Regulating Effect of Exogenous α-Ketoglutarate on Ammonium Assimilation in Poplar.

Extensive industrial activities and anthropogenic agricultural practices have led to substantial ammonia release to the environment. Although croplands can act as ammonia sinks, reduced crop production under high concentrations of ammonium has been documented. Alpha-ketoglutarate (AKG) is a critical carbon source, displaying pleiotropic physiological functions. The objective of the present study is to disclose the potential of AKG to enhance ammonium assimilation in poplars. It showed that AKG application substantially boosted the height, biomass, and photosynthesis activity of poplars exposed to excessive ammonium. AKG also enhanced the activities of key enzymes involved in nitrogen assimilation: glutamine synthetase (GS) and glutamate synthase (GOGAT), elevating the content of amino acids, sucrose, and the tricarboxylic acid cycle (TCA) metabolites. Furthermore, AKG positively modulated key genes tied to glucose metabolism and ATP synthesis, while suppressing ATP-depleting genes. Correspondingly, both H+-ATPase activity and ATP content increased. These findings demonstrate that exogenously applying AKG improves poplar growth under a high level of ammonium treatment. AKG might function through sufficient carbon investment, which enhances the carbon-nitrogen balance and energy stability in poplars, promoting ammonium assimilation at high doses of ammonium. Our study provides novel insight into AKG's role in improving poplar growth in response to excess ammonia exposure.

Newly discovered clouting interplay between matrix metalloproteinases structures and novel quaternary Ammonium K21: computational and in-vivo testing.

AIMS AND OBJECTIVES: To analyze anti-MMP mode of action of Quaternary Ammonium Silane (QAS, codenamed as k21) by binding onto specific MMP site using computational molecular simulation and Anti-Sortase A (SrtA) mode of action by binding onto specific site using computational molecular simulation. MATERIALS AND METHODS: In silico Molecular Dynamics (MD) was used to determine the interactions of K21 inside the pocket of the targeted protein (crystal structure of fibroblast collagenase-1 complexed to a diphenyl-ether sulphone based hydroxamic acid; PDB ID: 966C; Crystal structure of MMP-2 active site mutant in complex with APP-derived decapeptide inhibitor. MD simulations were accomplished with the Desmond package in Schrödinger Drug Discovery Suite. Blood samples (~ 0.5 mL) collected into K2EDTA were immediately transferred for further processing using the Litron MicroFlow® PLUS micronucleus analysis kit for mouse blood according to the manufacturer's instructions. Bacterial Reverse Mutation Test of K21 Molecule was performed to evaluate K21 and any possible metabolites for their potential to induce point mutations in amino acid-requiring strains of Escherichia coli (E. coli) (WP2 uvrA (tryptophan-deficient)). RESULTS: Molecular Simulation depicted that K21 has a specific pocket binding on various MMPs and SrtA surfaces producing a classical clouting effect. K21 did not induce micronuclei, which are the result of chromosomal damage or damage to the mitotic apparatus, in the peripheral blood reticulocytes of male and female CD-1 mice when administered by oral gavage up to the maximum recommended dose of 2000 mg/kg. The test item, K21, was not mutagenic to Salmonella typhimurium (S. typhimurium) strains TA98, TA100, TA1535 and TA1537 and E. coli strain WP2 uvrA in the absence and presence of metabolic activation when tested up to the limit of cytotoxicity or solubility under the conditions of the test. CONCLUSION: K21 could serve as a potent protease inhibitor maintaining the physical and biochemical properties of dental structures.

Ammonium treatment inhibits cell cycle activity and induces nuclei endopolyploidization in Arabidopsis thaliana.

