Soluble urate-induced effects on cytokine production in vitro - Assessment of methodologies and cell types.

BACKGROUND: Hyperuricemia has been shown to be an inducer of pro-inflammatory mediators by human primary monocytes. To study the deleterious effects of hyperuricemia, a reliable and stable in vitro model using soluble urate is needed. One recent report showed different urate-dissolving methods resulted in either pro-inflammatory or anti-inflammatory properties. The aim of this study was to compare the effect of two methods of dissolving urate on both primary human peripheral blood mononuclear cells (PBMCs) and THP-1 cells. The two methods tested were 'pre-warming' and 'dissolving with NaOH'. METHODS: Primary human PBMCs and THP-1 cells were exposed to urate solutions, prepared using the two methodologies: pre-warming and dissolving with NaOH. Afterwards, cells were stimulated with various stimuli, followed by the measurement of the inflammatory mediators IL-1ß, IL-6, IL-1Ra, TNF, IL-8, and MCP-1. RESULTS: In PBMCs, we observed an overall pro-inflammatory effect of urate, both in the pre-warming and the NaOH dissolving method. A similar pro-inflammatory effect was seen in THP-1 cells for both dissolving methods after restimulation. However, THP-1 cells exhibited pro-inflammatory profile with exposure to urate alone without restimulation. We did not find MSU crystals in our cellular assays. CONCLUSIONS: Overall, the urate dissolving methods do not have critical impact on its inflammatory properties. Soluble urate prepared using either of the two methods showed mostly pro-inflammatory effects on human primary PBMCs and monocytic cell line THP-1. However, human primary PBMCs and the THP-1 differ in their response to soluble urate without restimulation.

A robust soft sensor based on artificial neural network for monitoring microbial lipid fermentation processes using Yarrowia lipolytica.

Microbial oils produced by Yarrowia lipolytica offer an environmentally friendly and sustainable alternative to petroleum as well as traditional lipids from animals and plants. The accurate measurement of fermentation parameters, including the substrate concentration, dry cell weight, and lipid accumulation, is the foundation of process control, which is indispensable for industrial lipid production. However, it remains a great challenge to measure the complex parameters online during the lipid fermentation process, which is nonlinear, multivariate, and characterized by strong coupling. As a type of AI technology, the artificial neural network model is a powerful tool for handling extremely complex problems, and it can be employed to develop a soft sensor to monitor the microbial lipid fermentation process of Y. lipolytica. In this study, we first analyzed and emphasized the volume of sodium hydroxide and dissolved oxygen concentration as central parameters of the fermentation process. Then, a soft sensor based on a four-input artificial neural network model was developed, in which the input variables were fermentation time, dissolved oxygen concentration, initial glucose concentration, and additional volume of sodium hydroxide. This provides the possibility of online monitoring of dry cell weight, glucose concentration, and lipid production with high accuracy, which can be extended to similar fermentation processes characterized by the addition of bases or acids, as well as changes of the dissolved oxygen concentration.

Comparing the effects of gamma ray and alkaline treatments of sodium hydroxide and calcium oxide on chemical composition, ruminal degradation kinetics and crystallinity degree of soybean straw.

This research was conducted to compare the effects of gamma ray (GR) at doses of 100 and 150 kGy, alkaline treatments of sodium hydroxide (NaOH) and calcium oxide (CaO) at concentrations of 50 and 160 g/kg of dry matter (DM) respectively, and their combined use on chemical composition, ruminal degradation kinetics and crystallinity degree of soybean straw (SBS). After processing, the chemical composition of the samples was determined using the AOAC standard methods. Ruminal degradability trial was carried out by the nylon bag technique. Crystallinity degree of the samples was investigated through X-ray diffraction (XRD) technique. Using alkaline compounds in processing of SBS significantly increased the Ash amount (P < 0.0001). The greatest value was observed in CaO and CaO + GR treatments (100 and 150 kGy). The crude fiber (CF) of samples was reduced due to the effect of gamma irradiation, alkaline compounds, and their combination (P < 0.0001). Samples irradiated with 150 kGy dose of GR had the least amount of CF. The processing increased the effective rumen degradability (ERD) of DM for SBS at rumen outflow rates of 0.02, 0.05, and 0.08/h (P < 0.0001). The greatest amounts at the mentioned outflow rates were observed in GR (100, and 150 kGy), and NaOH + GR (100 and 150 kGy) treatments. Different treatments reduced the crystallinity degree of SBS compared to the control group. The least crystallinity percentage was observed in GR (150 kGy) treated samples. Totally, gamma irradiation, alkaline treatments and their combined use improved the nutritional value of SBS. In this respect, gamma irradiation was more effective than alkaline compounds.

