Genotoxic evaluation of newly synthesized iminothiazolidinones

2017 ◽  
Vol 33 (11) ◽  
pp. 811-820
Author(s):  
Fatma Tulay Tugcu ◽  
Kadir Turhan ◽  
Mehmet Karadayi ◽  
Medine Gulluce
Keyword(s):  
Seed Germination ◽  
Rapd Analysis ◽  
Mutagenic Activity ◽  
Random Amplified Polymorphic Dna ◽  
E Coli ◽  
Toxicological Effects ◽  
Actual Use ◽  
Genotoxic Compounds ◽  
Test Materials

The current study was designed to assess the potential toxicological effects of newly synthesized iminothiazolidinones by employing Ames Salmonella, Escherichia coli WP2, Zea mays seed germination, and random amplified polymorphic DNA (RAPD) assay systems. The bacterial tester strains S. typhimurium TA1535, TA1537, TA1538, TA98, TA100, and E. coli WP2 uvrA were chosen to test the direct gene mutation inducing capabilities of the test materials in prokaryotic systems and Z. mays seeds for determination of potential toxicological effects in eukaryotic systems. OPA-3 and OPA-6 primers were used in the RAPD analysis to determine genotoxic activities on the eukaryotic genomes. According to the results, none of the test materials showed significant mutagenic activity on the bacterial tester strains at the chosen concentrations. Additionally, none of the tested compounds showed inhibition of the germination of Z. mays seeds. In contrast, the RAPD analysis results were inconsistent with the bacterial reversion assays and the seed germination assay results. All test materials significantly changed the RAPD profiles for OPA-3; however, only compound 5 showed a significant change for OPA-6 when compared with the control groups. In conclusion, the newly synthesized iminothiazolidinone derivatives (C1–C5) were determined as potentially genotoxic compounds and they should be checked with multiple toxicology test systems before further studies to determine their actual use.

Differentiation of faecal Escherichia coli from human and animal sources by random amplified polymorphic DNA-PCR (RAPD-PCR)

2004 ◽  
Vol 50 (1) ◽  
pp. 193-198 ◽  
Author(s):  
D. Venieri ◽  
A. Vantarakis ◽  
G. Komninou ◽  
M. Papapetropoulou
Keyword(s):  
Escherichia Coli ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Hierarchical Cluster ◽  
Group Method ◽  
Randomly Amplified Polymorphic Dna ◽  
Arbitrary Primers ◽  
E Coli ◽  
Rapd Pcr ◽  
Dna Pcr

In this study the assessment of randomly amplified polymorphic DNA (RAPD) analysis was established as a molecular epidemiological tool. RAPD analysis was performed to differentiate faecal Escherichia coli isolates from human and animal sources. E. coli strains (128) were isolated from human and animal faeces (from cattle and sheep). Genomic DNA was extracted and randomly amplified polymorphic DNA-PCR (RAPD-PCR) fingerprinting was performed. Seven arbitrary primers were tested with a view to discriminating between E. coli isolates from humans and E. coli isolates from animals. RAPD profiles were analysed with hierarchical cluster analysis using an unweighted pair group method. RAPD profiles obtained with three of the tested primers (1247, 1290 and 1254) established a distinct differentiation between E. coli isolates from humans and E. coli from animals. Low levels of misclassification and high levels of specificity make RAPD a sensitive, efficient and reliable means of distinguishing closely related strains.

scholarly journals RAPD Analysis for Determination of Components in Herbal Medicine

2007 ◽  
Vol 4 (s1) ◽  
pp. 21-23 ◽  
Author(s):  
V. M. Shinde ◽  
K. Dhalwal ◽  
K. R. Mahadik ◽  
K. S. Joshi ◽  
B. K. Patwardhan
Keyword(s):  
Quality Control ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Herbal Medicines ◽  
Tinospora Cordifolia ◽  
Herbal Preparation ◽  
Emblica Officinalis ◽  
Dried Fruit ◽  
Herbal Prescription

In this study, the RAPD (Random Amplified Polymorphic DNA) technique was employed for determination of the components in an Ayurvedic herbal prescription,Rasayana Churna. One-hundred-and-twenty decamer oligonucleotide primers were screened in the RAPD analysis to identify three Ayurvedic medicines, dried stem ofTinospora cordifolia, dried fruit ofEmblica officinalisand dried fruit ofTribulus terestris, the Ayurvedic prescription. Primer OPC-6 simultaneously generated three distinct amplicons, each specific to one component. The marker with 600 bp is specific toTinospora cordifolia; the marker 500 bp is specific toEmblica officinalisand the remaining marker >1000 bp was present inTribulus terestris. Presence of three herbal medicines was determined when RAPD reaction with OPC-6 was performed. The technique was proved to contribute to the identification of components in Ayurvedic herbal preparation and thus helping to serve as a complementary tool for quality control.

