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Objective: The purpose of this study was to determine whether xenon post-conditioning affects mTOR signaling as well as endoplasmic reticulum stress (ERS)-apoptosis pathway in rats with spinal cord ischemia/reperfusion injury., Methods: Fifty male rats were randomized equally into sham-operated group (Sham group), I/R model group (I/R group), I/R model+ xenon post-conditioning group (Xe group), I/R model+rapamycin (a mTOR signaling pathway inhibitor) treatment group (I/R+ Rapa group), and I/R model + xenon post- conditioning with rapamycin treatment group (Xe + Rapa group).. In the latter 4 groups, SCIRI was induced by clamping the abdominal aorta for 85 min followed by reperfusion for 4 h. Rapamycin (or vehicle) was administered by daily intraperitoneal injection (4 mg/kg) for 3 days before SCIRI, and xenon post-conditioning by inhalation of 1∶1 mixture of xenon and oxygen for 1 h at 1 h after initiation of reperfusion; the rats without xenon post-conditioning were given inhalation of nitrogen and oxygen (1∶ 1). After the reperfusion, motor function and histopathologic changes in the rats were examined. Western blotting and real-time PCR were used to detect the protein and mRNA expressions of GRP78, ATF6, IRE1α, PERK, mTOR, p-mTOR, Bax, Bcl-2 and caspase-3 in the spinal cord., Results: The rats showed significantly lowered hind limb motor function following SCIRI ( P < 0.01) with a decreased count of normal neurons, increased mRNA and protein expressions of GRP78, ATF6, IRE1α, PERK, and caspase-3, and elevated p-mTOR/mTOR ratio and Bax/Bcl-2 ratio ( P < 0.01). Xenon post-conditioning significantly decreased the mRNA and protein levels of GRP78, ATF6, IRE1α, PERK and caspase-3 ( P < 0.05 or 0.01) and reduced p-mTOR/mTOR and Bax/Bcl-2 ratios ( P < 0.01) in rats with SCIRI; the mRNA contents and protein levels of GRP78 and ATF6 were significantly decreased in I/R+Rapa group ( P < 0.01). Compared with those in Xe group, the rats in I/R+Rapa group and Xe+Rapa had significantly lowered BBB and Tarlov scores of the hind legs ( P < 0.01), and caspase-3 protein level and Bax/Bcl-2 ratio were significantly lowered in Xe+Rapa group ( P < 0.05 or 0.01)., Conclusion: By inhibiting ERS and neuronal apoptosis, xenon post- conditioning may have protective effects against SCIRI in rats. The mTOR signaling pathway is partially involved in this process.

Background: Traumatic brain injury (TBI)-induced acute lung injury (ALI) is a critical condition, and inflammation and apoptosis play essential roles. Molecular hydrogen (H 2 ) exerts anti-inflammatory and anti-apoptotic effects. Our previous work has shown that 42% H 2 can improve TBI. In the current study, we tested the hypothesis that inhalation of hydrogen (42% H 2 , 21% O 2 , balanced nitrogen) for 1 h per day can improve TBI-induced ALI., Methods: Sprague-Dawley male rats were randomly divided into 3 groups. Except for the sham group (group S), rats were subjected to a fluid percussion injury (FPI) and the H 2 treatment group were given inhaled hydrogen for 1 h per day. We evaluated the lung function, pyroptosis and apoptosis at 24 h, 48 h and 72 h., Results: Compared with group S, the rats in the TBI group (group T) showed obvious pulmonary edema after a TBI. Inhalation of high-concentration hydrogen significantly improved the rats. During this process, rats had some tendency to heal on their own, and H 2 also accelerated the self-healing process. Lung injury scores, oxygenation index and pulmonary edema were consistent. Compared with group S, the pyroptosis-related proteins Caspase-1, apoptosis-associated speck-like protein containing CARD (ASC) and Gasdermin-D (GSDM-D) in the lung tissues of the rats in group T were significantly increased after a TBI. In the H 2 treatment group (group H), these proteins were significantly decreased. The levels of IL-1β and IL-18 were significantly increased after TBI while in group H were significantly decreased. At the same time, cleaved caspase-3 and BCL-2/Bax were also changed after H 2 treatment. These demonstrates the powerful ameliorating effect of H 2 on pyroptosis, apoptosis and systemic inflammation. However, rats also had tendency to heal on their own, and H 2 also accelerated the self-healing process at the same time., Conclusions: H 2 improves TBI-ALI, and the mechanism may be due to the decrease of both pyroptosis and apoptosis and the alleviation of inflammation. These findings provide a reference and evidence for the use of H 2 in TBI-ALI patients in the intensive care unit (ICU)., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Anthropogenic disturbances may decrease as we take measures to control them. However, the patterns and mechanisms of post-disturbance ecosystem succession have rarely been studied. Here we reported that disturbance level determined the importance of stochastic relative to deterministic changes in ecosystem components (plant community composition, soil microbial community composition, and soil physicochemical indices), and thus predefined the pattern of post-disturbance ecosystem succession. We proposed a theoretical framework with five disturbance levels corresponding to distinct succession patterns. We conducted a nitrogen addition experiment in a temperate steppe, monitored these ecosystem components during "disturbance" treatment (2010-2014) and post-treatment "succession" (2014-2018). The disturbance level experienced by each component in each treatment was inferred by fitting the observed succession patterns into the theoretical framework. The mean disturbance level of these components was found to increase quadratically with nitrogen addition rate. This was because increasing nitrogen addition reduced the importance of stochastic relative to deterministic changes in these components, and these changes had a quadratic relationship with disturbance level. Overall, our results suggested that by monitoring the importance of stochastic relative to deterministic changes in an ecosystem, we can estimate disturbance levels and predict succession patterns, as well as propose disturbance-level-dependent strategies for post-disturbance restoration., (© 2021. Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature.)

By means of the formation of SeN, the ABT-Se and NDI-Se were developed to detect and visualize endogenous hypobromous acid (HOBr) in live cells. Specifically, the upregulation of HOBr was monitored by NDI-Se during the administration of an immunotherapeutic agent.

Oats (Avena sativa L.) are a healthy food, being high in dietary fibre (e.g. β-glucans), antioxidants, minerals, and vitamins. Understanding the effect of variety and crop management on nutritional quality is important. The response of four oat varieties to increased nitrogen levels was investigated across multiple locations and years with respect to yield, grain quality and metabolites (assessed via GC- and LC- MS). A novel high-resolution UHPLC-PDA-MS/MS method was developed, providing improved metabolite enrichment, resolution, and identification. The combined phenotyping approach revealed that, amino acid levels were increased by nitrogen supplementation, as were total protein and nitrogen containing lipid levels, whereas health-beneficial avenanthramides were decreased. Although nitrogen addition significantly increased grain yield and β-glucan content, supporting increasing the total nitrogen levels recommended within agricultural guidelines, oat varietal choice as well as negative impacts upon health beneficial secondary metabolites and the environmental burdens associated with nitrogen fertilisation, require further consideration., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Nitrogen (N) and phosphorus (P) are macronutrients essential for crop growth and productivity. In cultivated fields, N and P levels are rarely sufficient, contributing to the gap between realized and potential production. Fertilizer application increases nutrient availability, but is not available to all farmers, nor are current rates of application sustainable or environmentally desirable. Transcriptomic studies of cereal crops have revealed dramatic responses to either low N or low P single stress treatments. In the field, however, levels of both N and P may be suboptimal. The interaction between N and P starvation responses remains to be fully characterized., Results: We characterized growth and root and leaf transcriptomes of young maize plants under nutrient replete, low N, low P or combined low NP conditions. We identified 1555 genes to respond to our nutrient treatments, in one or both tissues. A large group of genes, including many classical P starvation response genes, were regulated antagonistically between low N and P conditions. An additional experiment over a range of N availability indicated that a mild reduction in N levels was sufficient to repress the low P induction of P starvation genes. Although expression of P transporter genes was repressed under low N or low NP, we confirmed earlier reports of P hyper accumulation under N limitation., Conclusions: Transcriptional responses to low N or P were distinct, with few genes responding in a similar way to the two single stress treatments. In combined NP stress, the low N response dominated, and the P starvation response was largely suppressed. A mild reduction in N availability was sufficient to repress the induction of P starvation associated genes. We conclude that activation of the transcriptional response to P starvation in maize is contingent on N availability.

