Combination Of C16 Research Articles

Adjusting vascular permeability, leukocyte infiltration, and microglial cell activation to rescue dopaminergic neurons in rodent models of Parkinson\u2019s disease

Animal studies have indicated that increased blood-brain barrier (BBB) permeability and inflammatory cell infiltration are involved during the progression of Parkinson’s disease (PD). This study used C16, a peptide that competitively binds to integrin αvβ3 and inhibits inflammatory cell infiltration, as well as angiopoietin-1 (Ang-1), an endothelial growth factor crucial for blood vessel protection, to reduce inflammation and improve the central nervous system (CNS) microenvironment in murine models of PD. The combination of C16 and Ang-1 yielded better results compared to the individual drugs alone in terms of reducing dopaminergic neuronal apoptosis, ameliorating cognitive impairment, and electrophysiological dysfunction, attenuating inflammation in the CNS microenvironment, and improving the functional disability in PD mice or rats. These results suggest neuroprotective and anti-inflammatory properties of the C16 peptide plus Ang-1 in PD.

Identification, isolation, and heterologous expression of Sunflower wax synthase for the synthesis of tailored wax esters.

Wax esters (WE) are neutral lipids formed by condensation of fatty alcohol with fatty acyl-CoA by wax synthases. They serve as carbon and energy reserves and are potential substrates for various commercial applications. Sunflower (Helianthus annuus) an edible oil seed is a source of WE, however, the gene responsible for WE formation has hitherto remained unidentified. Using an in silico approach we identified, isolated putative Sunflower wax synthase (HaWS) gene and investigated it's potential for WE production in yeast. Heterologous expression of HaWS in Saccharomyces cerevisiae H1246 exhibited 57kDa protein which was confirmed by immunoblotting. Recombinant yeast expressing HaWS were fed with combinations of C16, C18 fatty alcohols with 16:0, 18:0 fatty acyl CoA's as potential substrates to validate WE formation in vivo. The yeast cells accumulated C-32 to C-36 WE. Our study reveals identification, isolation, and heterologous functional expression of WS gene from Sunflower for the first time. PRACTICAL APPLICATIONS: Wax synthases (WSs) are critical enzymes for wax ester (WE) biosynthesis. WEs are high value products having several industrial applications. WE serve as substrates for lubricants, food coatings, cosmetics, and pharmaceuticals. There is a demand for alternate renewable resource of WEs. In this study, we have successfully isolated a putative wax synthase gene from Sunflower and submitted its sequence data to the GenBank (Accession number MH460820). Conserved sequence search analysis showed presence of condensation superfamily motif‒HHXXXDG, critical for WE biosynthesis. Heterologous expression ofHaWSin yeast revealed synthesis of C-32 to C-36 WE. Our study demonstrates the efficacy ofHaWSto accumulate specific WE of desired lengths in yeast, and thus represents an alternate source of WE for commercial applications and for biotechnological production of tailored WE in eukaryotic expression systems.

P-187 - Erythrocyte membrane fatty acids as the potential biomarkers for detection of early-stage and progression of colorectal cancer

