scholarly journals Development of a TaqMan qPCR protocol for detecting Acipenser ruthenus in the Volga headwaters from eDNA samples

2020 ◽  
Vol 12 (3) ◽  
pp. 395-397 ◽  
Author(s):  
Tamara Schenekar ◽  
Martin Schletterer ◽  
Steven J. Weiss
Keyword(s):  
Sensitivity And Specificity ◽  
In Silico ◽  
High Sensitivity ◽  
In Vitro Tests ◽  
Fish Diversity ◽  
Upper Volga ◽  
Acipenser Ruthenus ◽  
Taqman Qpcr

AbstractA recent eDNA-metabarcoding study assessing fish diversity in the Upper Volga catchment did not detect sterlet at any of the sampled stretches, despite recent sightings that suggest its presence. We designed a TaqMan qPCR protocol to test for sterlet in selected eDNA samples from that study. In-silico and in-vitro tests confirm the protocol’s high sensitivity and specificity to sturgeon taxa and potentially paddlefishes. Using this assay, sterlet were not detected in 26 eDNA samples from the Volga headwaters, agreeing with the metabarcoding results.

scholarly journals Predicting the bactericidal efficacy of solar disinfection (SODIS): from kinetic modeling of in vitro tests towards the in silico forecast of E. coli inactivation

2021 ◽  
pp. 130866
Author(s):  
Sofia Samoili ◽  
Giulio Farinelli ◽  
José Ángel Moreno-SanSegundo ◽  
Kevin G. McGuigan ◽  
Javier Marugán ◽  
...  
Keyword(s):  
Kinetic Modeling ◽  
In Silico ◽  
In Vitro Tests ◽  
Solar Disinfection ◽  
E Coli ◽  
Bactericidal Efficacy

scholarly journals Neo-antigens for the serological diagnosis of IgE-mediated drug allergic reactions to antibiotics cephalosporin, carbapenem and monobactam

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Edurne Peña-Mendizabal ◽  
Sergi Morais ◽  
Ángel Maquieira
Keyword(s):  
Drug Allergy ◽  
High Sensitivity ◽  
Specific Ige ◽  
In Vitro Tests ◽  
Allergic Reactions ◽  
Serum Samples ◽  
Lactam Ring ◽  
Specific Igg ◽  
Ige Mediated

Abstract New antigens deriving from -lloyl and -llanyl, major and minor determinants, respectively, were produced for β-lactam antibiotics cefuroxime, cefotaxime, ceftriaxone, meropenem and aztreonam. Twenty β-lactam antigens were produced using human serum albumin and histone H1 as carrier proteins. Antigens were tested by multiplex in vitro immunoassays and evaluated based on the detection of specific IgG and IgE in the serum samples. Both major and minor determinants were appropriate antigens for detecting specific anti-β-lactam IgG in immunised rabbit sera. In a cohort of 37 allergic patients, we observed that only the minor determinants (-llanyl antigens) were suitable for determining specific anti-β-lactam IgE antibodies with high sensitivity (< 0.01 IU/mL; 24 ng/L) and specificity (100%). These findings reveal that not only the haptenisation of β-lactam antibiotics renders improved molecular recognition events when the 4-member β-lactam ring remains unmodified, but also may contribute to develop promising minor antigens suitable for detecting specific IgE-mediated allergic reactions. This will facilitate the development of sensitive and selective multiplexed in vitro tests for drug-allergy diagnoses to antibiotics cephalosporin, carbapenem and monobactam.

scholarly journals An Integrated Decision-tree Testing Strategy for Repeat Dose Toxicity with Respect to the Requirements of the EU REACH Legislation

2008 ◽  
Vol 36 (1) ◽  
pp. 93-101 ◽  
Author(s):  
Christina Grindon ◽  
Robert Combes ◽  
Mark T.D. Cronin ◽  
David W. Roberts ◽  
John F. Garrod
Keyword(s):  
In Silico ◽  
New Technologies ◽  
In Vivo Studies ◽  
In Vitro Tests ◽  
Repeat Dose ◽  
The European Union ◽  
Testing Strategy ◽  
Repeat Dose Toxicity ◽  
Joint Research Project

This paper presents some results of a joint research project conducted by FRAME and Liverpool John Moores University, and sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity end-points associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for repeat dose (sub-acute, sub-chronic and chronic) toxicity testing. It reviews the limited number of in silico and in vitro tests available for this endpoint, and outlines new technologies which could be used in the future, e.g. the use of biomarkers and the ‘omics’ technologies. An integrated testing strategy is proposed, which makes use of as much non-animal data as possible, before any essential in vivo studies are performed. Although none of the non-animal tests are currently undergoing validation, their results could help to reduce the number of animals required for testing for repeat dose toxicity.