The excessive consumption of nitrogen (N) and phosphorus (P), two vital nutrients for living organisms, is associated with negative environmental and health impacts. While food production contributes to a large amount of N and P loss to the environment, very little N and P is consumed as food. Food habits are affected by multiple regulations, including the dietary restrictions and dictates of various religions. In this study, religion-sensitive N-Calculator and P-Calculator approaches were used to determine the impact of religious dietary culture on the food N and P footprints of India in the major religious communities. Using 2013 data, the food N footprint of Hindus, Muslims, Christians, and Buddhists was 10.70, 11.45, 11.47, and 7.39 kg-N capita -1 year -1 (10.82 kg-N capita -1 year -1 was the national average), and the food P footprint was 1.46, 1.58, 1.04. and 1.58 kg-P capita -1 year -1 (1.48 kg-P capita -1 year -1 was the national average). The findings highlight the impact of individual choice on the N and P food footprints, and the importance of encouraging the followers of religion to follow a diet consistent with the food culture of that religion. The results of this study are a clear indication of the requirement for religion-sensitive analyses in the collecting of data pertinent to a particular country for use in making government policies designed to improve the recycling of food waste and the treatment of wastewater.

Background: Cardiac surgery with cardiopulmonary bypass (CPB) is associated with a high risk of postoperative acute kidney injury (AKI). Due to limitations of current diagnostic strategies, we sought to determine whether free hemoglobin (fHb) ratio (ie, levels of fHb at the end of CPB divided by baseline fHb) could predict AKI after on-pump cardiac surgery., Methods: This is a secondary analysis of a randomized controlled trial comparing the effect of nitric oxide (intervention) versus nitrogen (control) on AKI after cardiac surgery (NCT01802619). A total of 110 adult patients in the control arm were included. First, we determined whether fHb ratio was associated with AKI via multivariable analysis. Second, we verified whether fHb ratio could predict AKI and incorporation of fHb ratio could improve predictive performance at an early stage, compared with prediction using urinary biomarkers alone. We conducted restricted cubic spline in logistic regression for model development. We determined the predictive performance, including area under the receiver-operating-characteristics curve (AUC) and calibration (calibration plot and accuracy, ie, number of correct predictions divided by total number of predictions). We also used AUC test, likelihood ratio test, and net reclassification index (NRI) to compare the predictive performance between competing models (ie, fHb ratio versus neutrophil gelatinase-associated lipocalin [NGAL], N-acetyl-β-d-glucosaminidase [NAG], and kidney injury molecule-1 [KIM-1], respectively, and incorporation of fHb ratio with NGAL, NAG, and KIM-1 versus urinary biomarkers alone), if applicable., Results: Data stratified by median fHb ratio showed that subjects with an fHb ratio >2.23 presented higher incidence of AKI (80.0% vs 49.1%; P = .001), more need of renal replacement therapy (10.9% vs 0%; P = .036), and higher in-hospital mortality (10.9% vs 0%; P = .036) than subjects with an fHb ratio ≤2.23. fHb ratio was associated with AKI after adjustment for preestablished factors. fHb ratio outperformed urinary biomarkers with the highest AUC of 0.704 (95% confidence interval [CI], 0.592-0.804) and accuracy of 0.714 (95% CI, 0.579-0.804). Incorporation of fHb ratio achieved better discrimination (AUC test, P = .012), calibration (likelihood ratio test, P < .001; accuracy, 0.740 [95% CI, 0.617-0.832] vs 0.632 [95% CI, 0.477-0.748]), and significant prediction increment (NRI, 0.638; 95% CI, 0.269-1.008; P < .001) at an early stage, compared with prediction using urinary biomarkers alone., Conclusions: Results from this exploratory, hypothesis-generating retrospective, observational study shows that fHb ratio at the end of CPB might be used as a novel, widely applicable biomarker for AKI. The use of fHb ratio might help for an early detection of AKI, compared with prediction based only on urinary biomarkers., Competing Interests: Conflicts of Interest: See Disclosures at the end of the article., (Copyright © 2021 International Anesthesia Research Society.)

Amino-acids (AAs) are the exclusive source of nitrogen for cells. AAs result from the breakdown of food proteins and are absorbed by mucosa of the small intestine that act as a barrier to harmful materials. The quality of food proteins may differ, since it reflects content in Essential-AAs (EAAs) and digestibility but, until now, attention was paid mainly to the interaction between indigested proteins as a whole and microbiota. The link between microbiome and quality of proteins has been poorly studied, although these metabolic interactions are becoming more significant in different illnesses. We studied the effects of a special diet containing unbalanced EAAs/Non-EAAs ratio, providing excess of Non-EAAs, on the histopathology of gut epithelium and on the microbiome in adult mice, as model of qualitative malnutrition. Excess in Non-EAAs have unfavorable quick effect on body weight, gut cells, and microbiome, promoting weakening of the intestinal barrier. Re-feeding these animals with standard diet partially reversed the body alterations. The results prove that an unbalanced EAAs/Non-EAAs ratio is primarily responsible for microbiome modifications, not vice-versa. Therefore, treating microbiota independently by treating co-existing qualitative malnutrition does not make sense. This study also provides a reproducible model of sarcopenia-wasting cachexia like the human protein malnutrition.

This study aimed to identify its influence on pulmonary development and hemodynamics before bidirectional Glenn (BDG) anastomosis. This retrospective study involved 30 consecutive patients with univentricular hearts who underwent bilateral pulmonary artery banding (bil-PAB) and BDG anastomosis between September 2011 and September 2018 at our institution. These patients were classified into the nitrogen (N = 12) and no nitrogen (N = 18) groups according to their use of hypoxic gas therapy. Clinical echocardiographic, operative, and catheter examination data were reviewed. Nakata index was 131 (110-167) in the nitrogen group and 138 (88.4-161.7) in the no nitrogen group (P = 0.8). Pulmonary vascular resistance (PVR) was 1.50 (1.30-2.4) in the nitrogen group and 138 (88.4-161.7) in the no nitrogen group (P = 0.3). There was no statistical difference between the pulmonary development in both groups. In the subgroup of hypoplastic left heart syndrome patients, there was a statistical difference. RVEDV% of normal was 232.5 (215-239) in the nitrogen group and 201 (161-209) in the no nitrogen group (P = 0.03). Pulmonary development was not significantly different when hypoxic gas therapy was performed using nitrogen vs. not using nitrogen. However, RVEDV% of normal might be larger with hypoxic gas therapy using nitrogen. Although there is no obvious systemic atrioventricular valve regurgitation exacerbation with hypoxic gas therapy, it might be useful to prevent a decrease in PVR and may also affect bil-PAB and subclinical increase in systemic atrioventricular valve regurgitation.

Nitrogen (N) is a primary factor limiting leaf photosynthesis. However, the mechanism of high-N-driven inhibition on photosynthetic efficiency and photoprotection is still unclear in the shade-tolerant and N-sensitive species such as Panax notoginseng. Leaf chlorophyll (Chl) content, Ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco) activity and content, N allocation in the photosynthetic apparatus, photosynthetic performance and Chl fluorescence were comparatively analyzed in a shade-tolerant and N-sensitive species P. notoginseng grown under the levels of moderate nitrogen (MN) and high nitrogen (HN). The results showed that Rubisco content, Chl content and specific leaf nitrogen (SLN) were greater in the HN individuals. Rubisco activity, net photosynthetic rate (A net ), photosynthetic N use efficiency (PNUE), maximum carboxylation rate (V cmax ) and maximum electron transport rate (J max ) were lower when plants were exposed to HN as compared with ones to MN. A large proportion of leaf N was allocated to the carboxylation component under the levels of MN. More N was only served as a form of N storage and not contributed to photosynthesis in HN individuals. Compared with the MN plants, the maximum quantum yield of photosystem II (F v /F m ), non-photochemical quenching of PSII (NPQ), effective quantum yield and electron transport rate were obviously reduced in the HN plants. Cycle electron flow (CEF) was considerably enhanced in the MN individuals. There was not a significant difference in maximum photo-oxidation P 700+ (P m ) between the HN and MN individuals. Most importantly, the HN individuals showed higher K phase in the fast chlorophyll fluorescence induction kinetic curve (OJIP kinetic curve) than the MN ones. The results obtained suggest that photosynthetic capacity might be primarily inhibited by the inactivated Rubisco in the HN individuals, and HN-induced depression of photoprotection might be caused by the photodamage to the donor side of PSII oxygen-evolving complex.

