Are polyphenols antioxidants or pro-oxidants? What do we learn from cell culture and in vivo studies?

2008 ◽  
Vol 476 (2) ◽  
pp. 107-112 ◽  
Author(s):  
Barry Halliwell
Keyword(s):  
Cell Culture ◽  
In Vivo Studies

scholarly journals Which cytochrome P450 metabolizes phenazepam? Step by step in silico, in vitro, and in vivo studies

2018 ◽  
Vol 33 (2) ◽  
pp. 65-73 ◽  
Author(s):  
Dmitriy V. Ivashchenko ◽  
Anastasia V. Rudik ◽  
Andrey A. Poloznikov ◽  
Sergey V. Nikulin ◽  
Valeriy V. Smirnov ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cytochrome P450 ◽  
In Silico ◽  
In Vivo Studies ◽  
Cyp3a Activity ◽  
Cell Culture Study ◽  
Study Results ◽  
Three Stages

Abstract Background: Phenazepam (bromdihydrochlorphenylbenzodiazepine) is the original Russian benzodiazepine tranquilizer belonging to 1,4-benzodiazepines. There is still limited knowledge about phenazepam’s metabolic liver pathways and other pharmacokinetic features. Methods: To determine phenazepam’s metabolic pathways, the study was divided into three stages: in silico modeling, in vitro experiment (cell culture study), and in vivo confirmation. In silico modeling was performed on the specialized software PASS and GUSAR to evaluate phenazepam molecule affinity to different cytochromes. The in vitro study was performed using a hepatocytes’ cell culture, cultivated in a microbioreactor to produce cytochrome P450 isoenzymes. The culture medium contained specific cytochrome P450 isoforms inhibitors and substrates (for CYP2C9, CYP3A4, CYP2C19, and CYP2B6) to determine the cytochrome that was responsible for phenazepam’s metabolism. We also measured CYP3A activity using the 6-betahydroxycortisol/cortisol ratio in patients. Results: According to in silico and in vitro analysis results, the most probable metabolizer of phenazepam is CYP3A4. By the in vivo study results, CYP3A activity decreased sufficiently (from 3.8 [95% CI: 2.94–4.65] to 2.79 [95% CI: 2.02–3.55], p=0.017) between the start and finish of treatment in patients who were prescribed just phenazepam. Conclusions: Experimental in silico and in vivo studies confirmed that the original Russian benzodiazepine phenazepam was the substrate of CYP3A4 isoenzyme.

scholarly journals Tumor Gene Targeting Using Microspheres: Cell Culture and in vivo Studies

Drug Delivery ◽  
1997 ◽  
Vol 4 (4) ◽  
pp. 263-267 ◽  
Author(s):  
Crispin R. Dass ◽  
E. E. Decruz ◽  
T. L. Walker ◽  
M. A. Burton
Keyword(s):  
Cell Culture ◽  
Gene Targeting ◽  
In Vivo Studies ◽  
Tumor Gene

scholarly journals The Drosophila tango gene encodes a bHLH-PAS protein that is orthologous to mammalian Arnt and controls CNS midline and tracheal development

Development ◽  
1997 ◽  
Vol 124 (22) ◽  
pp. 4571-4582 ◽  
Author(s):  
M. Sonnenfeld ◽  
M. Ward ◽  
G. Nystrom ◽  
J. Mosher ◽  
S. Stahl ◽  
...  
Keyword(s):  
Cell Culture ◽  
Gene Dosage ◽  
Cell Lineage ◽  
Regulatory System ◽  
In Vivo Studies ◽  
Enhancer Element ◽  
Evolutionarily Conserved ◽  
Cns Midline ◽  
Functionally Diverse

The Drosophila single-minded and trachealess bHLH-PAS genes control transcription and development of the CNS midline cell lineage and tracheal tubules, respectively. We show that Single-minded and Trachealess activate transcription by forming dimers with the Drosophila Tango protein that is an orthologue of the mammalian Arnt protein. Both cell culture and in vivo studies show that a DNA enhancer element acts as a binding site for both Single-minded::Tango and Trachealess::Tango heterodimers and functions in controlling CNS midline and tracheal transcription. Isolation and analysis of tango mutants reveal CNS midline and tracheal defects, and gene dosage studies demonstrate in vivo interactions between single-minded::tango and trachealess::tango. These experiments support the existence of an evolutionarily conserved, functionally diverse bHLH-PAS protein regulatory system.

