Cla PDF
Cla PDF
Cla PDF
Bethesda, Maryland
Sponsors
Mitsubishi Corp.
Natural ASA
BASF
PERSPECTIVES ON
Contents
1 AGENDA
5 SPEAKER ABSTRACTS
OVERVIEW OF CLA:
B I O C H E M I S T R Y A N D M E TA B O L I S M
B I O L O G Y / H E A LT H E F F E C T S
B. Cancer Effects
C. Other Areas
HUMAN TRIALS/EFFICACY
27 CHAIRPERSONS
31 SPEAKER LIST
37 AT T E N D E E L I S T
PERSPECTIVES ON
Agenda
OVERVIEW OF CLA:
B I O C H E M I S T R Y A N D M E TA B O L I S M
Pamela Starke-Reed, DNRC/NIH – Chair
10:35 am Break
1
Agenda
B I O L O G Y / H E A LT H E F F E C T S
12:05 pm Lunch
B. Cancer Effects
John Milner, NIH/NCI – Chair
3:45 pm Break
2
Agenda
C. Other Areas
Deborah Applebaum-Bowden, NIH/NHLBI – Chair
9:40 am Break
3
Agenda
HUMAN TRIALS/EFFICACY
Beth Yetley, FDA – Chair
3:15 pm Break
4
Speaker Abstracts
The Origin of CLA Dale E. Bauman
Cornell University
Department of Animal Science
The CLA in foods derived from ruminants relates to synthesis involving ∆9-desaturase and trans-7 C18:1
the biohydrogenation of unsaturated fatty acids by produced in the rumen. Other CLA isomers in milk fat,
rumen bacteria and most of the work has involved which are present in much lower quantities, originate
dairy cows and milk fat. cis-9, trans-11 CLA is the from rumen biohydrogenation. Under certain dietary
predominant isomer representing 75 to 80% of total conditions, a portion of linoleic acid biohydrogenation
CLA. This isomer is formed as an intermediate in the in the rumen can involve an isomerization of the
biohydrogenation of linoleic acid. Although rumen cis-9 double bond to form trans-10, cis-12 CLA.
production is the source for a portion of milk fat CLA, These diets are associated with a change in the
the major source is endogenous synthesis. Between 70 rumen environment, an increase in milk fat content
to 95% of the cis-9, trans-11 CLA in milk fat originates of trans-10, cis-12 CLA, and a marked reduction in milk
by endogenous synthesis via ∆9-desaturase from fat secretion. Overall, milk fat content of CLA is largely
trans-11 C18:1, another biohydrogenation intermediate. dependent on rumen outflow of trans-11 C18:1 and
In ruminants, ∆9-desaturase activity is high in adipose tissue activity of ∆9-desaturase; both of these variables
tissue of growing animals, and in mammary tissue and can be markedly affected by diet and vary substantially
adipose tissue of lactating animals; mRNA and protein among individuals. Thus, by manipulating the diet and
for this enzyme are negligible in liver. The second most through genetic selection, the CLA content of foods
prevalent CLA isomer in milk fat is trans-7, cis-9 and derived from ruminants can be altered.
it originates almost exclusively from endogenous
PERSPECTIVES ON
6
Conjugated Linoleic Acid Research
Documentation of CLA Intake in Humans; Michelle Kay McGuire and Mark A. McGuire
What We Know and What We Should Know
Washington State University and University of Idaho
Because of the clear potential for various isomers of Because of our interests in infant and child health,
CLA to influence human health, documentation of we have also documented CLA intakes in these groups.
dietary CLA in the human diet is of interest. Various We and others have documented that human milk
methodologies have been utilized to quantify intake of contains a variety of CLA isomers in relatively high
CLA, including the use of disappearance data, dietary concentrations, potentially resulting in quite high
recalls, food frequency questionnaires, weighed food CLA intake by breastfed, but not formula-fed babies.
records and biochemical analysis of food duplicates. More recently, we studied school-aged children (5-15
These methodologies all have limitations, although yr; n = 40) and documented total CLA and c9,t11-CLA
the analysis of food duplicates is considered the gold intakes by weighed 3-d records. Data suggest relatively
standard at this time. For example, accuracy of all of high intakes of CLA in this age group. No relationship
the indirect methods relies heavily on the accuracy of between age and absolute CLA intake was found;
a database containing the CLA contents of commonly relative to body weight, CLA intakes were highest in
consumed foods. Although a substantial amount of the youngest children. Interestingly, girls consumed
work was conducted initially to document CLA in significantly more CLA than did boys (184 and 158
various foods, our database remains limited. Further, mg/d, respectively). The physiologic consequences of
although a growing literature suggests that the various CLA intake throughout the lifespan are currently not
CLA isomers influence human health differently, very understood. However, early programming during fetal
little data are published concerning the isomeric CLA growth, infancy and childhood might decrease risk for
contents of foods. None-the-less, researchers utilizing chronic disease in later life. Thus, a better and more
indirect methodologies have estimated CLA intakes in accurate understanding of CLA intakes and factors
various locations including the United States, Australia, influencing CLA consumption throughout the lifespan
German and Finland; typical intakes are reported to might lend insight into what might be considered
range from 50 to 1000 mg/d. Using food duplicate appropriate dietary recommendations for this potential
methodology, we have also documented that “total nutrient. Further, this information is needed to better
CLA” intakes are 212 and 151 mg/d in adult men and delineate which effects of CLA might be realized from
women, respectively; c9,t11-CLA intakes were found to dietary intake, and which effects can only be obtained
be 193 and 140 mg/d in men and women, respectively. from supplementation.
