An Optimized MTT Bioassay for Determination of Cytotoxicity of Fumonisins in Turkey Lymphocytes

1994 ◽  
Vol 77 (2) ◽  
pp. 512-516 ◽  
Author(s):  
Mary A Dombrink-Kurtzman ◽  
Glenn A Bennett ◽  
John L Richard
Keyword(s):  
Gradient Centrifugation ◽  
In Vitro Cytotoxicity ◽  
Tetrazolium Salt ◽  
Animal Testing ◽  
Cytotoxicity Assays ◽  
Cytoplasmic Vacuolization ◽  
Diphenyl Tetrazolium Bromide ◽  
Dose Dependent

Abstract In vitro cytotoxicity assays have been performed for detection and quantitation of fumonisins, as possible alternatives for whole animal testing. This study was undertaken to establish optimal in vitro conditions using turkey lymphocytes. Turkey lymphocytes were isolated from peripheral blood by Percoll gradient centrifugation. Cytotoxicity of fu-monisin B1 (FB1) and B2 (FB2) was determined by exposing lymphocytes to FB1 or FB2 at concentrations of 0.01–25 μg/mL for 24,48, or 72 h at 39°C. The MTT bioassay was used to measure cell viability and proliferation. In metabolically active cells, the tetrazolium salt, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], was reduced to MTT formazan. Turkey lymphocytes that had been exposed in vitro to FB1 and FB2 for 48 and 72 h showed inhibition of cell proliferation that was dose-dependent. The 50% inhibitory dose for FB1 and FB2 was 0.4–5 μg/mL. Cells exposed to FB1 or FB2 exhibited high levels of cytoplasmic vacuolization and were unable to proliferate, whereas proliferation of control lymphocytes was observed at 48 and 72 h. FB2 was 3- to 4-fold more cytotoxic than FB1.

Relationship between quin2-determined cytosolic [Ca2+] and sweat secretion

1988 ◽  
Vol 254 (2) ◽  
pp. C310-C317 ◽  
Author(s):  
K. Sato ◽  
F. Sato
Keyword(s):  
Calcium Concentration ◽  
Gradient Centrifugation ◽  
Co2 Production ◽  
Sweat Glands ◽  
Secretory Cells ◽  
Sweat Secretion ◽  
Dose Dependent

Although both methacholine (MCh)- and A23187-induced sweat secretion are known to be strictly dependent on extracellular Ca2+, the role of intracellular calcium concentration ([Ca2+]i) in eccrine sweating has not been clarified. Partially purified eccrine secretory cells were prepared from 400 to 600 isolated secretory coils of monkey sweat glands by serial collagenase digestion and Percoll gradient centrifugation. The quin2 method was used for semiquantitative determination of [Ca2+]i, MCh increased [Ca2+]i in a dose-dependent, reversible, and pharmacologically specific manner (from the resting [Ca2+]i of 80-320 nM) but failed to increase [Ca2+]i in a Ca2+-free medium. A23187 (10(-7) M) increased [Ca2+]i to approximately 900 nM. Theophylline (TH), isoproterenol (ISO), and forskolin (FK) had no effect on the resting [Ca2+]i but, in combination, attenuated the effect of subsequently added MCh and A23187. A23187 at 10(-7) M failed to stimulate sweat secretion or CO2 production in vitro from the quin2-loaded intact isolated sweat glands and dispersed sweat secretory cells, respectively. The observed dissociation between the increase in [Ca2+] may suggest either that MCh stimulation induces some unknown excitatory signal in addition to a rise in [Ca2+] or that A23187-induced Ca2+ influx into the secretory cells is much lower in undissociated sweat glands.

scholarly journals Synthesis and In Vitro Antitumor Activity of Novel Bivalent β-Carboline-3-carboxylic Acid Derivatives with DNA as a Potential Target

2018 ◽  
Vol 19 (10) ◽  
pp. 3179 ◽  
Author(s):  
Hongling Gu ◽  
Na Li ◽  
Jiangkun Dai ◽  
Yaxi Xi ◽  
Shijun Wang ◽  
...  
Keyword(s):  
Antitumor Activity ◽  
Cell Apoptosis ◽  
Docking Studies ◽  
In Vitro Cytotoxicity ◽  
Dependent Manner ◽  
Cycle Arrest ◽  
Dna Binding Affinity ◽  
Dose Dependent ◽  
Mcf 7

A series of novel bivalent β-carboline derivatives were designed and synthesized, and in vitro cytotoxicity, cell apoptosis, and DNA-binding affinity were evaluated. The cytotoxic results demonstrated that most bivalent β-carboline derivatives exhibited stronger cytotoxicity than the corresponding monomer against the five selected tumor cell lines (A549, SGC-7901, Hela, SMMC-7721, and MCF-7), indicating that the dimerization at the C3 position could enhance the antitumor activity of β-carbolines. Among the derivatives tested, 4B, 6i, 4D, and 6u displayed considerable cytotoxicity against A549 cell line. Furthermore, 4B, 6i, 4D, and 6u induced cell apoptosis in a dose-dependent manner, and caused cell cycle arrest at the S and G2/M phases. Moreover, the levels of cytochrome C in mitochondria, and the expressions of bcl-2 protein, decreased after treatment with β-carbolines, which indicated that 6i and 6u could induce mitochondria-mediated apoptosis. In addition, the results of UV-visible spectral, thermal denaturation, and molecular docking studies revealed that 4B, 6i, 4D, and 6u could bind to DNA mainly by intercalation.