Introduction: Colorectal cancer (CRC) is the third most common cancer in the world. Clinical data show that 5-year survival rate of early-stage CRC postoperative patients is around 90%. However, most of CRC patients were diagnosed at advanced stage due to its asymptomatic and poor diagnostic techniques. Early screening is an effective way to reduce the morbidity and mortality. So, it is necessary to develop low-cost, less invasive, high-sensitivity, and high-specificity screening methods for early diagnosis of CRC and its progression. The study aims to assess erythrocyte membrane fatty acids (FA) as the potential biomarkers for detection of early-stage from healthy controls and progression of colorectal cancer (CRC). Methods: Erythrocyte membrane FA from 95 patients (56 + 8 years old) with colorectal adenocarcinoma and 28 healthy people (the control group) were measured using an accelerated solvent extraction and analyzed by GC/MS system triple quad Agilent 7000B (USA). Tumors were further classified into early stage (stage I or II, n = 44) and advanced stage (stage III or IV, n = 51) based on TNM classification. Results: Erythrocyte membrane levels of C14:0 (p < 0.03), C15:0 (p < 0.04), C17:0 (p < 0.02), the sum of saturated FA (p < 0.03), C16:1 (p < 0.04), the sum of monounsaturated FA (p < 0.01), C18:2 (p < 0.01), omega-6/omega-3 (p < 0,01) in early stage CRC patients were significantly decreased compared with healthy controls, whereas the levels of C20:0 (p < 0.01), C20:2, (p < 0.01), C20:3 (p < 0.001), C20:4 (p < 0.001), C22:4 (p < 0.001), C22:5 (p < 0.001), C22:6 (p < 0.001) and the sum of all unsaturated FA (p < 0,03) were significantly higher than those from the controls. CRC progression was accompanied by decrease in the content of saturated, monounsaturated as well as by increase polyunsaturated fatty acids (p < 0.001-0.05). Elevated levels of polyunsaturated fatty acids as structural components of membranes reflect a high level of their instability, which is probably associated with the process of tumor proliferation. In contrast, a decrease in the level of saturated fatty acids may be due to their consumption as an energy substrate for rapid metabolism which is necessary for tumor growth. As saturated FA do not require creation of a double bond within the beta-oxidation process it allows them for an accelerated energy production. The panel, containing the FA - C17:0, C16:1, C18:2, C20:4, C22:5, C22:6 - achieved an excellent diagnostic performance when comparing early stage patients with healthy controls with an AUC of 0.959, a sensitivity of 86.5%, and a specificity of 96.2%. A combination of C16:0, C18:1;t9, C18:3, C20:2, C20:4, C20:5, C22:5, C22:6 showed the best diagnostic ability when comparing the late stage CRC patients with early stages (AUC 0.871, sensitivity of 82.7%, specificity of 80.8%. Conclusion: Erythrocyte levels of the fatty acids should be considered as the promising biomarkers for detecting of early stages of CRC and the progression of the disease.

Combination Treatment of C16 Peptide and Angiopoietin-1 Alleviates Neuromyelitis Optica in an Experimental Model.

Neuromyelitis optica (NMO) is an autoimmune inflammatory demyelinating disease that mainly affects the spinal cord and optic nerve, causing blindness and paralysis in some individuals. Moreover, NMO may cause secondary complement-dependent cytotoxicity (CDC), leading to oligodendrocyte and neuronal damage. In this study, a rodent NMO model, showing typical NMO pathogenesis, was induced with NMO-IgG from patient serum and human complement. We then tested whether the combination of C16, an αvβ3 integrin-binding peptide, and angiopoietin-1 (Ang1), a member of the endothelial growth factor family, could alleviate NMO in the model. Our results demonstrated that this combination therapy significantly decreased disease severity, inflammatory cell infiltration, secondary demyelination, and axonal loss, thus reducing neural death. In conclusion, our study suggests a possible treatment that can relieve progressive blindness and paralysis in an animal model of NMO through improvement of the inflammatory milieu.

Altering the ratio of dietary palmitic, stearic, and oleic acids in diets with or without whole cottonseed affects nutrient digestibility, energy partitioning, and production responses of dairy cows