Estimates by food duplicate methodology are consis-
tently lower than those collected with food records.
PERSPECTIVES ON
7
Conjugated Linoleic Acid Research
Metabolism of Conjugated Linoleic Acid Sebastiano Banni
Among 28 possible conjugated linoleic acid (CLA) In adipose and mammary tissues the metabolites
positional and geometrical isomers only the 9cis, content ranges from 5 to 15% of total CLA, and in
11trans and the 10trans, 12cis have been extensively plasma and liver from 10 to 30%. Other metabolites
tested for biological activities. with 16 carbon atoms, conjugated 16:2 and 16:3,
deriving most probably from peroxisomal beta oxidation
Both these CLA isomers have been shown to undergo of CLA and its metabolites respectively, have been
elongation and desaturation processes similar to those detected. This suggests an efficient metabolism of
occurring with linoleic acid, in a variety of animal CLA and its metabolites in peroxisomes
species and also in humans, retaining the conjugated
diene structure. Thus, CLA seems to interfere with As a polyunsaturated fatty acid that gives rise to
linoleic acid metabolism, and thereby with arachidonic 20 carbon atoms metabolites, CLA metabolism may
acid deposition, particularly in those tissues where interfere with eicosanoid formation by different ways,
CLA and some of its metabolites, conjugated 18:3 1) by decreasing arachidonic acid supply, 2) by
and conjugated 20:3 acid are preferentially incor- interfering with lipoxygenase and cyclooxygenase
porated such as adipose and mammary tissues because pathways, 3) by forming eicosanoid-like molecules
of their higher incorporation into neutral lipid. On which may then compete with regular eicosanoids.
the other hand, conjugated 20:4 is preferentially
incorporated into specific phospholipids mainly
phosphatidylinositol and phosphatidylserine.
PERSPECTIVES ON
8
Conjugated Linoleic Acid Research
Concepts for Development of an Analytical Method Martin P. Yurawecz, Kim M. Morehouse
to Determine CLA Composition in Foods, Dietary and Pierluigi Delmonte
Supplements and Reference Materials
Center for Food Safety and Applied Nutrition
U.S. Food and Drug Administration
Current qualitative and quantitative determination maxima for each type of c/t isomer are different.
of conjugated linoleic acid (CLA) isomers in foods, A procedure will be described that greatly improves
dietary supplements and reference materials involves both the identification and quantitation of CLA
the complementary use of both GC, with FID or isomers based on their HPLC retention volumes
MS detection, and silver ion (Ag+) HPLC with UV relative to toluene, and the use of secondary
detection. To date, the identification of CLA isomers internal standards containing well characterized
has been performed by using Ag+ HPLC with GC UV chromophores (max. and coefficients).
confirmation, or visa versa. An internal standard This will simplify the quantitation using Ag+
was used to quantitate the total CLA by GC, and Ag+ HPLC, which is the technique that provides the
HPLC quantitation was calculated from the GC data best separations.
based on the type of isomer either c,c or t,t or c/t.
This type of analysis, utilizing the GC data to
quantitate the Ag+ HPLC data, was necessary because
both the extinction coefficients and the absorbance
PERSPECTIVES ON
9
Conjugated Linoleic Acid Research
Obesity and Lipid Metabolism: Body Fat James P. DeLany
Conjugated linoleic acid (CLA) has been shown products. The Wisconsin group first showed, and we
to reduce body fat accumulation in several animal confirmed that the active isomer responsible for the
models. We have conducted several studies in AKR/J reduced body fat accumulation is the t10,c12 isomer.
mice showing that CLA reduces body fat accumulation The potential negative effects of CLA, namely increased
whether animals are fed a high fat or low fat diet, with liver weights and increased insulin levels were also in
no effect on food intake. One mechanism by which response to the t10,c12 isomer. We have shown that
CLA reduces body fat is by increased energy expendi- a dose of t10,c12 as low as 0.15% is effective in
ture, which is observed within 1 week of CLA feeding reducing body fat while animals were on either a low
and is sustained for at least 6 weeks. The increased fat or a high fat diet. We have also demonstrated that
energy expenditure is sufficient to account for the CLA is effective in reducing body fat in older mice,
decreased fat accumulation. We have observed who were already obese. All of the previous work
increased fat oxidation but no decrease in de novo had been done in young, growing animals, and these
fat biosynthesis with CLA feeding. All of the early CLA studies showed that CLA is effective in older animals
studies were undertaken using a synthetic preparation as well, which would have implications in the use of
containing approximately equal amounts of the major CLA in humans. The published human studies with
isomer found in beef and dairy (c9t11) as well as CLA have shown mixed results.
