Growth and yield of groundnut (Arachis hypogaea L.) plants in soil inoculated with Aspergillus niger Van Tieghem

Aspergillus niger, a soil-borne fungus is a causative agent of hypocotyl malformations in infected groundnut (Arachis hypogaea L.) plants, but its effect on yield is unknown. This study sought to determine its effect on growth and yield. Seeds of Chinese and JL45 varieties were sown in soil inoculated with A. niger. Fresh and dry weights of the shoots and roots were taken at 10-day intervals. Nodule count was done at 30 days after emergence and subsequently at 10-day intervals. Pods of 20 plants each from inoculated and uninoculated soils were harvested. Growth was suppressed in plants grown on A. niger inoculated soil. Eight-day old plants grown in inoculated soil developed curvatures on their hypocotyls. Nodulation was suppressed (p < 0.05) in plants grown in inoculated soil. Although growth was suppressed in plants grown on inoculated soil, yield of both varieties of groundnut was not affected.     

Induction of Systemic Acquired Resistance in Arachis hypogaea L. by Sclerotium rolfsii Derived Elicitors

Plants evolve a strategy to survive the attacks of potential pathogens by inducing the microbial signal molecules. In this study, plant defence responses were induced in four different varieties of Arachis hypogaea (J‐11, GG‐20, TG‐26 and TPG41) using the fungal components of Sclerotium rolfsii in the form of fungal culture filtrate (FCF) and mycelial cell wall (MCW), and the levels of defence‐related signal molecule salicylic acid (SA), marker enzymes such as peroxidase (POX), phenylalanine ammonia lyase (PAL), β‐1,3‐glucanase and lignin were determined. There was a substantial fold increase in POX, PAL, SA, β‐1,3‐glucanase and lignin content in FCF‐ and MCW‐treated plants of all varieties of groundnut when compared to that of control plants. The enzyme activities were much higher in FCF‐treated plants than in MCW‐treated plants. The increase in fold activity of enzymes and signal molecule varied between different varieties. These results indicate that the use of fungal components (FCF and MCW) had successfully induced systemic resistance in the four different varieties of groundnut plants against Sclerotium rolfsii.     

Biological control of soil-pathogenic nematodes infecting mungbean using Pseudomonas fluorescens

The addition of organic matters to soil has been explored as an alternative means of nematode control under field conditions. Several oil-seed cakes of neem (Azadirachta indica), castor (Ricinus communis), groundnut (Arachis hypogeae), linseed (Linum usitatissimum) and sunflower (Helianthus annuus) were found to be highly effective in reducing the multiplication of soil-pathogenic nematodes Meloidogyne incognita, Rotylenchulus reniformis, Tylenchorhynchus brassicae, etc. The plant growth parameters such as plant weight, per cent pollen fertility, number of pods per plant, root-nodulation and chlorophyll content of mungbean increased significantly. The multiplication rate of nematodes and number of root-galls were less in the presence of Pseudomonas fluorescens as compared to its absence. Damage caused by the nematodes was further reduced when P. fluorescens was added along with the oil-seed cakes. Neem cake was found most effective in combination with P. fluorescens.     

Differential susceptibility of two varieties of cowpea (Vigna unguiculata (L) Walp) to phosphorus-induced zinc deficiency

Summary In a soil pot culture experiment conducted under greenhouse conditions, two varieties of cowpea (Vigna unguiculata (L) Walp) were found to be differentially susceptible to phosphorus-induced zinc deficiency. Although phosphate application, in the absence of applied zinc, caused growth disorder in both the varieties, the symptoms were more virulent, appeared early and were induced with both 25 and 75 ppm phosphorus in variety HFC-42-1, as against FOS-1, in which case zinc deficiency symptoms appeared under P₇₅Zn₀ treatment only. Contrary to the expectation the more susceptible variety was found to have higher zinc concentration in its shoots and thus the differential susceptibility of these varieties of cowpea could not be related to their zinc absorption characteristics. High phosphate application caused efflux of zinc during 16–32 day growth in both the varieties. However, higher sensitivity to phosphorus-induced zinc disorder in variety HFC-42-1, appeared to be due to nearly 2 times higher P concentration in this variety as against FOS-1. The results indicate that variety HFC-42-1 has a greater need for zinc because of its inherent capacity to accumulate more phosphorus. Dry matter yield and phosphorus and zinc content in the varieties were also differentially affected by the applications of both phosphate as well as zinc. re]19760504     

