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1,027 result(s) for "EXPERIMENTATION IN VITRO"
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Antioxidant and antimicrobial activities of beet root pomace extracts
We described the in vitro antioxidant and antimicrobial activities of ethanol, acetone, and water extracts of beet root pomace. Total contents of phenolics (316.30-564.50 mg GAE/g of dry extract), flavonoids (316.30-564.50 mg RE/g of dry extract), betacyanins (18.78-24.18 mg/g of dry extract), and betaxanthins (11.19-22.90 mg/g of dry extract) after solid-phase extraction were determined spectrophotometrically. The antioxidant activity was determined by measuring the reducing power and DPPH scavenging activity by spectrometric method, and hydroxyl and superoxide anion radical scavenging activity by ESR spectroscopy. In general, the reducing power of all the beet root pomace extracts increased with increasing concentrations. The DPPH-free radical scavenging activity of the extracts, expressed as EC50, ranged from 0.133 mg/mL to 0.275 mg/mL. Significant correlation was observed between all phytochemical components and scavenging activity. Ethanol extract (0.5 mg/mL) completely eliminated hydroxyl radical which had been generated in Fenton system, while the same concentration of this extract scavenged 75% of superoxide anion radicals. In antibacterial tests, Staphylococcus aureus and Bacillus cereus showed higher susceptibility than Escherichia coli and Pseudomonas aeruginosa.
Evaluation of various plant extracts against the early blight disease of tomato plants under greenhouse and field conditions
The antimicrobial activity of six plant extracts from Ocimum basilicum, Azadirachta indica, Eucalyptus chamadulonsis, Datura stramonium, Nerium oleander, and Allium sativum was tested for controlling Alternaria solani in vitro and in vivo. In in vitro study the leaf extracts of D. stramonium, A. indica, and A. sativum at 5% concentration caused the highest reduction of mycelial growth of A. solani (44.4, 43.3 and 42.2%, respectively), while O. basilicum at 1% and 5% concentration and N. oleander at 5% concentration caused the lowest inhibition of mycelial growth of the pathogen. In greenhouse experiments the highest reduction of disease severity was achieved by the extracts of A. sativum at 5% concentration and D. stramonium at 1% and 5% concentration. The greatest reduction of disease severity was achieved by A. sativum at 5% concentration and the smallest reduction was obtained when tomato plants were treated with O. basilicum at 1% and 5% concentration (46.1 and 45.2 %, respectively). D. stramonium and A. sativum at 5% concentration increased the fruit yield by 76.2% and 66.7% compared to the infected control. All treatments with plant extracts significantly reduced the early blight disease as well as increased the yield of tomato compared to the infected control under field conditions.
Identification and abiotic stress response of a glutamine synthetase gene (AccGS) from the Asiatic honeybee, Apis cerana cerana (Hymenoptera: Apidae)
Glutamine synthetase (GS) is an essential detoxification enzyme that plays an important role in stress responses; however, little information regarding the function of this enzyme in hymenopteran insects is available. In the present study, we isolated and characterized the gene encoding GS in the Asiatic honeybee, Apis cerana cerana. Multiple alignments and a phylogenetic analysis of GS sequences showed that AccGS belongs to the GSII superfamily and clusters with invertebrate GSs. Real-time quantitative PCR data demonstrated that AccGS is expressed at all developmental stages and in all tissues, with the highest expression observed in the sixth larval instar and in the brain. Moreover, AccGS expression is highly regulated by environmental stress, including xenobiotic, temperature, and ultraviolet light stresses. A disc diffusion assay showed that the recombinant AccGS protein confers resistance to mercuric chloride (HgCl2) stress in E. coli. This suggests that AccGS may play multiple roles in early development and in environmental stress responses.
