Marta Manzoni ◽  
Pasquale Melchioretto ◽  
Paolo Parenti ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Oxidative Phosphorylation ◽  
Cell Transformation ◽  
Expression Patterns ◽  
In Vitro Study ◽  
Atp Synthesis ◽  
Cadmium Treatment

Abstract In this paper we report the metabolic characterization of two foci, F1 and F3, obtained at the end of Cell Transformation Assay (CTA), performed by treating C3H10T1/2Cl8 mouse embryo fibroblasts with 1 µM CdCl2 for 24 h. The elucidation of cadmium action mechanism can be useful both to improve the in vitro CTA and to yield insights into carcinogenesis. We previously showed that, despite being both completely transformed type III foci, F1 and F3 foci display different morphologies, proliferative behaviors and gene expression patterns. In this work, the metabolism of the two foci was investigated through Seahorse and enzyme activity assays; moreover, mitochondria were studied in confocal microscopy and reactive oxygen species were detected by flow cytometry. Results showed that F1 focus has higher glycolytic and TCA fluxes compared to F3 focus, and a more negative mitochondrial membrane potential (Δψ), so that most ATP synthesis is performed through oxidative phosphorylation. Confocal microscopy showed mitochondria crowded in the perinuclear region. On the other hand, F3 focus showed lower metabolic rates, with ATP mainly produced by glycolysis and damaged mitochondria. On the whole, our results showed that cadmium treatment induced lasting metabolic alterations in both foci. Triggered by the loss of Pasteur effect in F1 focus and by mitochondrial impairment in F3 focus, these alterations lead to a loss of coordination among glycolysis, TCA and oxidative phosphorylation, which leads to malignant transformation.

scholarly journals Role of Plant Cell Conditioned Medium in the Phenotypic Expression of Nitrogenase Activity of Rhizobium trifolii Strain T1

1980 ◽  
Vol 33 (5) ◽  
pp. 613 ◽  
Author(s):  
Minocher Reporter ◽  
Mary L Skotnicki ◽  
Barry G Rolfe
Keyword(s):  
Oxidative Phosphorylation ◽  
Conditioned Medium ◽  
Plant Cell ◽  
Nitrogenase Activity ◽  
Phenotypic Expression ◽  
Atp Synthesis ◽  
Atp Production ◽  
Cell Conditioned Medium

The influence of substances from a conditioned medium of cultured plant cells on nitrogenase activity, respiration and ATP synthesis was investigated in R. tri/olii strain Tl. Nitrogenase activity in strain Tl was dependent on the addition of the plant cell conditioned medium. Studies showed that the initial effects of the plant substances on rhizobial cells was to increase their respiration rate and ATP production. Mutants of strain Tl which were uncoupled in their oxidative phosphorylation, were also tested. However, the plant factors had no effect on respiration and ATP synthesis and also failed to elicit in vitro nitrogenase activity in these mutants. It is proposed that these plant factors act by increasing the efficiency of oxidative phosphorylation, making more ATP available, and thus stimulating nitrogenase activity of R. tri/olii cells.

scholarly journals Neutrophils can promote clotting via FXI and impact clot structure via neutrophil extracellular traps in a distinctive manner in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Y. Shi ◽  
J. S. Gauer ◽  
S. R. Baker ◽  
H. Philippou ◽  
S. D. Connell ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Structure Formation ◽  
In Vitro Study ◽  
Neutrophil Extracellular Traps ◽  
Vitro Study ◽  
Extracellular Traps ◽  
Clot Structure