scholarly journals Folate-modified, curcumin and paclitaxel co-loaded PLA-TPGS nanoparticles: preparation, optimization and in vitro cytotoxicity assays

2018 ◽  
Vol 9 (2) ◽  
pp. 025004 ◽  
Author(s):  
Hai Doan Do ◽  
Hao Le Thi ◽  
Thu Huong Le Thi ◽  
Hoai Nam Nguyen ◽  
Van Khanh Bui ◽  
...  
Keyword(s):  
In Vitro Cytotoxicity ◽  
Cytotoxicity Assays

High-resolution capillary electrophoresis for the determination of carbamylated albumin

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Julien Favresse ◽  
Joris Delanghe
Keyword(s):  
Separation Efficiency ◽  
Theoretical Plate ◽  
Clinical Routine ◽  
Negative Prognostic Factor ◽  
Symmetry Factor ◽  
Primary Amino ◽  
The Mean ◽  
Dose Dependent

Abstract Objectives Carbamylation is a non-enzymatic post-translational reaction of a primary amino group of a protein with isocyanate. The albumin carbamylation is a negative prognostic factor in chronic kidney disease (CKD) patients and induce charge difference implying an observed shift in electrophoretic mobility that can be measured through a symmetry factor (SF). Methods The Helena V8 and the Sebia Capillarys 2 systems were used for all experiments. The effect of in vitro carbamylation on the SF by spiking increasing concentrations of potassium isocyanate (KCNO) in serum of three healthy volunteers was investigated. Theoretical plate numbers (N) as a surrogate of separation efficiency were also calculated and correlations between SF and renal function biomarkers were performed on 284 patients. Results A dose-dependent impact of KCNO on the SF was observed for both methods with the Helena V8 being more sensitive. The mean N was significantly higher on the Helena V8 as compared to the Sebia Capillarys 2 (2,972 vs. 444.1, p<0.0001). The SF correlated significantly with eGFR (r=0.50, p<0.0001), creatinine (r=−0.31, p<0.0001) and urea (r=−0.34, p<0.0001) on the Helena V8. On the Sebia Capillarys 2, a significant correlation was only observed with eGFR (r=0.17, p=0.004). A better discrimination between CKD stages was also observed using the Helena V8. Conclusions Thanks to a higher mean N, the Helena V8 might offer new possibilities, including detection of carbamylated albumin through SF calculation. Further studies are still needed to confirm the interest of using this type of assays in clinical routine.

scholarly journals Punicalagin Regulates Key Processes Associated with Atherosclerosis in THP-1 Cellular Model

2020 ◽  
Vol 13 (11) ◽  
pp. 372
Author(s):  
Sanaa Almowallad ◽  
Etimad Huwait ◽  
Rehab Al-Massabi ◽  
Salma Saddeek ◽  
Kalamegam Gauthaman ◽  
...  
Keyword(s):  
Cholesterol Efflux ◽  
Cell Model ◽  
In Vitro Cytotoxicity ◽  
Intercellular Adhesion Molecule ◽  
Potential Candidate ◽  
Monocyte Chemoattractant Protein 1 ◽  
Cytotoxicity Assays ◽  
Monocyte Migration ◽  
Ifn Γ

Atherosclerosis may lead to cardiovascular diseases (CVD), which are the primary cause of death globally. In addition to conventional therapeutics for CVD, use of nutraceuticals that prevents cholesterol deposition, reduce existing plaques and hence anti-atherosclerotic effects of nutraceuticals appeared to be promising. As such, in the present study we evaluated the beneficial effects of punicalagin, a phytochemical against an atherosclerotic cell model in vitro. Cytotoxicity assays were examined for 10 µM concentration of punicalagin on THP-1 macrophages. Real-time-polymerase chain reaction (RT-PCR) was used to analyze monocyte chemoattractant protein-1 (MCP-1) and Intercellular adhesion molecule (ICAM-1) expressions. Monocyte migration and cholesterol efflux assays were performed to investigate punicalagin’s further impact on the key steps of atherosclerosis. Cytotoxicity assays demonstrated no significant toxicity for punicalagin (10 µM) on THP-1 macrophages. Punicalagin inhibited the IFN-γ-induced overexpression of MCP-1 and ICAM-1 in macrophages by 10 fold and 3.49 fold, respectively, compared to the control. Punicalagin also reduced the MCP-1- mediated migration of monocytes by 28% compared to the control. Percentages of cellular cholesterol efflux were enhanced in presence or absence of IFN-γ by 88% and 84% compared to control with 58% and 62%, respectively. Punicalagin possesses anti-inflammatory and anti-atherosclerotic effects. Punicalagin also did not exhibit any cytotoxicity and therefore can be considered a safe and potential candidate for the treatment and prevention of atherosclerosis.

