Three experiments were conducted to investigate the hypothesis that cows grazing on pasture produce the highest proportion of c-9 t-11 CLA in milk fat and no further increase can be achieved through supplementation of diets rich in linoleic acid, such as full-fat extruded soybeans or soybean oil. In experiment 1, 18 lactating Holstein cows were used in a randomized complete block design with measurements made from wk 4 to 6 of the experiment. In experiment 2, three cannulated lactating Holstein cows were used in a $3{\times}3$ Latin square design. Each period was 4 wk with measurements made in the final wk of each period. Cows in both experiments were assigned at random to treatments: a, conventional total mixed ration (TMR); b, pasture (PS); or c, PS supplemented with 2.5 kg/cow per day of full-fat extruded soybeans (PES). In both experiments, feed intake, milk yield, milk composition, and fatty acid profile of milk and blood serum were measured, along with fatty acid composition of bacteria harvested from rumen digesta in experiment 2. In experiment 3, 10 cows which had continuously grazed a pasture for six weeks were assigned to two groups, with one group (n = 5) on pasture diet alone (PS) and the other group (n = 5) supplemented with 452 g of soy oil/cow per day for 7 d (OIL). In experiment 1, cows in PS treatment produced 350% more c-9, t-11 CLA compared with cows in TMR treatment (1.70 vs. 0.5% of fat), with no further increase for cows in PES treatment (1.50% of fat). Serum c-9, t-11 CLA increased by 233% in PS treatment compared with TMR treatment (0.21 vs. 0.09% of fat) with no further increase for cows in PES treatment (0.18% of fat). In experiment 2, cows in PS treatment produced 300% more c-9 t-11 CLA in their milk fat compared with cows in TMR treatment (1.77 vs. 0.59% of fat), but no further increase for cows in PES treatment (1.84% of fat) was observed. Serum c-9, t-11 CLA increased by 250% for cows in PS treatment compared with cows in TMR treatment (0.27 vs. 0.11% of fat), with no further increase for cows in PES treatment (0.31% of fat). The c-9, t-11 CLA content of ruminal bacteria for cows in PS treatment was 200% or more of TMR treatment, but no further increase in bacterial c-9, t-11 CLA for cows in PES treatment was observed. Supplementation of soy oil in experiment 3 also did not increase the c-9 t-11 CLA content of milk fat compared with cows fed a full pasture diet (1.60 vs. 1.54% of fat). Based on these findings, it was concluded that supplementing with feeds rich in linoleic acid, such as full-fat extruded soybeans or an equivalent amount of soy oil, to cows grazing perennial ryegrass pasture may not increase milk fat c-9 t-11 CLA contents.
Kim, Jung-Hyun;Hubbard, Neil E.;Lim, Debora;Erickson, Kent L.
Preventive Nutrition and Food Science
/
v.11
no.1
/
pp.1-5
/
2006
Conjugated linoleic acid (CLA) is a group of positional and geometric isomers of linoleic acid that have been used to reduce the incidence, growth and metastasis of breast, colon, prostate and gastric cancer in animals. CLA could reduce tumor growth by altering angiogenesis; a process requiring associated angiogenic factors such as vascular endothelial growth factor (VEGF). In this study, we determined whether CLA could modulate the expression of VEGF in murine mammary tumor cells and adipocytes. The c9, t11-CLA isomer reduced VEGF transcripts and protein when mammary tumor cells were stimulated with PMA. That isomer also reduced VEGF expression in un stimulated mouse 3T3-L1 adipocytes. Since VEGF can be regulated by cyclooxygenase-2 (COX-2), we determined whether CLA could alter COX-2 enzyme expression and $PGE_2$ production. The c9, t11-CLA isomer reduced not only COX-2 enzyme expression but also $PGE_2$ production. Thus, c9, t11-CLA could modulate neovascularization by alteration of VEGF expression from mammary tumor cells and adipocytes by reducing COX-2 metabolites.
Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.
Dietary conjugated linoleic acid (CLA) play key roles in lipid metabolism. Here, we investigated the effect of CLA on the transcriptional activity of TR4, an orphan nuclear receptor that plays an important role in lipid homeostasis. CLA increased TR4 gene mRNA level in 3T3-L1 adipocytes, but inhibited TR4 transcriptional activity in a dose-dependent manner. TR4 induced perilipin expression in 3T3-L1 adipocytes by activating perilipin promoter activity. In a gel shift assay, TR4 bound direct to the putative TR4 response element in the perilipin promoter. Interestingly, CLA reduced the interaction between TR4 and consensus DR1, a well-known TR4 binding site. Additionally, CLA inhibited TR4-induced perilipin promoter activity in a dose-dependent manner. Together, our results suggest that CLA may play a role in lipid homeostasis in adipocytes by functionally regulating TR4.
