Addition Of Palmitic Acid Research Articles

Identification, tissue expression, and response to various fatty acid inductions of the fabp gene family in grass carp (Ctenopharyngodon idella)

The grass carp (Ctenopharyngodon idella) holds significant economic value worldwide. However, intensive farming practices often result in lipid metabolic disorder of fish. It is essential to explore the influence of pivotal genes in the process of fat accumulation, particularly from the perspective of fat transport. Fatty acid binding proteins (FABPs) are crucial for selectively binding and transporting free fatty acids (FAs) within cells. Nevertheless, the types of fabp genes in the grass carp genome and their responses to various FAs remain unclear. In the current study, exhaustive bioinformatic analysis identified nine fabp genes in the grass carp genome: fabp1a, fabp1b.1, fabp2, fabp3, fabp6, fabp7a, fabp10a, fabp11a, and fabp11b. Although these genes showed widespread expression across various tissues, each gene displayed distinct preferences for specific primary expression tissues. Notably, fabp1a exhibited prominent expression in muscle, fabp1b.1 in abdominal fat, fabp2 and fabp6 in the intestine, fabp7a in the brain, fabp10a in the hepatopancreas, and fabp11a showed maximal expression in the eye. Findings from primary grass carp hepatocytes revealed that the addition of palmitic acid (PA), high concentrations oleic acid (OA) and docosahexaenoic acid (DHA) (400 µM), significantly increased intracellular lipid accumulation. Intriguingly, the addition of PA resulted in increased expression of fabp2 and fabp11a, while DHA significantly enhanced the expression of fabp7a (P < 0.01). Given that the accumulated FAs are primarily saturated fatty acids, these results suggest that regulating fabps, especially fabp2 and fabp11a, could be a strategy to control fat deposition in grass carp.

Potential role of mitochondrial dysfunction in arrhythmogenesis in coronary artery disease

Introduction. Coronary artery disease (CAD) continues to be the most common pathology in the structure of cardiovascular diseases over the past decades, both in Russia and around the world. In the normal condition, the mitochondria of all body cells have the same function capabilities due to the carriage of the same genome. Therefore, it is possible to assess the respiration activity of cardiomyocyte mitochondria by the respiration of mitochondria from peripheral blood leukocytes.Aim: To compare respiratory activity of mitochondria of peripheral blood leukocytes in patients diagnosed with coronary artery disease and coronary artery disease with developed cardiac rhythm disorders (CRD).Material and methods. The studied groups included 45 patients with CAD without CRD and 39 patients with CAD complicated by CRD. Mitochondria were isolated from peripheral blood leukocytes by differential centrifugation. The rate of oxygen loss in pyruvate-malate and succinate incubation buffers was measured when isolated mitochondria were introduced, as well as when palmitic acid was added to the medium. Oxygen consumption rate for the V3 (active phosphorylating) and V4 (nonphosphorylating) metabolic sates was determined, and on their basis respiratory control coefficient was calculating using the formula V3/V4. Statistical data processing was carried out using STATISTICA 13.0 software.Results. Oxygen consumption rate in mitochondria of patients with uncomplicated CAD and CAD with CRD had no significant differences in either pyruvate-malate or succinate buffers. When palmitic acid was added to the incubation medium, the mitochondria of CAD patients without CRD significantly increased oxygen consumption rate in both incubation media. Mitochondria of CAD patients with CRD did not change oxygen consumption rate in both metabolic states after the addition of palmitic acid in incubation media.Conclusion. On the basis of the data obtained, it can be concluded that the function capabilities of mitochondria in the complicated course of CAD has been exhausted, which manifests itself in the inability to increase ATP synthesis in response to the introduction of additional substrates.

