Glycerophosphocholine Research Articles

Choline Alphoscerat (Gliatilin) in the pathogenetic therapy of Alzheimer’s disease

Alzheimer’s disease (Alzheimer’s type dementia) (BA) is a primary progressive neurodegenerative disease of the brain with characteristic clinical and pathological features, individual characteristics of the course and severity of symptoms, as well as multiple converging etiopathogenetic mechanisms. Te most successfully developed at the present time are compensatory (replacement) therapy aimed at overcoming the neurotransmitter defciency in various neuronal systems, which are more or less affected by the development of Alzheimer’s disease. Choline alfoscerat (Gliatilin) is a cholinomimetic of central action with a predominant effect on the central nervous system. In the process of metabolism, the molecule of Gliatilin (choline alphoscerate) undergoes cleavage into choline (40.5%) and glycerophosphate. Te resulting choline stimulates the synthesis of acetylcholine, which improves the transmission of nerve impulses in cholinergic neurons, and glycerophosphate stimulates the appearance of phosphatidylcholine, which restores the phospholipid composition of neuronal membranes, improving their plasticity. Glitatylin (choline alfoscerate) has a stimulating effect on cognitive functions and improves memory and attention, leads to the correction of psycho-emotional disorders. Glitatilin (choline alphosceratus) positively increases in cerebral blood flow, accelerates metabolism and activates structure brain reticular formation. Te effect on glia is expressed in suppressing the activation of inflammation, reducing intracellular edema and reducing microvascular permeability. Choline alfostserat (Gliatilin) can be recommended for use in patients with asthma as monotherapy, and about complex treatment.

Lipoic acid-derived cross-linked liposomes for reduction-responsive delivery of anticancer drug

Liposomes have emerged as a fascinating nanocarriers for the delivery of cancer therapeutics. However, their efficacy for cancer therapy is reduced partially because of the serum-instability and incomplete drug release. In this study, a novel disulfide cross-linked liposomes (CLs) assembled from dimeric lipoic acid-derived glycerophosphorylcholine (di-LA-PC) conjugate was developed. The conjugate was synthesized by a facial esterification of lipoic acid (LA) and glycerophosphorylcholine (GPC) and characterized by MS, 1H NMR and 13C NMR. Featuring the enhanced serum-stability and intracellular drug release determined by in vitro stability and GSH-responsive behavior, CLs prepared with dried thin film technique following 10 % dithiothreitol (DTT) cross-linking can attain effective delivery of anticancer candidates. Notably, CLs stably encapsulated doxorubicin (Dox) in their vesicular structures and showed a remarkable thiol-sensitive release of payload upon cellular uptake by cancer cells, compared to that of uncross-linked liposomes (uCLs) or Doxil-like liposome (DLLs). The cell viability and apoptosis of Dox-loaded CLs worked the pronounced cytotoxic effects to MCF-7 cells with an IC50 value of 10.8 μg Dox equiv./mL comparable to free Dox and 2.8-fold higher than DLLs. More importantly, it is demonstrated that the nanoscale characteristics of Dox-loaded CLs could prevent the proliferation of adriamycin-resistant MCF-7/ADR cell line, highlighting their potential in reversal of drug resistance. Furthermore, the preliminary in vivo test (n = 3) showed that disulfide cross-linked liposomal formulation of Dox (Dox-CLs) improved the therapeutic efficacy compared to free Dox and DLLs in a human breast carcinoma xenograft mouse model. Therefore, the current thiol-responsive cross-linked liposome may provide a robust drug delivery platform for cancer therapy.

Neurometabolite changes in patients with complex regional pain syndrome using magnetic resonance spectroscopy: a pilot study.

The aim of this study was to investigate distinct neurometabolites in the right and left thalamus and insula of patients with complex regional pain syndrome (CRPS) compared with healthy controls using proton magnetic resonance spectroscopy. Levels of N-acetylaspartate (NAA), N-acetylaspartylglutamate (NAAG), myo-inositol (ml), glutamine (Gln), glycerophosphocholine (GPC), glutathione (GSH), and alanine (Ala) relative to total creatine (tCr) levels, including creatine and phosphocreatine, were determined in the right and left thalamus and insula in 12 patients with CRPS compared with 11 healthy controls using magnetic resonance spectroscopy. Levels of NAAG/tCr and Ala/tCr were higher in patients with CRPS than in controls in the left thalamus. NAAG/tCr, ml/tCr, and Gln/tCr levels were higher but NAA/tCr levels were lower in the right insula of patients with CRPS compared with controls. There were negative correlations between GSH/tCr and pain score (McGill Pain Questionnaire) in the left thalamus. These findings are paramount to understand and determine all aspects of the complex pathophysiological mechanisms that underlie CRPS, including involvement of the central and parasympathetic nervous systems as well as oxidative stress and antioxidants. Thus, the distinct metabolites presented herein may be essential to understand a strong diagnostic and prognostic potential for CRPS and to develop effective medical treatments.