Biosorption of zinc by fungal mycelial wastes

Summary Waste mycelia from several industrial fermentation plants (Aspergillus niger, Penicillium chrysogenum, Claviceps paspali) were used as a biosorbent for zinc ions from aqueous environments, both batchwise as well as in a column mode. With all mycelia testes, biosorption per biomass dry weight was a function of pH (increasing with increasing pH between 1.0 and 9.0), biomass concentration (decreasing at high biomass concentrations) and the zinc concentration. Under optimized conditions, A. niger and C. paspali were superior to P. chrysogenum. Treatment of A. niger biomass with NaOH further increased its biosorbent capacity. Desorption of biosorbed zinc was achieved by elution with 0.1 m HCl, best results being obtained with NaOH-treated A. niger. Such treatment did not affect the capacity for biosorption in repeated experiments. NaOH-treated A. niger mycelia were also successfully used in removal of zinc from polluted waters in Austria, thereby showing that the simultaneous presence of other naturally occurring ions does not affect biosorption. Offprint requests to: C. P. Kubicek     

The inhibition kinetics of yeast glutathione reductase by some metal ions

Glutathione reductase (GR, type IV, Baker's yeast, E.C 1.6.4.2) is a flavoprotein that catalyzes the NADPH-dependent reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). In this study some metal ions have been tested on GR; lithium, manganese, molybdate, aluminium, barium, zinc, calcium, cadmium and nickel. Cadmium, nickel and calcium showed a good to moderate inhibitory effect on yeast GR. GR is inhibited non-competitively by Zn^{2 +} (up to 2 mM) and activated above this concentration. Ca^{2 +} inhibition was non-competitive with respect to GSSG and uncompetitive with respect to NADPH. Nickel inhibition was competitive with respect to GSSG and uncompetitive with respect to NADPH. The inhibition constants for these metals on GR were determined. The chelating agent EDTA recovered 90% of the GR activity inhibited by these metals.     

Control of the NADPH supply and GSH recycling for oxidative stress management in hepatoma and liver mitochondria

To unveil what controls mitochondrial ROS detoxification, the NADPH supply and GSH/GSSG recycling for oxidative stress management were analyzed in cancer and non-cancer mitochondria. Therefore, proteomic and kinetomic analyses were carried out of the mitochondrial (i) NADPH producing and (ii) GSH/GSSG recycling enzymes associated to oxidative stress management. The protein contents of the eight enzymes analyzed were similar or even higher in AS-30D rat hepatoma mitochondria (HepM) than in rat liver (RLM) and rat heart (RHM) mitochondria, suggesting that the NADPH/GSH/ROS pathway was fully functional in cancer mitochondria.The Vmax values of IDH-2 were much greater than those of GDH, TH and ME, suggesting that IDH-2 is the predominant NADPH producer in the three mitochondrial types; in fact, the GDH reverse reaction was favored. The Vmax values of GR and GPx were lower in HepM than in RLM, suggesting that the oxidative stress management is compromised in cancer mitochondria. The Km values of IDH-2, GR and GPx were all similar among the different mitochondrial types.Kinetic modeling revealed that the oxidative stress management was mainly controlled by GR, GPx and IDH. Modeling and experimentation also revealed that, due to their higher IDH-2 activity and lower GPx activity presumably by acetylation, HepM (i) showed higher steady-state NADPH levels; (ii) required greater peroxide concentrations to achieve reliable steady-state fluxes and metabolite concentration; and (iii) endured higher peroxide concentrations without collapsing their GSH/GSSG ratios. Then, to specifically prompt lower GSH/GSSG ratios under oxidative stress thus compromising cancer mitochondria functioning, GPx should be re-activated.     

