Eur. J. Lipid Sci. Technol.

2011, 113, 1077–1094

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Review Article Long-term conjugated linoleic acid supplementation in humans – effects on body composition and safety
Remo P. Jutzeler van Wijlen
¨ Sponser AG, Furtistrasse 5, CH-8832 Wollerau, Switzerland

In our contemporary adipogenic environment even modest improvements in body fat mass could be of relevance. In the last years animal and human studies have investigated the potential benefit of CLA on body composition. However, inconclusive results are often derived from short-term studies. Long-term intervention trials with supplemental CLA on body composition have not been reviewed exclusively up to now. Therefore, the objective of this study was to review the evidence of prolonged CLA supplementation as well as its influence on body composition in humans, and to summarize results from safety assessments of CLA intake. A literature search was performed to find intervention trials with CLA supplementation and its effects on body composition, as well as on insulin sensitivity. Only prolonged (!12 wk) studies on body composition were included. The investigated studies indicate a modest reduction and/or prevention of regain of body fat in overweight/obese subjects. Results on the influence of CLA on insulin sensitivity are inconsistent, with newer data rather adding to the safety of CLA. Impaired insulin sensitivity by CLA remains a safety concern, yet is seemingly restricted to diabetic subjects and single-isomer application. A meta-analysis of extended studies only is warranted to quantitatively evaluate the effects of CLA on body composition. Future research may elucidate if CLA should be considered as a marginal missing, semiessential nutrient in our present diet.
Keywords: Body composition / CLA / Insulin / Review / Safety

Received: March 31, 2011 / Revised: May 9, 2011 / Accepted: May 23, 2011 DOI: 10.1002/ejlt.201100130

1 Introduction
CLA refers to a group of PUFA with geometrical and positional isomers with conjugated double bonds (from 6.8 to 12.14 in cis/cis, trans/cis, cis/trans, trans/trans configuration) [1]. The multiple-step biohydrogenation of linoleic acid to c9,t11 octadecadienoic acid was discovered already in 1966 by Kepler et al. [2], though the term CLA was not established yet. In 1978, the unexpected anti-carcinogenic activity found in hamburgers by Pariza et al. [3] was assigned to CLA during subsequent work [4]. In 1998, the predominant c9,t11 isomer (>80%) of naturally occurring CLA in food

Correspondence: Dr. Remo P Jutzeler van Wijlen, Sponser AG, CH-8832 . Wollerau, Switzerland E-mail: r.jutzeler@sponser.ch Fax: þ41 43 888 1800 Abbreviation: BF, body fat; BM, body mass; BMI, body mass index; CRP, C-reactive protein; DEXA, dual energy X-ray absorptiometry; FFA, free fatty acids; HOMA-R, homoeostasis model assessment; IL-1b, Interleukin-1b; IL-6,8, interleukin-6,8; IL-10, Interleukin-10; LBM, lean body mass; Lp(a), lipoprotein (a); RQ, respiratory quotient; SFA, saturated fatty acids; TNF-a, tumour necrosis factor-a ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

sources [5, 6] was named rumenic acid (RA), as proposed by Parodi [7], due to its formation in ruminants by microbial hydrogenation of linoleic and linolenic acid. However, the predominant origin of RA is not ruminal [8–10], but from endogenous desaturation of trans fatty acids (TFA) in the adipose tissue and mammary gland [11]. CLA shows a linearly correlated concentration in cow and goat milk with vaccenic acid (VA, C18:1-t11) [12, 13], typically the main trans-C18:1 isomer in milk fat and an important precursor of CLA. Also other CLA isomers are found in considerable amounts (!2% of total fatty acids) in beef and dairy products [14–16]. Dietary CLA availability through dairy products and meat can amount to >1 g/d, if animal-adapted, grassbased feeding strategies are applied and endogenous bioconversion of vaccenic acid to CLA is considered [17, 18]. CLA showed promising results in reducing fat mass and increasing fat-free mass in animal studies (see reviews: [19, 20]). In consequence of the pandemic-like growth of overweight and obesity, the potential benefit of CLA on body composition has gained attention both by popular and scientific publications. In humans, however, the conclusions are ambiguous in many ways. Therefore, the objective of this study was to review the evidence on CLA and body composition in humans. During the last years several long-term
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Eur. J. Lipid Sci. Technol. 2011, 113, 1077–1094

human studies with CLA interventions have been published. For the most part, studies conducted in animals as well as in humans have been done by using synthetic mixtures of isomers (mainly c9,t11 and t10,c12 at a %50:50 ratio), along with concomitant minor amounts of other isomers. This fact is also reflected in present CLA supplements available on the market, which are solely the 50:50 ratio mixture. In addition to evaluate the significance and relevance of any effect on body composition, it is important to look for possible negative side-effects of CLA. For example, in a few clinical trials CLA has been shown to induce inflammation, lipid peroxidation and influence negatively insulin sensitivity [21, 22], though seemingly these effects are restricted to the isolated t10,c12 single-isomer and short-term CLA supplementation [23, 24]. Therefore, safety aspects related to CLA supplementation are discussed. Other health aspects of longor short-term CLA supplementation are not covered in this overview. However a wealth of literature of a more general nature [25–28] as well as specific studies on CLA and its modulating effects on blood lipid profile [29, 30], insulin sensitivity and diabetes [21, 31–37], atherosclerosis [38–40], anti- and pro-oxidant activity [21, 22, 41, 42], anti-inflammatory action [43–45], carcinogenesis [46–50], bone formation and maintenance [51–54] and immune system [55–57] are available for the interested reader.

adipose tissue [61–63] and in breast milk [64, 65] of humans. The c9,t11 isomer is seemingly the only one present in adipose tissue [66]. CLA are incorporated in both TAGs and phospholipids [67]. In the latter CLA out-compete AA at the same molecular position, though sole exchange does not explain for all the reduction of AA levels as stated previously [68]. But in that review it is pointed at the reported inhibiting effect of CLA on cyclooxygenase activities, the rate limiting enzyme for prostaglandin formation [69].

