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Porphyromonas gingivalis has been identified as one of the major periodontal pathogens. Activity-directed fractionation and purification processes were employed to identify bioactive compounds from bitter melon leaf. Ethanolic extract of bitter melon leaf was separated into five subfractions by open column chromatography. Subfraction-5-3 significantly inhibited P. gingivalis-induced interleukin (IL)-8 and IL-6 productions in human monocytic THP-1 cells and then was subjected to separation and purification by using different chromatographic methods. Consequently, 5β,19-epoxycucurbita-6,23(E),25(26)-triene-3β,19(R)-diol (charantadiol A) was identified and isolated from the subfraction-5-3. Charantadiol A effectively reduced P. gingivalis-induced IL-6 and IL-8 productions and triggered receptors expressed on myeloid cells (TREM)-1 mRNA level of THP-1 cells. In a separate study, charantadiol A significantly suppressed P. gingivalis-stimulated IL-6 and tumor necrosis factor-α mRNA levels in gingival tissues of mice, confirming the inhibitory effect against P. gingivalis-induced periodontal inflammation. Thus, charantadiol A is a potential anti-inflammatory agent for modulating P. gingivalis-induced inflammation.

Pinolenic acid (PNA) is a rare n-6 polyunsaturated fatty acid (n-6 PUFA) originally identified in pine seeds. Previous studies demonstrated that PNA and its elongation metabolite, Δ7-eicosatrienoic acid (Δ7-ETrA), exerted an anti-inflammatory effect in cultured cells by suppressing prostaglandin E2 (PGE2) production. The objective of this study was to further examine the in vivo anti-inflammatory properties of PNA. Using human THP-1 macrophage, we first confirmed that incorporation of PNA into cellular phospholipids suppressed the production of interleukin-6 (IL-6) (by 46%), tumor necrosis factor-α (TNF-α) (by 18%), and prostaglandin E2 (PGE2) (by 87%), and the expression of type-2 cyclooxygenase (COX-2) (by 27%). Furthermore, we demonstrated that injection of PNA or Δ7-ETrA suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema, as measured by ear thickness (by 15%) and biopsy weight (by up to 29%). Both PUFA also lowered proportions of infiltrated leukocytes, neutrophils, and macrophages using flow cytometric analysis. Topical application of PNA or Δ7-ETrA on mouse back skin suppressed TPA-induced pro-inflammatory mediator production, including IL-1β, IL-6, TNF-α, and PGE2, as well as the phosphorylation of p38- and JNK-mitogen-activated protein kinase (MAPK), but not that of ERK-MAPK. That no PNA or Δ7-ETrA was detected in the ear disc after the PUFA injection suggests that their anti-inflammatory effect might not be due to fatty acid incorporation, but to modulation of cell signaling. In conclusion, PNA and Δ7-ETrA exerted the in vivo anti-inflammatory effect by suppressing mouse ear edema and dorsal skin inflammation.

Even with increasing evidence for roles of glycolytic enzymes in controlling cancerous characteristics, the best target of candidate metabolic enzymes for lessening malignancy remains under debate. Pyruvate is a main glycolytic metabolite that could be mainly converted into either lactate by Lactate Dehydrogenase A (LDHA) or acetyl-CoA by Pyruvate Dehydrogenase E1 component α subunit (PDHA1) catalytic complex. In tumor cells, accumulating lactate is produced whereas the conversion of pyruvate into mitochondrial acetyl-CoA is less active compared with their normal counterparts. This reciprocal molecular association makes pyruvate metabolism a potential choice of anti-cancer target. Cellular and molecular changes were herein assayed in Head and Neck Squamous Cell Carcinoma (HNSCC) cells in response to LDHA and PDHA1 loss in vitro, in vivo and in clinic. By using various human cancer databases and clinical samples, LDHA and PDHA1 levels exhibit reversed prognostic roles. In vitro analysis demonstrated that decreased cell growth and motility accompanied by an increased sensitivity to chemotherapeutic agents was found in cells with LDHA loss whereas PDHA1-silencing exhibited opposite phenotypes. At the molecular level, it was found that oncogenic Protein kinase B (PKB/Akt) and Extracellular signal-regulated kinase (ERK) singling pathways contribute to pyruvate metabolism mediated HNSCC cell growth. Furthermore, LDHA/PDHA1 changes in HNSCC cells resulted in a broad metabolic reprogramming while intracellular molecules including polyunsaturated fatty acids and nitrogen metabolism related metabolites underlie the malignant changes. Collectively, our findings reveal the significance of pyruvate metabolic fates in modulating HNSCC tumorigenesis and highlight the impact of metabolic plasticity in HNSCC cells.

Cutibacterium acnes (formerly Propionibacterium acnes) is one of the major bacterial species responsible for acne vulgaris. Numerous bioactive compounds from Momordica charantia Linn. var. abbreviata Ser. have been isolated and examined for many years. In this study, we evaluated the suppressive effect of two cucurbitane-type triterpenoids, 5&beta, 19-epoxycucurbita-6,23-dien-3&beta, 19,25-triol (Kuguacin R, KR) and 3&beta, 7&beta, 25-trihydroxycucurbita-5,23-dien-19-al (TCD) on live C. acnes-stimulated in vitro and in vivo inflammatory responses. Using human THP-1 monocytes, KR or TCD suppressed C. acnes-induced production of interleukin (IL)-1&beta, IL-6 and IL-8 at least above 56% or 45%, as well as gene expression of these three pro-inflammatory cytokines. However, a significantly strong inhibitory effect on production and expression of tumor necrosis factor (TNF)-&alpha, was not observed. Both cucurbitanes inhibited C. acnes-induced activation of the myeloid differentiation primary response 88 (MyD88) (up to 62%) and mitogen-activated protein kinases (MAPK) (at least 36%). Furthermore, TCD suppressed the expression of pro-caspase-1 and cleaved caspase-1 (p10). In a separate study, KR or TCD decreased C. acnes-stimulated mouse ear edema by ear thickness (20% or 14%), and reduced IL-1&beta, expressing leukocytes and neutrophils in mouse ears. We demonstrated that KR and TCD are potential anti-inflammatory agents for modulating C. acnes-induced inflammation in vitro and in vivo.

Cutibacterium acnes (formerly Propionibacterium acnes) is one of the major bacterial species responsible for acne vulgaris. Numerous bioactive compounds from Momordica charantia Linn. var. abbreviata Ser. have been isolated and examined for many years. In this study, we evaluated the suppressive effect of two cucurbitane-type triterpenoids, 5β,19-epoxycucurbita-6,23-dien-3β,19,25-triol (Kuguacin R; KR) and 3β,7β,25-trihydroxycucurbita-5,23-dien-19-al (TCD) on live C. acnes-stimulated in vitro and in vivo inflammatory responses. Using human THP-1 monocytes, KR or TCD suppressed C. acnes-induced production of interleukin (IL)-1β, IL-6 and IL-8 at least above 56% or 45%, as well as gene expression of these three pro-inflammatory cytokines. However, a significantly strong inhibitory effect on production and expression of tumor necrosis factor (TNF)-α was not observed. Both cucurbitanes inhibited C. acnes-induced activation of the myeloid differentiation primary response 88 (MyD88) (up to 62%) and mitogen-activated protein kinases (MAPK) (at least 36%). Furthermore, TCD suppressed the expression of pro-caspase-1 and cleaved caspase-1 (p10). In a separate study, KR or TCD decreased C. acnes-stimulated mouse ear edema by ear thickness (20% or 14%), and reduced IL-1β-expressing leukocytes and neutrophils in mouse ears. We demonstrated that KR and TCD are potential anti-inflammatory agents for modulating C. acnes-induced inflammation in vitro and in vivo.

