Higher Reactive Oxygen Species Content Research Articles

Multifunctional visible light photocatalytic carbon dots synergize with reactive oxygen species for anti-quorum sensing and anti-bacteria for salmon preservation

Quorum sensing (QS)-induced spoilage causes approximately 15–20% of annual food wastage. However, traditional natural QS inhibitors exhibit poor water solubility and no catalytic activity. Therefore, this study aimed to synthesize novel anti-QS carbon dots (RR-CDs) with good water solubility, visible light photocatalytic activity, and high biocompatibility. The RR-CDs (<200 μg mL−1) exhibited considerable broad-spectrum antibacterial activity by generating reactive oxygen species (ROS) to disrupt the cell membrane wall structure. Owing to the appropriate bandgap of RR-CDs, a high ROS content, including ·O2−, 1O2, and ·OH, was generated via energy transfer under visible light irradiation. Furthermore, the combined effect of RR-CDs and ROS reduced the biofilm formation, swarming, swimming, and extracellular protease production of Pseudomonas fluorescens by 48.45%, 25.51%, 52.79%, and 38.67%, respectively, by inhibiting LuxI/R expression, which further decreased pgaC, tyrB, motB, and aprE expression by >50 %. Notably, RR-CDs exhibited no obvious cytotoxicity; furthermore, they could extend the shelf life of salmon by 3 d. Altogether, these results present and validate the strong ability of photocatalytic RR-CDs for QS inhibition and show their potential in aquatic product preservation.

Vascular Regenerative Cell Deficiencies in South Asian Adults

BackgroundSouth Asian individuals shoulder a disproportionate burden of cardiometabolic diseases. ObjectivesThe purpose of this study was to determine if vascular regenerative cell content varies significantly between South Asian and White European people. MethodsBetween January 2022 and January 2023, 60 South Asian and 60 White European adults with either documented cardiovascular disease or established diabetes with ≥1 other cardiovascular risk factor were prospectively enrolled. Vascular regenerative cell content in venous blood was enumerated using a flow cytometry assay that is based on high aldehyde dehydrogenase (ALDHhi) activity and cell surface marker phenotyping. The primary outcome was the difference in frequency of circulating ALDHhi progenitor cells, monocytes, and granulocytes between the 2 groups. ResultsCompared with White European participants, those of South Asian ethnicity were younger (69 ± 10 years vs 66 ± 9 years; P < 0.05), had lower weight (88 ± 19 kg vs 75 ± 13 kg; P < 0.001), and exhibited a greater prevalence of type 2 diabetes (62% vs 92%). South Asian individuals had markedly lower circulating frequencies of pro-angiogenic ALDHhiSSClowCD133+ progenitor cells (P < 0.001) and ALDHhiSSCmidCD14+CD163+ monocytes with vessel-reparative capacity (P < 0.001), as well as proportionally more ALDHhi progenitor cells with high reactive oxygen species content (P < 0.05). After correction for sex, age, body mass index, and glycated hemoglobin, South Asian ethnicity was independently associated with lower ALDHhiSSClowCD133+ cell count. ConclusionsSouth Asian people with cardiometabolic disease had less vascular regenerative and reparative cells suggesting compromised vessel repair capabilities that may contribute to the excess vascular risk in this population. (The Role of South Asian vs European Origins on Circulating Regenerative Cell Exhaustion [ORIGINS-RCE]; NCT05253521)

Uncovering the mechanisms underlying pear leaf apoplast protein-mediated resistance against Colletotrichum fructicola through transcriptome and proteome profiling

