scholarly journals Evaluation of viability indicators of bovine spermatozoa after exposure to silicon dimethylglycerolate using flow cytofluorimetry

2021 ◽  
Vol 209 (06) ◽  
pp. 53-60
Author(s):  
Alena Nakidkina ◽  
T. I. KUZMINA

Abstract. Silicon and its dioxide (silica) demonstrate good biological compatibility and a wide range of physical and chemical properties, depending on the production and processing method. In particular, silicon dimethylglycerolate (SDMG) has transmucous and transcutaneous drug conductivity, and, as a hydrogel, may be of interest for the oocytes and embryos cultivation medium structuring and/or media for cryopreservation/thawing of gametes. The aim of this study was to examine the effect of SDMG at concentrations of 0.2 % and 0.02 % on the transmembrane potential of mitochondria and cell viability of bovine spermatozoa. Methods. Sperm subpopulations were assessed for (non)viability indicators (disrupted transmembrane potential of mitochondria, externalization of phosphatidylserine and plasma membrane integrity loss) by flow cytometry with two sets of fluorescent probes. Mitochondrial transmembrane potential was measured using 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3))/ethidium bromide, and externalization of phosphatidylserine – using Annexin V-FITC/propidium iodide pair. The results of this work indicate that SDMG in concentrations of 0.2 % and 0.02 % does not affect the transmembrane mitochondrial potential, externalization of phosphatidylserine or necrotic processes in the population of bovine spermatozoa. The scientific novelty. The data is obtained for the first time on the absence of cytotoxicity of SDMG for male gametes. Together with the shown positive effect of this compound on the morphological parameters and the state of nuclear chromatin of porcine oocytes after intrafollicular vitrification, it should be concluded that silicon-containing glycerohydrogels are of interest as a component of sperm cryopreservation/thawing media.

Zygote ◽  
2012 ◽  
Vol 22 (3) ◽  
pp. 378-386 ◽  
Author(s):  
S.S.M. Faezah ◽  
F.M.Y Zuraina ◽  
J.H.F. Farah ◽  
O. Khairul ◽  
N.I. Hilwani ◽  
...  

SummaryCryopreservation is a technique used to preserve cells for long-time storage. It is widely used in agriculture to store male gametes in liquid nitrogen. The aim of this study was to determine the optimum thawing temperature and time for samples subjected to annexin V magnetic-activated cell sorting (AnMACS) as the sperm preparation technique. Pooled semen samples from three ejaculates were divided into two groups. The treatment group was subjected both to AnMACS and to being cryopreserved, whilst the control group was cryopreserved directly without MACS. Post-thaw analysis was carried out for samples thawed at either 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s or 80°C for 5 s. Sperm kinematics, viability and capacitation status were determined for samples subjected to all thawing temperatures described. Results showed that thawing at 37°C for 13 s for MACS-processed samples was a superior option compared with other thawing procedures; there was a significant difference in P < 0.05 values for curvilinear velocity (VCL μm/s) and sperm straightness (STR %) when samples were thawed at 40°C for 7 s, with fewer capacitated spermatozoa (P < 0.05) when samples were thawed at 37°C for 30 s, 40°C for 7 s or 60°C for 6 s. Hence, we can speculate that the use of AnMACS as the sperm preparation technique can somehow enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these cells has yet to be determined.


2019 ◽  
Vol 8 (2) ◽  
pp. 188-195 ◽  
Author(s):  
Sylwia Terpilowska ◽  
Andrzej K. Siwicki

The aim of this study was to examine the effect of chromium(iii), iron(iii), molybdenum(iii) and nickel(ii) and their combinations on the cell cycle and mitochondrial transmembrane potential (MTP) in BALB/3T3 and HepG2 cells.


2019 ◽  
Vol 19 (9) ◽  
pp. 1132-1140
Author(s):  
Heba A.E. Mohamed ◽  
Hossa F. Al-Shareef

Background: Quinolones are a significant group of nitrogen heterocyclic compounds that exist in therapeutic agents, alkaloids, and synthetic small molecules that have important biological activities. A wide range of quinolones have been used as antituberculosis, antibacterial, anti-malarial, antifungal, anticonvulsant, anticancer agents and urease inhibitors. Methods: Ethyl 3,3-disubstituted-2-cyano propionates containing hybride quinolones derivatives were synthesized by the reaction of 1-amino-7-hydroxy-4-methylquinolin-2(1H)-one and its dibromo derivative with α, β-unsaturated carbonyl in ethanol. Results: A novel series of hybrid 2-quinolone derivatives was designed and synthesized. The compounds structures were confirmed using different spectroscopic methods and elemental analysis. The cytotoxic activities of all the compounds were assessed against HepG2 cell line in comparison with doxorubicin as a standard drug. Conclusion: Most compounds revealed superior anti-proliferative activity than the standard. Compound 4b, is the most active compound (IC50 = 0.39mM) compared with doxorubicin (IC50 = 9.23mM). DNA flow cytometric analysis of compound 4b showed cell cycle arrest at G2/M phase with a concomitant increase of cells in apoptotic phase. Dual annexin-V/ propidium iodide staining assay of compound 4b revealed that the selected candidate increased the apoptosis of HepG-2 cells more than control.


