Characterization of Molecular Properties and Expression of Gene GmPLMT and Its Effects on the Production of Lipid Metabolites in Soybean and Arabidopsis thaliana

Phospholipid N-methyltransferase (PLMT) plays an important role in the synthesis of phosphatidylcholine (PtdCho). The aim of this study was to characterize the molecular properties of GmPLMT and the expression of soybean GmPLMT and its effects on the production of lipid metabolites. Results showed that GmPLMT composed of mainly α-helix was a hydrophobic and transmembrane protein. In soybean leaves, GmPLMT was highly expressed during seedling and flowering stages. In transgenic Arabidopsis thaliana, the highest and lowest expression levels of GmPLMT were detected at flowering and maturity stages, respectively. The total phospholipid contents in soybean grains were decreased from 7.2% (35 days after flowering) to 4.8% (55 days after flowering) and then increased to 7.0% (75 days after flowering). The contents of PtdCho showed a similar pattern to that of total phospholipids. In transgenic A. thaliana seeds, the contents of total phospholipids and PtdCho were significantly increased. Significantly positive correlations were revealed between expression of GmPLMT and contents of both PtdCho and crude fats, and between the contents of PtdCho and both linoleic acid and linolenic acid, suggesting that increased expression of GmPLMT improved the production of lipid metabolites. This study provided solid experimental evidence for further improvement of soybean quality based on GmPLMT in the molecular breeding of soybeans.

Soil microbial respiration, biomass carbon and community composition data induced by substrate quality from an agricultural grassland

The data presented here are related to the research article entitled ‘Effects ofsubstrate quality on carbon partitioning and microbial community composition in soil from an agricultural grassland’ [1]. Data illustrate cumulative CO2 efflux, microbial biomass C (Cmic), priming effect expressed as priming index (PI) and total phospholipid fatty acid (PLFA) profiles. The data were measured during four soil laboratory incubations using a silty clay loam soil under permanent grassland from May until August 2015. The soil was treated with carbohydrates of different complexity (glucose, glucose-6-phosphate (G6P) or cellulose) alone or in conjunction with livestock slurry amended or non-amended with a biological additive. Our data may be of great significance for further studies on microbial respiration and biosynthesis, and microbial community structure following slurry application to soil, alongside the potential beneficial effects of the addition of slurry amended with biological additives.

Molecular Patterns of Extreme and Persistent Cholesterol Efflux Capacity

Objective: Cholesterol efflux capacity (CEC), the ability of extracellular acceptors to pick-up cholesterol from macrophages, is a clinically relevant cardiovascular biomarker. CEC is inversely associated with incident atherosclerotic cardiovascular disease events. However, CEC is only modestly associated with HDL-C (high-density lipoprotein cholesterol) levels, which may explain the failure of HDL-C raising therapies to improve atherosclerotic cardiovascular disease outcomes. Determinants of variation in CEC are not well understood. Thus, we sought to establish whether extreme high and low CEC is a robust persistent phenotype and to characterize associations with cholesterol, protein, and phospholipids across the particle size distribution. Approach and Results: CEC was previously measured in 2924 participants enrolled in the Dallas Heart Study, a multi-ethnic population-based study from 2000 to 2002. We prospectively recruited those who were below the 10th and above 90th percentile of CEC. Our study revealed that extreme low and high CEC are persistent, robust phenotypes after 15 years of follow-up. Using size exclusion chromatography, CEC to fractionated plasma depleted of apolipoprotein B (fraction-specific CEC) demonstrated significant differences in CEC patterns between persistent high and low efflux groups. Fraction-specific CEC was correlated with fraction-specific total phospholipid but not apolipoprotein A-I, cholesterol, or total protein. These correlations varied across the size distribution and differed among persistent high versus low efflux groups. Conclusions: Extreme high and low CEC are persistent and robust phenotypes. CEC patterns in fractionated plasma reveal marked variation across the size distribution. Future studies are warranted to determine specific molecular species linked to CEC in a size-specific manner.

Linkage of microbial living communities and residues to soil organic carbon accumulation along a forest restoration gradient in southern China

Background Forest restoration has been considered an effective method to increase soil organic carbon (SOC), whereas it remains unclear whether long-term forest restoration will continuously increase SOC. Such large uncertainties may be mainly due to the limited knowledge on how soil microorganisms will contribute to SOC accumulation over time. Methods We simultaneously documented SOC, total phospholipid fatty acids (PLFAs), and amino sugars (AS) content across a forest restoration gradient with average stand ages of 14, 49, 70, and > 90 years in southern China. Results The SOC and AS continuously increased with stand age. The ratio of fungal PLFAs to bacterial PLFAs showed no change with stand age, while the ratio of fungal AS to bacterial AS significantly increased. The total microbial residue-carbon (AS-C) accounted for 0.95–1.66 % in SOC across all forest restoration stages, with significantly higher in fungal residue-C (0.68–1.19 %) than bacterial residue-C (0.05–0.11 %). Furthermore, the contribution of total AS-C to SOC was positively correlated with clay content at 0–10 cm soil layer but negatively related to clay content at 10–20 cm soil layer. Conclusions These findings highlight the significant contribution of AS-C to SOC accumulation along forest restoration stages, with divergent contributions from fungal residues and bacterial residues. Soil clay content with stand age significantly affects the divergent contributions of AS-C to SOC at two different soil layers.