Effects of segmented aerobic and anaerobic fermentation assisted with chemical treatment on comprehensive properties and composition of wheat straw.

Traditional aerobic composting used for straw treatment shows limited regulation effects and unstable properties, and it is necessary to introduce some co-processing methods to optimize its performance. Herein, segmented aerobic/anaerobic fermentation, combined with chemical treatment with wood vinegar/NaOH, was used to treat wheat straw. The results showed that anaerobic fermentation when used as the first stage could stabilize the wheat straw pH between 5.19 and 6.13 and improve nutrient contents. All treatments had greater effects on substrate aeration porosities (range of 14%) than on total porosity (range of 6%), and the water-holding porosities were improved to a greater extent by NaOH than by wood vinegar. The hemicellulose degradation rate of aerobic-anaerobic treatment was higher than that achieved with anaerobic-aerobic treatment, while the latter method was more effective at removing the neutral detergent-soluble as well as remaining organic matter, which was generated due to a higher KCl content in the ash.

CDK1 promotes the proliferation of melanocytes in Rex rabbits.

BACKGROUND: The fur color constitutes one of the most important economic characteristics of fur animals and is determined by the content of melanin. A previous study has shown that the cyclin-dependent kinase 1 (CDK1) is a member of the protein kinase family, involved in forming the color of the fur in Rex rabbits. However, its effect on the melanocytes remains unclear. OBJECTIVE: This study aimed to provide evidence for the role of CDK1 in melanogenesis. METHODS: This study measured the expression of CDK1 in Rex rabbit skins of six coat colors using qRT-PCR. The CDK1-mediated regulation of the pigmentation-related genes and cyclin-dependent kinases were analyzed. The melanin content, proliferation, and apoptosis of the melanocytes were analyzed using the NaOH, CCK8, and Annexin V-FITC methods. RESULTS: The CDK1 expression in the skin of the rex rabbits with different coat colors was found to be regular, and the expression level was found to be the highest in the skin of the black rex rabbits (P < 0.05). The overexpression/knockdown of CDK1 was found to significantly increase/decrease the melanin content in the melanocytes (P < 0.01). Besides, CDK1 was found to significantly promote the proliferation of the melanocyte and inhibit apoptosis (P < 0.01). Furthermore, the overexpression of CDK1 was found to significantly affect the expression of the other melanin-related genes like TYR, PMEL, DCT, as well as the mRNA expression of the cyclin-dependent kinases CDK4, CDK6, CDK8, CCNB1. CONCLUSIONS: The results indicated that CDK1 can serve as a key gene regulating melanogenesis, melanocyte proliferation, and apoptosis, providing a new theoretical basis for studying the mechanism by which the different colors of the fur evolve in mammals.

Towards a broader view of the metabolome: untargeted profiling of soluble and bound polyphenols in plants.