another isomer (t10c12) which is very low in natural
PERSPECTIVES ON
10
Conjugated Linoleic Acid Research
CLA Effects on Adipocytes: Michael W. Pariza
Mechanistic Considerations
Food Research Institute
Department of Food Microbiology and Toxicology
University of Wisconsin-Madison
Conjugated linoleic acid (CLA) exhibits a number of to produce the multitude of biological/physiological
seemingly disparate biological/physiological effects effects that are attributed to CLA. CLA-induced
including inhibiting carcinogenesis at several stages reduction in body fat gain is an example of a
in experimental animals, reducing atherosclerosis, single-isomer effect that is due specifically to the
reducing body fat gain, and enhancing immune trans-10, cis-12 isomer. There are two aspects to
function while reducing the catabolic effects of elucidating the biochemical mechanism(s) that
immune stimulation. The biochemical mechanisms underlie this observation: determining if trans-10,
that underlie these observations are emerging from cis-12 CLA acts directly or via a metabolite to
research in a number of laboratories. These mechanisms regulate lipid accumulation in adipocytes; and
originate with the isomers of CLA, in particular the identifying the signaling pathways through which
cis-9, trans-11 and trans-10, cis-12 CLA isomers, trans-10, cis-12 CLA (or its bioactive metabolite) act
both of which have been shown to exhibit biological to control body fat gain in vivo. New findings that
activity. Emerging evidence indicates that these address these issues will be presented.
CLA isomers act both independently and in concert
PERSPECTIVES ON
11
Conjugated Linoleic Acid Research
Conjugated Linoleic Acid Isomers Lance H. Baumgard 1 and Dale E. Bauman2
and Mammary Lipid Metabolism
1 University of Arizona, Tucson, Arizona
2 Cornell University, Ithaca, New York
Abstract Text: Supplemental conjugated linoleic acids (ACC, FAS & ∆9-desaturase) required for de novo fatty
(CLA) reduce milk fat synthesis in lactating cows, sows acid synthesis. Furthermore, mammary lipogenic
and women. CLA effects are specific for fat as other capacity, as measured by labeled acetate incorporation
milk components are unchanged. We have demonstrated into lipid, was dramatically reduced (>80%) when cows
effects on mammary lipid metabolism are the result of received exogenous trans-10, cis-12 CLA. At low doses
trans-10, cis-12 CLA, as similar amounts of exogenous trans-10, cis-12 CLA equally reduces the yield of de
cis-9, trans-11 CLA have no effect on milk fat parame- novo and preformed fatty acids. Consistent with this
ters. Abomasal infusion of purified trans-10, cis-12 at CLA decreases the expression of enzymes responsible
a rate of 3.5 to 14.0 g/d decreases milk fat yield by for uptake and intracellular transport of preformed
25 to 50%, respectively. The mammary gland is more fatty acids (LPL & FABP), which largely explains how
sensitive to CLA than adipocytes as the amount of CLA CLA decreases the milk fat content of lactating sows
required (0.016% of diet) to substantially reduce milk and nursing women, two species where utilization of
fat synthesis is much lower than needed (0.5 to 1.5% preformed circulating lipids is the predominant source
of diet) to reduce the body fat content in growing of milk fatty acids. In addition, trans-10, cis-12 CLA
animals. Examination of the milk fat composition reduces the mRNA expression of enzymes involved in
demonstrates CLA causes a reduction in secretion of fatty acid esterification (GPAT & AGPAT). However, the
all fatty acids, but those of de novo origin are more amount of CLA required to reduce milk fat synthesis
extensively affected. On a molar basis, ~80% of the in lactating cows has little or no effect on circulating
decrease in milk fat yield can be explained by a metabolites (NEFA, glucose & ß-hydroxybutyrate) or
reduction in fatty acids synthesized within the hormones (insulin & leptin) associated with bioener-
mammary gland. In addition, using substrate/product getics. It is thought that other specific CLA isomers
ratios as a proxy for the ∆9-desaturase it is evident (i.e. trans-8, cis-10 CLA) or conjugated trienes may
that CLA inhibits this enzyme. Consistent with changes alter milk fat synthesis but they have not yet been
in milk fatty acid composition, we demonstrated trans- tested in pure form.