Pear anthracnose, caused by the fungus Colletotrichum fructicola, is a devastating disease for the pear industry. The apoplast, an extracellular compartment outside the plasma membrane, plays a crucial role in water and nutrient transport, as well as plant-microbe interactions. This study aimed to uncover the molecular mechanism of pear leaf apoplastic protein-mediated resistance to C. fructicola. Apoplast fluid was isolated using the vacuum infiltration method, and defence-related apoplastic proteins were identified through protein mass spectrometry and transcriptome sequencing. We found 213 apoplastic proteins in the leaf apoplast fluid during early C. fructicola infection, with the majority (74.64%) being enzymes, including glycosidases, proteases, and oxidoreductases. Gene Ontology analysis revealed their involvement in defence response, enzyme inhibition, carbohydrate metabolism, and phenylpropanoid biosynthesis. Transcriptome analysis showed the infection induced expression of certain apoplast proteins, potentially contributing to pear leaf resistance. Notably, the expression of PbrGlu1, an endo-β-1,3-glucanase from the glycoside hydrolase 17 family, was significantly higher in infected leaves. Silencing of the PbrGlu1 gene increased pear leaf susceptibility to C. fructicola, leading to more severe symptoms and higher reactive oxygen species content. Overall, our study provides insights into the apoplast space interaction between pear leaves and C. fructicola, identifies a key gene in infected pears, and offers a foundation and new strategy for understanding the molecular mechanisms underlying pear anthracnose and breeding disease-resistant pears.

Multi-omics insights into the positive role of strigolactone perception in barley drought response

BackgroundDrought is a major environmental stress that affects crop productivity worldwide. Although previous research demonstrated links between strigolactones (SLs) and drought, here we used barley (Hordeum vulgare) SL-insensitive mutant hvd14 (dwarf14) to scrutinize the SL-dependent mechanisms associated with water deficit response.ResultsWe have employed a combination of transcriptomics, proteomics, phytohormonomics analyses, and physiological data to unravel differences between wild-type and hvd14 plants under drought. Our research revealed that drought sensitivity of hvd14 is related to weaker induction of abscisic acid-responsive genes/proteins, lower jasmonic acid content, higher reactive oxygen species content, and lower wax biosynthetic and deposition mechanisms than wild-type plants. In addition, we identified a set of transcription factors (TFs) that are exclusively drought-induced in the wild-type barley.ConclusionsCritically, we resolved a comprehensive series of interactions between the drought-induced barley transcriptome and proteome responses, allowing us to understand the profound effects of SLs in alleviating water-limiting conditions. Several new avenues have opened for developing barley more resilient to drought through the information provided. Moreover, our study contributes to a better understanding of the complex interplay between genes, proteins, and hormones in response to drought, and underscores the importance of a multidisciplinary approach to studying plant stress response mechanisms.

Mechanism of time-dependent toxicity of quinolone antibiotics on luminescent bacteria Vibrio qinghaiensis sp.-Q67

Four quinolone antibiotics (ciprofloxacin (CIP), enrofloxacin (ENR), sparfloxacin (SPA), gatifloxacin (GAT)) and their binary mixtures at environmentally relevant concentrations exhibited time-dependent hormesis on Vibrio qinghaiensis sp.-Q67 (Q67). The study aims to investigate the time-dependent toxicity of low-dose pollutants and the occurrence of hormesis. These indicators, total protein (TP), reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and luminescence-related chemicals flavin mononucleotide (FMN), nicotinamide adenine dinucleotide (NADH), were measured to explore the mechanism of hormesis. The results showed a trend of increases in all indicators after 12 h of exposure, reaching maximal effects at 60 h and then decreasing as time progressed. At 36 h, 60 h and 84 h, the results showed a gradual increase followed by a decreasing trend in TP, FMN and NADH as the concentration in the group increased, whereas ROS, CAT, SOD and MDA showed the opposite trend. Notably, the degree of changes was related to the magnitude of hormesis. At low concentrations, the content of ROS and MDA decreased, the activity of CAT and SOD was lower, but the content of TP, FMN, NADH gradually increased, positively correlated with the promotion of Q67. At high concentrations, ROS and MDA content in Q67 increased, triggering the antioxidant defense mechanism (CAT and SOD activity increased), but TP, FMN, NADH content decreased, negatively correlated with the inhibited Q67. Therefore, our findings demonstrated two common patterns in these seven biochemical indicators on Q67. These findings have important practical implications for the ecological risk assessment of antibiotics in aquatic environment.