The effect of repeated isoflurane exposure on serine synthesis pathway during the developmental period in Caenorhabditis elegans

BackgroundSerine synthetic pathway plays an essential role in the development and function of the nervous system. This study investigated whether the serine synthetic pathway was affected by repeated volatile anesthetic exposure using C. elegans and its relationship with anesthesia-induced neurotoxicity. MethodsSynchronized worms were divided into two groups: the control and isoflurane groups. Worms in the isoflurane group were exposed to isoflurane for 1 h at each larval stage. The chemotaxis index was evaluated when they reached the young adult-stage in both groups. Also, RNA was extracted from the young adult-worms, and the expressions of C31C9.2, F26H9.5, and Y62E10 A.13 were evaluated using real-time polymerase chain reaction in both groups. At the same time, the l-serine level was measured. After phosphoserine phosphatase inhibitor – glycerophosphorylcholine (GPC) – and l-serine were treated, the change of chemotaxis index was determined. ResultsIn young adult worms exposed to isoflurane, the genetic expressions of C31C9.2, F26H9.5, and Y62E10 A.13 were decreased, and a significant decrease was shown in Y62E10 A.13. The serine level in worms was also lower in the isoflurane group than in the control group (5.13 ± 1.44 vs. 7.65 ± 0.81 pM, n = 5 in each group, p = 0.009). Exposure to GPC reduced the chemotaxis index to a similar degree as repeated isoflurane exposure (52.9% in GPC group vs 58.7% in the isoflurane group). The chemotaxis index (61.1%) was not decreased by repeated isoflurane anesthesia in GPC-treated worms. In this condition, the l-serine level was low similarly in both groups (5.22 ± 1.19 vs. 4.90 ± 1.36 pM, n = 5 in each group, p = 0.702). When l-serine was supplied to C. elegans, the deteriorated chemotaxis index by isoflurane exposure recovered (78.1% in the control group vs. 75.5% in the isoflurane group, p = 0.465). ConclusionSerine synthetic pathway was negatively affected in C. elegans by repeated isoflurane exposure. Y62E10 A.13, which corresponds to phosphoserine phosphatase, was mostly influenced, followed by low l-serine level. Supplementation with l-serine could restore the chemotaxis index.

The glycerophosphocholine acyltransferase Gpc1 is part of a phosphatidylcholine (PC)-remodeling pathway that alters PC species in yeast

Phospholipase B-mediated hydrolysis of phosphatidylcholine (PC) results in the formation of free fatty acids and glycerophosphocholine (GPC) in the yeast Saccharomyces cerevisiae GPC can be reacylated by the glycerophosphocholine acyltransferase Gpc1, which produces lysophosphatidylcholine (LPC), and LPC can be converted to PC by the lysophospholipid acyltransferase Ale1. Here, we further characterized the regulation and function of this distinct PC deacylation/reacylation pathway in yeast. Through in vitro and in vivo experiments, we show that Gpc1 and Ale1 are the major cellular GPC and LPC acyltransferases, respectively. Importantly, we report that Gpc1 activity affects the PC species profile. Loss of Gpc1 decreased the levels of monounsaturated PC species and increased those of diunsaturated PC species, whereas Gpc1 overexpression had the opposite effects. Of note, Gpc1 loss did not significantly affect phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine profiles. Our results indicate that Gpc1 is involved in postsynthetic PC remodeling that produces more saturated PC species. qRT-PCR analyses revealed that GPC1 mRNA abundance is regulated coordinately with PC biosynthetic pathways. Inositol availability, which regulates several phospholipid biosynthetic genes, down-regulated GPC1 expression at the mRNA and protein levels and, as expected, decreased levels of monounsaturated PC species. Finally, loss of GPC1 decreased stationary phase viability in inositol-free medium. These results indicate that Gpc1 is part of a postsynthetic PC deacylation/reacylation remodeling pathway (PC-DRP) that alters the PC species profile, is regulated in coordination with other major lipid biosynthetic pathways, and affects yeast growth.

