HPLC Analysis of Phenolic Compounds and Antioxidant Activity of Ribes Fruit Leaves

Berry fruit leaves are recognized as potential medicaments which are rich in different phenolic compounds, and have been used in folk medicine for centuries. In order to evaluate phenol composition of fruit leaves from red currant (Ribes rubrum) and black currant (Ribes nigrum) species were subjected to the spectrophotometric and HPLC analysis. The antioxidant activity was estimated using DPPH free radical scavenging assay. Flavonols, flavan-3-ols and phenolic acids were the main phenol classes found in the investigated leaf extracts. Investigated extracts showed significant antioxidant activity and a correlation with total phenol content. Ribes leaf extracts, rich in phenolic content, with significant antioxidant activity, can be used as medical supplements

Polyphenols and Antioxidant Activity of Different Vinegrape Leaves

The volume of the grape waste produced directly relates to the volume of grapes pressed and in turn depends on the specific climatic conditions of the relevant vintage. The aim of this work is to evaluate vinegrape leaves as a potential source of natural antioxidants – polyphenols for their possible use as dietary supplement or food antioxidants. To this purpose antioxidant activity and contents of phenolic compounds of the leaves extracts of five grape varieties of Vitis vinifera L (Vranac, Prokupac, Merlot, Gamay and Italian Rizling), grown in southern Serbia were investigated. The analysis show high content of polyphenols reflecting their high antioxidant activity (R2 = 0.9819, p<0.01). According to the obtained results, the leaves extracts, can be considered rich natural source of phenolic compounds with good antioxidant properties

Developmental arrest in Drosophila melanogaster caused by mitochondrial DNA replication defects cannot be rescued by the alternative oxidase

The xenotopic expression of the alternative oxidase AOX from the tunicate Ciona intestinalis in diverse models of human disease partially alleviates the phenotypic effects of mitochondrial respiratory chain defects. AOX is a non-proton pumping, mitochondrial inner membrane-bound, single-subunit enzyme that can bypass electron transport through the cytochrome segment, providing an additional site for ubiquinone reoxidation and oxygen reduction upon respiratory chain overload. We set out to investigate whether AOX expression in Drosophila could counteract the effects of mitochondrial DNA (mtDNA) replication defects caused by disturbances in the mtDNA helicase or DNA polymerase gamma. We observed that the developmental arrest imposed by either the expression of mutant forms of these enzymes or their knockdown was not rescued by AOX. Considering also the inability of AOX to ameliorate the phenotype of tko(25t), a fly mutant with mitochondrial translation deficiency, we infer that this alternative enzyme may not be applicable to cases of mitochondrial gene expression defects. Finding the limitations of AOX applicability will help establish the parameters for the future putative use of this enzyme in gene therapies for human mitochondrial diseases.Peer reviewe

Expression of Ciona intestinalis AOX causes male reproductive defects in Drosophila melanogaster

Background: Mitochondrial alternative respiratory-chain enzymes are phylogenetically widespread, and buffer stresses affecting oxidative phosphorylation in species that possess them. However, they have been lost in the evolutionary lineages leading to vertebrates and arthropods, raising the question as to what survival or reproductive disadvantages they confer. Recent interest in using them in therapy lends a biomedical dimension to this question. Methods: Here, we examined the impact of the expression of Ciona intestinalis alternative oxidase, AOX, on the reproductive success of Drosophila melanogaster males. Sperm-competition assays were performed between flies carrying three copies of a ubiquitously expressed AOX construct, driven by the a-tubulin promoter, and wild-type males of the same genetic background. Results: In sperm-competition assays, AOX conferred a substantial disadvantage, associated with decreased production of mature sperm. Sperm differentiation appeared to proceed until the last stages, but was spatially deranged, with spermatozoids retained in the testis instead of being released to the seminal vesicle. High AOX expression was detected in the outermost cell-layer of the testis sheath, which we hypothesize may disrupt a signal required for sperm maturation. Conclusions: AOX expression in Drosophila thus has effects that are deleterious to male reproductive function. Our results imply that AOX therapy must be developed with caution.Peer reviewe