MAIN CONCLUSION: This study determined the effect of ammonium supply on the cell division process and showed that ammonium-dependent elevated reactive oxygen species production could mediate the downregulation of the cell cycle-related gene expression. Plants grown under high-ammonium conditions show stunted growth and other toxicity symptoms, including oxidative stress. However, how ammonium regulates the development of plants remains unknown. Growth is defined as an increase in cell volume or proliferation. In the present study, ammonium-related changes in cell cycle activity were analyzed in seedlings, apical buds, and young leaves of Arabidopsis thaliana plants. In all experimental ammonium treatments, the genes responsible for regulating cell cycle progression, such as cyclin-dependent kinases and cyclins, were downregulated in the studied tissues. Thus, ammonium nutrition could be considered to reduce cell proliferation; however, the cause of this phenomenon may be secondary. Reactive oxygen species (ROS), which are produced in large amounts in response to ammonium nutrition, can act as intermediates in this process. Indeed, high ROS levels resulting from H2O2 treatment or reduced ROS production in rbohc mutants, similar to ammonium-triggered ROS, correlated with altered cell cycle-related gene expression. It can be concluded that the characteristic ammonium growth suppression may be executed by enhanced ROS metabolism to inhibit cell cycle activity. This study provides a base for future research in determining the mechanism behind ammonium-induced dwarfism in plants, and strategies to mitigate such stress.

Anti Gram-Positive Bacteria Activity of Synthetic Quaternary Ammonium Lipid and Its Precursor Phosphonium Salt.

Organic ammonium and phosphonium salts exert excellent antimicrobial effects by interacting lethally with bacterial membranes. Particularly, quaternary ammonium lipids have demonstrated efficiency both as gene vectors and antibacterial agents. Here, aiming at finding new antibacterial devices belonging to both classes, we prepared a water-soluble quaternary ammonium lipid (6) and a phosphonium salt (1) by designing a synthetic path where 1 would be an intermediate to achieve 6. All synthesized compounds were characterized by Fourier-transform infrared spectroscopy and Nuclear Magnetic Resonance. Additionally, potentiometric titrations of NH3+ groups 1 and 6 were performed to further confirm their structure by determining their experimental molecular weight. The antibacterial activities of 1 and 6 were assessed first against a selection of multi-drug-resistant clinical isolates of both Gram-positive and Gram-negative species, observing remarkable antibacterial activity of both compounds against Gram-positive isolates of Enterococcus and Staphylococcus genus. Further investigations on a wider variety of strains of these species confirmed the remarkable antibacterial effects of 1 and 6 (MICs = 4-16 and 4-64 µg/mL, respectively), while 24 h-time-killing experiments carried out with 1 on different S. aureus isolates evidenced a bacteriostatic behavior. Moreover, both compounds 1 and 6, at the lower MIC concentration, did not show significant cytotoxic effects when exposed to HepG2 human hepatic cell lines, paving the way for their potential clinical application.

Molecular Insights into the Synergistic Effects of Putrescine and Ammonium on Dinoflagellates.

Ammonium and polyamines are essential nitrogen metabolites in all living organisms. Crosstalk between ammonium and polyamines through their metabolic pathways has been demonstrated in plants and animals, while no research has been directed to explore this relationship in algae or to investigate the underlying molecular mechanisms. Previous research demonstrated that high concentrations of ammonium and putrescine were among the active substances in bacteria-derived algicide targeting dinoflagellates, suggesting that the biochemical inter-connection and/or interaction of these nitrogen compounds play an essential role in controlling these ecologically important algal species. In this research, putrescine, ammonium, or a combination of putrescine and ammonium was added to cultures of three dinoflagellate species to explore their effects. The results demonstrated the dose-dependent and species-specific synergistic effects of putrescine and ammonium on these species. To further explore the molecular mechanisms behind the synergistic effects, transcriptome analysis was conducted on dinoflagellate Karlodinium veneficum treated with putrescine or ammonium vs. a combination of putrescine and ammonium. The results suggested that the synergistic effects of putrescine and ammonium disrupted polyamine homeostasis and reduced ammonium tolerance, which may have contributed to the cell death of K. veneficum. There was also transcriptomic evidence of damage to chloroplasts and impaired photosynthesis of K. veneficum. This research illustrates the molecular mechanisms underlying the synergistic effects of the major nitrogen metabolites, ammonium and putrescine, in dinoflagellates and provides direction for future studies on polyamine biology in algal species.

