Phosphoenolpyruvate Carboxylase Research Articles

Joint expression of Zmpepc, Zmppdk, and Zmnadp-me is more efficient than expression of one or two of those genes in improving the photosynthesis of Arabidopsis

This study assessed the effects of seven combinations of maize (Zea mays) genes phosphoenolpyruvate carboxylase (pepc), pyruvate phosphate dikinase (ppdk), and NADP-malic enzyme (nadp-me), on the photosynthesis of Arabidopsis. The photosynthetic rate, carboxylation efficiency, and shoot-dry-weight of Zmpepc (PC), Zmpepc + Zmppdk (PCK), Zmpepc + Zmnadp-me (PCM), and Zmpepc + Zmppdk + Zmnadp-me (PCKM) were significantly higher than those of the control wild-type (WT), with a trends to be PCKM > PCK > PC and PCM > WT. This indicated that Zmpepc was a prerequisite for improved photosynthetic performance; Zmppdk had a positive effect on Zmpepc, and the triple gene combination had the most significant synergistic effects. PCKM significantly enhanced activity of photosystem (PS)II (K, J phase) and PSI, light energy absorption (ABS/CSm) and conversion (TRo/ABS), and electron transfer (ETo/TRo). PCKM up-regulated 18 photosynthesis-related proteins, among which, 11 were involved in light reaction resulting in improved light-energy absorption and conversion efficiency, electron transfer, activity and stability of PSII and PSI, and the ATP and NADPH production. The remaining seven proteins were involved in dark reaction. The up-regulation of these proteins in PCKM improved the coordinated operation of light and dark reaction, increasing the photosynthesis and dry weight ultimately. These results also provide a promising strategy for the genetic improvement of the photosynthetic performance of C3 crops by inserting major C4 photosynthetic genes.

Erratum for Tong et al., "Gene Dispensability in Escherichia coli Grown in Thirty Different Carbon Environments".

Volume 11, no. 5, e02259-20, 2020, [https://doi.org/10.1128/mBio.02259-20][1]. On PDF page 1, in the abstract, in the sentence “From our comparisons to the model, we found that, unexpectedly, phosphoenolpyruvate carboxylase ( ppc ) was required for robust growth in most carbon sources other than

Effect of drought and heat stresses on photosynthesis, pigments, and xanthophyll cycle in alfalfa (Medicago sativa L.)

Climate changes severely influence photosynthetic activity of plants. Studies on the combined effect of drought and heat stress on photosynthesis, pigments, and xanthophyll cycle particularly in alfalfa (Medicago sativa L.) are limited. Thus, this study investigated the combined effects of drought and heat stress in two alfalfa varieties ('Deqin' and 'Algonguin'). Results revealed that the values of the net photosynthetic rate demonstrated a significantly decreasing tendency under stresses. The effect of stresses on ribulose 1,5-bisphosphate carboxylase activity in both varieties showed a trend of first declining and then rising. Meanwhile, phosphoenolpyruvate carboxylase activity showed an increasing tendency. The intensification of stresses led to reduced photochemical efficiency and increased capability of thermal dissipation. A linear correlation was found between nonphotochemical quenching and xanthophyll cycle components. Thus, the change in carotenoids demonstrated an important role in photoprotection, the xanthophyll cycle was the main way of dissipation of excess energy under stress conditions in alfalfa.

Increased [CO2] Causes Changes in Physiological and Genetic Responses in C4 Crops: A Brief Review.

