High Salt Concentrations Research Articles

Repercussions of the Maternal Obesogenic Diet on the Oxidative Balance and Pancreatic Metabolism in Male Juvenile Offspring

Background/Objectives: The consumption of diets with high fat, salt, and sugar content has been associated with increasing the risk of developing a range of pathologies, including cardiovascular disease, obesity, and diabetes. Furthermore, there is growing evidence to suggest a relationship between variation in the nutritional environment and pancreatic dysregulation, which may be a consequence of oxidative stress. This study aimed to examine the effects of a high-fat, high-carbohydrate (obesogenic) maternal diet during pregnancy and lactation on the metabolic health and pancreatic structure of rat offspring. Methods: Pregnant rats were divided into two groups: one fed a standard diet and the other an obesogenic diet. After weaning, male pups from both groups were fed the same diet until they were 30 days old, which is when they were euthanized. Results: Metabolic and murinometric changes: Increased body weight and pancreas size, elevated blood glucose and cholesterol levels, and reduced glucose tolerance (which is indicative of the beginning of insulin resistance). Oxidative stress: Higher levels of oxidative damage markers and decreased antioxidants in the pancreas, suggesting a state of oxidative stress in this organ. Changes in pancreatic structure: Increased size and number of pancreatic islets and decreased size and number of pancreatic acini. Conclusions: A maternal obesogenic diet induces metabolic alterations, increases oxidative stress, and causes changes in the structure of the pancreas in rat offspring, suggesting a higher risk of developing metabolic diseases such as type 2 diabetes in adulthood.

Electrolyte Strategies at Extreme Temperatures for Aqueous Zinc Batteries

The state of aqueous zinc batteries at extreme temperature environment is an important parameter for their widespread application. However, low ionic conductivity and sluggish ionic diffusion at low temperature, and aggravated untoward reactions in the interface of electrode‐electrolyte at high temperature, seriously limit the practical application of zinc‐ion batteries. Currently, numerous zinc‐ion batteries capable of operating within a broad temperature range have been proposed. Herein, the reason of the performance decline at extreme temperature is discussed from the perspective of thermodynamics and dynamics. Then, from the additives/co‐solvents, high concentration salts, hydrogels and other aspects, the main strategies of electrolyte are introduced in detail. Finally, the possible directions to further improve the high and low temperature performance of zinc‐ion batteries are proposed. It is hoped that this review will be helpful to the design and manufacture of wide temperature zinc‐ion battery electrolyte and provide reference for the application of rechargeable zinc‐ion battery in extreme environment.

Nitrogen removal from multi-electrolyte saline wastewater via mainstream anammox in warm climate conditions.

High salts concentrations in wastewater hinder its biological treatment. Recent research has investigated the inhibitory effect of salinity on the anammox process, mainly focusing on NaCl. Thus, the inhibition caused by multi-electrolytes salinity on freshwater anammox bacteria remains unclear. In this study, the anammox process was evaluated for the treatment of multi-electrolyte saline wastewater (NaCl, MgCl2, and CaCl2) during 684 days in three operational phases. In Phase 1, the anammox inoculum was successfully adapted from sidestream (232 mgN.L-1) to mainstream (60 mgN.L-1) conditions, with no damage to the reactor performance, at an hydraulic retention time of 1.4h. In Phase 2, salinity was gradually increased in the synthetic medium to adapt the freshwater anammox bacteria. The anammox bacteria tolerated a total salinity of 0.72wt% (in g.L-1: 4.7 NaCl, 2.0 MgCl2, and 0.6 CaCl2), achieving an 84.3±0.8% nitrogen removal efficiency. The presence of salts favored the Ca. Jettenia genus over Ca. Brocadia after long-term exposure to salinity. Finally, in Phase 3, anaerobically pre-treated saline wastewater (0.72wt%) was applied to the anammox reactor. The presence of residual organic matter (53 mgCOD.L-1; COD/N of 0.86) resulted in partial deviation of the metabolic pathway from anammox to, especially, nitrite heterotrophic denitrification, resulting in the accumulation of ammonia-N in the effluent. Even so, the anammox process was predominant, being responsible for 83% of the nitrogen removal. The presence of both organic matter and salinity led to a shift in dominance from the Ca. Jettenia genus to Ca. Brocadia.