The on-farm research network concept enables a group of farmers to test new agricultural management practices under local conditions with support from local researchers or agronomists. Different on-farm trials based on the same experimental design are conducted over several years and sites to test the effectiveness of different innovative management practices aimed at increasing crop productivity and profitability. As a larger amount of historical trial data are being accumulated, data of all the trials require analyses and summarization. Summaries of on-farm trials are usually presented to farmers as individual field reports, which are not optimal for the dissemination of results and decision making. A more practical communication method is needed to enhance result communication and decision making. R Shiny is a new rapidly developing technology for turning R data analyses into interactive web applications. For the first time for on-farm research networks, we developed and launched an interactive web tool called ISOFAST using R Shiny. ISOFAST simultaneously reports all trial results about the same management practice to simplify interpretation of multi-site and multi-year summaries. We used a random-effects model to synthetize treatment differences at both the individual trial and network levels and generate new knowledge for farmers and agronomists. The friendly interface enables users to explore trial summaries, access model outputs, and perform economic analysis at their fingertips. This paper describes a case-study to illustrate how to use the tool and make agronomic management decisions based on the on-farm trial data. We also provided technical details and guidance for developing a similar interactive visualization tool customized for on-farm research network. ISOFAST is currently available at https://analytics.iasoybeans.com/cool-apps/ISOFAST/., (© 2020 John Wiley & Sons Ltd.)

Cryotherapy is an ablation modality relying on freeze-thaw cycles to promote cell death through intracellular ice crystal formation, ischemia, and apoptosis. Currently, 2 different cryotherapy systems are available for esophageal use. The first is cryospray ablation, which involves repetitive applications of liquid nitrogen. The second system, cryoballoon ablation, freezes the esophageal mucosa with liquid nitrous oxide using a compliant balloon. To date, studies have shown that cryotherapy is an effective and safe method for eradicating Barrett's esophagus; however, larger prospective series with longer follow-up are warranted to determine the clinical utility of cryotherapy in the treatment of Barrett's esophagus., Competing Interests: Disclosure For C.N. Frederiks no conflict of interest was declared. M.I. Canto has received research grants for IRB-approved studies from Pentax Medical Corporation (America). B.L.A.M. Weusten has received financial support for IRB-approved studies from Pentax Medical Corporation (America)., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background Ventilation with the noble gas argon (Ar) has shown neuroprotective and cardioprotective properties in different in vitro and in vivo models. Hence, the neuroprotective effects of Ar were investigated in a severe, preclinically relevant porcine model of cardiac arrest. Methods and Results Cardiac arrest was ischemically induced in 36 pigs and left untreated for 12 minutes before starting cardiopulmonary resuscitation. Animals were randomized to 4-hour post-resuscitation ventilation with: 70% nitrogen-30% oxygen (control); 50% Ar-20% nitrogen-30% oxygen (Ar 50%); and 70% Ar-30% oxygen (Ar 70%). Hemodynamic parameters and myocardial function were monitored and serial blood samples taken. Pigs were observed up to 96 hours for survival and neurological recovery. Heart and brain were harvested for histopathology. Ten animals in each group were successfully resuscitated. Ninety-six-hour survival was 60%, 70%, and 90%, for the control, Ar 50%, and Ar 70% groups, respectively. In the Ar 50% and Ar 70% groups, 60% and 80%, respectively, achieved good neurological recovery, in contrast to only 30% in the control group ( P <0.0001). Histology showed less neuronal degeneration in the cortex ( P <0.05) but not in the hippocampus, and less reactive microglia activation in the hippocampus ( P =0.007), after Ar compared with control treatment. A lower increase in circulating biomarkers of brain injury, together with less kynurenine pathway activation ( P <0.05), were present in Ar-treated animals compared with controls. Ar 70% pigs also had complete left ventricular function recovery and smaller infarct and cardiac troponin release ( P <0.01). Conclusions Post-resuscitation ventilation with Ar significantly improves neurologic recovery and ameliorates brain injury after cardiac arrest with long no-flow duration. Benefits are greater after Ar 70% than Ar 50%.

Paeonia ostii is now being extensively planted for oil extraction in China, which is recognized as a single oil-use tree peony cultivar and commonly called 'Fengdan'. This study investigated the effects of nitrogen fertilizer on oil yield, fatty acid compositions and antioxidant activity of P. ostii. Oil yield (33.46 %), oleic acid (25.12 %), linoleic acid (29.21 %) and α-linolenic acid (43.12 %) reached the maximum at N450 treatment, with significant differences compared with other treatments (P<0.05). Furthermore, strong antioxidant activity with low DPPH IC50 value (19.43±1.91 μg mL -1 ) and large ABTS value (1216.53±30.21 μmol Trolox g -1 ) and FRAP value (473.57±9.11 μmol Trolox g -1 ) was also observed at N450. Palmitic acid (5.57 %) and stearic acid (2.02 %) reached a maximum at N375, but not significant with N450 (P<0.05). Nitrogen fertilizer could promote oil yield, fatty acid accumulation and antioxidant activity, and N450 (450 kg ha -1 ) is recommended as the optimum application for P. ostii., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Plant nitrogen (N) fertilization is known to affect disease; however, the underlying mechanisms remain mostly unknown. We investigated the impact of N supply on the Arabidopsis thaliana-Botrytis cinerea interaction. A. thaliana plants grown in low nitrate were more tolerant to all wild-type B. cinerea strains tested. We determined leaf nitrate concentrations and showed that they had a limited impact on B. cinerea growth in vitro. For the first time, we performed a dual RNA-Seq of infected leaves of plants grown with different nitrate concentrations. Transcriptome analysis showed that plant and fungal transcriptomes were marginally affected by plant nitrate supply. Indeed, only a limited set of plant (182) and fungal (22) genes displayed expression profiles altered by nitrate supply. The expression of selected genes was confirmed by quantitative reverse transcription PCR at 6 hr postinfection (hpi) and analysed at a later time point (24 hpi). We selected three of the 22 B. cinerea genes identified for further analysis. B. cinerea mutants affected in these genes were less aggressive than the wild-type strain. We also showed that plants grown in ammonium were more tolerant to B. cinerea. Furthermore, expression of the selected B. cinerea genes in planta was altered when plants were grown with ammonium instead of nitrate, demonstrating an impact of the nature of N supplied to plants on the interaction. Identification of B. cinerea genes expressed differentially in planta according to plant N supply unveils two novel virulence functions required for full virulence in A. thaliana: a secondary metabolite (SM) and an acidic protease (AP)., (© 2020 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Background: Experimental studies have investigated the effects of physical, psychological and pharmacological stressors (that induce state anxiety) on alcohol outcomes. However, no study has investigated the effects of state anxiety on alcohol outcomes, and the moderating role of drinking to cope (DTC) motives, using the 7.5% carbon dioxide (CO 2 ) challenge., Aims: We aimed to investigate the relationships between state anxiety and alcohol-related outcomes (primarily alcohol choice). We also explored whether DTC motives moderated these relationships., Methods: We conducted two experiments using the 7.5% CO 2 challenge (Studies 1 and 2) and an observational study (Study 3) ( n s = 42, 60 and 219, respectively), to triangulate findings., Results: In Study 1, experimentally induced state anxiety increased alcohol choice ( p < .001, η p 2  = .29). This finding was replicated in Study 2, but the effect was weaker ( p = .076, η p 2 = .06). Furthermore, DTC moderated the effect ( p = .013, η p 2 = . 11). However, in Study 3 there was no clear evidence of an association between naturally occurring state anxiety and alcohol choice ( b  = 0.05, p = .655), or a moderating role of DTC ( b = 0.01, p = .852)., Conclusions: Experimentally induced, but not naturally occurring, state anxiety increases alcohol choice, although state anxiety levels were lower in the non-manipulated sample.