Tissue engineering: a new frontier in physiological genomics

2007 ◽  
Vol 32 (1) ◽  
pp. 28-32 ◽  
Author(s):  
Matthew C. Petersen ◽  
Jozef Lazar ◽  
Howard J. Jacob ◽  
Tetsuro Wakatsuki
Keyword(s):  
Cell Culture ◽  
High Throughput Screening ◽  
3D Culture ◽  
3D Cell Culture ◽  
Cardiac Cells ◽  
In Vivo Studies ◽  
Specific Cell ◽  
Organ Systems ◽  
Engineered Tissue

Considerable progress has been made in the last decade in the engineering and construction of a number of artificial tissue types. These constructs are typically viewed from the perspective of possible sources for implant and transplant materials in the clinical arena. However, incorporation of engineered tissues, often referred to as three-dimensional (3D) cell culture, also offers the possibility for significant advancements in research for physiological genomics. These 3D systems more readily mimic the in vivo setting than traditional 2D cell culture, and offer distinct advantages over the in vivo setting for some organ systems. As an example, cardiac cells in 3D culture 1) are more accessible for siRNA studies, 2) can be engineered with specific cell types, and 3) offer the potential for high-throughput screening of gene function. Here the state-of-the-art is reviewed and the applications for engineered tissue in genomics research are proposed. The ability to use engineered tissue in combination with genomics creates a bridge between traditional cellular and in vivo studies that is critical to enabling the transition of genetic information into mechanistic understanding of disease processes.

scholarly journals Biomaterial-Assisted Regenerative Medicine

2021 ◽  
Vol 22 (16) ◽  
pp. 8657
Author(s):  
Teruki Nii ◽  
Yoshiki Katayama
Keyword(s):  
Cell Culture ◽  
Regenerative Medicine ◽  
Drug Screening ◽  
Drug Research ◽  
Drug Evaluation ◽  
Cell Activity ◽  
The Body ◽  
In Vivo Studies

This review aims to show case recent regenerative medicine based on biomaterial technologies. Regenerative medicine has arousing substantial interest throughout the world, with “The enhancement of cell activity” one of the essential concepts for the development of regenerative medicine. For example, drug research on drug screening is an important field of regenerative medicine, with the purpose of efficient evaluation of drug effects. It is crucial to enhance cell activity in the body for drug research because the difference in cell condition between in vitro and in vivo leads to a gap in drug evaluation. Biomaterial technology is essential for the further development of regenerative medicine because biomaterials effectively support cell culture or cell transplantation with high cell viability or activity. For example, biomaterial-based cell culture and drug screening could obtain information similar to preclinical or clinical studies. In the case of in vivo studies, biomaterials can assist cell activity, such as natural healing potential, leading to efficient tissue repair of damaged tissue. Therefore, regenerative medicine combined with biomaterials has been noted. For the research of biomaterial-based regenerative medicine, the research objective of regenerative medicine should link to the properties of the biomaterial used in the study. This review introduces regenerative medicine with biomaterial.

Impairment of autophagy by TTR V30M aggregates: in vivo reversal by TUDCA and curcumin

2016 ◽  
Vol 130 (18) ◽  
pp. 1665-1675 ◽  
Author(s):  
Cristina A. Teixeira ◽  
Maria do Rosário Almeida ◽  
Maria João Saraiva
Keyword(s):  
Cell Culture ◽  
Amyloid Fibrils ◽  
Transgenic Animals ◽  
Cellular Mechanism ◽  
In Vivo Studies ◽  
Autophagic Flux ◽  
Gi Tract ◽  
Alternative Therapeutic Approach