Acute toxicity, oxidative stress, and apoptosis due to short-term triclosan exposure and multi- and transgenerational effects on in vivo endpoints, antioxidant defense, and DNA damage response in the freshwater water flea Daphnia magna

In this study, we measured the acute toxicity of triclosan (TCS) in neonate and adult Daphnia magna water fleas. The median lethal concentrations were 184.689 and 349.511 μg/L, respectively. Oxidative stress induced by TCS was analyzed based on changes in reactive oxygen species (ROS) content and antioxidant enzymatic activities in D. magna. Based on these endpoints, TCS concentrations of 50 and 100 μg/L induced oxidative stress. However, several apoptosis-mediated proteins showed TCS-induced oxidative-stress damage in response to 25 μg/L, indicating that apoptotic proteins were the most sensitive mediators. We also evaluated the multi- and transgenerational effects of TCS on D. magna over three generations in terms of various in vivo endpoints, DNA damage responses, and biochemical reactions. The transgenerational group exposed to TCS exhibited greater negative impacts on antioxidant responses, DNA fragmentation status, and biological endpoints compared with the multigenerational exposure group, leading to decreased reproductive rates and higher ROS content. The transcriptional expression levels of glutathione S-transferase genes in the transgenerational exposure group were upregulated compared to those in the multigenerational group but were fully recovered in F2 offspring. Our findings provide an in-depth understanding of the adaptive effects of multigenerational exposure to TCS.

Metagenomic analysis and nitrogen removal performance evaluation of activated sludge from a sequencing batch reactor under different salinities

The effect of salinity on the nitrogen removal performance and microbial community of activated sludge was investigated in a sequencing batch reactor. The NH4+-N removal efficiency was over 95% at 0–4% salinity, indicating that the nitrification performance of activated sludge was slightly affected by lower salinity. The obvious nitrite accumulation was observed with the increment of the salinity to 5%, followed by a notable decline in the nitrogen removal performance at 6% salinity. The salinity inhibited the microbial activity, and the specific rate of nitrification and denitrification was decreased by the increasing salinity obviously. Additionally, the lower activity of superoxide dismutase and peroxidase and higher reactive oxygen species content in activated sludge might account for the deteriorative nitrogen removal performance at 6% salinity. Metagenomics analysis revealed that the genes encoding the ABC-type quaternary amine transporter in the ABC transporter pathway were abundant in the activated sludge at 2% and 4% salinity, and the higher salinity of 6% led to the loss of the genes encoding the p-type Na+ transporter in the ABC transporter pathway. These results indicated that the salinity could weaken the ABC transporter pathway for the balance of osmotic pressure in activated sludge. The microbial activity and nitrogen removal performance of activated sludge were decreased due to the unbalanced osmotic pressure at higher salinity.

The Impact of the Animal Housing System on Immune Cell Composition and Function in the Blood of Dromedary Camels

