scholarly journals Deferoxamine but Not Dimethyloxalylglycine, L-Mimosine, or Cobalt Dichloride Can Interfere with the MTT Assay

2018 ◽  
Vol 2018 ◽  
pp. 1-6
Author(s):  
Anna Sonja Müller ◽  
Klara Janjić ◽  
Gunpreet Oberoi ◽  
Manuela Pensch ◽  
Christoph Kurzmann ◽  
...  
Keyword(s):  
Mtt Assay ◽  
Positive Influence ◽  
Controlled Delivery ◽  
Cellular Functions ◽  
Control Groups ◽  
Experimental Approaches ◽  
Fetal Bovine ◽  
Cobalt Dichloride ◽  
High Concentrations ◽  
Phosphate Buffered Saline

Hypoxia mimetic agents (HMAs) have been shown to have a positive influence on cellular functions in a multitude of tissue regenerative strategies. Novel experimental approaches use biomaterials as carriers for controlled delivery of these HMAs. Here, the cytotoxic aspects of biocompatibility are of key relevance. The MTT assay is widely used to evaluate cytotoxicity and proliferation. Based on the implications from the proceeding research we hypothesized that specific HMAs such as deferoxamine at high concentrations can interfere with the MTT assay. Thus, the aim of this study was to test the repercussions of the HMAs dimethyloxalylglycine, deferoxamine, L-mimosine, and CoCl2 on the validity of the MTT assay. Murine MC3T3-E1 cells were cultured in serum-free alphaMEM and in alphaMEM supplemented with 10 % fetal bovine serum with the HMAs dimethyloxalylglycine, deferoxamine, L-mimosine, and CoCl2, respectively, at 3 mM-0.3 mM for 24 h (experimental groups). Cells without HMAs served as control (control groups). The same experiments were performed with medium and phosphate buffered saline (PBS) without cells. In all settings MTT solution was added to PBS-washed or unwashed culture plates for the last two hours of the incubation period. Then MTT solution was removed and dimethyl sulfoxide was added to dissolve the formazan crystals and absorption was measured. Our data show that the presence of deferoxamine can interfere with the MTT assay if not removed before the addition of MTT. This is particularly important when evaluating cell viability in setups where deferoxamine-loaded biomaterials are used.

scholarly journals In Vivo Toxicity Assessment of Chitosan-Coated Lignin Nanoparticles in Embryonic Zebrafish (Danio rerio)

Nanomaterials ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 111
Author(s):  
Jared S. Stine ◽  
Bryan J. Harper ◽  
Cathryn G. Conner ◽  
Orlin D. Velev ◽  
Stacey L. Harper
Keyword(s):  
Antimicrobial Properties ◽  
Toxicity Assessment ◽  
Environmental Footprint ◽  
Control Groups ◽  
Charged Surfaces ◽  
Chorionic Membrane ◽  
High Concentrations ◽  
And Control ◽  
Oppositely Charged

Lignin is the second most abundant biopolymer on Earth after cellulose. Since lignin breaks down in the environment naturally, lignin nanoparticles may serve as biodegradable carriers of biocidal actives with minimal environmental footprint compared to conventional antimicrobial formulations. Here, a lignin nanoparticle (LNP) coated with chitosan was engineered. Previous studies show both lignin and chitosan to exhibit antimicrobial properties. Another study showed that adding a chitosan coating can improve the adsorption of LNPs to biological samples by electrostatic adherence to oppositely charged surfaces. Our objective was to determine if these engineered particles would elicit toxicological responses, utilizing embryonic zebrafish toxicity assays. Zebrafish were exposed to nanoparticles with an intact chorionic membrane and with the chorion enzymatically removed to allow for direct contact of particles with the developing embryo. Both mortality and sublethal endpoints were analyzed. Mortality rates were significantly greater for chitosan-coated LNPs (Ch-LNPs) compared to plain LNPs and control groups. Significant sublethal endpoints were observed in groups exposed to Ch-LNPs with chorionic membranes intact. Our study indicated that engineered Ch-LNP formulations at high concentrations were more toxic than plain LNPs. Further study is warranted to fully understand the mechanisms of Ch-LNP toxicity.