Analysis of compensatory metabolic pathway for taurine deficiency in mouse heart

Taurine is one of the most abundant free amino acids in heart, and its treatment is beneficial against chronic heart failure. While knocking out taurine transporter (TauT) in mice caused a taurine-deficient cardiomyopathy, cardiac output is normal in the young TauTKO mouse, suggesting activation of homeostatic mechanisms that compensate for detrimental effects caused by taurine depletion. In the present study, we examined the integrated analysis of transcriptome and metabolome of TauTKO mice to determine the compensatory homeostatic pathway. We identified increases in several organic osmolytes, including betaine, carnitine, glycerophosphocholine (GPC) and amino acids, in the heart of TauTKO mice. Transcriptome analysis revealed that several genes of the SLC transporter family are increased in TauTKO mice, such as amino acid transporter (Slc38a2), while the established transporters for betaine and carnitine are not altered by taurine deficiency. The integrated analysis revealed a significant increase in the genes involved in Glycerophospholipid metabolism in which GPC biosynthesis is involved. In conclusion, we identified genetic/metabolic compensatory mechanisms involved in the control of the metabolome profile in taurine-deficient cardiomyopathy.

GDE5 inhibition accumulates intracellular glycerophosphocholine and suppresses adipogenesis at a mitotic clonal expansion stage.

Mammalian glycerophosphodiesterases (GDEs) were recently shown to be involved in multiple cellular signaling pathways. This study showed that decreased GDE5 expression results in accumulation of intracellular glycerophosphocholine (GPC), showing that GDE5 is actively involved in GPC/choline metabolism in 3T3-L1 adipocytes. Using 3T3-L1 adipocytes, we further studied the biological significance of GPC/choline metabolism during adipocyte differentiation. Inhibition of GDE5 suppressed the formation of lipid droplets, which is accompanied by the decreased expression of adipocyte differentiation markers. We further showed that the decreased GDE5 expression suppressed mitotic clonal expansion (MCE) of preadipocytes. Decreased expression of CTP: phosphocholine cytidylyltransferase (CCTβ), a rate-limiting enzyme for phosphatidylcholine (PC) synthesis, is similarly able to inhibit MCE and PC synthesis; however, the decreased GDE5 expression resulted in accumulation of intracellular GPC but did not affect PC synthesis. Furthermore, we showed that mRNAs of proteoglycans and transporters for organic osmolytes are significantly upregulated and that intracellular amino acids and urea levels are altered in response to GDE5 inhibition. Finally, we showed that reduction of GDE5 expression increased lactate dehydrogenase release from preadipocytes. These observations indicate that decreased GDE5 expression can suppress adipocyte differentiation not through the PC pathway but possibly by intracellular GPC accumulation. These results provide insight into the roles of mammalian GDEs and their dependence upon osmotic regulation by altering intracellular GPC levels.

The role of acetylcholine neurotransmission in the pharmacotherapy of cognitive dysfunction

Microangiopathy and deficit in cholinergic system innervating cerebral cortex and controlling the plasticity of neuronal structuresplay an important role in the formation of cognitive disorders. The main approaches to the therapy of dementia and organic lesions followed by cognitive disorder are based on the hypothesis of replenishment of acetylcholine. Choline alfoscerate takes a special place among the precursors of choline due to its proven effectiveness in the treatment of ischemic and neurodegenerative lesions of the central nervous system. The article presents the materials of experimental and clinical studies of the drug choline alfostserat (Gliatilin).

Absolute Quantification of Phosphor-Containing Metabolites in the Liver Using 31 P MRSI and Hepatic Lipid Volume Correction at 7T Suggests No Dependence on Body Mass Index or Age.

BackgroundHepatic disorders are often associated with changes in the concentration of phosphorus‐31 (31P) metabolites. Absolute quantification offers a way to assess those metabolites directly but introduces obstacles, especially at higher field strengths (B0 ≥ 7T).PurposeTo introduce a feasible method for in vivo absolute quantification of hepatic 31P metabolites and assess its clinical value by probing differences related to volunteers' age and body mass index (BMI).Study TypeProspective cohort.Subjects/PhantomsFour healthy volunteers included in the reproducibility study and 19 healthy subjects arranged into three subgroups according to BMI and age. Phantoms containing 31P solution for correction and validation.Field Strength/SequencePhase‐encoded 3D pulse‐acquire chemical shift imaging for 31P and single‐volume 1H spectroscopy to assess the hepatocellular lipid content at 7T.AssessmentA phantom replacement method was used. Spectra located in the liver with sufficient signal‐to‐noise ratio and no contamination from muscle tissue, were used to calculate following metabolite concentrations: adenosine triphosphates (γ‐ and α‐ATP); glycerophosphocholine (GPC); glycerophosphoethanolamine (GPE); inorganic phosphate (Pi); phosphocholine (PC); phosphoethanolamine (PE); uridine diphosphate‐glucose (UDPG); nicotinamide adenine dinucleotide‐phosphate (NADH); and phosphatidylcholine (PtdC). Correction for hepatic lipid volume fraction (HLVF) was performed.Statistical TestsDifferences assessed by analysis of variance with Bonferroni correction for multiple comparison and with a Student's t‐test when appropriate.ResultsThe concentrations for the young lean group corrected for HLVF were 2.56 ± 0.10 mM for γ‐ATP (mean ± standard deviation), α‐ATP: 2.42 ± 0.15 mM, GPC: 3.31 ± 0.27 mM, GPE: 3.38 ± 0.87 mM, Pi: 1.42 ± 0.20 mM, PC: 1.47 ± 0.24 mM, PE: 1.61 ± 0.20 mM, UDPG: 0.74 ± 0.17 mM, NADH: 1.21 ± 0.38 mM, and PtdC: 0.43 ± 0.10 mM. Differences found in ATP levels between lean and overweight volunteers vanished after HLVF correction.Data ConclusionExploiting the excellent spectral resolution at 7T and using the phantom replacement method, we were able to quantify up to 10 31P‐containing hepatic metabolites. The combination of 31P magnetic resonance spectroscopy imaging data acquisition and HLVF correction was not able to show a possible dependence of 31P metabolite concentrations on BMI or age, in the small healthy population used in this study. Level of Evidence: 2 Technical Efficacy: Stage 1J. Magn. Reson. Imaging 2019;49:597–607.