Antibacterial Activity of Boron Compounds Against Biofilm-Forming Pathogens.

This study aimed to evaluate the antibacterial activity of nine boron derivatives against biofilm-forming pathogenic bacteria. The effect of boron derivatives (CMB, calcium metaborate; SMTB, sodium metaborate tetrahydrate; ZB, zinc borate; STFB, sodium tetra fluorine borate; STB, sodium tetraborate; PTFB, potassium tetra fluor borate; APTB, ammonium pentabo-rate tetrahydrate; SPM, sodium perborate monohydrate; Borax, ATFB, ammonium tetra fluorine borate) on bacteria isolated from blood culture was determined by the minimum inhibitory concentration (MIC) method. Then, biofilm formation potentials on microplates, tubes, and Congo red agar were examined. The cytotoxicity of boron derivatives was determined by using WST-1-based methods. The interaction between the biofilm-forming bacteria, fibroblast cells, and boron derivatives was determined with the infection model. We found that the sodium metaborate tetrahydrate molecule was effective against all pathogens. According to the optical density values detected at 630 nm in microplates, meticillin-resistant Staphylococcus aureus was observed to have the most substantial biofilm ability at 0.257 nm. As a result of cytotoxicity studies, it has been determined that a 1 µg/L concentration of boron derivatives is not toxic to fibroblast L929 cells. In cell culture experiments, these boron derivatives have very serious inhibitory activity against biofilm-forming pathogens in a short treatment period, such as 2-4 h. Furthermore, using these molecules on inanimate surfaces affected by biofilms would be appropriate instead of living cells.

Metabolomic response of striped marsh frog (Limnodynastes peronii) tadpoles exposed to the fire retardant Phos-Chek LC95W.

Climate change and other factors have contributed to an increased frequency and intensity of global wildfires in recent years. Ammonium-based fire retardants are widely used to suppress or delay the spread of fire and have generally been regarded as presenting a low risk of acute toxicity to fauna. However, studies have raised concerns about their potential to cause indirect or sub-lethal effects, and toxicity information regarding the potential for such impacts in aquatic species is limited. To address these knowledge gaps, we used an untargeted metabolomics approach to evaluate the sub-lethal physiological and metabolic responses of striped marsh frog (Limnodynastes peronii) tadpoles exposed to a concentration gradient of the ammonium polyphosphate (APP)-based fire retardant Phos-Chek LC95W (PC). Acute exposure (96 h) to PC significantly altered the relative abundance of 14 metabolites in whole tadpoles. The overall metabolic response pattern was consistent with effects reported for ammonia toxicity and also suggestive of energy dysregulation and osmotic stress associated with alterations to physicochemical water quality parameters in the PC treatments. Results suggest that run-off or accidental application of this formulation into waterways can have significant sub-lethal consequences on the biochemical profiles (i.e., the metabolome) of aquatic organisms and may be a concern for frog species that breed and develop in small, often ephemeral, waterbodies. Our study highlights the benefits of integrating untargeted metabolomics with other ecological and toxicological endpoints to provide a more holistic characterisation of the sub-lethal impacts associated with bushfire-fighting chemicals and with environmental contaminants more broadly.

Antagonizing Effects of Chromium Against Iron-Decreased Glucose Uptake by Regulating ROS-Mediated PI3K/Akt/GLUT4 Signaling Pathway in C2C12.