Climate change not only worries government representatives and organizations, but also attracts the attention of the scientific community in different contexts. In agriculture specifically, the cultivation and productivity of crops such as sugarcane, maize, and sorghum are influenced by several environmental factors. The effects of high atmospheric concentration of carbon dioxide ([CO2]) have been the subject of research investigating the growth and development of C4 plants. Therefore, this brief review presents some of the physiological and genetic changes in economically important C4 plants following exposure periods of increased [CO2] levels. In the short term, with high [CO2], C4 plants change photosynthetic metabolism and carbohydrate production. The photosynthetic apparatus is initially improved, and some responses, such as stomatal conductance and transpiration rate, are normally maintained throughout the exposure. Protein-encoding genes related to photosynthesis, such as the enzyme phosphoenolpyruvate carboxylase, to sucrose accumulation and to biomass growth and are differentially regulated by [CO2] increase and can variably participate owing to the C4 species and/or other internal and external factors interfering in plant development. Despite the consensus among some studies, mainly on physiological changes, further studies are still necessary to identify the molecular mechanisms modulated under this condition. In addition, considering future scenarios, the combined effects of high environmental and [CO2] stresses need to be investigated so that the responses of maize, sugarcane, and sorghum are better understood.

Serum phospholipidomics reveals altered lipid profile and promising biomarkers in multiple sclerosis

Multiple sclerosis is a neurodegenerative disease causing disability in young adults. Alterations in metabolism and lipid profile have been associated with this disease. Several studies have reported changes in the metabolism of arachidonic acid and the profile of fatty acids, ceramides, phospholipids and lipid peroxidation products. Nevertheless, the understanding of the modulation of circulating lipids at the molecular level in multiple sclerosis remains unclear. In the present study, we sought to assess the existence of a distinctive lipid signature of multiple sclerosis using an untargeted lipidomics approach. It also aimed to assess the differences in lipid profile between disease status (relapse and remission). For this, we used hydrophilic interaction liquid chromatography coupled with mass spectrometry for phospholipidomic profiling of serum samples from patients with multiple sclerosis. Our results demonstrated that multiple sclerosis has a phospholipidomic signature different from that of healthy controls, especially the PE, PC, LPE, ether-linked PE and ether-linked PC species. Plasmalogen PC and PE species, which are natural endogenous antioxidants, as well as PC and PE polyunsaturated fatty acid esterified species showed significantly lower levels in patients with multiple sclerosis and patients in both remission and relapse of multiple sclerosis. Our results show for the first time that the serum phospholipidome of multiple sclerosis is significantly different from that of healthy controls and that few phospholipids, with the lowest p-value, such as PC(34:3), PC(36:6), PE(40:10) and PC(38:1) may be suitable as biomarkers for clinical applications in multiple sclerosis.

Installation of C4 photosynthetic pathway enzymes in rice using a single construct.

SummaryIntroduction of a C4 photosynthetic mechanism into C3 crops offers an opportunity to improve photosynthetic efficiency, biomass and yield in addition to potentially improving nitrogen and water use efficiency. To create a two‐cell metabolic prototype for an NADP‐malic enzyme type C4 rice, we transformed Oryza sativa spp. japonica cultivar Kitaake with a single construct containing the coding regions of carbonic anhydrase, phosphoenolpyruvate (PEP) carboxylase, NADP‐malate dehydrogenase, pyruvate orthophosphate dikinase and NADP‐malic enzyme from Zea mays, driven by cell‐preferential promoters. Gene expression, protein accumulation and enzyme activity were confirmed for all five transgenes, and intercellular localization of proteins was analysed. 13CO2 labelling demonstrated a 10‐fold increase in flux though PEP carboxylase, exceeding the increase in measured in vitro enzyme activity, and estimated to be about 2% of the maize photosynthetic flux. Flux from malate via pyruvate to PEP remained low, commensurate with the low NADP‐malic enzyme activity observed in the transgenic lines. Physiological perturbations were minor and RNA sequencing revealed no substantive effects of transgene expression on other endogenous rice transcripts associated with photosynthesis. These results provide promise that, with enhanced levels of the C4 proteins introduced thus far, a functional C4 pathway is achievable in rice.