3.1.2 Mechanisms of action in CLA-mediated body composition changes
Animal and human studies suggest different mechanisms how CLA affects body composition [70, 71]. These mechanisms include increased basal metabolic rate [72, 73], decreased respiratory quotient and consequently increased fat oxidation [74], inhibition of stearoyl-CoA desaturase activity [75], inhibition of lipoprotein lipase (LPL) activity [76], induction of adipocyte apoptosis [77] and/or adipocyte differentiation [25, 78]. Other hypothesized mechanisms are preadipocyte proliferation, reduced TAG accumulation and modulation of de novo adipose tissue lipid synthesis via influencing adipogenic gene expression mechanisms (including PPAR-g, NFkB, TNF-a), as well as leptin levels [79–81]. Some mechanisms and effects of CLA regarding lipid metabolism are postulated to be similar to other poly-unsaturated fatty acids (PUFA) [82], such as, e.g. the impact on the neuroendocrine system via influencing leptin and adiponectin levels [83]. Recent evidence showed an increase in nocturnal fat oxidation as well as reduced carbohydrate oxidation, resulting in a lower respiratory quotient (DRQ: À0.01 vs. 0.02, p 0.05) after 6 months of 3.2 g/d CLA supplementation [74]. Additionally, protein utilization during sleeping hours in percentage of energy was lower with CLA (D%E À3.3 vs. 0.3, p < 0.05). In total, this suggests higher fat oxidation and a protein-sparing effect during the night. Since neither dietary fat oxidation during daytime (only positive trend) nor 24 hmacronutrient utilization were increased, an elevated endogenous or dietary fat oxidation during sleeping hours was postulated [74]. Overall, it is still unclear which ones of these multiplesteps mechanisms involving CLA are causal regarding the modulation of body composition, and which ones are effects. However, it seems that the CLA related changes in fat accumulation and lipid metabolism are closer linked to the t10,c12 than the c9,t11 isomer [77, 84, 85]. But it is deemed important to keep in mind gender-related differences in lipid metabolism, potentially leading to different regional fat distribution between genders [86]. It is also postulated that phenotypic traits (e.g., body composition, regional fat distribution) may cause differences in lipid kinetics, but itself could also be the result of sexual dimorphism in respect to
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2 Literature data and selection criteria
Data for this review were obtained by searching the regularly updated, web-based reference listing on CLA [58], which has been keeping track of published studies on CLA since at least 1987, the year of coinage of the term CLA. The reference listing was searched against certain inclusion criteria for references published during the past decade. The criteria were: studies and reviews in humans (i) with CLA on body composition and a minimum of 12 wk supplementation length, (ii) on safety and (iii) on biomechanisms. Reference lists of retrieved studies were also scanned to ensure completeness. Studies on body composition in animals, as well as in vitro or ex vivo studies have been disregarded. In addition, for completion reasons and to uncover older studies of interest on human CLA intake, a search by term (CLA, conjugated linoleic, humans, body composition, insulin, intake, content, review) was undertaken using PubMed [59] and Google scholar [60]. The search considered studies published until the end of April 2011.

3 Results and discussion
3.1 Influence of CLA on body composition 3.1.1 CLA incorporation in humans
Dietary intake of CLA correlates with levels in plasma as well as in cellular lipids and also translates into CLA content in
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lipid metabolism [87]. Furthermore, dietary intervention with TFA revealed gender-specific differences in gene expression [88].

3.1.3 Human studies on modulation of body composition
Out of the studies referring to CLA and body composition only those with a minimum length of 12 wk supplementation were considered for reasons discussed hereafter. A sustainable body mass (BM) management implies compliance over an extended period of time, and any dietary supplement or food ingredient to support crash diet procedures seem inappropriate. The potential inefficacy or even deleterious outcome of such regimens are described in the literature [89, 90]. Eventually, 24 studies on modulation of body composition with appropriate study length were identified and included in this review (Table 1). Sixteen out of them were conducted on a prolonged (!12 wk) and seven on a long-term base (!6 months), whereof one was extended as an open follow-up for another 12 months. Six other retrieved studies of appropriate study length were disregarded in the context of modulation of body composition because primarily designed to measure the influence of CLA on glucose and insulin [91], combined with a very low number of only nine subjects [92], a combined supplementation of CLA with n3-fatty acids [93], amino acids [94], or chromium picolinate [95], or a design considered not precise enough to measure changes of body composition [96]. A significant association of supplemental CLA and body composition was reached in some aspects, e.g. BM, body mass index (BMI), body fat (BF), waist circumference, or lean body mass (LBM), in the majority [31, 97–110] but not in all [23, 37, 73, 111–114] reviewed studies (Table 1). Yet, of these latter ones, significant changes regarding BM and fat mass have been demonstrated within case groups, but not compared to placebo in two studies [113]. Another study observed modest, but not significant fat loss rather peripheral than central [114], and again another found an increased basal energy expenditure (D: À1.2 vs. þ4.6 kJ/kg/d fat mass, p ¼ 0.03) [73]. Data of other studies were not available [102, 115], or study duration was only short-term [116–121]. It is difficult to evaluate the inconclusive outcomes because design, subjects and anthropometric measurement methods were strongly varying study characteristics, whereas the used CLA isomers and doses have been similar in most trials. Supplemental CLA was usually given as a 50:50 isomeric mixture of c9,t11 and t10,c12 with a dosage between 3.0 and 6.4 g/d, the rationale behind being specific isomer effects. Median daily dosage of the reviewed studies was 3.5 g. Two reviews reported no effect of CLA on body composition [122, 123]. However, both of them included also shortterm studies. A modest improvement in body composition was demonstrated in the first meta-analysis of 18 human studies (though three of them were with a single-isomer
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design) on CLA supplementation [124]. According to the authors, the ten of the analysed studies, which showed no significant effect on fat mass, lacked statistical power because there were too few subjects, treatment was too short, or both. Nevertheless, the conclusion of that meta-analysis was a modest, but highly significant average fat loss of 0.09 kg per wk ( p < 0.001) compared to placebo. Their argumentation states that an expected fat loss after 12 wk would have been 1.1 kg, but the SD for within-individual change was 2.6 kg, necessitating 44 participants in each group to have 80% power to detect this change with p < 0.05 [124]. Furthermore, theoretical calculations indicate that a reduction in BF caused by potentially increased energy expenditure after CLA supplementation would be about seven times lower in humans than in mice because of a seven-fold higher murine metabolic rate per kg BM [72]. Therefore, in spite of an observed, CLA-induced, higher basal energy expenditure [73] and lower respiratory quotient [74], duration of supplementation seems a crucial factor and, if too short, may have been decisive for a lack of effect on body composition in many studies. Already previously weight loss was not detected until after 8 wk of CLA supplementation [108]. Importantly, animal studies focussed on reduced accretion of BF in growing subjects, in contrast to the examination of reducing existing BF in adult, i.e. non-growing humans. Only recently human studies have been looking at BM and fat regain rather than depot fat reduction, and showed promising results [98, 111, 125]. Similarly, an interesting piglet model simulating weight gain during human infancy found reduced BF gain and lipogenic gene expression by supplementing 1% of dietary energy (%E) from CLA in both a low-fat (3%E) and a high-fat (25%E) diet [126]. A first placebo-controlled trial on prepubescent children supports this observation: a reduced fat mass accretion ( p ¼ 0.001) and attenuated BMI increase ( p ¼ 0.05) was measured after 7 months of 2.4 g CLA supplementation [109]. A common parameter in virtually all studies on CLA and body composition, quite reasonable, is that subjects were overweight to obese (BMI 25þ). Only one long-term [127] and one recent, though short-term [121], study worked with subjects with a BMI < 25. However, recent data indicate that dietary fatty acid composition may influence gene expression differently in lean and overweight men [128]. Supplementation for 9 days with a dietary fat spread rich in CLA (4.96 g/d) and medium chain TAG caused considerable variation in adipose tissue gene expression, mainly with respect to defence response, energy and lipid metabolism. Although the results indicated a weaker and more variable response in overweight subjects to the intervention spread, it is remarkable that expression of a number of genes involved in lipid metabolism and inflammation, which were significantly different compared to lean subjects at baseline, changed in the direction of expression levels in lean subjects. In the majority of overweight subjects the expression of inflammawww.ejlst.com