Expression of fluorescent proteins in Lactobacillus rhamnosus to study host–microbe and microbe–microbe interactions

Probiotic Lactobacillus strains are widely used to benefit human and animal health, although the exact mechanisms behind their interactions with the host and the microbiota are largely unknown. Fluorescent tagging of live probiotic cells is an important tool to unravel their modes of action. In this study, the implementation of different heterologously expressed fluorescent proteins for the labelling of the model probiotic strains Lactobacillus rhamnosusGG (gastrointestinal) and Lactobacillus rhamnosusGR‐1 (vaginal) was explored. Heterologous expression of mTagBFP2 and mCherry resulted in long‐lasting fluorescence of L. rhamnosusGG and GR‐1 cells, using the nisin‐controlled expression (NICE) system. These novel fluorescent strains were then used to study in vitro aspects of their microbe–microbe and microbe–host interactions. Lactobacillus rhamnosusGG and L. rhamnosusGR‐1 expressing mTagBFP2 and mCherry could be visualized in mixed‐species biofilms, where they inhibited biofilm formation by Salmonella Typhimurium–gfpmut3 expressing the green fluorescent protein. Likewise, fluorescent L. rhamnosusGG and L. rhamnosusGR‐1 were implemented for the visualization of their adhesion patterns to intestinal epithelial cell cultures. The fluorescent L. rhamnosus strains developed in this study can therefore serve as novel tools for the study of probiotic interactions with their environment.     

Effect of intercropping maize and soybeans on Striga hermonthica parasitism and yield of maize

Striga hermonthica a major biotic constraint to cereal production can be controlled by trap crops. Soybean cultivars vary in ability to stimulate suicidal germination of the weed. An experiment was conducted to select soybean (Glycine max) varieties with the ability to stimulate germination of S. hermonthica seeds. Experiments were conducted with strigol Nijmegen 1® (GR 24), a synthetic stimulant, as a check. In the pot and field experiments, maize (variety WH507) was intercropped with soybeans. Variation occurred among soybean varieties in inducing germination of S. hermonthica. The relative germination induction by soybean varieties ranged from 8% to 66% compared to 70% for synthetic stimulant check. Varieties TGX1448-2E, Tgm 1576, TGX1876-4E and Tgm 1039 had the highest relative germination. Soybean varieties TGX 1831-32E, Tgm944, Tgm 1419 and Namsoy4m had high stimulation but low attachment. Intercropping maize with soybeans in the field led to a low S. hermonthica count and high maize yield.     

Expression of a glutathione reductase from <Emphasis Type="Italic">Brassica rapa</Emphasis> subsp. <Emphasis Type="Italic">pekinensis</Emphasis> enhanced cellular redox homeostasis by modulating antioxidant proteins in <Emphasis Type="Italic">Escherichia coli</Emphasis>

Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semiquantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to H₂O₂, menadione, and heavy metal (CdCl₂, ZnCl₂ and AlCl₂)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to H₂O₂ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.     

The complex roles of NADPH oxidases in fungal infection

NADPH oxidases play key roles in immunity and inflammation that go beyond the production of microbicidal reactive oxygen species (ROS). The past decade has brought a new appreciation for the diversity of roles played by ROS in signalling associated with inflammation and immunity. NADPH oxidase activity affects disease outcome during infections by human pathogenic fungi, an important group of emerging and opportunistic pathogens that includes Candida, Aspergillus and Cryptococcus species. Here we review how alternative roles of NADPH oxidase activity impact fungal infection and how ROS signalling affects fungal physiology. Particular attention is paid to roles for NADPH oxidase in immune migration, immunoregulation in pulmonary infection, neutrophil extracellular trap formation, autophagy and inflammasome activity. These recent advances highlight the power and versatility of spatiotemporally controlled redox regulation in the context of infection, and point to a need to understand the molecular consequences of NADPH oxidase activity in the cell.     

Effect of cultural methods on Striga (Striga hermonthica (Del.)) Benth management and yield of cereals in the Savanna Zone of Nigeria: a review

Data were obtained from the research done in the Guinea Savanna (Zaria 11°11′N; 070 38′E) and Sudan Savanna (Maiduguri ?11°51′N; 13°15′E) regions of Nigeria, respectively on different cultural methods of Striga control and management. In the Guinea Savanna, trials on the effects of nitrogen on the response of resistant and susceptible upland rice varieties to Striga hermonthica infestation and the effect of resistant and susceptible varieties of maize and crop rotation on Striga infestation was carried out, while the effect of inter-cropping trap crop (Bambaranut) with resistant sorghum varieties on S. hermonthica was studied in the Sudan Savanna Zone of Nigeria. In the Guinea Savanna, it was observed that a combination of upland rice variety, Faro 40 and an application of 90 kg N/ha in the wet season and WAB 56-50 upland rice variety and 120 kg N/ha in the dry season, respectively reduced Striga infestation and produced maximum grain yield. Also, the growing of resistant variety of maize (Across 97ITZ comp. I-W) after 1 or 2 years' rotation with cowpea or soybean was observed not only to be effective in Striga control, but resulted in higher grain yield of maize. In the Sudan Savanna, the use of resistant varieties of sorghum, 1CSV1002 and 1CSV1007 intercropped with bambaranut significantly reduced Striga infestation, but the grain yield of the resistant varieties was low. From these studies, Faro 40 with 90 kg N/ha application rate and WAB56–50 with 120 kg N/ha were suitable for the management of Striga and for higher grain yield of upland rice in both wet and dry seasons, respectively, while Across 97ITZ comp. I-W, resistant maize variety and 1 or 2 years rotation with cowpea or soybean were also the best for the management of S. hermonthica and for higher maize yield in the Guinea Savanna zone. Further research needs to be carried out in the Sudan Savanna to select a high yielding resistant variety of sorghum which when intercropped with bambaranut will not only control Striga infestation but will also give high grain yield.     