Cutibacterium acnes (formerly Propionibacterium acnes) is a key pathogen involved in the development and progression of acne inflammation. The numerous bioactive properties of wild bitter melon (WBM) leaf extract and their medicinal applications have been recognized for many years. In this study, we examined the suppressive effect of a methanolic extract (ME) of WBM leaf and fractionated components thereof on live C. acnes-induced in vitro and in vivo inflammation. Following methanol extraction of WBM leaves, we confirmed anti-inflammatory properties of ME in C. acnes-treated human THP-1 monocyte and mouse ear edema models. Using a bioassay-monitored isolation approach and a combination of liquid&ndash, liquid extraction and column chromatography, the ME was then separated into n-hexane, ethyl acetate, n-butanol and water-soluble fractions. The hexane fraction exerted the most potent anti-inflammatory effect, suppressing C. acnes-induced interleukin-8 (IL-8) production by 36%. The ethanol-soluble fraction (ESF), which was separated from the n-hexane fraction, significantly inhibited C. acnes-induced activation of mitogen-activated protein kinase (MAPK)-mediated cellular IL-8 production. Similarly, the ESF protected against C. acnes-stimulated mouse ear swelling, as measured by ear thickness (20%) and biopsy weight (23%). Twenty-four compounds in the ESF were identified using gas chromatograph&ndash, mass spectrum (GC/MS) analysis. Using co-cultures of C. acnes and THP-1 cells, &beta, ionone, a compound of the ESF, reduced the production of IL-1&beta, and IL-8 up to 40% and 18%, respectively. &beta, ionone also reduced epidermal microabscess, neutrophilic infiltration and IL-1&beta, expression in mouse ear. We also found evidence of the presence of anti-inflammatory substances in an unfractionated phenolic extract of WBM leaf, and demonstrated that the ESF is a potential anti-inflammatory agent for modulating in vitro and in vivo C. acnes-induced inflammatory responses.

Ling-Zhi-8 (LZ-8), a 13-kDa protein from mycelia of Ganoderma lucidum, exerts various beneficial functions in mammalian cells. Using murine BV-2 microglial cells, the objective of this study was to determine how LZ-8 modulates the production of pro-inflammatory mediators. Incubation of cells with LZ-8 (up to 10 µg/ml) had no negative effect on cell proliferation. LZ-8 significantly reduced the lipopolysaccharide-stimulated production of nitric oxide, prostaglandin E2 and interleukin-6 up to 31%, 39% and 58%, respectively (p

Demand for long chain ω-3 fatty acids from non-fish source for vegetarians has increased recently. Marine microalgae are the primary producers of EPA/DHA and promising alternatives for fish oil. Tropical areas have abundant sunlight throughout the year for microalgal cultivation but this practice can be hindered by high temperature. Discovery of heat-tolerant marine microalgae that can synthesize EPA/DHA will solve these problems. A new species of microalga was isolated from a high temperature lagoon and identified as Tetraselmis sp. DS3. These cells could grow at 40 °C, the highest temperature for marine microalgal growth ever reported. Its ω-3 fatty acids and EPA accounted for 33 and 10% of total lipids, respectively, grown in nitrogen-depleted conditions. These cells also accumulated more than 5% β-carotene and 0.48% lutein in biomass. This new microalga can be cultivated for long chain ω-3 fatty acids and lutein production in the tropical areas.

Acne vulgaris (acne) is a common inflammatory skin disorder, and Propionibacterium acnes plays a major role in the development and progression of acne inflammation. Herbs possessing antimicrobial and anti-inflammatory activity have been applied as a medical option for centuries. In this study, we examined the suppressive effect of ethanolic oregano (Origanum vulgare) extract on live P. acnes-induced in vivo and in vitro inflammation. Following ethanol extraction of oregano leaves, four compounds with strong antioxidant activity, including rosmarinic acid, quercetin, apigenin, and carvacrol, were identified by high-performance liquid chromatography. Using the mouse ear edema model, we demonstrated that ethanol oregano extracts (EOE) significantly suppressed P. acnes-induced skin inflammation, as measured by ear thickness (32%) and biopsy weight (37%). In a separate study, using the co-culture of P. acnes and human THP-1 monocytes, EOE reduced the production of interleukin (IL)-8, IL-1&beta, and tumor necrosis factor (TNF)-&alpha, up to 40%, 37%, and 18%, respectively, as well as the expression of these three pro-inflammatory mediators at the transcriptional level. Furthermore, EOE inhibited the translocation of nuclear factor-kappa B (NF-&kappa, B) into the nucleus possibly by inactivating toll-like receptor-2 (TLR2). The suppressive effect of EOE on live P. acnes-induced inflammatory responses could be due, in part, to the anti-inflammatory and antioxidant properties, but not the anti-microbial effect of EOE.

The excess influx of free fatty acids (FFAs) into nonadipose tissues, such as those of liver and kidney, induces lipotoxicity leading to hepatic steatosis and renal dysfunction. The aim of this study was to investigate the protective effects of methanolic flower extracts of Osmanthus fragrans (OF) and Chrysanthemum morifolium (CM) against FFA-induced lipotoxicity in hepatocytes (human HepG2 cells) and renal glomerular mesangial cells (mouse SV40-Mes13 cells). The results showed that OF and CM significantly suppressed FFA-induced intracellular triacylglycerol accumulation via partially inhibiting the gene expression of sterol regulatory element-binding protein-1c (SREBP-1c) and glycerol-3-phosphate acyltransferase (GPAT) in HepG2 cells. Both extracts inhibited reactive oxygen species (ROS) generation by FFA-stimulated HepG2 cells. OF and CM also suppressed the mRNA expression of interleukin- (IL-) 1β, IL-6, IL-8, tumor necrosis factor- (TNF-) α, and transforming growth factor- (TGF-) β by HepG2 cells treated with conditioned medium derived from lipopolysaccharide-treated THP-1 monocytes. Furthermore, OF and CM effectively inhibited oleate-induced cellular lipid accumulation, TGF-β secretion, and overexpression of fibronectin in mesangial cells. In conclusion, OF and CM possess hepatoprotective activity by inhibiting hepatic fat load and inflammation and renal protection by preventing FFA-induced mesangial extracellular matrix formation.

This study describes algal biofuel production, bioelectricity generation and wastewater treatment using Leptolyngbya sp. JPMTW1 (KF977831). The experiments were conducted in an AMFC (Algal Microbial Fuel Cell) photobioreactor (single-chamber). The pH, EC (electrical conductivity), COD (chemical oxygen demand) and TDS (total dissolved solids) decreased from 8.01 to 7.0, 982 to 854 (mS/cm), 255 to 112 (mg/L) and 490–427 (mg/L), respectively, over course of 7 days. Biomass production, rate of biomass production, chlorophyll a, b and “total chlorophyll” content increased with increasing time and were observed to be 3300 mg/L, 471.42 mg/L/day, 0.981 mg/L, 0.173 mg/L and 1.156 mg/L after 7 days. Lipid production and rate of lipid production were 1068.383 mg/g dry wt. biomass and 152.62 mg/g dry biomass/day. FTIR (Fourier transform infrared) spectra revealed the presence of protein, lipid, FAs (fatty acids), triglycerides and ester functional groups. FAME (fatty acid methyl esters) profile revealed the presence of C16:0, C18:2n-6, C18:1 and C16:1. The generation of electric potential by Leptolyngbya sp. JPMTW1 increased significantly (p ≤ 0.05) from 0.0211 to 0.264 mV within 7 days. The maximum power density (0.008 mW/cm2) was obtained at cell potential at 12 mV. This study shows that simultaneous production of biofuel, bioelectricity and wastewater treatment is possible by Leptolyngbya sp. JPMTW1.