Wheat β-glucan reduces obesity and hyperlipidemia in mice with high-fat and high-salt diet by regulating intestinal flora.

Unbalanced diets, characterized by high fat or high salt content, are contributing to the obesity epidemic. Wheat bran, recognized as a promising by-product, has the potential to regulate metabolic disorders (MD) associated with obesity. Beta-glucan (BG) has multiple biological activities, but the effect of BG in wheat bran on MD remains unclear. Therefore, this study aimed to investigate the effects of wheat BG (WBG) on dyslipidemia and gut microbiota dysregulation in high fat (HF) or high fat-high salt (HFHS) fed mice. The results demonstrated that WBG significantly reduced the weight gain of mice fed with HF and HFHS diets (from 9.74g to 2.43g and from 6.74g to 2.48g, respectively). Additionally, WBG led to significant reductions in TG (26.26% in HFG and 33.78% in HFHSG) and TC (34.69% in HFG) levels. The liver and adipocyte damage were also reduced after dietary supplementation with WBG. Moreover, WBG significantly reduced Firmicutes/Bacteroidetes ratio (9.52 at HF, 0.62 at HFG, 17.38 at HFHS and 0.61 at HFHSG). Concurrently, there was a reduction in acetic acid levels observed at rates of 26.11% for HF and 32.18% for HFHS. Additionally, WBG reduced the abundance of Coriobacteriaceae UCG-002, Romboutsia, Faecalibaculum, and Enterorhabdus that positively associated with obesity. These changes in gut microbiota may explain the anti-obesity and anti-hyperlipidemia effects of WBG. In conclusion, our findings suggest that WBG is a promising dietary supplement. Our research can provide new insights into the development of foods rich in dietary fiber.

N-terminal domain swapping: A new paradigm for spermidine/spermine N-acetyltransferase (SSAT) protein structures?

Enterococcus faecalis is a multi-drug-resistant human pathogen that is found in a variety of environments and is challenging to treat. Under stress conditions, some bacteria regulate intracellular polyamine concentrations via polyamine acetyltransferases to reduce their toxicity. The E. faecalis genome encodes two polyamine acetyltransferases: PmvE and BltD. Both of these proteins belong to the Gcn5-related N-acetyltransferase (GNAT) superfamily. It is unclear why there are two enzymes with similar substrate specificities in this organism. To better understand the structure/function relationship of the E. faecalis BltD enzyme, we determined its crystal structure and performed additional assays to explore its oligomeric state and enzymatic activity. The goal was to determine whether there were structural or catalytic differences between this enzyme and other polyamine acetyltransferases that could explain this redundancy and be exploited for future development of targeted inhibitors for this important human pathogen. We found the BltD enzyme was structurally unique due to its N-terminal domain swapped dimer. However, this enzyme adopts a catalytically active monomer rather than dimer in solution. This indicates the crystal structure we obtained may represent a state that forms at high protein and salt concentrations and at low pH used during crystallization. The BltD dimer found in the crystal may represent a unique view of how an inhibitory peptide or molecule could be designed to occupy its active site. Additionally, this structure shows the extensive flexibility of the N-terminal portion of the E. faecalis BltD enzyme.

Hydrophobic-unit-regulated hydrogel electrolytes with high water content and low salt concentration for high-voltage aqueous batteries

Hydrophobic-unit-regulated hydrogel electrolytes with high water content and low salt concentration for high-voltage aqueous batteries

The Potential Role of Brassica napus Metallothioneins in Salt Stress and Interactions with Plant Growth-Promoting Bacteria

Background/Objectives: Plant metallothioneins (MTs) are low-molecular-weight proteins involved in heavy metal binding and response to stress conditions. This work aimed to analyse canola (Brassica napus L.) MTs (BnMT1-4) response to salinity and plant interaction with bacteria. Methods: (1) We tested germination and canola growth and development in the presence of sodium chloride and bacteria Serratia plymuthica; (2) We analysed phytohormones content using LC-MS/MS; (3) We identified in silico cis-regulatory elements in promoters of BnMT1-4 genes; and (4) we investigated BnMT1-4 genes’ expression in B. napus. Results: Under saline conditions, canola germination and plant growth were notably inhibited, whereas inoculation of seeds with S. plymuthica significantly stimulated the analysed physiological traits of B. napus. The content of auxin, abscisic acid, jasmonates, gibberellins, and salicylic acid in B. napus was significantly affected by salinity and modulated by S. plymuthica presence. The promoter regions of the BnMT1-4 genes contain numerous regulatory elements controlled by light, hormones, and various stresses. Interestingly, the expression of BnMT1-3 genes was down-regulated under salt stress, while BnMT4 transcript levels increased strongly at the highest salt concentrations with and without S. plymuthica present. Conclusions: The results show that BnMT genes are differently affected by salinity and bacteria S. plymuthica and significantly correlate with particular phytohormones content in canola tissues, confirming the diversified functions of MTs in plant responses to changing environment.