The objective of this study was to evaluate the effects of varying the ratio of dietary palmitic (C16:0), stearic (C18:0), and oleic (cis-9 C18:1) acids in basal diets containing soyhulls or whole cottonseed on nutrient digestibility, energy partitioning, and production response of lactating dairy cows. Twenty-four mid-lactation multiparous Holstein cows were used in a split-plot Latin square design. Cows were allocated to a main plot receiving either a basal diet with soyhulls (SH, n = 12) or a basal diet with whole cottonseed (CS, n = 12) that was fed throughout the experiment. Within each plot a 4 × 4 Latin square arrangement of treatments was used in 4 consecutive 21-d periods. Treatments were (1) control (CON; no supplemental fat), (2) high C16:0 supplement [PA; fatty acid (FA) supplement blend provided ∼80% C16:0], (3) C16:0 and C18:0 supplement (PA+SA; FA supplement blend provided ∼40% C16:0 + ∼40% C18:0), and (4) C16:0 and cis-9 C18:1 supplement (PA+OA; FA supplement blend provided ∼45% C16:0 + ∼35% cis-9 C18:1). Interactions between basal diets and FA treatments were observed for dry matter intake (DMI) and milk yield. Among the SH diets, PA and PA+SA increased DMI compared with CON and PA+OA treatments, whereas in the CS diets PA+OA decreased DMI compared with CON. The PA, PA+SA, and PA+OA treatments increased milk yield compared with CON in the SH diets. The CS diets increased milk fat yield compared with the SH diets due to the greater yield of de novo and preformed milk FA. The PA treatment increased milk fat yield compared with CON, PA+SA, and PA+OA due to the greater yield of mixed-source (16-carbon) milk FA. The PA treatment increased 3.5% fat-corrected milk compared with CON and tended to increase it compared with PA+SA and PA+OA. The CS diets increased body weight (BW) change compared with the SH diets. Additionally, PA+OA tended to increase BW change compared with CON and PA and increased it in comparison with PA+SA. The PA and PA+OA treatments increased dry matter and neutral detergent fiber digestibility compared with PA+SA and tended to increase them compared with CON. The PA+SA treatment reduced 16-carbon, 18-carbon, and total FA digestibility compared with the other treatments. The CS diets increased energy partitioning toward body reserves compared with the SH diets. The PA treatment increased energy partitioning toward milk compared with CON and PA+OA and tended to increase it compared with PA+SA. In contrast, PA+OA increased energy partitioned to body reserves compared with PA and PA+SA and tended to increase it compared with CON. In conclusion, milk yield responses to different combinations of FA were affected by the addition of whole cottonseed in the diet. Among the combinations of C16:0, C18:0, and cis-9 C18:1 evaluated, fat supplements with more C16:0 increased energy output in milk, whereas fat supplements with more cis-9 C18:1 increased energy storage in BW. The combination of C16:0 and C18:0 reduced nutrient digestibility, which most likely explains the lower performance observed compared with other treatments.

Serum Unsaturated Free Fatty Acids: A Potential Biomarker Panel for Early-Stage Detection of Colorectal Cancer.

Background: To screen biomarkers to differentiate early-stage colorectal cancer (CRC) from benign colorectal disease (BCD) and healthy controls.Materials & Methods: Quantitative and qualitative analysis of C16:1, C18:3, C18:2, C18:1, C20:4, and C22:6 in 185 healthy controls, 55 patients with BCD, and 139 patients with CRC was performed. Comparisons of their levels in between CRC patients, BCD patients, and healthy controls were performed using Mann-Whitney U test.Results: Serum levels of C16:1, C18:3, C18:2, C18:1, C20:4, and C22:6 in CRC patients were significantly decreased compared with healthy controls and BCD patients. A combination of C16:1, C18:2, C20:4, and C22:6 has excellent diagnostic performance to differentiate early-stage CRC patients from healthy controls plus BCD patients, with an AUC of 0.926, a sensitivity of 84.6%, and a specificity of 89.8%.Conclusions: Serum levels of C16:1, C18:2, C20:4, and C22:6 could be diagnostic indicators of early-stage CRC patients.

Abstract 3511: Targeted epigenetic reprogramming reverts tumor progression in triple-negative breast cancer models by the activation of retinoic acid receptor alpha