Δ7-Eicosatrienoic acid (Δ7-ETrA; Δ7,11,14–20:3), an elongation metabolite of pinolenic acid (PNA; Δ5,9,12–18:3), is a rare polyunsaturated fatty acid (PUFA) originally from pine seeds. Incorporation of PNA and Δ7-ETrA into murine macrophages inhibited lipopolysaccharide (LPS)-stimulated prostaglandin E2 (PGE2) production. Due to the lack of availability of the naturally-occurring fatty acid, we synthesized Δ7-ETrA and demonstrated it was capable of suppressing PGE2 production. Using laboratory synthetic techniques involving 2-carbon elongation and argentated column chromatography, Δ7-ETrA was synthesized and isolated. Its identity and purity (>98%) were confirmed by gas chromatography (GC)/GC–mass spectroscopy. Incubation of murine RAW264.7 cells or rat primary peritoneal macrophages with Δ7-ETrA reduced PGE2 production by up to 84%, but slightly down-regulated type-2 cyclooxygenase (COX-2) expression. Δ7-ETrA blocked nuclear factor-kappa B (NF-κB) translocation into nucleus and inactivated mitogen-activated protein kinases (MAPK), however, these results might not directly account for its inhibitory effect. Furthermore, PGE2 production reduced by Δ7-ETrA was highly correlated with the extent of Δ7-ETrA incorporation into cellular phospholipids and appeared to be the result of competition between this unusual fatty acid and arachidonic acid (AA) for COX-2. In conclusion, Δ7-ETrA incorporation suppresses PGE2 production by macrophages through competition between Δ7-ETrA and AA for COX-2.

This study investigated the antioxidant and anti-inflammatory properties of the fractionated tea extract, a lower-polymerized polyphenol-rich extract ultrafiltrated from oolong tea. The amounts of total polyphenols, flavonoids and condensed tannin were enriched in fractionated tea extract compared to oolong tea. Furthermore, fractionated tea extract was a stronger scavenger of nitric oxide and 2,2-diphenyl-1-picryhydrazyl radicals than oolong tea. When lipopolysaccharide-treated RAW264.7 macrophages were co-incubated with both tea extracts (500 μg/mL) or (-)-epigallocatechin gallate (100 μg/mL) for 16 h, all samples (oolong tea, fractionated tea extract, and (-)-epigallocatechin gallate) suppressed nitric oxide production by 60, 48, and 46%, respectively, and the suppression was due to nitric oxide-scavenging. However, only fractionated tea extract lowered the proportion of phospholipid arachidonic acid and type II-cyclooxygenase expression, thereby decreasing PGE2 synthesis by 29%. In conclusion, fractio...

Ling-Zhi-8 (LZ-8), a low-molecular weight protein from Ganoderma lucidum, exerts immunomodulatory effects in vitro and in vivo. The aim of this study was to investigate the effects of exposure of LZ-8 to denaturing conditions on its ability to suppress inflammatory responses in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Ultraviolet, fluorescence and circular dichroism spectroscopy analyses showed that LZ-8 was not denatured by exposure to pH extremes, but was irreversibly denatured by temperatures above 55C. LZ-8 lowered the production of nitric oxide and interleukin-6, but had no effect on the level of tumor necrosis factor-α. Heat and acid/alkali-treated preparations of LZ-8 had comparable capacity to suppress proinflammatory mediator production. Since native and denatured LZ-8 were both anti-inflammatory, it appears that this particular biological property of LZ-8 is not dependent on the native structure of the protein, but, possibly on a relatively small peptide domain of LZ-8. Practical Applications Recent evidence suggests that LZ-8, a small protein originally isolated from G. lucidum, has immunomodulatory and anti-carcinogenic properties. In this study, LZ-8 exerted an anti-inflammatory effect on LPS-stimulated macrophages, but this effect was not dependent on the native protein structure, suggesting that the entire protein might not be responsible for the bioactive properties of LZ-8. Since the biological functions of LZ-8 were resistant to heating and exposure to acid or alkali, LZ-8 could have applications in food-processing industries and the production of pharmaceuticals. Furthermore, peptides derived from LZ-8 may be potential targets for development of novel anti-carcinogenic and immunomodulatory agents.

The production of microbial lipids by a batch or fed-batch culture of oleaginous yeast was compared using glucose and corncob hydrolysate as carbon sources. Batch cultures using 4% (w/w) glucose as the carbon source reached a dry biomass, lipid yields and cellular lipid content of 9.4 g/l, 6.0 g/l and 63.5% (w/w), respectively. However, batch cultures demonstrated that there was an inhibitory effect on the biomass and lipid yields when the glucose concentration increased up to 6% (w/w). It was found that the fed-batch culture diminished the substrate’s inhibition of microbial lipid-accumulation. Fed-batch cultures in the corncob hydrolysate-based media containing an initial 4% reducing sugar and an addition 2% glucose for 4 d of cultivation resulted in dry biomass and cellular lipid content of 10.8 g/l and 61.3% (w/w), respectively. The overall lipid productivity increased to 1.08 g/l/d for the fed-batch culture with an initial 4% glucose in corncob hydrolysate and an addition of corncob hydrolysate containing 2% glucose. High concentrations of reducing sugar in the corncob hydrolysate can be efficiently and rapidly converted to Cryptococcus lipid in the fed-batch culture system. Therefore, microbial lipids from Cryptococcus sp. are prospects for a potential alternative oil resource for biodiesel production.

This study investigated the effects of particle-bound polycyclic aromatic hydrocarbons (PAHs) produced from burning three incense types on and their bioreactivity in the RAW 264.7 murine macrophage cell line. Gas chromatography/mass spectrometry was used to determine the levels of 16 identified PAHs. Macrophages were exposed to incense particle extracts at concentrations of 0, 3.125, 6.25, 12.5, 25, 50, and 100 μg/mL for 24 h. After exposure, cell viability and nitric oxide (NO) and inflammatory mediator [tumor necrosis factor (TNF)-α] production of the cells were examined. The mean atomic hydrogen (H) to carbon (C) ratios in the environmentally friendly, binchotan charcoal, and lao shan incenses were 0.69, 1.13, and 1.71, respectively. PAH and total toxic equivalent (TEQ) mass fraction in the incenses ranged from 137.84 to 231.00 and 6.73–26.30 pg/μg, respectively. The exposure of RAW 264.7 macrophages to incense particles significantly increased TNF-α and NO production and reduced cell viability. The cells treated with particles collected from smoldering the environmentally friendly incense produced more NO and TNF-α compared to other incenses. Additionally, the TEQ of fluoranthene (FL), pyrene (Pyr), benzo[a]anthracene (BaA), chrysene (Chr), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1,2,3-cd]pyrene (INP), dibenz[a,h]anthracene (DBA), and benzo[g,h,i]perylene [B(ghi)P] had a significant correlation (R2 = 0.64–0.98, P

Grape-seed procyanidins (GSPs) can inhibit cell proliferation and invasiveness in various human cancers. However, the effect of GSP on pancreatic carcinoma cells has not been investigated.Pancreatic carcinoma cell lines MIA PaCa-2 and BxPC-3 treated with GSP were assessed for viability by trypan blue exclusion, for cell cycle distribution by flow cytometry, for increased apoptosis by annexin V labeling, for their adhesion and invasion potential by evaluating their ability to penetrate through a matrix gel-coated Boyden chamber, and for changes in the levels of proteins involved in cellular events by immunoblotting.Grape-seed procyanidin inhibited MIA PaCa-2 and BxPC-3 proliferation in a dose-dependent manner and induced G1-phase arrest of the cell cycle in BxPC-3 or mitochondria-mediated apoptosis in MIA PaCa-2. Grape-seed procyanidin also inhibited the adhesion and invasion potential of both cell lines in a dose-dependent manner, which are associated with the suppression of metalloproteases matrix metalloproteinase 9 or 2 (MMP-9 or -2) expression.Grape-seed procyanidin inhibited the proliferation of pancreatic carcinoma cells by cell cycle blockage or apoptotic induction. The invasiveness was also suppressed by GSP through down-regulation of MMP-2 or MMP-9 in pancreatic carcinoma cells. Grape-seed procyanidin is a potential chemotherapeutic or preventive agent for pancreatic carcinoma.