Visceral fat-related systemic inflammation and the adolescent brain: a mediating role of circulating glycerophosphocholines.

Life-long maintenance of brain health is important for the prevention of cognitive impairment in older age. Low-grade peripheral inflammation associated with excess visceral fat (VF) may influence brain structure and function. Here we examined (i) if this type of inflammation is associated with altered white-matter (WM) microstructure and lower cognitive functioning in adolescents, and (ii) if recently identified circulating glycerophosphocholines (GPCs) can index this type of inflammation and associated variations in WM microstructure and cognitive functioning. We studied a community-based sample of 872 adolescents (12-18 years, 48% males) in whom we assessed VF and WM microstructure with magnetic resonance imaging, processing speed with cognitive testing, serum C-reactive protein (CRP, a common marker of peripheral inflammation) with a high-sensitivity assay, and serum levels of a panel of 64 GPCs with advanced mass spectrometry. VF was associated with CRP, and CRP in turn was associated with "altered" WM microstructure and lower processing speed (all p < 0.003). Further, "altered" WM microstructure was associated with lower processing speed (p < 0.0001). Of all 64 tested GPCs, 4 were associated with both VF and CRP (at Bonferroni corrected p < 0.0004). One of them, PC16:0/2:0, was also associated with WM microstructure (p < 0.0001) and processing speed (p = 0.0003), and mediated the directed associations between VF and both WM microstructure (p < 0.0001) and processing speed (p = 0.02). As a mediator, PC16:0/2:0 explained 21% of shared variance between VF and WM microstructure, and 22% of shared variance between VF and processing speed. Similar associations were observed in an auxiliary study of 80 middle-aged adults. Our results show that VF-related peripheral inflammation is associated with "altered" WM microstructure and lower cognitive functioning already in adolescents, and a specific circulating GPC may be a new molecule indexing this VF-related peripheral inflammation and its influences on brain structure and function.

Synthesis of CLA‐Rich Lysophosphatidylcholine by Immobilized MAS1‐H108A‐Catalyzed Esterification: Effects of the Parameters and Monitoring of the Reaction Process

Conjugated linoleic acid (CLA)‐rich lysophosphatidylcholine (LPC) with many proven beneficial effects is successfully synthesized by an immobilized mutant lipase (MAS1‐H108A)‐catalyzed esterification of glycerophosphorylcholine (GPC) and CLA‐rich FAs. Under the optimized conditions (temperature of 55 °C, substrate molar ratio of CLA‐rich FAs to GPC of 40:1, and enzyme loading of 300 U g−1), LPC content as high as 89.10 mol.% is achieved, which is exceptionally higher than any ever‐reported value. By monitoring the esterification process, it is found that sn1‐LPC is easy to synthesize and is the predominant product, while sn2‐LPC and phosphatidylcholine (PC) are difficult to synthesize and are with lower content in the final product. The formed sn2‐LPC during esterification may mainly attribute to the acyl migration of sn1‐LPC and the ratio of sn1‐LPC to sn2‐LPC finally plateaus at approximately 7. Besides, our results also demonstrate that sn2‐LPC is the main template for the synthesis of PC. Finally, a complete reaction scheme for the synthesis of LPC by immobilized MAS1‐H108A‐catalyzed esterification of GPC and fatty acids is mapped out. Overall, the highest LPC content can be obtained by immobilized MAS1‐H108A‐catalyzed esterification and there is the first study for systematical studying the reaction process of CLA‐rich LPC synthesis.Practical Applications: Previous studies have demonstrated that Novozym 435 is the most suitable catalyst for the synthesis of CLA‐rich LPC. However, the LPC content obtained by Novozym 435 is only 70 mol.% and the reaction process for the synthesis of CLA‐rich LPC has not been studied systematically, which restricts the commercial production of CLA‐rich LPC in some extent. Therefore, further exploration of alternative enzyme resources for highly efficient synthesis of CLA‐rich LPC and trying to clarify the reaction process of CLA‐rich LPC synthesis are of significant importance for the future commercially industrial production of CLA‐rich LPC.A complete reaction scheme for the synthesis of LPC using GPC and FAs as substrates by immobilized MAS1‐H108A‐catalyzed esterification. CLA‐rich FAs are used as model FAs in this study. Reaction parameters, such as temperature, CLA‐rich FAs/GPC ratio, and enzyme loading are selected to study their effects on the esterification. Concurrently, the reaction process is monitored. LPC content as high as 89.10 mol.% is obtained, which is the highest reported value thus far for the synthesis of CLA‐rich LPC. Finally, the reaction scheme for the synthesis of LPC by immobilized MAS1‐H108A‐catalyzed esterification is mapped out.

