土壤类型对樱桃番茄VC含量及其相关酶活性的影响

以樱桃番茄（Lycopersicon esculentum var. cerasiforme）为材料,研究温室内外及塿土黄绵土条件下,樱桃番茄果实VC含量和抗坏血酸氧化酶（ascorbic acid oxidase,AAO）、抗坏血酸过氧化物酶（ascorbate peroxidase,APX）、多酚氧化酶（polyphenol oxidase,PPO）活性的变化。结果表明：从结果初期到结果末期,VC含量呈先升后降的趋势,AAO活性变化先降后升,APX活性呈上升趋势,PPO活性则随着生育期的进行不断下降。同一环境条件下,塿土樱桃番茄果实VC含量高于黄绵土;同一土类情况下,露天处理优于温室处理。整个生育期内,塿土露天处理樱桃番茄果实VC含量显著高于其他处理,不同处理间差异显著（P＜0.05）。温室黄绵土处理樱桃番茄果实抗坏血酸氧化酶活性最高,露天塿土最低。结果初期APX活性大小顺序为露天塿土＞温室塿土＞温室黄绵土＞露天黄绵土,结果盛期和结果末期均为露天塿土＞露天黄绵土＞温室塿土＞温室黄绵土。而PPO活性在露天黄绵土处理下最高,露天塿土处理次之。果实鲜质量为温室塿土最高。相关分析表明,VC含量与AAO和PPO活性呈负相关。由VC含量与不同酶类之间的关系及各处理对其影响的结果综合分析：在保持相对高VC含量,低AAO、APX和PPO活性的情况下,露天塿土处理最佳。     

Ortho Isomeric Mn(III) N-Alkyl- and Alkoxyalkylpyridylporphyrins—Enhancers of Hyaluronan Degradation Induced by Ascorbate and Cupric Ions

High levels of hyaluronic acid (HA) in tumors correlate with poor outcomes with several types of cancers due to HA-driven support of adhesion, migration and proliferation of cells. In this study we explored how to enhance the degradation of HA into low-molecular fragments, which cannot prevent the immune system to fight tumor proliferation and metastases. The physiological solution of HA was exposed to oxidative degradation by ascorbate and cupric ions in the presence of either one of three ortho isomeric Mn(III) substituted N-alkyl- and alkoxyalkylpyridylporphyrins or para isomeric Mn(III) N-methylpyridyl analog, commonly known as mimics of superoxide dismutase. The changes in hyaluronan degradation kinetics by four Mn(III) porphyrins were monitored by measuring the alteration in the dynamic viscosity of the HA solution. The ortho compounds MnTE-2-PyP⁵⁺ (BMX-010, AEOL10113), MnTnBuOE-2-PyP⁵⁺ (BMX-001) and MnTnHex-2-PyP⁵⁺ are able to redox cycle with ascorbate whereby producing H₂O₂ which is subsequently coupled with Cu(I) to produce the ^•OH radical essential for HA degradation. Conversely, with the para analog, MnTM-4-PyP⁵⁺, no catalysis of HA degradation was demonstrated, due to its inertness towards redox cycling with ascorbate. The impact of different Mn(III)-porphyrins on the HA decay was further clarified by electron paramagnetic resonance spectrometry. The ability to catalyze the degradation of HA in a biological milieu, in the presence of cupric ions and ascorbate under the conditions of high tumor oxidative stress provides further insight into the anticancer potential of redox-active ortho isomeric Mn(III) porphyrins.     

Autoschizis of human ovarian carcinoma cells: scanning electron and light microscopy of a new cell death induced by sodium ascorbate: menadione treatment

Human ovarian carcinoma (MDAH 2774) cells were treated with sodium ascorbate (VC), menadione (VK3), or a combination of both in a ratio 100:1 for 1h and then examined with scanning electron microscopy (SEM) and light microscopy (LM). Light microscopy data corroborated SEM observations, which demonstrated that death of VC+VK3-treated tumor cells occurred primarily by autoschizis. This type of cell death is characterized by a decrease in cell size, cytoplasmic self-excisions, and nuclear and nucleolar morphologic degradations without the formation of apoptotic bodies. Ultimately, cell death results from karyorrhexis and karyolysis. This study illustrates that plasma membrane damage (branching filopodia, blisters, blebs) results from VC treatment; cytoskeletal damage and self-morsellation are caused by VC, VK3 and VC+VK, treatments. The VC treatment results in a 23% decrease in cell diameter while VK3-treated cells decrease cell diameter by 66%. After 1h of VC+VK3 treatment, a heterogenous cell population is found. This population can be resolved into one population whose diameters are 23% smaller than those of sham-treated cells, and a second population whose diameters are approximately twice those of sham-treated cells. This second population is indicative of doublet formation in which the cells appear to be dividing (an early stage of autoschizic cell death). One half of the doublet contains the cell nucleus while the other half consists of cytoplasm and membrane only. The enucleate portion of this doublet will then be excised. When the types of cell death are enumerated following VC+VK3 treatment, 43% of the cells die by autoschizis, 3% by apoptosis, and 1.9% by oncosis. These results confirm that autoschizis is the principal form of cell death that results from the in vitro treatment of human ovarian carcinoma cells with the vitamin combination.     