Pinolenic acid (PNA), a naturally-occurring polyunsaturated fatty acid (PUFA), is found mainly in pine seeds. Although many studies have demonstrated beneficial effects of pine seed oil, there are no reports of the biological effects of PNA on cancer metastasis. The objective of this study was to investigate the effect of PNA on human breast cancer MDA-MB-231 cell proliferation and metastasis in vitro . We found that PNA did not affect cell viability and cell-matrix adhesion, but it inhibited cell metastasis by suppressing cell invasiveness and motility. Suppression could in part be associated with the modification of the n −6 PUFA composition of cells by PNA which significantly decreased the percentage of arachidonic acid (AA) in phospholipids from 12.6% to 4.9%. The lower AA content of the cancer cells might result in less synthesis of prostaglandin E 2 (PGE 2 ), and subsequent down-regulation of inducible cyclooxygenase (COX-2) expression. Thus, PNA represents a potential anti-cancer agent.

Many reports have shown the beneficial effects of consumption of pine seeds and pine seed oil. However, few studies have examined the biological effect of pinolenic acid (PNA; 5,9,12-18:3), the main fatty acid in pine seed oil. In this study, using murine macrophage RAW264.7 cells as a model, we examined the effect of PNA on polyunsaturated fatty acid (PUFA) metabolism, prostaglandin (PG) biosynthesis and cyclooxygenase-2 (COX-2) expression. Results showed that PNA was readily taken up, incorporated and elongated to form eicosatrienoic acid (ETrA, 7,11,14-20:3) in macrophage cells. A small portion of this elongated metabolite was further elongated to form 9,13,16-22:3. The degree of incorporation of PNA and its metabolites into cellular phospholipids varied with the length of incubation time and the concentration of PNA in the medium. Incubation of PNA also modified the fatty acid profile of phospholipids: the levels of 18- and 20-carbon PUFA were significantly decreased, whereas those of 22-carbon fatty acids increased. This finding suggests that PNA enhances the elongation of 20-carbon fatty acids to 22-carbon fatty acids. The syntheses of PGE(1) from dihomo-gamma-linolenic acid (DGLA, 8,11,14-20:4) and PGE(2) from arachidonic acid (ARA, 5,8,11,14-20:4) were also suppressed by the presence of PNA and its metabolite. As the expression of COX-2 was not suppressed, the inhibitory effect of PNA on PG activity was attributed in part to substrate competition between the PNA metabolite (i.e., 7,11,14-20:3) and DGLA (or ARA).

Sciadonic acid (SCA), pinolenic acid (PNA), and Δ7-eicosatrienoic acid (Δ7-ETrA) are three non-methylene-interrupted fatty acids (NMIFA). Using murine microglial BV-2 cells, this study determined how NMIFA incorporation modulated phospholipid fatty acid composition and the production of pro-inflammatory mediators. Each NMIFA was rapidly taken up and incorporated in BV-2 cells, resulting in the differential redistribution of total lipids. The cellular phospholipid fatty acid compositions were altered, and a significant decrease in the proportions of total monounsaturated fatty acids (MUFA) was observed while the proportions of NMIFA and its metabolites accounted for 38 % of the fatty acid total. Incubation of microglial cells with NMIFA suppressed production of LPS-stimulated pro-inflammatory mediators, including nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), as well as the over-expression of inducible nitric oxide synthase (iNOS) and type 2 cyclooxygenase (COX-2). These inhibitory effects could be accounted for, in part, by the inactivation of mitogen-activated protein kinases (MAPK) signaling. In conclusion, Δ7-ETrA, PNA, and SCA are anti-inflammatory NMIFA that may be useful in suppressing in vitro immune responses involved in neural inflammation.

Propionibacterium acnes is a key pathogen involved in acne inflammation. Wild bitter melon (WBM, Momordica charantia L. var. abbreviate Seringe) is consumed as both a vegetable and as folk medicine in Taiwan. We examined the inhibitory activity of the total phenolic extract (TPE) of WBM leaf on P. acnes-induced inflammatory responses in vivo and in vitro. Our data showed that TPE significantly attenuated P. acnes-induced ear swelling in mice along with microabscess. Flow cytometry analysis revealed that TPE treatment significantly decreased the migration of neutrophils and interleukin (IL)-1β(+) populations in vivo. In P. acnes-stimulated human monocytic THP-1 cells, TPE suppressed the mRNA levels and production of IL-8, IL-1β, and tumor necrosis factor (TNF)-αin vitro. In addition, TPE suppressed P. acnes-induced matrix metalloproteinase-9 levels. TPE blocked nuclear factor-κB (NF-κB) activation and inactivated mitogen-activated protein kinases (MAPK); these actions may partially account for its inhibitory effect on cytokine production. The quantitative HPLC analysis revealed gallic, chlorogenic, caffeic, ferulic, and cinnamic acids, myricetin, quercetin, luteolin, apigenin, and thymol in TPE. All these phenolics significantly suppressed P. acnes-induced IL-8 production in vitro. Our results suggest that WBM leaf extract effectively inhibits P. acnes-induced inflammatory responses and may be useful to relieve the inflammation of acne.

Complementary foods (CF), commonly known as weaning foods, are semi-solid or solid foods that are used to transition infants from breast milk to an adult diet. Their nutritional content is important to the growth and development of children, particularly in developing countries such as Nigeria. In a previous study five CF produced in Jos, Nigeria were analyzed for their nutritional content. Based on those findings, three new CF were formulated in an effort to improve the nutritive value. The new formulations (second-generation CF) were analyzed for fatty acid (FA), amino acid, and mineral and trace element content. The results were compared to those of the most nutritious CF previously analyzed (designated Soy). The total FA content of all three second-generation CF (3.89–20.8 mg/g) was lower than the first-generation Soy mixture (105 mg/g). The content of linoleic and α-linolenic acids among the second-generation CF (1.64–10.1 and 0.084–0.63 mg/g, respectively) was also lower than the Soy CF (59.7 and 7.46 mg/g). The second-generation CF all had higher iron content than Soy (138–288 versus 98.1 μg/g). The amounts of magnesium (1030–1733 versus 2255 μg/g), phosphorus (2237–3830 versus 5685 μg/g), and zinc (28.9–37.9 versus 54.8 μg/g) in the second-generation CF were lower than in Soy. The second-generation CF also had lower protein content than Soy (66–197 versus 355 mg/g). Overall, the new second-generation CF had a lower nutritive content than the original Soy CF.