METABOLIC THERAPY IN PATIENTS WITH ISCHEMIC STROKE

The article shows the world experience of metabolic therapy use in the treatment of ischemic stroke. The issue still remains prominent. The reasonability of prescribing metabolic drugs is not completely clear, its effectiveness has not been fully proved, despite numerous studies which show only trends. The article presents an overview of the most popular drugs of different pharmacological groups with a metabolic effect which affect different parts of the ischemic cascade. Ethylmethylhydroxypyridine succinate and cytoflavin have predominantly antihypoxic effect, improve functional outcome and neurological functions, and normalize overall well-being and adaptation. Cerebrolysin is a complex of low molecular weight biologically active peptides derived from the pig’s brain. It has a multimodal effect on the brain, helps to reduce the volume of cerebral infarction, restores neurologic functions and improves the functional outcome. Cortexin is a mixture of cattle brain polypeptides, also has a complex action that provides the most complete reversion of neurological deficit, improves cognitive functions and the functional outcome, reduces the level of paroxysmal convulsive readiness and improves bioelectric activity of the brain. Citicoline is a precursor of cell membrane key ultrastructures, contributes to significant reduction in the volume of cortical brain damage, improves cholinergic transmission, which results in better clinical outcome, even despite the questionable impact on the neurological status. Choline Alfoscerate is a precursor of choline, and the use of the drug significantly limits the growth of the cerebral infarction area starting from the first day of therapy, leads to reversion of neurological symptoms and achievement of rehabilitation goals. Actovegin is deproteinized derivative of calf blood, activates metabolism in tissues, improves trophism and stimulates regeneration. In a large study, it was shown that Actovegin improved cognitive function in patients who had experienced a stroke,. The drug does not significantly improve the neurological status of patients after a stroke, but it reduces the risk of the stroke development in the next 10 years. Thus, we analyzed mechanisms of medical substances action and data of experimental and clinical studies, including ones after thrombolytic therapy and with inclusion of drugs for primary and secondary prevention of ischemic stroke. The reasonability and effectiveness of prescribing a combination of drugs of different pharmacological groups affecting brain metabolism remains controversial, since the excessive drug treatment may have complications. The safety of metabolic therapy is in doubt, and some of authors views presented confirm the need for additional large independent studies.

P300 latency changes in patients with mild cognitive impairment after taking choline alphoscerate; A preliminary study.

Choline alphoscerate in clinical studies improved cognitive dysfunction in dementia, but it did not show any clear clinical benefit on mild cognitive impairment (MCI). There is limited evidence of neuropsychological markers in showing the effects of cholinergic precursors in MCI. Object of this preliminary study is to evaluate the change of the P300 latency as a biomarker for cognitive function after taking choline alphoscerate in patients with MCI. Event related evoked potential study were done in baseline (n = 27) and 3 months after taking choline alphoscerate (n = 17). When compared to our previous reported control database, the difference of the P300 latencies between MCI and control group at baseline was statistically significant (P < 0.01). Although Follow-up P300 latencies after taking choline alphoscerate did not show the significant change, the tendency of shortened P300 latencies was identified. Even though there are some limitations, choline alphoscerate could improve the electrophysiological markers in MCI patients. To identify the effect of cholinergic precursor in MCI and the usefulness of electrophysiological biomarkers, well-designed further study is needed.