A Partial C4 Photosynthetic Biochemical Pathway in Rice

Introduction of a C4 photosynthetic pathway into C3 rice (Oryza sativa) requires installation of a biochemical pump that concentrates CO2 at the site of carboxylation in modified bundle sheath cells. To investigate the feasibility of this, we generated a quadruple line that simultaneously accumulates four of the core C4 photosynthetic enzymes from the NADP-malic enzyme subtype, phosphoenolpyruvate carboxylase (ZmPEPC), NADP-malate dehydrogenase (ZmNADP-MDH), NADP-malic enzyme (ZmNADP-ME), and pyruvate phosphate dikinase (ZmPPDK). This led to enhanced enzyme activity and mild phenotypic perturbations but was largely neutral in its effects on photosynthetic rate. Measurements of the flux of 13CO2 through photosynthetic metabolism revealed a significant increase in the incorporation of 13C into malate, consistent with increased fixation of 13CO2 via PEP carboxylase in lines expressing the maize PEPC enzyme. However, there was no significant differences in labeling of 3-phosphoglycerate (3PGA) indicating that there was no carbon flux through NADP-ME into the Calvin-Benson cycle. There was also no significant difference in labeling of phosphoenolpyruvate (PEP) indicating that there was no carbon flux through PPDK. Crossing the quadruple line with a line with reduced glycine decarboxylase H-protein (OsGDCH) abundance led to a photosynthetic phenotype characteristic of the reduced OsGDCH line and higher labeling of malate, aspartate and citrate than in the quintuple line. There was evidence of 13C labeling of aspartate indicating 13CO2 fixation into oxaloacetate by PEPC and conversion to aspartate by the endogenous aspartate aminotransferase activity. While Kranz anatomy or other anatomical modifications have not yet been installed in these plants to enable a fully functional C4 cycle, these results demonstrate for the first-time a partial flux through the carboxylation phase of NADP-ME C4 metabolism in transgenic rice containing two of the key metabolic steps in the C4 pathway.

Elevated temperature and waterlogging decrease cottonseed quality by altering the accumulation and distribution of carbohydrates, oil and protein.

Soil waterlogging and high-temperature events have occurred simultaneously in recent years in the Yangtze River basin cotton belt region of China, negatively affecting the development and quality of cottonseed. This study investigated the effects of the combination of elevated temperature (ET) (34.1/29.0°C) and waterlogging (3 or 6 days) on the accumulation and distribution of oil, protein and carbohydrates in cottonseed during flowering and boll development. The results showed that ET resulted in greater decreases in cottonseed biomass under waterlogging than under control conditions. The combination of waterlogging and ET significantly limited the accumulation of carbohydrates and oil contents. However, ET promoted protein accumulation and compensated for the negative effects of 3-day waterlogging on the final protein content. The combined ET and 6-day waterlogging significantly decreased the final contents of oil and protein by limiting carbon flux and NADPH supply because of the decreased activities of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) and glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49). The PEPC activity was correlated more with protein content than oil content. In addition, simultaneous exposure to waterlogging and ET resulted in lower unsaturated fatty acid/saturated fatty acid ratios and essential amino acid/non-essential amino acid ratios than did exposure to the individual factors alone. These findings could provide the theoretical support for the prospective assessment of effects of high temperature and waterlogging stresses on cotton production under climate change, and they can help to develop effective techniques in cotton cultivation.

Gene Dispensability in Escherichia coli Grown in Thirty Different Carbon Environments.