Simple SummaryThe present study investigated the impacts of a change in animal housing system on selected parameters of the camel immune system. Samples collected from camels during a free-ranging time were compared with samples collected from the same camels during movement-restricted housing. Movement-restricted camels showed elevated myeloperoxidase activity in their serum, a significant shape-change of their neutrophils, and higher reactive oxygen species content in their monocytes and neutrophils. The leukogram pattern of the camels under restricted housing was characterized by increased numbers of neutrophils, eosinophils, lymphocytes, and monocytes. Within the lymphocyte population, only the helper T cells and B cells were expanded in animals under restricted housing. In addition, restricted housing modulated the expression of several cell surface antigens, including monocyte-polarization markers and cell adhesion molecules. Functional analysis of bacterial phagocytosis indicated impaired antibacterial function of phagocytes in camels under restricted housing. In summary, the present study identified significant changes in blood immune cell composition, phenotype, and function in dromedary camels under restricted-housing conditions, and suggests the development of an excitement leukogram in those animals.Background: The dromedary camel (Camelus dromedarius) is an important livestock animal of desert and semi-desert ecosystems. In recent years, several elements of the camel immune system have been characterized. Stress and excitement induced by animal housing represent the most important environmental factors with potential modulatory effects on the immune system. The present study evaluated the impacts of a restricted-housing system on some phenotypic and functional properties of blood leukocytes in dromedary camels. Methods: Immunofluorescence and flow cytometry were used to comparatively analyze samples collected from camels during a free-ranging time and samples collected from the same camels during movement-restricted housing. Results: In comparison to blood samples collected from the camels during the free-ranging time, samples from movement-restricted camels showed elevated serum myeloperoxidase activity, a significant shape-change in their neutrophils, and higher reactive oxygen species content in their monocytes and neutrophils, indicating increased cellular oxidative stress under movement-restricted housing. The leukogram pattern of the camels under restricted housing was characterized by leukocytosis with increased numbers of neutrophils, eosinophils, lymphocytes, and monocytes, resembling an excitement leukogram pattern. Within the lymphocyte population, only the helper T cells and B cells were expanded in animals under restricted housing. The upregulation of CD163 together with the downregulation of MHC-II on monocytes from excited camels indicate a modulatory potential of animal excitement to polarize monocytes toward an anti-inflammatory phenotype. Functional analysis of bacterial phagocytosis indicates an impaired antibacterial function of phagocytes in excited camels. The downregulation of several cell adhesion molecules on leukocytes from excited camels suggests a role for impaired cell adhesion and tissue migration and leukocyte retention in blood in the observed leukocytosis in animals under excitement. Conclusions: The present study identified significant changes in blood immune cell composition, phenotype, and function in dromedary camels under restricted-housing conditions. The observed changes in leukocyte composition suggest the development of an excitement leukogram pattern in camels under movement-restricted housing. To evaluate the clinical relevance of the observed changes in immune cell phenotype and function for the immune competence of camels under restricted housing, further studies are required.

Cannabidiol converts NF-κB into a tumor suppressor in glioblastoma with defined antioxidative properties.

BackgroundThe transcription factor NF-κB drives neoplastic progression of many cancers including primary brain tumors (glioblastoma [GBM]). Precise therapeutic modulation of NF-κB activity can suppress central oncogenic signaling pathways in GBM, but clinically applicable compounds to achieve this goal have remained elusive.MethodsIn a pharmacogenomics study with a panel of transgenic glioma cells, we observed that NF-κB can be converted into a tumor suppressor by the non-psychotropic cannabinoid cannabidiol (CBD). Subsequently, we investigated the anti-tumor effects of CBD, which is used as an anticonvulsive drug (Epidiolex) in pediatric neurology, in a larger set of human primary GBM stem-like cells (hGSC). For this study, we performed pharmacological assays, gene expression profiling, biochemical, and cell-biological experiments. We validated our findings using orthotopic in vivo models and bioinformatics analysis of human GBM datasets.ResultsWe found that CBD promotes DNA binding of the NF-κB subunit RELA and simultaneously prevents RELA phosphorylation on serine-311, a key residue that permits genetic transactivation. Strikingly, sustained DNA binding by RELA-lacking phospho-serine 311 was found to mediate hGSC cytotoxicity. Widespread sensitivity to CBD was observed in a cohort of hGSC defined by low levels of reactive oxygen species (ROS), while high ROS content in other tumors blocked CBD-induced hGSC death. Consequently, ROS levels served as a predictive biomarker for CBD-sensitive tumors.ConclusionsThis evidence demonstrates how a clinically approved drug can convert NF-κB into a tumor suppressor and suggests a promising repurposing option for GBM therapy.

Light modulates the effect of antibiotic norfloxacin on photosynthetic processes of Microcystis aeruginosa.