Abstract INTRODUCTION: Triple negative breast cancer (TNBC) is associated with aggressiveness, early recurrence and poor prognosis and no other therapeutic alternative besides chemotherapy. The silencing of genes associated with cell differentiation in cancer is required for tumor progression. In this process the epigenetic modifications induced by aberrant control of the chromatin remodeling machinery in transformed cells plays a key role. Therefore, finding a mechanism to restore the expression of these genes by reverting the abnormal epigenetic modifications is an important task in the fight against cancer. We showed previously that the selective interference of the Sin3's PAH2 domain with Sin3 interaction domain (SID) mimicking peptides restored expression of E-cadherin, estrogen receptor and RARa target genes such as RARb and CRBP1 leading to cell differentiation and anti-tumor effect in TNBC cells. Here we studied whether the re-expression of retinoic acid receptors (RARs) induced by the small molecule C16 (SID decoy) can make TNBC cells sensitive to selective retinoid therapies. EXPERIMENTAL APPROACH: Human and mouse cell models of TNBC as well as MMTV-Myc oncomice were used to test the effects of the SID decoys on the induction of RAR signaling, cell proliferation, induction of differentiation in 3D cultures, expansion of the cancer stem cell compartment, atypical ductal hyperplasia (ADH), ductal carcinola in situ (DCIS), tumor development and metastatic dissemination. RESULTS: Our data show that C16 induced a differential expression of RARs, ∼10 fold upregulation of RARa2, RARbeta1/2, ∼10 fold downregulation of RARg1 and ∼30% increased production of retinoic acid (RA). The combination of C16 plus RARa agonists, AM80 (tamibarotene) or AM580 induced 75% inhibition of cell proliferation in 2D cultures, induced immature acini in 3D and impaired cancer stem cell proliferation inhibiting tumorsphere formation (∼65%) in vitro. In vivo, the combination of C16 and AM580 reduced 90% tumor growth and metastasis. Interestingly, the treatment of MMTV-Myc oncomice with C16 alone or in combination with AM80 prevented mammary gland hyperplasia, DCIS and delayed tumor development (80% tumor free survival, Kaplan-Meier analysis). CONCLUSION: As a whole, the epigenetic reprogramming of the TNBC cells is promising for design of therapies based on the use of RARa agonists to induce differentiation of the tumor cells; giving these patients a chance of an alternative or complementary therapy to chemotherapy regimens. Citation Format: Nidhi Bansal, Almudena Bosch, Boris A. Leibovitch, Keely Pierzchalski, Zhou Ming-Ming, Maureen Kane, Samuel Waxman, Eduardo Farias. Targeted epigenetic reprogramming reverts tumor progression in triple-negative breast cancer models by the activation of retinoic acid receptor alpha. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3511. doi:10.1158/1538-7445.AM2015-3511

Serum Unsaturated Free Fatty Acids: A Potential Biomarker Panel for Differentiating Benign Thyroid Diseases from Thyroid Cancer

Background: Serum free fatty acids (FFAs) are correlated with pathological status, and change in serum FFA levels may be associated with thyroid diseases.Materials and Methods: In this study, 664 serum samples from 322 healthy controls, 129 patients with benign thyroid disease (BTD), and 213 patients with thyroid cancer (TC) were collected. Chip-based direct-infusion nanoelectrospray-mass spectrometry was performed to simultaneously quantify six serum FFAs (i.e., C16:1, C18:1, C18:2, C18:3, C20:4, and C22:6.), with the excellent correlation coefficients of > 0.99 and relative standard deviation of <18% for all analysts. The Mann-Whitney U test was used to compare the differences in serum FFA levels between three above-mentioned groups.Results: Significant increase in the levels of C16:1, C18:1, C18:2, C18:3, C20:4, and C22:6 in healthy controls relative to TC patients and BTD patients was observed, and the levels of C16:1, C18:2, C20:4, and C22:6 in BTD patients were significantly decreased relative to TC patients. Receiver operating characteristic (ROC) analysis indicated that a combination of C16:1, C18:2, C20:4, and C22:6 has excellent diagnostic performance to differentiate BTD patients from TC patients, with an area under the ROC curve of 0.857, a sensitivity of 76.8%, and a specificity of 83.7%.Conclusions: Change in serum levels of FFAs is closely correlated with thyroid diseases, and a biomarker panel (C16:1, C18:2, C20:4, and C22:6) should be of benefit to differentiate BTD patients from TC patients.