The earth has been undergoing climate change, especially in recent years, driven by increasing concentration of atmospheric carbon dioxide (CO 2 ) and rising earth-surface temperature, which could reduce N allocation to Bt toxin for transgenic Bt crops (Bt crops), but the N fertilization is considered to be an effective method to enhance the C-N balance in Bt crops in the case of elevated CO 2 in future. DNA methylation not only in promoterregion but also in codingregion of transgene plays a critical role in transgene expression regulation and silencing of transgenic crops. Recent research has emphasized the risks of increased transgene silencing of Bacillus thuringiensis (Bt) rice under elevated CO 2 . In this study, the effects of elevated CO 2 (vs. ambient CO 2 ) on exogenous Bt toxins and transgene expression in promoterregion and codingregion of Bt rice during tillering stage (cv. HH1 expressing fused Cry1Ab/Cry1Ac) were evaluated under three nitrogen (N) fertilizer rate (1/4, 1 and 2 N levels). The aboveground and belowground biomass, and foliar Bt protein content of Bt rice were all significantly increased with the augmentation of N-fertilizer. And elevated CO 2 significantly increased belowground biomass, total soluble protein content, transgene methylation levels in promoterregion (P1), and in total of promoterregion(P1) and codingregion (P2 + P3) (i.e., P1 + P2 + P3) at 1 N level, and it also increased transgene methylation levels in codingregion (P2), and in total of promoterregion and codingregion (P1 + P2 + P3) at 2 N level. In addition, elevated CO 2 decreased foliar Bt protein content at 1 N level. The transgene methylation levels in promoterregion and codingregion were negatively correlated with Bt-transgene expression level. The methylation level of cytosines located at CG sites was higher than those at CHG and CHH sites in P1, P2 and P3 fragments regardless of the CO 2 or N-fertilizer level. The correlation of transgene mehtylation in promoterregion with transgene expression is even stronger than that in codingregion. These data indicate that N fertilization supply will increase the Bt toxin content in transgenic Bt rice, especially under elevated CO 2 .

The quality of rice grain is characterized by the component, structure and physicochemical properties of starch accumulated in endosperm cell. Nitrogen uptake strongly affects rice growth and starch development. In this study, Nangeng 9108 was used to investigated the accumulation of starch in different positions of the endosperm and physical properties of starch under nitrogen treatment of panicle initiation (PI) stage. Compared with the control group (CG), nitrogen treatment group (NTG) featured a higher number of grains per panicle and 1000-grain weight. Nitrogen treatment significantly increased starch accumulation among different regions during endosperm development, which was expressed as central endosperm cells > sub-aleurone cells of abdominal endosperm > sub-aleurone cells of dorsal endosperm. The amyloplast increased by constricting and budding-type division, generated a bead-like structure and derived some vesicles. The particle size of the starch granules obtained from the NTG was smaller and the apparent amylose content was lower than those of the CG, resulting in higher relative crystallinity. Nitrogen treatment promoted double helical components and provided a higher degree of order at short-rang scale for the starch granules. This study indicated that nitrogen significantly affected the accumulation and physicochemical properties of starch in the endosperm., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Abrupt drought-flood alternation is a frequent meteorological disaster during the summer in Southern China. The study of physiological and translation mechanisms of rice yield recovery after abrupt drought-flood alternation has great potential benefits in field production. Our results showed that yield recovery upon nitrogen (N) application after abrupt drought-flood alternation was due to the increase in effective panicle numbers per plant. The N application resulted in the regulation of physiological and biochemical as well as growth development processes, which led to a rapid growth recovery effect after abrupt drought-flood alternation stress in rice. Using ribosome profiling combined with RNA sequencing (RNA-seq) technology, the interactions between transcription and translation for N application after abrupt drought-flood alternation were analyzed. It was found that a small proportion of response genes were shared at the transcriptional and translational levels, that is, 14% of the expressed genes were upregulated and 6.6% downregulated. Further analysis revealed that the translation efficiency (TE) of the genes was influenced by their sequence characteristics, including their GC content, coding sequence length and normalized minimal free energy. Compared with the number of untranslated upstream open reading frames (uORFs), the increased number of translated uORFs promoted the improvement of TE. The TE of the uORFs for N application was lower than the control without N application after abrupt drought-flood alternation. This study characterizes the translational regulatory pattern in response to N application after abrupt drought-flood alternation stress., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Hybridoma and myeloma cell lines can be stored by slowly freezing cells in an appropriate solution of nutrients and a cryoprotectant such as glycerol or dimethyl sulfoxide (DMSO). In this protocol, cells are centrifuged at 4°C, resuspended in cold freezing solution (10% DMSO in FBS), and then transferred to an appropriate freezing vial. The vials are slowly frozen to -70°C in Styrofoam racks and then stored in liquid nitrogen (LN 2 ). Cells stored in LN 2 will remain viable for years. Once a frozen vial has been removed from LN 2 storage, it should be thawed as described, grown out into log phase, and refrozen., (© 2020 Cold Spring Harbor Laboratory Press.)

Learning more about the biodiversity and composition of arbuscular mycorrhizal fungi (AMF) under alternative agricultural management scenarios may be important to the sustainable intensification of switchgrass grown as a bioenergy crop. Using PacBio single-molecule sequencing and taxonomic resolution to the level of amplicon sequence variant (ASV), we assessed the effects of nitrogen amendment on AMF associating with switchgrass and explored relationships between AMF and switchgrass yield across three sites of various productivities in Wisconsin. Nitrogen amendment had little effect on AMF diversity metrics or community composition. While AMF ASV diversity was not correlated with switchgrass yield, AMF family richness and switchgrass yield had a strong, positive relationship at one of our three sites. Each of our sites was dominated by unique ASVs of the species Paraglomus brasilianum , indicating regional segregation of AMF at the intraspecific level. Our molecular biodiversity survey identified putative core members of the switchgrass microbiome, as well as novel clades of AMF, especially in the order Paraglomerales and the genus Nanoglomus Furthermore, our phylogenies unite the cosmopolitan, soil-inhabiting clade deemed GS24 with Pervetustaceae , an enigmatic family prevalent in stressful environments. Future studies should isolate and characterize the novel genetic diversity found in switchgrass agroecosystems and explore the potential yield benefits of AMF richness. IMPORTANCE We assessed the different species of beneficial fungi living in agricultural fields of switchgrass, a large grass grown for biofuels, using high-resolution DNA sequencing. Contrary to our expectations, the fungi were not greatly affected by fertilization. However, we found a positive relationship between plant productivity and the number of families of beneficial fungi at one site. Furthermore, we sequenced many species that could not be identified with existing reference databases. One group of fungi was highlighted in an earlier study for being widely distributed but of unknown taxonomy. We discovered that this group belonged to a family called Pervetustaceae , which may benefit switchgrass in stressful environments. To produce higher-yielding switchgrass in a more sustainable manner, it could help to study these undescribed fungi and the ways in which they may contribute to greater switchgrass yield in the absence of fertilization., (Copyright © 2020 Dirks and Jackson.)

Biochar-based controlled release nitrogen fertilizers (BCRNFs) have received increasing attention due to their ability to improve nitrogen-use efficiency (NUE) and increase crop yields. We previously developed a novel BCRNF, but its effects on soil microbes, NUE, and crop yields have not been reported. Therefore, we designed a pot experiment with five randomised treatments: CK (without urea and biochar), B (addition biochar without urea), B + U (biochar mixed urea), Urea (addition urea without biochar), and BCRNF (addition BCRNF), to investigate the effects of BCRNF on nitrifiers and denitrifiers, and how these impact nitrogen supply and NUE. Results of high-throughput sequencing revealed bacterial community groups with higher nutrient metabolic cycling ability under BCRNF treatment during harvest stage. Compared to Urea treatment, BCRNF treatment stimulated nitrification by increasing the copy number of the bacterial amoA gene and reducing nitrous oxide emission by limiting the abundance of nirS and nirK. Eventually, BCRNF successfully enhanced the yield (~ 16.6%) and NUE (~ 58.79%) of rape by slowly releasing N and modulating the abundance of functional microbes through increased soil nitrification and reduced denitrification, as compared with Urea treatment. BCRNF significantly improved soil NO 3 - , leading to an increase in N uptake by rape and NUE, thereby promoting rape growth and increasing grain yield.