Research On Dental Amalgam: 1982-1986

1988 ◽  
Vol 2 (1) ◽  
pp. 71-82 ◽  
Author(s):  
D.B. Mahler
Keyword(s):  
Clinical Performance ◽  
High Sensitivity ◽  
Dental Amalgam ◽  
Performance Criterion ◽  
Dental Restoration ◽  
In Vitro Tests ◽  
Significant Finding ◽  
Amalgam Restorations

This paper is a five-year review of selected research papers on dental amalgam which were published during the years 1982 through 1986. Papers presented at scientific dental meetings are also included. During the past five years, clarification of amalgam metallurgy has been made, and a few innovative modifications have been recommended. The addition of palladium, indium, and selenium to dental amalgam has been suggested, but commercial applications have yet to be made. The mechanical property of creep has been studied more extensively, while the applicability of fracture toughness tests has been examined. More work has been done on the microleakage of amalgam restorations, with certain alloy factors showing an influence on this problem. Most in vitro investigations on the use of cavity varnish or resin films beneath amalgam restorations show reduced microleakage. Additionally, many electrochemical investigations were conducted. A most significant finding was that amalgam does not appear to break down in vivo as much as in vitro tests would indicate, the buffering action of saliva being protective in this regard. The marginal fracture evaluation of clinical amalgam restorations is still being used as a major clinical performance criterion, and measuring techniques for this failure mode have been improved. Measurements of metallic ion loss from amalgam were made with instruments of high sensitivity, but no evidence has been found to associate this loss with any disease entity. The rare presence of an allergy to mercury appears to be the only contra-indication for the use of amalgam as a dental restoration. Further research on dental amalgam can lead to improved clinical performance of this most useful restorative material.

Antimicrobial Evaluation, Molecular Docking and ADME Properties of Indole Amide Derivatives

2021 ◽  
Vol 18 ◽  
Author(s):  
Derya Doğanay ◽  
Sevval M. Özcan ◽  
Ahmet M. Şentürk ◽  
Süreyya Ölgen
Keyword(s):  
Molecular Docking ◽  
In Silico ◽  
Bacterial Infections ◽  
Antimicrobial Agents ◽  
Antimicrobial Properties ◽  
Antimicrobial Activities ◽  
In Vitro Tests ◽  
Lipinski’S Rule ◽  
Amide Derivatives

Background: Besides the viral infections, bacterial infections can cause serious and life-threatening complications and drug resistance is an important problem to fight bacterial infections. Therefore, it is important to discover novel antimicrobial agents to fight such infections. Objective: Several indole containing antimicrobial drug development studies have been reported in literature that provided strong evidences for good antimicrobial activities against a variety of microorganisms. Taken into consideration from these findings, antimicrobial properties of previously synthesized 16 indole amide derivatives were evaluated by in vitro tests against 14 different microorganisms, and also molecular docking and in silico prediction studies were used to identify structure-activity relationship of compounds. Methods: Antimicrobial activity of compounds was determined by disc diffusion and tube dilution methods. Molecular docking of compounds was studied to determine the relationship between the structure of compounds with DNA gyrase interactions of microorganisms by using the version of Autodock vina 4.2.6. Mol inspiration and Swiss ADME prediction online software programs were also used to identify drug-like properties of compounds. Results: The results showed that some compounds exhibited quite pronounced antibacterial and antifungal activities compared to reference drugs. These results were also supported by molecular docking studies and in silico ADME calculations presented that all tested compounds obey the Lipinski’s Rule of Five and are metabolized by CYP450 enzymes. Conclusion: It can be concluded that these results can be taken as reference in the development of new indole-based antimicrobial agents.

An Integrated Decision-tree Testing Strategy for Repeat Dose Toxicity with Respect to the Requirements of the EU REACH Legislation

2008 ◽  
Vol 36 (1_suppl) ◽  
pp. 139-147 ◽  
Author(s):  
Christina Grindon ◽  
Robert Combes ◽  
Mark T.D. Cronin ◽  
David W. Roberts ◽  
John F. Garrod
Keyword(s):  
In Silico ◽  
New Technologies ◽  
In Vivo Studies ◽  
In Vitro Tests ◽  
Repeat Dose ◽  
The European Union ◽  
Testing Strategy ◽  
Repeat Dose Toxicity ◽  
Joint Research Project

This paper presents some results of a joint research project conducted by FRAME and Liverpool John Moores University, and sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity end-points associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for repeat dose (sub-acute, sub-chronic and chronic) toxicity testing. It reviews the limited number of in silico and in vitro tests available for this endpoint, and outlines new technologies which could be used in the future, e.g. the use of biomarkers and the ‘omics’ technologies. An integrated testing strategy is proposed, which makes use of as much non-animal data as possible, before any essential in vivo studies are performed. Although none of the non-animal tests are currently undergoing validation, their results could help to reduce the number of animals required for testing for repeat dose toxicity.

scholarly journals Gamma-hexalactone flavoring causes DNA lesion and modulates cytokines secretion at non-cytotoxic concentrations