Nutrient-Dependent Mitochondrial Fission Enhances Osteoblast Function

Background: The bone synthesizing function of osteoblasts (OBs) is a highly demanding energy process that requires nutrients. However, how nutrient availability affects OBs behavior and bone mineralization remain to be fully understood. Methods: MC3T3-E1 cell line and primary OBs (OBs) cultures were treated with physiological levels of glucose (G; 5.5 mM) alone or with the addition of palmitic acid (G+PA) at different concentrations. Mitochondria morphology and activity were evaluated by fluorescence microscopy, qPCR, and oxygen consumption rate (OCR) measurement, and OBs function was assessed by mineralization assay. Results: The addition of non-lipotoxic levels of 25 μM PA to G increased mineralization in OBs. G+25 μM PA exposure reduced mitochondria size in OBs, which was associated with increased activation of dynamin-related protein 1, a mitochondrial fission protein, enhanced mitochondria OCR and ATP production, and increased expression of oxidative phosphorylation genes. Treatment with Mdivi-1, a putative inhibitor of mitochondrial fission, reduced osteogenesis and mitochondrial respiration in OBs. Conclusions: Our results revealed that OBs function was enhanced in the presence of glucose and PA at 25 μM. This was associated with increased OBs mitochondrial respiration and dynamics. These results suggest a role for nutrient availability in bone physiology and pathophysiology.

Palmitic acid coating of allogeneic cancellous bone for local antibiotic treatment: A porcine impaction bone grafting model

IntroductionThe worldwide rising number of joint replacements results in increasing revision surgery including a relevant portion of septic loosening accompanied by bone deficiencies. Loading of allogeneic bone with antibiotics provides high local antibiotic concentrations and might eradicate bacteria which appear resistant to systemic antibiotic application. Hydrophobic palmitic acid was shown to be a suitable carrier for antibiotics and prevents biofilm. MethodsCancellous bone derived from 6 to 7 months old piglets was used for a standardized in vitro impaction bone grafting model according to previous studies. The specimens were either thermodisinfected or remained native and palmitic acid with one third and two third partial weight were added and compared with control. Shear force at the interface prosthesis to cement and between cement and bone was measured. The relative micromovements were measured with 6 inductive sensors with a resolution of 0.1 μm at three different measuring heights up to a maximum movement of 150 μm between cement and bone. Taking into account the corresponding applied torque the measured values were normalized in μm/Nm. Statistical analysis was done with SPSS Statistics® Version 26.0 IBM. ResultsSmallest movement was measured for thermodisinfected cancellous bone and a not significant decrease of shear force resistance with addition of palmitic acid was found since supplementing native cancellous bone reduced shear force resistance significantly depending on the weight percentage of palmitic acid. ConclusionSupplementation of porcine cancellous bone with palmitic acid did not significantly reduce shear force resistance of thermodisinfected bone since adding palmitic acid to native bone decreased it significantly depending on the volume added. Palmitic acid seems to be a suitable coating for allogeneic cancellous bone to deliver high local antibiotic concentrations and thermodisinfected cancellous bone might be able to store larger volumes of palmitic acid than native bone without relevant influence on shear force resistance.

Morin, the PPARγ agonist, inhibits Th17 differentiation by limiting fatty acid synthesis in collagen-induced arthritis.

T helper (Th) 17 cells highly contribute to the immunopathology of rheumatoid arthritis. Morin, a natural flavonoid, owns well anti-arthritic action but unclear effect on Th17 differentiation. This study tried to solve this issue and explore the mechanisms in view of cellular metabolism. Naïve CD4+ T cells were treated with anti-CD3/CD28 along with Th17-inducing cytokines. Morin was shown to block Th17 differentiation without affecting cell viability even when Foxp3 was dampened. The mechanisms were ascribed to the limited fatty acid synthesis by restricting FASN transcription, as indicated by metabolomics analysis, nile red staining, detection of triglycerides, FASN overexpression, and addition of palmitic acid. Moreover, morin had slight effect on cell apoptosis and protein palmitoylation during Th17 differentiation, but blocked the binding of RORγt to promoter and CNS2 region of Il17a gene. Oleic acid rescued the inhibition of morin on RORγt function, and Th17-inducing cytokines could not induce RORγt function in SCD1-defficient cells, suggesting that oleic acid but not palmitic acid was the direct effector in the action of morin. Then, PPARγ was identified as the target of morin, and GW9662 or PPARγ CRISPR/Cas9 KO plasmid weakened its above-mentioned effects. The transrepression of FASN by morin was owing to physical interaction between PPARγ and Sp1, and the importance of Sp1 in Th17 differentiation was confirmed by siSp1. Finally, the effects and mechanisms for morin-dampened Th17 responses were confirmed in collagen-induced arthritis (CIA) mice. Collectively, morin inhibited Th17 differentiation and alleviated CIA by limiting fatty acid synthesis subsequent to PPARγ activation.