scholarly journals Characterization of typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the classical O55 serogroup by RAPD analysis

1999 ◽  
Vol 30 (4) ◽  
pp. 365-368 ◽  
Author(s):  
Dennys M. Girão ◽  
Sílvia Y. Bando ◽  
Valéria Brígido de C. Girão ◽  
Carlos A. Moreira-Filho ◽  
Sérgio Eduardo L. Fracalanzza ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Epidemiological Studies ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Typical Epec ◽  
Rapd Method ◽  
Atypical Strains

The genetic diversity of 41 typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the serogroup O55 was analyzed by using the random amplified polymorphic DNA (RAPD) method. All typical EPEC O55 strains were grouped in two clusters (A and C) and belonged to the serotype O55:H6, while cluster B included all atypical strains, which were of the serotype O55:H7. The three groups also included non-motile strains. RAPD may be a useful method for epidemiological studies on E. coli O55 infection.

scholarly journals 011 RAPD ANALYSIS IDENTIFIES GENETIC VARIATION AND RELATEDNESS WITHIN THE CRANBERRY VARIETY MCFARLIN

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 429b-429
Author(s):  
Richard Novy ◽  
Nicholi Vorsa ◽  
Kim Patten
Keyword(s):  
Cluster Analysis ◽  
Genetic Variation ◽  
Genetic Basis ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Washington State ◽  
Genetic Component ◽  
Morphological Descriptors ◽  
Related Individuals

The cranberry cultivar `HcFarlin', selected from a natural bog in Massachusetts in 1874, has become the most widely grown cultivar in the Northwestern U.S.A. Washington state growers have noted variable productivity among `McFarlin' bogs. The determination of whether there is a genetic basis for the variability has been made difficult by a paucity of reliable morphological descriptors in cranberry. A random amplified polymorphic DNA (RAPD) analysis of 45 clones sampled from 12 WA `McFarlin' bogs identified 17 unique RAPD profiles. Cluster analysis identified 7 groups having various numbers of distinct, but related individuals. Eight clones were found to have RAPD profiles identical to the cultivar `Howes' indicating varietal misclassification had occurred in some bogs. One group of clones that originated from bogs classified as “Good” or “True” Mcfarlin' by growers had RAPD profiles similar to those of representatives from WI and MA `Mcfarlin' bogs. RAPD analysis has shown that `McFarlin' is represented by several genotypes, suggesting that the observed variability in production may have a genetic component.

Concentration determination of chromatin in unstained resin-embedded sections by means of Z-contrast in STEM

1990 ◽  
Vol 48 (2) ◽  
pp. 186-187
Author(s):  
M. Haider ◽  
B. Bohrmann
Keyword(s):  
Energy Loss ◽  
Freeze Substitution ◽  
Biological Macromolecules ◽  
Local Concentration ◽  
E Coli ◽  
Concentration Determination ◽  
Phosphorous Content ◽  
Z Contrast ◽  
Electron Energy Loss

The technique of Z-contrast in STEM offers the possibility to determine the local concentration of macromolecules like lipids, proteins or DNA. Contrast formation depends on the atomic composition of the particular structure. In the case of DNA, its phosphorous content discriminates it from other biological macromolecules. In our studies, sections of E. coli, the dinoflagellate Amphidinium carterae and Euglena spec. cells were used which were obtained by cryofixation followed by freeze-substitution into acetone with 3% glutaraldehyde. The samples were then embedded either in Lowicryl HM20 at low temperature or in Epon at high temperature. Sections were coated on both sides with 30Å carbon.The DF- and the inelastic image have been recorded simultaneously with a Cryo-STEM. This Cryo-STEM is equipped with a highly dispersive Electron Energy Loss Spectrometer. With this instrument pure Z-contrast can be achieved either with a Filtered DF-image divided by the inelastic image or, as is used in this paper, by dividing the conventional DF-image by an inelastic image which has been recorded with an inelastic detector whose response is dependent on the total energy loss of the inelastically scattered electrons.