Differently from the goal of fertilization in agricultural production to enhance the yield of seeds, the aim of present work focused on cadmium (Cd) phytoremediation is to increase the stem and leaf biomasses as much as possible due to they are the main organs of a hyperaccumulator extracting risk elements from soil. This experiment compared the effects of different nitrogen (N) application modes on Cd accumulation from soil by Solanum nigrum L. The results showed that stem and leaf biomasses of S. nigrum reached the maximum values with the N fertilizer application at Mode 4 (50 %: 40 %: 10 %, i.e. the front-heavy and back-light application of nitrogen fertilizer). In particular, application of N as (NH 4 ) 2 SO 4 and CH 4 N 2 O produced the greatest increases in S. nigrum biomasses compared to the control (CK 2 ) with no N fertilizer. N concentration of S. nigrum and N fertilizer productivity under its different treatments were consistent with the above results. Similarly, Cd capacities were the highest in shoots of S. nigrum in Mode 4 either due to their Cd concentration without affected by N fertilizers. Thus, Mode 4 N application showed very important scientific merit for effective Cd phytoextraction in the real conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Effects of the hyperbaric environment on the autonomic nervous system (ANS) in recreational divers are not firmly settled. Aim of this exploratory study was to (1) assess ANS changes during scuba diving via recordings of electrocardiograms (ECG) and to (2) study whether nitrox40 better improves sympathovagal balance over air. 13 experienced divers (~40yrs) performed two open-water dives each breathing either air or nitrox40 (25m/39min). 3-channel ECGs were recorded using a custom-made underwater Holter-monitor. The underwater Holter system proved to be safe. Air consumption exceeded nitrox40 consumption by 12% (n = 13; p < 0.05). Both air and nitrox40 dives reduced HR (10 vs 13%; p < 0.05). The overall HRV (pNN50: 82 vs 126%; p < 0.05) and its vagal proportion (RMSSD: 33 vs 50%; p < 0.05) increased during the dive. Moreover, low (LF: 61 vs 47%) and high (HF: 71 vs 140%) frequency power were increased (all p < 0.05), decreasing the ratio of LF to HF (22 vs 34%). : Conventional open-water dives distinctly affect the ANS in experienced recreational divers, with sympathetic activation less pronounced than vagal activation thereby improving the sympathovagal balance. Nitrox40 delivered two positive results: nitrox40 consumption was lower than air consumption, and nitrox40 better improved the sympathovagal balance over air.

Many fertilization models have been created to scientifically determine the amount of fertilization. With the same purpose, we constructed a nitrogen (N) application model, the leaf value model, which can make N fertilizer decisions in a timely, fast and nondestructive manner during rice planting. However, only one area (A1, Jiuzhou Town, Xixiu District, Guizhou Province) and one cultivar (Qyou6) were involved in the construction of the leaf value model. Its stability and applicability could not be well evaluated. Thus, we chose another area (A2, Jiuzhou Town, Huangping County, Guizhou Province) in Guizhou Province and carried out the experiment by using four cultivars (Nie5you5399, Qyou6, Yixiangyou2115 and Zhongzheyou8) for the leaf value model construction. Compared with the average value of apparent total N uptake (Nz) obtained in 2 years in the A1 area, that in the Qyou6 leaf value model in the A2 area increased by 12%, reaching 635.72 kg ha-1, whereas the corresponding target yield changed slightly, reaching 10,999.90 kg ha-1. Simultaneously, the linear relationship between several good SPAD value-derived indexes (Ys) and apparent N supply of the field (Nx) was still significant or extremely significant in the Qyou6 leaf value model. Compared with the A1 area, it slightly differed, and the R2 of SPADL1 was higher than that of SPADL3×L4/mean. In the leaf value model of the other three cultivars, the relationship between yield and Nx and that between Ys and Nx were significant or extremely significant. The Nz of Yixiangyou2115 and Zhongzheyou8 (618.33 and 617.76 kg ha-1) were close to that of Qyou6 and the corresponding target yields were 10313.36 and 10301.99 kg ha-1, respectively. The Nz and target yield of Nie5you5399 were lowest at 546.63 and 10680.24 kg ha-1, respectively. In general, this study showed that relationships used in the construction of leaf value model had certain stability and applicability to difference areas and cultivars. The leaf value model can be considered in N fertilizer decision-making of rice planting management., Competing Interests: The authors have declared that no competing interests exist.

The present study aimed to evaluate the degradation dynamics of high-quality tropical forage according to the levels of nitrogen supplementation. In this study, 4 rumen-fistulated cows with a body weight of 653 ± 47 kg were used. These animals were distributed in a 4 × 4 Latin square experimental design. Cows were kept under a grazing regime in an area formed by Panicum maximum Jacq. cv. Tanzania. These animals received 4 levels of crude protein (CP) in the supplements (122, 142, 162, and 180 g/kg of CP in DM). The degradability of dry matter (DM), crude protein (CP), and neutral detergent fiber (NDF) was determined by in situ incubation of forage samples at 0, 6, 12, 24, 48, 72, 96, and 120 h. Both the concentration of ammonia nitrogen (NH 3 -N) and the microbial protein synthesis were affected by CP levels of the supplements (P < 0.05). There was a significant difference (P < 0.05) for fraction b, potential degradability, and effective degradability of DM and NDF, and for the c degradation rate of CP of Tanzania grass which had the highest values at 122 g/kg of CP. Overall, the greatest degradation of nutrients from Tanzania grass occurred in the diet with 122 g/kg of CP and 604 g/kg of NFC in dairy cattle supplements that resulted in the highest rates of nitrogen utilization by rumen microbes.

Molecular hydrogen is thought to have an inhibitory effect on oxidative stress, thereby attenuating the onset and progression of various diseases including cardiovascular disease; however, few reports have assessed the preventive effect of constitutive inhalation of hydrogen gas on of vascular remodeling. Here, we investigated the effect of constitutive inhalation of hydrogen gas on vascular neointima formation using a cuff-induced vascular injury mouse model. After constitutive inhalation of compressed hydrogen gas (O2 21%, N2 77.7%, hydrogen 1.3%) or compressed air only (O2 21%, N2 79%) by C57BL/6 mice for 2 weeks from 8 weeks of age in a closed chamber, inflammatory cuff injury was induced by polyethylene cuff placement around the femoral artery under anesthesia, and hydrogen gas administration was continued until sampling of the femoral artery. Neointima formation, accompanied by an increase in cell proliferation, was significantly attenuated in the hydrogen group compared with the control group. NADPH oxidase NOX1 downregulation in response to cuff injury was shown in the hydrogen group, but the expression levels of NADPH oxidase subunits, p40phox and p47phox, did not differ significantly between the hydrogen and control groups. Although the increase in superoxide anion production did not significantly differ between the hydrogen and control groups, DNA damage was decreased as a result of reduction of reactive oxygen species such as hydroxyl radical (⋅OH) and peroxynitrite (ONOO-) in the hydrogen group. These results demonstrate that constitutive inhalation of hydrogen gas attenuates vascular remodeling partly via reduction of oxidative stress, suggesting that constitutive inhalation of hydrogen gas at a safe concentration in the living environment could be an effective strategy for prevention of vascular diseases such as atherosclerosis., Competing Interests: The authors have declared that no competing interests exist.