The precise etiologic factors that initiate and enhance the progression of prostate cancer remains unknown, but diet and lifestyle factors have come forth as significant contributing factors. Conjugated linoleic acid (CLA) is a family of fatty acid isomers found in ruminant food products that appear to have chemopreventive properties in animal models of cancer and antiproliferative effects in several types of cancer cells. Dysregulation and constitutive activation of nuclear factor– κ B (NF κ B) and activator protein 1 (AP1) is found in malignant prostate cells, and dietary components that target these transcription factors may prove to be effective chemopreventive agents. The purpose of this study was to examine the antiproliferative effects of CLA on PC3 cancerous prostate epithelial cells and test the ability of CLA to modulate transcription factors NF κ B and AP1. We found that a mixture of CLA isomers potently inhibited cellular proliferation of androgen-independent PC3 prostate carcinoma cells in a dose-dependent manner. Electromobility shift assays also revealed that CLA inhibited constitutive activation of NF κ B and AP1 transcription factors in PC3 cells. Mobility shift assays demonstrated that CLA also induced activation peroxisome proliferator–activated receptors (PPARs). However, time course studies revealed that PPAR activation occurred after inhibitory effects on NF κ B and AP1, suggesting that inhibition of NF κ B and AP1 did not depend on PPAR activation. The ability of CLA to attenuate inflammatory processes and cellular proliferation by the inhibition of these transcription factors could be the key in their ability to limit cancer development.

Marine microalgae such as Pavlova and Isochrysis produce abundant amounts of the omega3-PUFAs (polyunsaturated fatty acids), EPA (eicosapentaenoic acid, 20:5n-3) and DHA (docosahexaenoic acid, 22:6n-3). The pathway leading to the conversion of EPA into DHA in these lower eukaryotes is not well established although it is predicted to involve an elongation step, catalysed by an elongating enzyme complex, leading to the conversion of EPA into omega3-DPA (omega-3-docosapentaenoic acid, 22:5n-3); followed by a desaturation step, catalysed by a Delta4-desaturase, which results in the conversion of DPA into DHA. To date, the enzymes involved in the elongation of EPA have not been identified from any lower eukaryote. In the present study, we describe the identification of microalgal genes involved in the two-step conversion of EPA into DHA. By expressed sequence tag analysis, a gene (pavELO) encoding a novel elongase was identified from Pavlova, which catalysed the conversion of EPA into omega3-DPA in yeast. Unlike any previously identified elongase from higher or lower eukaryotes, this enzyme displayed unique substrate specificity for both n-6 and n-3 C20-PUFA substrates, with no activity towards any C18- or C22-PUFA substrates. In addition, a novel Delta4-desaturase gene (IgD4) was isolated from Isochrysis, which was capable of converting omega3-DPA into DHA, as well as adrenic acid (22:4n-6) into omega6-DPA. Yeast co-expression studies, with pavELO and IgD4, revealed that these genes were capable of functioning together to carry out the two-step conversion of EPA into DHA.

Dietary supplementation of a high-gamma-linolenic acid canola oil (HGCO) containing approximately 36% (w/w) of gamma-linolenic acid (GLA, 18:3n-6) from the seeds of a genetically transformed canola strain, was assessed for its long-term biological effects. Growing Sprague-Dawley rats (n = 30) were fed a purified AIN93G diet containing 5, 10, or 15% (w/w) of HGCO as the fat source. For comparison, a separate group of rats (n = 10) was given the diet containing 15% (w/w) of borage oil (BO), which contained 22% (w/w) of GLA. After 12 weeks of feeding, the growth, relative organ weights, hematology, and serum biochemistry were found to be similar among rats fed the 5, 10, and 15% HGCO diets. The GLA levels in plasma and liver phospholipids (PL) were also similar. However, the levels of GLA in peripheral tissues (muscle PL and adipose triacylglycerols) were significantly higher in rats fed the 10 and 15% HGCO diets than those fed the 5% HGCO diet. When the above biologic parameters were compared between the 15% HGCO and 15% BO dietary groups, there were no significant differences except for lower final body weights and higher tissue levels of GLA, dihomo-gamma-linolenic acid (20:3n-6) and arachidonic acid (20:4n-6) in the 15% HGCO dietary group as compared with the 15% BO dietary group. This is due to a higher GLA content and possibly a more favorable stereospecific distribution of GLA in HGCO. Overall, long-term (12-week) feeding with diets containing up to 15% HGCO resulted in no adverse effects on growth, organ weight, hematology and serum biochemistry as compared to the diet containing 15% BO, suggesting that HGCO may be a safe alternative source of GLA.

Wild plant foods in the Sahel region of West Africa play an important role in the diets of local residents. During periods of grain shortage, people in rural Niger increase their reliance on wild plant foods to supplement their diets. We report the partial nutrient content of the pit of the seed Sclerocarya birrea , a snack food eaten by children in rural Niger. The pit contained relatively large amounts of copper (24.8 μg/g dry wt), magnesium (4210 μg/g dry wt), and zinc (62.4 μg/g dry wt). The protein content of the pit was high (36.4% of dry wt); however, the protein fraction contained relatively low proportions of leucine, phenylalanine, lysine, and threonine. Fatty acids accounted for 47 mg/g dry wt of the pit, two-thirds of which was due to oleic acid. The essential fatty acid linoleic acid was present (24.5 mg/g dry wt), but the other essential fatty acid, α -linolenic acid, was absent. Such data are useful for health and nutrition program planning by governmental and non-governmental organizations in Niger. The consumption of daniya pits by just children highlights the need to better understand the cultural context of how wild plant foods are used in a particular local context.

Fructose, glucose, and sucrose as the major soluble sugars, and citric, malic, and ascorbic acids as the major organic acids were determined by HPLC, and palmitic acid (16:0) and stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2n-6) and linolenic acid (18:3n-3) as the major fatty acids were determined by GC in medlar (Mespilus germanica 'Dutch') fruit. The compositional changes of these constituents were monitored during the post harvest period of commercially sold Dutch form throughout its rapid ripening (fruit softening-darkening). Sucrose was highest at 1 WAH (228.4 mg/100 g fresh wt) and then decreased, remaining very low at 4 WAH (1.4 mg/100 g fresh wt). As for the levels of fructose and glucose, their levels shifted up to 2230.8 and 845.2 mg/100 g fresh wt at 2 and 3 WAHs, then the levels lessened to their lowest concentration. The levels of the three acids were high at the beginning, except malic acid level at 2 WAH, all acids leveled off through the latter weeks of post harvest period. In the fruit studied, the levels of saturated palmitic acid (16:0) and stearic acid (18:0), and unsaturated oleic acid (18:1), linoleic acid (18:2n-6) and linolenic acid (18:3n-3) were most abundant fatty acids detected throughout medlar ripening (pulp softening and darkening). The level of palmitic and stearic acids as well as the level of linoleic and linolenic acids were the highest at 1 WAH and then suddenly decreased as the medlar soften and the pulp becomes slightly (2 WAH) and fully (3 WAH) brown through 2 and 4 WAHs. In addition to these prominent fatty acids, a remarkable decrease was also obtained in the content of some other fatty acids (C10–15, C16:1, C20–24).

Previous research in rats and mice has suggested that gamma-linolenic acid (GLA) derived from borage oil (BO: 23% GLA) may be an appropriate source for increasing levels of long-chain n-6 FA in the developing brain. Recently, transgenic technology has made available a highly enriched GLA seed oil from the canola plant (HGCO: 36% GLA). The first objective of this study was to compare the effects of diets containing equal levels of GLA (23%) from either BO or HGCO on reproduction, pup development, and pup brain FA composition in mice. The second objective was to compare the effects of the HGCO diluted to 23% GLA (GLA-23) with those of undiluted HGCO containing 36% GLA (GLA-36). The diets were fed to the dams prior to conception and throughout pregnancy and lactation, as well as to the pups after weaning. The behavioral development of the pups was measured 12 d after birth, and anxiety in the adult male offspring was assessed using the plus maze. The findings show that despite equivalent levels of GLA, GLA-23 differed from BO in that it reduced pup body weight and was associated with a slight increase in neonatal pup attrition. However, there were no significant effects on pup behavioral development or on performance in the plus maze. An increase in dietary GLA resulted in an increase in brain 20:4n-6 and 22:4n-6, with a corresponding decrease in 22:6n-3. Again, despite their similar levels of GLA, these effects tended to be larger in GLA-23 than in BO. In comparison with GLA-23, GLA-36 had larger effects on growth and brain FA composition but no differences with respect to effects on reproduction and behavioral development. These findings suggest that the HGCO can be used as an alternative source of GLA.