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Table 1. Studies on CLA and modulation of body composition with a minimum supplementation length of 12 wk.
Body composition þ/À effect on metabolic factors –

Year ref. Isomers 50:50 c9,t11/ t10,c12 n.i. Safflower oil –À 0 DEXA, skin fold thickness ÀÀ Form Placebo BM LBM BF Anthropometrics 35 postmenopausal diabetic women 35–38.5 6.4 g

Design and approach

Length of supplementation Baseline BMI CLA [g/d]

Study finishersa)

No effect on metabolic parameters vs. placebo Insulin, glucose, adiponectin, leptin, ALT, AST, HOMA-R

R. P Jutzeler van Wijlen .

2010 [108]

Cross-over design (2 x 16 w), body composition, markers of hepatic and glycemic function, no diet or lifestyle restriction 53 prepubertal overweight to obese children !85th age %ile 2.4 g 50:50 c9,t11/ t10,c12 TAG Sunflower oil DEXA ÀÀ þþ ÀÀ Reduced HDL, bone mineral accretion

16 wk

ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Insulin, glucose, LDLand total cholesterol, TAG, ALT, AST, HOMA-R 75 postmenopausal women 22.4–28.8 5.5 50:50 c9,t11/ t10,c12 TAG Olive oil ÀÀ þþ (lower-body) ÀÀ DEXA reduced GLUT4, LPL, leptin, increased TNF-a insulin, leptin, adiponectin, glucose, HOMA-R 83 28–32 3.4 50:50 c9,t11/ t10,c12 FFA Olive oil À Àb) þþy ÀÀ DEXA Reduced HDL, increased CRP and insulin c-peptide (all within normal range), insulin and glucose reduction Lp(a), LDL- and total cholesterol, TAG, leucocytes, IL-6, IL-8, TNF-a, leptin, adiponectin, alanine-amino transferase, alkaline phosphatase, creatinine, erythrocytes, thrombocytes ÀÀ 0 À 4-compartment model, DEXA Decreased abdominal fat (þ), decreased sVCAM-1 (decreased endothelial dysfunction) TAG Cream 0 0 0 DEXA Increased basal energy expenditure (p ¼ 0.03) Insulin, glucose, leptin, CRP, LDL-, HDL-, total cholesterol, TAG, liver function and inflammation parameters (except sVCAM-1) n.i. Safflower oil Insulin, glucose, leptin, TAG, total and HDLcholesterol 41 25–31 3.2 50:50 c9,t11/ t10,c12 44 23–27 3.76 50:50 c9,t11/ t10,c12 43 25–35 3

2009 [109]

Body composition, insulin, glucose, bone mineral accretion, lipid profile, no diet or lifestyle restriction

7 months

2009 [107]

Body composition, markers of cardiovascular risk, no diet or lifestyle restriction

16 wks

2007 [104]

Body composition, markers of cardiovascular risk, no diet or lifestyle restriction

6 months

2007 [103]

Body composition, weight gain in holiday season, no diet or lifestyle restrictions

7 months

2007 [73] Body composition, markers of cardiovascular risk, no diet restriction, 3 x 45’ exercise/wk

14 wk

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2007 [106]

Body composition, markers of metabol syn, 500 mL skim milk CLA vehicle, no diet or lifestyle restriction

12 wk

50:50 c9,t11/ t10,c12

n.i.

500 mL skim milk

0

0

À Àc)

DEXA

Trunk fat mass reduction in overweight ( p ¼ 0.01), not in obese

Fasting glucose, TAG, LDL-, total cholesterol, blood pressure, insulin sensitivity, hepatic and renal function markers

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(Continued )

Table 1. (Continued)
Body composition þ/À effect on metabolic factors Increased LBM (þþ), decreased BF (þþ), and decreased insulin concentration in women only

Year ref. 62 22.5–24.2 3.9 50:50 c9,t11/ t10,c12 n.i. Sunflower oil 0 0 0 Skin fold thickness, DEXA, computerised tomography

Design and approach

Length of supplementation Isomers Form Placebo BM LBM BF Anthropometrics

Study finishersa)

Baseline BMI

CLA [g/d]

No effect on metabolic parameters vs. placebo Glucose, LDL-, HDL, total cholesterol, TAG, appetite, satiety, REE, substrate oxidation

2007 [37] Training subjects, body composition, blood lipid profile, REE, RQ, glucose tolerance, no diet or lifestyle restriction 48 30–35 3.2/6.4 50:50 c9,t11/ t10,c12 n.i. Safflower oil 0 ÀÀ DEXA # Decreased HDL (also in placebo), increased CRP and IL-6 (only in 6.4 g group) Decrease in waist circumference (þþ)

12 wk

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2007 [105]

Body composition, markers of cardiovascular risk, no diet or lifestyle restriction