Transthyretin (TTR)-related amyloidoses are diseases characterized by extracellular deposition of amyloid fibrils and aggregates in tissues composed of insoluble misfolded TTR that becomes toxic. Previous studies have demonstrated the ability of small compounds in preventing and reversing TTR V30M deposition in transgenic mice gastrointestinal (GI) tract as well as lowering biomarkers associated with cellular stress and apoptotic mechanisms. In the present study we aimed to study TTR V30M aggregates effect in autophagy, a cellular mechanism crucial for cell survival that has been implicated in the development of several neurodegenerative diseases. We were able to demonstrate in cell culture that TTR V30M aggregates cause a partial impairment of the autophagic machinery as shown by p62 accumulation, whereas early steps of the autophagic flux remain unaffected as shown by autophagosome number evaluation and LC3 turnover assay. Our studies performed in TTR V30M transgenic animals demonstrated that tauroursodeoxycholic acid (TUDCA) and curcumin effectively reverse p62 accumulation in the GI tract pointing to the ability of both compounds to modulate autophagy additionally to mitigate apoptosis. Overall, our in vitro and in vivo studies establish an association between TTR V30M aggregates and autophagy impairment and suggest the use of autophagy modulators as an additional and alternative therapeutic approach for the treatment of TTR V30M-related amyloidosis.

Resistance to Fas-induced apoptosis in hepatocytes: role of GSH depletion by cell isolation and culture

2002 ◽  
Vol 283 (3) ◽  
pp. G709-G718 ◽  
Author(s):  
Lina Musallam ◽  
Chantal Éthier ◽  
Pierre Selim Haddad ◽  
Francine Denizeau ◽  
Marc Bilodeau
Keyword(s):  
Cell Culture ◽  
Cell Isolation ◽  
In Vivo Studies ◽  
Intracellular Glutathione ◽  
Mouse Hepatocytes ◽  
Induced Apoptosis ◽  
The Relationship ◽  
Apoptotic Induction

The involvement of reduction/oxidation (redox) state in cell sensitivity to apoptosis has been suggested by several studies in which induction of apoptosis was shown to require oxidative stress or GSH extrusion. On the other hand, biochemical studies of caspases revealed that their activation necessitates a reduced cysteine in their active site. This is ensured by maintaining intact intracellular glutathione status during apoptotic induction as reported by in vivo studies. Therefore, we investigated the relationship between intracellular glutathione levels and the sensitivity of mouse hepatocytes in culture to Fas-induced apoptosis as well as potential mechanisms responsible for this sensitivity. We found that total and reduced glutathione levels are decreased by one-half after cell isolation procedure and further decline by 25% during cell culture for 2 h in normal Williams' E medium. Cell culture in medium supplemented with cysteine and methionine maintains glutathione at a level similar to that measured just after cell isolation. Results show that the capacity of Fas to activate caspase-8 and to induce apoptosis requires important intracellular glutathione levels and high GSH/total glutathione ratio. In conclusion, the present study shows that intracellular glutathione plays an important role in maintaining the apoptotic machinery functional and is thus capable of transmitting the apoptotic signal.

scholarly journals Characterization of the Prion Protein Binding Properties of Antisense Oligonucleotides

Biomolecules ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 1 ◽  
Author(s):  
Andrew G. Reidenbach ◽  
Eric Vallabh Minikel ◽  
Hien T. Zhao ◽  
Stacy G. Guzman ◽  
Alison J. Leed ◽  
...  
Keyword(s):  
Cell Culture ◽  
Prion Protein ◽  
Isothermal Titration Calorimetry ◽  
Prion Disease ◽  
Antisense Oligonucleotides ◽  
In Vivo Studies ◽  
Titration Calorimetry ◽  
Independent Manner