Expression of Ciona intestinalis AOX causes male reproductive defects in Drosophila melanogaster

Background: Mitochondrial alternative respiratory-chain enzymes are phylogenetically widespread, and buffer stresses affecting oxidative phosphorylation in species that possess them. However, they have been lost in the evolutionary lineages leading to vertebrates and arthropods, raising the question as to what survival or reproductive disadvantages they confer. Recent interest in using them in therapy lends a biomedical dimension to this question. Methods: Here, we examined the impact of the expression of Ciona intestinalis alternative oxidase, AOX, on the reproductive success of Drosophila melanogaster males. Sperm-competition assays were performed between flies carrying three copies of a ubiquitously expressed AOX construct, driven by the a-tubulin promoter, and wild-type males of the same genetic background. Results: In sperm-competition assays, AOX conferred a substantial disadvantage, associated with decreased production of mature sperm. Sperm differentiation appeared to proceed until the last stages, but was spatially deranged, with spermatozoids retained in the testis instead of being released to the seminal vesicle. High AOX expression was detected in the outermost cell-layer of the testis sheath, which we hypothesize may disrupt a signal required for sperm maturation. Conclusions: AOX expression in Drosophila thus has effects that are deleterious to male reproductive function. Our results imply that AOX therapy must be developed with caution.Peer reviewe

Ligand-Bound GeneSwitch Causes Developmental Aberrations in Drosophila that Are Alleviated by the Alternative Oxidase

Culture of Drosophila expressing the steroid-dependent GeneSwitch transcriptional activator under the control of the ubiquitous -tubulin promoter was found to produce extensive pupal lethality, as well as a range of dysmorphic adult phenotypes, in the presence of high concentrations of the inducing drug RU486. Prominent among these was cleft thorax, seen previously in flies bearing mutant alleles of the nuclear receptor Ultraspiracle and many other mutants, as well as notched wings, leg malformations, and bristle abnormalities. Neither the -tubulin-GeneSwitch driver nor the inducing drug on their own produced any of these effects. A second GeneSwitch driver, under the control of the daughterless promoter, which gave much lower and more tissue-restricted transgene expression, exhibited only mild bristle abnormalities in the presence of high levels of RU486. Coexpression of the alternative oxidase (AOX) from Ciona intestinalis produced a substantial shift in the developmental outcome toward a wild-type phenotype, which was dependent on the AOX expression level. Neither an enzymatically inactivated variant of AOX, nor GFP, or the alternative NADH dehydrogenase Ndi1 from yeast gave any such rescue. Users of the GeneSwitch system should be aware of the potential confounding effects of its application in developmental studies.Peer reviewe

Effects on Dopaminergic Neurons Are Secondary in COX-Deficient Locomotor Dysfunction in Drosophila

Dopaminergic (DA) neurons have been implicated as key targets in neurological disorders, notably those involving locomotor impairment, and are considered to be highly vulnerable to mitochondrial dysfunction, a common feature of such diseases. Here we investigated a Drosophila model of locomotor disorders in which functional impairment is brought about by pan-neuronal RNAi knockdown of subunit COX7A of cytochrome oxidase (COX). Despite minimal neuronal loss by apoptosis, the expression and activity of tyrosine hydroxylase was decreased by half. Surprisingly, COX7A knockdown specifically targeted to DA neurons did not produce locomotor defect. Instead, using various drivers, we found that COX7A knockdown in specific groups of cholinergic and glutamatergic neurons underlay the phenotype. Based on our main finding, the vulnerability of DA neurons to mitochondrial dysfunction as a cause of impaired locomotion in other organisms, including mammals, warrants detailed investigation.Peer reviewe

Diiron centre mutations in Ciona intestinalis alternative oxidase abolish enzymatic activity and prevent rescue of cytochrome oxidase deficiency in flies