12 wk

TAG, LDL- and total cholesterol, REE, RQ

ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
83 28–32 3.4 t10,c12 and c9,t11 n.i. olive oil 0 n.i. # DEXA, waist circumference Insulin, glucose, leptin, adiponectin, HbA1c, C-peptide 40 30–36 4.5 35–36% c9,t11/t10, c12; 1-2% c9,c11/ c10,c12; 1.5% t9, t11/t10,t11; <1% t8,c10/ c11,t13 50:50 c9,t11/ t10,c12 TAG Olive oil 0 " n.i. Olive oil À n.i. # Skin fold thickness, bioelectrial impedance, computed tomography Decreased limb (þþ; p ¼ 0.001) and torso-to-limb (þþ; p ¼ 0.017) skinfold thickness, decreased brachial artery endothelial function, increased F2isoprostanes # DEXA Insulin sensitivity, TAG, CRP, LDL-, HDL-, total cholesterol, leptin, adiponectin 83 >28 3.4 Lower BMI in cases Plasma glucose, insuvs. control at baseline lin, insulin resistance (þþ) 125 25–30 3.4 50:50 c9,t11/ t10,c12 TAG Olive oil ÀÀ 0 ÀÀ DEXA Reduced plasma cholesterol (total, LDL) and leptin, increased Lp(a), leuco- and thrombocytes 50:50 c9,t11/ t10,c12 TAG/FFA Olive oil À À/À À 3.4–3.6 þ/þþ À À/À À DEXA Decreased HDLcholesterol, hemoglobin (only within CLA-TAG group), leptin; higher Lp(a), thrombocytes and AST (only in CLA-FFA group) HDL-cholesterol, TAG, insulin, glucose, blood pressure, various blood parametres 157 25–30 Insulin, glucose, TAG, total cholesterol, ALT, blood pressure, heart rate

2007 [102]

Body composition, glucose and insulin changes, no diet or lifestyle restrictions

6 months

2006 [114]

Body composition, markers of cardiovascular risk, no diet or lifestyle restriction

12 wk

2006 [111]

2-center study, BM regain after 8wk restricted diet run-in, then modest hypocaloric diet (-1250 kJ dÀ1), no lifestyle restriction

12 months

2005 [97]

Body composition, open follow-up after 1y trial, no diet or lifestyle restriction

24 months

Long-term conjugated linoleic acid supplementation in humans

2004 [98]

2-center study, body composition, no diet or lifestyle restriction

12 months

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(Continued )

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Table 1. (Continued)
Body composition

Year ref. Isomers 50:50 c9,t11/ t10,c12 TAG High oleic sunflower oil 0 n.i. n.i. Air displacement Increased serum TAG ( p ¼ 0.053) and leucocytes Form Placebo BM LBM BF Anthropometrics 46 27–35 6

Design and approach

Length of supplementation Baseline BMI CLA [g/d]

Study finishersa)

þ/À effect on metabolic factors

No effect on metabolic parameters vs. placebo Glucose, insulin, insulin resistance, LDL-, HDL-, total cholesterol, various blood parameters

R. P Jutzeler van Wijlen .

2004 [115]

Body composition after 12w low-calorie diet, 16w calorie-controlled diet, which continued for 25w open-labelled, no lifestyle restrictions 82 25–30 1.5–3 c9,t11 or t10,c12 TAG High oleic sunflower oil # 0 DEXA #

12 months

ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Insulin, glucose, varius blood, liver and kidney function parameters 25 men 27–35 3 c9,t11 TAG Olive oil " # " Diameter measurement, bioelectrical impedance Impaired insulin sensitivity, higher 8-iso-prostaglandin F2a, 15-ketodihydroprostaglandin F2a ÀÀ Hydrodensitometry Glucose, insulin, Lp(a), TAG, FFA 54 27.8 1.8/3.6 22.6% t10,c12 23.6% c11,t13 17.6% c9,t11 16.6% t8,c10 7.7% t9,t11 7.7% t10,t12 11.9% others 50:50 c9,t11/ t10,c12 or t10,c12 FFA Olive oil #y/À Ày þ/þ TAG Oleic acid 0 þþ Increased REE (þþ) Glucose, insulin, TAG, FFA, energy and RQ (þ); intake increased satiety (þþ), fullness (þþ), reduced hunger (þþ) 57 men 28–33 3.4 À Ày/À Ày Bioelectrical impedance, sagittal abdominal diameter measurement Leptin, LDL-, total Impaired insulin sensitivity, increased cholesterol, TAG, FFA glucose (only in t10,c12 single-isomer group); decreased HDL-cholesterol 20 23 1.8 50:50 c9,t11/ t10,c12 n.i. Hydrogel # n.i. ÀÀ NIRIR light – –

2004 [179]

Body composition after 6 w run-in with 1.5 g sunflower oil, no diet or lifestyle restrictions

18 wk

2004 [32]

Body composition, insulin sensitivity, lipid peroxidation, no diet or lifestyle restrictions

12 wk

2003 [99, 180]

BM regain and composition after 3w verylow-calorie diet runin, no lifestyle restrictions

13 wk

2002 [180]

Lipid metabolism and body composition, effects on insulin sensitivity in men with metabolic syndrome (incl. insulin resistance), 4w run-in, no diet or lifestyle restrictions

12 wk

2001 [181]

Body composition, standard exercise program (3x week), no other lifestyle or diet restrictions 50 19–34

12 wk

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12 wk

4.2

n.i.

Olive oil

#

n.i.

ÀÀ

(Continued )

Eur. J. Lipid Sci. Technol. 2011, 113, 1077–1094
LDL-, HDL-, total cholesterol, Lp(a), various blood and liver function parameters, blood electrolytes, blood pressure Glucose, insulin, HDL-, total cholesterol, TAG, FFA, apolipoprotein A1 and Lp(a) TAG, total cholesterol, Lp(a), blood pressure, heart rate

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1083

subjects both male and female if not otherwise indicated. in subjects with BMI >30, no effect below BMI 30. c) in subjects with BMI <30, no effect above BMI 30. 0 ¼ no effect; þ/À ¼positive/negative trend ( p < 0.1); þþ/À À ¼significant positive/negative effect ( p < 0.05); #"¼tendency to decrease/increase compared to placebo with/without indication of level of significance. y within case group, but not between case group and placebo.
b)