Nematicidal activity of a biopolymer chitosan at different molecular weights against root-knot nematode, Meloidogyne incognita
The nematicidal activity of four molecular weights (2.27 x 10**5, 3.60 x 10**5, 5.97 x 10**5, and 9.47 x 10**5 g/mol) of a biopolymer chitosan was assayed against the root-knot nematode, Meloidogyne incognita, in vitro and in pot experiments. In laboratory assays, the nematode mortality was significantly influenced by exposure times and chitosan molecular weight. Low molecular weight chitosan (2.27 x 10**5 g/mol) was the most effective in killing the nematode with EC50 of 283.47 and 124.90 mg/L after 24 and 48 h of treatment, respectively. In a greenhouse bioassay, all the compounds mixed in soil at one- and five-fold concentrations of the LC50 value significantly reduced population, egg mass, and root galling of tomato seedlings compared with the untreated control. In general, the nematicidal activity of these compounds was increased dramatically with a decrease in the molecular weight. The results suggest that the chitosan at low molecular weight may serve as a natural nematicide.
In vitro simulated digestion on the biostability of Hibiscus cannabinus L. seed extract
We investigate the biostability of phenolic acids from a kenaf (Hibiscus cannabinus L.) seed extract using an in vitro model simulating the physicochemical (pH, temperature and bile salts) and biological (gastric and pancreatic enzymes) gastrointestinal conditions. Some of the antioxidants in the kenaf seed extract were not relatively stable in the intestinal phase of the gastrointestinal tracts. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity and total phenolic content (TPC) assays displayed similar trends as the biostability of phenolic acids, which decreased during the digestion process. The overall percentage loss of selected phenolic acids was 8.4–49.4% in the intestinal phase. For the overall in vitro digestion system, significant correlations between phenolic acids, total phenolic content (TPC) and antioxidant activity (DPPH) were obtained in all digestion phases with the exception of the gastric phase.
Adipose stem cell combined with plasma-based implant bone tissue differentiation in vitro and in a horse with a phalanx digitalis distalis fracture: a case report
A horse suffering from an undetected hoof bone fracture was diagnosed three weeks after injury. The formation of callus tissue was detected at the fracture site. Standard orthopaedic screw application was augmented by a novel method, a combination of stem cells and plasma components. For experimental therapy, fat tissue and blood samples were collected from the patient to isolate stem cells and plasma proteins. The obtained and characterised mesenchymal stem cell population was applied to the wound area, together with an implant prepared from plasma, wrapped over the orthopaedic screw. Additionally, cells with implant were differentiated in vitro into bone tissue, to evaluate if cells could successfully produce extracellular matrix in such material. Three weeks after application, the hoof was significantly regenerated. The bone was completely rebuilt after three months. The in vitro experiment also gave positive results, with completely differentiated cells after three weeks. Our data show that enriching the standard orthopaedic material with mesenchymal stem cells adds therapeutic value to the treatment of refractory bone fractures.
Prion-inducing domain 2-114 of yeast Sup35 protein transforms in vitro into amyloid-like filaments
The yeast non-Mendelian genetic factor [PSI], which enhances the efficiency of tRNA-mediated non-sense suppression in Saccharomyces cerevisiae, is thought to be an abnormal cellular isoform of the Sup35 protein. Genetic studies have established that the N-terminal part of the Sup35 protein is sufficient for the genesis as well as the maintenance of [PSI]. Here we demonstrate that the N-terminal polypeptide fragment consisting of residues 2-114 of Sup35p, Sup35pN, spontaneously aggregates to form thin filaments in vitro. The filaments show a beta-sheet-type circular dichroism spectrum, exhibit increased protease resistance, and show amyloid-like optical properties. It is further shown that filament growth in freshly prepared Sup35pN solutions can be induced by seeding with a dilute suspension of preformed filaments. These results suggest that the abnormal cellular isoform of Sup35p is an amyloid-like aggregate and further indicate that seeding might be responsible for the maintenance of the [PSI] element in vivo
Ca2+-dependent protein kinases and stress signal transduction in plants
Stress responses in plants involve changes in the transcription of specific genes. The constitutively active mutants of two related Ca2+-dependent protein kinases (CDPK1 and CDPK1a) activate a stress-inducible promoter, bypassing stress signals. Six other plant protein kinases, including two distinct CDPKs, fail to mimic this stress signaling. The activation is abolished by a CDPK1 mutation in the kinase domain and diminished by a constitutively active protein phosphatase 2C that is capable of blocking responses to the stress hormone abscisic acid. A variety of functions are mediated by different CDPKs. CDPK1 and CDPK1a may be positive regulators controlling stress signal transduction in plants.