Background: The Eurasian steppe is an important vegetation type characterized by cold, arid and nitrogen poor conditions. At the Eastern edge, including in the Songnen grassland, the vegetation is dominated by Leymus chinensis (henceforth L. chinensis) and is increasing threatened by elevated anthropogenic nitrogen deposition. L. chinensis is a perennial grass that mainly reproduces vegetatively and its sexual reproduction is limited. However, sexual reproduction plays an important role influencing colonization after large disturbances. To develop an understanding of how elevated nitrogen deposition changes the plant community structure and functioning we need a better understanding how sexual reproduction of L. chinensis changes with nitrogen enrichment. Here we report on a field experiment where we added 10 g N m - 2  yr - 1 and examined changes in seed traits, seed germination and early seedling growth., Results: Nitrogen addition increased seed production by 79%, contributing to this seed increases were a 28% increase in flowering plant density, a 40% increase in seed number per plant and a 11% increase in seed weight. Seed size increased with a 42% increase in large seeds and a 49% decrease in the smallest seed size category. Seed germination success improved by 10% for small seeds and 18% for large seeds. Combined, the increased in seed production and improved seed quality doubled the potential seed germination. Subsequent seedling above and below-ground biomass also significantly increased., Conclusions: All aspects of L. chinensis sexual reproduction increased with nitrogen addition. Thus, L. chinensis competitive ability may increase when atmospheric nitrogen deposition increases, which may further reduce overall plant diversity in the low diversity Songnen grasslands.

Soil extracellular enzymes play a significant role in the N mineralization process. However, few studies have documented the linkage between enzyme activity and the microbial community that performs the function. This study examined the effects of inorganic and organic N fertilization on soil microbial communities and their N mineralization functions over 4 years. Soils were collected from silage corn field plots with four contrasting N treatments: control (no additional N), ammonium sulfate (AS; 100 and 200 kg of N ha -1 ), and compost (200 kg of N ha -1 ). Illumina amplicon sequencing was used to comprehensively assess the overall bacterial community (16S rRNA genes), bacterial ureolytic community ( ureC ), and bacterial chitinolytic community ( chiA ). Selected genes involved in N mineralization were also examined using quantitative real-time PCR and metagenomics. Enzymes (and marker genes) included protease ( npr and sub ), chitinase ( chiA ), urease ( ureC ), and arginase ( rocF ). Compost significantly increased diversity of overall bacterial communities even after one application, while ammonium fertilizers had no influence on the overall bacterial communities over four seasons. Bacterial ureolytic and chitinolytic communities were significantly changed by N fertilization. Compost treatment strongly elevated soil enzyme activities after 4 years of repeated application. Functional gene abundances were not significantly affected by N treatments, and they were not correlated with corresponding enzyme activities. N mineralization genes were recovered from soil metagenomes based on a gene-targeted assembly. Understanding how the structure and function of soil microbial communities involved with N mineralization change in response to fertilization practices may indicate suitable agricultural management practices that improve ecosystem services while reducing negative environmental consequences. IMPORTANCE Agricultural N management practices influence the enzymatic activities involved in N mineralization. However, specific enzyme activities do not identify the microbial species directly involved in the measured process, leaving the link between the composition of the microbial community and the production of key enzymes poorly understood. In this study, the application of high-throughput sequencing, real-time PCR, and metagenomics shed light on how the abundance and diversity of microorganisms involved in N mineralization respond to N management. We suggest that N fertilization has significantly changed bacterial ureolytic and chitinolytic communities., (Copyright © 2020 American Society for Microbiology.)

Preclinical imaging studies of fetal hemodynamics require anesthesia to immobilize the animal. This may induce cardiovascular depression and confound measures under investigation. We compared the impact of four anesthetic regimes upon maternal and fetal blood gas and hemodynamics during baseline periods of normoxia, and in response to an acute hypoxic challenge in pregnant sheep. Merino ewes were surgically prepared with maternal and fetal vascular catheters and a fetal femoral artery flow probe at 105-109 days gestation. At 110-120 days gestation, ewes were anesthetized with either isoflurane (1.6%), isoflurane (0.8%) plus ketamine (3.6 mg·kg -1 ·h -1 ), ketamine (12.6 mg·kg -1 ·h -1 ) plus midazolam (0.78 mg·kg -1 ·h -1 ), propofol (30 mg·kg -1 ·h -1 ), or remained conscious. Following 60 min of baseline recording, nitrogen was administered directly into the maternal trachea to displace oxygen and induce maternal and thus fetal hypoxemia. During normoxia, maternal PaO 2 was ~30 mmHg lower in anesthetized ewes compared to conscious controls, regardless of the type of anesthesia (p < .001). There was no effect of anesthesia on fetal mean arterial blood pressure (MAP; p > .05), but heart rate was 32 ± 8 bpm lower in fetuses from ewes administered isoflurane (p = .044). During maternal hypoxia, fetal MAP increased, and peripheral blood flow decreased in all fetuses except those administered propofol (p < .05). Unexpectedly, hypoxemia also induced fetal tachycardia regardless of the anesthetic regime (p < .05). These results indicate that despite maternal anesthesia, the fetus can mount a cardiovascular response to acute hypoxia by increasing blood pressure and reducing peripheral blood flow, although the heart rate response may differ from when no anesthesia is present., (© 2020 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

Aims and Background: Spirulina (Arthrospira) platensis (SP) microalgae were cultured in Zarrouk Medium (ZM), containing three nitrogen concentrations (N-limited, N-optimal and Nrich medium) in ten liter-photo-bioreactor (10 L PBR) for 15-days, in order to study changes in lipid compounds (total carotenoids and total lipids and their effect on fatty acid profile). Based on US patent, the yield of bioactive compounds (such as gamma-linolenic acid GLA, C18:3) extracted from microalgae biomass, mainly depends on the extraction processes (1). GLA has much attention with respect to its therapeutic properties such as its ability to decrease blood cholesterol levels., Methods: The impact of the addition of N in cultures of S. platensis in terms of growth, biomasses and induced lipid compounds (total carotenoids and total lipid contents and its fatty acid profile), as well as the Sonication (SON) and Microwave (MIC) process as aiding techniques for lipid extraction compared with a Cold Condition (COL), was examined. GC/MS method was used to determine the fatty acid profile of lipid extract of SP cultures., Results: In all S. platensis tested culture, the SP was growing successfully, with varying degrees. In N-rich media, the highest cell growth rate and biomass yield were obtained compared with that recorded in other cultures. Under an N-limited condition, SP had higher Total Carotenoids (TCAR, 45.54 mg/g dw) and total lipid contents (TL, 29.51%± 1.92 g/100g dw) compared with that recorded either in N-rich (11.2 mg/g dw) or in N-optimal (6.23 mg/g dw) cultures. Thus, SP copes with the N -stress by altering the metabolic pathways towards inducing lipid biosynthesis. To maximize the TL and TCAR extraction yields, from N-limited cultures, a set of operating process was applied including the Sonication (SON) and Microwave (MIC), which were used as aiding techniques for lipid extraction compared with the Cold Condition (COL) techniques. The results showed that the extraction efficiency of the S. platensis TL increased in the following order: MIC (29.51%± 1.92) > SON (25.46% ± 1.65> COL (20.43% ±1.43). In a comparative study for its fatty acid profiles (FAPs) among all SP cultures, lipids were analyzed by GC/MS. The predominant fatty acids (>10%, of total FA) were found to be myristic acid (C14:0, MA), palmitic acid (C16:0, PA) and oleic acid (C18:1)., Conclusion: The study concluded that the N-limited condition was found to have a strong influence on biomass dry weight and lipid contents and total carotenoids in SP cells compared to either Nrich or N-optimal conditions. The use of sonication and the microwave techniques lead to a great increase in the extraction of lipid contents and in high amount Polyunsaturated Fatty Acids (PUFAs) in N-limited cultures, in particular, the omega-6 (ω 6) and omega-3 (ω 3) of the essential C18 fatty acids. It seems that the SP rich in lipid content with a high amount of GLC produced under nitrogen limitation in PBR conditions can be used as a food additive or as a nutritional supplement., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The ruminant is able to transform plant fibres and non-protein nitrogen (NPN) into edible foods for human consumption. In an effort towards improving our understanding of this process, we sought to challenge convention and examine how the source, amount and site of NPN delivery in the gastrointestinal tract of the ruminant may affect fibre digestibility, rumen stability and N metabolism. In the first study presented here, we used four ruminally cannulated non-lactating heifers in a Latin square design to infuse 59 g/days of N in the form of ammonia (A) or urea (U) into either the rumen (R) or the abomasum (A). We found that intake was higher (p = .03) for animals receiving supplementary NPN as urea compared with ammonia. In addition, abomasally infused urea (UA) improved fibre fermentation by 9.4% (p = .05) and decreased ruminal pH fluctuations (lower slope in the cumulative pH parameters, p = .04) when compared with the same amount of urea infused ruminally (UR). In a second study, using the same group of heifers, we infused 50 or 150 g/day of urea into the rumen (UR50 and UR150) or 150 g of urea in the abomasum (UA150) or 50 g in the rumen and 100 g in the abomasum (URA150). Fibre digestion was improved by 4% (p = .02) when the same dose of urea was infused into the abomasum compared with the rumen, while estimated microbial protein production and N efficiency were not different between a low rumen dose and high post-ruminal dose of urea. Collectively, these studies provide insight into the viability of post-ruminal NPN supplementation as a strategy to improve fibre digestion and NPN inclusion in the ruminant diet., (© 2019 Blackwell Verlag GmbH.)