Phenylalanine (Phe) is a central precursor for numerous secondary plant metabolites with a multitude of biological functions. Recent studies on the fungal disease Fusarium head blight in wheat showed numerous Phe-derived defence metabolites to be induced in the presence of the pathogen. These studies also suggest a partial incorporation of Phe-derived secondary metabolites into the cell wall. To broaden the view of the metabolome to bound Phe derivatives, an existing approach using 13C-labelled Phe as tracer was extended. The developed workflow consists of three successive extractions with an acidified acetonitrile-methanol-water mixture to remove the soluble plant metabolites, followed by cell wall hydrolysis with 4M aqueous NaOH, acidification with aqueous HCl, and liquid-liquid extraction of the hydrolysate with ethyl acetate. The untargeted screening of Phe-derived metabolites revealed 156 soluble compounds and 90 compounds in the hydrolysed samples including known cell wall constituents like ferulic acid, coumaric acid, and tricin. Forty-nine metabolites were found exclusively in the hydrolysate. The average cumulative extraction yield of the soluble metabolites was 99.6%, with a range of 91.8 to 100%. Repeatability coefficients of variation of the protocol ranged from 10.5 to 25.9%, with a median of 16.3%. To demonstrate the suitability of the proposed method for a typical metabolomics application, mock-treated and Fusarium graminearum-treated wheat samples were compared. The study revealed differences between the hydrolysates of the two sample types, confirming the differential incorporation of Phe-derived metabolites into the cell wall under infection conditions.

Triiodothyronine improves contractile recovery of human atrial trabeculae after hypoxia/reoxygenation.

BACKGROUND: In patients undergoing interventional or surgical coronary revascularization, subclinical hypothyroidism is common and associated with worse outcome, including the need for postoperative inotropic support. In isolated rat hearts with global ischemia/reperfusion, exogenous triiodothyronine (T3) reduces infarct size. Aim of this study was, to investigate whether or not exogenous T3 protects human myocardium from ischemia/reperfusion injury. METHODS: Right atrial trabeculae from patients undergoing routine coronary artery bypass grafting were isolated and transferred to Tyrode's buffer. Electrically initiated (1 Hz) contractile stress (mN/mm2) was recorded for 10 min at baseline (95% O2/ 5% CO2, glucose). Sixty min hypoxia were induced by changing buffer gas and increasing stimulation rate (95% N2/ 5% CO2, choline chloride, 3 Hz) before return to reoxygenation for 30 min. T3 (500 µg/l) vs. NaOH (solvent control) was administered A) throughout (n = 11 vs. n = 9) or B) only 15 min before and during reoxygenation (n = 12 vs. n = 13). Western blot analyses of established cardioprotective signaling proteins were performed. RESULTS: At baseline, contractile stress was comparable. T3 improved the cumulative recovery of contractile stress during reoxygenation from 41 ± 16 with NaOH to 55 ± 11% of baseline with T3, when given continuously in A or from 52 ± 13 with NaOH to 63 ± 11% of baseline with T3 when given just before and during reoxygenation in B. The ratio of mitochondrial complex I matrix arm to membrane NADH:ubiquinone oxidoreductase subunits (NDUF)V2 to NDUFA9 was reduced, reflecting increased complex I activity. CONCLUSION: T3 increases contractile recovery of human right atrial trabeculae from hypoxia/reoxygenation.

Chemical composition and in situ degradability of sugarcane tip hay subjected to alkaline treatment.

The present study aimed to evaluate the effect of alkaline treatments with urea, NaOH and Ca(OH)2 on chemical composition and in situ ruminal degradability of dry matter, crud protein and neutral detergent fiber of sugarcane tip hay. Samples were incubated in the rumen of three cannulated cattle for up to 72 hours in a split plot randomized block design. Ammoniation with 6% urea increased (p<0.05) the crude protein content by 13% and reduced the neutral detergent fiber and insoluble nitrogen content of the hay. When treated with the highest doses of the compounds, there was a high potential degradability of dry matter, crude protein and neutral detergent fiber, and a shorter neutral detergent fiber lag time. Ammoniation with urea promotes a reduction in the content of NDF, hemicellulose and insoluble nitrogen, with an increase in the content of CP in the hay, with emphasis for the level of 6% urea. The ruminal degradation of sugarcane tip hay increases with alkaline treatments using 6% urea or 3% NaOH, however, ammoniation with urea is indicated for the treatment of hay, as this is low cost and can be easily adopted by farmers in the semiarid region.

Topical Effects of N-Acetyl Cysteine and Doxycycline on Inflammatory and Angiogenic Factors in the Rat Model of Alkali-Burned Cornea.