2019 ◽  
Vol 20 (S1) ◽  
Author(s):  
Luísa Zuravski ◽  
Taiane A. Escobar ◽  
Elizandra G. Schmitt ◽  
Queila D. F. Amaral ◽  
Fávero R. Paula ◽  
...  
Keyword(s):  
Comet Assay ◽  
In Silico ◽  
In Silico Analysis ◽  
Computational Prediction ◽  
Negative Control ◽  
Alcoholic Beverages ◽  
In Vitro Assays ◽  
In Vitro Tests ◽  
Dna Comet Assay

Abstract Background The γ-hexalactone is a flavoring agent for alcoholic beverages, teas, breads, dairy products, coffees, buttery products among others. It presents low molecular weight and exhibits sweet fruity aroma with nuances of nuts. As far as we know, both literature and government regulations have gaps regarding the safe use of the γ-hexalactone. In this context, the main objective of this work was to evaluate the effects of γ-hexalactone through in silico and in vitro approaches. Methods The in silico analysis was performed through four free online platforms (admetSAR, Osiris Property Explorer®, pkCSM platform and PreADMET) and consisted of comparative structural analysis with substances present in databases. The computational prediction was performed in the sense of complement and guide the in vitro tests. Regarding in vitro investigations, screening of cytotoxicity (assessed by cell proliferation and viability parameters) in lymphocytes exposed to γ-hexalactone for 72 h were carried out previously to determine non-cytotoxic concentrations. Following this screening, concentrations of 5.15, 0.515, and 0.0515 μM were selected for the study of the respective potentials: genotoxic (assessed by DNA comet assay), chromosomal mutation (analysis of micronucleus frequency) and immunomodulatory (cytokine quantification using ELISA immunoassay). The results of in vitro assays were compared by one-way analysis of variance (ANOVA), followed by Bonferroni’s post hoc test, conducted by statistic software. Results The platform PreADMET pointed out that γ-hexalactone is potentially mutagenic and carcinogenic. The comet assay data corroborate with these results demonstrating that γ-hexalactone at 5.15 μM caused lymphocytes DNA damage. In relation to cytokine secretion, the results indicate that lymphocytes were activated by γ-hexalactone at non-cytotoxic concentrations, involving an increase in the IL-1 levels in all tested concentrations, ranging from approximately 56 to 93%. The γ-hexalactone only at 5.15 μM induced increase in the levels of IL-6 (~ 60%), TNF-α (~ 68%) and IFN-γ (~ 29%), but decreased IL-10 (~ 46%) in comparison with the negative control (p < 0.05). No change was observed in total lymphocytes or in cell viability at the concentrations tested. Conclusions In summary, the γ-hexalactone demonstrated immunomodulatory and genotoxic effects at non-cytotoxic concentrations in healthy lymphocytes.

Designing of a multiepitopic antigen for anti-HBV antibody detection with high sensitivity and specificity: an in silico approach

2011 ◽  
Vol 44 (13) ◽  
pp. S313
Author(s):  
Saeed Khalili ◽  
Rasaee Mohamad Javad ◽  
Mousavi Seyed Latif ◽  
Jafar Amani ◽  
Abolfazl Jahangiri
Keyword(s):  
Sensitivity And Specificity ◽  
In Silico ◽  
High Sensitivity ◽  
Antibody Detection

scholarly journals Potential of Herbaspirillum and Azospirillum Consortium to Promote Growth of Perennial Ryegrass under Water Deficit

2021 ◽  
Vol 9 (1) ◽  
pp. 91
Author(s):  
Sandra Cortés-Patiño ◽  
Christian Vargas ◽  
Fagua Álvarez-Flórez ◽  
Ruth Bonilla ◽  
German Estrada-Bonilla
Keyword(s):  
Plant Growth ◽  
Water Deficit ◽  
Perennial Ryegrass ◽  
In Silico ◽  
Growth Promotion ◽  
Plant Colonization ◽  
In Vitro Tests ◽  
Plant Growth Promoting Bacteria ◽  
Growth Promoting

Plant growth-promoting bacteria (PGPB) can mitigate the effect of abiotic stresses on plant growth and development; however, the degree of plant response is host-specific. The present study aimed to assess the growth-promoting effect of Herbaspirillum (AP21, AP02), Azospirillum (D7), and Pseudomonas (N7) strains (single and co-inoculated) in perennial ryegrass plants subjected to drought. The plants were grown under controlled conditions and subjected to water deficit for 10 days. A significant increase of approximately 30% in dry biomass production was observed using three co-inoculation combinations (p < 0.01). Genomic analysis enabled the detection of representative genes associated with plant colonization and growth promotion. In vitro tests revealed that all the strains could produce indolic compounds and exopolysaccharides and suggested that they could promote plant growth via volatile organic compounds. Co-inoculations mostly decreased the in vitro-tested growth-promoting traits; however, the co-inoculation of Herbaspirillum sp. AP21 and Azospirillum brasilense D7 resulted in the highest indolic compound production (p < 0.05). Although the Azospirillum strain showed the highest potential in the in vitro and in silico tests, the plants responded better when PGPB were co-inoculated, demonstrating the importance of integrating in silico, in vitro, and in vivo assessment results when selecting PGPB to mitigate drought stress.

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