10, cis-12 CLA reduces mRNA expression of enzymes
PERSPECTIVES ON
12
Conjugated Linoleic Acid Research
PPARS as Potential Mediators Jack Vanden Heuvel
Conjugated linoleic acids (CLA) are a group of position- three members of this receptor family are activated by
al and geometric isomers of linoleic acid (LA). Interest CLA isomers, although their affinity for PPARα is much
in these dietary fatty acids stems from the fact, unlike greater than for β and γ. The ability to activate PPARα
LA, CLA is protective against cancer, atherosclerosis may help explain CLA’s effect on hepatic fatty acid
and diabetes in a variety of animal models and in metabolism. Since the biological role of PPARβ is not
some preliminary human trials. Despite the plethora of well established, it is difficult to determine if activa-
studies showing the beneficial properties of CLA, there tion of this receptor may explain the health benefits
is a paucity of mechanistic information on how this of CLA. We have focused our recent attention on PPARγ
compound exerts its effects. Also, there has been because of its beneficial role in diabetes, inflammation
little detailed exploration of how the various isomers and cancer. Although CLA isomers are weak ligands for
differ in their biological effects. The tissue and isomer PPARγ, we have shown that this receptor is essential
specific effects raise the possibility that CLA requires for these fatty acids to regulate gene expression in
interaction with a cognate receptor to produce its the macrophage and in the adipocyte. The possibility
response. We hypothesized that CLA causes its positive exists that CLA isomers require metabolism to become
effects by regulating gene expression subsequent to an active PPARγ ligand. Together, these studies have
binding to one (or perhaps several) fatty acid-regulated identified activation of PPARγ as a possible mechanism
transcription factor(s). In particular, we have focused by which CLA can regulate gene expression and
on nuclear receptors (NRs) implicated in fatty acid ultimately result in its beneficial effects. This detailed
regulation of gene expression, the PPAR family molecular information on how CLA results in its health
(α, β/δ and γ). Each PPAR subtype has evolved to benefits in animal models may assist in determining
fulfill a different biological niche and are targets of the benefit of supplementation of CLA in humans.
important hypolipidemic and anti-diabetic drugs. All
PERSPECTIVES ON
13
Conjugated Linoleic Acid Research
Toxicology Studies on Clarinol Sue O’Hagan
Conjugated linoleic acid (CLA) is found naturally in 90-days. The material was tested at a dose level of
foods such as dairy and meat products. In nature the 1%, 5% and 15% in a synthetic diet (AIN-93G). A high
c9, t11 isomer predominates. Commercial preparations fat control diet containing 15% safflower oil was also
contain a mixture of isomers, with c9, t11 and t10, tested in the study. In keeping with the findings from
c12 often occurring in equal proportions. In addition, other studies on CLA, Clarinol was found to cause liver
the potential intake from commercial sources of CLA enlargement. The pathology data indicate that this is
is higher than that from the diet. A program of an adaptive effect that occurs only in female rats with
toxicology studies was therefore conducted to confirm high doses of CLA, and is reversible upon withdrawal
the safety of a preparation containing a mixture of of the test material. A No Observed Adverse Effect
CLA isomers. Level was identified in the study.
PERSPECTIVES ON
14
Conjugated Linoleic Acid Research
Safety Assessment of Conjugated Schulte, S.; Pfeiffer, A.M.; Rensmann F.W.;
Linoleic Acid (CLA) Esters for the Braun, J.; Hasselwander, O.; Kaesler, B.
Use as Feed Additive in Pigs
BASF AG Ludwigshafen, Germany
CLA is a generic term describing different naturally Results from the available studies, which will be
occurring isomers of linoleic acid with 2 conjugated presented, indicate that CLA esters used as feed
double bonds. The two primary CLA isomers have c9,t11 additive at concentrations up to 0.5% in animal
and t10,c12 configuration. Beneficial effects of CLA feed are safe.
such as change in body composition, chemoprevention
and improved insulin sensitivity have been reported in
animals.
PERSPECTIVES ON
15
Conjugated Linoleic Acid Research
CLA and Mammary Cancer Prevention Research Clement Ip, Ph.D.
There is strong evidence that CLA is an effective work showing that CLA treatment leads to the modula-
anticancer agent in the animal model. Although a tion of a panel of biomarkers which are suggestive of a
number of cancer sites have been shown to be protect- decrease in proliferation and an increase in apoptosis.
ed by CLA, tumor development in the mammary gland Dairy products that are enriched in 9,11-CLA are of
appears to be particularly sensitive to CLA intervention. special interest to the food industry. Vaccenic acid,
This may be due in part to the preferential accumula- an intermediate in the biohydrogenation of linoleic
tion of CLA in neutral lipid of adipocytes, which acid in the rumen, is also high in cow’s milk. There
represent the predominant cell type in the mammary is emerging data that mammals have the ability to
tissue. CLA stored in adipocytes could conceivably convert vaccenic acid to 9,11-CLA via the ∆9-desat-
serve as a “paracrine factor” in regulating the growth urase reaction. Studies evaluating the feasibility of
of mammary epithelial cells. In the rat mammary using vaccenic acid as a precursor for the endogenous
epithelium, there are morphologically distinctive synthesis of 9,11-CLA in achieving cancer protection
structures called terminal end buds (TEBs) which are will be described. The desaturation and elongation of
present at the tip of some subtending tubules of the CLA in animal tissues have been well documented. This
mammary tree. TEBs are the primary sites for the knowledge opens up a new avenue of research which is
chemical induction of mammary carcinomas. We will related to the question of whether the metabolism of
present data showing that CLA is able to inhibit the CLA is essential for its anticancer activity. For scientific
formation of premalignant lesions from TEBs after reasons, it is critical to delineate whether CLA or one
exposure to a carcinogen. Clonal expansion of an early of its metabolites, is the proximate effector molecule.