The aim of this study was to analyze the single and combined effects of N-acetyl cysteine (NAC) and doxycycline (Dox) on the inflammatory and angiogenic factors in the rat model of alkali-burned cornea. Rats were treated with a single and combined 0.5% NAC and 12.5 µg/mL Dox eye drops and evaluated on days 3, 7, and 28. In the corneas of various groups, the activity of Catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) enzymes was assessed. The expression of inflammatory factors (TNF-α, Rel-a, and CXCL-1) and angiogenic factors (VEGF-a, MMP2, and MMP9) was measured using real-time polymerase chain reaction. The antioxidant enzyme activities decreased substantially 3 days after injury with sodium hydroxide (NaOH). NAC and combined NAC+ Dox topical treatments increased the SOD enzyme activity on day 28 (P < 0.05). The expression of TNF-α and Rel-a genes following single and combined treatment of NAC and Dox decreased significantly on days 7 and 28 (P < 0.05). The mRNA level of angiogenic factors and corneal neovascularization (CNV) level declined in NaOH-injured rats treated with Dox (P < 0.05). The topical treatment of Dox could attenuate inflammation and CNV complications. However, NAC treatment may not reduce the expression of angiogenic genes.

Mild alkaline pretreatment can achieve high hydrolytic and fermentation efficiencies for rice straw conversion to bioethanol.

Mild alkaline pretreatment was evaluated as a strategy for effective lignin removal and hydrolysis of rice straw. The pretreatment efficiency of different NaOH concentrations (0.5, 1.0, 1.5 or 2.0% w/w) was assessed. Rice straw (RS) pretreated with 1.5% NaOH achieved better sugar yield compared to other concentrations used. A cellulose conversion efficiency of 91% (45.84 mg/ml glucose release) was attained from 1.5% NaOH pretreated rice straw (PRS), whereas 1% NaOH pretreated rice straw yielded 35.10 mg/ml of glucose corresponding to a cellulose conversion efficiency of 73.81%. The ethanol production from 1% and 1.5% NaOH pretreated RS hydrolysates was similar at ∼3.3% (w/v), corresponding to a fermentation efficiency of 86%. The non-detoxified hydrolysate was fermented using the novel yeast strain Saccharomyces cerevisiae RPP-03O without any additional supplementation of nutrients.

Heat and Chemical Treatments Affect the Viability, Morphology, and Physiology of Staphylococcus aureus and Its Subsequent Antibody Labeling for Flow Cytometric Analysis.

The effects of heat and chemical treatments on Staphylococcus aureus viability and physiology and their subsequent effects on antibody binding ability and cell morphology were measured. Treatments included lethal and sublethal heat; exposure to organic acids, salt, and sodium hydroxide; and freeze-thawing. Strain-related differences in viability were noted depending on treatment and were reflected in changes in physiology as monitored by flow cytometry (FCM) using three different staining protocols: SYTO 9/propidium iodide (PI), DiOC2(3), or calcein acetoxymethyl ester (calcein-AM)/PI. Treatments that resulted in significant losses in viability as measured by plate counting were reflected better by the first two staining combinations, as intracellular calcein-AM uptake may have been impaired by certain treatments. FCM analysis using labeling by commercial anti-S. aureus antibodies indicated that differences in cell physiology as a result of treatments influenced immunofluorescence detection. The ratio of the mean fluorescence intensities of stained cells to those of unstained cells [MFI/MFI(us)] varied with treatment, five of these treatments, including freeze-thaw, citric acid, oxalic acid, NaCl, and NaOH treatments, resulted in significantly lower fluorescence values compared to controls.IMPORTANCE FCM data indicated that cells conventionally considered to be dead and which would not give rise to CFU in a plate count assay, e.g., cells heated to 80°C, were labeled by antibody staining. This finding suggests that without the inclusion of a live/dead discriminating dye, these cells would be erroneously detected as viable within an FCM assay. Reductions in antibody staining due to physicochemical treatment were strain related, reflecting the complexity of the phenomenon under study and illustrating that substantial validation of any new antibody detection-based method, including physiological staining and cell sorting, should be undertaken. Researchers should be aware of physicochemical treatments causing false-negative results: in this study, freeze-thawing severely reduced antibody binding without affecting the viability of a substantial percentage of cells. Scanning electron microscopy carried out on treated cells revealed a range of morphological changes resulting from physicochemical treatments which may have hindered antibody binding.