Central metabolism is a topic that has been studied for decades, and yet, this process is still not fully understood in Escherichia coli, perhaps the most amenable and well-studied model organism in biology. To further our understanding, we used a high-throughput method to measure the growth kinetics of each of 3,796 E. coli single-gene deletion mutants in 30 different carbon sources. In total, there were 342 genes (9.01%) encompassing a breadth of biological functions that showed a growth phenotype on at least 1 carbon source, demonstrating that carbon metabolism is closely linked to a large number of processes in the cell. We identified 74 genes that showed low growth in 90% of conditions, defining a set of genes which are essential in nutrient-limited media, regardless of the carbon source. The data are compiled into a Web application, Carbon Phenotype Explorer (CarPE), to facilitate easy visualization of growth curves for each mutant strain in each carbon source. Our experimental data matched closely with the predictions from the EcoCyc metabolic model which uses flux balance analysis to predict growth phenotypes. From our comparisons to the model, we found that, unexpectedly, phosphoenolpyruvate carboxylase (ppc) was required for robust growth in most carbon sources other than most trichloroacetic acid (TCA) cycle intermediates. We also identified 51 poorly annotated genes that showed a low growth phenotype in at least 1 carbon source, which allowed us to form hypotheses about the functions of these genes. From this list, we further characterized the ydhC gene and demonstrated its role in adenosine efflux.IMPORTANCE While there has been much study of bacterial gene dispensability, there is a lack of comprehensive genome-scale examinations of the impact of gene deletion on growth in different carbon sources. In this context, a lot can be learned from such experiments in the model microbe Escherichia coli where much is already understood and there are existing tools for the investigation of carbon metabolism and physiology (1). Gene deletion studies have practical potential in the field of antibiotic drug discovery where there is emerging interest in bacterial central metabolism as a target for new antibiotics (2). Furthermore, some carbon utilization pathways have been shown to be critical for initiating and maintaining infection for certain pathogens and sites of infection (3-5). Here, with the use of high-throughput solid medium phenotyping methods, we have generated kinetic growth measurements for 3,796 genes under 30 different carbon source conditions. This data set provides a foundation for research that will improve our understanding of genes with unknown function, aid in predicting potential antibiotic targets, validate and advance metabolic models, and help to develop our understanding of E. coli metabolism.

Differential expression in leaves of Saccharum genotypes contrasting in biomass production provides evidence of genes involved in carbon partitioning

BackgroundThe development of biomass crops aims to meet industrial yield demands, in order to optimize profitability and sustainability. Achieving these goals in an energy crop like sugarcane relies on breeding for sucrose accumulation, fiber content and stalk number. To expand the understanding of the biological pathways related to these traits, we evaluated gene expression of two groups of genotypes contrasting in biomass composition.ResultsFirst visible dewlap leaves were collected from 12 genotypes, six per group, to perform RNA-Seq. We found a high number of differentially expressed genes, showing how hybridization in a complex polyploid system caused extensive modifications in genome functioning. We found evidence that differences in transposition and defense related genes may arise due to the complex nature of the polyploid Saccharum genomes. Genotypes within both biomass groups showed substantial variability in genes involved in photosynthesis. However, most genes coding for photosystem components or those coding for phosphoenolpyruvate carboxylases (PEPCs) were upregulated in the high biomass group. Sucrose synthase (SuSy) coding genes were upregulated in the low biomass group, showing that this enzyme class can be involved with sucrose synthesis in leaves, similarly to sucrose phosphate synthase (SPS) and sucrose phosphate phosphatase (SPP). Genes in pathways related to biosynthesis of cell wall components and expansins coding genes showed low average expression levels and were mostly upregulated in the high biomass group.ConclusionsTogether, these results show differences in carbohydrate synthesis and carbon partitioning in the source tissue of distinct phenotypic groups. Our data from sugarcane leaves revealed how hybridization in a complex polyploid system resulted in noticeably different transcriptomic profiles between contrasting genotypes.

Anaplerotic Pathways in Halomonas elongata: The Role of the Sodium Gradient.