Reversal of stem cell‐derived hypertrophic adipocytes mediated by photobiomodulation (1064 nm)

AbstractHypertrophic adipocytes have been implicated in the progression of obesity‐induced complications. These enlarged fat cells cause release of free fatty acids and inflammatory cytokines, promote hypoxia and fibrosis and limit insulin sensitivity. Photobiomodulation (PBM), or the influence of light on biological tissues, has previously been demonstrated to reduce lipid accumulation in stem cells undergoing adipogenesis. Here, we characterize the effect of PBM on an in vitro hypertrophic obesity model and its influence on reduction of lipids and restoration to normal adipocyte function. Adipose‐derived stem cells were induced to hypertrophy with the addition of palmitic acid (PA) and was confirmed with fluorescent imaging of lipid content, and functional changes such as glucose uptake. Glucose transport into the cell was diminished and the expression of glucose transporter GLUT4 was downregulated. PBM‐treated hypertrophic cells (1064 nm and 17.6 J/cm2 every day for 7 days following addition of PA) decreased the lipid levels in hypertrophic adipocytes, restored the GLUT4 protein expression and enhanced glucose transport. Taken together, PBM is shown capable of restoring the cellular morphology and function of hypertrophic cells. This could have important clinical implication for the development of laser‐based potential therapeutic treatment of complications due to metabolic syndrome.

Physicochemical characteristic of modified edible film made from gelatine of sea bass (Lates calcarifer) residue with palmitic acid and soybean protein isolate treatment

The use of fish waste as raw material in producing gelatin becomes important when it is related to the halal perspective. Sea bass skin contains high protein, especially collagen protein which can be used as the main component of edible film. An edible film made from gelatin of fish skin usually lacks water vapour transmission due to its hydrophilic properties. Modification of formula in producing an edible film from fish gelatin with the addition of lipid such palmitic acid (PA treatment) and soybean protein isolate (SPI treatment) can affect the characteristics of the edible film resulted. This study was aimed to evaluate the chemical (solubility, water vapour permeability) and physical characteristics (thickness, tensile strength, per cent elongation, SEM) of an edible film made from gelatin with the addition of palmitic acid and soybean protein isolate with different concentration. The results showed that edible films with SPI addition yield the best film thickness (&lt;2.5 mm) while PA addition resulted in better tensile strength of the films (&gt;3.92 MPa) according to the Japanese Industrial Standard. The per cent elongation of both films with PA (21.3-34.44%) and SPI (36.06-117.53%) addition decreased with increasing concentration, but SPI generally gave higher elongation compared to PA. The addition of SPI gave lower solubility and higher water vapour transmission values compared to PA, however, the addition of 4.5% of PA gave the lowest water vapour transmission (5.416 g/m2 h). Therefore, the best treatment in this study was concluded to be 4% of SPI addition based on the best thickness, per cent elongation, water vapour transmission and solubility results.

Long chain fatty acid affects excited state branching in bilirubin-human serum protein Complex

After binding to human serum albumin, bilirubin could undergo photoisomerization and photo-induced cyclization process. The latter process would result the formation of a product, named as lumirubin. These photo induced behaviors are the fundamental of clinical therapy for neonatal jaundice. Previous studies have reported that the addition of long chain fatty acids is beneficial to the generation of lumirubin, yet no kinetic study has revealed the mechanism behind. In this study, how palmitic acid affects the photochemical reaction process of bilirubin in Human serum albumin (HSA) is studied by using femtosecond transient absorption and fluorescence up-conversion techniques. With the addition of palmitic acid, the excited population of bilirubin prefers to return to its hot ground state (S0) through a 4 ps decay channel rather than the intrinsic ultrafast decay pathways (&amp;lt;1 ps). This effect prompts the Z-Z to E-Z isomerization at the S0 state and then further increases the production yield of lumirubin. This is the first time to characterize the promoting effect of long chain fatty acid in the process of phototherapy with femtosecond time resolution spectroscopy and the results can provide useful information to benefit the relevant clinical study.