No effect on metabolic parameters vs. placebo

tory genes was down-regulated and the expression of lipid metabolism genes up-regulated. These responses correlated better to obesity phenotype markers and fat percentage than BMI. Evidence is also growing that the effects of CLA on body composition may be region-specific. Raff et al. [107] measured less lower-body fat mass (9.5 kg vs. 10.2 kg, p ¼ 0.0008) and a concomitant higher lower-body LBM (14.7 kg vs. 14.3 kg, p ¼ 0.02) in women after 16 wk of mixed c9,t11 and t10,c12-CLA supplementation compared to placebo. Notably, such region-specific differences of body composition may be at least partly due to gender. The effects on BM and total fat mass were present, too, though somewhat smaller (70.2 vs. 71.1 kg, p ¼ 0.05 and 24.6 vs. 25.6 kg, p ¼ 0.02, respectively). Also in prepubertal children a greater attenuation in relative fat gains between the CLA treatment and placebo was observed in peripheral fat (À1.2 vs. 0.5%, p 0.001) and abdominal fat (À0.09 vs. 0.43%, p 0.02) [109]. Overall, the influence of CLA on body composition seems independent of physical activity. This point of view stands in opposite to the conclusion of a recent review [129]. But of the existing prolonged (!12 wk) studies only a minority did examine regularly training subjects [37, 73, 130]. Still, it is well-known that exercise has important implications on substrate oxidation and insulin sensitivity; and this may interact with the efficacy of CLA administration on body composition, glucose and lipid metabolism. It may therefore be an interesting approach for future studies to examine intervention designs combining CLA supplementation with exercise programs, known that also traditional energy-restricted weight-loss regimens combined with exercise generally work better than energy restriction alone. Seen that weight regain after energy restriction is also a common problem, it may be furthermore of interest to reinvestigate the observations of Kamphuis et al. [99], where after low-energy induced, initial BM loss, CLA supplementation induced a higher BM regain, but with significantly higher LBM. Undoubtedly, the most important difference between animal and human trials is dosage: most human studies used CLA in doses between 1.8 and 6.4 g/d. The median dose of 3.5 g/d used in the presented human trials (Table 1) reflects a mere 1.3% of energy of a 10 MJ/d diet, whereas the median CLA dose in pig studies was 27.3 g/d obviously constituting a far higher percentage of dietary energy [131]. When assessing the modulation of BM and body composition caused by CLA, it is deemed important to keep in mind that a placebo like, e.g. olive oil, containing non-conjugated diene-fatty acids, already may constitute an important change to the subject’s diet, modulate the lipid profile of its diet and consequently may have metabolic effects itself. To our knowledge, no author has corrected study results for such a possible bias or confounding effect hitherto. In support of this hypothesis are the recent findings from Norris et al. [108], which stated reduced trunk adipose mass and increased LBM, as well as lowered fasting glucose with safflower oil supplementwww.ejlst.com

Increased LDLcholesterol and apolipoprotein B, tendency to increased glucose

Body composition

Anthropometrics

Skin fold thickness, bioelectrial impedance

Bioelectrical impedance, DEXA

LBM

À Ày

BM

Form

Olive oil

Placebo

n.i.

Baseline BMI

27.5–39

Study finishersa)

Length of supplementation

55

2-center study, investigation on safety and body composition, no diet or lifestyle restrictions

Design and approach

Effects on anthropometric and metabolic variables, no diet or lifestyle restrictions

12 wk

Table 1. (Continued)

Year ref.

2000 [113]

2001 [31]

2000 [101]

Dose-response relationship on BFM, no diet or lifestyle restrictions

12 wk

47

28

1.7/3.4/ 5.1/6.8

CLA [g/d]

3.4

50:50 c9, t11/t10, c12

Isomers

50:50 c9,t11/ t10,c12

50:50 c9,t11/ t10,c12

n.i.

Olive oil

#

þþy

"

ÀÀ

BF

Ày

DEXA

Slight change of LDL-, HDL-cholesterol (clinically irrelevant)

þ/À effect on metabolic factors

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a)

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ation. Such an effect of safflower oil, when used as a placebo, can mask the true significance of any treatment effect such like CLA supplementation. The question merits further exploration what may be considered an appropriate fat source as a placebo. In summary, the outcome of the studies with CLA on body composition with a minimum duration of 12 wk (Table 1) indicates a reduction and a prevention of regain of BF in overweight to obese subjects. In this context, a metaanalysis restricted to prolonged (!12 wk) studies of CLA supplementation and its effect on body composition is warranted.

3.2 Safety aspects 3.2.1 Isomeric composition of CLA
Qualitative analysis of CLA often requires highly demanding analytical procedures and equipment. Relatively recent analytical developments allow detecting more positional and geometrical isomers of CLA [14, 132–135], and future studies on specific functional properties of different isomers are thus warranted. For example, eight geometrical and positional isomers of 9,11 and 10,12 CLA are possible, raising the question of how much they differ in manifestation and effect from each other. In addition, it is not a simplistic task to resolve the different isomers of CLA by GC combined with MS. Identified peaks may contain several isomers [136], misleading in terms of the expected isomer-specific activity and consequently constitute a confounder intriguing conclusions on isomer-specific and CLA efficacy in general. As reviewed by Gaullier et al. [137], the conjugation of linoleic acid may also result in the formation of cis,cis and especially trans,trans isomers that could accumulate progressively with time. Presently, clinical trials as well as industrial production concentrate on purified CLA preparations with a 50:50 mix of c9,t11 and t10,c12 in an average concentration of approximately 80%. In consequence, such a CLA mix entails low contents of other isomers with less than 5% of trans,trans and cis,cis forms. Nevertheless, how such a CLA ratio complies to a mixed and varied diet is yet unknown. Aside from safety, bioavailability is also an issue. A comparison of CLA in a 50:50 ratio of c9,t11 and t10,c12 isomers was performed, given in a milk-based drinkable form either as TAG, free fatty acids (FFA) or ethyl esters. Significantly less CLA was absorbed into chylomicrons over 6 h when given as ethyl esters, whereas the other two forms showed similar rates [138].