The Candida albicans plasma membrane protein Rch1p, a member of the vertebrate SLC10 carrier family, is a novel regulator of cytosolic Ca2+ homoeostasis

2012 ◽  
Vol 444 (3) ◽  
pp. 497-502 ◽  
Author(s):  
Linghuo Jiang ◽  
Joerg Alber ◽  
Jihong Wang ◽  
Wei Du ◽  
Xuexue Yang ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Candida Albicans ◽  
Plasma Membrane ◽  
Membrane Protein ◽  
Solute Carrier Family ◽  
Plasma Membrane Protein ◽  
Cellular Functions ◽  
Downstream Target ◽  
Solute Carrier ◽  
High Concentrations

Candida albicans RCH1 (regulator of Ca2+ homoeostasis 1) encodes a protein of ten TM (transmembrane) domains, homologous with human SLC10A7 (solute carrier family 10 member 7), and Rch1p localizes in the plasma membrane. Deletion of RCH1 confers hypersensitivity to high concentrations of extracellular Ca2+ and tolerance to azoles and Li+, which phenocopies the deletion of CaPMC1 (C. albicans PMC1) encoding the vacuolar Ca2+ pump. Additive to CaPMC1 mutation, lack of RCH1 alone shows an increase in Ca2+ sensitivity, Ca2+ uptake and cytosolic Ca2+ level. The Ca2+ hypersensitivity is abolished by cyclosporin A and magnesium. In addition, deletion of RCH1 elevates the expression of CaUTR2 (C. albicans UTR2), a downstream target of the Ca2+/calcineurin signalling. Mutational and functional analysis indicates that the Rch1p TM8 domain, but not the TM9 and TM10 domains, are required for its protein stability, cellular functions and subcellular localization. Therefore Rch1p is a novel regulator of cytosolic Ca2+ homoeostasis, which expands the functional spectrum of the vertebrate SLC10 family.

scholarly journals Recycling and Conversion of Yeasts into Organic Nitrogen Sources for Wine Fermentation: Effects on Molecular and Sensory Attributes

Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 313
Author(s):  
Paula Rojas ◽  
Daniel Lopez ◽  
Francisco Ibañez ◽  
Camila Urbina ◽  
Wendy Franco ◽  
...  
Keyword(s):  
Organic Nitrogen ◽  
Protein Hydrolysate ◽  
Principal Component ◽  
Nitrogen Sources ◽  
Isoamyl Alcohol ◽  
Positive Influence ◽  
Higher Alcohols ◽  
Fermentation Performance ◽  
High Concentrations ◽  
Significant Positive Influence

Organic nitrogen plays a significant role in the fermentation performance and production of esters and higher alcohols. This study assessed the use of yeast protein hydrolysate (YPH) as a nitrogen source for grape must fermentation. In this study, we prepared an enzymatic protein hydrolysate using yeasts recovered from a previous fermentation of wine. Three treatments were performed. DAP supplementation was used as a control, while two YPH treatments were used. Low (LDH) and high degrees of hydrolysis (HDH), 3.5% and 10%, respectively, were chosen. Gas chromatography and principal component analysis indicated a significant positive influence of YPH-supplementations on the production of esters and higher alcohols. Significantly high concentrations of 3-methyl-1-penthanol, isoamyl alcohol, isobutanol, and 2-phenylethanol were observed. Significant odorant activity was obtained for 3-methyl-1-pentanol and ethyl-2-hexenoate. The use of YPH as nitrogen supplementation is justified as a recycling yeasts technique by the increase in volatile compounds.

scholarly journals EFFECTS OF HUMAN PLATELET LYSATES WITHOUT ADDITIONAL GROWTH FACTORS ON THE PROTEIN PROFILES OF HUMAN UMBILICAL VEIN ENDOTHELIAL CELL CULTURE MEDIA

2017 ◽  
Vol 10 (17) ◽  
pp. 54
Author(s):  
Hazriani Ra ◽  
Lisa Amir ◽  
Ratna Farida
Keyword(s):  
Culture Media ◽  
Umbilical Vein ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Test Results ◽  
Control Groups ◽  
Cell Culture Media ◽  
Endothelial Cell Culture ◽  
Fetal Bovine

Objectives: The objective of this study is to evaluate the effect of HPLs with no additional GFs on the HUVEC protein profile. Methods: HUVEC cultures were examined in groups as follows: Fetal bovine serum (FBS), 2%-HPL with a GF, and 2%- and 5%-HPL without a GF which were analyzed with a sodium dodecyl sulfate-polyacrylamide gel electrophoresis test. Results: The intensity, thickness, and molecular weight of HUVEC band proteins cultured without a GF were not significantly different compared to the control groups (FBS or HPL with a GF). Conclusions: No difference was found in the HUVEC protein profile after they were cultured with FBS and HPLs, with or without GFs.