Effect of Addition Palmitic Acid and Vitamin E to the Tris-Egg Yolk Diluter in Canine Semen Cryopreservation

Background: Canine sperm is a very delicate cell that is quite susceptible to oxidative stress since the cytoplasm is restricted and features little antioxidant reserves. Furthermore, the sperm membrane has some polyunsaturated fatty acids sensitive to lipid peroxidation, which makes it important to addition antioxidant substances to the diluter aiming at decreasing such stress to the sperm cell, particularly during seminal cryopreservation. Several antioxidants have been used in this process in some domestic animal’s species, however, the use of palmitic acid has been little reported in works on cryopreservation of semen of the canine species. Hence, this study aimed to assess the effect of addition antioxidants palmitic acid and vitamin E to the Tris-egg yolk diluter on the semen quality of dogs after thawing.Materials, Methods &amp; Results: Samples were collected from the ejaculates of 4 adult dogs, apparently healthy, of the American Pit Bull Terrier breed of kennels in the city of Teresina, PI, places where the pre-freezing procedures of the dog’s semen were performed. The samples were diluted in Tris citric acid fructose (3.28 g Tris-hydroxymethyl-aminomethane, 1.78 g citric acid monohydrate and 1.25 g D-fructose), dissolved in 100 mL distilled water, and added 20% egg yolk and 6% glycerol, at the concentration of 100x106 sptz/mL. The semen samples were divided into 3 mL aliquots to form 3 experimental groups: G1 - Only Tris-egg yolk (Control group); G2 - Tris-egg yolk + 100 µM palmitic acid; and G3 - Tris-egg yolk + 116 µM vitamin E. Semen was collected weekly over a period of little over 2 months. After thawing, thermorresistance test (TTR) was carried out at 0, 30, 60, and 90 min to assess spermatics motility and vigor, in addition to analysis of integrity of plasma membrane, acrosomal membrane and mitochondrial activity of the sperm, using fluorescent probes. These assessments were performed out at the Animal Reproduction Biotechnology Laboratory (LBRA/UFPI). In the TTR, G2 and G3 didn´t exhibit significant results for spermatics motility or vigor when compared with the control group. The palmitic acid and vitamin E also had no significant effects on the parameters of acrosomal membrane integrity or mitochondrial activity. However, sperm cryopreserved with the addition of palmitic acid exhibited significant differences for plasma membrane integrity, providing greater protection to the sperm cells in G2.Discussion: The palmitic acid is one of the most saturated fatty acids in human semen, with reports of great proportions also in the seminal plasma of dogs. Its main role is to protect the plasma membrane from external damage, improving viability and fertility of the sperm after cryopreservation. Data is scarce in the literature on the composition of fatty acids in canine semen and regarding the use of palmitic acid as a seminal antioxidant in that species, which grants further studies aiming to investigate such valuable information for canine reproduction. It is concluded that addition palmitic acid at 100 µM concentration to the Tris-egg yolk diluter was able to preserve the integrity of the plasma membrane during the process of cryopreservation of canine semen.Keywords: dog, semen, antioxidants, cryopreservation.Descritores: cão, sêmen, antioxidantes, criopreservação.

Skirmishing MDR strain of Candida albicans by effective antifungal CeO2 nanostructures using Aspergillus terreus and Talaromyces purpurogenus

Emerging antibiotics resistance fungal infectionsis a major global health problem and new antifungal formulations are direly needed to fight drug resistant Candida albicans strains. This study is aimed to synthesize effective antifungal nanostructures of cerium oxide (CeO2) using culture filtrates of two common fungal strains Aspergillus terreus and Talaromyces pupureogenus. The fungal strains used in the synthesis were identified by 18S rRNA gene sequencing and deposited to NCBI GenBank with the accession number of MN099077 and MN121629, respectively. The biofabricated CeO2 NPs were characterized by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). Pure CeO2 nanoparticles (NPs) synthesized using Aspergillus terreus culture filtrate were depicted spherical morphology with average size of 28.5 nm. The CeO2NPs synthesized using Talaromyces pupureogenus revealed the presence of nanosponges with average size of 21.4 nm. Gas chromatography mass spectrometry of culture filtrates of respective strains indicated the presence of ethanol, 1-propanol and tri-chloromethane in culture filtrate of Aspergillus terreus and with addition of palmitic acid in Talaromyces pupureogenus culture filtrate which may have a function as bio reducers and capping agents. Dose dependent anticandidal activity of CeO2 NPs using various different concentrations (100, 200, 300, 600 μg ml−1) synthesized by both fungal strains was observed by disc diffusion assay against Candida albicansas evidenced by increase in size of zone of inhibitions with increasing concentration of CeO2NPs. Further in-vitro and in-vivo experiments are required to access the potential of CeO2 NPs for controlling Candida albicans strains.