2019 ◽  
Vol 19 (15) ◽  
pp. 1835-1845
Author(s):  
Ali Hassanzadeh ◽  
Adel Naimi ◽  
Majid F. Hagh ◽  
Raedeh Saraei ◽  
Faroogh Marofi ◽  
...  

Introduction: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L) is a member of the tumor necrosis factor (TNF) superfamily, which stimulates apoptosis in a wide range of cancer cells via binding to death receptors 4 and 5 (DR4/5). Nevertheless, TRAIL has noticeable anti-cancer abilities; some cancer cells acquire resistance to TRAIL, and consequently its potential for inducing apoptosis in target cells is strongly diminished. Acute lymphoblastic leukemia MOLT-4 cell line is one of the most resistant cells to TRAIL that developed resistance to TRAIL via different pathways. We used TRAIL plus kaempferol to eliminate resistance of the MOLT-4 cells to TRAIL. Material and Methods: First, IC50 for kaempferol (95 µM) was determined by using the MTT assay. Second, the viability of the MOLT-4 cells was assayed by FACS after Annexin V/PI staining, following treatment with TRAIL (50 and 100 nM) and kaempferol (95 µM) alone and together. Finally, the expression levels of the candidate genes involved in resistance to TRAIL were assayed by real-time PCR technique. Results: Kaempferol plus TRAIL induced apoptosis robustly in MOLT-4 cells at 12, 24 and 48 hours after treatment. Additionally, we found that kaempferol could inhibit expression of the c-FLIP, X-IAP, cIAP1/2, FGF-8 and VEGF-beta, and conversely augment expression of the DR4/5 in MOLT-4 cells. Conclusion: We suggest that co-treatment of MOLT-4 cells with TRAIL plus kaempferol is a practical and attractive approach to eliminate cancers’ resistance to TRAIL via inhibition of the intracellular anti-apoptotic proteins, upregulation of DR4/5 and also by suppression of the VEGF-beta (VEGFB) and FGF-8 expressions.


2021 ◽  
Vol 22 (15) ◽  
pp. 7906
Author(s):  
Alexey A. Komissarov ◽  
Maria A. Karaseva ◽  
Marina P. Roschina ◽  
Andrey V. Shubin ◽  
Nataliya A. Lunina ◽  
...  

Regulated cell death (RCD) is a fundamental process common to nearly all living beings and essential for the development and tissue homeostasis in animals and humans. A wide range of molecules can induce RCD, including a number of viral proteolytic enzymes. To date, numerous data indicate that picornaviral 3C proteases can induce RCD. In most reported cases, these proteases induce classical caspase-dependent apoptosis. In contrast, the human hepatitis A virus 3C protease (3Cpro) has recently been shown to cause caspase-independent cell death accompanied by previously undescribed features. Here, we expressed 3Cpro in HEK293, HeLa, and A549 human cell lines to characterize 3Cpro-induced cell death morphologically and biochemically using flow cytometry and fluorescence microscopy. We found that dead cells demonstrated necrosis-like morphological changes including permeabilization of the plasma membrane, loss of mitochondrial potential, as well as mitochondria and nuclei swelling. Additionally, we showed that 3Cpro-induced cell death was efficiently blocked by ferroptosis inhibitors and was accompanied by intense lipid peroxidation. Taken together, these results indicate that 3Cpro induces ferroptosis upon its individual expression in human cells. This is the first demonstration that a proteolytic enzyme can induce ferroptosis, the recently discovered and actively studied type of RCD.


Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1891
Author(s):  
Antonio Reina ◽  
Trung Dang-Bao ◽  
Itzel Guerrero-Ríos ◽  
Montserrat Gómez

Metal nanoparticles have been deeply studied in the last few decades due to their attractive physical and chemical properties, finding a wide range of applications in several fields. Among them, well-defined nano-structures can combine the main advantages of heterogeneous and homogenous catalysts. Especially, catalyzed multi-step processes for the production of added-value chemicals represent straightforward synthetic methodologies, including tandem and sequential reactions that avoid the purification of intermediate compounds. In particular, palladium- and copper-based nanocatalysts are often applied, becoming a current strategy in the sustainable synthesis of fine chemicals. The rational tailoring of nanosized materials involving both those immobilized on solid supports and liquid phases and their applications in organic synthesis are herein reviewed.