AbstractNeutrophils and neutrophil extracellular traps (NETs) have been shown to be involved in coagulation. However, the interactions between neutrophils or NETs and fibrin(ogen) in clots, and the mechanisms behind these interactions are not yet fully understood. In this in vitro study, the role of neutrophils or NETs on clot structure, formation and dissolution was studied with a combination of confocal microscopy, turbidity and permeation experiments. Factor (F)XII, FXI and FVII-deficient plasmas were used to investigate which factors may be involved in the procoagulant effects. We found both neutrophils and NETs promote clotting in plasma without the addition of other coagulation triggers, but not in purified fibrinogen, indicating that other factors mediate the interaction. The procoagulant effects of neutrophils and NETs were also observed in FXII- and FVII-deficient plasma. In FXI-deficient plasma, only the procoagulant effects of NETs were observed, but not of neutrophils. NETs increased the density of clots, particularly in the vicinity of the NETs, while neutrophils-induced clots were less stable and more porous. In conclusion, NETs accelerate clotting and contribute to the formation of a denser, more lysis resistant clot architecture. Neutrophils, or their released mediators, may induce clotting in a different manner to NETs, mediated by FXI.

scholarly journals In Vitro Effects of Resin Infiltration on Enamel Erosion Inhibition

2015 ◽  
Vol 40 (5) ◽  
pp. 492-502 ◽  
Author(s):  
GC Oliveira ◽  
AP Boteon ◽  
FQ Ionta ◽  
MJ Moretto ◽  
HM Honório ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Artificial Saliva ◽  
In Vitro Study ◽  
Adhesive System ◽  
Dental Erosion ◽  
Deep Penetration ◽  
Resin Infiltration ◽  
Fluorescence Confocal Microscopy ◽  
In Vitro Effects

SUMMARY Resin-based materials that show promising effects for preventing the progression of erosion have been studied. This in vitro study evaluated the effects of applying resin-based materials, including resin infiltration, on previously eroded enamel subjected to erosive challenges. The influence of enamel surface etching prior to application of the material was also studied. Bovine enamel blocks were immersed in hydrochloric acid (HCl), 0.01 M (pH 2.3), for 30 seconds in order to form a softened erosion lesion. The blocks were then randomly divided into nine groups (n=12) and treated as follows: C = control without treatment; Hel = pit & fissure resin sealant (Helioseal Clear); Adh = two-step self-etching adhesive system (AdheSe); Tet = two-step conventional adhesive system (Tetric N-bond); and Inf = infiltrant (Icon). The Helno, Adhno, Tetno, and Infno groups received the same materials without (or with no) surface conditioning. The depth of the material's penetration into softened erosion lesions was qualitatively analyzed using reflection and fluorescence confocal microscopy. After application of the materials, the blocks were immersed in HCl for two minutes; this step was followed by immersion in artificial saliva for 120 minutes four times a day for five days (erosive cycling). Both the enamel alteration and material thickness were analyzed using profilometry, and the results were submitted to Kruskal-Wallis and Dunn tests (p>0.05). Images from the confocal microscopy showed minimal penetration of Adh/Adhno and deep penetration of Inf/Infno into the erosive lesions. The groups Hel, Adh, Inf, Tetno, and Infno resulted in the formation of a layer of material over the enamel, which was effective in inhibiting the progression of erosion. In conclusion, the infiltrant, with or without etching, was able to penetrate and protect the enamel against dental erosion. The other resin-based materials, except for the two-step conventional adhesive, were able to penetrate and inhibit the progression of erosive lesions only when they were applied after enamel etching.

scholarly journals Role of glyoxalases in wound healing process: an in vitro study

2021 ◽  
Vol 165 ◽  
pp. 39
Author(s):  
Francesca Lombardi ◽  
Silvano Santini ◽  
Paola Palumbo ◽  
Valeria Cordone ◽  
Virginio Bignotti ◽  
...  
Keyword(s):  
Wound Healing ◽  
Healing Process ◽  
In Vitro Study ◽  
Vitro Study ◽  
Wound Healing Process

scholarly journals Characterization of a Dye-Decolorizing Peroxidase from Irpex lacteus Expressed in Escherichia coli: An Enzyme with Wide Substrate Specificity Able to Transform Lignosulfonates