Uncoupling angiogenesis and inflammation in peripheral artery disease with therapeutic peptide-loaded microgels

Peripheral artery disease (PAD) is characterized by vessel occlusion and ischemia in the limbs. Treatment for PAD with surgical interventions has been showing limited success. Moreover, recent clinical trials with treatment of angiogenic growth factors proved ineffective as increased angiogenesis triggered severe inflammation in a proportionally coupled fashion. Hence, the overarching goal of this research was to address this issue by developing a biomaterial system that enables controlled, dual delivery of pro-angiogenic C16 and anti-inflammatory Ac-SDKP peptides in a minimally-invasive way. To achieve the goal, a peptide-loaded injectable microgel system was developed and tested in a mouse model of PAD. When delivered through multiple, low volume injections, the combination of C16 and Ac-SDKP peptides promoted angiogenesis, muscle regeneration, and perfusion recovery, while minimizing detrimental inflammation. Additionally, this peptide combination regulated inflammatory TNF-α pathways independently of MMP-9 mediated pathways of angiogenesis in vitro, suggesting a potential mechanism by which angiogenic and inflammatory responses can be uncoupled in the context of PAD. This study demonstrates a translatable potential of the dual peptide-loaded injectable microgel system for PAD treatment.

Serum unsaturated free Fatty acids: potential biomarkers for early detection and disease progression monitoring of non-small cell lung cancer.

Background: Lung cancer (LC) is the deadliest cancer, with earlier stage patients having a better opportunity of long-term survival. The goal of this study is to screen less-invasive and efficient biomarkers for early detection of non-small cell LC (NSCLC).Material and Methods: We performed the simultaneous quantitative detection of six serum unsaturated free fatty acids (FFAs, C16:1, C18:3, C18:2, C18:1, C20:4, and C22:6) from 317 healthy controls, 78 patients with benign lung diseases (BLD), and 202 patients with NSCLC using chip-based direct-infusion nanoelectrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (CBDInanoESI-FTICR MS) in the negative ion mode. Multiple point internal standard calibration curves between the concentration ratios of individual fatty acids to internal standards (ISs, C17:1 as IS of C16:1, C18:3, C18:2, and C18:1 and C21:0 as IS of C20:4 and C22:6) and their corresponding intensity ratios were constructed, with correlation coefficient of > 0.99. Mann-Whitney U test was employed to compare the differences in the levels of the FFAs between the patients and healthy controls.Results: Significantly decreased levels of the FFAs in NSCLC patients were observed compared with healthy controls and BLD patients. Receiver operating characteristic curve analysis indicated that a combination of C16:1, C18:1, C18:3, C18:2, C20:4, and C22:6 could excellently differentiate patients with early-stage NSCLC from healthy controls plus BLD patients, with an AUC value of 0.933, a sensitivity of 84.2%, and a specificity of 89.1%. In addition, a biomarker panel (C16:1 and C18:1) was also confirmed preliminarily to monitor disease progression in NSCLC patients treated with icotinib, with a lead time between 8 and 48 weeks relative to clinical medical imaging.Conclusion: A combination of C16:1, C18:1, C18:3, C18:2, C20:4, and C22:6 may be a powerful biomarker panel for the early detection of NSCLC and a combination of C16:1 and C18:1for disease progression monitoring of NSCLC.