Lipase-catalyzed synthesis of unsaturated acyl l-ascorbates and their ability to suppress the autoxidation of polyunsaturated fatty acids

l-Ascorbic acid and various polyunsaturated fatty acids (PUFA) were condensed at 55°C by the immobilized lipase Chirazyme l-2 in dry acetone to produce the unsaturated acyl ascorbates. The PUFA moieties of the products were much more resistant to autoxidation at 65°C and nearly 0% relative humidity than the corresponding unmodified PUFA. The effects of the molar ratio of ascorbic acid or linoleoyl ascorbate to linoleic acid on the autoxidation of linoleic acid were examined. The autoxidation of linoleic acid was effectively suppressed at molar ratios greater than or equal to 0.2 when either ascorbic acid or linoleoyl ascorbate was mixed with linoleic acid. The addition of lauroyl ascorbate, synthesized through the enzyme-catalyzed condensation of ascorbic acid and lauric acid in acetone, to docosahexaenoic acid also significantly suppressed the autoxidation of docosahexaenoic acid at molar ratios of ≥0.2.     

Reducing Agent-Mediated Nonenzymatic Conversion of 2-Oxoglutarate to Succinate: Implications for Oxygenase Assays

l -Ascorbate (l -Asc) is often added to assays with isolated Fe^II- and 2-oxoglutarate (2OG)-dependent oxygenases to enhance activity. l -Asc is proposed to be important in catalysis by some 2OG oxygenases in vivo. We report observations on the nonenzymatic conversion of 2OG to succinate, which is mediated by hydrogen peroxide generated by the reaction of l -Asc and dioxygen. Slow nonenzymatic oxidation of 2OG to succinate occurs with some, but not all, other reducing agents commonly used in 2OG oxygenase assays. We intend these observations will help in the robust assignment of substrates and inhibitors for 2OG oxygenases.     

The Epigenetic Role of Vitamin C in Neurodevelopment

The maternal diet during pregnancy is a key determinant of offspring health. Early studies have linked poor maternal nutrition during gestation with a propensity for the development of chronic conditions in offspring. These conditions include cardiovascular disease, type 2 diabetes and even compromised mental health. While multiple factors may contribute to these outcomes, disturbed epigenetic programming during early development is one potential biological mechanism. The epigenome is programmed primarily in utero, and during this time, the developing fetus is highly susceptible to environmental factors such as nutritional insults. During neurodevelopment, epigenetic programming coordinates the formation of primitive central nervous system structures, neurogenesis, and neuroplasticity. Dysregulated epigenetic programming has been implicated in the aetiology of several neurodevelopmental disorders such as Tatton-Brown-Rahman syndrome. Accordingly, there is great interest in determining how maternal nutrient availability in pregnancy might affect the epigenetic status of offspring, and how such influences may present phenotypically. In recent years, a number of epigenetic enzymes that are active during embryonic development have been found to require vitamin C as a cofactor. These enzymes include the ten-eleven translocation methylcytosine dioxygenases (TETs) and the Jumonji C domain-containing histone lysine demethylases that catalyse the oxidative removal of methyl groups on cytosines and histone lysine residues, respectively. These enzymes are integral to epigenetic regulation and have fundamental roles in cellular differentiation, the maintenance of pluripotency and development. The dependence of these enzymes on vitamin C for optimal catalytic activity illustrates a potentially critical contribution of the nutrient during mammalian development. These insights also highlight a potential risk associated with vitamin C insufficiency during pregnancy. The link between vitamin C insufficiency and development is particularly apparent in the context of neurodevelopment and high vitamin C concentrations in the brain are indicative of important functional requirements in this organ. Accordingly, this review considers the evidence for the potential impact of maternal vitamin C status on neurodevelopmental epigenetics.     

