scholarly journals Adsorbents for the Sequestration of the Pimelea Toxin, Simplexin

Proceedings ◽  
2020 ◽  
Vol 36 (1) ◽  
pp. 90
Author(s):  
Gordon ◽  
Hungerford ◽  
Laycock ◽  
Ouwerkerk ◽  
Fletcher
Keyword(s):  
Rumen Fluid ◽  
Screening Method ◽  
Binding Activity ◽  
Rapid Screening ◽  
Sodium Bentonite ◽  
Arid Rangelands ◽  
Future Work ◽  
Calcium Bentonite ◽  
Binding Mechanisms

Pimelea poisoning affects cattle grazing arid rangelands of Australia, has no known remedy and significant outbreaks can cost the industry $50 million per annum. Poisoning is attributable to consumption of native Pimelea plants containing the toxin simplexin. Charcoal, bentonite and other adsorbents are currently used by the livestock industry to mitigate the effects of mycotoxins. The efficacy of such adsorbents to mitigate Pimelea poisoning warrants investigation. Through a series of in vitro experiments, different adsorbents were evaluated for their effectiveness to bind simplexin using a simple single concentration, dispersive adsorbent rapid screening method. Initial experiments were conducted in a rumen fluid based medium, with increasing quantities of each adsorbent: sodium bentonite (Trufeed®, Sibelco Australia), biochar (Nutralick®Australia) and Elitox® (Impextraco, Belgium). Data showed the unbound concentration of simplexin decreased with increasing quantities of each adsorbent tested. Sodium bentonite performed best, removing ~95% simplexin at 12 mg/mL. A second experiment using a single amount of adsorbent included two additional adsorbents: calcium bentonite (Bentonite Resources, Australia) and a synthetic adsorbent (Waters, USA). The concentration of simplexin remaining in the solution after 1 h, the amount able to be desorbed off the adsorbent-toxin matrix with replacement fresh fluid, and the amount remaining bound to the adsorbent were measured. All samples containing an adsorbent were statistically different compared to the blank (p < 0.05), indicating some binding activity. Future work will explore the binding mechanisms and behaviour of the toxin-adsorbent complex in the lower gastrointestinal tract.

scholarly journals Phytochemical Profiles, Physicochemical Analysis, and Biological Activity of Three Colchicum Species

2021 ◽  
Vol 16 (2) ◽  
Author(s):  
Ali Davoodi ◽  
Mohammad Azadbakht ◽  
Seyed Jalal Hosseinimehr ◽  
Saeed Emami ◽  
Masoud Azadbakht
Keyword(s):  
Physicochemical Properties ◽  
Large Scale ◽  
Protein Denaturation ◽  
Potassium Dichromate ◽  
Biological Effects ◽  
Screening Method ◽  
Rapid Screening ◽  
Total Phenolic ◽  
Anti Inflammatory

Background: Colchicum is a genus of the Colchicaceae family with various isolated compounds, especially tropolone alkaloids and flavonoids, which are used for osteoarthritis, gout, cancer, inflammatory diseases, jaundice, and sexual impotence in different societies. Objectives: The current study aimed to evaluate the phytochemical and physicochemical properties and anti-inflammatory activities of three Colchicum species. Methods: Total tropolone alkaloid, total phenolic/total tannin, and total flavonoid contents were determined using acidic potassium dichromate, Folin-Ciocalteu, and aluminum chloride methods, respectively. Moreover, the HPLC method was used for identification and quantitation purposes of tropolone alkaloids. Physicochemical properties of three Colchicum species, including macroscopic and organoleptic properties, solubility, foreign matter, ash values, and heavy metal contents, were evaluated. Besides, in vitro anti-inflammatory activities of the corms also were determined using the protein denaturation technique as a rapid screening method. Results: The highest levels of tropolone alkaloid, phenolic compounds, tannins, and flavonoids were observed in C. autumnale, C. speciosum, and C. robustum, respectively. HPLC analysis indicated the presence of colchicine, demecolcine, 2-demethyl colchicine, 3-demethyl colchicine, colchicoside, colchifoline, cornigerine, and N-deacetyl-N-formyl colchicine in these Colchicum species. The physicochemical properties of C. speciosum and C. robustum corms are appropriate compared to the standard Colchicum autumnale corm. Moreover, all Colchicum species exhibited high anti-inflammatory activities compared to standard drugs. Conclusions: This study demonstrated that the corm of the Colchicum species contained similar main compounds with different amounts, as well as appropriate physicochemical properties. Moreover, the valuable biological effects of these plants stimulate the cultivations on a large scale.

scholarly journals An in silico and in vitro pipeline for the rapid screening of helicase modulators