3.2.2 Glucose and insulin metabolism
Main safety concerns on CLA focus on glucose and insulin metabolism due to their TFA structural bond. Yet, whereas a diet extremely high in TFA (20%E) does indeed cause a significantly elevated postprandial insulin response compared
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to mono-unsaturated fatty acids (MUFA), no significant difference versus saturated fatty acids (SFA) in overweight ´ patients with type 2 diabetes is seen, as cited by Riserus et al. [139]. Nevertheless, the same author found in a more applicable design in regard to CLA supplementation (3 g/d c9,t11 CLA for 3 months, representing <1%E) a 15% reduction in insulin sensitivity in obese, non-diabetic men, but no differences in fasting blood glucose or serum insulin concentrations [140]. After ingestion of supplemental t10,c12, but not with the isomeric mix of c9,t11 and t10,c12 CLA, decreased insulin sensitivity was also reported in obese men with signs of the metabolic syndrome (BMI > 30), while fasting glucose and insulin did not again differ from control [23]. In two other reports from that same ´ study [21, 32] Riserus et al. again concluded insulin sensitivity to be impaired, together with enhanced oxidative stress by the supplementation of t10,c12 CLA. Yet, another study in humans with Type 2 diabetes showed a decrease in fasting plasma glucose after 8 wk of CLA supplementation with 6 g/ d (%50:50 ratio of c9,t11 and t10,c12) [25]. Fasting insulin, HbA1c, TAG, cholesterol and HDL were not significantly affected by CLA. No negative outcomes on fasting glucose and insulin after 16 wk of CLA supplementation with 6.4 g/d (%50:50 ratio of c9,t11 and t10,c12) were found in obese postmenopausal women with type 2 diabetes, either [108]. Notably, that study used safflower oil in a parallel design resulting in decreased fasting glucose. In consequence, a significant difference in fasting glucose and HOMA-R was observed between the CLA and safflower oil treatment. This underlines the importance of considering inherent glycemic effects of any dietary fat source used as a placebo in CLA supplementation studies. In contrast, another recent study with 3 g/d  8 wk of an identical isomeric mix of CLA found adverse effects on insulin and glucose metabolism in type 2 diabetic patients [35]. The majority of studies with increased CLA intake or supplementation have not found significant changes in fasting glucose, insulin and markers of insulin sensitivity in neither non-diabetic overweight to obese humans [96–99, 102–104, 113, 114, 141–144], nor in healthy, normal-weight humans [29, 31, 73, 79, 145, 146]. Rather in the opposite, improved plasma insulin concentrations [37] or increased insulin sensitivity [147] was demonstrated in healthy humans. Yet, the latter authors undertook also a similarly designed examination with an oral glucose tolerance test after 12 wk of mixed-isomer CLA supplementation in nine overweight, non-diabetic subjects. The authors stated a decreased insulin sensitivity, likely due to increased muscle ceramide content preventing insulin-stimulated activation of Akt/ Protein kinase B, thereby reducing glucose uptake in muscle [92]. But noteworthy, that study lacked a control group. In contrast, Lambert et al. [37] demonstrated an improved insulin response after an oral glucose load compared to a control group receiving high-oleic acid sunflower oil. Ahren et al. [148] found no effect on glucose, insulin and C-reactive
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peptide in healthy, young lean and obese as well as in old lean men, when supplementing either a CLA mix plus n3-PUFA or control oil. Only in older obese subjects a non-significant trend compared to control oil was measured towards increased basal rates of glucose, insulin and C-reactive peptide. Similarly, post hoc analysis showed subject-specific increased fasting serum insulin in the tertile with the highest waist circumference (94–109 cm) of postmenopausal women; no differences were found in the two lower tertiles [107]. Similar results were found for the HOMA-R calculations, suggesting that subjects with a large waist circumference are more sensitive to potentially negative effects of CLA supplementation. Seemingly, both age and adiposity may modulate the effects of CLA as well as of n3-PUFA. These outcomes again scrutinize the choice of placebo and demand to control for it. A long-term study examining the safety of 6 g/ d  12 months of the CLA mix in obese subjects found no effects on insulin or liver function over the long term [149]. Hitherto, three studies were undertaken over a 1 year period [97, 98, 111], one of which was extended as a followup in an open-labelled design to a total supplementation length of 24 months. All of them add with their results to the safety of CLA regarding insulin sensitivity and lipoprotein metabolism. Still, in some cases single-isomer administration did show a potential adverse influence on insulin sensitivity, whereas a CLA mix seemingly does not influence negatively glucose and/or insulin metabolism in healthy subjects. The effects on diabetic subjects remain controversial, but data should be looked at with caution since differences in measuring methods may have played an important role in the controversial outcomes. Newer data from a 6-months study using euglycemic hyperinsulinemic clamp method [102], looked upon as the gold standard method, add to the safety of CLA. In spite of the evidence rather indicating no negative effects – if not positive – also on the sub-group of type 2 diabetics, insulin sensitivity implies a delicate conflict. Typically this subgroup is also confronted with overweight or obesity the most, hence belonging to the main target group for potential anti-adipogenic effects of CLA. Further research to exclude any safety risks for diabetic subjects with or without metabolic syndrome is therefore essential.