transformed pathology is the net result of cell prolifer- Future research direction needs to focus on the
ation minus cell death. Both of these pathways are signaling pathway of CLA and the molecular targets
regulated by a large number of genes whose protein that are responsible for the anticancer effect of CLA.
products act as molecular switches in either a positive
or negative manner. We will discuss some of our recent
PERSPECTIVES ON
16
Conjugated Linoleic Acid Research
CLA Modulation of Mammary Stromal Margot M. Ip and Patricia A. Masso-Welch
Differentiation Contributes to It’s
Chemopreventive Activity Roswell Park Cancer Institute
Buffalo, New York
CLA has been shown to have marked chemopreventive specificity of this response. Significantly, concurrent
activity in rat mammary carcinogenesis models. In part, with MSC differentiation along the adipogenic lineage,
CLA exerts this effect by acting directly on the mamma- there was a decreased ability of MSC to form micro-
ry epithelium to inhibit DNA synthesis and stimulate capillary networks in vitro on an EHS tumor-derived
apoptosis. The objective of our current studies has reconstituted basement membrane (RBM). This
been to determine if CLA might also act indirectly, suggested that CLA might inhibit angiogenesis in vivo.
by modifying the mammary stroma. To examine this, To test this, mice were fed diets with or without CLA
we investigated the effect of CLA on a multipotent for 6 weeks, and then injected subcutaneously with
stromal-vascular cell (MSC) population which is present an angiogenic gel substrate composed of RBM
in the rat mammary gland, and which is able to acquire supplemented with βFGF and heparan sulfate. One
a fibroblastic, adipocyte or endothelial phenotype, week later, the RBM pellets were harvested and
depending on culture conditions (Zangani et al, examined histologically. These studies demonstrated
Differentiation 64: 91, 1999). In these experiments, that functional angiogenesis (formation of red blood
t10,c12-CLA was found to be a potent adipogenic cell-containing vessels) was decreased by ~80%. CLA
factor, stimulating MSC to the adipogenic differentia- also significantly decreased serum and mammary gland
tion pathway even in the absence of exogenous concentrations of vascular endothelial growth factor
hormonal supplementation; c9,t11-CLA was less (VEGF), and the mammary gland VEGF receptor, flk-1.
effective. This effect of CLA was accompanied by a In summary, the ability of CLA to modulate mammary
rapid loss in the DNA-binding activity of the stromal cell differentiation and decrease angiogenesis
PPARγ/RXRα heterodimeric transcription factor may contribute to its efficacy in inhibiting mammary
complex, suggesting that PPARγ may play a key role carcinogenesis.
in initiating the recruitment of MSC into the adipogenic
pathway. DNA-binding activity of other transcription
factors examined was not decreased, demonstrating the
PERSPECTIVES ON
17
Conjugated Linoleic Acid Research
CLA in Experimental Atherosclerosis D. Kritchevsky
The effect of dietary CLA on experimental atherosclero- When 1% CLA was fed to rabbits bearing pre-estab-
sis has been studied in hamsters and rabbits. Hamsters lished atherosclerosis it led to a significant regression
fed 0.12% cholesterol and 1% CLA had significantly of the pre-established lesions. In one study regression
lower plasma cholesterol levels than controls and amounted to 31%, vs. 2% regression in the controls.
exhibited significantly less severe aortic sudanophilia. In a second study the respective values for CLA and
In rabbits fed a semipurified diet containing 0.2% controls were -30% severity and +8% severity. Lower
cholesterol for 90 days, 1% dietary CLA inhibited levels of dietary CLA were without effect in the regres-
atherosclerosis by 36 and 58% respectively in two sion experiments. A study of individual CLA isomers'
experiments. Lower concentrations of dietary CLA also effects on regression is in progress,
reduced severity of atherosclerosis. In one study, CLA
at 0.1 or 0.5% of the diet reduced severity by 34 and
64% respectively. An anti-atherogenic effect has been
observed in rabbits fed as little as 0.05% CLA. These
studies were conducted using a mixture of the major
CLA isomers (about 42-44% each of the c9,t11 and
t10,c12 modifications). The individual isomers (fed as
0.5% of the diet) each have about the same effect
on atherogenicity as does the mixture.