Norfloxacin is one of the widely used antibiotics, often detected in aquatic ecosystems, and difficultly degraded in the environment. However, how norfloxacin affects the photosynthetic process of freshwater phytoplankton is still largely unknown, especially under varied light conditions. In this study, we investigated photosynthetic mechanisms of Microcystis aeruginosa in responses to antibiotic norfloxacin (0-50 μg/L) for 72 h under low (LL; 50 μmol photons m-2 s-1) and high (HL; 250 μmol photons m-2 s-1) growth light regimes. We found that environmentally related concentrations of norfloxacin inhibited the growth rate and operational quantum yield of photosynthesis system II (PSII) of M. aeruginosa more under HL than under LL, suggesting HL increased the toxicity of norfloxacin to M. aeruginosa. Further analyses showed that norfloxacin deactivated PSII reaction centers under both growth light regimes with increased minimal fluorescence yields only under HL, suggesting that norfloxacin not only damaged reaction centers of PSII, but also inhibited energy transfer among phycobilisomes in M. aeruginosa under HL. However, non-photosynthetic quenching decreased in the studied species by norfloxacin exposure under both growth light regimes, suggesting that excess energy might not be efficiently dissipated as heat. Also, we found that reactive oxygen species (ROS) content increased under norfloxacin treatments with a higher ROS content under HL compared to LL. In addition, HL increased the absorption of norfloxacin by M. aeruginosa, which could partly explain the high sensitivity to norfloxacin of M. aeruginosa under HL. This study firstly reports that light can strongly affect the toxicity of norfloxacin to M. aeruginosa, and has vitally important implications for assessing the toxicity of norfloxacin to aquatic microorganisms.

Lycopene Alleviates Hepatic Hypoxia/Reoxygenation Injury Through Nrf2/HO-1 Pathway in AML12 Cell.

[Figure: see text] Lycopene (lyc) has an effect on preventing cancer, yet its effects on hypoxia/reoxygenation (H/R) injury remained obscure. The study aimed at discovering its role in preventing hepatic cells against H/R injury. Hepatic cells were incubated in hypoxia incubator to simulate ischemia/reperfusion injury in vitro. Cell viability was detected using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after Lycopene treatment with or without ML385 (nuclear factor erythroid 2-related factor 2 [Nrf2] inhibitor). Lactate dehydrogenase (LDH) and malondialdehyde (MDA) content were detected. Cellular cytokine (tumor necrosis factor-α, TNF-α; interleukin-6, IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA). Hepatic cell apoptosis and cellular reactive oxygen species (ROS) content was detected by flow cytometry. Nrf2 transfer was observed using immunofluorescence staining. Nrf2 and heme oxygenase-1 (HO-1) expressions were detected with quantitative real-time polymerase chain reaction and western blot as needed. In hepatic cells, after H/R, the viability was dropped, TNF-α and IL-6 levels and LDH and MDA content were increased, with high apoptosis rate and ROS content. Lycopene led to a reversed effect, with promotion on Nrf2 transfer from cytoplasm into nucleus and Nrf2/HO-1 pathway activation. Further experiments showed that ML385 could reverse the effects of Lycopene. Lycopene could activate Nrf2/HO-1 pathway to protect hepatic cells against H/R injury.

Chemical composition and cytotoxic screening of Musa cavendish green peels extract: Antiproliferative activity by activation of different cellular death types

Musa cavendish, commonly known as banana, is a fruit with nutritional and therapeutic properties. We investigated the chemical composition and in vitro cytotoxic effect of M. cavendish green peel extract (MHE) on cancer cells for the first time. The compounds characterization was performed by HPLC-UV/Vis and FIA-ESI-IT-MSn. We investigated in vitro cytotoxic effect of Musa cavendish green peels extract (MHE) in HepG2, A-375, MCF-7 and Caco-2 cancer cells. We evaluated the effect of MHE on proliferation of different cell lines through apoptosis, necrosis, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) content determination. We identified 12 compounds from different classes in the extract, including derivatives of phenolic acids, aglycone flavonoids, glycoside flavonoids and catecholamines. Our results indicate that MHE exerts, after 48 h treatment, an accentuated antiproliferative effect from the dose of 100 μg/mL in all cell lines tested. In HepG2 cells, these effects were related to the induction of cell death, both necrotic and apoptotic, and remarkable changes in cell morphology. Depolarization of MMP and high ROS content were also observed in the cells in a dose-dependent manner. Our results show that MHE may be used as a source of new drugs with anticancer activity.