南海南部悬浮颗粒物脂肪酸组成分析

Some fatty acids( FA) are specific to individual classes of organisms and attempts to trace organic materials in the ocean. Fatty acid markers have been widely used to trace or confirm predator-prey relationships so as to illustrate the key trophic linkages in the marine ecosystem. Suspended particulate matter from 0m,75 m and 150 m were collected to better understand the fatty acid composition and its fatty acid biomarkers in the southern South China Sea. The cruise was conducted by"NanFeng"RV from February 22 to March 30,initiated by South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences. The range of FA contents were 9.9—15.65 μg /L,10.45 —14.45 μg /L and 9.65—16.45 μg /L at 0 m,75 m and 150 m,respectively. FA contents indicate that vertical change of FA contents was unobvious.Deep chlorophyll a( Chl a) maximum phenomenon was popular in South China Sea,thus FA /Chl a concentration varied in the water column obviously. The ratio of FA /Chl a was higher than 70 at surface and 150m layer,while lower than 30 at75m layer( except A7). Saturated fatty acids( SFA) and mono-unsaturated fatty acids( MUFA) dominated total fatty acid contents in all SPM from each layer. Major SFA was C16: 0,C18: 0 and C12: 0. Major MUFA were C14: 1n3,C16: 1n7 and C16: 1n9,and major polyunsaturated fatty acids( PUFA) were C18: 2n6 and C22: 2n6. Principle components analysis( PCA) revealed that PC1 was positively related to C16:0,C18:0 and C20:0 concentrations,so PC1 maybe the indicator of phytoplankton and debris. PC2 may be represented the information of zooplankton. Within five FA biomarkers,C16: 1n7 /C16: 0 was positively related to C16: 1 /∑FA,while negatively related to ∑C18 /∑FA,so the combination of C16: 1n7 /C16: 0 and C16: 1 /∑FA can be considered as FA biomarker of diatoms,and ∑C18 /∑FA also can be applied to indicate dinoflagellate.

Long-term effects of fatty acids on cell viability and gene expression of neonatal cardiac myocytes

Fatty acids are the most important source of energy for the adult heart. However, cardiac substrate preference changes during development and alters in pathophysiological states. Fatty acids have also been shown to be involved in signal transduction pathways, thereby affecting gene expression in various cell systems. In the present paper the significance of changes in substrate preference and the potential role of fatty acids in signal transduction in the cardiomyocyte are briefly reviewed. Furthermore, the development of a cellular model system, useful in exploring the long-term effects of fatty acids on the normal and hypertrophic cardiomyocyte, is described. Some aspects of this model system are illustrated by showing the effects of different fatty acid species on cell viability and the effects of fatty acids on the expression of heart type fatty acid-binding protein (H-FABP), a 15 kDa protein thought to be involved in intracellular trafficking of fatty acids. To this end primary cultures of rat neonatal ventricular myocytes were kept in defined medium containing various (combinations of) substrates for up to 48 h. First, the effects of prolonged exposure to different fatty acid species, complexed to BSA, on cell viability were investigated. Exposure of the cells to saturated fatty acids (C16:0 or C18:0), but not mono-unsaturated (C16:1 or C18:1) fatty acids, resulted in cell death, as evidenced by the release of intracellular proteins like lactate dehydrogenase. The detrimental effects of saturated fatty acids were nullified by the co-addition of mono-unsaturated fatty acids. Accordingly, the combination of C16:0 C18:1 was used to examine the effects of fatty acids on the expression of H-FABP. Therefore, the cells were incubated with either (i) glucose only, (ii) fatty acids only, or (iii) glucose plus fatty acids. Incubation with fatty acids (with or without glucose) resulted in a nearly four-fold increase of the H-FABP mRNA level. Similarly, at the protein level the cellular H- FABP LDH ratio increased almost two-fold. In hypertrophic cardiomyocytes (stimulated with the α 1-adrenergic agonist phenylephrine) the stimulatory effect of fatty acids on H-FABP expression was mitigated. These findings strongly suggest that fatty acids are able to modulate gene expression in the context of the cardiac muscle cell.

Gas-liquid chromatographic analysis of lenticular triglycerides: I. Total triglycerides

Triglycerides from bovine, rabbit and chicken lenses were characterized according to molecular weight by gas-liquid chromatography. The total triglycerides were separated from the other lipid classes by preparative thin layer chromatography with subsequent elution to obtain purified triglycerides. The molecular weight distribution of the triglycerides and their fatty acid composition were then determined by gas-liquid chromatography. Triglycerides of similar molecular weight were observed in all the species investigated, with C48, C50, C52 and C54 triglycerides comprising greater than 85% of the total. These triglycerides consisted primarily of various combinations of C16 : 0, C16 : 1, C18 : 0, C18 : 1 and C18 : 2 fatty acids. Several unidentified peaks were eluted shortly after the solvent front was noted.