To investigate the effect of chromium and iron on glucose metabolism via the PI3K/Akt/GLUT4 signaling pathway. Skeletal muscle gene microarray data in T2DM (GSE7014) was selected using Gene Expression Omnibus database. Element-gene interaction datasets of chromium and iron were extracted from comparative toxicogenomics database (CTD). Gene ontology (GO)and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using DAVID online tool. Cell viability, insulin-stimulated glucose uptake, intracellular reactive oxygen species (ROS) level, and protein expression level were measured in C2C12 cells. The bioinformatics research indicated that PI3K/Akt signaling pathway participated in the effects of chromium and iron associated with T2DM. Insulin-stimulated glucose uptake level was significantly higher in chromium picolinate (Cr group) and lower in ammonium iron citrate (FA group) than that for the control group (P < 0.05); chromium picolinate + ammonium iron citrate (Cr + FA group) glucose uptake level was higher than that for the FA group (P < 0.05). Intracellular ROS level was significantly higher in the FAC group than that for the control group (P < 0.05), and that for the Cr + FA group was lower than that for the FA group (P < 0.05). p-PI3K/PI3K, p-Akt/Akt, and GLUT4 levels were significantly lower in the FA group than that for the control group (P < 0.05), and the Cr + FA group had higher levels than the FA group (P < 0.05). Chromium might have a protective effect on iron-induced glucose metabolism abnormalities through the ROS-mediated PI3K/Akt/GLUT4 signaling pathway.

Staphylococcus aureus adapts to the immunometabolite itaconic acid by inducing acid and oxidative stress responses including S-bacillithiolations and S-itaconations.

Staphylococcus aureus is a major pathogen, which has to defend against reactive oxygen and electrophilic species encountered during infections. Activated macrophages produce the immunometabolite itaconate as potent electrophile and antimicrobial upon pathogen infection. In this work, we used transcriptomics, metabolomics and shotgun redox proteomics to investigate the specific stress responses, metabolic changes and redox modifications caused by sublethal concentrations of itaconic acid in S. aureus. In the RNA-seq transcriptome, itaconic acid caused the induction of the GlnR, KdpDE, CidR, SigB, GraRS, PerR, CtsR and HrcA regulons and the urease-encoding operon, revealing an acid and oxidative stress response and impaired proteostasis. Neutralization using external urea as ammonium source improved the growth and decreased the expression of the glutamine synthetase-controlling GlnR regulon, indicating that S. aureus experienced ammonium starvation upon itaconic acid stress. In the extracellular metabolome, the amounts of acetate and formate were decreased, while secretion of pyruvate and the neutral product acetoin were strongly enhanced to avoid intracellular acidification. Exposure to itaconic acid affected the amino acid uptake and metabolism as revealed by the strong intracellular accumulation of lysine, threonine, histidine, aspartate, alanine, valine, leucine, isoleucine, cysteine and methionine. In the proteome, itaconic acid caused widespread S-bacillithiolation and S-itaconation of redox-sensitive antioxidant and metabolic enzymes, ribosomal proteins and translation factors in S. aureus, supporting its oxidative and electrophilic mode of action in S. aureus. In phenotype analyses, the catalase KatA, the low molecular weight thiol bacillithiol and the urease provided protection against itaconic acid-induced oxidative and acid stress in S. aureus. Altogether, our results revealed that under physiological infection conditions, such as in the acidic phagolysome, itaconic acid is a highly effective antimicrobial against multi-resistant S. aureus isolates, which acts as weak acid causing an acid, oxidative and electrophilic stress response, leading to S-bacillithiolation and itaconation.

Study on the mechanism of nitrapyrin microcapsule suspension effectively improving nitrification inhibition rate in black soil.

Nitrification inhibitors (NIs) have been widely applied to inhibit nitrification and reduce N2O emissions in agriculture. However, there are still some shortcomings, e.g. short effective periods, large applying amounts, low effectiveness, easy deactivation and different effect. Thus, a nitrapyrin microcapsule suspension (CPCS) was used as a new experimental material to elaborate its effects on nitrogen transformation and microbial response mechanisms in black soil by cultivation experiments with six treatments of no fertilization (CK), urea, urea+ 0.2 % CPES, urea+ 0.1 % CPCS, urea+ 0.2 % CPCS, and urea+ 0.3 % CPCS. The content of ammonium, nitrate nitrogen, functional microbial activity, degradation rate and adsorption characteristics of CPCS in the soil at different incubating times were determine. Compared with the nitrapyrin emulsifiable concentrate (CPEC) treatment, the degradation rate of CPCS decreased by 21.54 %, the half-life increased by 10.2 days, and the adsorption rate of nitrapyrin on black soil decreased more than 6-fold. CPCS effectively inhibited the transformation of ammonium nitrogen to nitrate nitrogen within more than 42 days. CPCS had a negative effect on amoA gene abundance and a positive effect on nrfA gene abundance. The research results provide a basic theoretical support for the application of CPCS on black soil.