Culturing slowly growing tree seedlings is a potential approach for managing the conflict between the increasing demand for ornamental stock and the decreasing area of farmlands due to urbanization. In this study, Buddhist pine (Podocarpus macrophyllus [Thunb.] D. Don) seedlings were raised in multishelves with light-emitting diode lighting in the spectrum of 17:75:8 (red:green:blue) at 190-320 μmol m-2 s-1 with controlled temperature and relative humidity at 19.5°C and 60%, respectively. Seedlings were fed by exponential fertilization (EF) (nitrogen [N]-phosphorus [P]2O5-K2O, 10-7-9) at eight rates of 0 (control), 20 (E20), 40 (E40), 60 (E60), 80 (E80), 100 (E100), 120 (E120), and 140 (E140) mg N seedling-1 for four months through 16 fertilizer applications. The nutritional responses of Buddhist pine seedlings can be identified and classified into various stages in response to increasing doses, up to and over 120 N seedling-1. Morphological traits, i.e., the green color index and leaf area (LA) obtained by digital analysis and the fine root growth, all remained constant in response to doses that induced steady nutrient loading. LA had a positive relationship with most of the nutritional parameters. A dose range between 60 and 120 mg N seedling-1 was recommended for the culture of Buddhist pine seedlings. At this range of fertilizer doses, measuring the leaf area through digital scanning can easily and rapidly indicate the inherent nutrient status of the seedlings., Competing Interests: NO authors have competing interests.

Global change apart from ecosystem processes also influences the community structure of key organisms, such as arbuscular mycorrhizal fungi (AMF). We conducted a 3-year experiment where we suppressed with benomyl mycorrhiza to understand how AMF alter the plant community structure under warming and nitrogen (N) addition. The elemental content and foliar tissue stoichiometry of the dominant species Leymus chinensis and the subordinate species Puccinellia tenuiflora were studied along with soil nutrient stoichiometries. Overall, N addition enhanced plant N: phosphorus (P) ratios at a greater level than experimental warming did. Under global change conditions, AMF symbionts significantly increased soil available P concentrations, promoted plant P absorption and decreased the plant N:P ratios. AMF alleviate P limitation by reducing plant N:P ratios. Our results highlight that the negative influence of global change on plant productivity might cancel each other out through the additive effects of AMF and that global change will increase the dependency of plants on their mycorrhizal symbionts., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study examined the physiological effects of different amounts of nitrogen (N) supplementation (0 to 2.72 kg/year) on the citrus cultivar Huangguogan ( Citrus reticulata × Citrus sinensis ). Root activity, chlorophyll content, and fruit quality were measured, and the activities of the antioxidant enzymes superoxide dismutase (SOD), guaiacol peroxidase (POD), and catalase (CAT), and the contents of malondialdehyde (MDA) and soluble protein in root, leaf, and fruit tissues were examined at different developmental stages. Root activity, chlorophyll content, fruit quality, antioxidant enzyme activity, MDA content, and soluble protein content increased in plants treated with an appropriate amount of N. Both excessive N and N deficiency decreased the content of MDA and the activities of antioxidant enzymes. Application of 1.36-1.81 kg N/year is suggested for citrus fertilization and the lower end of this range is recommended for minimizing environmental impact and production cost.

This study firstly introduced a silicone rubber membrane (SRM) into microbial fuel cell (MFC) for passive oxygen supply to simultaneously remove phenol and nitrogen from synthetic coke-oven wastewater diluted with seawater. Passive oxygen transport with biofilm on the membrane was improved by ~ 18-fold in comparison with the one without a biofilm. In addition, although the oxygen supply was passive, nitrification accounted for 34% of those aeration conditions. It was also found that silicone rubber membrane can control NO 2 - -N and/or NO 3 - -N production. A dual-chamber MFC treating the synthetic coke-oven wastewater achieved a maximum power density of 54 mW m -2 with a coulombic efficiency of 2.7%. We conclude that silicone rubber membrane is effective for sustainable coke-oven wastewater treatment in MFCs.

Ammonium pulse attributed to runoff of urban surface and agriculture following heavy rain is common in inland aquatic systems and can cause profoundly effects on the growth of macrophytes, especially when combined with low light. In this study, three patterns of NH4-N pulse (differing in magnitude and frequency) were applied to examine their effects on the growth of three submersed macrophytes, namely, Myriophyllum spicatum, Potamogeton maackianus, and Vallisneria natans, in terms of biomass, height, branch/ramet number, root length, leaf number, and total branch length under high and low light. Results showed that NH4-N pulse caused negative effects on the biomass of the submerged macrphytes even on the 13th day after releasing NH4-N pulse. The negative effects on M. spicatum were significantly greater than that on V. natans and P. maackianus. The effects of NH4-N pulse on specific species depended on the ammonium loading patterns. The negative effects of NH4-N pulse on P. maackianus were the strongest at high loading with low frequency, and on V. natans at moderate loading with moderate frequency. For M. spicatum, no significant differences were found among the three NH4-N pulse patterns. Low light availability did not significantly aggregate the negative effects of NH4-N pulse on the growth of the submersed macrophytes. Our study contributes to revealing the roles of NH4-N pulse on the growth of aquatic plants and its species specific effects on the dynamics of submerged macrophytes in lakes., Competing Interests: The authors have declared that no competing interests exist.

Rapid hydrolysis of urea results in further fertilization frequency and excessive nitrogen (N) input. A modified urea, dimethylolurea (DMU), was synthesized in this study. The structure of the sample was characterized by Fourier transform infrared and nuclear magnetic resonance analysis, manifesting the formation of DMU. N release investigation confirmed that DMU enabling provided a gradual N supply. The N leaching experiment indicated that increasing the applied DMU significantly reduced the NH 4 + -N, NO 3 - -N, and total N leaching, compared with urea application alone. The application effect on maize and wheat was evaluated. The results revealed that singly applied DMU with 100% or 80% N input, irrespective of the amount, promoted crop yield and agronomic characteristic and N use efficiency (NUE) of maize and wheat, beyond urea with two split applications at the recommended rate. Thus, the potential availability of DMU was proven; this could be widely used in agricultural fields as a slow-release fertilizer.

Objective: To assess effects of nitrogen and helium on efficacy of an alveolar recruitment maneuver (ARM) for improving pulmonary mechanics and oxygen exchange in anesthetized horses., Animals: 6 healthy adult horses., Procedures: Horses were anesthetized twice in a randomized crossover study. Isoflurane-anesthetized horses in dorsal recumbency were ventilated with 30% oxygen and 70% nitrogen (treatment N) or heliox (30% oxygen and 70% helium; treatment H) as carrier gas. After 60 minutes, an ARM was performed. Optimal positive end-expiratory pressure was identified and maintained for 120 minutes. Throughout the experiment, arterial blood pressures, heart rate, peak inspiratory pressure, dynamic compliance (C dyn ), and Pao 2 were measured. Variables were compared with baseline values and between treatments by use of an ANOVA., Results: The ARM resulted in significant increases in Pao 2 and C dyn and decreases in the alveolar-arterial gradient in the partial pressure of oxygen in all horses. After the ARM and during the subsequent 120-minute phase, mean values were significantly lower for treatment N than treatment H for Pao 2 and C dyn . Optimal positive end-expiratory pressure was consistently 15 cm H 2 O for treatment N, but it was 10 cm H 2 O (4 horses) and 15 cm H 2 O (2 horses) for treatment H., Conclusions and Clinical Relevance: An ARM in anesthetized horses might be more efficacious in improving Pao 2 and C dyn when animals breathe helium instead of nitrogen as the inert gas.