Alkaline and Alkaline-Oxidative Pretreatment and Hydrolysis of Herbaceous Biomass for Growth of Oleaginous Microbes.

This chapter describes methods for generation of hydrolysates amenable to conversion to microbial lipids from herbaceous lignocellulosic biomass utilizing either mild alkali pretreatment with NaOH or alkaline hydrogen peroxide pretreatment with NaOH and H2O2. This pretreatment is followed by enzymatic hydrolysis of the plant cell wall polysaccharides to yield hydrolysates. These hydrolysates are composed primarily of the monosaccharides glucose and xylose as well as acetate and phenolic monomers that may all serve as a source of renewable carbon to produce microbial lipids. Application of these mild pretreatment conditions minimizes the generation of inhibitors, enabling microbial cultivations to often be performed without the need for detoxification.

Effect of Supplementation of Nano Zinc Oxide on Nutrient Retention, Organ and Serum Minerals Profile, and Hepatic Metallothionein Gene Expression in Wister Albino Rats.

A study was conducted to validate the effects of nano form of zinc (NZn) on nutrient digestibility, zinc retention, organ and serum zinc profile, and hepatic metallothionein gene expression in Wistar albino rats (WAR). Nano zinc (NZn) was synthesized through chemical method, by using 0.45 M zinc nitrate [Zn(NO3)2.6H2O] and 0.9 M sodium hydroxide (NaOH). The NZn particle in its oxide form was characterized by TEM-EDAX and XRD, and found to be in nano range (below 100 nm. Zinc was supplemented to the Wistar albino rats (WAR) through synthetic semi-purified diet either without Zn, or as inorganic zinc (IZn; 25 mg/kg), or as synthesized NZn (25, 12.5, 6.25, 3.125 or 50 mg/kg DM) for 60 days. The zinc content was observed to be significantly (P < 0.05) higher in liver, bone, kidney, and serum due to NZn supplementation where NZn-50 had highest zinc content and control had the least, without affecting Fe, Mn, and Cu. NZn at 12.5 mg/kg group rats were either comparable or better than IZn at 25 mg/kg in terms of zinc retention, CP digestibility, zinc level in serum, liver, bone, and kidney suggesting its better bioavailability simultaneously also reduced fecal excretion of zinc to the environment. Metallothionein mRNA expression was upregulated in NZn at 25 mg/kg and NZn at 50 mg/kg than IZn at 50 mg/kg. Thus, in WAR, NZn at half of the ICAR recommendation (25 mg/kg DM) is as effective as inorganic zinc at 100% of recommended dose.

Sodium ions activated phosphofructokinase leading to enhanced D-lactic acid production by Sporolactobacillus inulinus using sodium hydroxide as a neutralizing agent.

Sporolactobacillus inulinus is a superior D-lactic acid-producing bacterium and proposed species for industrial production. The major pathway for D-lactic acid biosynthesis, glycolysis, is mainly regulated via the two irreversible steps catalyzed by the allosteric enzymes, phosphofructokinase (PFK) and pyruvate kinase. The activity level of PFK was significantly consistent with the cell growth and D-lactic acid production, indicating its vital role in control and regulation of glycolysis. In this study, the ATP-dependent PFK from S. inulinus was expressed in Escherichia coli and purified to homogeneity. The PFK was allosterically activated by both GDP and ADP and inhibited by phosphoenolpyruvate; the addition of activators could partly relieve the inhibition by phosphoenolpyruvate. Furthermore, monovalent cations could enhance the activity, and Na+ was the most efficient one. Considering this kind activation, NaOH was investigated as the neutralizer instead of the traditional neutralizer CaCO3. In the early growth stage, the significant accelerated glucose consumption was achieved in the NaOH case probably for the enhanced activity of Na+-activated PFK. Using NaOH as the neutralizer at pH 6.5, the fermentation time was greatly shortened about 22 h; simultaneously, the glucose consumption rate and the D-lactic acid productivity were increased by 34 and 17%, respectively. This probably contributed to the increased pH and Na+-promoted activity of PFK. Thus, fermentations by S. inulinus using the NaOH neutralizer provide a green and highly efficient D-lactic acid production with easy subsequent purification.