Subcellular localisation of FLAG tagged enzymes of the dynamic protein S-palmitoylation cycle of Trypanosoma cruzi epimastigotes.

Dynamic S-palmitoylation of proteins is the addition of palmitic acid by zDHHC palmitoyl transferases (PATs) and depalmitoylation by palmitoyl protein thioesterases (PPTs). A putative PAT (TcPAT1) has been previously identified in Trypanosoma cruzi, the etiological agent of Chagas disease. Here we analyse other 14 putative TcPATs and 2 PPTs in the parasite genome. T. cruzi cell lines expressing TcPATs and TcPPTs plus a FLAG tag at the C terminus were produced for most enzymes, with positive detection by indirect immunofluorescence. Overexpressed TcPATs were mostly found as single spots at the parasite anterior end, while the TcPPTs were dispersed throughout the parasite body.

MiR‐21‐5p Regulation of Cardiac Fatty Acid Oxidation and Mitochondrial Respiration

Cardiovascular related pathologies are the leading single cause of death in chronic kidney disease (CKD) patients. Previously, we found that 5/6 nephrectomy model of CKD leads to an upregulation of miR‐21‐5p in the left ventricle, targeting peroxisome proliferator‐activated receptor alpha (PPARα) and altering the expression of numerous transcripts involved with fatty acid oxidation and glycolysis. We also observed that overexpression of miR‐21‐5p attenuates both lipid content and lipid peroxidation in H9C2 (cardiomyoblast, ATCC) cells. These findings lead us to hypothesize that miR‐21‐5p regulates cardiac mitochondrial respiration and fatty acid oxidation. In this study we utilized the Seahorse XF Fatty Acid Oxidation Assay (Agilent) to evaluate the potential for knockdown or overexpression of miR‐21‐5p to impact oxygen consumption rate in the H9C2 cells. Cells were plated into 96 well Seahorse assay plates and transfected with anti‐miR‐21‐5p (40nM), pre‐miR‐21‐5p (20nM) or appropriate scrambled oligonucleotide controls. After a 24‐hour incubation medium was changed from 4.5g/L glucose to 1.0g/L glucose and incubated for an additional 24 hours. In preparation for the Seahorse Assay, medium was changed and half of the cells from each transfection group were treated with etomoxir, a compound that prevents fatty acid oxidation, for 30 minutes. This was followed by addition of palmitic acid (PA), a saturated fatty acid, to half of the cells of each transfection/treatment group. When we compare mitochondrial respiration rate (oxygen consumption rate normalized by total cellular protein in that sample) we find that both anti‐miR‐21‐5p and pre‐miR‐21‐5p increase basal and maximal respiration rates. However, it should be noted that maximal respiration is significantly higher with anti‐miR‐21‐5p than with pre‐21‐5p (172% vs. 139% of control, respectively). Similarly, we find that non‐mitochondrial respiration, basal respiration, maximal respiration, ATP production and proton leak are all significantly increased in response to either pre‐miR‐21‐5p or anti‐miR‐21‐5p when compared to their respective control treatment. Neither etomoxir, nor PA had any impact on oxygen consumption rate in pre‐miR‐21‐5p treated cells, suggesting that fatty acid was not contributing significantly to the oxygen consumption when miR‐21‐5p is elevated. Conversely, anti‐miR‐21‐5p cell transfected cells exhibited reduced oxygen consumption rate with both etomoxir and PA, with PA treated cells exhibiting signs of lipotoxicity. To determine if miR‐21‐5p manipulation altered mitochondria oxidative capacity, we transfected and treated cells as described above, and then measured the amount of lactate produced over a one‐hour using the Lactate Glo Assay (Promega). Cellular lactate production was not increased, and tended to decrease in both pre‐miR‐21‐5p and anti‐miR‐21‐5p treatment groups, when compared to controls, suggesting oxidative capacity is not impaired in either condition. In sum these results suggest that overexpression and suppression of miR‐21‐5p augment mitochondrial respiration in H9C2 cells through different mechanisms. Together with previous findings, our results indicate that the overexpression of miR‐21‐5p reduces cellular fatty acid utilization, potentially shifting cellular metabolism toward reliance on the glycolytic pathway.Support or Funding InformationR01HL128332This 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 annealing on the functionality of Bambara groundnut (Vigna subterranea) starch–palmitic acid complex