This study was conducted to investigate the effects of Cordyceps ochraceostromat, silkworm cocoon, and conjugated linoleic acid (CLA) on the quality and storage properties of pork sausage manufactured with protein recovered from breast of spent laying hen during 4 wks of storage at $4^{\circ}C$. Pork sausages were prepared using 100% ham (control) and 40% recovered protein from breast of spent laying hen to replace pork (T1), and with added different sources to final concentrations of 0.1% Cordyceps ochraceostromat powder (T2), 0.1% silkworm cocoon powder (T3), 0.1% CLA (T4), 0.05% Cordyceps ochraceostromat + 0.05% silkworm cocoon (T5), 0.05% Cordyceps ochraceostromat + 0.05% CLA (T6), and 0.05% silkworm cocoon + 0.05% CLA (T7). The treatments T5 and T7 had higher (p<0.05) protein content than control, but control had lower fat content than other samples during 4 wks of storage at $4^{\circ}C$. Lightness was significantly lower in the treatment samples than control. However, there was no significant difference in water holding capacity between the sausage samples, whereas, cohesiveness and chewiness were significantly higher (p<0.05) in the control than other treatments. All sausage samples showed a significant increase in volatile basic nitrogen (VBN) and total plate counts with extending storage time (p<0.05), and VBN values of treatments were lower than the control. However, the treatment samples showed a significant decrease (p<0.05) in thiobarbituric acid reactive substances over the increasing storage time. Therefore, our results suggested that the 40% recovered protein to replace pork and with added different sources decreased lipid oxidation and protein denaturation of pork sausages, thereby enhancing self-life, compared to normal pork sausage (control).
Park, So-Young;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
Journal of Veterinary Clinics
/
v.31
no.6
/
pp.469-476
/
2014
The aims of this study were to explore the effects of conjugated linoleic acid (CLA) on reactive oxygen species (ROS) production in lipopolysaccharide (LPS)-naïve and LPS-stimulated RAW 264.7 macrophages and to examine whether these effects affect the regulation of tumor necrosis factor-alpha (TNF-${\alpha}$) production, and nuclear factor-kappa B (NF-${\kappa}B$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) activation. Trans-10, cis-12(t10c12)-CLA increased the production of ROS, as well as TNF-${\alpha}$ in LPS-naïve RAW 264.7 cells. The CLA-induced TNF-${\alpha}$ production was suppressed by treatment of diphenyleneiodonium chloride (DPI), a NADPH oxidase inhibitor. In addition, CLA enhanced the activities of NF-${\kappa}B$ and $PPAR{\gamma}$ in LPS-naïve RAW 264.7 cells, and this effect was abolished with DPI treatment. LPS treatment increased ROS production, whereas CLA reduced LPS-induced ROS production. LPS increased both TNF-${\alpha}$ production and NF-${\kappa}B$ activity, whereas t10c12-CLA reduced TNF-${\alpha}$ production and NF-${\kappa}B$ activity in LPS-stimulated RAW 264.7 cells. DPI treatment suppressed LPS-induced ROS production and NF-${\kappa}B$ activity. Moreover, DPI enhanced the inhibitory effects of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ activation in LPS-stimulated RAW 264.7 cells. However, neither t10c12-CLA nor DPI affected $PPAR{\gamma}$ activity in LPS-stimulated RAW 264.7 cells. Taken together, these data indicate that t10c12-CLA induces TNF-${\alpha}$ production by increasing ROS production in LPS-naïve RAW 264.7 cells, which is mediated by the enhancement of NF-${\kappa}B$ activity via $PPAR{\gamma}$ activation. By contrast, t10c12-CLA suppresses TNF-${\alpha}$ production by inhibiting ROS production and NF-${\kappa}B$ activation via a $PPAR{\gamma}$-independent pathway in LPS-stimulated RAW 264.7 cells. These results suggest that t10c12-CLA can modulate TNF-${\alpha}$ production and NF-${\kappa}B$ activation through formation of ROS in RAW 264.7 macrophages.
Conjugated linoleic acid (CLA) isomers are found naturally in foods, such as milk, milk products, beef and others, from biohydrogenation of vegetable oils. They are heterogenous group of isomers of linoleic acid in the family of polyunsaturated fatty acids. Among the isomers of linoleic acid cis9, trans11- CLA (c9, t11-CLA) and trans10, cis12- CLA (t10, c12-CLA) are found to be biologically active isomers. These biologically active isomers either individual or combined found to be health beneficial in various diseases, such as cancer, diabetes, obesity, and atherosclerosis, conclusive participation in physiological processes are necessary. This review focused on the current study of CLA in prevention of disease, such as cancer, diabetes and atherosclerosis, and their effective function in body fat reduction, improvement of bone and muscle mass at a cellular, clinical and systematic level.
Iga, T.;Satoh, T.;Yamamoto, S.;Fukui, K.;Song, S.H.;Choi, K.C.;Roh, S.G.;Sasaki, S.
