scholarly journals Micronucleus Assay in Environmental Biomonitoring

2020 ◽  
Vol 64 (2) ◽  
pp. 20-28
Author(s):  
V. Michalová ◽  
M. Galdíková ◽  
B. Holečková ◽  
S. Koleničová ◽  
V. Schwarzbacherová
Keyword(s):  
Blood Cells ◽  
Crop Yields ◽  
Simultaneous Detection ◽  
Micronucleus Assay ◽  
Human Medicine ◽  
Farm Animals ◽  
Proliferation Index ◽  
Cytotoxic Effects ◽  
Chemical Pesticide ◽  
Chromosomal Aberration Test

AbstractNowadays many chemicals are widely used in agriculture to ensure high crop yields or in veterinary/human medicine to cure diseases. After their improper usage they may contaminate the environment, persist in it and adversely affect both the target and/or the non-target organisms. One of the ways to detect the occurrence of chemicals in the environment is to assess their impact on aquatic and farm animals; both are directly or indirectly exposed via their feed and water. The micronucleus assay is a standardly used cytogenetic test for the simultaneous detection of clastogenic and aneugenic agents. Additionally, cytotoxic effects are also assessed by analysing the proliferation changes using the cytokinesis-blocked proliferation index. The occurrence of micronuclei is analysed in many types of cells like the peripheral blood cells, bone marrow or cell lines according to standards for micronuclei detection. The analysis of published results has shown that the micronucleus assay is, together with the chromosomal aberration test, one of the most often used test in genotoxicity assessment. Its results have contributed to reassessing the use of multiple chemicals available on the market. Moreover, it is a compulsory test before approving the chemical/ pesticide for the market.

scholarly journals A Tool Derived from the Vicia faba Micronucleus Assay, to Assess Genotoxicity, Cytotoxicity or Biostimulation of Novel Compounds Used in Agriculture

Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 321
Author(s):  
Perrine Klein ◽  
Lorelei Chauvey ◽  
Jean Kallerhoff ◽  
Eric Pinelli ◽  
Marie Morard ◽  
...  
Keyword(s):  
Vicia Faba ◽  
Maleic Hydrazide ◽  
Lead Nitrate ◽  
Micronucleus Assay ◽  
Growth Potential ◽  
Protective Effects ◽  
Genotoxic Effects ◽  
Cytotoxic Effects ◽  
Conventional Agriculture ◽  
Rapid Tests

The increased use of biostimulants in conventional agriculture and organic farming requires the implementation of rapid tests to determine their effectiveness in enhancing plant growth and protection against abiotic stresses. However, their innocuity to plant health has rarely been demonstrated. We used the Vicia faba Micronucleus Assay, as described by the standard AFNOR EN ISO 29200(2020-05) to reveal biostimulant, genotoxic and cytotoxic effects of four commercialized wood-based products by comparing mitotic indices and micronucleus frequencies with respect to the controls. Neither genotoxicity, as measured by micronucleus frequency (MN), nor cytotoxicity, assessed by Mitotic index counts, was observed. Additionally, one of these stimulants (BHS®) conferred protective effects against contaminants (maleic hydrazide or lead nitrate). We describe that plotting micronuclei frequency against mitotic indices allows discrimination between cytotoxic/genotoxic effects from growth levels. Vicia faba experiments were successfully transposed to other agronomical important crops such as corn and sunflower. This technique can be valuable to industrials, to assess growth, potential cytoxicity and genotoxicity effects of any new biostimulant or organic.

scholarly journals Satureja subspicata and S. horvatii Extracts Induce Overexpression of the BCl-2 Family of Anti-apoptotic Genes and Reduce Micronuclei Frequency in Mice

2018 ◽  
Vol 13 (6) ◽  
pp. 1934578X1801300
Author(s):  
Jasmina Čakar ◽  
Naida Kadrić Lojo ◽  
Anja Haverić ◽  
Maida Hadžić ◽  
Lejla Lasić ◽  
...  
Keyword(s):  
Human Lymphocyte ◽  
Micronucleus Assay ◽  
Balkan Peninsula ◽  
Lymphocyte Culture ◽  
Cytotoxic Activities ◽  
Cytotoxic Effects ◽  
Apoptotic Genes ◽  
Apoptotic Activity