A Comparison of In Vitro Cytotoxicity Assays and Their Application to Water Samples

1987 ◽  
Vol 15 (1) ◽  
pp. 20-29 ◽  
Author(s):  
Stephen M. Hunt ◽  
Christina Chrzanowska ◽  
Christopher R. Barnett ◽  
Helen N. Brand ◽  
John K. Fawell
Keyword(s):  
Cell Growth ◽  
Water Samples ◽  
Industrial Effluent ◽  
Exposure Period ◽  
In Vitro Cytotoxicity ◽  
Cloning Efficiency ◽  
Cytotoxicity Assays ◽  
River Water Samples ◽  
Vital Dye

A group of 13 compounds were tested for in vitro cytotoxicity in four test systems; MIT-24 test, inhibition of cell growth (protein method), inhibition of cell growth (vital dye method) and cloning efficiency. In general, all four assays tended to rank compounds in a similar order for toxicity. The length of the exposure period appeared to be important for some compounds. The cytotoxicity of a variety of water samples was examined in two tests; inhibition of cell growth (vital dye method) and cloning efficiency. Under the conditions in which the assays were carried out, the latter proved to be the more sensitive test. River water samples gave little or no indication of cytotoxicity, samples of domestic sewage effluent gave some evidence of cytotoxicity, while an industrial effluent was markedly cytotoxic.

scholarly journals Synthesis and Properties of CurNQ for the Theranostic Application in Ovarian Cancer Intervention

Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4471
Author(s):  
Lara G. Freidus ◽  
Pradeep Kumar ◽  
Thashree Marimuthu ◽  
Priyamvada Pradeep ◽  
Viness Pillay ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cancer Detection ◽  
Ovarian Cancer Cell ◽  
In Vitro Cytotoxicity ◽  
Cancer Targeting ◽  
The Novel ◽  
Cytotoxicity Assays ◽  
Ovarian Cancer Cell Lines ◽  
Cancer Intervention

Synthesis of a novel theranostic molecule for targeted cancer intervention. A reaction between curcumin and lawsone was carried out to yield the novel curcumin naphthoquinone (CurNQ) molecule (2,2′-((((1E,3Z,6E)-3-hydroxy-5-oxohepta-1,3,6-triene-1,7-diyl) bis(2-methoxy-4,1-phenylene))bis(oxy))bis(naphthalene-1,4-dione). CurNQ’s structure was elucidated and was fully characterized. CurNQ was demonstrated to have pH specific solubility, its saturation solubility increased from 11.15 µM at pH 7.4 to 20.7 µM at pH 6.8. This pH responsivity allows for cancer targeting (Warburg effect). Moreover, CurNQ displayed intrinsic fluorescence, thus enabling imaging and detection applications. In vitro cytotoxicity assays demonstrated the chemotherapeutic properties of CurNQ as CurNQ reduced cell viability to below 50% in OVCAR-5 and SKOV3 ovarian cancer cell lines. CurNQ is a novel theranostic molecule for potential targeted cancer detection and treatment.

Determination of mosquito bloodmeal pH in situ by ion-selective microelectrode measurement: implications for the regulation of malarial gametogenesis

Parasitology ◽  
2000 ◽  
Vol 120 (6) ◽  
pp. 547-551 ◽  
Author(s):  
O. BILLKER ◽  
A. J. MILLER ◽  
R. E. SINDEN
Keyword(s):  
Xanthurenic Acid ◽  
Mosquito Vector ◽  
Dependent Manner ◽  
Ph Increase ◽  
Ph Range ◽  
Dose Dependent

Malarial gametocytes circulate in the peripheral blood of the vertebrate host as developmentally arrested intra-erythrocytic cells, which only resume development into gametes when ingested into the bloodmeal of the female mosquito vector. The ensuing development encompasses sexual reproduction and mediates parasite transmission to the insect. In vitro the induction of gametogenesis requires a drop in temperature and either a pH increase from physiological blood pH (ca pH 7·4) to about pH 8·0, or the presence of a gametocyte-activating factor recently identified as xanthurenic acid (XA). However, it is unclear whether either the pH increase or XA act as natural triggers in the mosquito bloodmeal. We here use pH-sensitive microelectrodes to determine bloodmeal pH in intact mosquitoes. Measurements taken in the first 30 min after ingestion, when malarial gametogenesis is induced in vivo, revealed small pH increases from 7·40 (mouse blood) to 7·52 in Aedes aegypti and to 7·58 in Anophěles stephensi. However, bloodmeal pH was clearly suboptimal if compared to values required to induce gametogenesis in vitro. Xanthurenic acid is shown to extend the pH-range of exflagellation in vitro in a dose-dependent manner to values that we have observed in the bloodmeal, suggesting that in vivo malarial gametogenesis could be further regulated by both these factors.

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