2020 ◽  
Vol 25 (1) ◽  
pp. e927
Author(s):  
Eleni Papakonstantinou ◽  
Flora Bacopoulou ◽  
Vasileios Megalooikonomou ◽  
Aspasia Efthimiadou ◽  
Dimitrios Vlachakis
Keyword(s):  
Enzyme Activity ◽  
High Throughput Screening ◽  
Screening Method ◽  
The Other ◽  
Rapid Screening ◽  
Helicase Activity ◽  
Luminescence Signal ◽  
Coated Plate ◽  
Biotin Molecule

To evaluate the potency of potential helicase modulators, we developed an assay of helicase enzyme activity. Using a DNA or RNA biotin labelled oligonucleotide and after the addition of a recombinant helicase, the nucleic acid unwinds, causing the emission of luminescence, which is quantified with a particular antibody. In our assay, one of the DNA oligos was biotinylated, while the other was labelled with digoxygenin (DIG), both in their 5’ termini. The biotin molecule immobilises the DNA duplex on a neutravidin-coated plate and the helicase activity is measured through the unwinding of DNA, due to ATP activation. The subsequent release of DIG-labelled oligos results in a luminescence signal measured with a chemiluminescence antibody. Our goal was to provide a high throughput screening method for potential helicase inhibitors. The method described in this paper has been demonstrated to be fast, easy and reproducible and doesn’t use radiochemicals.

scholarly journals An in silico and in vitro pipeline for the rapid screening of helicase modulators

2020 ◽  
Vol 25 ◽  
pp. e927
Author(s):  
Eleni Papakonstantinou ◽  
Flora Bacopoulou ◽  
Vasileios Megalooikonomou ◽  
Aspasia Efthimiadou ◽  
Dimitrios Vlachakis
Keyword(s):  
Enzyme Activity ◽  
High Throughput Screening ◽  
Screening Method ◽  
The Other ◽  
Rapid Screening ◽  
Helicase Activity ◽  
Luminescence Signal ◽  
Coated Plate ◽  
Biotin Molecule

To evaluate the potency of potential helicase modulators, we developed an assay of helicase enzyme activity. Using a DNA or RNA biotin labelled oligonucleotide and after the addition of a recombinant helicase, the nucleic acid unwinds, causing the emission of luminescence, which is quantified with a particular antibody. In our assay, one of the DNA oligos was biotinylated, while the other was labelled with digoxygenin (DIG), both in their 5’ termini. The biotin molecule immobilises the DNA duplex on a neutravidin-coated plate and the helicase activity is measured through the unwinding of DNA, due to ATP activation. The subsequent release of DIG-labelled oligos results in a luminescence signal measured with a chemiluminescence antibody. Our goal was to provide a high throughput screening method for potential helicase inhibitors. The method described in this paper has been demonstrated to be fast, easy and reproducible and doesn’t use radiochemicals.

scholarly journals Pengujian Nomor-nomor Harapan Padi Tahan Al dan pH Rendah Hasil Seleksi In Vitro dengan Kultur Hara

2016 ◽  
Vol 4 (1) ◽  
pp. 18 ◽  
Author(s):  
Ragapadmi Purnamaningsih ◽  
Ika Mariska
Keyword(s):  
Root Development ◽  
Root Length ◽  
In Vitro Selection ◽  
Acid Soil ◽  
Screening Method ◽  
Culture Method ◽  
Solution Culture ◽  
Low Ph ◽  
Rapid Screening

<p>Rice productivity in acid soil is very low because of low pH,<br />low availability of N, P, K, Ca, Mg, Mo, toxicity of Al and Mn.<br />Development of Al tolerant varieties could increase rice<br />productivity in acid soil. Somaclonal variation and in vitro<br />selection method can be used to develop new Al tolerance<br />varieties. A rapid screening method is needed to select a<br />large number of new genotypes or new inbred lines in plant<br />breeding, such as solution culture methods to evalu-ate Altolerant<br />rice. This methods was used to know the response<br />to Al in the seedling stage, root development, and pH<br />changing. In this experiment solution culture method was<br />used to evaluate the new genotypes derived from somaclonal<br />variation and in vitro selection methods. These new<br />genotypes have been tested the tolerance characteristic by<br />using AlCl36H2O at 6 concentrations (0, 100, 200, 300, 400,<br />and 500 ppm). Yoshida solution with two Al concentration<br />were used to tested these genotypes. Measurement of Al<br />tolerance was based on root development by using Relative<br />Root Length (RRL), the relativity of root length at 45 ppm and<br />0 ppm. Almost all of the genotypes have RRLs higher than<br />0.7, which means that there was a positive correlation<br />between the in vitro method and solution culture method. In<br />this experiment pH changes were not applicable to measure<br />the tolerance of the rice genotypes to Al and low pH.</p>

scholarly journals Use of Artificial Diets with Plant Material to evaluate Banana Cultivars for Resistance to Cosmopolites sordidus