Light-Sensitive Open Channel Block of Ionotropic Glutamate Receptors by Quaternary Ammonium Azobenzene Derivatives.

Glutamate ionotropic receptors mediate fast excitation processes in the central nervous system of vertebrates and play an important role in synaptic plasticity, learning, and memory. Here, we describe the action of two azobenene-containing compounds, AAQ (acrylamide-azobenzene-quaternary ammonium) and QAQ (quaternary ammonium-azobenzene-quaternary ammonium), which produced rapid and fully reversible light-dependent inhibition of glutamate ionotropic receptors. The compounds demonstrated voltage-dependent inhibition with only minor voltage-independent allosteric action. Calcium-impermeable AMPA receptors had weaker sensitivity compared to NMDA and calcium-permeable AMPA receptors. We further revealed that the compounds bound to NMDA and calcium-permeable AMPA receptors in different modes. They were able to enter the wide selectivity filter of AMPA receptors, and strong negative voltages caused permeation into the cytoplasm. The narrow selectivity filter of the NMDA receptors did not allow the molecules to bypass them; therefore, QAQ and AAQ bound to the shallow channel site and prevented channel closure by a foot-in-the-door mechanism. Computer simulations employing available AMPA and NMDA receptor structures readily reproduced the experimental findings, allowing for the structure-based design of more potent and selective drugs in the future. Thus, our work creates a framework for the development of light-sensitive blockers of calcium-permeable AMPA receptors, which are desirable tools for neuroscience.

Phosphatidic acid regulates ammonium uptake by interacting with AMMONIUM TRANSPORTER 1;1 in Arabidopsis.

Ammonium (NH4+) is a key inorganic nitrogen source in cellular amino acid biosynthesis. The coupling of transcriptional and posttranslational regulation of AMMONIUM TRANSPORTER (AMT) ensures that NH4+ acquisition by plant roots is properly balanced, which allows for rapid adaptation to a variety of nitrogen conditions. Here, we report that phospholipase D (PLD)-derived phosphatidic acid (PA) interacts with AMT1;1 to mediate NH4+ uptake in Arabidopsis (Arabidopsis thaliana). We examined pldα1 pldδ-knockout mutants and found that a reduced PA level increased seedling growth under nitrogen deficiency and inhibited root growth upon NH4+ stress, which was consistent with the enhanced accumulation of cellular NH4+. PA directly bound to AMT1;1 and inhibited its transport activity. Mutation of AMT1;1 R487 to Gly (R487G) resulted in abolition of PA suppression and, subsequently, enhancement of ammonium transport activity in vitro and in vivo. Observations of AMT1;1-GFP showed suppressed endocytosis under PLD deficiency or by mutation of the PA-binding site in AMT1;1. Endocytosis was rescued by PA in the pldα1 pldδ mutant but not in the mutant AMT1;1R487G-GFP line. Together, these findings demonstrated PA-based shutoff control of plant NH4+ transport and point to a broader paradigm of lipid-transporter function.

Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed with a tempered barley-based diet with canola oil.

A dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ±â€…14.4 kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0 mg/kg dry matter (DM; control), 50 mg/kg DM, 75 mg/kg DM, 100 mg/kg DM, and 125 mg/kg DM. Daily CH4 production was measured on day 7 (last day of starter diet), day 14 (last day of the first intermediate diet), and day 21 (last day of the second intermediate diet) of the adaptation period and on days 28, 49, 70, 91, and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement postfeeding, and prefeeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH, and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, VFA concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.