Salt tolerance in the γ-proteobacterium Halomonas elongata is linked to its ability to produce the compatible solute ectoine. The metabolism of ectoine production is of great interest since it can shed light on the biochemical basis of halotolerance as well as pave the way for the improvement of the biotechnological production of such compatible solute. Ectoine belongs to the biosynthetic family of aspartate-derived amino-acids. Aspartate is formed from oxaloacetate, thereby connecting ectoine production to the anaplerotic reactions that refill carbon into the tricarboxylic acid cycle (TCA cycle). This places a high demand on these reactions and creates the need to regulate them not only in response to growth but also in response to extracellular salt concentration. In this work, we combine modeling and experiments to analyze how these different needs shape the anaplerotic reactions in H. elongata. First, the stoichiometric and thermodynamic factors that condition the flux distributions are analyzed, then the optimal patterns of operation for oxaloacetate production are calculated. Finally, the phenotype of two deletion mutants lacking potentially relevant anaplerotic enzymes: phosphoenolpyruvate carboxylase (Ppc) and oxaloacetate decarboxylase (Oad) are experimentally characterized. The results show that the anaplerotic reactions in H. elongata are indeed subject to evolutionary pressures that differ from those faced by other gram-negative bacteria. Ectoine producing halophiles must meet a higher metabolic demand for oxaloacetate and the reliance of many marine bacteria on the Entner-Doudoroff pathway compromises the anaplerotic efficiency of Ppc, which is usually one of the main enzymes fulfilling this role. The anaplerotic flux in H. elongata is contributed not only by Ppc but also by Oad, an enzyme that has not yet been shown to play this role in vivo. Ppc is necessary for H. elongata to grow normally at low salt concentrations but it is not required to achieve near maximal growth rates as long as there is a steep sodium gradient. On the other hand, the lack of Oad presents serious difficulties to grow at high salt concentrations. This points to a shared role of these two enzymes in guaranteeing the supply of oxaloacetate for biosynthetic reactions.

Validation of an Enzyme-Driven Model Explaining Photosynthetic Rate Responses to Limited Nitrogen in Crop Plants.

The limited availability of nitrogen (N) is a fundamental challenge for many crop plants. We have hypothesized that the relative crop photosynthetic rate (P) is exponentially constrained by certain plant-specific enzyme activities, such as ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (NADP-G3PDH), 3-phosphoglyceric acid (PGA) kinase, and chloroplast fructose-1,6-bisphosphatase (cpFBPase), in Triticum aestivum and Oryza sativa. We conducted a literature search to compile information from previous studies on C3 and C4 crop plants, to examine the photosynthetic rate responses to limited leaf [N] levels. We found that in Zea mays, NADP-malic enzyme (NADP-ME), PEP carboxykinase (PCK), and Rubisco activities were positively correlated with P. A positive correlation was also observed between both phosphoenolpyruvate carboxylase (PEPC) and Rubisco activity with leaf [N] in Sorghum bicolor. Key enzyme activities responded differently to P in C3 and C4 plants, suggesting that other factors, such as leaf [N] and the stage of leaf growth, also limited specific enzyme activities. The relationships followed the best fitting exponential relationships between key enzymes and the P rate in both C3 and C4 plants. It was found that C4 species absorbed less leaf [N] but had higher [N] assimilation rates (A rate) and higher maximum photosynthesis rates (Pmax), i.e., they were able to utilize and invest more [N] to sustain higher carbon gains. All C3 species studied herein had higher [N] storage (Nstore) and higher absorption of [N], when compared with the C4 species. Nstore was the main [N] source used for maintaining photosynthetic capacity and leaf expansion. Of the nine C3 species assessed, rice had the greatest Pmax, thereby absorbing more leaf [N]. Elevated CO2 (eCO2) was also found to reduce the leaf [N] and Pmax in rice but enhanced the leaf [N] and N use efficiency of photosynthesis in maize. We concluded that eCO2 affects [N] allocation, which directly or indirectly affects Pmax. These results highlight the need to further study these physiological and biochemical processes, to better predict how crops will respond to eCO2 concentrations and limited [N].

Molecular Cloning of Novel-Type Phosphoenolpyruvate Carboxylase Isoforms in Pitaya (Hylocereus undatus).