Effects of Various Light-Emitting Diode (LEd) Wavelengths on the Growth of Scenedesmus Obliquus Fachb-12 and Accumulation of Astaxanthin

Given the central role of light in the algal photosynthesis, respiration, cell division, growth and the accumulation of value products, the effects of light-emitting diodes (LEDs) light wavelengths (blue, white, red and green) were studied in Scenedesmus obliquus. Biomass, residual nutrient amount, soluble protein, astaxanthin and reactive oxygen species, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activity were analyzed to determine the effects of different monochromatic light wavelengths via biochemical methods. The results showed that blue light wavelength is the optimal light wavelength for phosphorus removal efficiency and the accumulation of biomass and astaxanthin in S. obliquus. Meanwhile, high reactive oxygen species content under the blue light might induce the accumulation of astaxanthin. The high activity of SOD, CAT and POD might participate in clearing the reactive oxygen species to facilitate the growth of microalgae. Furthermore, we found mixed blue/green lights treatment is the most appropriate mixture for the nitrogen removal. Under the blue light treatment, high light intensity and 18L:6D light cycle is the best condition for biomass and astaxanthin accumulation. Optimal nitrogen/phosphorus removal efficiency was observed under a 24L:0D light cycle. These results might provide a foundational data for the optimizing the productivity of high-value metabolites and treatment of wastewater.

An Endogenous Reactive Oxygen Species (ROS)‐Activated Histone Deacetylase Inhibitor Prodrug for Cancer Chemotherapy

Suberoylanilide hydroxamic acid (SAHA, vorinostat) is a potent small-molecule pan-inhibitor of histone deacetylases (HDACs) approved for treatment of cutaneous T-cell lymphoma (CTCL). However, SAHA exhibits poor selectivity for cancer cells over noncancer cells. With an aim to improving its selectivity for cancer cells, we generated a novel SAHA prodrug (SAHA-OBP) that is activated in the presence of hydrogen peroxide, a reactive oxygen species (ROS) known to be overexpressed in cancer cells. The high endogenous ROS content in cancer cells triggers rapid removal of the 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl carbonyl (OBP) cap to release active SAHA. The SAHA-OBP prodrug demonstrates selective activity against multiple cancer cell lines such as HeLa, MCF-7, MDA-MB-231, and B16-F10, while remaining benign toward noncancer cells. The downstream effects of SAHA released from SAHA-OBP in cancer cells is the induction of apoptosis. SAHA-OBP was also found to be effective on multicellular tumor spheroids (MCTS). The SAHA prodrug designed in this study undergoes rapid ROS-dependent activation and imparts much-needed selectivity to SAHA for cancer cells.

Role of reactive oxygen species in the anticancer activity of botanicals: Comparing sensitivity profiles.

Numerous botanicals have been shown to exhibit in vitro and in vivo anticancer activity, some of which is the result of the induction of reactive oxygen species (ROS) in cancer cells with a high ROS content. The present study compared sensitivities to a series of botanicals among cancer cell lines, using an XTT viability test, in order to create a specific cancer-herb profile. Of the 27 botanicals screened, 10 exhibited a cytotoxic effect, 7 of which were ROS-mediated. The sensitivity profiles of the ROS-inducing botanicals in 10 cancer cell lines were similar, unlike 3 cytotoxic ROS-independent botanicals that displayed divergent botanical-specific profiles. The correlation between sensitivity profiles of ROS-inducing botanicals suggests a common mechanism of action, in contrast to the varied mechanism of ROS-independent botanicals. This implies that the investigation of the anticancer activity of botanicals should start with the examination of ROS-mediated activity. Further investigation of ROS sensitivity among various tumor types is required in order to guide research into developing evidence-based guidelines in the use of botanicals for cancer treatment.

