Search results for "Assay"

Development of competitive enzyme-linked immunosorbent assays for boscalid determination in fruit juices

2012

Abstract Boscalid is a modern, broad-spectrum carboxamide pesticide highly efficient against most fungal diseases affecting valuable crops. In this study, a boscalid-mimicking derivative with a six-carbon spacer arm replacing the chlorine atom at the pyridine ring of the target molecule was synthesized and coupled to carrier proteins. Following rabbit immunization, antibodies against this agrochemical were obtained for the first time, and they were characterised in terms of affinity and specificity, tolerance to solvents, and robustness to changes in buffer pH and ionic strength, using two assay formats. Both of the optimised immunoassays showed limits of detection below 0.1 μg/L. Moreover,…

A Specific Radioimmunoassay for the Determination of Low Quantities of Indole-3-acetic Acid in Spruce Needles of Healthy and Damaged Trees

1988

Summary The aim of the present investigation was to develop a radioimmunoassay for the quantification of indole-3-acetic acid in needles of Picea abies in damaged and phenotypically healthy trees. Phenols, lipophilic substances or compounds with cross-reactivity had to be separated from the extracts in several purification steps. Measurements were carried out between May and October 1986 on trees from two spruce plantations. Marked differences were found in IAA content in needles of healthy and damaged trees. In most cases the needles of the damaged trees contain lower endogenous IAA levels. The auxin levels also depended on the age of the tree, the year of needle formation, and the vegetat…

The Effect of Phosphinothricin (Glufosinate) on Glutathione Synthesis in Plants

1990

The inhibitory effect of DL-phosphinothricin (glufosinate) on glutathione synthesis was studied in vivo and in vitro. The influence of phosphinothricin on γ-glutamylcysteine synthetase was compared with the already known effects of l-buthionine sulfoximine and l-methionine sulfoximine. The results showed that phosphinothricin and buthionine sulfoximine are inhibitors of γ-glutamylcysteine synthetase of plants. With both substances the enzyme was inhibited by 50 % at a concentration of 7 . 10−4M (pI50 = 3.15). Methionine sulfoximine reduced the enzyme activity by 50% at 5 . 10−2 M (pI50 = 1.30). It is discussed that the target enzyme of phosphinothricin is the glutamine synthetase whereas th…

Neuroprotective and Mitoprotective Effects of Lemon IntegroPectin on SH-SY5Y Cells

2021

AbstractLemon IntegroPectin obtained via hydrodynamic cavitation of organic lemon processing waste in water shows significant neuroprotective activity in vitro, as first reported in this study investigating the effects of both lemon IntegroPectin and commercial citrus pectin on cell viability, cell morphology, reactive oxygen species (ROS) production and mitochondria perturbation induced by treatment of neuronal SH-SY5Y human cells with H2O2. Mediated by ROS including H2O2 and its derivatives, oxidative stress alters numerous cellular processes, including mitochondrial regulation and cell signaling, propagating cellular injury that leads to incurable neurodegenerative diseases. These result…

Glutathione-dependent defence system and monooxygenase enzyme activities in Arctic charr Salvelinus alpinus (L.) exposed to ozone

2000

Abstract One of the major obstacles of the increasing usage of ozone in aquaculture is the lack of relevant risk assessment in culture conditions. Before the apparent advantages of ozonation can be utilised efficiently, the safety margins and biological basis of ozone toxicity should be assessed. In this research, 1-year-old Arctic charr ( Salvelinus alpinus (L.)) were exposed to an ozone concentration high enough to inactivate Aeromonas sp. bacteria in freshwater, but too low to be directly lethal to the fish themselves. The effects of ozone exposure on the activity of glutathione-dependent antioxidant enzymes and monooxygenase reactions were studied in blood and in liver. The fish were ac…

2004

β-oxidation of long and very long chain fatty acyl-CoA derivatives occurs in peroxisomes, which are ubiquitous subcellular organelles of eukaryotic cells. This pathway releases acetyl-CoA as precursor for several key molecules such as cholesterol. Numerous enzymes participating to cholesterol and fatty acids biosynthesis pathways are co-localized in peroxisomes and some of their encoding genes are known as targets of the NFY transcriptional regulator. However, until now no interaction between NFY transcription factor and genes encoding peroxisomal β-oxidation has been reported. This work studied the interactions between NFY factor with the rat gene promoters of two enzymes of the fatty acid…

Inhibition of herpesvirus DNA synthesis by 9-beta-D-arabinofuranosyladenine in cellular and cell-free systems.

1977

9-beta-D-Arabinofuranosyladenine 5'-triphosphate (ara-ATP) is an inhibitor both of DNA polymerase-alpha and -beta from noninfected rabbit kidney cells and of the DNA-dependent DNA polymerase induced by herpes simplex virus Type 1 (strain IES). The studies were performed with partially purified enzymes, and each of the different polymerase preparations contained only one DNA-dependent DNA polymerase species. These enzymes were inhibited in a competitive manner. The HSV-induced DNA-dependent DNA polymerase was 39-fold more sensitive to ara-ATP than was cellular DNA polymerase-beta and 116-fold more sensitive than cellular DNA polymerase-alpha. The affinity of the HSV-induced enzyme for ara-AT…

Kinetics in a Double Antibody Radioimmunoassay (RIA): Diffussion Control

2012

Competitive protein binding radioimmunoassay (CPB-RIA) is a principal method for quantifying serum C Peptide concentration. The accuracy of this method is critically dependent on factors that influence the reaction between anti-C Peptide antibody (P) with 125I-C Peptide (M). We studied the influence of initial concentration of M, ionic strength, and viscosity on the reaction between M and P. A kinetic model for the the reaction between. Such model adjusts satisfac- torily to the results. Bi-exponential and irreversible kinetics is determined. The results of the viscosity analysis show clear negative influence on the direct reaction rate. The ionic strength shows scarce influence on equilibr…

Immediate early response of the marine sponge Suberites domuncula to heat stress: Reduction of trehalose and glutathione concentrations and glutathio…

1997

The marine sponge Suberites domuncula was used to identify early markers for thermal stress. Cubes from sponges have been kept for 30 min at 31°C (10 °C higher than the ambient temperature). After this treatment the sponge cubes were kept again at 21°C. To demonstrate that the animals reacted to the elevated temperature, the expression of heat shock protein (HSP) was determined. Using an antibody raised against HSP70, it was found by Western blotting that the animals specifically express a 45 kDa polypeptide after heat treatment. It was shown that even after 10 min of heat treatment the steady-state concentration of trehalose drops by 40% from a base level of 13 nmol/mg protein. The activit…

Characterization of the Proteinases Present in Germinating Seeds of Scots Pine, Pinus sylvestris

1978

Methods were developed to determine proteinase activity in germinating seeds of Scots pine. The assays were based on the liberation of TCA-soluble peptides from haemoglobin at pH 3.7 and from casein at pH 5.4 and pH 7.0; the reaction products were determined by the Lowry method. — Endosperms separated from seeds at the time of rapid storage protein mobilization (seedling length between 20 and 50 mm) showed high proteinase activities in all three assays. Experiments with different inhibitors suggested that at least four enzymes were involved. One of the enzymes resembled mammalian and microbial pepsin-like acid proteinases: the pH optimum was 3.7 and the enzyme was inhibited by pepstatin.—Th…