Satureja subspicata and S. horvatii are endemic species of the Balkan Peninsula and often used in traditional medicine in Bosnia and Herzegovina to treat different health conditions. We aimed to analyze the unevaluated apoptotic, genotoxic and cytotoxic effects of two Satureja species, as well as their content of phenolics that are mainly responsible for the plant's biological activity. Apoptotic and geno/cytotoxic activities of S. subspicata and S. horvatii were investigated in vitro in human lymphocyte culture and in vivo in mice. The content of the main phenolics in plant extracts was determined by ultra-high pressure liquid chromatography-MS-MS (UHPLC–MS/MS). Genotoxic and cytotoxic activities of Satureja extracts were evaluated in vitro by applying a cytokinesis-block micronucleus cytome assay in human lymphocyte culture and in vivo applying a mice reticulocytes micronucleus assay. SALSA RT-MLPA R011-C1 apoptosis assay was used for measuring the relative expression of 44 genes associated with the regulation of the apoptotic pathways in human lymphocyte cultures treated with different concentrations of two Satureja extracts. The first analysis of phenolic compounds in S. horvatii and S. subspicata determined by an UHPLC-MS/MS method revealed high levels of rosmarinic and caffeic acids. Minor genotoxic potential was determined in relation to the tested concentrations while no cytostatic and cytotoxic effects were revealed in vitro. However, when applied in concentrations of 200 mg/kg per os, aqueous extracts of two Satureja species significantly decreased frequency of reticulocytes micronuclei in treated mice against controls. Extracts of S. subspicata and S. horvatii in concentrations of 0.2 mg/mL, regardless of solvent used, downregulated pro-apoptotic and upregulated anti-apoptotic genes, showing anti-apoptotic activity. Our results indicate that the registered anti-genotoxic and anti-apoptotic activity is most likely related to the high level of phenolic acids (particularly rosmarinic and caffeic) in the tested extracts.

Use of a Multiplex PCR System for the Simultaneous Detection of Heat Labile Toxin I and Heat Stable Toxin II Genes of Enterotoxigenic Escherichia coli in Skim Milk and Porcine Stool

1998 ◽  
Vol 61 (2) ◽  
pp. 141-145 ◽  
Author(s):  
HAU-YANG TSEN ◽  
LIANG-ZHAO JIAN ◽  
WAN-RONG CHI
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Skim Milk ◽  
Pcr Primers ◽  
Enterotoxigenic Escherichia Coli ◽  
Farm Animals ◽  
Target Cells ◽  
Heat Stable ◽  
Heat Labile

Enterotoxigenic Escherichia coli (ETEC) strains which produce heat labile and/or heat stable toxins (LT and ST) may cause diarrhea in humans and farm animals. Using PCR primers specific for the LT I and ST II genes, a multiplex PCR system which allows detection of LT I- and ST II-producing ETEC strains was developed. When skim milk was used for a PCR assay, it was found that if target cells in the sample were precultured in MacConkey broth for 8 h prior to PCR as few as 100 cells per ml of the sample could be detected. Without the preculture step, 104 CFU of target cells per 0.2 g of porcine stool specimen were required to generate visible PCR products. The multiplex PCR System can be used for rapid testing of fecal specimens, food and possibly environmental samples for the presence of ETEC strains.

Resistance of Paroxysmal Nocturnal Hemoglobinuria Cells to the Glycosylphosphatidylinositol-Binding Toxin Aerolysin

Blood ◽  
1999 ◽  
Vol 93 (5) ◽  
pp. 1749-1756 ◽  
Author(s):  
Robert A. Brodsky ◽  
Galina L. Mukhina ◽  
Kim L. Nelson ◽  
Tracy S. Lawrence ◽  
Richard J. Jones ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Cells ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Bacterial Pathogen ◽  
Target Cells ◽  
Type Ii ◽  
Cytotoxic Effects ◽  
Standard Flow Cytometry ◽  
Kinetics Of ◽  
Cell Disorder

Paroxysmal nocturnal hemoglobinuria (PNH) is a clonal stem cell disorder caused by a somatic mutation of the PIGA gene. The product of this gene is required for the biosynthesis of glycosylphosphatidylinositol (GPI) anchors; therefore, the phenotypic hallmark of PNH cells is an absence or marked deficiency of all GPI-anchored proteins. Aerolysin is a toxin secreted by the bacterial pathogen Aeromonas hydrophila and is capable of killing target cells by forming channels in their membranes after binding to GPI-anchored receptors. We found that PNH blood cells (erythrocytes, lymphocytes, and granulocytes), but not blood cells from normals or other hematologic disorders, are resistant to the cytotoxic effects of aerolysin. The percentage of lysis of PNH cells after aerolysin exposure paralleled the percentage of CD59+ cells in the samples measured by flow cytometry. The kinetics of red blood cell lysis correlated with the type of PNH erythrocytes. PNH type III cells were completely resistant to aerolysin, whereas PNH type II cells displayed intermediate sensitivity. Importantly, the use of aerolysin allowed us to detect PNH populations that could not be detected by standard flow cytometry. Resistance of PNH cells to aerolysin allows for a simple, inexpensive assay for PNH that is sensitive and specific. Aerolysin should also be useful in studying PNH biology.