The mitochondrial alternative oxidase, AOX, carries out the non proton-motive re-oxidation of ubiquinol by oxygen in lower eukaryotes, plants and some animals. Here we created a modified version of AOX from Ciona instestinalis, carrying mutations at conserved residues predicted to be required for chelation of the diiron prosthetic group. The modified protein was stably expressed in mammalian cells or flies, but lacked enzymatic activity and was unable to rescue the phenotypes of flies knocked down for a subunit of cytochrome oxidase. The mutated AOX transgene is thus a potentially useful tool in studies of the physiological effects of AOX expression.Peer reviewe

Genetic and phenotypic evaluation of european maize landraces as a tool for conservation and valorization of agrobiodiversity

The ECPGR European Evaluation Network (EVA) for Maize involves genebanks, research institutions, and private breeding companies from nine countries focusing on the valorization of maize genetic resources across Europe. This study describes a diverse collection of 626 local landraces and traditional varieties of maize (Zea mays L.) from nine European genebanks, including criteria for selection of the collection and its genetic and phenotypic diversity. High-throughput pool genotyping grouped the landraces into nine genetic groups with a threshold of 0.6 admixture, while 277 accessions were designated admixed and likely to have resulted from previous breeding activities. The grouping correlated well with the geographic origins of the collection, also reflecting the various pathways of introduction of maize to Europe. Phenotypic evaluations of 588 accessions for flowering time and plant architecture in multilocation trials over three years confirmed the great diversity within the collection, although phenotypic clusters only partially correlated with the genetic grouping. The EVA approach promotes conservation of genetic resources and opens an opportunity to increase genetic variability for developing improved varieties and populations for farmers, with better adaptation to specific environments and greater tolerance to various stresses. As such, the EVA maize collection provides valuable sources of diversity for facing climate change due to the varieties’ local adaptation

Hlorofil kao merilo zdravlja biljaka: agroekološki aspekti

As photosynthesis is the basic process during which light energy is absorbed and converted into organic matter, the importance of the plant pigment chlorophyll (a and b forms) as an intermediary in transformation of the absorbed solar energy and its activity in the process of photosynthesis and synthesis of organic substances in plants are crucial. Therefore, this paper provides an overview of methods for monitoring the optical activity of chlorophyll molecules and methods (non-destructive and destructive) for quantification of chlorophyll in plants. These methods are used to estimate the effects of different stress factors (abiotic, biotic and xenobiotic) on the efficiency of photosynthesis and bioproductivity, aiming to assess the impact that these limiting factors have on the yield of various cultivars. Also, those methods for analysis of chlorophyll optical activity and/or content are appropriate for assessing the reaction of weed species to different agricultural practices (mineral nutrition, treatment by herbicides, etc.) and studies of different aspects of weed ecophysiology and their influence on crop harvest.Kako je proces fotosinteze osnovni proces tokom kojeg se svetlosna energija apsorbuje i konvertuje u organsku materiju, ključni je značaj postojanja biljnog pigmenta hlorofila (a i b forma) kao posrednika u transformaciji apsorbovane svetlosne energije i njegove aktivnosti u procesu fotosinteze i sinteze organskih materija kod biljaka. Stoga je u radu dat pregled metoda za praćenje optičke aktivnosti molekula hlorofila, kao i metoda (nedestruktivnih i destruktivnih) kvantifikacije hlorofila u biljkama. Ove metode se primenjuju u proceni uticaja različitih stresnih faktora (abiotskih, biotskih i ksenobiotskih) na efikasnost fotosinteze i bioproduktivnost biljaka, sa ciljem procenjivanja uticaja koji ovi ograničavajući faktori imaju na prinos useva. Takođe, pomenute metode za analizu optičke aktivnosti i/ili sadržaja hlorofila su odgovarajuće i za procenu reakcije korova na različite poljoprivredne prakse (mineralna ishrana, primena herbicida i sl.) i ispitivanje različitih aspekata ekofiziologije korova i procenu njihovog uticaja na prinos useva