2018 ◽  
Vol 18 (2) ◽  
pp. 103-109
Author(s):  
Elyeza Bakaze ◽  
Andrew Kiggundu ◽  
Wilberforce Tushemereirwe
Keyword(s):  
Control Strategies ◽  
Larval Growth ◽  
Screening Method ◽  
Rapid Screening ◽  
Preliminary Evaluation ◽  
Potential Candidate ◽  
Artificial Diets ◽  
Commercial Diet ◽  
Rapid Bioassay

Artificial diets rapidly establish the effectiveness of chemical-based control strategies. Diets permit preliminary evaluation of active compounds, study in-vitro larval growth cycles that are usually inaccessible and produce uniform large consistent numbers of insects as needed. With no known artificial diet, banana weevils, have always been reared on field-collected banana rhizome (corm). This study, therefore, developed and examined the effect of commercial diet recipes fortified with susceptible banana corm powder on weevil growth and development. Subsequently, corm powders from different banana cultivars were also evaluated for weevil performance. Successful laboratory rearing of the weevils to adult stage on diet was achieved in 48 days compared to 36 days in the natural banana stem. The difference in weevil larvae performance reared different corm powder, presented a novel screening method for banana genotypes. For example genotypes, Culcatta-4 (AA), Cavendish (AAA) and Kayinja (ABB) showed 0-35% of adult emergences compared to 65% in susceptible genotypes. The diet developed can be used to perform rapid bioassay experimentation to screen potential candidate proteins or molecules for a transgenic approach. It has also shown potential for rapid screening of genotypes for resistance. Keywords: Resistant banana, Laboratory weevil rearing, Resistance screening.

scholarly journals Development of a Robust Screening Method for Pathogenicity of Colletotrichum spp. on Strawberry Seedlings Enabling Forward Genetic Studies

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 845-851 ◽  
Author(s):  
Sigal Horowitz ◽  
Oded Yarden ◽  
Aida Zveibil ◽  
Stanley Freeman
Keyword(s):  
Large Scale ◽  
Developmental Stages ◽  
Screening Method ◽  
Rapid Screening ◽  
Disease Symptoms ◽  
Inoculation Techniques ◽  
Lower Temperature ◽  
Incubation Temperatures

Generation and screening for nonpathogenic mutants is a popular tool for identifying pathogenicity-related genes. Successful application of this technique for plant fungal pathosystems requires reliable and rapid screening procedures. This study reports on the development of a rapid in vitro bioassay enabling large-scale screening and isolation of nonpathogenic mutants of Colletotrichum gloeosporioides and C. acutatum on strawberry seedlings. Inoculation was carried out on strawberry seedlings at two different developmental stages: 12-week-old (young) and 15-week-old (older) seedlings. A comparison was made between two inoculation techniques, (i) foliar dip and (ii) root soak, at two incubation temperatures (19 and 25°C). Mortality of young seedlings was observed 4 days after inoculation with both species, reaching 50% within 10 days, using both techniques at 25°C. However, mortality of older seedlings was delayed by 4 days compared with that in the young seedlings when using the root-soak method. Disease development decreased in young and older seedlings at the lower temperature. This method also was reliable in determining pathogenicity of the cucurbit-specific C. magna that did not cause disease symptoms on strawberry by either inoculation method. The proposed method enabled screening of more than 980 restriction enzyme-mediated integration mutants resulting in a selection of five reduced-virulence isolates. Initial characterization of some of these mutants revealed large differences in germination and appressorial formation compared with pathogenic isolates.

An integrated screening method for evaluating the pulmonary toxicity of inhaled particulates: Role of microscopic techniques in assessing pulmonary macrophage clearance functions

1990 ◽  
Vol 48 (3) ◽  
pp. 826-827
Author(s):  
David B. Warheit ◽  
Lena Achinko ◽  
Mark A. Hartsky
Keyword(s):  
Bronchoalveolar Lavage ◽  
Pulmonary Toxicity ◽  
Screening Method ◽  
Pulmonary Response ◽  
Inhaled Particles ◽  
Cytochemical Analysis ◽  
Pulmonary Macrophages ◽  
Integrated Screening

There is a great need for the development of a rapid and reliable bioassay to evaluate the pulmonary toxicity of inhaled particles. A number of methods have been proposed, including lung clearance studies, bronchoalveolar lavage analysis, and in vitro cytotoxicity tests. These methods are often limited in scope inasmuch as they measure only one dimension of the pulmonary response to inhaled, instilled or incubated dusts. Accordingly, a comprehensive approach to lung toxicity studies has been developed.To validate the method, rats were exposed for 6 hours or 3 days to various concentrations of either aerosolized alpha quartz silica (Si) or carbonyl iron (CI) particles. Cells and fluids from groups of sham and dust-exposed animals were recovered by bronchoalveolar lavage (BAL). Alkaline phosphatase, LDH and protein values were measured in BAL fluids at several time points postexposure. Cells were counted and evaluated for viability, as well as differential and cytochemical analysis. In addition, pulmonary macrophages (PM) were cultured and studied for morphology, chemotaxis, and phagocytosis by scanning electron microscopy.

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