Halobellus marinus sp. nov., Halobellus ordinarius sp. nov., Halobaculum rarum sp. nov., and Halorarum halobium sp. nov., halophilic archaea isolated from marine solar salt and a saline lake.

Four halophilic archaeal strains were isolated from sea salt and a saline lake in China. Based on phylogenetic and phylogenomic analyses, the four strains are related to the genera of Halobellus, Halobaculum, and Halorarum within the family Haloferacaceae. The four strains possess genes responsible for carotenoid synthesis, maintenance of a high internal salt concentration, as well as diverse enzymes with biotechnological potential. The average nucleotide identity (ANI), average amino acid identity (AAI), and digitalDNA-DNA hybridization (dDDH) values among these four strains and their related species were lower than the established thresholds proposed for species demarcation. Strains DFY28T, ZY16T, QDC11T, and XH14T were distinguished from related species based on a variety of phenotypic characteristics. The major polar lipids of these four strains were similar to those of respective relatives within the genera Halobellus, Halobaculum, and Halorarum. The phenotypic, phylogenetic, and genome-based analyses suggest that strains DFY28T (= CGMCC 1.17470T = JCM 34310T), ZY16T (= CGMCC 1.17476T = JCM 34311T), QDC11T (= MCCC 4K00127T = KCTC 4308T), and XH14T (= CGMCC 1.17028T = JCM 34145T) represent four novel species of the genera Halobellus, Halobaculum and Halorarum, for which the names Halobellus marinus sp. nov., Halobellus ordinarius sp. nov., Halobaculum rarum sp. nov., and Halorarum halobium sp. nov. are proposed.

Constructing Anion Solvation Microenvironment Toward Durable High-Voltage Sodium-Based Batteries.

Sodium-based rechargeable batteries are some of the most promising candidates for electric energy storage with abundant sodium reserves, particularly, sodium-based dual-ion batteries (SDIBs) perform advantages in high work voltage (≈5.0V), high-power density, and potentially low cost. However, irreversible electrolyte decomposition and co-intercalation of solvent molecules at the electrode interface under a high charge state are blocking their development. Herein, a high-salt concentration microenvironment is created and proposed by tailoring the solvation structures of charge carriers including both cations and anions, which maintains highly oxidation-resistant contact ion pairs and ion aggregates and provides a high ion conductivity. The tailored solvation structure makes a great contribution to protecting the graphite cathode from electrolyte oxidation, solvent co-intercalation, and structural degradation by constructing a robust cathode-electrolyte interphase with standout electrochemical stability. Based on this, the SDIBs achieved an excellent high-voltage cycling stability with 81% capacity retention after 10000 cycles and the battery showed an improved rate performance with 97.4 mAh g-1 maintained at 100C. It is identified that regulating anion solvation structure is responsible for the stable interface chemistry and enhanced reaction kinetics, which provides deep insight into the compatibility design between the electrolyte and specialized charge storage in electrodes.