Phosphoenolpyruvate carboxylase (PEPC) is an important enzyme involved in the initial CO2 fixation of crassulacean acid metabolism (CAM) photosynthesis. To understand the cultivation characteristics of a CAM plant pitaya, it is necessary to clarify the characteristics of PEPC in this species. Here, we cloned three PEPC cDNAs in pitaya, HuPPC1, HuPPC2, and HuPPC3, which encode 942, 934, and 966 amino acid residues, respectively. Phylogenetic analysis indicated that these PEPC belonged to plant-type PEPC (PTPC), although HuPPC1 and HuPPC2 have no Ser-phosphorylation motif in N-terminal region, which is a crucial regulation site in PTPC and contributes to CAM periodicity. HuPPC1 and HuPPC2 phylogenetically unique to the Cactaceae family, whereas HuPPC3 was included in a CAM clade. Two isoforms were partially purified at the protein level and were assigned as HuPPC2 and HuPPC3 using MASCOT analysis. The most distinct difference in enzymatic properties between the two was sensitivity to malate and aspartate, both of which are allosteric inhibitors of PEPC. With 2 mM malate, HuPPC3 was inhibited to 10% of the initial activity, whereas HuPPC2 activity was maintained at 70%. Aspartate inhibited HuPPC3 activity by approximately 50% at 5 mM; however, such inhibition was not observed for HuPPC2 at 10 mM. These results suggest that HuPPC3 corresponds to a general CAM-related PEPC, whereas HuPPC1 and HuPPC2 are related to carbon and/or nitrogen metabolism, with a characteristic regulation mechanism similar to those of Cactaceae plants.

Morpho-physiological and molecular responses of two Libyan bread wheat cultivars to plant growth regulators under salt stress

To study the effects of salt stress and plant growth regulators (kinetin, gibberellic acid, potassium) on growth, yield, glycine betaine content, phosphoenolpyruvate carboxylase (PEPC) and ribulose biphosphate carboxylase (RBC) gene expression of two Libyan bread wheat varieties, a factorial design of greenhouse experiment with three replications was conducted. Results revealed that salt stress significantly reduced plant growth and productivity of both varieties. Moreover, the addition of kinetin + potassium and gibberellic acid + potassium had improved the performance of the morpho-metric parameters of both genotypes under salt stress; but the performance was more effective for kinetin treatment than for gibberellic acid. At the biochemical level, the results showed that salt stress increased glycine betaine contents in both varieties with different proportions. This increase is more elevated in the presence of kinetin + potassium than the treatment with gibberellic acid+ potassium, which showed an almost similar result as in only salt stress. At the molecular level, the effects of salt stress and plant growth regulators on the PEPC and RBC gene expression showed that the increase was significantly higher for kinetin, gibberellic acid, and salt stress when compared to the control.
 Highlights - Salt stress reduced plant growth and productivity of bread wheat varieties. - Growth regulator improved the performance of the morphometric parameters. - The performance was more effective for kinetin treatment than for gibberellic acid. - Kinetin improved the glycine betaine gene expression more than gibberellic acid. - Kinetin increased significantly the phosphoenolpyruvate carboxylase and ribulose biphosphate carboxylase gene expression.

Differences in Physiological Responses of Two Oat (Avena nuda L.) Lines to Sodic-Alkalinity in the Vegetative Stage