Down-regulation of neprilysin (EC3.4.24.11) expression in vascular endothelial cells by laminar shear stress involves NADPH oxidase-dependent ROS production

Neprilysin (NEP, neutral endopeptidase, EC3.4.24.11), a zinc metallopeptidase expressed on the surface of endothelial cells, influences vascular homeostasis primarily through regulated inactivation of natriuretic peptides and bradykinin. Earlier in vivo studies reporting on the anti-atherosclerotic effects of NEP inhibition and on the atheroprotective effects of flow-associated laminar shear stress (LSS) have lead us to hypothesize that the latter hemodynamic stimulus may serve to down-regulate NEP levels within the vascular endothelium. To address this hypothesis, we have undertaken an investigation of the effects of LSS on NEP expression in vitro in bovine aortic endothelial cells (BAECs), coupled with an examination of the signalling mechanism putatively mediating these effects. BAECs were exposed to physiological levels of LSS (10 dynes/cm², 24 h) and harvested for analysis of NEP expression using real-time PCR, Western blotting, and immunocytochemistry. Relative to unsheared controls, NEP mRNA and protein were substantially down-regulated by LSS (≥50%), events which could be prevented by treatment of BAECs with either N-acetylcysteine, superoxide dismutase, or catalase, implicating reactive oxygen species (ROS) involvement. Employing pharmacological and molecular inhibition strategies, the signal transduction pathway mediating shear-dependent NEP suppression was also examined, and roles implicated for Gβγ, Rac1, and NADPH oxidase activation in these events. Treatment of static BAECs with angiotensin-II, a potent stimulus for NADPH oxidase activation, mimicked the suppressive effects of shear on NEP expression, further supporting a role for NADPH oxidase-dependent ROS production. Interestingly, inhibition of receptor tyrosine kinase signalling had no effect. In conclusion, we confirm for the first time that NEP expression is down-regulated in vascular endothelial cells by physiological laminar shear, possibly via a mechanotransduction mechanism involving NADPH oxidase-induced ROS production.     

Purification and Characterisation of Rat Kidney Glutathione Reductase

Glutathione reductase [GR, E.C.1.8.1.7] catalyses NADPH dependent reduction of glutathione disulfide (GSSG) to reduced glutathione (GSH). Thus, it is the crucial enzyme to maintain high [GSH]/[GSSG] ratio and physiological redox status in cells. Kidney and liver tissues were considered as a rich source of GR. In this study, rat kidney GR was purified and some of its properties were investigated. The enzyme was purified 2,356 fold with a yield of 16% by using heat-denaturation and Sephadex G25 gel filtration, 2′,5′-ADP Agarose 4B, PBE94 column chromatographies. The purified enzyme had a specific activity (Vm) of 250 U/mg protein and the ratio of absorbances at wavelengths of A ₂₇₃/A _463, A ₂₈₀/A ₄₆₀, A ₃₆₅/A ₄₆₀, and A ₃₇₉/A ₄₆₃, were 7.1, 6.8, 1.2 and 1.0, respectively. Each mol of GR subunit bound 0.97 mol of FAD. NADH was used as a coenzyme by rat kidney GR but with a lower efficiency (32.7%) than NADPH. Its subunit molecular weight was estimated as 53 kDa. An optimum pH of 6.5 and optimum temperature of 65 °C were found for rat kidney GR. Its activation energy (Ea) and temperature coefficient (Q₁₀) were calculated as 7.02 kcal/mol and 1.42, respectively. The Km_(NADPH) and kcat/Km _(NADPH) values were found to be 15.3 ± 1.4 μM and 1.68 × 10⁷ M⁻¹ s⁻¹ for the concentration range of 10-200 μM NADPH and when GSSG is the variable substrate, the Km_(GSSG) and the kcat/Km_(GSSG) values of 53.1 ± 3.4 μM and 4.85 × 10⁶ M⁻¹ s⁻¹ were calculated for the concentration range of 20–1,200 μM GSSG.     