СОСТОЯНИЕ И ПЕРСПЕКТИВЫ РАЗВИТИЯ ПРОИЗВОДСТВА ПРОДУКЦИИ ИЗ ПАПОРОТНИКА

The purpose of research is to study the possibility of using salted fern to expand its use for food production. Objectives: to analyze the data presented in the Internet resources about manufacturers of products using the Orlyak fern in Russia; to study the structure of the range of products based on fern; to formulate recommendations on the processing and use of salted fern for the development of new types of food products. 66 companies engaged in the processing of fresh ferns with the production of salted, dried ferns, as well as manufacturers of semi-finished products, culinary products using salted ferns, have been identified. Producers of fern products are concentrated in the Siberian Federal District (SFD), their share is 65 %, 3 times less falls on the Far Eastern Federal District of the FEFD – 23 %. Among the subjects of the federation included in the Siberian Federal District, the Novosibirsk Region (23 %), the Republic of Khakassia and the Krasnoyarsk Region (21 % each), the Irkutsk, Kemerovo, Omsk Regions and Altai Region in total account for 35 %. An analysis of the structure of the product range revealed that in Russia, 57 % of the total volume of processed fern is salted semi-finished product with a high content of table salt, further use of salted fern after soaking in water in the preparation of cold dishes and snacks. A small share (16 %) is occupied by various semi-finished products: stuffed pancakes, meat zrazy and dumplings. Based on the study, recommendations are given to expand the possibilities of using fern, which meet the needs of world trends and consumers.

Dairy consumption has a partial inverse association with systolic blood pressure and hypertension in populations with high salt and low dairy diets: cross-sectional data analysis from the Iwaki Health Promotion Project.

The prevalence of hypertension in Japan remains high, owing to the high salt content of the typical Japanese diet. Dairy-based foods may reduce blood pressure and hypertension risk. However, dairy consumption is low in Japan, and the relationships between dairy intake and blood pressure or the mechanisms by which dairy products affect blood pressure are not fully understood. This cross-sectional study was conducted as part of the Iwaki Health Promotion Project in Aomori, Japan. A total of 1071 participants were included from those who underwent annual medical checkups in June 2015. Adjusted multivariate linear and logistic regression analyses were performed to analyze the relationships between dairy consumption and blood pressure or hypertension risk. Comprehensive blood biomarker measurements were also performed. Whole- and high-fat dairy consumption was found to have significant inverse associations with systolic blood pressure (SBP) for all participants (β = -0.0213, P = 0.044) and with SBP and systolic hypertension risk for non-users of antihypertensive medicines (β = -0.0306, P = 0.011; and OR = 0.9927, P = 0.016; respectively). Three blood biomarkers related to phosphorus metabolism (inorganic phosphorus, intact parathyroid hormone, and interleukin-6) were associated with both dairy consumption and SBP. Dairy consumption had a partial inverse association with SBP and hypertension risk in a Japanese population with high salt and low dairy consumption. Analysis of blood biomarkers suggested that phosphorus metabolism is involved in the associations between dairy consumption and blood pressure.

Straw return was more beneficial to improving saline soil quality and crop productivity than biochar in the short term.

Salinized soil often exhibits high salt content and low nutrient availability, leading to the reduction of soil ecosystem function and crop productivity. Although straw return has profound effects on saline soil improvement, how soil quality index (SQI), soil ecosystem multifunctionality (EMF), and crop yield respond to different organic ameliorants remain unclear. Herein, a field experiment was established to explore the influence of various straw management strategies (no organic ameliorant, CK; corn straw return, CS; and corn straw biochar return; CB) on the saline soil functions and crop productivity. In relation to CK and CB, CS significantly improved SQI by 52% and 35%, respectively. This may be due to the decreased soil salt (especially soluble Na+) and increased available nutrients under corn straw return. Furthermore, CS increased soil EMF than CK by 71% and CB by 39%, which was caused by the increased activities of 1,4-β-glucosidase, β-1,4-N-acetyl-glucosaminidase, and leucine aminopeptidase. The linear model further supported that soil enzyme activities are positively related to available nutrient contents and negatively correlated with salt content. Moreover, the crop yield under CS significantly increased by 22% compared to CK. Also, soil quality positively influenced crop yield, with soil salt and available phosphorus being the primary influencing factors. However, crop yield was not sensitive to soil EMF. In summary, straw return was more beneficial to improving soil quality and crop productivity than biochar in the short term in saline soils.

Creating a Halotolerant Degrader for Efficient Mineralization of p-Nitrophenol-Substituted Organophosphorus Pesticides in High-Saline Wastewater.