Reduced Organic Carbon Content during the Evolvement of Calcareous Soils in Karst Region

Understanding the changes in soil organic carbon (SOC) storage is important for accurately predicting ecosystem C sequestration and/or potential C losses, but the relevant information, especially for the evolvement of calcareous soil is limited in karst regions. Three calcareous soils with different evolvement intensities were sampled from an evergreen broadleaved forest in the subtropical region of southwest of China to investigate the changes in different SOC fractions and microbial communities. The results showed that: (1) The contents of SOC, dissolved organic carbon (DOC), mineral protected organic carbon (MOC), and recalcitrant organic carbon (ROC) significantly decreased with increasing evolvement intensity of calcareous soil, but pH and the chemical composition of SOC, including Alkyl C, O-alkyl C, Aromatic C, and Carbonyl C, did not significantly change, suggesting that various SOC fractions synergistically decrease with the evolvement of calcareous soil. (2) The evolvement of calcareous soil had a substantial negative effect on total phospholipid fatty acids (PLFA), bacteria (i.e., Gram positive bacteria and Gram negative bacteria), fungi, and actinomycetes, but did not affect the ratio of fungi to bacteria. This result supported the conclusion that various SOC fractions were synchronously loss with the evolvement of calcareous soil. (3) Results from the multivariate statistical analysis showed a significant correlation between SOC fractions (including SOC, DOC, MOC, and ROC) and soil base cations, mainly calcium (Ca), iron (Fe), and aluminum (Al). This strengthens the fact that SOC stability largely depends on the complex relationship between organic matter and mineral composition in soil. Taken together, the reduction of SOC during the evolvement of soil in the karst areas accords with some mechanisms of previous studies (e.g., microbial composition and soil geochemistry), and also has its own unique characteristics (e.g., the relative contribution of carbons to chemical shift regions of CPMAS 13C-NMR spectra and F:B ratio).

Postmortem Analyses in a Patient With Succinic Semialdehyde Dehydrogenase Deficiency (SSADHD): II. Histological, Lipid, and Gene Expression Outcomes in Regional Brain Tissue

This study has extended previous metabolic measures in postmortem tissues (frontal and parietal lobes, pons, cerebellum, hippocampus, and cerebral cortex) obtained from a 37-year-old male patient with succinic semialdehyde dehydrogenase deficiency (SSADHD) who expired from SUDEP (sudden unexplained death in epilepsy). Histopathologic characterization of fixed cortex and hippocampus revealed mild to moderate astrogliosis, especially in white matter. Analysis of total phospholipid mass in all sections of the patient revealed a 61% increase in cortex and 51% decrease in hippocampus as compared to (n = 2-4) approximately age-matched controls. Examination of mass and molar composition of major phospholipid classes showed decreases in phospholipids enriched in myelin, such as phosphatidylserine, sphingomyelin, and ethanolamine plasmalogen. Evaluation of gene expression (RT2 Profiler PCR Arrays, GABA, glutamate; Qiagen) revealed dysregulation in 14/15 GABAA receptor subunits in cerebellum, parietal, and frontal lobes with the most significant downregulation in ∊, θ, ρ1, and ρ2 subunits (7.7-9.9-fold). GABAB receptor subunits were largely unaffected, as were ionotropic glutamate receptors. The metabotropic glutamate receptor 6 was consistently downregulated (maximum 5.9-fold) as was the neurotransmitter transporter (GABA), member 13 (maximum 7.3-fold). For other genes, consistent dysregulation was seen for interleukin 1β (maximum downregulation 9.9-fold) and synuclein α (maximal upregulation 6.5-fold). Our data provide unique insight into SSADHD brain function, confirming astrogliosis and lipid abnormalities previously observed in the null mouse model while highlighting long-term effects on GABAergic/glutamatergic gene expression in this disorder.