Inhibition of purified bovine liver glutathione reductase with some metal ions

Glutathione reductase (GR; E.C. 1.6.4.2) is a flavoprotein that catalyzes the NADPH-dependent reduction of oxidized glutathione (GSSG). In this study we tested the effects of Al³⁺, Ba²⁺, Ca²⁺, Li⁺, Mn²⁺, Mo⁶⁺, Cd²⁺, Ni²⁺, and Zn²⁺ on purified bovine liver GR. In a range of 10?μM–10?mM concentrations, Al³⁺, Ba²⁺, Li⁺, Mn²⁺, and Mo⁶⁺, and Ca²⁺ at 5?μM–1.25?mM, had no effect on bovine liver GR. Cadmium (Cd²⁺), nickel (Ni²⁺), and zinc (Zn²⁺) showed inhibitory effects on this enzyme. The obtained IC₅₀ values of Cd²⁺, Ni²⁺, and Zn²⁺ were 0.08, 0.8, and 1?mM, respectively. Cd²⁺ inhibition was non-competitive with respect to both GSSG (Ki_GSSG 0.221?±?0.02?mM) and NADPH (Ki_NADPH 0.113?±?0.008?mM). Ni²⁺ inhibition was non-competitive with respect to GSSG (Ki_GSSG 0.313?±?0.01?mM) and uncompetitive with respect to NADPH (Ki_NADPH 0.932?±?0.03?mM). The effect of Zn²⁺ on GR activity was consistent with a non-competitive inhibition pattern when the varied substrates were GSSG (Ki_GSSG 0.320?±?0.018?mM) and NADPH (Ki_NADPH 0.761?±?0.04?mM), respectively.     

Identification of Critical Stage for Disease Development and Biocontrol of Alternaria Blight of Indian Mustard (Brassica juncea)

Fungicides mancozeb and carbendazim caused 100% reduction in mycelial growth of Alternaria brassicae over control in vitro while 1% (w/v) aqueous bulb extract of Allium sativum and leaf extract of Acacia nilotica caused significant reductions. In dual culture, GR isolate of Trichoderma viride performed the best among the test isolates of Trichoderma, causing 81%, 82% reduction in mycelial growth of A. brassicae over control. Performance of isolates SI‐2, P and SI‐1 of T. viride were at par (P < 0.01) with that of GR isolate. Spraying of A. brassicae at different ages of the mustard host plant identified 75 days after sowing (d.a.s.) as the most critical age of the mustard plant for development of Alternaria blight severity on the crop with 45 d.a.s. being the next most important one. Mancozeb was the best among all the treatments, resulting in the lowest disease severity on leaves of mustard at both Sewar and Ludhavai as also the lowest A‐value (area under disease progress curve). Performance of bulb extract of A. sativum in checking the disease severity on leaves and pods was at par (P < 0.05) with mancozeb. The GR isolate of T. viride was at par with mancozeb in checking blight severity on mustard leaves at Sewar while performance of the bioagent was significantly (P < 0.05) inferior to the chemical fungicide at Ludhavai. Performance of the bioagent isolate GR of T. viride in checking the disease severity on pods was at par (P < 0.05) with mancozeb at both Sewar and Ludhavai, the treatment recording the lowest A‐value on pods. While application of bulb extract of A. sativum resulted in highest seed yield at Sewar in 2001–2002, the bioagent isolate GR of T. viride did so at Ludhavai, both the treatments being at par (P < 0.05) with mancozeb and significantly higher than control. Application of bulb extract of A. sativum at 45 and 75 d.a.s. resulted in lowest blight severity on leaves and pods as also in highest seed yield among the different single and combination of treatments. Although disease severity in the treatment was at par (P < 0.05) with that in mancozeb, application of the plant extract at the two stages of crop growth resulted in significantly higher seed yield compared with the two applications of the chemical fungicide. However, application of the treatments singly only at 75 d.a.s., GR isolate of T. viride at 45 and 75 d.a.s., A. sativum 45 d.a.s. + T. viride 75 d.a.s., and T. viride 45 d.a.s. + A. sativum 75 d.a.s. resulted in seed yield at par (P < 0.05) with application of bulb extract of A. sativum at 45 and 75 d.a.s.     