Asian-Australasian Journal of Animal Sciences
/
v.22
no.11
/
pp.1566-1573
/
2009
Trans-10, cis-12 conjugated linoleic acid (CLA) has been reported to inhibit the adipocyte differentiation of preadipocytes in non-ruminant animals (mice, rat, and human). However, the effects of trans-10, cis-12 CLA have not been clear in ruminants. The objective of this study was to investigate the effects of trans-10, cis-12 CLA on adipocyte differentiation of ovine preadipocytes. Differentiation of these preadipocytes was facilitated by treatment with trans-10, cis-12 CLA. Trans-10, cis-12 CLA increased the number and size of oil red O-stainable lipid drops as well as the levels of GPDH activity. PPAR-$\gamma{2}$ and adipophilin mRNA, adipogenic marker genes, were increased by treatment with trans-10, cis-12 CLA. This result was different from that observed with 3T3-L1 preadipocytes, a clonal cell line derived from rodents. Furthermore, trans-10, cis-12 CLA alone induced the adipocyte differentiation of ovine preadipocytes in differentiation-induction medium without troglitazone. These results suggest that CLA is an inducer and regulator in adipocyte differentiation of ovine preadipocytes, with species differences between ovine and rodent preadipocytes.
Variations in conjugated linoleic acid (CLA) concentrations in Holstein dairy cows milk, depending on feeding systems in different seasons was investigated. Milk samples were collected from Holstein dairy cows, which either grazed for whole days (WG), only daylight hours (TG), or were offered a total mixed ration (TMR) and experienced no grazing (NG), from April to December of 2005. In April, November and December, the cows in TG and WG treatments received grass silage and some concentrate, while from May to October, the cows grazed on temperate pasture. The cows in NG treatment received the TMR throughout the season. The major fatty acid obtained in the pastures was linolenic acid. There was no significant difference in the pasture's linolenic acid concentrations from May to September, but there was a significant decrease in October. However, the linolenic acid concentrations obtained in the pasture were always much higher than those obtained from the TMR. Linoleic acid was also the major fatty acid in the TMR, but these concentrations were higher in the TMR than in the pasture. There was no significant difference in milk cis9trans11CLA (c9t11CLA) concentrations between the three feeding systems while the cows were fed on conserved pasture in April, November and December. Although c9t11CLA concentrations were lower in the TMR, it was found that the cows which grazed in fresh pasture experienced significantly higher concentrations of c9t11CLA in their milk than those which received only TMR. It was also found that cows in the WG treatment experienced higher c9t11CLA concentrations than those in the TG treatment. In the WG and TG treatments, c9t11CLA concentrations were highest in June, after which, they gradually decreased (p<0.01) until October. For the NG treatment, there was no significant change in the concentrations of c9t11CLA (p>0.05) with season. Overall, trans11C18:1 and c9t11CLA were greatly influenced by season, with higher variation in the WG treatment than in the TG treatment and no variation in the NG treatment.
Kim, Young-Beum;Lee, Ill-Woo;Kang, Ji-Houn;Yang, Mban-Pyo
Journal of Veterinary Clinics
/
v.28
no.2
/
pp.190-195
/
2011
Nuclear factor ${\kappa}B$ (NF-${\kappa}B$) is a nuclear transcription factor that modulates the expression of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. trans-10, cis-12 (t10c12)-conjugated linoleic acid (CLA) participates in the inhibition of TNF-${\alpha}$ production upon lipopolysaccharide (LPS)-stimulation. However, in our previous study, t10c12-CLA enhanced the production of TNF-${\alpha}$ by LPS-unstimulated porcine peripheral blood mononuclear cells (PBMCs) and RAW 264.7 macrophages in vitro. To resolve this apparent contradiction, we hypothesized that the effect of t10c12-CLA on TNF-${\alpha}$ production depends on NF-${\kappa}B$ activation induced by LPS stimulation. To test this hypothesis, we assessed the in vitro effect of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ p65 activity in LPS-stimulated and LPS-unstimulated porcine PBMCs. t10c12-CLA treatment resulted in increased TNF-${\alpha}$ production by LPS-unstimulated PBMCs but decreased TNF-${\alpha}$ production by LPS-stimulated PBMCs. t10c12-CLA increased the degradation of inhibitory ${\kappa}B$ ($I{\kappa}B$)-${\alpha}$ protein and activated NF-${\kappa}B$ p65 in LPS-unstimulated PBMCs, but had the opposite effect in LPS-stimulated PBMCs. Notably, t10c12-CLA enhanced NF-${\kappa}B$ p65 binding activity in LPS-unstimulated PBMCs exposed to caffeic acid phenethyl ester (CAPE), a NF-${\kappa}B$ inhibitor. Conversely, it inhibited NF-${\kappa}B$ p65 binding activity in LPS-stimulated PBMCs exposed to CAPE. These results suggest that t10c12-CLA may have different actions under different physiological conditions, and that its effect may be associated with a change in NF-${\kappa}B$ p65 activity.
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