Livestock methane (CH4) is the main source of greenhouse gases (GHGs) in agriculture, contributing to 11.6% of global GHGs emissions from human-related activities. Therefore, mitigating CH4 emissions from ruminant animals is a great opportunity for meeting the current climate targets. In this experiment, increasing inclusion rates of a promising CH4-mitigating compound, 3-nitrooxypropanol (3-NOP, from 50 to 125 mg of 3-NOP/kg of dry matter [DM]), were added to a barley-based feedlot diet containing 25 ppm of monensin and 7% fat (DM-basis) and fed to Angus steers. Under these conditions, increasing inclusion rate of 3-NOP reduced both production and yield of CH4 by up to 90%. Rumen fermentation, feed intake, and average daily gain were not affected by the 3-NOP dose. Our results on the potential CH4 suppression of 3-NOP may assist the feedlot industry towards sustainability by lowering its GHG output.

Preparation of Gelatin-Quaternary Ammonium Salt Coating on Titanium Surface for Antibacterial/Osteogenic Properties.

Titanium (Ti) and its alloys are widely used in medical treatment, engineering, and other fields because of their excellent properties including biological activity, an elastic modulus similar to that of human bones, and corrosion resistance. However, there are still many defects in the surface properties of Ti in practical applications. For example, the biocompatibility of Ti with bone tissue can be greatly reduced in implants due to a lack of osseointegration as well as antibacterial properties, which may lead to osseointegration failure. To address these problems and to take advantage of the amphoteric polyelectrolyte properties of gelatin, a thin layer of gelatin was prepared by electrostatic self-assembly technology. Diepoxide quaternary ammonium salt (DEQAS) and maleopimaric acid quaternary ammonium salt (MPA-N+) were then synthesized and grafted onto the thin layer. The cell adhesion and migration experiments demonstrated that the coating has excellent biocompatibility, and those grafted with MPA-N+ promoted cell migration. The bacteriostatic experiment showed that the mixed grafting with two ammonium salts had excellent bacteriostatic performance against Escherichia coli and Staphylococcus aureus, with bacteriostasis rates of 98.1 ± 1.0% and 99.2 ± 0.5%, respectively.

Nitric Oxide Induced by Ammonium/Nitrate Ratio Ameliorates Low-Light Stress in Brassica pekinesis: Regulation of Photosynthesis and Root Architecture.

Low-light intensity affects plant growth and development and, finally, causes a decrease in yield and quality. There is a need for improved cropping strategies to solve the problem. We previously demonstrated that moderate ammonium:nitrate ratio (NH4+:NO3-) mitigated the adverse effect caused by low-light stress, although the mechanism behind this alleviation is unclear. The hypothesis that the synthesis of nitric oxide (NO) induced by moderate NH4+:NO3- (10:90) involved in regulating photosynthesis and root architecture of Brassica pekinesis subjected to low-light intensity was proposed. To prove the hypothesis, a number of hydroponic experiments were conducted. The results showed that in plants exposed to low-light intensity, the exogenous donors NO (SNP) and NH4+:NO3- (N, 10:90) treatments significantly increased leaf area, growth range, and root fresh weight compared with nitrate treatment. However, the application of hemoglobin (Hb, NO scavenger), N-nitro-l-arginine methyl ester (L-NAME, NOS inhibitor), and sodium azide (NaN3, NR inhibitor) in N solution remarkably decreased the leaf area, canopy spread, the biomass of shoot and root, the surface area, and volume and tips of the root. The application of N solution and exogenous SNP significantly enhanced Pn (Net photosynthetic rate) and rETR (relative electron transport rates) compared with solo nitrate. While all these effects of N and SNP on photosynthesis, such as Pn, Fv/Fm (maximum quantum yield of PSII), Y(II) (actual photosynthetic efficiency), qP (photochemical quenching), and rETR were reversed when the application of Hb, L-NAME, and NaN3 in N solution. The results also showed that the N and SNP treatments were more conducive to maintaining cell morphology, chloroplast structure, and a higher degree of grana stacking of low-light treated plants. Moreover, the application of N significantly increased the NOS and NR activities, and the NO levels in the leaves and roots of mini Chinese cabbage seedlings treated with N were significantly higher than those in nitrate-treated plants. In conclusion, the results of this study showed that NO synthesis induced by the appropriate ammonia-nitrate ratio (NH4+:NO3- = 10:90) was involved in the regulation of photosynthesis and root structure of Brassica pekinesis under low-light stress, effectively alleviating low-light stress and contributing to the growth of mini Chinese cabbage under low-light stress.