The bioaugmentation performance is severely reduced in the treatment of high-saline pesticide wastewater because the growth and degradation activity of pesticide degraders are significantly inhibited by high salt concentrations. In this study, a heterologous biodegradation pathway comprising the seven genes mpd/pnpABCDEF responsible for the bioconversion of p-nitrophenol (PNP)-substituted organophosphorus pesticides (OPs) into β-oxoadipate and the genes encoding Vitreoscilla hemoglobin (VHb) and green fluorescent protein (GFP) were integrated into the genome of a salt-tolerant chassis Halomonas cupida J9, to generate a genetically engineered halotolerant degrader J9U-MP. RT-PCR assays demonstrated that the nine exogenous genes are successfully transcribed to mRNA in J9U-MP. Gas chromatography analysis of methyl parathion (MP) and its intermediates demonstrated that the expressed MP hydrolase and PNP-degrading enzymes PnpABCD show obvious degradation activity toward the specific substrates in J9U-MP. Stable isotope analysis showed that J9U-MP is able to efficiently convert 13C6-PNP into 13CO2, demonstrating the complete mineralization of MP in high-salt media. J9U-MP is genetically stable during passage culture, and genomic integration of exogenous genes does not negatively influence the growth of J9U-MP. Under oxygen-limited conditions, VHb-expressing J9U-MP does not show obvious growth inhibition and a significant reduction in the MP degradation rate. A real-time monitoring system with enhanced GFP is used to track the motion and activity of J9U-MP during bioremediation. Moreover, 50 mg/L MP and its intermediates (i.e., PNP and HQ) were completely degraded by J9U-MP within 12 h in wastewater supplemented with 60 g/L NaCl. After 3 days of incubation, 25 mg/L 13C6-PNP was converted into 13CO2 by J9U-MP in wastewater supplemented with 60 g/L NaCl. Our results highlight the power of synthetic biology for creating new halotolerant pollutant-mineralizing strains. The strong competitive advantages of J9U-MP in high-salinity and low-oxygen environments make this degrader suitable for in situ bioaugmentation of OP wastewater.

Discovery and characterization of a new β-glucosidase from Wolfiporia cocos through RNA sequencing and heterologous expression in Escherichia coli.

De novo RNA-sequencing of Wolfiporia cocos mycelia cultured with filter paper composed of cellulose as the sole carbon source revealed a total of five expressed β-glucosidase genes. Among these, the β-glucosidase named Wcbg1B-1, which is composed of 539 amino acid residues and belongs to the GH1 family, had the highest mRNA abundance, accounting for 65% of the total mRNA of the five expressed β-glucosidases. The recombinant Wcbg1B-1 was successfully expressed in Escherichia coli, with an optimal pH of 6.0 and an optimal temperature of 40°C using p-nitrophenyl-β-d-glucopyranoside (pNPG) as the substrate. Recombinant Wcbg1B-1 has a Km value of 0.32mmol/L, a Vmax of 416μmol/min/mg and specific activity of 461.5U/mg. Recombinant Wcbg1B-1 has strict β-glycosidic bond specificity, the highest hydrolysis activity to cellobiose, followed by lactose, and the lowest hydrolysis activity to gentian. Recombinant Wcbg1B-1 exhibits hydrolytic activity towards macromolecular cellulose such as sodium carboxymethyl cellulose, microcrystalline cellulose and filter paper. Additionally, its high tolerance to high concentrations of glucose and salt makes it more practical for cellulose hydrolysis. Recombinant Wcbg1B-1 can hydrolyze lactose at low temperatures, indicating that it could also be a potential biocatalyst for lactose-reduced dairy industry.

Insights into the salt levels in bread offers in Slovenia: trends and differences.

Bakery products are considered as one of main dietary sources of sodium/salt in Slovenia. Our main objective was to assess the salt content in bread in Slovenia, focusing into different bread categories and sales channels. The data collected in 2022 was compared with year 2012. A follow-up study on salt content of bread sold in Slovenia was conducted. Bread samples were purchased in large retail shops and smaller bakeries across 11 statistical regions of Slovenia. Sodium content was determined by inductively coupled plasma mass spectrometry; salt content was calculated by multiplying sodium content with 2.54, assuming all sodium corresponds to sodium chloride. In 2022, 178 bread samples were purchased and analyzed. Weighted mean salt content in bread was 1.35 (95% CI 1.28-1.42) g/100 g in 2012, and 1.26 (95% CI 1.22-1.29) g/100 g in 2022, showing a 7% decrease. Notable differences in the salt content were observed between various bread subcategories and retail environments. In addition, a significant difference was observed between white wheat bread sold in large retail shops and smaller bakeries, where a higher salt content was observed. While study results show small decrease in the salt content in bread in Slovenia in last decade, the salt reduction targets set by the WHO have not been met. Additional efforts are needed to stimulate bread reformulation with reducing salt content.