2021 ◽  
Vol 7 (5) ◽  
pp. 325
Author(s):  
Laura Isabel de de Eugenio ◽  
Rosa Peces-Pérez ◽  
Dolores Linde ◽  
Alicia Prieto ◽  
Jorge Barriuso ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Veratryl Alcohol ◽  
Dye Decolorization ◽  
Irpex Lacteus ◽  
Type D ◽  
Lignin Peroxidases

A dye-decolorizing peroxidase (DyP) from Irpex lacteus was cloned and heterologously expressed as inclusion bodies in Escherichia coli. The protein was purified in one chromatographic step after its in vitro activation. It was active on ABTS, 2,6-dimethoxyphenol (DMP), and anthraquinoid and azo dyes as reported for other fungal DyPs, but it was also able to oxidize Mn2+ (as manganese peroxidases and versatile peroxidases) and veratryl alcohol (VA) (as lignin peroxidases and versatile peroxidases). This corroborated that I. lacteus DyPs are the only enzymes able to oxidize high redox potential dyes, VA and Mn+2. Phylogenetic analysis grouped this enzyme with other type D-DyPs from basidiomycetes. In addition to its interest for dye decolorization, the results of the transformation of softwood and hardwood lignosulfonates suggest a putative biological role of this enzyme in the degradation of phenolic lignin.

Confocal Microscopy Characterization of In Vitro Tests of Controlled Atmosphere Plasma Spraying Hydroxyapatite (CAPS-HA) Coatings

2003 ◽  
Vol 254-256 ◽  
pp. 319-322 ◽  
Author(s):  
Khiam Aik Khor ◽  
M. Espanol Pons ◽  
Gemma Bertran-Vidal ◽  
Núria Llorca-Isern ◽  
Michel Jeandin ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Plasma Spraying ◽  
In Vitro Tests ◽  
Controlled Atmosphere ◽  
Microscopy Characterization

scholarly journals Genotoxicity and inflammatory potential of stainless steel welding fume particles: an in vitro study on standard vs Cr(VI)-reduced flux-cored wires and the role of released metals

2021 ◽  
Author(s):  
Sarah McCarrick ◽  
Valentin Romanovski ◽  
Zheng Wei ◽  
Elin M. Westin ◽  
Kjell-Arne Persson ◽  
...  
Keyword(s):  
Dna Damage ◽  
In Vitro Study ◽  
Human Monocyte ◽  
Welding Fumes ◽  
Welding Fume ◽  
Increased Risk ◽  
Underlying Mechanisms ◽  
Cored Wires

AbstractWelders are daily exposed to various levels of welding fumes containing several metals. This exposure can lead to an increased risk for different health effects which serves as a driving force to develop new methods that generate less toxic fumes. The aim of this study was to explore the role of released metals for welding particle-induced toxicity and to test the hypothesis that a reduction of Cr(VI) in welding fumes results in less toxicity by comparing the welding fume particles of optimized Cr(VI)-reduced flux-cored wires (FCWs) to standard FCWs. The welding particles were thoroughly characterized, and toxicity (cell viability, DNA damage and inflammation) was assessed following exposure to welding particles as well as their released metal fraction using cultured human bronchial epithelial cells (HBEC-3kt, 5–100 µg/mL) and human monocyte-derived macrophages (THP-1, 10–50 µg/mL). The results showed that all Cr was released as Cr(VI) for welding particles generated using standard FCWs whereas only minor levels (< 3% of total Cr) were released from the newly developed FCWs. Furthermore, the new FCWs were considerably less cytotoxic and did not cause any DNA damage in the doses tested. For the standard FCWs, the Cr(VI) released in cell media seemed to explain a large part of the cytotoxicity and DNA damage. In contrast, all particles caused rather similar inflammatory effects suggesting different underlying mechanisms. Taken together, this study suggests a potential benefit of substituting standard FCWs with Cr(VI)-reduced wires to achieve less toxic welding fumes and thus reduced risks for welders.

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