Lactobacillus (L. plantarum & L. rhamnosus) and Saccharomyces (S. cerevisiae): effects on performance, biochemical parameters, ammonium ion in manure, and digestibility of broiler chickens.

Two strains of Lactobacillus combined with Baker's yeast (Saccharomyces cerevisiae) used as probiotics were evaluated to replace antibiotics in poultry flocks by reducing ammonia emissions in manure of broilers without comprising performance or health. One-day-old Cobb 500 broilers (600) were fed starter, grower, and finisher diets as control (CON); probiotic S. cerevisiae, inclusion rate at 4.26 × 106 CFU/kg of feed (SCY); probiotic L. plantarum and L. rhamnosus, inclusion rate at 4.35 × 108 CFU/kg of feed (LPR) for each; and a combination of Lactobacillus plantarum and L. rhamnosus at 4.35 × 108 CFU/kg of feed for each plus Saccharomyces cerevisiae and 4.26 × 106 CFU/kg of feed (SWL). The 4 treatments had 5 replicates (pens), each with 30 broilers. Performance was measured weekly as feed consumption, weight gain, BW, and feed conversion ratio (FCR) over a 6-wk grow-out period. Accompanying biochemical analyses included lipase activity of the pancreas, liver weight, and uric acid (UA) concentration in liver. Albumin, total protein, UA, ammonia, and blood urea nitrogen (BUN) were measured in serum. Ammonium (NH4+) in manure and apparent ileal digestibility from digesta were also measured. Significance was determined at P ≤ 0.05. Results showed that biochemical analyses had no significant treatment effect; however, there were significant temporal changes in performance measures for individual treatments. Feed consumption increased over time for all treatments (P = 2.00 × 10-16). CON had lower weight gain in wk 2 (P = 0.013) compared to all treatment and the lowest BW in wk 5 (P = 0.0008) and wk 6 (P = 0.0124) compared to SWL. Specific probiotic strains, with well-defined inclusion rates, and surrounding environmental analyses of present microbes are needed to ascertain effects of probiotics. Other important areas for investigation include 1) confirmation of probiotics present in the digesta/ceca and how they alter the microbiota within the gastrointestinal (GI) tract and 2) the serum heterophil:lymphocyte ratio to further examine potential immune responses to the probiotics.

How anaerobic sludge microbiome respond to different concentrations of nitrite, nitrate, and ammonium ions: a comparative analysis.

High concentrations of ammonium, nitrite, and nitrate always induce inhibition in anaerobic wastewater treatment. Due to the complexity and vulnerability of the microbial community (especially methanogens) in anaerobic sludge, little is understood about its underlying microbial mechanism under such inhibition. In this study, the shifts of microbial communities in anaerobic sludge under increasing levels of nitrite, nitrate, and ammonium ions were compared. Results show that although half maximal inhibitory concentrations (methanogenesis) were different for nitrite, nitrate, and ammonium ions with EC50 values of 12, 30, and 3000 mg N/L, respectively, bacteria genera Kosmotoga and Brooklawnia dominated in all of the three high-stress inhibitory systems. Network analysis and redundancy analysis (RDA) of the microbial community showed the treatments with nitrate and nitrite ions decreased the modularity of anaerobic microorganisms. RDA showed that specific methanogenic activity was positively related to coenzyme F420 under nitrite inhibition (rp = 0.833, p < 0.05) and closely correlated with viability under nitrate inhibition. Gram-positive and nonmotile Brooklawnia genus showed a negative correlation with physiological characteristics in the ammonia treatments, suggesting its high resistance to ammonia.

Effects of nitrate and ammonium on assimilation of nitric oxide by Heterosigma akashiwo.