Headache in elderly patients with chronic cerebral ischemia: outpatient diagnosis and treatment

Chronic cerebral ischemia (CCI) is one of the most common diagnoses in middle-aged and elderly patients in the practice of an outpatient neurologist. Unfortunately, the diagnosis of CCI in these patients is often established only on the basis of complaints of headache, dizziness, instability during walking, and lower mood. At the same time, other diseases that cause these symptoms are not diagnosed, patients do not receive treatment, which considerably worsens quality of life and leads to anxiety and depression. A variety of diseases, such as headache, peripheral vestibular vertigo, depression, Alzheimer's disease, and Parkinson's syndrome, are frequently hidden under the diagnosis of CCI. The leading neurological syndrome in CCI is cognitive impairment that can be both moderate and reach the level of dementia. Approximately 40% of patients with chronic cerebrovascular disease complain of headache that is usually caused by mixed primary headache. The management tactics for a CCI patient suffering from headache is aimed at treating primary headache, modifying vascular risk factors, and managing cognitive impairment. The paper discusses the use of choline alphoscerate in patients diagnosed with CCI.

The Glycerophosphocholine Acyltransferase, Gpc1, Impacts PC Remodeling and Stationary Phase Cell Viability in Saccharomyces cerevisiae

Phosphatidylcholine (PC) is the most abundant glycerophospholipid found in most eukaryotic membranes. Changes in its synthesis, turnover and remodeling play a crucial role in maintaining membrane homeostasis. PC species vary depending upon the acyl chains attached at the sn‐1 and sn‐2 position of the glycerol backbone. The fatty acid components of yeast PC are relatively simple, consisting of mostly C16:0, C16:1, C18:0 and C18:1 combined to produce PC species in which either one or both acyl chains is monounsaturated. Thus, the four major PC species found in yeast consist predominantly of the following chain combinations: two C16:1 to produce 32:2 PC, C16:0 and C16:1 to produce 32:1PC, C16:1 and C18:1 to produce 34:2PC, C16:0 and C18:1 or 18:0 and C16;1 to produce 34:1PC. Acyl chain composition is important as it affects the physical properties of membranes such as thickness, curvature and fluidity. We have recently reported on the characterization of glycerophosphocholine acyltransferase, Gpc1. Gpc1 acylates glycerophosphocholine (GPC), the product of the complete deacylation of PC, to produce lysophosphatidylcholine (LysoPC). LysoPC can then be converted to PC by the lysophospholipd acyltransferase, Ale1. In order to investigate the role of Gpc1 in PC remodeling, we performed PC species analyses on strains lacking or overexpressing GPC1. Our data indicates that the loss of GPC1 results in a decrease in PC species with a single unsaturation (32:1 and 34:1) and an increase in PC species with two unsaturations (32:2 and 34:2). Conversely, the overexpression of GPC1 results in an increase in singly unsaturated PC species (32:1 and 34:1). Expression analyses indicate that the GPC1 is upregulated upon attenuation of PC and sphingolipid biosynthetic pathways. Finally, preliminary phenotypic studies indicate that while loss of GPC1 does not affect log phase doubling time, it does result in a decrease in long‐term cell survival. Overall, we provide evidence that the remodeling of PC to more saturated species by Gpc1 is regulated in coordination with other lipid biosynthetic pathways and impacts stationary phase cell viability.Support or Funding InformationWe acknowledge support from National Institute of Health Grant NIH R15 GM104876 and Duquesne University.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effect of Mono-, Di-, and Trihydric Alcohols on Lipase-Catalyzed Alcoholysis of Phosphatidylcholine in Hexane

In this study, alcoholysis of phosphatidylcholine (PC) catalyzed by two lipases (Novozym 435 and Lipozyme RM IM) was performed in n-hexane to evaluate the effects of mono-, di-, and trihydric alcohols (ethanol, 1-butanol, 1,2-ethanediol, 1,2-propanediol, and glycerol) on the alcoholysis reactions. In the reactions, as PC was converted into lysophosphatidylcholine (LPC), LPC was simultaneously alcoholyzed and converted into glycerophosphorylcholine (GPC). The alcohols affected the alcoholysis of both PC and LPC. When alcoholysis was catalyzed by Novozym 435, the reaction efficiencies of the selected alcohols in the alcoholysis of PC followed the order 1,2-propanediol ≈ ethylene glycol> 1-butanol > ethanol > glycerol, and the reaction efficiencies of these alcohols in the alcoholysis of LPC were 1,2-propanediol > ethylene glycol> 1-butanol ≈ glycerol > ethanol. For alcoholysis catalyzed by Lipozyme RM IM, the reaction efficiencies of alcohols in alcoholysis of PC followed the order ethylene glycol ≈1,2-propanediol > glycerol > ethanol > 1-butanol, and the reaction efficiencies in the alcoholysis of LPC were ethylene glycol > glycerol > 1,2-propanediol > ethanol > 1-butanol. In general, in the lipase-catalyzed alcoholysis of PC, reaction efficiencies with dihydric alcohols are higher than those with mono- and trihydric alcohols.Supported by National Natural Science Foundation of China (No. 31772003), and Basic Research Funds in Henan University of Technology (No. 2017RCJH07).