Bulge Formation in Escherichia coli Induced by l-Arabinose under Hypertonic Conditions with Sodium Chloride

A pseudotropine-forming tropinone reductase was extracted from root cultures of Hyoscyamus niger that produce the tropane alkaloids hyoscyamine and scopolamine. The enzyme stereospecifically reduces tropinone to pseudotropine, oxidizing NADPH. It has an approximate molecular weight of 84,000 and a pH optimum between 5.8 and 6.25. The Km value for tropinone is 35.1 μmol/l and for NADPH 21.1 μmol/l. Substrate specificity was tested for NADPH and several tropinone analogues.     

Contribution of glucocorticoid-mineralocorticoid receptor pathway on the obesity-related adipocyte dysfunction

AimsMineralocorticoid receptor (MR) blockade ameliorated insulin resistance with improvements in adipocytokine dysregulation, inflammation, and excess of reactive oxygen species (ROS) in obese adipose tissue and adipocytes, but its mechanism has not been clarified. The 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), producing active glucocorticoids, is highly expressed in adipocytes and glucocorticoids bind to MR with higher affinity than to glucocorticoid receptor (GR). We investigated whether glucocorticoids effect on adipocytokines and ROS through MR in adipocytes. In addition, fat distributions of MR and GR were investigated in human subjects.Methods and ResultsCorticoid receptors and their target genes were examined in adipose tissue of obese db/db mice. 3T3-L1 adipocytes were treated with glucocorticoids, H₂O₂, MR antagonist eplerenone (EP), GR antagonist RU486 (RU), MR-siRNA, and/or N-acetylcysteine. Human adipose tissues were obtained from seven patients who underwent abdominal surgery. The mRNA levels of MR and its target gene were higher in db/db mice than in control db/m + mice. In 3T3-L1 adipocytes, glucocorticoids, similar to H₂O₂, caused the dysregulation of mRNA levels of various genes related to adipocytokines and the increase of intracellular ROS. Such changes were rectified by MR blockade, not by GR antagonist. In human fat, MR mRNA level was increased in parallel with the increase of body mass index (BMI) and its increase was more significant in visceral fat, while there were no apparent correlations of GR mRNA level to BMI or fat distribution.ConclusionGlucocorticoid-MR pathway may contribute to the obesity-related adipocytokine dysregulation and adipose ROS.     

Up‐regulation of glycolysis promotes the stemness and EMT phenotypes in gemcitabine‐resistant pancreatic cancer cells

Cancer stem cells (CSCs) and epithelial–mesenchymal transition (EMT)‐type cells are considered as underlying causes of chemoresistance, tumour recurrence and metastasis in pancreatic cancer. We aimed to describe the mechanisms – particularly glycolysis – involved in the regulation of the CSC and EMT phenotypes. We used a gemcitabine‐resistant (GR) Patu8988 cell line, which exhibited clear CSC and EMT phenotypes and showed reliance on glycolysis. Inhibition of glycolysis using 2‐deoxy‐D‐glucose (2‐DG) significantly enhanced the cytotoxicity of gemcitabine and inhibited the CSC and EMT phenotypes in GR cells both in vitro and in vivo. Intriguingly, the use of the reactive oxygen species (ROS) scavenger N‐acetylcysteine (NAC) restored the CSC and EMT phenotypes. H₂O₂ produced changes similar to those of 2‐DG, indicating that ROS were involved in the acquired cancer stemness and EMT phenotypes of GR cells. Moreover, doublecortin‐like kinase 1 (DCLK1), a pancreatic CSC marker, was highly expressed and regulated the stemness and EMT phenotypes in GR cell. Both 2‐DG and H₂O₂ treatment suppressed DCLK1 expression, which was also rescued by NAC. Together, these findings revealed that glycolysis promotes the expression of DCLK1 and maintains the CSC and EMT phenotypes via maintenance of low ROS levels in chemoresistant GR cells. The glycolysis‐ROS‐DCLK1 pathway may be potential targets for reversing the malignant behaviour of pancreatic cancer.