In mammalian cells, Sprecher has proposed that the synthesis of long-chain PUFA from the 20-carbon substrates involves two consecutive elongation steps, a delta6-desaturation step followed by retroconversion (Sprecher, H., Biochim. Biophys. Acta 1486, 219-231, 2000). We searched the database using the translated sequence of human elongase ELOVL5, whose encoded enzyme elongates monounsaturated and polyunsaturated FA, as a query to identify the enzyme(s) involved in elongation of very long chain PUFA. The database search led to the isolation of two cDNA clones from human and mouse. These clones displayed deduced amino acid sequences that had 56.4 and 58% identity, respectively, to that of ELOVL5. The open reading frame of the human clone (ELOVL2) encodes a 296-amino acid peptide, whereas the mouse clone (Elovl2) encodes a 292-amino acid peptide. Expression of these open reading frames in baker's yeast, Saccharomyces cerevisiae, demonstrated that the encoded proteins were involved in the elongation of both 20- and 22-carbon long-chain PUFA, as determined by the conversion of 20:4n-6 to 22:4n-6, 22:4n-6 to 24:4n-6, 20:5n-3 to 22:5n-3, and 22:5n-3 to 24:5n-3. The elongation activity of the mouse Elovl2 was further demonstrated in the transformed mouse L cells incubated with long-chain (C20- and C22-carbon) n-6 and n-3 PUFA substrates by the significant increase in the levels of 24:4n-6 and 24:5n-3, respectively. This report demonstrates the isolation and identification of two mammalian genes that encode very long chain PUFA specific elongation enzymes in the Sprecher pathway for DHA synthesis.

Five plant-based weaning foods (WF) (Dietrend, Jot-M, Soy, Ang, and Vic-T) locally prepared in Jos, Nigeria were analyzed by gas-liquid chromatography, reverse-phase high performance liquid chromatography, and atomic emission spectrometry with inductively coupled plasma to determine their fatty acid (FA), amino acid, and trace mineral contents, respectively. Results of these direct analyses were compared to expected values derived from food composition tables prepared by the United States Department of Agriculture (USDA). Additionally, results were compared against recommended nutrient values, using breast milk as the standard for FA content and recommended dietary allowances (RDA) for amino acid and mineral contents. The overall nutritional value of the five WF varied considerably and the quantities of particular nutrients determined by direct analysis differed markedly from those estimated using USDA food tables. Comparison of WF fatty acid composition relative to the RDA recommendations and a human milk standard revealed a much higher proportion of both linoleic (35-55 wt%) and alpha-linolenic acids (1%-7 wt%) relative to human milk lipids (11%-12% and 0.8%-0.9% wt, respectively); however, the WF were devoid of arachidonic acid and docosahexaenoic acid. Soy contained the highest amounts of linoleic acid (59.7 mg/g) and alpha-linolenic acid (7.46 mg/g) compared to the other four WF (10.2-41.0 and 0.35-3.18 mg/g, respectively). The linoleic acid/alpha-linolenic acid ratio was within the recommended range (5:1 to 10:1) in only Jot-M (10:1) and Soy (8:1). Dietrend, Vic-T and Ang, contained linoleic/alpha-linolenic ratios of 12:1, 29:1, and 82:1, respectively. The Soy weaning food would provide the most protein (24.3 g/day), based on an estimated daily intake of 65 g of weaning food by a normal six-month-old infant, compared to Jot-M (11.9 g/day), Dietrend (11.7 g/day), Ang (8.07 g/day), and Vic-T (7.26 g/day). The protein RDA for children up to 1 year of age is 13-14 g/day. Comparison of the mineral contents of the WF to the RDAs for various minerals indicated that all five would provide suboptimal amounts of calcium (16 to 250 mg/day) and zinc (1.42 to 3.56 mg/day) compared to respective RDAs of 400 mg/day and 5 mg/day. These data show that the Soy weaning food is an excellent source of linoleic acid and alpha-linolenic acid, as well as being a good source of high quality protein. Jot-M and Dietrend provide useful amounts of the essential FA; however, it is advisable to reevaluate the composition of Ang and Vic-T to find ways to improve the linoleic/alpha-linolenic ratio of each and increase their total protein content. These results document the shortcomings of using published food composition tables based on foods in America when devising weaning foods based on ingredients in another part of the world.

Although abnormalities in the fatty acid composition of serum and red cell membrane phospholipids of patients with type 2 diabetes are well-documented, lacking are studies of this issue in prediabetic individuals.For this cross-sectional study, we recruited 180 subjects (30-80 years), 56 of whom were normal with regard to glucose control (HbA1c,5.7%), 61 who had prediabetes (HbA1c, 5.7%-6.4%) and 59 who had type 2 diabetes (HbA1c,6.5%). Serum phospholipids were isolated and analyzed for fatty acids.Most importantly, the fatty acid compositions of the controls and prediabetic subjects were not different for 19 fatty acids. However, the fatty acid profile of the phospholipids of the patients with diabetes differed from the other two groups; the 14 to 18-carbon saturated fatty acids were decreased by 12%-26% whereas the unsaturated fatty acids 16:1n-7, 18:1n-9, 18:2n-6, 20:3n-6 and 20:4n-6 were increased by 45%-64%. Of note, the docosahexaenoic acid (DHA) status of individuals in all three study groups was remarkably low compared with international values, as indicated by DHA proportions in the 1.62%-2.07% range, and there were no differences between groups. The mean melting point of the phospholipid fatty acids of the diabetic patients (32.2°C) was significantly lower (p0.001) than that of the prediabetic subjects (38.1°C) and the controls (39.9°C) which were not different from each other.These observations indicate that the fatty acid changes associated with type 2 diabetes follow the onset of the disease as opposed to being a causative factor of poor glucose control and insulin insensitivity.

Eicosatrienoic acid (Δ11,14,17-20:3; ETrA) is a rare naturally occurring n-3 polyunsaturated fatty acid (PUFA). Using murine RAW264.7 cells, the objectives were to determine how ETrA modulated phospholipid fatty acid compositions and the production of pro-inflammatory mediators. Incubation cells with ETrA dose-dependently increased the proportions of phospholipid ETrA and its metabolites to 33 % of the fatty acid total. Incorporation of ETrA also reduced the proportions of total n-6 PUFA and monounsaturated fatty acids (MUFA) by 30 and 60 %, respectively. ETrA suppressed LPS-stimulated nuclear factor-kappa B (NF-κB)-mediated nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression. However, no such suppressive effect on the production of prostaglandin E2 (PGE2), cytokines, or expression of cyclooxygenase-2 (COX-2) was observed. As compared with ETrA, eicosapentaenoic acid (EPA) exerted a more potent anti-inflammatory effect. In conclusion, although ETrA suppresses significant NO synthesis and iNOS expression, this n-3 PUFA was a less potent anti-inflammatory agent than EPA.