Ion Networks in Water-based Li-ion Battery Electrolytes.

ConspectusWater-in-salt electrolytes (WiSEs) are promising electrolytes for next-generation lithium-ion batteries (LIBs), offering critical advantages like nonflammability and improved safety. These electrolytes have extremely high salt concentrations and exhibit unique solvation structures and transport mechanisms dominated by the formation of ion networks and aggregates. These ion networks are central to the performance of WiSEs, govern the transport properties and stability of the electrolyte, deviating from conventional dilute aqueous or organic electrolytes.The availability of free water molecules is significantly reduced in WiSEs, leading to a shift in the solvation environment. Lithium ions (Li+) typically travel with their solvation shells in dilute solutions and form stronger interactions with anions, resulting in the formation of complex ion aggregates. Despite the high viscosity of WiSEs, they exhibit surprisingly high ionic conductivity attributed to the decoupling of viscosity and ionic mobility. Instead of moving through free water, Li+ ions are transported along the pathways formed by the ion networks, minimizing direct solvent interaction and enhancing mobility.Advanced spectroscopic techniques, such as infrared IR pump-probe (IR-PP) and two-dimensional IR (2D-IR) spectroscopy, and molecular dynamics (MD) simulations have illuminated the critical role of these ion networks in facilitating transport. These studies have shown that even at extreme salt concentrations, some water molecules retain properties similar to bulk water, essential for fast ion movement. In WiSEs, bulk-like water molecules form transient hydrogen-bond networks that serve as conduits for Li+ ions, while anion-bound water molecules play a less active role in transport due to their slower dynamics.As the salt concentration increases, the structure of WiSEs becomes more dominated by 3D ion networks. MD simulations reveal that these networks, stabilized by chaotropic anions such as bis(trifluoromethanesulfonyl)imide (TFSI-), disrupt the hydrogen-bonding network of water and provide a stable, interconnected structure that supports the movement of Li+ ions. The formation of these extensive ion networks is critical for maintaining ionic mobility and the electrochemical stability of the electrolyte.The shift from traditional vehicular transport mechanisms to structural diffusion is a hallmark of WiSEs. Li+ ions no longer move with their solvation shells but hop between coordination sites within the ion network. This structural diffusion mechanism enables high ionic mobility despite the reduced presence of water and the increased viscosity of the solution. In conclusion, the formation of ion networks and aggregates in WiSEs not only stabilizes the electrolyte but also drives an unconventional ion transport mechanism. By understanding and controlling these aggregates, WiSEs offer a pathway toward safer, high-performance electrolytes for LIBs and other aqueous energy storage technologies.

Induction of a viable but nonculturable state in Vibrio parahaemolyticus by a high concentration of salt and its impact on fatty acid composition profile and membrane potential

Induction of a viable but nonculturable state in Vibrio parahaemolyticus by a high concentration of salt and its impact on fatty acid composition profile and membrane potential

Purification of micrococcal nuclease for use in ribosomal profiling of high-salinity extremophiles

Nucleases, that is, enzymes that catalyze the hydrolysis of phosphodiester bonds in nucleic acids, are essential tools in molecular biology and biotechnology. Staphylococcus aureus nuclease is particularly interesting due to its thermostability and Ca2+ dependence, making it the prime choice for applications where nuclease modulation is critical, such as ribosome profiling in bacteria and halophilic archaea. The latter poses a technical and economical challenge: high salt reaction conditions are essential for maintaining ribosome integrity but negatively impact the micrococcal nuclease (MNase) activity, necessitating using large amounts of nuclease to achieve efficient cleavage. Here, we set out to generate an optimized production protocol for two forms of MNase-fully processed MNaseA and the 19 amino acid propeptide-containing MNaseB-and to biochemically benchmark them against a commercial nuclease. Our results show that both MNases are highly active in normal reaction conditions, but MNaseA maintains higher enzymatic activity in high salt concentrations than MNaseB. MNaseA also retains >90% of its activity after multiple freeze-thaw cycles when stored at-80 °C in a buffer containing 5% glycerol. Importantly, ribosome profiling experiments in the haloarchaeon Haloferax volcanii demonstrated that MNaseA produces ribosome footprints and hallmarks of active translation highly comparable to those obtained with the commercial nuclease, making it a suitable alternative for high-salt ribosome profiling applications. In conclusion, our method can be easily implemented for efficient MNaseA production, thereby providing access to an effective, robust, and cost-efficient alternative to commercial nucleases, as well as facilitating future translation studies into halophilic organisms.