Binding of mycoplasmas to solid phase adsorbents

2005 ◽  
Vol 53 (3) ◽  
pp. 299-307 ◽  
Author(s):  
Susan Szathmáry ◽  
Nandani Rajapakse ◽  
Ibolya Székely ◽  
E. Pitlik ◽  
Judit Bíró ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Adhesion Molecules ◽  
Bovine Serum ◽  
Solid Phase ◽  
Fetal Bovine Serum ◽  
Vaccine Production ◽  
Fetal Bovine ◽  
Phosphate Buffered Saline ◽  
Lipid Structures

The capture of mycoplasmas (M. hominis, M. buccale, M. fermentans, M. bovis, M. synoviae, M. gallisepticum and M. arthritidis) based on lipid structures and adhesion molecules present in the mycoplasmal membrane was tested using different chromatographic resins (ActiClean Etox, ClarEtox, Heparin-Actigel, Sulfated Hiflow and SulfEtox). All of the resins efficiently reduced mycoplasma concentrations in Phosphate Buffered Saline (PBS) and in Fetal Bovine Serum (FBS) by 3-8 logs in a few minutes. This technology could be used for removing mycoplasmas from tissue culture components such as serum, and for concentrating mycoplasmas in vaccine production.

Parametric Effects of the Performance of Nanofiltration Membrane

2012 ◽  
Vol 248 ◽  
pp. 31-36
Author(s):  
Ayo Samuel Afolabi ◽  
Kazeem Oladele Sanusi ◽  
Ambali Saka Abdulkareem ◽  
Jaafar Bux Mohamed
Keyword(s):  
Membrane Filtration ◽  
Applied Pressure ◽  
Positive Influence ◽  
Excessive Force ◽  
Test Rig ◽  
Parametric Effects ◽  
Pressure Concentration ◽  
High Concentrations ◽  
Feed Temperature ◽  
Significant Positive Influence

The effects of pressure, concentration and feed temperature using a high pressure membrane filtration test rig capable of handling nanofiltration and reverse osmosis separation was investigated. The experimental results indicated that the applied pressure across the membrane has a significant positive influence on the flux produced and increasing the pressure will rupture the membrane if excessive force or pressure is applied. The high concentrations of solutes/molecules inhibit the flow of fluid through the membrane consequently lower fluxes and improved separation is and subsequently all variables, including their interactions, examined was found to be significant.

scholarly journals Decreased Sarcopenia in Aged Females with Young Ovary Transplants was Preserved in Mice that Received Germ Cell-Depleted Young Ovaries

2019 ◽  
Vol 8 (1) ◽  
pp. 40 ◽  
Author(s):  
Tracy L. Habermehl ◽  
Jeffrey B. Mason
Keyword(s):  
Body Composition ◽  
Germ Cell ◽  
Positive Influence ◽  
Test Results ◽  
Control Groups ◽  
Health Span ◽  
Treatment Groups ◽  
Significant Difference ◽  
Aging Health ◽  
Grip Test

Previously, transplantation of young, cycling, ovaries increased life and health span in post-reproductive female mice. The current study addressed the influence of ovarian germ cells in the improvement in health by performing transplantations of young, germ cell-depleted ovaries. The purpose of this study is to further the understanding of reproductive influences on aging health. Control mice were grouped by age. Treatment mice were age-matched and received either germ cell depleted ovaries or germ cell containing ovaries at 400 days of age. All groups underwent health span assays until sacrifice (treatment and age-matched control groups were between 680 and 700 days). Body composition results displayed an improvement of body composition in both treatment groups, compared to the controls, but no significant difference between the germ cell-depleted or germ cell-containing groups. Grip test results showed no improvement in musculoskeletal endurance and no change to mild loss of grip strength with both transplant groups compared to control groups. The research presented here suggests that reproductive status has a positive influence in post-reproductive health. A portion of this influence may be germ cell independent.

scholarly journals Reduced serum methods for contact-based coculture of human dermal fibroblasts and epidermal keratinocytes

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 347-355
Author(s):  
Snehal Kadam ◽  
Madhusoodhanan Vandana ◽  
Karishma S Kaushik
Keyword(s):  
Wound Closure ◽  
Fetal Bovine Serum ◽  
Cell Types ◽  
High Serum ◽  
Dermal Fibroblasts ◽  
Epidermal Keratinocytes ◽  
Human Dermal Fibroblasts ◽  
Fetal Bovine ◽  
High Concentrations ◽  
Serum Free Conditions