High efficiency cell-recycle continuous sodium gluconate production by Aspergillus niger using on-line physiological parameters association analysis to regulate feed rate rationally.

In this paper, a system of cell-recycle continuous fermentation for sodium gluconate (SG) production by Aspergillus niger (A. niger) was established. Based on initial continuous fermentation result (100.0h) with constant feed rate, an automatic feedback strategy to regulate feed rate using on-line physiological parameters (OUR and DO) was proposed and applied successfully for the first time in the improved continuous fermentation (240.5h). Due to less auxiliary time, highest SG production rate (31.05±0.29gL(-1)h(-1)) and highest yield (0.984±0.067molmol(-1)), overall SG production capacity (975.8±5.8gh(-1)) in 50-L fermentor of improved continuous fermentation increased more than 300.0% compared to that of batch fermentation. Improvement of mass transfer and dispersed mycelia morphology were the two major reasons responsible for the high SG production rate. This system had been successfully applied to industrial fermentation and SG production was greatly improved.

Hydrodynamic cavitation-assisted alkaline pretreatment as a new approach for sugarcane bagasse biorefineries.

Hydrodynamic cavitation (HC) was employed in order to improve the efficiency of alkaline pretreatment of sugarcane bagasse (SCB). Response surface methodology (RSM) was used to optimize pretreatment parameters: NaOH concentration (0.1-0.5M), solid/liquid ratio (S/L, 3-10%) and HC time (15-45min), in terms of glucan content, lignin removal and enzymatic digestibility. Under an optimal HC condition (0.48M of NaOH, 4.27% of S/L ratio and 44.48min), 52.1% of glucan content, 60.4% of lignin removal and 97.2% of enzymatic digestibility were achieved. Moreover, enzymatic hydrolysis of the pretreated SCB resulted in a yield 82% and 30% higher than the untreated and alkaline-treated controls, respectively. HC was found to be a potent and promising approach to pretreat lignocellulosic biomass.

Bioethanol production from sodium hydroxide/hydrogen peroxide-pretreated water hyacinth via simultaneous saccharification and fermentation with a newly isolated thermotolerant Kluyveromyces marxianu strain.

In this study, bioethanol production from NaOH/H2O2-pretreated water hyacinth was investigated. Pretreatment of water hyacinth with 1.5% (v/v) H2O2 and 3% (w/v) NaOH at 25 °C increased the production of reducing sugars (223.53 mg/g dry) and decreased the cellulose crystallinity (12.18%), compared with 48.67 mg/g dry and 22.80% in the untreated sample, respectively. The newly isolated Kluyveromyces marxianu K213 showed greater ethanol production from glucose (0.43 g/g glucose) at 45 °C than did the control Saccharomyces cerevisiae angel yeast. The maximum ethanol concentration (7.34 g/L) achieved with K. marxianu K213 by simultaneous saccharification and fermentation (SSF) from pretreated water hyacinth at 42 °C was 1.78-fold greater than that produced by angel yeast S. cerevisiae at 30 °C. The present work demonstrates that bioethanol production achieved via SSF of NaOH/H2O2-pretreated water hyacinth with K. marxianu K213 is a promising strategy to utilize water hyacinth biomass.

Lithium recovery from brine using a λ-MnO2/activated carbon hybrid supercapacitor system.