The harmful alga Heterosigma akashiwo possesses a hybrid nitrate reductase (NR) enzyme, NR2-2/2HbN, which has the potential to convert NO to nitrate for assimilation into biomass. In previous research, NR transcription in H. akashiwo was induced by nitrate while NR activity was inhibited by ammonium. Here, the capacity of H. akashiwo to use NO in the presence of nitrate and/or ammonium was investigated to understand the regulation of NO assimilation. Continuous cultures of H. akashiwo were acclimated to growth on nitrate, ammonium, or a mixture of both. Aliquots from these cultures were spiked with 15N-labeled NO. The expression of genes involved in nitrogen assimilation was evaluated, as well as nitrate reductase activity and assimilation of 15N-labeled nitrogen into algal biomass. Results showed that NO induced expression and activity of NR, and upregulated expression of GOGAT regardless of the presence of other inorganic nitrogen sources, while GS expression decreased over time. Furthermore, 15NO uptake and assimilation was significantly higher in cultures acclimated for growth on ammonium compared to cultures acclimated for growth on nitrate alone. Assimilation of NO may provide H. akashiwo with a competitive advantage in N-poor environments or areas with elevated NO.

Ammonium effects on oxidative stress, telomere length, and locomotion across life stages of an anuran from habitats with contrasting land-use histories.

Understanding the mechanistic implications behind wildlife responses to global changes is a central topic in eco-evolutionary research. In particular, anthropic pollution is known to impact wild populations across the globe, which may have even stronger consequences for species with complex life cycles. Among vertebrates, amphibians represent a paradigmatic example of metamorphosis, and their characteristics make them highly vulnerable to pollution. Here, we tested for differences in the redox status, telomere length, and locomotor performance across life stages of green frogs (Pelophylax perezi) from agrosystem and natural habitats, both constitutively and in response to an experimental ammonium exposure (10 mg/L). We found that larvae from the agrosystem constitutively showed an enhanced redox status (better antioxidant balance against H2O2, lower lipid peroxidation) but shorter telomeres as compared to larvae from the natural environment. The larval redox response to ammonium was, overall, similar in both habitats. In contrast, after metamorphosis, the redox status of individuals from the natural habitat seemed to cope better with ammonium exposure (denoted by lower lipid peroxidation), and differences between habitats in telomere length were no longer present. Intriguingly, while the swimming performance of larvae did not correlate with individual's physiology, metamorphs with lower glutathione reductase activity and longer telomeres had a better jumping performance. This may suggest that locomotor performance is both traded off with the production of reactive oxygen species and potentiated directly by longer telomeres or indirectly by the mechanisms that buffer their shortening. Overall, our study suggests that contrasting land-use histories can drive divergence in physiological pathways linked to individual health and lifespan. Since this pattern was life-stage dependent, divergent habitat conditions can have contrasting implications across the ontogenetic development of species with complex life cycles.

Development of a quaternary ammonium photoswitchable antagonist of NMDA receptors.

NMDA receptors play critical roles in numerous physiological and pathological processes in CNS that requires development of modulating ligands. In particular, photoswitchable compounds that selectively target NMDA receptors would be particularly useful for analysis of receptor contributions to various processes. Recently, we identified a light-dependent anti-NMDA activity of the azobenzene-containing quaternary ammonium compounds DENAQ (diethylamine-azobenzene-quaternary ammonium) and DMNAQ (dimethylamine-azobenzene-quaternary ammonium). Here, we developed a series of light-sensitive compounds based on the DENAQ structure, and studied their action on glutamate receptors in rat brain neurons using patch-clamp method. We found that the activities of the compounds and the influence of illumination strongly depended on the structural details, as even minor structural modifications greatly altered the activity and sensitivity to illumination. The compound PyrAQ (pyrrolidine-azobenzene-quaternary ammonium) was the most active and produced fast and fully reversible inhibition of NMDA receptors. The IC50 values under ambient and monochromic light conditions were 2 and 14 µM, respectively. The anti-AMPA activity was much weaker. The action of PyrAQ did not depend on NMDA receptor activity, agonist concentration, or membrane voltage, making it a useful tool for photopharmacological studies.