3.2.3 Liver metabolism and lipid profile
In contrast to animal studies, no adverse effects on liver safety parameters from CLA supplementation are reported in humans [115] up to date. Except in 2009, when a single case report revealed a possible incidence of toxic hepatitis after supplementation with CLA [150]. Unfortunately, the report did not mention the daily dosage and the isomeric composition of the CLA supplement used by the patient. In other studies with healthy humans no clinically relevant adverse effects, albeit no health-promoting either, have been
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noted on serum lipid profile [29, 104, 110, 121, 144, 146, 151]. Another safety concern is that CLA may reduce breast milk fat, and its use therefore is discouraged in breast-feeding women [152]. However, newer data from the same group [153, 154] did not support those results. A study in healthy humans found a reduction in plasma TAG, low (LDL) and very-low density lipoprotein (VLDL) cholesterol, and VLDL–TAG concentrations after 8 wk of supplementation with two different isomeric mixes of CLA [155]. Although significance was not reached in any criteria, these effects have to be looked upon as health-promoting, if relevant at all. On the other hand, 4.6 g/d of a CLA mix supplement for 16 wk in overweight, postmenopausal women increased TAG (þ15%, p < 0.01), lowered HDL (À14%, p < 0.01) and the ratio of total cholesterol:HDL (þ11%, p < 0.05) compared to a c9,t11 CLA-enriched milk (5.1 g/d) and olive oil (5.5 g/d) [156]. However, it has to be noted that such high amounts of dietary CLA are unrealistic to achieve even by a strictly focused diet. The same is true for a recent study comparing iso-energetic diets with oleic acid, industrial TFA and a mixture of c9,t11 and t10,c12 CLA at dosages of 66.2 g/d, 21.8 g/d and 26.8 g/d, respectively, which is not achievable by a diet [157]. However, the lipoprotein levels in subjects (n ¼ 61, BMI ¼ 22.8 þ 3.2) worsenend after both the industrial TFA and CLA diet in relation to the oleic acid diet. The question remains, if the added statistical power by using high dosages is not outbalanced by the short study duration and the lack of a washout period (three consecutive periods of 21 days). Furthermore, the same author concluded previously that the daily intake of 19.3 g of a c9,t11/t10,c12 CLA mix (ratio 80:20) for 3 wk does not produce clinically relevant effects on markers of liver and kidney function in healthy humans [158]. Belury et al. [159] demonstrated a significantly inverse correlation of plasma CLA with BM and serum leptin in humans with type 2 diabetes, allied with a stronger linkage of t10,c12 than of c9,t11 CLA. Medina et al. [79] measured a transient decrease in circulating leptin in healthy women during 7 wk of CLA supplementation. However, there was no change in appetite parameters around the time of the greatest decrease in leptin, and levels returned to baseline during the final weeks of that study. No changes in serum leptin, total cholesterol, HDL, LDL, FFA and TAG were reported after 6 wk of CLA supplementation in women [121]. Similarly, supplementation of either 3 g/d of c9,t11 or t10,c12 CLA did not affect the serum lipoprotein profile in slightly overweight humans with LDL phenotype B [146]. Moloney et al. [35] found an improvement in HDL and LDL cholesterol concentrations after CLA supplementation with 3 g/d of the isomeric mix, resulting in a possible clinical benefit due to a reduction of the LDL:HDL ratio of 14.5%. No TAG reduction could be found in that study, but a negative glucose and insulin response. Taylor et al. [114] did not find any significant changes in lipid profile either, but an impaired brachial artery endothelial function
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consistent with an increased cardiovascular risk, in 40 healthy overweight individuals. Furthermore, the consumption of dairy products naturally high in CLA and VA did neither significantly change blood lipid profile nor cardiovascular disease risk factors in healthy, middle-aged men [29]. The tailored production methods and the intake of 500 mL full-fat milk, 12.5 g butter, and 36.3 g cheese per day resulted in an additional intake of 1.5 g CLA and 4.7 g VA, along with additional 150 kcal compared to control (2562 kcal/d) or to a ‘normal’ diet, respectively. The inclusion of human bioconversion of VA as suggested elsewhere [17] may provide an additional %940 mg of CLA and sum up into a probable total of %2.4 g available CLA in those subjects. Similar results, i.e. no changes in risk factors of the metabolic syndrome, blood lipids, renal and hepatic function markers were reported in overweight and obese individuals with metabolic syndrome after 12 wks’ intake of 500 mL skim milk with 3 g/d of CLA [106]. In a quantitative review the regression lines for the changes both in LDL, HDL and the ratio of LDL:HDL cholesterol were steeper from industrial TFA intake than for ruminant TFA or CLA, though far from being significant ( p ¼ 0.42 from ruminant vs. industrial, p ¼ 0.92 for CLA vs. industrial) [160]. Therefore, the authors speculate that all TFA with one or more bonds in the trans configuration raise the ratio of LDL to HDL cholesterol irrespective of their origin or structure. However, it was already previously concluded that no differences in the risk of coronary heart disease are evident between total, ruminant, and industrial TFA when the intake is below 2.5 g/d [161]. Furthermore, it is stated elsewhere that it is not possible to identify differences between TFA from ruminant and industrial fat in their effects on metabolic risk parameters, due to the lack and the impractibility of human intervention studies [162]. One reason is simply because the amount of TFA from dairy sources used in risk assessment studies greatly exceeded the intake of ruminant fat in usual diets. Thus, TFA from natural sources consumed in actual amounts seem not to contribute to the risk of coronary heart diseases [163]. This opinion was recently reinforced in a casecontrol study where no increased risk of a first acute myocardial infarct could be observed with higher intake of dairy fat [164]. Remarkably, in spite of a 15% higher dairy fat intake of the control group (26.3 vs. 22.9 g/d), total SFA intake was similar (35.1 vs. 35.8 g/d), but the ratio SFA: dairy fat (1.33 vs. 1.56) and total TFA intake (3.8 vs. 4.7 g/d) were lower, i.e. advantageous in the higher-dairy fat diet. Similarly, a recent study stated the most favourable lipid profile in subjects with an intake of 1.5%E (4.2 g/d of a 2500 kcal diet) as ruminant TFA compared to 3.7%E from ruminant or industrial TFA or even a control diet with low-TFA (0.8%E from any source) [165]. It has to be noted that those iso-energetic diets provided a rather high 37%E from fat in regards to present nutritional recommendations. Nevertheless, 4.2 g/d of ruminant TFA is an achievable
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amount through a high intake of dairy and meat products. Comparing the plasma fatty acid profiles in meat-eaters, vegetarians and vegans in a British study demonstrated that eating meat caused a much higher plasma content of RA compared to vegetarians (67%) and vegans (38%), and of t10,c12 CLA (94 and 81%, respectively) [166]. Such outcomes debilitate the often heard provisos against full-fat dairy food consumption, i.e. concomitant high intake of SFA and TFA. Even more, a recent study indicates that CLA may reduce total SFA as well as total n-6 fatty acids and especially AA [167]. In conclusion, all of these results provide further support for the theory that dietary ruminant TFA at a high, but naturally achievable intake has no detrimental effect on blood lipids.