PERSPECTIVES ON
18
Conjugated Linoleic Acid Research
Conjugated Linoleic Acid’s (CLA) Role in Immunity Mark E. Cook, Mingder Yang, Leah Whigham,
and Immune Related Disorders Dan Butz, Guangming Li
PERSPECTIVES ON
19
Conjugated Linoleic Acid Research
Conjugated Linoleic Acid Reduces Fasting Glucose Martha A. Belury, Ph.D., R.D.
and is Inversely Correlated with Serum Leptin in
Subjects with Type 2 Diabetes Mellitus Department of Molecular Medicine, Northwest Hospital
Seattle, Washington
Conjugated linoleic acid (CLA) delays the onset of (r = - 0.4314; P<0.050). Because it appears that
diabetes in the Zucker diabetic fatty (ZDF; fa/fa) rats individual isomers of CLA may differentially alter body
(Biochem Biophys Res Comm 244: 678-682, 1998). composition of experimental animals, we determined
In addition to normalizing impaired glucose in an oral the relationship of the naturally occurring isomer
glucose tolerance test, CLA (1.5wt%) significantly of CLA in the diet, c9t11-CLA (or rumenic acid),
reduced epidydimal fat mass and serum leptin levels. to changes in body weight and serum leptin. In
The data suggested that CLA was able to delay diabetes comparison to correlation coefficients for total plasma
through a mechanism targeting adipose tissue in CLA to a change in body weight or serum leptin,
this experimental animal model. The objective of the correlation coefficients of the level of rumenic acid
present study was to elucidate the relationship of (c9t11-CLA) in plasma were reduced for body weight
supplemental CLA to improvements in the management (r= - 0.3230, P<0.200) and serum leptin (r = - 0.3961;
of type 2 diabetes mellitus. We conducted a double- P<0.100). These findings indirectly suggest the altern-
blind randomized study in subjects with type 2 dia- ative isomer, t10c12-CLA, may exert a more potent
betes supplemented with CLA (8.0 g, 76% pure CLA; effect than c9t11-CLA on reducing body weight and
n= 11) or placebo (8.0 g safflower oil, n=10) daily serum leptin in subjects with type 2 diabetes. Because
for eight weeks. The supplements were 76% CLA the reduced body weights were significantly correlated
containing approximately 37% c9t11-CLA and 39% with reduced fasting blood glucose levels (r = 0.4601;
t10c12-CLA. Dietary assessment of intake of energy or P<0.050), our study suggests the improvement in fast-
fat composition revealed no differences at baseline or ing blood glucose by supplemental CLA may occur
week 8 for either treatment group. Supplementation through lowering body weights and/or altering body
with CLA significantly decreased fasting blood glucose composition. Further work is needed to identify the
(P< 0.050) and exerted a modest trend for decreasing role of CLA in improving insulin sensitivity, reducing
fasting plasma insulin (p <0.100). The strengths of the body weight and altering mass and distribution of
associations of plasma levels of CLA to changes in body adipose tissue in humans. In addition, future studies
weight and serum leptin were determined by quantify- should determine the optimal doses and isomeric
ing correlation coefficients. Plasma CLA was inversely mixtures of CLA required to aid in the management
correlated, although not significantly, with a change of type 2 diabetes mellitus in a longterm study.
in body weight (r = - 0.3739; P<0.100) and significant-
ly inversely correlated with a change in leptin
PERSPECTIVES ON
20
Conjugated Linoleic Acid Research
CLA and Bone Formation Bruce A. Watkins
Skeletal metabolism is controlled by cells of the bone (20:4n-6) was decreased in liver polar lipids by CLA but
and joint microenvironments through the actions of not in bone. In the neutral lipid fraction of most rat
prostaglandins, cytokines, and growth factors involved tissues analyzed, CLA treatment decreased 18:1, 20:2,
in the local regulation of bone metabolism. New 20:4n-6, 22:5n-3, 22:6n-3, total monounsaturated,
studies suggest that specific PUFA improve bone total n-6, total n-3, and total PUFA, but increased
metabolism and reduce or control the risk for saturated fatty acids. Rat serum osteocalcin level and
bone/joint diseases. The PUFA and to some extent bone specific alkaline phosphatase (BALP) activity
conjugated linoleic acid (CLA) modulate eicosanoid was decreased in rats fed CLA. In contrast, rats given
biosynthesis in osteoblasts, alter biomarkers of bone the diet containing a moderate level of n-6 PUFA
formation, impact bone formation rates in rats, and relative to the high n-6 PUFA had a higher rate of
influence gene expression during osteoblast maturation bone formation in the tibia. In addition, the supple-
and matrix formation. The first published study on ment of CLA appeared to be protective in supporting
CLA and bone formation showed that 1% dietary CLA bone formation in rats given a higher level of n-6
isomers depressed ex vivo PGE2 production in rat bone PUFA. Studies in osteoblasts enriched with CLA isomers
organ culture, reduced serum IGF-I, and reduced bone during proliferation, maturation, and mineralization
formation rate in rat long bone. These responses were indicate the CLA down-regulates COX enzymes and
influenced by the dietary ratio of n-6/n-3 fatty acids. has variable effects on signaling proteins and gene
In a subsequent study, a lower dietary level (0.5%) of expression. In other experiments, bone mineral
CLA was supplemented to diets containing moderate or content and bone mineral density measured by DEXA
high levels of PUFA (moderate or high n-6 PUFA oil in ovariectomized rats was not improved by CLA
blend) appeared to rescue bone formation rate in male supplementation alone. Our research on CLA isomers
rats. The dietary lipid treatments did not affect growth; in rats and other mammals indicates that the actions
however, CLA improved feed efficiency during the first of these isomers is dependent on the type of dietary
six weeks of feeding. CLA isomers were found in all rat fat, the balance of PUFA (dietary ratio of n-6/n-3
tissues analyzed and CLA content in neutral lipid was 5 fatty acids), and may influence factors at the
to 10 times greater than that in the polar fraction. CLA molecular level.