Radiocarbon ◽  
2001 ◽  
Vol 43 (2B) ◽  
pp. 731-742 ◽  
Author(s):  
D Lal ◽  
A J T Jull

Nuclear interactions of cosmic rays produce a number of stable and radioactive isotopes on the earth (Lai and Peters 1967). Two of these, 14C and 10Be, find applications as tracers in a wide variety of earth science problems by virtue of their special combination of attributes: 1) their source functions, 2) their half-lives, and 3) their chemical properties. The radioisotope, 14C (half-life = 5730 yr) produced in the earth's atmosphere was the first to be discovered (Anderson et al. 1947; Libby 1952). The next longer-lived isotope, also produced in the earth's atmosphere, 10Be (half-life = 1.5 myr) was discovered independently by two groups within a decade (Arnold 1956; Goel et al. 1957; Lal 1991a). Both the isotopes are produced efficiently in the earth's atmosphere, and also in solids on the earth's surface. Independently and jointly they serve as useful tracers for characterizing the evolutionary history of a wide range of materials and artifacts. Here, we specifically focus on the production of 14C in terrestrial solids, designated as in-situ-produced 14C (to differentiate it from atmospheric 14C, initially produced in the atmosphere). We also illustrate the application to several earth science problems. This is a relatively new area of investigations, using 14C as a tracer, which was made possible by the development of accelerator mass spectrometry (AMS). The availability of the in-situ 14C variety has enormously enhanced the overall scope of 14C as a tracer (singly or together with in-situ-produced 10Be), which eminently qualifies it as a unique tracer for studying earth sciences.


2007 ◽  
Vol 97 (2) ◽  
pp. 321-328 ◽  
Author(s):  
S. Aisling Aherne ◽  
Joseph P. Kerry ◽  
Nora M. O'Brien

Experimental evidence suggests that most herbs and spices possess a wide range of biological and pharmacological activities that may protect tissues against O2-induced damage. The objectives of the present study were: first, to determine the effects of plant extracts on the viability, membrane integrity, antioxidant status and DNA integrity of Caco-2 cells and second, to investigate the cytoprotective and genoprotective effects of these plant extracts against oxidative stress in Caco-2 cells. The plant extracts examined were rosemary (Rosmarinus officinalis L.), oregano (Origanum vulgare L.), sage (Salvia officinalis L.) and echinacea (Echinacea purpurea L.). Cell membrane integrity was assessed by the lactate dehydrogenase release assay. Viability was determined by the neutral red uptake assay (NRUA) and the concentration of compound that resulted in 50 % cell death (IC50) was calculated. Antioxidant status of the cells was assessed by measuring GSH content, catalase activity and superoxide dismutase activity. To examine their cytoprotective and genoprotective effects, Caco-2 cells were pre-treated with each plant extract for 24 h followed by exposure to H2O2. DNA damage was assessed by the comet assay and cell injury was determined by the NRUA. Rosemary was the most toxic (IC50 123 μg/ml) and echinacea the least toxic (IC50 1421 μg/ml). Sage was the only plant extract to affect the antioxidant status of the cells by increasing GSH content. Sage, oregano and rosemary protected against H2O2-induced DNA damage (olive tail moment and percentage tail DNA), whereas protection against H2O2-induced cytotoxicity was afforded by sage only.


1997 ◽  
Vol 496 ◽  
Author(s):  
David J. Derwin ◽  
Kim Kinoshita ◽  
Tri D. Tran ◽  
Peter Zaleski

AbstractSeveral types of carbonaceous materials from Superior Graphite Co. were investigated for lithium ion intercalation. These commercially available cokes, graphitized cokes and graphites have a wide range of physical and chemical properties. The coke materials were investigated in propylene carbonate based electrolytes and the graphitic materials were studied in ethylene carbonate / dimethyl solutions to prevent exfoliation. The reversible capacities of disordered cokes are below 230 mAh / g and those for many highly ordered synthetic (artificial) and natural graphites approached 372 mAh / g (LiC6). The irreversible capacity losses vary between 15 to as much as 200 % of reversible capacities for various types of carbon. Heat treated cokes with the average particle size of 10 microns showed marked improvements in reversible capacity for lithium intercalation. The electrochemical characteristics are correlated with data obtained from scanning electron microscopy (SEM), high resolution transmission electron microscopy (TAM), X - ray diffraction (XRD) and BET surface area analysis. The electrochemical performance, availability, cost and manufacturability of these commercial carbons will be discussed.


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