Antisense oligonucleotides (ASOs) designed to lower prion protein (PrP) expression in the brain through RNase H1-mediated degradation of PrP RNA are in development as prion disease therapeutics. ASOs were previously reported to sequence-independently interact with PrP and inhibit prion accumulation in cell culture, yet in vivo studies using a new generation of ASOs found that only PrP-lowering sequences were effective at extending survival. Cerebrospinal fluid (CSF) PrP has been proposed as a pharmacodynamic biomarker for trials of such ASOs, but is only interpretable if PrP lowering is indeed the relevant mechanism of action in vivo and if measurement of PrP is unconfounded by any PrP–ASO interaction. Here, we examine the PrP-binding and antiprion properties of ASOs in vitro and in cell culture. Binding parameters determined by isothermal titration calorimetry were similar across all ASOs tested, indicating that ASOs of various chemistries bind full-length recombinant PrP with low- to mid-nanomolar affinity in a sequence-independent manner. Nuclear magnetic resonance, dynamic light scattering, and visual inspection of ASO–PrP mixtures suggested, however, that this interaction is characterized by the formation of large aggregates, a conclusion further supported by the salt dependence of the affinity measured by isothermal titration calorimetry. Sequence-independent inhibition of prion accumulation in cell culture was observed. The inefficacy of non-PrP-lowering ASOs against prion disease in vivo may be because their apparent activity in vitro is an artifact of aggregation, or because the concentration of ASOs in relevant compartments within the central nervous system (CNS) quickly drops below the effective concentration for sequence-independent antiprion activity after bolus dosing into CSF. Measurements of PrP concentration in human CSF were not impacted by the addition of ASO. These findings support the further development of PrP-lowering ASOs and of CSF PrP as a pharmacodynamic biomarker.

scholarly journals Cytotoxicity Studies on Naproxen and Piroxicam Nanoformulations

2020 ◽  
Vol 8 (3) ◽  
pp. 87-94
Author(s):  
Sandeep Patnaik ◽  
K Madhusudhana Rao ◽  
Vijay Sai
Keyword(s):  
Cell Culture ◽  
Cell Viability ◽  
Cultured Cells ◽  
In Vitro Models ◽  
In Vivo Studies ◽  
Tetrazolium Salt ◽  
Mucous Layer ◽  
Cytotoxicity Studies

Caco-2 cells were used as in vitro models to assess the cell viability characteristics of the carriers Soluplus®, Gelucire 50/13 and PVP K25 and the nanoformulations of Naproxen and Piroxicam. The assessment of cell viability was done using the tetrazolium salt based MTT assay. Gelucire 50/13 and its NFs were observed to have slightly higher cytotoxicity than PVP and Soluplus® and their respective NFs. All the NFs were observed to follow the cytotoxicity trend of the polymers. Our results show that no significant decrease in cell viability was seen until 0.01% concentration of Gelucire 50/13 for 12-h exposure. The NFs as well as the polymers alone had no significant effect on the viability of Caco-2 cells below 0.01% concentrations. The intestine has a protective mucous layer, whereas the cell culture monolayers do not. The intestinal tissues also have more capacity to recover from trauma than the cultured cells. Hence the present NFs can be expected to show lesser cytotoxicity when subjected to in vivo studies.  

A T.E.M. study of mouse mammary epithelial cell secretory differentiation cultured on collagen and Matrigel

1991 ◽  
Vol 49 ◽  
pp. 188-189
Author(s):  
W.N. Bentham ◽  
V. Rocha
Keyword(s):  
Cell Culture ◽  
Mammary Epithelial ◽  
Secretory Process ◽  
Cell Culture System ◽  
Protein Maturation ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Mouse Mammary Epithelial Cell ◽  
Secretory Differentiation

It has been an interest of our lab to develop a mammary epethelial cell culture system that faithfully duplicates the in vivo condition of the lactating gland. Since the introduction of collagen as a matrix on which cells are cultivated other E.C.M. type matrices have been made available and are used in many cell culture techniques. We have previously demonstrated that cells cultured on collagen and Matrigel do not differentiate as they do in vivo. It seems that these cultures often produce cells that show a disruption in the secretory process. The appearance of large ribosomal studded vesicles, that specifically label with antibody to casein, suggest an interruption of both protein maturation and secretion at the E.R. to golgi transition. In this report we have examined cultures on collagen and Matrigel at relative high and low seeding densities and compared them to cells from the in vivo condition.

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