Cellular Toxicity of Inorganic Hydroxide Nanoparticles

2007 ◽  
Vol 7 (11) ◽  
pp. 4017-4020 ◽  
Author(s):  
Soo-Jin Choi ◽  
Jae-Min Oh ◽  
Taeun Park ◽  
Jin-Ho Choy
Keyword(s):  
Blood Cells ◽  
Membrane Damage ◽  
Cell Types ◽  
Carcinoma Cells ◽  
Chemical Compositions ◽  
Cytotoxic Effects ◽  
Cellular Toxicity ◽  
Anionic Clays ◽  
Hemolytic Effect ◽  
Double Hydroxides

Layered double hydroxides (LDHs), anionic clays, have attracted increasing interest as nanovehicles for delivering genes, drugs, and bio-active molecules into cells. However, no attempts have been made to evaluate the potential undesirable effects of LDH nanoparticles. The cytotoxicity of LDHs with different chemical compositions (ZnAl- and MgAl-LDH) was systematically evaluated in various cell types, such as human normal cells, carcinoma cells, and red blood cells, by measuring cell viability, cell proliferation, membrane damage, and hemolytic effect. No significant cytotoxic effects could be seen in both cases, but ZnAl-LDH was determined to be slightly more toxic than MgAl-LDH in terms of membrane damage and hemolysis induction. It is, therefore, expected that LDHs could be promising candidates for novel inorganic drug delivery carriers.

scholarly journals Assessment of the genotoxic potential of temephos

2020 ◽  
Vol 35 (3) ◽  
pp. 183-191
Author(s):  
Hayal Cobanoglu ◽  
Akin Cayir
Keyword(s):  
Sister Chromatid ◽  
Sister Chromatid Exchange ◽  
Human Peripheral Blood ◽  
Organophosphorus Pesticides ◽  
Peripheral Blood Lymphocytes ◽  
Proliferation Index ◽  
Genotoxic Effects ◽  
Cytotoxic Effects ◽  
Genotoxic Potential ◽  
Human Peripheral Lymphocytes

Genotoxic effects of pesticides are of great concern for public health due to the fact that they are widely used for both domestic and industrial purposes. Temephos is a member of organophosphorus pesticides, which is the most widely used group of chemicals against both agricultural and domestic insects. We therefore aimed in the present study to investigate the genotoxic and cytotoxic effects of temephos on human peripheral blood lymphocytes, using the cytokinesis-block micronucleus (CBMN) and sister chromatid exchange assays. The results showed that micronucleus (MN) frequency increased at concentrations of 50 and 75 ?g/ml although it was not found statically significant (p>0.05). We found that sister chromatid exchange (SCE) values at concentrations of 50 and 75 ?g/ ml were significantly higher than those obtained for the control (p<0.01). We also analyzed associations between temephos exposure and mitotic index (MI), proliferation index (PI), and cell blocked proliferation index (CBPI). There was no significant change in these values at the tested concentrations (p>0.05). It can be concluded that temephos was not cytotoxic at concentrations of 25, 50 and 75 ?g/ml. However, it may have a genotoxic potential in human peripheral lymphocytes.

scholarly journals Micronucleus Formation Induced by Glyphosate and Glyphosate-Based Herbicides in Human Peripheral White Blood Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Károly Nagy ◽  
Roba Argaw Tessema ◽  
István Szász ◽  
Tamara Smeirat ◽  
Alaa Al Rajo ◽  
...  
Keyword(s):  
Active Ingredient ◽  
Blood Cells ◽  
White Blood Cells ◽  
Micronucleus Assay ◽  
Metabolic Activation ◽  
Active Principle ◽  
Cell Viability Assay ◽  
Viability Assay ◽  
Peripheral White Blood Cells ◽  
Synergistic Toxicity

Glyphosate is the most commonly used herbicide around the world, which led to its accumulation in the environment and consequent ubiquitous human exposure. Glyphosate is marketed in numerous glyphosate-based herbicide formulations (GBHs) that include co-formulants to enhance herbicidal effect of the active ingredient, but are declared as inert substances. However, these other ingredients can have biologic activity on their own and may interact with the glyphosate in synergistic toxicity. In this study, we focused to compare the cytogenetic effect of the active ingredient glyphosate and three marketed GBHs (Roundup Mega, Fozat 480, and Glyfos) by investigating cytotoxicity with fluorescent co-labeling and WST-1 cell viability assay as well as genotoxicity with cytokinesis block micronucleus assay in isolated human mononuclear white blood cells. Glyphosate had no notable cytotoxic activity over the tested concentration range (0–10,000 μM), whereas all the selected GBHs induced significant cell death from 1,000 μM regardless of metabolic activation (S9). Micronucleus (MN) formation induced by glyphosate and its formulations at sub-cytotoxic concentrations (0–100 μM) exhibited a diverse pattern. Glyphosate caused statistically significant increase of MN frequency at the highest concentration (100 μM) after 20-h exposure. Contrarily, Roundup Mega exerted a significant genotoxic effect at 100 μM both after 4- and 20-h exposures; moreover, Glyfos and Fozat 480 also resulted in a statistically significant increase of MN frequency from the concentration of 10 μM after 4-h and 20-h treatment, respectively. The presence of S9 had no effect on MN formation induced by either glyphosate or GBHs. The differences observed in the cytotoxic and genotoxic pattern between the active principle and formulations confirm the previous concept that the presence of co-formulants in the formulations or the interaction of them with the active ingredient is responsible for the increased toxicity of herbicide products, and draw attention to the fact that GBHs are still currently in use, the toxicity of which rivals that of POEA-containing formulations (e.g., Glyfos) already banned in Europe. Hence, it is advisable to subject them to further comprehensive toxicological screening to assess the true health risks of exposed individuals, and to reconsider their free availability to any users.