Development of resistance to antiglioma agents in rat C6 cells caused collateral sensitivity to doxorubicin

Chemoresistance is a severe limitation to glioblastoma (GBM) therapy and there is a strong need to understand the underlying mechanisms that determine its response to different chemotherapeutics. Therefore, we induced resistance in C6 rat glioma cell line, which considerably resembles the characteristics of human GBM. The resistant phenotype was developed by 3-bis (2-chloroethyl)-1-nitrosourea (BCNU), one of the most commonly used therapeutic drug in the course of GBM treatment. After confirmation of the cross-resistance to cisplatin (CPt) and temozolomide (TMZ) in newly established RC6 cell line, we examined cell death induction and DNA damage by these drugs. Resistance to apoptosis and deficiency in forming DNA double-strand breaks was followed by significant decrease in the mRNA expression of pro-apoptotic and anti-apoptotic genes. The development of drug resistance was associated with significant increase in reactive oxygen species (ROS) and decrease in oxidized to reduced gluthatione ratio in RC6 cell line indicating a reduced level of oxidative stress. The mRNA expression levels of manganese superoxid dismutase (MnSOD), inducible nitric oxide synthase (iNOS) and gluthatione peroxidase (GPx) were increased while hypoxia-inducible factor 1-α (HIF-1α) was decreased in RC6 compared to C6 cells. This was in line with obtained changes in ROS content and increased antioxidative capacity of RC6 cells. Importantly, RC6 cells demonstrated collateral sensitivity to doxorubicin (DOX). The analysis of this phenomenon revealed increased accumulation of DOX in RC6 cells due to their adaptation to high ROS content and acidification of cytoplasm. In conclusion, newly established RC6 rat glioma cell line could be used as a starting material for the development of allogenic animal model and preclinical evaluation of new antiglioma agents. Collateral sensitivity to DOX obtained after BCNU treatment may prompt new studies aimed to find efficient delivery of DOX to the glioma site in brain.

Changes in mitochondria fusion protein-2 hepatic expression in conditions of liver cirrhosis and acute on chronic liver failure

To determine the differential protein and mRNA expressions of mitochondria fusion protein-2 (Mfn2) in hepatic tissues in conditions of cirrhosis and acute on chronic liver failure using rat model systems,and to determine the correlative effects on production of adenosine triphosphate (ATP) and reactive oxygen species (ROS). A liver cirrhotic rat model (LC rats) was established by intraperitoneal injection of carbon tetrachloride (CCl4,in vegetable oil),and these mice were subsequently used (10 weeks later) to establish the acute on chronic liver failure rat model (LF rats) by injecting lipopolysaccharide and D-amino-galactose.Control groups (normal controls,NC rats) were established for each model by intraperitoneal injection of vegetable oil only.Protein expression of Mfn2 in liver was quantified by western blotting with fluorescence densitometry and immunofluorescence staining,and mRNA expression was measured by real-time fluorescence quantitative PCR.ROS levels in liver were measured by fluorescence spectrophotometry,and ATP content was measured by bioluminescence assay.Significance of inter-group differences was assessed by one-way ANOVA,and correlations were determined using bivariate statistical modeling. Mfn2 protein expression was significantly lower in the liver tissues from modeled rats than that from the control rats (LC:0.051+/-0.004 and LF:0.037+/-0.007 vs.NC:0.254+/-0.008;F=444.98,P less than 0.05).The mRNA expression followed the same trend of lower expression (LC:21.21+/-0.93 and LF:24.35+/-0.85 vs.NC:19.09+/-0.69; F=66.941,P less than 0.05).The ATP content in liver tissues was also significantly lower in the modeled rats (LC:2.07+/-0.05 mol/L and LF:1.81+/-0.11 mol/L vs.NC:3.24+/-0.08 mol/L; F =574.21,P less than 0.05).Lower Mfn2 expression was correlated with lower ATP content (r =0.982) and higher ROS content (r =0.803). Reduced Mfn2 expression in liver tissue may cause a decrease in ATP synthesis and increase in ROS generation,thereby disrupting metabolism and increasing oxidative stress in the liver under conditions of cirrhosis and liver failure.