3.2.4 Oxidative, inflammatory, and immunomodulating potential
Because CLA is easily oxidised, there have been considerable safety concerns about its supplementation. Basu et al. showed twice [22, 168] that supplementation with mixed c9,t11 and t10,c12 CLA of 4.2 g/d in middle-aged men with abdominal obesity during 1 month, but also in healthy humans for 3 months, may induce both non-enzymic and enzymic lipid peroxidation; yet, without any side effects. Hence, a significant increase in serum g-tocopherol in the 3 months’ study [22] was observed. Further studies were suggested on the mechanisms and consequences of the increase in lipid peroxidation after CLA supplementation. The intake of purified t10,c12 CLA was found to increase C-reactive protein (CRP), a marker for inflammation, whereas the CRP increase after supplementation of an isomeric mixture of c9,t11 and t10,c12 CLA was not significant [21], or within the experimental group only [103]. Moreover, this latter study also showed decreased levels of soluble vascular adhesion molecule (sVCAM-1) compared to control, actually rather referring to a reduced endothelial inflammatory status. On the other hand, urinary F2-isoprostane, another marker of oxidative stress, was increased compared to placebo for both t10,c12 CLA and the isomeric mix in the former study [21]. Similarly, no effect on CRP but increased urinary F2-Isoprostane was measured in overweight, young, and healthy men when receiving a diet enriched with 4.6 g/d of the isomeric CLA mix [144]. Except of this sign of increased free radical-induced lipid peroxidation the authors did not detect changes in other inflammatory, atherosclerotic or diabetic risk markers. An increase in CRP after supplementation of the isomeric CLA mix was found again in three other studies [104, 156, 169], but no effects on other inflammatory markers such as tumour necrosis factor (TNF-a) were measured. An increased CRP concentration, even at sub-clinical levels, is associated with the presence of the metabolic syndrome and predicts incident myocardial infarction, stroke, peripheral arterial disease and sudden cardiac death. Moreover, CRP and TNF-a
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have not only been associated with the metabolic syndrome, but also with several important steps for its development. In contrast, a study with three different doses in a consecutive manner of both the single isomers c9,t11 and t10,c12 did not find an effect on CRP at all [55]. A dose-dependant effect can only partly explain these contrary outcome, because the lacking effect of the highest dose of t10,c12 CLA in that study (2.52 g/d) stands in contrast with elevated CRP not only at a higher dose (3.4 g/d) of the same purified isomer [21], but also at a lower dose of an equal mix of both isomers (2.1 g/d ffi 1.05 g/d t10,c12) [169]. The other part of the explanation may be a lower BMI (24.6 þ/À 0.4) in the study of Tricon et al. [55] compared to the two others, seen that CRP levels are also determined by BF mass [170]. In this context, it is also reminded that adiposity in general is associated with an increased inflammatory status [171]. In a review on CLA and its effects on human health, Whigham et al. [172] concluded that CLA may have implications on promoting maintenance of LBM during immune stimulation, especially in wasting diseases such as cancer or AIDS, and during late stages of autoimmune diseases such as Lupus. CLA may also help downregulate type 1 hypersensitivity reactions, leading to less severe immune responses to allergens. Both c9,t11 and t10,c12 CLA were found to reduce mitogen-induced lymphocyte activation in a dose-dependant manner, but did not affect lymphocyte subpopulations, cytokine production or CRP [55]. The authors presumed that a reduction in lymphocyte activation may be undesirable in terms of host defence, whereas it may be useful in situations such as allergy and inflammatory diseases. Beneficial effects on immune function after CLA supplementation were demonstrated in a study in young, healthy volunteers finding increased levels of IgA, IgM and anti-inflammatory IL-10 along with a concomitant decrease in IgE, pro-inflammatory cytokines, TNF-a and IL-1b [173]. In summary, data for the expected anti-inflammatory effects and a supportive role of CLA on immune function is inconclusive. It is in particular unclear how a changed immune status or inflammatory disease factors, like e.g. obesity, influence the outcome of CLA supplementation in this respect. There is also supporting evidence with regard to the maintenance of bone mass by CLA, probably partly linked with its anti-inflammatory potential (see review: [44]).

3.3 Implications regarding CLA supply and intake
As demonstrated elsewhere, reports on human CLA intake vary, and usually do not consider food production methods and VA-induced CLA biosynthesis in humans [17]. Food products from natural grass-fed ruminants have a several-fold higher CLA and VA content and, along with consideration of endogenously formed CLA from VA, result in a substantially higher CLA availability than previously estimated. Exogenous CLA supply from milk, cheese, lamb, and beef from grass-based ruminant production methods was conserß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

vatively calculated to be about double that of estimates based on modern production methods. Using available human consumption data resulted in an estimated achievable CLA intake (including VA bioconversion) of between 711 and 1107 mg/d [17]. The conservative nature of this result was confirmed recently, when 1170 mg/d of total dietary CLA from naturally enriched food (derived from grass-based feeding strategies) was measured [174], even without taking into account VA biosynthesis in humans. Furthermore, a lower CLA intake than naturally achievable may be a rather general phenomenon nowadays where energy-restricted dieting is normal, and dietary fat balance possibly is impaired by reduced animal/dairy fat because of shifting culinary habits with the concomitant high intake of industrial plant oils from snack foods. From a nutritional point of view it may be interesting to determine if CLA may be looked at as a marginal missing nutrient in present food sources, rather than a dietary supplement. This raises the question if an enrichment of foods with CLA in similar doses and isomeric profile like naturally found – considering animal-adapted feeding regimens – can account for a correction of the presently low content caused by non-adequate feeding of animals and contemporary changes in our nutrition and lifestyle. How these data add to a thinking-over of the actual nutritional fat – especially of animal origin – and meat intake recommendations has still to be discussed, but it certainly cannot be neglected, and must not be equated with the predominating meat and fatty acid profile of nowadays’ nutrition. To increase CLA intake in the human diet may not only be potentially advantageous because of its effect on body composition as reviewed, but also in respect to the health aspects linked with the consumption of its main food carrier, the dairy products. High intake of dairy foods and consequently high milk calcium is supposed to be correlated with lower prevalence of obesity, bone density, and – potentially related with higher CLA content in full-fat dairy foods – decreased risk for colorectal cancer [175]. Simply giving pasture access to ruminants as much as possible would be a meaningful and easy approach, enhancing not only CLA and VA content of meat and dairy products, but furthermore such a practice would produce a fatty acid profile closer to nutritional recommendations regarding MUFA, PUFA, SFA and n-3 fatty acids [176, 177]. Considering a more selective, all-pasture derived food choice providing about 35–40%E from fat, even >3 g/d of total available CLA seem realistic [178]. For the consumer an adequate daily consumption of full-fat dairy foods from carefully chosen sources would be mandatory.

4 Conclusion
Reviewing the outcome of CLA supplementation on body composition from studies lasting !12 wk revealed a considerable variation in design and results. Nonetheless, the results of studies on body composition indicate a modest reduction
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and/or prevention of regain of BF in overweight/obese subjects. In an environment where steady fat accumulation is the norm and adiposity is a pandemic problem, even modest improvements on BF mass as potentially achievable with CLA could be relevant. Impaired insulin sensitivity induced by supplemental CLA remains a safety concern, yet results are inconclusive, and seemingly restricted to diabetics and/or single-isomer application. A meta-analysis of only extended studies is warranted; therewith paying attention to a slower metabolic rate of humans compared to animals and the different premises of depot fat reduction or reduced de novo fat deposition. Future intervention studies should also take into account genderrelated differences in lipid metabolism, genotype effects and apply adequate run-in control periods in order to identify possible inherited effects of placebo like, e.g. olive or safflower oil. Such effects may have intrigued a part of the results in the reviewed studies on body composition as well as on safety. Future research will have to elucidate if CLA, and in which isomeric composition, may be considered as a marginal missing, semi-essential nutrient in our present diet. We thank Alexandra Schmid, Agroscope Liebefeld-Posieux, Swiss Federal Research Station for Animal Production and Dairy Products (ALP), Schwarzenburgstr. 161, 3003 Berne, Switzerland, for her kind and appreciated editorial assistance. This research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors. The author R. Jutzeler van Wijlen is head of R&D at Sponser AG, sports food manufacturer.

[8]

[9]

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