Oxidative effects in uninfected tissue in leaves of French bean (Phaseolus vulgaris) containing soft rots caused by Botrytis cinerea

Several markers of oxidative processes have been measured in leaves of Phaseolus vulgaris infected with Botrytis cinerea, with the specific objective of investigating changes induced by this necrotrophic pathogen in tissue remote from the lesion. There was a progressive decrease with time in the contents of ascorbic acid (AA) in apparently healthy tissues from infected plants and non‐inoculated plants grown under identical high‐humidity conditions (abiotically stressed controls), and for periods >48 h this decrease was greater in the infected plants. This decline in AA content was accompanied by an elevation in the intensity of the electron paramagnetic resonance (EPR) signal from adducts of the spin trap α‐(4‐pyridyl‐1‐oxide)‐N‐t‐butylnitrone (POBN), a destabilisation of the (monodehydro) ascorbate radical (Asc^·) signal in the presence of POBN, and an increase in the ratio of Asc^· to AA in samples studied in the absence of the spin trap. These results are consistent with a shift in redox status to more oxidising conditions in apparently healthy tissue of infected plants and indicate the prevalence of chemical processes that are distinctly different from those in uninfected plants. However, no differences in lipid peroxidation products or the single‐peak free radical and Fe(III) (g = 4.27) EPR signals were observed between these tissues distant from the lesions and those from abiotically stressed controls. In addition, the pathogen‐derived sesquiterpene toxin botrydial and a second Mn(II) EPR signal, both of which are associated with Botrytis infection, were not detected in these ‘apparently healthy’ tissues. Copyright © 2003 Society of Chemical Industry     

Characteristic of the Ascorbate Oxidase Gene Family in Beta vulgaris and Analysis of the Role of AAO in Response to Salinity and Drought in Beet

Ascorbate oxidase, which is known to play a key role in regulating the redox state in the apoplast, cell wall metabolism, cell expansion and abiotic stress response in plants, oxidizes apo-plastic ascorbic acid (AA) to dehydroascorbic acid (DHA). However, there is little information about the AAO genes and their functions in beets under abiotic stress. The term salt or drought stress refers to the treatment of plants with slow and gradual salinity/drought. Contrastingly, salt shock consists of exposing plants to high salt levels instantaneously and drought shock occurs under fast drought progression. In the present work, we have subjected plants to salinity or drought treatments to elicit either stress or shock and carried out a genome-wide analysis of ascorbate oxidase (AAO) genes in sugar beet (B. vulgaris cv. Huzar) and its halophytic ancestor (B. maritima). Here, conserved domain analyses showed the existence of twelve BvAAO gene family members in the genome of sugar beet. The BvAAO_1–12 genes are located on chromosomes 4, 5, 6, 8 and 9. The phylogenetic tree exhibited the close relationships between BvAAO_1-12 and AAO genes of Spinacia oleracea and Chenopodium quinoa. In both beet genotypes, downregulation of AAO gene expression with the duration of salt stress or drought treatment was observed. This correlated with a decrease in AAO enzyme activity under defined experimental setup. Under salinity, the key downregulated gene was BvAAO_10 in Beta maritima and under drought the BvAAO_3 gene in both beets. This phenomenon may be involved in determining the high tolerance of beet to salinity and drought.     

Friend or Foe: The Relativity of (Anti)oxidative Agents and Pathways

An element, iron, a process, the generation of reactive oxygen species (ROS), and a molecule, ascorbate, were chosen in our study to show their dual functions and their role in cell fate decision. Iron is a critical component of numerous proteins involved in metabolism and detoxification. On the other hand, excessive amounts of free iron in the presence of oxygen can promote the production of potentially toxic ROS. They can result in persistent oxidative stress, which in turn can lead to damage and cell death. At the same time, ROS—at strictly regulated levels—are essential to maintaining the redox homeostasis, and they are engaged in many cellular signaling pathways, so their total elimination is not expedient. Ascorbate establishes a special link between ROS generation/elimination and cell death. At low concentrations, it behaves as an excellent antioxidant and has an important role in ROS elimination. However, at high concentrations, in the presence of transition metals such as iron, it drives the generation of ROS. In the term of the dual function of these molecules and oxidative stress, ascorbate/ROS-driven cell deaths are not necessarily harmful processes—they can be live-savers too.