Cotton GhMPK6a negatively regulates osmotic tolerance and bacterial infection in transgenic Nicotiana benthamiana, and plays a pivotal role in development

Mitogen-activated protein kinase (MAPK) cascades play important roles in the perception of external signals and the generation of suitable responses. Cotton (Gossypium hirsutum) is an important fibre-producing and oil-producing crop worldwide. However, few MAPKs and their interaction partners have been functionally characterized in cotton. In the present study, the group A MAPK G. hirsutum (Gh)MPK6a was identified and characterized. GhMPK6a expression can be induced through multiple defence-related signal molecules and abiotic and biotic stresses. The ectopic expression of GhMPK6a in Nicotiana benthamiana reduced drought and salt tolerance, with elevated malondialdehyde content, higher reactive oxygen species content and lower abscisic acid content than in wild-type plants. Moreover, plants overexpressing GhMPK6a were sensitive to the bacterial pathogen Ralstonia solanacearum. Histochemical analysis of β-glucuronidase activity revealed that GhMPK6a showed tissue-specific expression during postgermination development, mixed bud differentiation, and pollination. Most importantly, GhMPK6a interacts with the upstream MAPK kinase GhMKK4, as shown by the use of yeast two-hybrid and bimolecular fluorescence complementation systems, compensating for a deficiency of MAPK interaction partners in cotton crops. Taken together, these results suggest that GhMPK6a negatively regulates osmotic stress and bacterial infection, and plays an important role in developmental processes. These results provide useful information for elucidating the roles of MAPK cascades in cotton crops. GhMPK6a physically interacts with GhMPK4 by two hybrid (View interaction) GhMPK6a and GhMPK4 physically interact by bimolecular fluorescence complementation (View interaction).

Trans-generational impact of cerium oxide nanoparticles on tomato plants

Cerium oxide nanoparticles (CeO2-NPs) are increasingly used in polishing, engine enhancement agents and many other products. Even though the acute toxicity of CeO2-NPs to plants has been investigated, the long-term effects of CeO2-NPs in the environment are still unknown. The main objective of this study was to investigate whether the treatment of tomato plants with relatively low concentrations of CeO2-NPs (10 mg L(-1)) through their lifecycle would affect the seed quality and the development of second generation seedlings. The results indicated that second generation seedlings grown from seeds collected from treated parent plants with CeO2-NPs (treated second generation seedlings) were generally smaller and weaker, as indicated by their smaller biomass, lower water transpiration and slightly higher reactive oxygen species content. An interesting phenomenon noticed in the study was that the second generation seedlings grown from treated seeds developed extensive root hairs compared with the control second generation seedlings (seedlings grown from seeds collected from untreated parent plants) regardless of the treatment. Treated second generation seedlings also accumulate a higher amount of ceria than control second generation seedlings under the same treatment conditions even though such differences are not statistically significant.

Mdm2 Is Required for Survival of Hematopoietic Stem Cells/Progenitors via Dampening of ROS-Induced p53 Activity

Mdm2 is an E3 ubiquitin ligase that targets p53 for degradation. p53(515C) (encoding p53R172P) is a hypomorphic allele of p53 that rescues the embryonic lethality of Mdm2(-/-) mice. Mdm2(-/-) p53(515C/515C) mice, however, die by postnatal day 13 resulting from hematopoietic failure. Hematopoietic stem cells and progenitors of Mdm2(-/-) p53(515C/515C) mice were normal in fetal livers but were depleted in postnatal bone marrows. After birth, these mice had elevated reactive oxygen species (ROS) thus activating p53R172P. In the absence of Mdm2, stable p53R172P induced ROS and cell cycle arrest, senescence, and cell death in the hematopoietic compartment. This phenotype was partially rescued with antioxidant treatment and upon culturing of hematopoietic cells in methycellulose at 3% oxygen. p16 was also stabilized because of ROS, and its loss increased cell cycling and partially rescued hematopoiesis and survival. Thus, Mdm2 is required to control ROS-induced p53 levels for sustainable hematopoiesis.