Background/Purpose : Although docosahexaenoic acid (DHA) is critical for the normal development of the central nervous system, especially in utero and during the 1st year of life, the concentration of this important polyunsaturated fatty acid in the milk fat of Nigerian women is low relative to international standards. There is a compelling need to identify dietary sources of DHA for pregnant and lactating women in sub.Saharan Africa and elsewhere in the world where populations live inland far from the sea and therefore lack access to DHA-rich ocean fish. Henfs eggs represent a potentially useful source of DHA. Methodology : Since the DHA content of eggs varies greatly with the henfs diet and knowing from studies elsewhere in the world that free-range (FR) hens produce eggs enriched for DHA, we compared the fatty acid compositions of the yolk of eggs from FR and gcommercialh hens in northern Nigeria. Results : The percentage of DHA in the FR eggs was 2.6-fold higher (P < 0.001) compared to commercial eggs. Two FR eggs per day could satisfy about three.fourths of the daily recommended intake of DHA for pregnant and lactating women. The FR eggs also contained 2.8-fold (P < 0.001) more α-linolenic acid, more than 10-fold more eicosapentaenoic acid, and 4.1-fold more of the healthful conjugated linoleic acids than commercial eggs. Conclusion : Eggs produced by FR hens could be useful in improving the fatty acid nutrition of pregnant and lactating women in Nigeria, and ultimately their infants who derive much of their nutrition from breast milk. Keywords : Conjugated linoleic acids, docosahexaenoic acid, n.3 polyunsaturated fatty acids, Nigeria, yolk lipids, α-linolenic acid

The seed oil from a genetically transformed canola (Brassica napus) containing 43% (w/w) of γ-linolenic acid (G, 18∶3n−6), 22% linoleic acid (L, 18∶2n−6), and 16% oleic acid (O, 18∶1n−9) was evaluated. In this high γ-linolenic acid canola oil (HGCO), the predominant 18∶3n−6-containing triacylglycerol (TG) molecular species were GGL (23%), GLO (20%), and GGG (11%). In the total TG, approximately 75% of the 18∶3n−6 was located at the sn-1,3 positions, while only 34% of linoleic acid was at the sn-1,3 positions. The GGL molecular species of HGCO contained approximately equal amounts of GLG and GGL positional isomers, while the GLO molecular species had 95% GOL and 5% GLO isomers. The general characteristics and the tocopherol and phytosterol contents were mostly similar between HGCO and nontransformed canola oil. No detectable amounts of amino acids and nucleotides were observed in the HGCO.

Conjugated linoleic acid (CLA; 18:2), a group of positional and geometric isomers of linoleic acid (LA; 18:2n-6), has been shown to modulate immune function through its effect on eicosanoid synthesis. This effect has been attributed to a reduced production of n-6 polyunsaturated fatty acid (PUFA), the precursor of eicosanoids. Since delta6-desaturase is the rate-limiting enzyme of the n-6 PUFA production, it is our hypothesis that CLA, which has similar chemical structure to LA, interacts directly with delta6-desaturase. A unique and simple model, i.e., baker's yeast (Saccharomyces cerevisiae) transformed with fungal delta6-desaturase gene, previously established, was used to investigate the direct effect of CLA on delta6-desaturase. This model allows LA to be converted to y-linolenic acid (GLA; 18:3n-6) but not GLA to its metabolite(s). No metabolites of CLA were found in the lipids of the yeast transformed with delta6-desaturase. The inability to convert CLA to conjugated GLA was not due to the failure of yeast cells to take up the CLA isomers. CLA mixture and individual isomers significantly inhibited the activity of delta6-desaturase of the transformed yeast in vivo. Even though its uptake by the yeast was low, CLA c9,t11 isomer was found to be the most potent inhibitor of the four isomers tested, owing to its high inhibitory effect on delta6-desaturase. Since CLA did not cause significant changes in the level of delta6-desaturase mRNA, the inhibition of GLA production could not be attributed to suppression of delta6-desaturase gene expression at the transcriptional level.

Long-chain polyunsaturated fatty acids (LC-PUFA) are important components of cellular structure and function. Most of LC-PUFA are derived from linoleic acid and alpha-linolenic acid. In plants and fungi, these two acids can be synthesized from oleic acid via the action of two enzymes, delta12 and delta15-desaturases. Due to lack of these enzymatic activities and the ability to synthesize these two essential fatty acids, animals must obtain them from the diet. In this report, we demonstrated the expression of a fungal delta12-desaturase gene in mouse L cells incubated in serum-free medium. The results showed a significant increase in the amount of linoleic acid with a concomitant decrease of oleic acid in cellular lipids. Most of the newly formed linoleic acid was incorporated into cellular phospholipids, particularly phosphatidylcholine. The increase of linoleic acid provided the substrate for the endogenous synthesis of (n-6) LC-PUFA, such as eicosadienoic acid (EDA), dihomo-gamma-linoleic acid (DGLA) and arachidonic acid (AA). Prolonged incubation further increased the levels of linoleic acid derived from oleic acid by the action of delta12-desaturase, and the levels of 20:2n-6 produced from linoleic acid by the action of the endogenous elongase. However, prolonged incubation suppressed significantly the formation of DGLA and AA. In a separate study, a fungal delta6-desaturase gene has also been expressed in the mouse L cells incubated in serum-containing medium. The result shows a significant increase in levels of 20:3n-6 and 20:4n-6. These findings demonstrate that through genetic modification, it is possible to (1) generate cell lines which no longer require dietary 'essential' fatty acids and (2) alter the endogenous fatty acid metabolism to enhance the production of LC-PUFA and their derivatives.

We previously reported that, relative to milk of women elsewhere in the world, the lipid fraction of milk of Fulani women in northern Nigeria contained relatively low proportions of alpha-linolenic acid and docosahexaenoic acid (DHA). This led us to question the essential fatty acid status of Fulani infants and the relation between the proportion of critical n-3 and n-6 fatty acids in the serum phospholipids of the mothers, their milk, and the serum phospholipids of their exclusively breast-fed infants. We were also interested in the effect de novo intermediate chain length-fatty acids (C10-C14) had on the proportions of critical and non-essential fatty acids in milk. Capillary gas-liquid chromatography was used to analyze the fatty acid content of the total milk lipids of 34 Fulani women, as well as the fatty acid content of serum phospholipids of the women and their breast-fed infants during the first 6 months of life. The proportions of critical n-3 and n-6 fatty acids in the milk of the Fulani women were adequate, but the proportions of these same fatty acids were low in their exclusively breast-fed infants. The serum phospholipids of the infants contained 18.8% linoleic acid, 0.13% alpha-linolenic acid, 12.8% arachidonic acid, and 3.40% DHA, whereas, the mean percentages of linoleic, alpha-linolenic, arachidonic and DHA in the serum phospholipids of the Fulani mothers' were 21.4, 0.20, 9.79, and 1.97, respectively. There was a strong positive correlation between fatty acid content of serum phospholipids of Fulani women and the fatty acid content of their milk lipids. As the proportion of C10-C14 fatty acids in the milk lipids increased, the proportions of critical n-3 and n-6 fatty acids in milk remained relatively constant; however, proportions of three non-essential fatty acids decreased dramatically. C10-C14 fatty acids do not appear to displace critical n-3 and n-6 fatty acids in milk.