Direct contact-based coculture of human dermal fibroblasts and epidermal keratinocytes has been a long-standing and challenging issue owing to different serum and growth factor requirements of the two cell types. Existing protocols employ high serum concentrations (up to 10% fetal bovine serum), complex feeder systems and a range of supplemental factors. These approaches are technically demanding and labor intensive, and pose scientific and ethical limitations associated with the high concentrations of animal serum. On the other hand, serum-free conditions often fail to support the proliferation of one or both cell types when they are cultured together. We have developed two reduced serum approaches (1–2% serum) that support the contact-based coculture of human dermal fibroblasts and immortalized keratinocytes and enable the study of cell migration and wound closure.

scholarly journals Trypanosoma cruzi strains: behavior after passage into authoctonous or foreign species of triatomine (biological and biochemical patterns)

1996 ◽  
Vol 38 (1) ◽  
pp. 23-28 ◽  
Author(s):  
Juracy B Magalhães ◽  
Sonia G Andrade ◽  
Italo Sherlock
Keyword(s):  
Trypanosoma Cruzi ◽  
Positive Influence ◽  
Biological Behavior ◽  
Type Ii ◽  
Biological Characterization ◽  
Control Groups ◽  
Swiss Mice ◽  
Foreign Species

The behavior of T. cruzi strains from S. Felipe - BA (19 SF, 21 SF and 22 SF) classified as Type II Zymodeme 2, was investigated after passage through the authoctonous (P. megistus) and foreign vectors (T. infestans and R. prolixus). For each strain Swiss mice were infected: I - with blood forms (control); II - with metacyclic forms (MF) from P. megistus; III - with MF from T. infestans; IV - with MF from R. prolixus. Inocula: MF from the three species of triatomine, 60 to 120 days after feeding in infected mice, adjusted to 10 4. Biological behavior in mice (parasitemia, morphology, mortality, virulence and pathogenicity) after passage through triatomine was compared with data from the same strain in control mice. Isoenzymic electrophoresis (ASAT, ALAT, PGM, GPI) were also performed after culture into Warren medium. The three strains maintained the isoenzyme profiles (zymodeme 2), in the control groups and after passages through different species of triatomine. Biological characterization disclosed Type II strains patterns for all groups. An increased virulence was observed with the 22 SF strain isolated from P. megistus and T. infestans and higher levels of parasitemia and predominance of slender forms in mice inoculated with the 19 SF and 21 SF from these same species. Results indicate that the passage through the two species T. infestans and P. megistus had a positive influence on the virulence of the regional strains of S. Felipe, regardless of being autocthonous (P. megistus) or foreign to the area (T. infestans).

scholarly journals Assessment of TiO2 photoactivity on the lead removal: kinetic and mechanistic processing

2017 ◽  
Vol 75 (11) ◽  
pp. 2508-2519 ◽  
Author(s):  
Anca Peter ◽  
Anca Mihaly-Cozmuta ◽  
Camelia Nicula ◽  
Leonard Mihaly-Cozmuta
Keyword(s):  
Citric Acid ◽  
Uv Light ◽  
Removal Rate ◽  
Catalyst Particle ◽  
Positive Influence ◽  
Lead Removal ◽  
Starting Solution ◽  
Pseudo Second Order ◽  
High Concentrations ◽  
The Impact

Removal of lead ions from aqueous solutions, in the presence and in absence of commercial TiO2, under UV-light was studied. The influence of catalyst mass, concentration of Pb(II) ions and of citric acid in the starting solution as well as the impact of illumination conditions on the removal rate were also investigated. The results were fitted on the pseudo-first order, pseudo-second order and Elovich kinetic models, Weber-Morris intraparticle and liquid film diffusion models, in order to establish the photoreduction mechanism. The Pb removal rate increased in time by a combined linear-exponential mechanism. The catalyst had a positive influence on the removal rate at the start of the photoreduction. At a low concentration of Pb(II) ions (20 mg/L), the amount of photoreduced Pb(II) ions was proportional to the concentration of the citric acid solution, but at high concentrations (240 mg/L), the correlation was inversely proportional. The rate limiting steps in the removal of lead were both the photoreduction on the TiO2 surface and the diffusion through the film surrounding the catalyst particle. The lead removal was significantly influenced by the mobility and by the concentration of the species in solution.

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