Experiments were conducted to estimate daily N maintenance requirements (NMRs) and the genetic potential for daily N retention (NR max T) of pullets in growth phase. Three nitrogen balance trials were conducted, and a total of 48 Hy-line W-36 pullets were used in each trial in age periods (starter: 14-28, grower: 56-70 and developer: 98-112 days). The treatments consisted of six graded levels of nitrogen in the diets (L1 = 8, L2 = 16, L3 = 24, L4 = 32, L5 = 40 and L6 = 48 g N/kg of feed), formulated using the dilution technique. The regression analyses between nitrogen intake and nitrogen excretion were performed to fit the exponential function and to determine the NMR. The daily NMRs that were estimated at 294, 331 and 355 mg/BW kg 0.67 for the initial, grower and developer periods, respectively, were applied for further calculation of NR max T as the threshold value of the function between N intake and daily N balance. The NR max T was estimated by a statistical procedure following several iteration steps by the Levenberg-Marquardt algorithm until the sum of the squares of the residual was minimized. The NR max T was estimated at 3,200, 2,633 and 1,826 mg/BW kg 0.67 for starter, grower and developer periods respectively. The determined model parameters were the precondition for modelling of the amino acid requirement based on an exponential N-utilization model and depended on performance and dietary amino acid efficiency. This procedure will be further developed and applied in the subsequent study., (© 2019 Blackwell Verlag GmbH.)

Introduction: Cognitive impairment related to inert gas narcosis (IGN) is a threat to diving safety and operations at depth that might be reduced by using enriched air nitrox (EANx) mixtures. Using critical flicker fusion frequency (CFFF), a possible early detection of cognitive abilities/cerebral arousal impairment when breathing different oxygen (O2) fractions was investigated., Methods: Eight male volunteers performed, in random order, two dry chamber dives breathing either air or EANx40 (40% O₂-60% nitrogen) for 20 minutes (min) at 0.4 MPa. Cognition and arousal were assessed before the dive; upon arrival at 0.4 MPa; after 15 min exposure at 0.4 MPa; on surfacing and 30 min post-dive using behavioural computer-based testing psychology experiment building language (PEBL) and by CFFF while continuously recording brain oxygenation with near-infrared spectroscopy., Results: In both breathing conditions, CFFF and PEBL demonstrated a significant inverse correlation (Pearson r of -0.90, P < 0.0001), improved cognitive abilities/cerebral arousal occurred upon arrival at 0.4 MPa followed by a progressive deterioration. Initial brain activation was associated with a significant increase in oxyhaemoglobin (HbO2) and a simultaneous decrease of deoxyhaemoglobin (HHb). The magnitude of the changes was significantly greater under EANx (P = 0.038)., Conclusions: Since changes were not related to haemodynamic variables, HbO₂ and HHb values indicate a significant, O₂-dependent activation in the prefrontal cortex. Owing to the correlation with some tests from the PEBL, CFFF could be a convenient measure of cognitive performance/ability in extreme environments, likely under the direct influence of oxygen partial pressure, a potent modulator of IGN symptoms., (Copyright: This article is the copyright of the authors who grant Diving and Hyperbaric Medicine a non-exclusive licence to publish the article in electronic and other forms.)

We hypothesized that heart rate variability (HRV) can be used as a physiological monitor of exposures to hypercapnia. HRV was analyzed in 15 male subjects breathing air and then air plus 4%, 5% or 6% carbon dioxide (CO2). A progressive elevation in randomness of HRV best depicted as a divergence of data on Poincar é plots of self-similarity occurred as CO2 increased, and resolved when returning sd to ambient conditions. Results indicate that HRV may be useful as a prompt (⟨ 2 minute) early warning of hypercapnia that occurs prior to overt symptomatology., Competing Interests: The authors of this paper declare no conflicts of interest exist with this submission., (Copyright© Undersea and Hyperbaric Medical Society.)

Soil amendment with biochar has received increased attention because of its potential to i) sequester carbon and ii) reduce N 2 O emission when applied to N fertilised soils. To study the effect of biochar origin on greenhouse gas emission in two contrasting soil types, we used a robotized continuous flow incubation system and δ 13 C stable isotope approach to compare four biochar types (feed stock: olive mill, corn cob, pistachio shell, cotton stalk) in an alkaline clay soil and two selected biochar types (feed stock: olive mill and corn cob) in an acidic sandy soil. Furthermore, high-throughput sequencing of 16S rRNA genes was performed at the end of the incubation to investigate the effect of different biochars on bacterial community structure in the two different soils. In the alkaline clay soil, all biochar types in conjunction with N fertiliser decreased CO 2 emissions up to 12% compared to the N added control treatment causing negative priming, whereas no significant effect of biochar addition on N 2 O emissions was observed. In contrast, application of olive mill biochar to the acidic sandy soil significantly increased soil pH, CO 2 , and N 2 O fluxes, whereas no significant effect of corn cob biochar addition was observed. There was a significant linear relationship between the biochar induced increase in soil pH and the biochar induced increase in soil born N 2 O emission. Additionally, we detected a clear variation in bacterial community structure in the acidic sandy soil (phyla Acidobacteria, Nitrospirare, and Arthrobacter) with the olive mill biochar addition. Overall, the amendment of different biochars failed to mitigate N 2 O emissions in both soil types when mineral fertiliser was added. Furthermore, amendment of olive mill biochar stimulated both N 2 O and CO 2 emissions in the low pH sandy soil and altered the bacterial community structure, which was possibly related to its liming effect., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Field experiments were carried out in 2016 and 2017 to study the influence of timing the application of nitrogen fertilizer (N-fertilizer) on the susceptibility of potatoes to early blight. We hypothesized that potatoes that receive N-fertilizer as split applications or a one-time application at emergence will have a higher N content and be less susceptible to early blight than those that receive N-fertilizer as a one-time application before planting. N-fertilizer was applied either as a one-time application before planting or at emergence and as split applications., Results: Potatoes that did not receive N-fertilizer were more susceptible to early blight [high area under the disease progress curve (AUDPC)] than those that did receive N-fertilizer (low AUDPC). The timing of N-fertilizer application had a significant effect on the susceptibility of potatoes to early blight. Potatoes that received the entire quantity of N-fertilizer at emergence or as split applications had higher N contents and were less susceptible to early blight than those that received the entire quantity of N-fertilizer before planting. Potatoes that did not receive N-fertilizer had a markedly lower starch yield than potatoes that received N-fertilizer. Potatoes that received the entire quantity of N-fertilizer before planting had a lower starch yield than potatoes that received the entire quantity of N-fertilizer at emergence or as split applications., Conclusion: N-fertilizer applied as a one-time application at emergence or as a split application was the best treatment to ensure both high N content and lower attack of early blight, which confirms our hypothesis. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Changes in nutrient dose have dramatic effects on gene expression and development. One outstanding question is whether organisms respond to changes in absolute nutrient amount (moles) vs. its concentration in water (molarity). This question is particularly relevant to plants, as soil drying can alter nutrient concentration, without changing its absolute amount. To compare the effects of amount vs. concentration, we expose rice to a factorial matrix varying the dose of nitrogen (N) and water (W) over a range of combinations, and quantify transcriptome and phenotype responses. Using linear models, we identify distinct dose responses to either N-moles, W-volume, N-molarity (N/W), or their synergistic interaction (N×W). Importantly, genes whose expression patterns are best explained by N-dose and W interactions (N/W or N×W) in seedlings are associated with crop outcomes in replicated field trials. Such N-by-W responsive genes may assist future efforts to develop crops resilient to increasingly arid, low nutrient soils.