The Saccharomyces cerevisiae protein ELO2p is involved in the elongation of saturated and monounsaturated fatty acids. Among several sequences with limited identity with the S. cerevisiae ELO2 gene, a consensus cDNA sequence was identified from the LifeSeq(R) database of Incyte Pharmaceuticals, Inc. Human liver cDNA was amplified by PCR using oligonucleotides complementary to the 5' and 3' ends of the putative human cDNA sequence. The resulting full-length sequence, termed HELO1, consisted of 897 bp, which encoded 299 amino acids. However, in contrast with the ELO2 gene, expression of this open reading frame in S. cerevisiae demonstrated that the encoded protein was involved in the elongation of long-chain polyunsaturated fatty acids, as determined by the conversion of gamma-linolenic acid (C(18:3, n-6)) into dihomo-gamma-linolenic acid (C(20:3, n-6)), arachidonic acid (C(20:4, n-6)) into adrenic acid (C(22:4, n-6)), stearidonic acid (C(18:4, n-3)) into eicosatetraenoic acid (C(20:4, n-3)), eicosapentaenoic acid (C(20:5, n-3)) into omega3-docosapentaenoic acid (C(22:5, n-3)) and alpha-linolenic acid (C(18:3, n-3)) into omega3-eicosatrienoic acid (C(20:3, n-3)). The predicted amino acid sequence of the open reading frame had only 29% identity with the yeast ELO2 sequence, contained a single histidine-rich domain and had six transmembrane-spanning regions, as suggested by hydropathy analysis. The tissue expression profile revealed that the HELO1 gene is highly expressed in the adrenal gland and testis. Furthermore, the HELO1 gene is located on chromosome 6, best known for encoding the major histocompatibility complex, which is essential to the human immune response.

Two human expressed sequence tag (EST) cDNA sequences with identity with ∆ 5 - and ∆ 6 -desaturases from a filamentous fungus, Mortierella alpina , were identified from the LifeSeq ® database of Incyte Pharmaceuticals, Inc. (Palo Alto, CA, U.S.A.). An oligonucleotide complementary to the 3ʹ EST cDNA sequences was used to screen human liver cDNA using rapid amplification of cDNA ends (RACE)-PCR. The amplified DNA fragment had 98% identity with a putative open reading frame (ORF) predicted from a human genomic sequence, and encoded 444 amino acids. Expression of this ORF in mouse fibroblast cells demonstrated that the encoded protein was a ∆ 5 -desaturase, as determined by the conversion of dihomo-γ-linolenic acid (C 20:3, n -6 ) into arachidonic acid (C 20:4, n -6 ). The human ∆ 5 -desaturase contained a predicted N-terminal cytochrome b 5 -like domain, as well as three histidine-rich domains. A tissue expression profile revealed that this gene is highly expressed in fetal liver, fetal brain, adult brain and adrenal gland. A search of the existing databases led to localization of this ORF within a 14 kb interval flanked by the flap endonuclease-1 (FEN1) and vitelliform macular dystrophy (Best9s disease; VMD2) loci of chromosome 11q12.

Background Propionibacterium acnes ( P. acnes ) is a commensal bacterium which is possibly involved in acne inflammation. The saturated fatty acid, lauric acid (C12:0) has been shown to possess antibacterial and anti-inflammatory properties against P. acnes . Little is known concerning the potential effects of its decanoic counterpart, capric acid (C10:0). Objective To examine the antibacterial and anti-inflammatory activities of capric acid against P. acnes and to investigate the mechanism of the anti-inflammatory action. Methods The antimicrobial activity of fatty acids was detected using the broth dilution method. An evaluation of P. acnes -induced ear edema in mice was conducted to evaluate the in vivo anti-inflammatory effect. To elucidate the in vitro anti-inflammatory effect, human SZ95 sebocytes and monocytic THP-1 cells were treated with P. acnes alone or in the presence of a fatty acid. The mRNA levels and secretion of pro-inflammatory cytokines were measured by qRT-PCR and enzyme immunoassay, respectively. NF-κB activation and MAPK expression were analyzed by ELISA and Western blot, respectively. Results Lauric acid had stronger antimicrobial activity against P. acnes than capric acid in vitro and in vivo . However, both fatty acids attenuated P. acnes -induced ear swelling in mice along with microabscess and significantly reduced interleukin (IL)-6 and CXCL8 (also known as IL-8) production in P. acnes -stimulated SZ95 sebocytes. P. acnes -induced mRNA levels and secretion of IL-8 and TNF-α in THP-1 cells were suppressed by both fatty acids, which inhibited NF-κB activation and the phosphorylation of MAP kinases. Conclusion Our data demonstrate that both capric acid and lauric acid exert bactericidal and anti-inflammatory activities against P. acnes . The anti-inflammatory effect may partially occur through the inhibition of NF-κB activation and the phosphorylation of MAP kinases.

Propionibacterium acnes is a key pathogen involved in the progression of acne inflammation. The development of a new agent possessing antimicrobial and anti-inflammatory activity against P. acnes is therefore of interest. In this study, we investigated the inhibitory effect of rosemary (Rosmarinus officinalis) extract on P. acnes–induced inflammation in vitro and in vivo. The results showed that ethanolic rosemary extract (ERE) significantly suppressed the secretion and mRNA expression of proinflammatory cytokines, including interleukin (IL)-8, IL-1β, and tumor necrosis factor-α in P. acnes–stimulated monocytic THP-1 cells. In an in vivo mouse model, concomitant intradermal injection of ERE attenuated the P. acnes–induced ear swelling and granulomatous inflammation. Since ERE suppressed the P. acnes–induced nuclear factor kappa-B (NF-κB) activation and mRNA expression of Toll-like receptor (TLR) 2, the suppressive effect of ERE might be due, at least partially, to diminished NF-κB activation and TLR2-mediated signaling pathways. Furthermore, three major constituents of ERE, carnosol, carnosic acid, and rosmarinic acid, exerted different immumodulatory activities in vitro. In brief, rosmarinic acid significantly suppressed IL-8 production, while the other two compounds inhibited IL-1β production. Further study is needed to explore the role of bioactive compounds of rosemary in mitigation of P. acnes–induced inflammation.

Microalgae are good crops to produce natural pigments because of their high growth rates. Tropical zones are better locations than temperate areas for microalgal cultivation because they have longer duration of daylight and more stable temperatures throughout the year, but the high temperatures pose a challenge to microalgal cultivation. A newly isolated thermotolerant microalga produces reddish pigments under environmental stress. Morphological and molecular evidence including meridional ribs on the cell wall, pigment production, and its 18S rDNA sequence suggests that this microalga belongs to the genus Coelastrella. Salt stress and high light intensity accelerated biosynthesis of the pigments, and significant quantities of oil accumulated as the cells experienced stress due to nutrient deficiency. This microalga could withstand temperature of 50°C for more than 8h, which is a necessary trait for outdoor cultivation in tropical areas. The pigments contain astaxanthin, lutein, canthaxanthin, and β-carotene as analysed by using HPLC.

Sciadonic acid (SCA; Δ5,11,14-20:3), a non-methylene-interrupted polyunsaturated fatty acid (NMIFA), can substitute for arachidonic acid (AA) and reduce prostaglandin E2 (PGE2) synthesis in macrophages. However, little is known about how SCA exerts its anti-inflammatory effects. The objectives of this study were to purify SCA from seeds of Podocarpus nagi and investigate mechanisms underlying the modulatory effects of SCA on inflammatory responses in murine RAW264.7 macrophages. We describe how high-purity SCA (>98%) can be obtained using argentated column chromatography. SCA was dose-dependently incorporated into cellular phospholipids, and increasing SCA incorporation correlated with decreases in the proportions of AA, total monounsaturated fatty acids (MUFA) and total saturated fatty acids (SFA). SCA decreased production of PGE2 (29%), nitric oxide (NO) (31%), interleukin-6 (IL-6) (34%) and tumor necrosis factor-α (TNF-α) (14%). The suppression of pro-inflammatory mediators was due, in part, to decreased expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). SCA incorporation suppressed nuclear factor-kappa B (NF-κB) translocation and phosphorylation of mitogen-activated protein kinases (MAPK), including extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK). These findings indicate that by altering the cellular fatty acid composition SCA can modulate the responsiveness of macrophages to LPS through inactivation of NF-κB and MAPK pathways.