Protective effects of pulmonary surfactant on decompression sickness in rats

Decompression sickness (DCS) is a systemic pathophysiological process featured by bubble load . Lung dysfunction plays a harmful effect on off-gassing, which contributes to bubble load and subsequent DCS occurrence. This study aimed to investigate the effects of pulmonary surfactant on DCS as it possessed multiple advantages on the lung. Rats were divided into three groups: the normal (n = 10), the surfactant (n = 36) and the saline (n = 36) group. Animals in surfactant or saline group were administered aerosol surfactant or saline 12 h before a stimulated diving, respectively. Signs of DCS were recorded and bubble load was detected. The contents of phospholipid and surfactant protein A (SPA), protein, IL-1 and IL-6 in bronchoalveolar lavage fluid (BALF) and lung wet/dry (W/D) ratio were determined. Serum levels of IL-6, ICAM-1, E-selectin, GSH and GSSG were detected. In surfactant treated rats, the morbidity and mortality of DCS markedly decreased (**P < 0.01, *P < 0.05, respectively). Survival time prolonged and the latency to DCS dramatically delayed (**P < 0.01). More importantly, bubble load markedly decreased (**P < 0.01). The increases of protein, IL-1 and IL-6 in BALF and lung W/D ratio were alleviated. Restoration of total phospholipid and SPA in BALF and ICAM-1 and E-selectin in serum were observed. The inflammation and oxidation were attenuated (# P < 0.01). In conclusion, pre-diving administrating exogenous surfactant by aerosolization is an efficient, simple and safe method for DCS prevention in rats.

137 Dietary Supplementations of Ethanolamine and Serine Enhance Docosahexaenoic Acid Deposition in Eggs of Laying Hens

Enrichment of docosahexaenoic-acid (DHA) into eggs of laying-hens may be limited by the availability phospholipids as a deposition sink. The present study was to determine if dietary supplementations of phospholipid-component molecules or synthesis-enhancers: choline, serine, and ethanolamine could elevate phospholipid and DHA enrichment in the eggs and tissues of hens. A total of 50-White-Leghorn-Shavers (42-wk old) were individually caged and divided into 5 groups (n = 10/group). The 5 groups of hens were fed the following diets for 3 wk: Diet-1 = a corn soybean-meal basal-diet, Diet-2 = Diet-1 + 4%-microalgae (Aurantiochytrium, Heliae, Gibert, AZ, 1.81 g-DHA/kg) + choline-chloride (26.3 g/kg diet, 60% purity, DSM-Co., Basel, Switzerland), Diet-3 = Diet-2 + 1.41% of L-serine (100% purity, Ajinomoto-Co., Inc., Kawasaki, Japan), Diet-4 = Diet-2 + 100 mg of ethanolamine/kg (99% purity, Sigma-Aldirch-Co., St Loius, MO), and Diet-5 = Diet-3 + 100 mg of ethanolamine/kg. At the end of study, eggs, liver, ovary, and adipose samples were collected from 6 hens/group for biochemical analysis. Data were analyzed by one-way ANOVA. Compared with Diet-1, Diet-2 enhanced (P &lt; 0.05) DHA concentrations in egg yolk and liver by 213 mg/egg and 2.98 mg/g tissue, respectively, but decreased (P &lt; 0.05) total phospholipid-concentrations of yolk and liver by 50%, and hepatic-mRNA levels of elongases-2/5 and desaturases-4/6 by 25–50%. Compared with Diet-2, Diet-5 enhanced (P &lt; 0.05) DHA (by 20%) and phospholipid (by 40%) concentrations in the egg yolk, and upregulated (P &lt; 0.05) lipid-metabolism genes involved in the citicoline (CDP, up-to-3-fold) and CDP-ethanolamine (up-to-2.5-fold) pathways in the liver and ovary-tissue. In comparison, Diets-3 and 4 resulted in only 3–11% higher (P &lt; 0.05) DHA-concentrations in the liver over Diet-2. In conclusion, feeding hens a high DHA and choline diet enriched DHA in the egg yolk and down-regulated lipogenesis-gene-expression in the tissues. Supplementing the diet with extra-serine and ethanolamine further-enhanced the DHA enrichment in the egg yolk and restored the high DHA-mediated changes in the gene-expression. (Supported in part by DOE-MAGIC-grant DE-EE0007091, USDA-grant 2019-69012-29905, and Cornell-University-Hatch-grants NYC-127419/127302).

Phosphoinositide-Dependent Signaling in Cancer: A Focus on Phospholipase C Isozymes

Phosphoinositides (PI) form just a minor portion of the total phospholipid content in cells but are significantly involved in cancer development and progression. In several cancer types, phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P3] and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] play significant roles in regulating survival, proliferation, invasion, and growth of cancer cells. Phosphoinositide-specific phospholipase C (PLC) catalyze the generation of the essential second messengers diacylglycerol (DAG) and inositol 1,4,5 trisphosphate (InsP3) by hydrolyzing PtdIns(4,5)P2. DAG and InsP3 regulate Protein Kinase C (PKC) activation and the release of calcium ions (Ca2+) into the cytosol, respectively. This event leads to the control of several important biological processes implicated in cancer. PLCs have been extensively studied in cancer but their regulatory roles in the oncogenic process are not fully understood. This review aims to provide up-to-date knowledge on the involvement of PLCs in cancer. We focus specifically on PLCβ, PLCγ, PLCδ, and PLCε isoforms due to the numerous evidence of their involvement in various cancer types.