Effect of Electroacupuncture at "Neiguan"(PC 6) on Serum and Myocardial Metabolites in Rats with Myocardial Ischemia Reperfusion Injury Based on Nuclear Magnetic Resonance Spectroscopy

We have repeatedly demonstrated that electroacupuncture (EA) of "Neiguan"(PC 6) can improve myocardial ischemia in rats. The present study was designed to investigate the metabolomic profile of peripheral blood se-rum and myocardium involving EA-induced improvement of myocardial ischemia-reperfusion injury (MIRI) in rats by using nuclear magnetic resonance spectroscopy. Thirty male SD rats were equally randomized into blank control, model and EA groups. Rats of the control group were only banded for 20 min, once a day for 7 days. The MIRI model was established by occlusion of the anterior descending branch of the left coronary artery for 40 min, followed by reperfusion for 60 min, and rats of the model group were banded as those in the control group. EA (10 Hz/50 Hz, 1 mA) was applied to bilateral PC 6 for 20 min, once daily for 7 days. The blood samples and left ventricular myocardial tissues were collected for assaying the profiles of differential metabolites using 1H nuclear magnetic resonance (1H NMR) spectroscopy and multivariate statistical analysis such as the principal components analysis (PCA), partial least squares-discriminant analysis (PLS-DA) and orthogonal PLS-DA (O-PLS-DA) with SIMCA-P software 12.0. A total of 19 differential metabolites (17 down-regulated, 2 up-regulated) in the serum and 14 differential metabolites (13 down-regulated and 1 up-regulated) in the ischemic left myocardium were identified after MIRI. Of the 19 serum differential metabolites, amino acids (leucine, isoleucine, valine,alanine, lysine, glycine, glutamine), 3-hydroxy butyric acid (3-HB), lactic acid, acetate, N-acetyl glycoprotein (NAc), acetone, acetoacetate, succinate, polyunsaturated fatty acids (PUFA), creatine, glycerophosphocholine (GPC) were down-regulated; while low density lipoprotein (LDL), LDL/very low density lipoprotein(LDL/VLDL)and glucose obviously up-regulated. Of the 14 myocardial differential metabolites, amino acids (alanine, lysine, glutamate, glutamine, aspartate, taurine, glycine, threonine), GPC, creatine, lactic acid, adenosine monophosphate (AMP), nicotinamide adenine dinucleotide (NAD＋) were significantly decreased, and glucose was up-regulated. Following EA treatment, most of the decreased serum differential metabolites except acetone, acetoacetate and PUFA, and the increased serum LDL, LDL/VLDL and glucose recovered, basically close to the control level; and the decreased myocardial creatine, GPC and NAD＋ were also apparently up-regulated and the increased myocardial glucose was down-regulated. But, myocardial threonine and AMP still presented a decreasing state. Although the pattern of myocardial differential metabolites of the EA group had a trend to be close to the control group, the significant difference still existed, while the metabolic pattern of serum metabolites in the EA group was close to that of the control group. EA stimulation of PC 6 can regulate serum or/and myocardial metabolites as amino acids, carbohydrates, lipids, etc. in MIRI rats, of which both serum and myocardial creatine, GPC and glucose may be jointly confer a favorable potential for EA-induced improvement of MIRI.

P.1.014 - Investigating the effect of central nervous system drugs on leukocyte elastase activity in vitro

Participation of immune reactions in the pathogenesis of mental diseases is widely studied. A variety of molecules are involved in the development of inflammatory reactions, one of the analyzed biochemical markers of inflammation is the activity of leukocyte elastase (LE), an enzyme secreted by neutrophils. Due to proteolytic activity towards elastin and collagen, LE is able to disrupt the integrity of the vessels and increase permeability of the blood-brain barrier in case of various brain diseases [1]. The effect of drugs used to treat mental illness (or lack thereof) on the inflammatory component of the pathogenesis of mental diseases seems to be poorly understood. Aim This study was а comparative assessment of the effect of psychotropic (chlorpromazine, imipramine) and nootropic (cerebrolysin, ceretone) drugs on LE activity in an in vitro experiment. Material and Methods LE activity was measured in a pool of blood plasma of healthy people with solutions of drugs: chlorpromazine; imipramine; choline alphoscerate; cerebrolysin in different concentrations, including therapeutic ones. Enzymatic activity of LE was determined by spectrophotometric method. Statistical comparison of multiple groups was carried out using ANOVA and Newman-Keuls test, values of p Results А dose-dependent inhibitory effect of chlorpromazine was revealed. In the range of concentrations from 1.25 μg/ml to 40 μg/ml, including therapeutic one of 5-10 μg/ml, LE activity was significantly lower compared to control (p Different concentrations (0.00375 mg/ml - 3.75 mg/ml) of choline alfoscerate had no influence on LE activity (p = 0.33). Cerebrolysin at concentrations from 0.043 mg/ml to 2.15 mg/ml also had no influence on LE activity, however, at concentration 4.3 mg/ml, which corresponds to a double maximum dose, cerebrolysin had a potentiating effect on LE, whose activity significantly increased by 17.5% compared to control (p Conclusion Thus, it has been shown that psychotropic drugs such as chlorpromazine and imipramine have an inhibitory effect on LE activity in an in vitro experiment, while nootropic cerebrolysin can activate LE. The obtained data indicate that these drugs, used in the treatment of mental illness, can influence inflammatory response, which is important in the pathogenesis of mental disorders.