SummaryBambara groundnut is an underutilised African leguminous crop. This study investigated the effect of annealing on the complexing ability and functionality of Bambara groundnut starch with palmitic acid. Corn starch was included as the reference. Annealing created cracks and pores on the surface of Bambara groundnut and corn starches, respectively. Bambara groundnut starch had significantly higher amylose content, higher peak and final viscosities than corn starch. The peak viscosities of native Bambara groundnut and corn starches significantly reduced with palmitic acid addition. Greater reduction in peak viscosities was observed when the annealed starches were complexed with palmitic acid, suggesting that more palmitic acid was complexed after annealing. This was confirmed by XRD peaks and melting enthalpies. Pasting of native Bambara groundnut and corn starches with palmitic acid resulted in the formation of type I V‐amylose complexes, while type II complexes were formed from annealed starches pasted with palmitic acid.

The Myxobacterial Metabolite Soraphen A Inhibits HIV-1 by Reducing Virus Production and Altering Virion Composition.

Soraphen A is a myxobacterial metabolite that blocks the acetyl-coenzyme A carboxylase of the host and was previously identified as a novel HIV inhibitor. Here, we report that soraphen A acts by reducing virus production and altering the gp120 virion content, impacting entry capacity and infectivity. These effects are partially reversed by addition of palmitic acid, suggesting that inhibition of HIV envelope palmitoylation is one of the mechanisms of antiviral action.

On the “swelling” of mitochondria under palmitic acid, calcium, and hypotension treatment

The high-amplitude swelling of mitochondria is critically considered. In contrast to numerous statements by some authors about a marked swelling of isolated liver mitochondria under the influence of palmitic acid, calcium ions, or hypotension, we have shown that mitochondria are generally not subject to highamplitude swelling. According to optical-microscopy data even during long-lasting incubation (in distilled water) where full hypotension takes place, the size of liver mitochondria (approximately 1 µm) can be enlarged by no more than by 40%. Under short-lasting hypotension or the addition of palmitic acid the mitochondrial diameter becomes greater by only 20% or remains virtually unchanged. The light scattering of the mitochondrial suspension measured using a photometer according to the decrease in optical density declines by 2.5 times. A decrease in the light scattering in hypotension or via the addition of palmitic acid or calcium (in an isotonic medium) occurs because of damage (even destruction) to the outer membrane, rather than due to the swelling of mitochondria, as was previously believed. The inner membrane is not significantly expanded. The destruction of the outer membrane reduces the probability of light scattering by each mitochondrion at the boundary layer of the water/membrane interface. Release of substances from the matrix resulting in a decrease of its refractive index may additionally contribute to the decrease in light scattering. Palmitic acid and calcium (at concentrations of 10 to 100 µM) cause permeabilization and disruption of the outer membrane gradually, over several minutes. Full hypotension activates this process very rapidly, viz., within a fraction of a second. Under low ionic-strength conditions, the addition of calcium leads to neutralization of negative charges on the membrane surface, which induces aggregation of mitochondria, thus enhancing light scattering and creating the illusion of mitochondrial swelling.

Simultaneous quantification of salivary 3-hydroxybutyrate, 3-hydroxyisobutyrate, 3-hydroxy-3-methylbutyrate, and 2-hydroxybutyrate as possible markers of amino acid and fatty acid catabolic pathways by LC-ESI-MS/MS.