Lithium is one of the most important elements in various fields including energy storage, medicine manufacturing and the glass industry, and demands for lithium are constantly increasing these days. The lime soda evaporation process using brine lake water is the major extraction method for lithium, but this process is not only inefficient and time-consuming but also causes a few environmental problems. Electrochemical recovery processes of lithium ions have been proposed recently, but the better idea for the silver negative electrodes used in these systems is required to reduce its cost or increase long term stability. Here, we report an electrochemical lithium recovery method based on a λ-MnO2/activated carbon hybrid supercapacitor system. In this system, lithium ions and counter anions are effectively captured at each electrode with low energy consumption in a salt solution containing various cationic species or simulated Salar de Atacama brine lake water in Chile. Furthermore, we designed this system as a flow process for practical applications. By experimental analyses, we confirmed that this system has high selectivity and long-term stability, with its performance being retained even after repetitive captures and releases of lithium ions.

Intermolecular interactions and 3D structure in cellulose-NaOH-urea aqueous system.

The dissolution of cellulose in NaOH/urea aqueous solution at low temperature is a key finding in cellulose science and technology. In this paper, (15)N and (23)Na NMR experiments were carried out to clarify the intermolecular interactions in cellulose/NaOH/urea aqueous solution. It was found that there are direct interactions between OH(-) anions and amino groups of urea through hydrogen bonds and no direct interaction between urea and cellulose. Moreover, Na(+) ions can interact with both cellulose and urea in an aqueous system. These interactions lead to the formation of cellulose-NaOH-urea-H2O inclusion complexes (ICs). (23)Na relaxation results confirmed that the formation of urea-OH(-) clusters can effectively enhance the stability of Na(+) ions that attracted to cellulose chains. Low temperature can enhance the hydrogen bonding interaction between OH(-) ions and urea and improve the binding ability of the NaOH/urea/H2O clusters that attached to cellulose chains. Cryo-TEM observation confirmed the formation of cellulose-NaOH-urea-H2O ICs, which is in extended conformation with mean diameter of about 3.6 nm and mean length of about 300 nm. Possible 3D structure of the ICs was proposed by the M06-2X/6-31+G(d) theoretical calculation, revealing the O3H···O5 intramolecular hydrogen bonds could remain in the ICs. This work clarified the interactions in cellulose/NaOH/urea aqueous solution and the 3D structure of the cellulose chain in dilute cellulose/NaOH/urea aqueous solution.

Identification of differentially expressed genes in flax (Linum usitatissimum L.) under saline-alkaline stress by digital gene expression.

The salinization and alkalization of soil are widespread environmental problems, and alkaline salt stress is more destructive than neutral salt stress. Therefore, understanding the mechanism of plant tolerance to saline-alkaline stress has become a major challenge. However, little attention has been paid to the mechanism of plant alkaline salt tolerance. In this study, gene expression profiling of flax was analyzed under alkaline-salt stress (AS2), neutral salt stress (NSS) and alkaline stress (AS) by digital gene expression. Three-week-old flax seedlings were placed in 25 mM Na2CO3 (pH11.6) (AS2), 50mM NaCl (NSS) and NaOH (pH11.6) (AS) for 18 h. There were 7736, 1566 and 454 differentially expressed genes in AS2, NSS and AS compared to CK, respectively. The GO category gene enrichment analysis revealed that photosynthesis was particularly affected in AS2, carbohydrate metabolism was particularly affected in NSS, and the response to biotic stimulus was particularly affected in AS. We also analyzed the expression pattern of five categories of genes including transcription factors, signaling transduction proteins, phytohormones, reactive oxygen species proteins and transporters under these three stresses. Some key regulatory gene families involved in abiotic stress, such as WRKY, MAPKKK, ABA, PrxR and ion channels, were differentially expressed. Compared with NSS and AS, AS2 triggered more differentially expressed genes and special pathways, indicating that the mechanism of AS2 was more complex than NSS and AS. To the best of our knowledge, this was the first transcriptome analysis of flax in response to saline-alkaline stress. These data indicate that common and diverse features of saline-alkaline stress provide novel insights into the molecular mechanisms of plant saline-alkaline tolerance and offer a number of candidate genes as potential markers of tolerance to saline-alkaline stress.