Search results for " CELL CULTURES"

GSH: A MARKER FOR OXIDATIVE STRESS IN HUMAN CELL CULTURES

2013

GSH: A MARKER FOR OXIDATIVE STRESS IN HUMAN CELL CULTURES Gueli Maria Concetta Dipartimento di Biomedicina Sperimentale e Neuroscienze Cliniche (BioNEC), Università degli Studi di Palermo. Reactive oxygen species (ROS) play an important role in physiological processes, but when being in excess ROS react readily with proteins, lipids, carbohydrates, and nucleic acids, often inducing irreversible functional alterations or even complete destruction. In cells, under physiological conditions, the production and detoxification of ROS are more or less balanced. GSH functions as antioxidant and the oxidative conversion of GSH to GSSG is widely recognized as a reliable index of oxidative stress. The…

Quality controls for cell cultures: identification of interspecies cross-contamination by PCR-RFLP analysis of the cytochrome b gene

2012

Cross-contaminations of a cell line with cells of different species represent a potential risk in laboratories handling human and animal cells. Therefore, it is necessary to control such contaminations. Tests based on mitochondrial DNA (mtDNA) are used in forensic analysis, phylogenetic studies and in food authentication. However, the use of mtDNA in quality controls of cell cultures is recent. Mitochondrial sequence differences of closely related animal species are five- to tenfold higher than those of nuclear genes. On the contrary, intraspecies variation in mitochondrial sequences is low in most animal species. Moreover, each cell contains 100–10.000 mitochondrial genomes. The amount of …

EFFECTS OF CELLULAR SHORT-TERM STARVATION ON CONVENTIONAL CHEMOTHERAPY RESPONSE IN HUMAN CANCER: UNDERSTANDING OF MOLECULAR MECHANISMS AND MICRORNAS …

Background: Short Term Starvation (STS) is a type of dietary restriction able to reduce tumorigenesis and cancer progression but molecular bases of this effect are still unclear. Aim: In vitro analysis of STS effects in presence of chemotherapy and evaluation of microRNAs (miRNAs) involvement. Results: STS affects the expression profiles of miRNAs involved in chemotherapy response leading to cancer cells sensitization and to healthy cells protection.

The use of three-dimensional oral mucosa cell cultures to assess the toxicity of soldered and welded wires

2007

The aim of the present study was to determine whether there is a difference in toxicity and loss of viability of three-dimensional (3D) reconstructed human oral epithelium (RHOE) cell cultures induced by point-welded (PW), laser-welded (LW), and silver-soldered (SiS) orthodontic wires. Three types of soldered stainless steel (SS) wires: PW, LW, and SiS were prepared ( n = 3) and subjected to multiple end-point analysis (MEA). Six pieces were cut from each wire. Each piece was placed on the triplicate cell cultures (RHOE model based on TR 146 cells). After 24 hours of topical exposure, the cell cultures were cut and stained with haematoxylin/eosin. Toxicity was assessed by evaluating the mor…

Prenatal exposure to the CB1 receptor agonist WIN 55,212-2 causes learning disruption associated with impaired cortical NMDA receptor function and em…

2005

The aim of this study was to investigate whether prenatal exposure to the cannabinoid CB1 receptor agonist WIN 55,212-2 (WIN) at a daily dose devoid of overt signs of toxicity and/or gross malformations (0.5 mg/kg, gestation days 5-20), influences cortical glutamatergic neurotransmission, learning and emotional reactivity in rat offspring. Basal and K+-evoked extracellular glutamate levels were significantly lower in cortical cell cultures obtained from pups exposed to WIN during gestation with respect to those measured in cultures obtained from neonates born from vehicle-treated dams. The addition of NMDA to cortical cell cultures from neonates born from vehicle-treated dams concentration-…

Toxicity of used orthodontic archwires assessed by three-dimensional cell culture

2006

The aim of the present study was to determine whether used orthodontic wires made of different materials cause toxicity and loss of viability on three-dimensional (3D) cell cultures. Three types of orthodontic wires, stainless steel, Nitinol, and TMA (n = 9) which had been used clinically in fixed appliances for a period of 1 month, were retrieved at random from five patients. Both upper and lower archwires were collected and subjected to two different protocols: to assess toxicity, two pieces of each wire were placed on 3D cell cultures (reconstituted human epithelium); to investigate the possibility of cell damage, the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) ass…

A Ciliary Motility Index for Activity Measurement in Cell Cultures With Respiratory Syncytial Virus

2018

[EN] Background: The respiratory epithelium is frequently infected by the respiratory syncytial virus, resulting in inflammation, a reduction in cilia activity and an increase in the production of mucus. Methods: In this study, an automatic method has been proposed to characterize the ciliary motility from cell cultures by means of a motility index using a dense optical flow algorithm. This method allows us to determine the ciliary beat frequency (CBF) together with a ciliary motility index of the cells in the cultures. The object of this analysis is to automatically distinguish between normal and infected cells in a culture. Results: The method was applied in 2 stages. It was concluded fro…

Expression and developmental regulation of the cystine/glutamate exchanger (xc-) in the rat.

2007

The cystine/glutamate exchanger (antiporter x c − ) is a membrane transporter involved in the uptake of cystine, the rate-limiting amino acid in the synthesis of glutathione. Recent studies suggest that the antiporter plays a role in the slow oxidative excitotoxity and in the pathological effects of β-N-oxalylamino-l-alanine, the molecule responsible for neurolathyrism, a neurotoxic upper motor neuron disease. The mouse cystine/glutamate exchanger has been cloned and showed to be composed of two distinct proteins, one of which being a novel protein, named xCT, of 502 amino acids and 12 putative trans-membrane domains. We have generated and purified a polyclonal antibody to mouse xCT and stu…

Neuronal and BBB damage induced by sera from patients with secondary progressive multiple sclerosis.

2009

An important component of the pathogenic process of multiple sclerosis (MS) is the blood-brain barrier (BBB) damage. We recently set an in vitro model of BBB, based on a three-cell-type co-culture system, in which rat neurons and astrocytes synergistically induce brain capillary endothelial cells to form a monolayer with permeability properties resembling those of the physiological BBB. Herein we report that the serum from patients with secondary progressive multiple sclerosis (SPMS) has a damaging effect on isolated neurons. This finding suggests that neuronal damaging in MS could be a primary event and not only secondary to myelin damage, as generally assumed. SPMS serum affects the perme…

Isomer-nonspecific action of dichlorodiphenyltrichloroethane on aryl hydrocarbon receptor and G-protein-coupled receptor 30 intracellular signaling i…

2014

Abstract Extended residual persistence of the pesticide dichlorodiphenyltrichloroethane (DDT) raises concerns about its long-term neurotoxic effects. Little is known, however, about DDT toxicity during the early stages of neural development. This study demonstrated that DDT-induced apoptosis of mouse embryonic neuronal cells is a caspase-9-, caspase-3-, and GSK-3β-dependent process, which involves p,p’ -DDT-specific impairment of classical ERs. It also provided evidence for DDT-isomer-nonspecific alterations of AhR- and GPR30-mediated intracellular signaling, including changes in the levels of the receptor and receptor-regulated mRNAs, and also changes in the protein levels of the receptors…