Sodic-alkalinity is a more seriously limiting factor in agricultural productivity than salinity. Oat (Avena nuda) is a salt-tolerant crop species and is therefore useful in studying the physiological responses of cereals to alkalinity. We evaluated the differential effects of sodic-alkalinity on two naked oat lines, Caoyou1 and Yanke1. Seedlings of the two lines were exposed to 50 mM alkaline salt mixture of NaHCO3 and Na2CO3 (18:1 molar ratio; pH 8.5) for 2 weeks in a soil environment. Sodic-alkalinity exposure led the assimilation of abundant Na+ at similar concentrations in the organs of both lines. However, Caoyou1 showed much stronger growth than Yanke1, exhibiting a higher dry weight, total leaf area, and shoot height under sodic-alkalinity. Further analysis showed that Caoyou1 was more sodic-alkalinity tolerance than Yanke1. This was firstly because of differences in the oxidative stress defense mechanisms in leaves of the two lines. Antioxidant enzyme activities were either slightly elevated (catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GP), glutathione reductase (GR)) or unaltered (superoxide dismutase (SOD)) in Caoyou1 leaves, but some enzyme (SOD, GPOX, GR) activities were significantly reduced in Yanke1. AnAPX1 transcript levels significantly increased in Caoyou1 under sodic-alkalinity conditions compared with Yanke1, indicating its better antioxidant capacity. Secondly, the related parameters of Mg2+ concentration, phosphoenolpyruvate carboxylase (PEPC) activity, and AnPEPC transcript levels in the leaves showed significantly higher values in Caoyou1 compared with Yanke1. This demonstrated the effective utilization by Caoyou1 of accumulated HCO3− in the irreversible reaction from phosphoenolpyruvate to oxaloacetate to produce inorganic phosphorus, which was elevated in Caoyou1 leaves under alkalinity stress. Overall, the results demonstrated that the greater sodic-alkalinity tolerance of Caoyou1 is the result of: (1) maintained antioxidant enzyme activities; and (2) a higher capacity for the phosphoenolpyruvate to oxaloacetate reactions, as shown by the higher PEPC activity, Mg2+ concentration, and total phosphorus concentration in its leaves, despite the lower soil pH.

Repeated Negative Urine Trypsinogen-2 Dipstick Test Rules Out Diagnosis of Post-ERCP Pancreatitis

A dipstick test for urine trypsinogen-2 has been used in the diagnosis of acute pancreatitis, but there are only a few studies exploring the effectiveness of this test for early diagnose of post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP). The authors explore if the rapid point-of-care urine trypsinogen-2 dipstick test can replace assay of amylase in diagnosing PEP. For this prospective study, from Helsinki University Hospital 400 ERCP patients were enrolled in whom the authors analyzed plasma amylase or pancreas-specific amylase, bilirubin, and urine trypsinogen-2, and urine trypsinogen-2 with dipstick before, 4 and 24 hours after ERCP. PEP developed in 15 (3.8%) patients. Urine trypsinogen-2 concentrations were significantly higher in PEP than in non-PEP patients 24 hours after ERCP (P=0.001, Mann-Whitney U test) but not 4 hours after ERCP (P=0.094). When combined with abdominal pain symptoms at 4 hours the dipstick test had a sensitivity of 60%, a specificity of 99%, a positive predictive value of 64%, and a negative predictive value 98%. At 24 hours, sensitivity was 100%, specificity 98%, positive predictive value 71%, and negative predictive value 100%. A positive dipstick seems to identify PEP cases and a negative test excludes PEP with high accuracy.

Temporal trends and mortality of post-ERCP pancreatitis in the United States: a nationwide analysis.

BACKGROUND : Post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP) is the most common and most serious complication of ERCP. Our aim was to estimate the nationwide incidence, temporal trends, and mortality of PEP in the United States and to establish risk factors associated with PEP development. METHODS : This was a retrospective cohort study analyzing Nationwide Inpatient Sample data from 2011 to 2017 using International Classification of Diseases codes. The primary outcomes were trends in PEP incidence and predictors of PEP development. Secondary outcomes were in-hospital mortality, length of hospital stay, and admission to the intensive care unit. RESULTS : Of 1 222 467 adult patients who underwent inpatient ERCP during the study period, 55 225 (4.5 %) developed PEP. The hospital admission rate of PEP increased by 15.3 %, from 7735 in 2011 to 8920 in 2017 (odds ratio [OR] 1.23, 95 % confidence interval [CI] 1.04 - 1.46; P = 0.02). The overall rate of mortality increased from 2.8 % of PEP cases in 2011 to 4.4 % in 2017 (OR 1.62, 95 %CI 1.10 - 2.38; P = 0.01). Multiple patient-related (alcohol use, cocaine use, obesity, chronic kidney disease, heart failure), procedure-related (therapeutic ERCP, sphincterotomy, pancreatic duct stent placement, sphincter of Oddi dysfunction), and hospital-related (teaching hospitals, hospitals located in the West and Midwest) factors that impact the occurrence of PEP were identified. CONCLUSIONS : Our study showed rising hospital admission and mortality rates associated with PEP in the United States. This calls for a greater recognition of this life-threatening complication and amelioration of its risk factors, whenever possible.