scholarly journals Influence of proinflamatory macrophages (M-1) on proliferative activity and PPAR-γ expression in neoplastic rat hepatocytes in vitro

2019 ◽  
Vol 75 (05) ◽  
pp. 6220-2019
Author(s):  
MARTA WÓJCIK ◽  
URSZULA KOSIOR-KORZECKA ◽  
MICHAŁ PLEWIK ◽  
RYSZARD BOBOWIEC
Keyword(s):  
Blood Cells ◽  
Proliferative Activity ◽  
Gradient Centrifugation ◽  
Liver Perfusion ◽  
Negative Relationship ◽  
Rat Hepatocytes ◽  
Neoplastic Cell ◽  
Proliferation Index ◽  
Ppar Γ

We sought to analyse the proliferative activity and PPARγ expression in neoplastic and non-neoplastic rat hepatocytes, exposed to immunologically trained macrophages M1 (Mf-M1). Ten-week-old female Wistar rats were divided into two groups: I - control (n=5) and II neoplstic (n=5). To induce HCC in the neoplastic group, genotoxic diethylnitrosamine (DEN) was administered after a partial hepatectomy (PH). Hepatocytes were isolated by liver perfusion method and the mononuclear blood cells were isolated using Lymphoprep density-gradient centrifugation. After differentiation, blood cells were treated with barley-derived β-glucan (BBG) (10 μg/ml). Adhered heaptocytes and Mf were cultured in 3D Quasi-Vivo System during 24 h. Then, hepatic proliferation and PPAR γ expression were analysed after 72 h and the 1st ,the 2nd and the 3rd week of incubation. The proliferation index of control hepatocytes ranged between 0.41±0.04 – 0.43±0.03 after 72 h and after the 3rd week of incubation respectively. Exposure of these cells to Mf resulted in marked increase of IP after the 1st, the 2nd and the 3rd week of incubation. When hepatocytes were influenced by proinflammatory Mf- M1, their proliferation was maintained at control stage. DEN-obtained hepatocytes, not influenced by macrophages, exhibit enhanced proliferation. When these cells where co-cultured with Mf-M1, marked (P≤0.05) inhibition of cell proliferation was observed. In such condition, a high negative relationship (r= -0.93) between proliferative activity of the hepatocytes and the PPARγ concentration was observed. We conclude that immunologically trained macrophages M1, are capable to activation of PPARγ in rat neoplastic hepatocytes derived from experimentally induced HCC. In turn intensified expression of PPARγ may inhibited proliferation of neoplastic cell in vitro.

scholarly journals Clinical aspects of demodecosis in veterinary and human medicine

2017 ◽  
Vol 73 (5) ◽  
pp. 265-271 ◽  
Author(s):  
Dawid Jańczak ◽  
Anna Ruszczak ◽  
Ilona Kaszak ◽  
Elżbieta Gołąb ◽  
Karolina Barszcz
Keyword(s):  
Host Selection ◽  
Skin Disease ◽  
Hair Follicles ◽  
Human Medicine ◽  
Farm Animals ◽  
Clinical Aspects ◽  
Sebaceous Glands ◽  
Chronic Conjunctivitis

Mites from the genus Demodex are ectoparasites of many mammals, including humans. There are over 100 Demodex species, which demonstrate strong specificity in host selection [Table 1]. The mites are common in humans. It has been estimated that up to 60% of adults may be infected, but in most cases no symptoms of the disease are present. Demodex multiplication inside sebaceous glands and hair follicles can lead to skin disease in both humans and animals. In humans, the main problem is ocular demodecosis, which can cause chronic conjunctivitis and blepharitis. In this paper, we present the biology and epidemiology of Demodex species in humans as well as in domestic and farm animals. Characteristic lesions, diagnostics and treatment of demodecosis are also described

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