We have developed a highly sensitive and specific method for quantification of salivary 3-hydroxybutyrate (3HB), 3-hydroxyisobutyrate (3HIB), 3-hydroxy-3-methylbutyrate (3HMB) and 2-hydroxybutyrate (2HB), which could be new non-invasive biomarkers for catabolic pathways of fatty acids/ketogenic amino acids, valine, leucine, and methionine/threonine/α-ketobutyrate, respectively. The four hydroxybutyrates (3HB, 3HIB, 3HMB, and 2HB) were extracted from 5 µl of saliva, converted to 2-pyridylmethyl (2PM) ester derivatives, and measured by liquid chromatography–tandem mass spectrometry in positive electrospray ionization mode. [13C4]3HB was used as an internal standard. The detection limits for the 2PM esters were <1 pg (7.9–9.6 fmol) on-column (signal-to-noise ratio = 3). Reproducibilities and recoveries of the hydroxybutyrates were validated according to one-way layout and polynomial equation, respectively. The variances between sample preparations and between measurements were calculated to be 0.45–5.28 and 0.54–3.45 %, respectively. Experiments performed using 5 µl of saliva spiked with 3.8–154.4 pmol of the four hydroxybutyrates gave recoveries of 98.5 to 108.8 %, with a mean recovery of 104.1 %. In vitro experiments in hepatocytes or skeletal muscle cells showed that addition of palmitic acid, valine, leucine or α-ketobutyrate to culture medium markedly increased the targeted hydroxybutyrate concentrations. The salivary concentration of each targeted hydroxybutyrate was positively correlated with that in serum, and the salivary levels were elevated in patients with liver cirrhosis, which is characterized by upregulated catabolism of lipids and amino acids. The proposed method is useful for quantification of salivary 3HB, 3HIB, 3HMB, and 2HB for monitoring of catabolic activities of amino acids and fatty acids.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-015-1304-0) contains supplementary material, which is available to authorized users.

Metal–Organic Frameworks Encapsulated in Photocleavable Capsules for UV-Light Triggered Catalysis

A smart catalyst that can be activated by UV-light was prepared by encapsulating metal–organic framework (MOF) particles in microcapsules with a photocleavable polyurea shell. Addition of palmitic acid as a modulator during the synthesis of the Fe-terephthalate MOF MIL-88B(Fe) yields MOF particles with a hydrophobic surface. These particles can be successfully encapsulated via interfacial polymerization, as proven by a combination of powder X-ray diffraction (PXRD), scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) techniques. By the incorporation of photolabile groups in the polymer shell, UV light can be employed to trigger degradation of the capsules, releasing the catalytic load. Catalyst activation triggered by UV light was monitored by recording the rates for catalytic tetramethylbenzidine (TMB) oxidation with H2O2. The reactivity could be controlled by tuning irradiation time, UV intensity, or MOF loading. The activity of the capsules was up to 90 times higher after irradiation.

MP31-06 DIET INDUCED ALTERATION OF FATTY ACID SYNTHASE IS INVOLVED IN PROSTATE CANCER PROGRESSION

You have accessJournal of UrologyProstate Cancer: Basic Research II1 Apr 2014MP31-06 DIET INDUCED ALTERATION OF FATTY ACID SYNTHASE IS INVOLVED IN PROSTATE CANCER PROGRESSION Mingguo Huang, Shintaro Narita, Norihiko Tsuchiya, Takamitsu Inoue, Shigeru Satoh, Hiroshi Nanjo, Takehiko Sasaki, and Tomonori Habuchi Mingguo HuangMingguo Huang More articles by this author , Shintaro NaritaShintaro Narita More articles by this author , Norihiko TsuchiyaNorihiko Tsuchiya More articles by this author , Takamitsu InoueTakamitsu Inoue More articles by this author , Shigeru SatohShigeru Satoh More articles by this author , Hiroshi NanjoHiroshi Nanjo More articles by this author , Takehiko SasakiTakehiko Sasaki More articles by this author , and Tomonori HabuchiTomonori Habuchi More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.915AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES High-fat diet (HFD) or obesity is strongly suggested to influence prostate cancer (PCa) progression, but the underlying mechanism is not well defined. Fatty acid synthase (FASN) is a cytosolic metabolic enzyme catalyzing de novo biosynthesis of fatty acid. METHODS In the present study, we investigated the role of FASN on PCa progression in the LNCaP xenograft mouse fed HFD or low-fat diet (LFD). RESULTS The tumor growth was increased in the HFD group than LFD group (p = 0.025). The mRNA expression of FASN and SREBP-1, which encodes Sterol regulatory element-binding protein-1 (SREBP-1) were increased in the xenograft tumor by 1.8- and 2.1-fold, respectively (p < 0.05). The serum level of FASN was significantly lower in the HFD group than in the LFD group (p = 0.026) and correlated inversely with the tumor volumes (total, r = 0.642, p = 0.022; HFD group, r = −0.618, p = 0.024; LFD group, r = −0.439, p = 0.154; respectively). Upregulation of P-AKT and P-ERK, and downregulation of P-AMPK were observed in xenograft tumors of the HFD group than LFD group. The extracellular release of FASN protein in vitro and AMPK activation was induced in the PCa cells by inhibition of the PI3K and MAPK signaling. In addition, pharmacological activation of AMPK stimulated the extracellular FASN release along with the downregulation of FASN and SREBP-1 in the PCa cells in dose-dependent manner. Cell proliferation was significantly decreased in the PCa cells by inhibition of FASN, and the decreased effect was rescued by addition of palmitic acid. Clinically, imuunohistological evaluation of surgical specimens showed that the expression of FASN was markedly decreased in the PCa response to hormone and chemotherapy. CONCLUSIONS In conclusion, HFD increased the FASN expression and inhibited the extracellular release of FASN, which may be one of important mechanism in HFD or obesity associated PCa progression. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e324 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Mingguo Huang More articles by this author Shintaro Narita More articles by this author Norihiko Tsuchiya More articles by this author Takamitsu Inoue More articles by this author Shigeru Satoh More articles by this author Hiroshi Nanjo More articles by this author Takehiko Sasaki More articles by this author Tomonori Habuchi More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Dietary oleic acid increases m2 macrophages in the mesenteric adipose tissue.

