A Comparative In Vitro Evaluation of the Anti-Inflammatory Effects of a Tisochrysis lutea Extract and Fucoxanthin

In this study, we compared the effects of a Tisochrysis lutea (T. lutea) F&M-M36 methanolic extract with those of fucoxanthin (FX) at equivalent concentration, on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The T. lutea F&M-M36 methanolic extract contained 4.7 mg of FX and 6.22 mg of gallic acid equivalents of phenols per gram. HPLC analysis revealed the presence of simple phenolic acid derivatives. The T. lutea F&M-M36 extract exhibited a potent and concentration-dependent inhibitory activity against COX-2 dependent PGE2 production compared to FX alone. Compared to LPS, T. lutea F&M-M36 extract and FX reduced the expression of IL-6 and of Arg1 and enhanced that of IL-10 and of HO-1; T. lutea F&M-M36 extract also significantly abated the expression of NLRP3, enhanced mir-223 expression and reduced that of mir-146b, compared to LPS (p < 0.05). These findings indicate that T. lutea F&M-M36 methanolic extract has a peculiar anti-inflammatory activity against COX-2/PGE2 and NLRP3/mir-223 that might be attributable to the known anti-inflammatory effects of simple phenolic compounds found in the extract that may synergize with FX. Our data suggest that T. lutea F&M-M36 may serve as a source of anti-inflammatory compounds to be further evaluated in in vivo models of inflammation.     

Cassava common mosaic virus infection causes alterations in chloroplast ultrastructure,function, and carbohydrate metabolism of cassava plants

Cassava common mosaic virus (CsCMV) is a potexvirus that causes systemic infections in cassava plants, leading to chlorotic mosaic and producing significant yield losses. To date, the physiological alterations and the mechanism underlying biotic stress during the cassava–CsCMV compatible interaction remains unknown. In this study, we found that CsCMV infection adversely modified chloroplast structure and had functional effects on chloroplasts in source leaves during the course of viral infection. Extrusion of the chloroplast membrane with amoeboid-shaped appearance and disorganized grana stacks were observed in infected mesophyll cells. These alterations were associated with up to 35% reduction of relative chlorophyll content, and a decline of CO₂ fixation (13.5% and 24.2% at 90 and 210 days after planting, respectively). The effects of CsCMV infection on the performance index on absorption basis dropped up to 37%. The analysis of chlorophyll a fluorescence showed a progressive loss of both oxygen evolving complex activity and “connectivity” within the tripartite system (core antenna-LHCII-reaction centre). Here, we report the latter phenomenon for the first time in a viral infection. The oxidative stress process was observed in CsCMV-infected plants (20.8% reduction of antioxidant capacity with respect to noninfected plants). Other effects of the pathogen included reduction of starch and maltose content in source leaves, and a significant increase (24.7%) of the sucrose:starch ratio, which indicates an altered pattern of carbon allocation. Our results suggest that CsCMV induces chloroplast distortion associated with progressive chloroplast function loss and diversion of carbon flux in source leaf tissue, leading to the loss of cassava tuber yield.     

Utilization of Wastes from the Coke-Chemical Production and Sewage Sludge as Additives in the Brick-Clay

Current modern building technologies use a greater variety of building materials and elements of these. Lately, intensive work has been done on investigating possibilities for using different wastes as additives in the production of new ceramic building products. This demonstrates that the raw material base can be enlarged and some environmental problems solved.The present work investigates the possibility of combined utilization of sludge from the Sofia wastewater treatment plant and waste products from coke-chemical production as additives to brick clay. The most appropriate combinations and concentrations of the materials used, and the means of preparing and firing the brick masses are determined. The strength characteristics of the resulting products are studied. The results obtained are useful in showing possibilities for the solution of environmental problems.     

Soil‐less bioassays for early screening for resistance to imazapyr in sunflower (Helianthus annuus L.)