lowered 18:1n-9 and total monounsaturated fatty acids
while it increased 22:6n-3 and total n-3 in the polar
fraction of liver and bone marrow. Arachidonic acid
PERSPECTIVES ON
21
Conjugated Linoleic Acid Research
Nutritional Regulation of Josep Bassaganya-Riera
Bacterial-Induced Colitis by
Conjugated Linoleic Acid Veterinary Medical Research Institute
Iowa State University, Ames, Iowa
Excessive intake of saturated fatty acids and/or linoleic pathogen (i.e., Brachyspira hyodysenteriae).
acid favors the induction of an array of lipid mediators Immunoregulatory cytokines as well as peroxisome-
and cytokines enhancing inflammatory responses. proliferator activated receptor-γ (PPAR-γ) mRNA
Conversely, dietary supplementation with n-3 fatty expression was assayed in colonic lymph nodes and
acids or vitamin D ameliorates inflammation and colon of pigs. Colonic mucosal lesions and lymphocyte
autoimmune diseases. While it was well-accepted subset distribution were evaluated by histology and
that conjugated linoleic acid (CLA) prevented diseases immunohistochemistry. Supplementation of CLA in
with a common inflammatory pathogenesis (i.e., the diet prior to the induction of colitis decreased
cancer, diabetes, and atherosclerosis), no studies were mucosal damage, maintained cytokine profiles (i.e.,
available on the roles of CLA on mucosal inflammation. interferon-γ and interleukin-10) and lymphocyte
The present study aimed at investigating the anti- subset distributions (i.e., CD4+ and CD8+) resembling
inflammatory actions and molecular mechanisms those of non-infected pigs, enhanced colonic
underlying the regulation of colonic health by CLA. expression of PPAR-γ and attenuated growth failure.
It was hypothesized that colonic inflammation can be Therefore, CLA fed preventively prior to the onset
ameliorated by dietary CLA supplementation. To test of enteric disease attenuated inflammatory lesion
this hypothesis, inflammation of the colonic mucosa development and growth failure.
was triggered by challenging pigs fed either soybean
oil or CLA-supplemented diets with an enteric bacterial
PERSPECTIVES ON
22
Conjugated Linoleic Acid Research
Effects of CLA in Obese Subjects Richard L. Atkinson, M.D.
on a Weight Loss Diet: Wisconsin Data
Departments of Medicine and Nutritional Sciences,
University of Wisconsin, Madison, Wisconsin
Obesity is a chronic disease that is resistant to times weekly. Body composition was assessed by
diet, exercise, and lifestyle modification treatments. underwater weighing. 71 subjects (41 F, 30 M) finished
Pharmacologic treatment is somewhat more successful, the 6 mo trial. CLA subjects lost 2.4 kg vs 2.2 kg for
but safety and long term efficacy are not clear. Drug placebo. Fat mass declined by 1.3 kg and 1.0 kg,
treatment must be long term because cessation of respectively. Fat free mass decreased by 1.1 kg and
treatment invariably leads to weight regain. Conjugated 1.2 kg, respectively. Laboratory variables did not differ
linoleic acid (CLA) in growing animals reduces body between the groups. Side effects and adverse events
fat and increases lean body mass vs control animals. were significantly fewer in the CLA group (p<.05).
Human trials show no effect or modest reduction in We conclude that CLA does not enhance weight loss
body weight or body fat with CLA compared to placebo. or reduce body fat in obese subjects on a weight
We did a randomized, double-blind, placebo-controlled loss program, but that it appears to be safe and to
trial in 80 obese subjects treated for 6 months with reduce side effects during weight loss over 6 months.
placebo or 2.7 gm of CLA/day. Characteristics were Additional studies are needed in humans with research
mean age 41.5 yr, mean wt 94.0 kg, and absence of designs comparable to the animal studies to determine
severe illness, pregnancy, lactation, or interfering if CLA prevents adipose tissue accumulation.