Efficient capture of 99TcO4-/ReO4- via node and linker bifunctional anion exchange covalent organic frameworks.

In nuclear wastewater treatment, ion-scavenging materials designed to trap 99TcO4- is urgently needed. However, strong acid/base, high radiation and high salt concentration of nuclear wastewater usually result in inadequate stability and adsorption capacity of the adsorbent. Herein, we report a new class of bifunctional anion-exchange olefin-linked COF (BPDC-MTMP) prepared via Knoevenagel condensation reactions, the first example exploring the synergistic integration of positively charged fragments at both nodes and linkers. Surprisingly, BPDC-MTMP exhibits a record ReO4- (a non-radioactive surrogate of 99TcO4-) adsorption capacity up to 1593.21 mg g-1, its outstanding adsorption capacity can be attributed to the synergistic enhancement of the positively charged fragments of the nodes and linkers leading to a significant increase in the positive charge density and the number of anion exchange sites. BPDC-MTMP's hydrophobicity is enhanced by the highly conjugated bulky alkyl skeleton, the affinity toward ReO4- and chemical stability are therefore significantly improved, ReO4- can be selectively and reversibly extracted even under strong acid/base and high salt concentration solutions. This study illustrates that the node and linker bifunctional anion exchange COF is of great potential for ReO4-/99TcO4- trapping, which provides a new method to design high-performance adsorbents for the treatment of nuclear wastewater.

Mixed-mode separation of antisense oligonucleotides using a single column with complementary anion-exchange and hydrophobic interaction chromatography approaches

The current study investigates the use of mixed-mode chromatography as a combination of anion-exchange (AEX) and hydrophobic interaction chromatography (HIC) for the analysis and purification of single-stranded antisense oligonucleotides with stereo-controlled phosphorothioate inter- nucleotide linkages.Initially a Scherzo-SS-C18 trimodal stationary phase with reversed-phase/AEX/ cation-exchange (CEX) functionalities is systematically evaluated to reveal the presence of U-shaped retention composed of two retention modes namely AEX and HIC, where the latter was also observed on related trimodal Scherzo SM and SW analogues. For the first time, retention and separation of deprotected oligonucleotides was described on a single mixed-mode column using a combination of AEX and HIC. This methodology was later applied to an alternative reversed-phase/AEX support, Newcrom BH, displaying similar retention trends under dual salt / organic modifier gradients. The merit of the method was assessed on the basis of separations between a phosphodiester (PO) impurity and phosphorothioate (PS) target for an assortment of selected 2’-O-methoxyethyl 18- to 20-mer single-stranded antisense oligonucleotides. Various parameters were evaluated mostly under HIC conditions including organic modifier percentage, type of salt, temperature, pH and type of buffer in the mobile phase. Retention of the oligonucleotides was significantly affected by the acetonitrile composition and the type of salt ((NH4)2SO4, NaBr, NaCl) where the latter NaCl also afforded resolution between the PS target and closely eluting PO impurities. Small changes in pH between 6.5 and 7 using MES and TRIS respectively demonstrated notable differences in retention and resolution. The optimized methods were compared against a range of traditional supports and applied to various mixed-mode analogues, possessing embedded amino acid, complex forming weak cation-exchange and terminal strong cation-exchange functionalities. The mixed-mode supports displayed HIC retention and better resolution between PS target and PO impurity was evident with a more focused 5% acetonitrile gradient over its 10% counterpart. Overall, throughout the study AEX and HIC demonstrated comparable resolution trends.Finally, the optimized HIC method was applied to a selected 18-mer antisense oligonucleotide at semi-preparative scale using the Newcrom BH column, affording purities of 60-80% and recoveries of 52-76%. Although HIC does not provide better separations between PS and PO than AEX, it opens possibilities to operate under non-denaturing conditions and allows purification of samples containing high salt content, as a standalone method or post AEX without a prior desalting step, which can result in 20-30% sample loss.