BACKGROUND: Rapid and efficient diagnostic tests for early screening of herbicide resistance are convenient alternatives to field screening methods. There is a need for a quick, reliable and cost‐effective method for rapid diagnosis of imidazolinone resistance in sunflower (Helianthus annuus L.). RESULTS: Two seed germination bioassays were developed. Seeds from three sunflower inbred lines differing in resistance to imidazolinones were germinated either on solid culture medium or placed in plastic pots filled with commercial perlite. After 8 days incubation under controlled conditions, both assays successfully distinguished susceptible genotype from the resistant and intermediate ones. The susceptible genotype showed arrested root growth at all herbicide treatments (root length < 1 cm). The resistant genotype developed a complete root system even when exposed to the highest dose of herbicide. However, no definite differences were observed for the intermediate and resistant genotypes with respect to root growth under the different herbicide treatments. CONCLUSION: The simple and rapid screening assays described in the present study were useful in discriminating imidazolinone resistance at the seedling stage. Therefore, these bioassays could be potential tools for early screening of imidazolinone resistance genes from large sunflower populations. Copyright © 2009 Society of Chemical Industry     

Measurement of nuclear DNA content of the genus <Emphasis Type="Italic">Trifolium</Emphasis> L. as a measure of genebank accession identity

The collection, identification and maintenance of genebank accessions of the genus Trifolium is a major task because of the large number of genera and their occasional morphological similarity. We investigated whether the measurement of nuclear DNA content can serve as an additional criterion for identification of mislabeled accessions. Relative nuclear DNA content was determined by flow cytometry measurements for a total of 151 genebank accessions of 23 Trifolium species with notable agronomical value. Among 23 species analyzed, 15 were found to possess a uniform relative nuclear DNA content, with intraspecific variability of the majority of analyzed species lower than 5%. Within six Trifolium species, 1–2 atypical accessions with outstanding differences in relative DNA content, chromosome number and morphological features were found. In T. hybridum and partially in T. ambiguum these outstanding differences could be ascribed to variations in accession ploidy level. For the remaining atypical accessions, the determined nuclear DNA content, chromosome number and morphological features were not related to those characteristic of the species. Additionally taxonomic identity of atypical accessions was determined using ITS region sequencing and morphological observations. We propose flow cytometric measurements of nuclear DNA content as simple and reliable technique which can be used at seedling stage to verify identity and genetic stability of Trifolium genebank accessions.     

Dynamics of moisture regime and its reconstruction from a tree-ring width chronology of Pinus sylvestris in the downstream basin of the Selenga River,Russia

Regional tree-ring width chronology of the Scots pine (Pinus sylvestris L.) was constructed from 8 sites in the forest-steppe belt situated in the foothills of the Selenga River basin, Russia. Moisture information contained in tree-ring width chronology was obtained through linear regression reconstruction models of annual August-July precipitation and annual water discharge of the Selenga River during the period 1767-2015. Comparison of the smoothed series allowed estimating long-term variation component of these moisture regime parameters with a high precision. At the same time, regional drought indices are less correlated with pine radial growth, because they have contribution of the other environmental variables, which are much less reflected in the tree-ring of the investigated pine forest stands. Reconstructed dynamic of the moisture regime parameters is supported by documental evident of many socially significant events in the regional history, such as crop failures caused by both droughts and floods, and catastrophic fire in the Irkutsk City in 1879. Also, dependence of the amount of precipitation in the study area on the atmospheric circulation in Central Asia is revealed to have a similar pattern with other regions, i.e., a negative relationship of precipitation with the development of large high atmospheric pressure area within its center in the Altai and Tianshan mountains.     