Liposomes of dimeric artesunate phospholipid: A combination of dimerization and self-assembly to combat malaria

Artemisinin and its derivatives are highly effective drugs in the treatment of P. falciparum malaria. However, their clinical applications face challenges because of short half-life, poor bioavailability and growing drug resistance. In this article, novel dimeric artesunate phospholipid (Di-ART-GPC) based liposomes were developed by combination of dimerization and self-assembly to address these shortcomings. Firstly, Di-ART-GPC conjugate was synthesized by a facile esterification of artesunate (ART) and glycerophosphorylcholine (GPC) and confirmed by MS, 1H NMR and 13C NMR. The conjugate was then assembled to form liposomes without excipient by thin film hydration method. The assembled Di-ART-GPC liposomes have typical multilamellar vesicle structure with bilayer morphology as determined by transmission electron microscopy (TEM) and cryogenic electron microscopy (cryo-EM). Moreover, the liposomes displayed an average hydrodynamic diameter of 190 nm and negative zeta potential at −20.35 mV as determined by Zetasizer. The loading capacity of ART was calculated approximately 77.6% by weight with this liposomal formulation after a simple calculation. In vitro drug release and degradation results showed that the Di-ART-GPC liposomes were stable in neutral physiological conditions but effectively degraded to release parent ART in simulated weakly acidic microenvironment. In vivo pharmacokinetics study revealed that Di-ART-GPC liposomes and conjugate have longer retention half-life in bloodstream. Importantly, Di-ART-GPC liposomes (IC50 0.39 nM) and the conjugate (IC50 1.90 nM) demonstrated excellent in vitro antiplasmodial activities without causing hemolysis of erythrocytes, which were superior to free ART (IC50 5.17 nM) and conventional ART-loaded liposomes (IC50 3.13 nM). Furthermore, the assembled liposomes resulted in enhanced parasites killing in P. berghei-infected mice in vivo with delayed recrudescence and improved survivability, compared to free ART administration. Based on these encouraging results, Di-ART-GPC liposomal formulation could be a replacement to parent ART in clinical malarial therapy after thorough investigation.

Modeling the effects of Irgarol 1051 on coral using lipidomic methodology for environmental monitoring and assessment

Coral is commonly selected as a bioindicator of detecting a variety of adverse factors such as photosystem II herbicide Irgarol 1051, through measuring pan-type biomarkers. To improve the effectiveness of biomonitoring, omic technologies have recently been applied to model the systemic changes in an organism. Membrane lipids create a dynamic cell structure based on the physiological state, which offers a distinct lipid profile to specifically detect environmental threats and assess the associated health risk. To demonstrate the potential of a lipidomic methodology for biomonitoring, the glycerophosphocholine (GPC) profiles of the coral Seriatopora caliendrum were observed during 3 days of Irgarol (0.1–2.0 μg/L) exposure. The lipid profile variations were modeled based on the Irgarol dose and the coral photoinhibition levels to develop an excellent quantitative model. The predominant changes correlated with the photoinhibition, decreasing the lyso-GPCs and GPCs with lower unsaturated chains and increasing GPCs with highly polyunsaturated chains, can be related to the consequence of blocking the photosynthetic electron flow based on the associated physiological roles. Other dose-specific lipid changes led to the partial exchange of PC(O-16:0/20:5) for PC(16,0/20:5) as a first-line response to counteract the membrane opening caused by Irgarol. Increased levels of the GPCs with 20:4 or 22:6 chains, which can promote mitochondrial functionality, confirmed an elevated respiration level in the coral exposed to Irgarol levels of >0.5 μg/L. Notably, plasmanylcholines with 20:4 or 22:6 chains and phosphatidylcholines with 22:6 or 22:5 chains, which can alter their membrane material properties to mitigate organelle pre-swelling and swelling in different ways, formed in the coral exposed to the 0.5 and 2.0 μg/L Irgarol levels. Such coral adaptations further predict the health risks associated with altered physiological conditions. In this study, the lipidomic methodology is demonstrated as a potential tool for environmental monitoring and assessment.