Co-expression of recombinant single chain variable fragment recognizing blood antigen fused with sumo and chaperones in Escherichia coli

2018 ◽  
Vol 40 (4) ◽  
Author(s):  
Dang Thi Ngoc Ha ◽  
Le Thi Thu Hong ◽  
Truong Nam Hai
Keyword(s):  
Recombinant Antibody ◽  
Blood Type ◽  
Soluble Form ◽  
Supernatant Fraction ◽  
Blood Group Antigens ◽  
Single Chain ◽  
E Coli ◽  
Recombinant Scfv ◽  
Single Chain Variable Fragments

Single chain variable fragments (scFv) have widely been used in research, diagnosis and treatment, but the scFv is considered as difficult protein for expression in E. coli. In previous studies, we expressed a construction of recombinant single chain variable fragments again antigen specific for blood type A (antiA-scFv) individually or fused with Trx or SUMO. However, soluble fraction was low abandant and only approximately 40% when fused with Trx, the other cases were expressed in form of inclusion body. Therefore, it was difficult for purification, refolding and activity assesment. In thispaper, we demonstrated a suitable construction for soluble production of antiA-scFv fused with SUMO (SM/antiA-scFv) in presence of chaparones. Under fermentation with 0.1 mM IPTG at 20oC, the SM/antiA-scFv was entirely expressed in soluble form. Importantly, after cleavage from SUMO with SUMOprotease, antiA-scFv was still maintained in the supernatant fraction. Therefore, it can help ensure bioactivity and is useful for purification process. To the best of our knowledge, this is the first report showing soluble recombinant scFv fused with SUMO in presence of chaperone for determination of blood group antigens. Thus, this result facilitates the optimal study of soluble expression, purification and bioactivity determination of the antiA-scFv recombinant antibody. 

DETERMINATION OF AFTER-RIPENING PERIOD AND HORMONAL RESPONSE OF OROBANCHE SOLMSII C.D. CLARKE SEEDS

1970 ◽  
Vol 11 (1) ◽  
Author(s):  
A. Bista ◽  
G. B. Khattri ◽  
B. D. Acharya ◽  
S. C. Srivastava
Keyword(s):  
Growth Hormone ◽  
Seed Germination ◽  
Key Words ◽  
Incubation Period ◽  
Seed Set ◽  
Summer Season ◽  
Conditioning Period ◽  
Root Parasite ◽  
One Year

To find out the ability of Orobanche seeds to germinate immediately after seed set, seeds were germinated periodically at an interval of three months for one year in GR24. Some Orobanche seeds were capable of germination immediately after seed set but most required about nine months as after ripening or incubation period to be able to germinate. The phenomenon of after ripening in Orobanche seeds could be taken as an ecological measure to dormant over following unfavorable wet summer season. The growth hormone studies on Orobanche seed germination have shown that GA3 at a concentration of 100 ppm substantially enhanced seed germination when applied during pre-conditioning period. NAA showed some stimulatory effect at 0.5 - 1.0 ppm when applied during post-conditioning period but the hormone if applied during pre-conditioning period inhibited the germination. Kinetin failed to stimulate the germination at all the concentrations tested. Key words: Germination, root-parasite, hormone. Ecoprint Vol.11(1) 2004.

Transport of escherichia coli through soil to groundwater traced by randomly amplified polymorphic DNA (RAPD)

1997 ◽  
Vol 35 (11-12) ◽  
pp. 351-357 ◽  
Author(s):  
R. Rothmaier ◽  
A. Weidenmann ◽  
K. Botzenhart
Keyword(s):  
Negative Impact ◽  
Random Amplified Polymorphic Dna ◽  
Coli Strain ◽  
Soil Samples ◽  
Test Field ◽  
Liquid Manure ◽  
E Coli ◽  
Rapd Pattern ◽  
Geological Situation ◽  
Different Strains

Isolates (50) of E. coli obtained from liquid manure (20 bovine, 20 porcine) were genotyped using random amplified polymorphic DNA (RAPD). Typing revealed 9 and 14 different strains in bovine and porcine liquid manure respectively with no strains in common. One porcine strain, showing a simple RAPD pattern, was subcultured and spread on a test field (1.5l/m2 at 1010 cfu/l) in a drinking water protection zone with loamy to sandy sediments in the Donauried area, Baden-Wurttemberg. Soil samples and groundwaters were collected at monthly intervals October 1994 – June 1995 during which 114 E. coli isolates were recovered. The first occurrence and maximum concentration of E. coli in soil samples taken from more than 20cm depth was in January 1995, declining rapidly with depth and time. All isolates from soil and only one from groundwater showed the RAPD pattern of the spread E. coli strain. The results could not demonstrate a severe negative impact of the spreading of liquid manure on the bacteriological quality of the groundwater in the given geological situation. The distinct strain patterns found in different kinds of liquid manure suggest that genotyping of E. coli by RAPD may be an adequate tool for tracing sources of faecal contamination.