Oxidative transformation of natural and synthetic phenolic mixtures by Trametes versicolor laccase

The efficiency of Trametes versicolor laccase in the transformation of phenols (caffeic acid, catechol, hydroxytyrosol, methylcatechol, protocatechuic acid, syringic acid, m-tyrosol, 3-hydroxybenzoic acid, 3-hydroxyphenylacetic acid, 2,6-dihydroxybenzoic acid, 4-hydroxybenzaldehyde) usually present in waste water, such as that derived from an olive oil factory, was investigated. According to their response to 24 h laccase action the 11 phenolic compounds were classified in three groups: reactive (88-100% transformation), intermediate reactive (transformation lower than 50%), and recalcitrant (not transformed at all). The enzyme was able to transform the 11 substrates even when they were present in a mixture and also toward a phenolic extract from a Moroccan olive oil mill waste water (OMW) sample. The disappearance of protocatechuic, 3-hydroxyphenylacetic, and 2,6-dihydroxybenzoic acids, and 4-hydroxybenzaldehyde was enhanced whereas that of caffeic acid and m-tyrosol was depressed when the phenols were present in the mixture. A reduction of enzyme activity occurred in single and/or complex phenolic mixtures after enzymatic oxidation. No correspondence between phenol transformation and disappearance of enzymatic activity was, however, observed. The overall results suggest that laccases are effective in the transformation of simple and complex phenolic mixtures.     

Hexavalent Chromium Removal by a Trichoderma inhamatum Fungal Strain Isolated from Tannery Effluent

A fungal strain possibly capable of removing hexavalent chromium was to be isolated from industrial effluent from a leather factory located in the city of Guadalajara, state of Jalisco, Mexico. The strain was identified as Trichoderma inhamatum by the D1/D2 domain sequence of the 28S rDNA gene. Batch cultures of T. inhamatum in media containing initial Cr(VI) concentrations from 0.83 to 2.43 mM Cr(VI) were prepared. Experimental results suggest that the fungus is capable of transforming hexavalent chromium to trivalent chromium; a transformation of a highly toxic contaminant to a low toxic form. The specific and volumetric rates of Cr(VI) reduction by T. inhamatum cultures decreased as the initial Cr(VI) concentration increased. The fungus exhibited a remarkable capacity to tolerate and completely reduce Cr(VI) concentrations up to 2.43 mM. These results indicate that the T. inhamatum fungal strain may have potential applications in bioremediation of Cr(VI)-contaminated wastewaters.     

Transformation of Anaplasma marginale

The tick-borne pathogen, Anaplasma marginale, has a complex life cycle involving ruminants and ixodid ticks. It causes bovine anaplasmosis, a disease with significant economic impact on cattle farming worldwide. The obligate intracellular growth requirement of the bacteria poses a challenging obstacle to their genetic manipulation, a problem shared with other prokaryotes in the genera Anaplasma, Ehrlichia, and Rickettsia. Following our successful transformation of the human anaplasmosis agent, A. phagocytophilum, we produced plasmid constructs (a transposon bearing plasmid, pHimarAm-trTurboGFP-SS, and a transposase expression plasmid, pET28Am-trA7) designed to mediate random insertion of the TurboGFP and spectinomycin/streptomycin resistance genes by the Himar1 allele A7 into the A. marginale chromosome. In these trans constructs, expression of the fluorescent and the selectable markers on the transposon, and expression of the transposase are under control of the A. marginale tr promoter. Constructs were co-electroporated into A. marginale St. Maries purified from tick cell culture, and bacteria incubated for 2 months under selection with a combination of spectinomycin and streptomycin. At that time, ≤1% of tick cells contained colonies of brightly fluorescent Anaplasma, which eventually increased to infect about 80–90% of the cells. Cloning of the insertion site in E. coli and DNA sequence analyses demonstrated insertion of the entire plasmid pHimarAm-trTurboGFP-SS encoding the transposon in frame into the native tr region of A. marginale in an apparent single homologous crossover event not mediated by the transposase. Transformants are fastidious and require longer subculture intervals than wild type A. marginale. This result suggests that A. marginale, as well as possibly other species of Anaplasma and Ehrlichia, can be transformed using a strategy of homologous recombination.