Several studies have implicated fatty-acids as inflammatory regulators, suggesting that there may be a direct role for common dietary fatty-acids in regulating innate immune cells. In humans, a single high-fat meal increases systemic cytokines and leukocytes. In mice, short term high-fat feeding increases adipose tissue (AT) leukocytes and alters the inflammatory profile of AT macrophages. We have seen that short term high fat feeding to C57BL/6J male mice increases palmitic and oleic acid within AT depots, but oleic acid increase is highest in the mesenteric AT (MAT). In vitro, oleic acid increases M2 macrophage markers (CD206, MGL1, and ARG1) in a murine macrophage cell line, while addition of palmitic acid is able to inhibit that increase. Three day supplementation of a chow diet, with oleic acid, induced an increase in M2 macrophage markers in the MAT, but not in the epididymal AT. We tested whether increases in M2 macrophages occur during short term ad lib feeding of a high fat diet, containing oleic acid. Experiments revealed two distinct populations of macrophages were altered by a three day high milk-fat diet. One population, phenotypically intermediate for F4/80, showed diet-induced increases in CD206, an anti-inflammatory marker characteristic of M2 macrophages intrinsic to the AT. Evidence for a second population, phenotypically F4/80HICD11bHI macrophages, showed increased association with the MAT following short term feeding that is dependent on the adhesion molecule, ICAM-1. Collectively, we have shown that short term feeding of a high-fat diet changes two population of macrophages, and that dietary oleic acid is responsible for increases in M2 macrophage polarization.

Rhizosphere effect of Scirpus triqueter on soil microbial structure during phytoremediation of diesel-contaminated wetland

Though phytoremediation has been widely used to restore various contaminated sites, it is still unclear how soil microbial communities respond microecologically to plants and pollutants during the process. In this paper, batch microcosms imitating in situ phytoremediation of petroleum-contaminated wetland by Scirpus triqueter were set up to monitor the influence of plant rhizosphere effect on soil microbes. Palmitic acid, one of the main root exudates of S. triqueter, was added to strengthen rhizosphere effect. Abundances of certain microbial subgroups were quantified by phospholipid fatty acid profiles. Results showed that diesel removal extents were significantly higher in the rhizosphere (57.6±4.2–65.5±6.9%) than those in bulk soil (27.8±6.5–36.3±3.2%). In addition, abundances of saturated, monounsaturated, and polyunsaturated fatty acids were significantly higher (P<0.05) in planted soil than those in the bulk soil. When it was less than 15,000 mg diesel kg soil−1, increasing diesel concentration led to higher abundances of fungi, Gram-positive and Gram-negative bacteria. The addition of palmitic acid amplified the rhizosphere effect on soil microbial populations and diesel removal. Principal component analysis revealed that plant rhizosphere effect was the dominant factor affecting microbial structure. These results provided new insights into plant-microbe-pollutant coactions responsible for diesel degradation, and they were valuable to facilitate phytoremediation of diesel contamination in wetland habitats.