drugs. Subjects were asked to reduce customary intake
by 500 kcal/d and to exercise for 30 min at least 3
PERSPECTIVES ON
23
Conjugated Linoleic Acid Research
Clinical Studies on Metabolic Effects of B. Vessby, U. Risérus, A. Smedman and S. Basu
Conjugated Linoleic Acid in Humans
Unit for Clinical Nutrition Research, Department of
Public Health and Caring Sciences, University of Uppsala,
Uppsala, Sweden
Conjugated linoleic acid (CLA) comprises a group markers of lipid peroxidation and on endocrine and
of unsaturated fatty acid isomers with a variety of proinflammatory factors. Preliminary results indicate
biological effects in experiental animal studies. CLA that CLA may slightly decrease body fat also in humans,
reduces body fat accumulation and has been ascribed particularly abdominal fat, but there is no simultaneous
significant effects on lipid and glucose metabolism, improvement of lipid or glucose metabolism. Rather,
e.g. antidiabetic effects in obese Zucker rats. It has the t10c12 isomer unexpectedly caused significant
been suggested that the t10c12 CLA isomer is the impairment of the peripheral insulin sensitivity as well
active isomer as regards antiobesity and insulin as of blood glucose and serum lipid levels. In addition,
sensitizing properties of CLA. The metabolic effects of CLA markedly elevated lipid peroxidation. Thus, the
CLA in humans in general, and isomer specific effects metabolic effects of CLA in humans seem complex and
in particular, are not well characterized. We have in a further studies, especially of isomer specific effects,
series of controlled studies in humans investigated are needed.
the effects of CLA (given as the commercially availabe
mixture of isomers) and of the purified t10c12 isomer
on anthropometry, lipid and glucose metabolism, on
PERSPECTIVES ON
24
Conjugated Linoleic Acid Research
Seroprotection: CLA Stimulates Antigen Marianne O’ Shea,1 Ruud Albers,3 Reggy van der Wielen,3 Lisette
Brink,4 V. Dorovska-Taran,2 Inge Mohede2
Specific Antibody Production in Humans
1 Loders Croklaan, Lipid Nutrition, Channahon, Illinois
2 Loders Croklaan, Lipid Nutrition, Wormerveer, The Netherlands
3 Unilever Health Institute, Vlaardingen, The Netherlands
4 TNO Nutrition and food research institute, Zeist, The Netherlands
Considerable evidence exists that CLA enhances immune (SPR, i.e. the number of subjects with anti-Hbs concen-
function in vitro- and in animal-studies. In this study, trations >10 IU/L compared to the number of subjects
the potential of CLA to modulate the human immune with titers <10 IU/L) was significantly higher (P=0.05)
system was investigated using the two main isomers for the 50:50 group compared with the control or the
in different ratios (50:50 and 80:20 of c9,t11:t10,c12 80:20 group. The cell mediated immune response was
CLA, respectively). The humoral and cell mediated measured using the CMI multitest for “Delayed-Type
immune responses were investigated in humans Hypersensitivity” (DTH). Evaluation of the DTH
supplemented with CLA (1.7g active isomers/day for responses on 7 recall antigens, at different time
12 weeks). points showed no statistically significant differences
in all groups.
Hepatitis B (Hbs) vaccination was used as an infection
model to investigate the humoral and cell mediated This is the first study in humans that clearly demon-
immune response. Hepatitis B antibody titres were strates stimulation of the humoral immune (antibody)
evaluated for each subject on day 0 and 2 weeks post response by CLA supplementation as reflected by an
initial vaccination and final booster. Mean serum Hbs increase of the SPR.
antibody concentration at day 85 was twice as high
for subjects consuming CLA 50:50 compared with the
control or the 80:20 group. The seroprotection rate
PERSPECTIVES ON
25
Conjugated Linoleic Acid Research
Chairpersons
Chairpersons
PERSPECTIVES ON
29
Conjugated Linoleic Acid Research
Speakers
Speakers
PERSPECTIVES ON
32
Conjugated Linoleic Acid Research
Speakers
PERSPECTIVES ON
33
Conjugated Linoleic Acid Research
Speakers
PERSPECTIVES ON
34
Conjugated Linoleic Acid Research
Speakers
PERSPECTIVES ON
35
Conjugated Linoleic Acid Research
Participants
Participants
Mel Dong
5100 Paint Branch Parkway, HFS-255
College Park, MD 20740
Phone: (202) 418-3048
wdong@cfsan.fda.gov
PERSPECTIVES ON
38
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
39
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
40
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
41
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
42
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
43
Conjugated Linoleic Acid Research
Participants
PERSPECTIVES ON
44
Conjugated Linoleic Acid Research
Participants
Jaimebeth Vicidomini
6111 Borken Sound Parkway
Boca Raton, FL 33487
Phone: (561) 999-1334
jvicidomini@rexallsundown.com
PERSPECTIVES ON
45
Conjugated Linoleic Acid Research
Notes
PERSPECTIVES ON
46
Conjugated Linoleic Acid Research
Notes
PERSPECTIVES ON
47
Conjugated Linoleic Acid Research
Notes
PERSPECTIVES ON
48
Conjugated Linoleic Acid Research
Notes
PERSPECTIVES ON
49
Conjugated Linoleic Acid Research