Nitrate and ammonium differ in their impact on δ13C of plant metabolites and respired CO2 from tobacco leaves

ABSTRACT The carbon isotopic composition (δ13C) of foliage is often used as proxy for plant performance. However, the effect of vs. supply on δ13C of leaf metabolites and respired CO2 is largely unknown. We supplied tobacco plants with a gradient of to concentration ratios and determined gas exchange variables, concentrations and δ13C of tricarboxylic acid (TCA) cycle intermediates, δ13C of dark-respired CO2, and activities of key enzymes nitrate reductase, malic enzyme and phosphoenolpyruvate carboxylase. Net assimilation rate, dry biomass and concentrations of organic acids and starch decreased along the gradient. In contrast, respiration rates, concentrations of intercellular CO2, soluble sugars and amino acids increased. As decreased, activities of all measured enzymes decreased. δ13C of CO2 and organic acids closely co-varied and were more positive under supply, suggesting organic acids as potential substrates for respiration. Together with estimates of intra-molecular 13C enrichment in malate, we conclude that a change in the anaplerotic reaction of the TCA cycle possibly contributes to 13C enrichment in organic acids and respired CO2 under supply. Thus, the effect of vs. on δ13C is highly relevant, particularly if δ13C of leaf metabolites or respiration is used as proxy for plant performance.

SCARECROW gene function is required for photosynthetic development in maize.

C4 photosynthesis in grasses relies on a specialized leaf anatomy. In maize, this “Kranz” leaf anatomy is patterned in part by the duplicated SCARECROW (SCR) genes ZmSCR1 and ZmSCR1h. Here we show that in addition to patterning defects, chlorophyll content and levels of transcripts encoding Golden2‐like regulators of chloroplast development are significantly lower in Zmscr1; Zmscr1h mutants than in wild‐type. These perturbations are not associated with changes in chloroplast number, size, or ultrastructure. However, the maximum rates of carboxylation by ribulose bisphosphate carboxylase/oxygenase (RuBisCO, V cmax) and phosphoenolpyruvate carboxylase (PEPC, V pmax) are both reduced, leading to perturbed plant growth. The CO2 compensation point and 13C‰ of Zmscr1;Zmscr1h plants are both normal, indicating that a canonical C4 cycle is operating, albeit at reduced overall capacity. Taken together, our results reveal that the maize SCR genes, either directly or indirectly, play a role in photosynthetic development.Significance statementSCARECROW (SCR) is one of the best studied plant developmental regulators, however, its role in downstream plant physiology is less well‐understood. Here, we have demonstrated that SCR is required to establish and/or maintain photosynthetic capacity in maize leaves.

Tomato fruit as a model for tissue-specific gene silencing in crop plants

Use of CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR-associated 9)-mediated genome editing has proliferated for use in numerous plant species to modify gene function and expression, usually in the context of either transient or stably inherited genetic alternations. While extremely useful in many applications, modification of some loci yields outcomes detrimental to further experimental evaluation or viability of the target organism. Expression of Cas9 under a promoter conferring gene knockouts in a tissue-specific subset of genomes has been demonstrated in insect and animal models, and recently in Arabidopsis. We developed an in planta GFP (green fluorescent protein) assay system to demonstrate fruit-specific gene editing in tomato using a phosphoenolpyruvate carboxylase 2 gene promoter. We then targeted a SET-domain containing polycomb protein, SlEZ2, previously shown to yield pleiotropic phenotypes when targeted via 35S-driven RNA interference and we were able to characterize fruit phenotypes absent additional developmental perturbations. Tissue-specific gene editing will have applications in assessing function of essential genes otherwise difficult to study via germline modifications and will provide routes to edited genomes in tissues that could not otherwise be recovered when their germline modification perturbs their normal development.