Identification of N-Substituted Triazolo-azetidines as Novel Antibacterials using pDualrep2 HTS Platform

2019 ◽  
Vol 22 (5) ◽  
pp. 346-354
Author(s):  
Yan A. Ivanenkov ◽  
Renat S. Yamidanov ◽  
Ilya A. Osterman ◽  
Petr V. Sergiev ◽  
Vladimir A. Aladinskiy ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Large Scale ◽  
Underlying Mechanism ◽  
Luciferase Assay ◽  
Reporter System ◽  
E Coli ◽  
Starting Point ◽  
High Concentrations ◽  
Mic Value

Aim and Objective: Antibiotic resistance is a serious constraint to the development of new effective antibacterials. Therefore, the discovery of the new antibacterials remains one of the main challenges in modern medicinal chemistry. This study was undertaken to identify novel molecules with antibacterial activity. Materials and Methods: Using our unique double-reporter system, in-house large-scale HTS campaign was conducted for the identification of antibacterial potency of small-molecule compounds. The construction allows us to visually assess the underlying mechanism of action. After the initial HTS and rescreen procedure, luciferase assay, C14-test, determination of MIC value and PrestoBlue test were carried out. Results: HTS rounds and rescreen campaign have revealed the antibacterial activity of a series of Nsubstituted triazolo-azetidines and their isosteric derivatives that has not been reported previously. Primary hit-molecule demonstrated a MIC value of 12.5 µg/mL against E. coli Δ tolC with signs of translation blockage and no SOS-response. Translation inhibition (26%, luciferase assay) was achieved at high concentrations up to 160 µg/mL, while no activity was found using C14-test. The compound did not demonstrate cytotoxicity in the PrestoBlue assay against a panel of eukaryotic cells. Within a series of direct structural analogues bearing the same or bioisosteric scaffold, compound 2 was found to have an improved antibacterial potency (MIC=6.25 µg/mL) close to Erythromycin (MIC=2.5-5 µg/mL) against the same strain. In contrast to the parent hit, this compound was more active and selective, and provided a robust IP position. Conclusion: N-substituted triazolo-azetidine scaffold may be used as a versatile starting point for the development of novel active and selective antibacterial compounds.

scholarly journals Quantitative Determination of the Effects of He-Ne Laser Irradiation on Seed Thermodynamics, Germination Attributes and Metabolites of Safflower (Carthamus tinctorius L.) in Relation with the Activities of Germination Enzymes

Agronomy ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1411
Author(s):  
Rashida Perveen ◽  
Xiukang Wang ◽  
Yasir Jamil ◽  
Qasim Ali ◽  
Shafaqat Ali ◽  
...  
Keyword(s):  
Seed Germination ◽  
Energy Levels ◽  
Soluble Sugars ◽  
Building Blocks ◽  
Carthamus Tinctorius ◽  
Fast Decline ◽  
Seed Irradiation ◽  
Seed Reserves ◽  
Positive Correlations

The present investigation was undertaken to assess the effects of different doses (100, 300, and 500 mJ) of low power He–Ne laser (632.8 nm) irradiation on seed germination and thermodynamics attributes and activities of potential germinating enzymes in relation with changes in seed metabolites. He–Ne laser seed irradiation increased the amylase (Amy), protease (Pro) and glucosidase (Gluco) activities, with a significant improvement in seed thermodynamics and seed germination attributes. A fast increase was found in free fatty acids (FFA), free amino acids (FAA), chlorophyll (Chl), carotenoids (Car), total soluble sugars (TSS) and reducing sugars (RS) in laser treated seeds in parallel with fast decline in seed oil contents and total soluble proteins (TSP). Significant positive correlations were recorded in laser-induced enhanced seed energy levels, germination, activities of germination enzymes with levels of FAA, FFA, Chl, TSS and RS, but a negative correlation with the levels of TSP and oil. In conclusion, the seed treatment with 100 and 300 mJ He–Ne laser was more effective to improve the seed germination potential associated with an improvement in seed energy levels due to increased activities of germination enzymes due to the speedy breakdown of seed reserves to simple metabolites as building blocks.

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