Hepatitis C viraemia is carbohydrate-dependent because the virus piggy-backs on triglyceride assembly and VLDL exocytosis. This makes a very low carbohydrate diet an effective way to control HCV viraemia, HCV-associated autoimmune syndromes, and steatosis. HCV cell entry is via LDL-receptor complex, therefore diets intended to lower LDL via upregulation of the LDL-receptor by restricting saturated fat and increasing polyunsaturated fat will increase hepatocellular infection.
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Thursday, 7 April 2016
On second thoughts, that vegetarian genomic study did show that not eating animals is not good for you.
Generally, a study that purported to show that vegan and vegetarian diets are harmful would be welcomed by meat eaters, who get a lot of pseudoscientific criticism from members of those groups, some of it disguised as sober science.
But no-one was much impressed by the Pune vs. Kansas study. Even Tom Naughton wrote it off as meaning the same thing the head of the NZ vegetarian society said it meant - that omega-6 seed oils just aren't good for us anyway.
But I thought about this, and, not so fast.
Seed oils high in omega 6 are harmful for the descendants of long lines of vegetarians because such people, because of an adaptation to the virtual absence, from their diets, of DHA and AA (arachidonic acid), the very long-chain PUFAs found in animal flesh and organ meats, have a more efficient version of the genes involved in synthesizing these fats from alpha-linolenic acid (ALA) and linoleic acid (LA). Too much LA overwhelms these enzymes, which only seem to be loosely regulated, and results in an excess of inflammatory AA products and an inadequacy of very long chain omega 3s.
So this adaptation is good for vegetarians eating traditional diets, as in Pune where the traditional fat source would have been ghee, with a little mustard seed oil added. Low in omega 6, balanced in omega 3, enough hearty saturated dairy fat to protect against the diabetogenic effect of a diet high in both starch and sugar.
But think about it - this adaptation isn't some random lucky fluke. For one gene to dominate over another like this, there needs to be some significant and sustained reproductive advantage.
Reproductive advantage means one or more of these - greater fertility, fewer stillbirths, fewer complications of pregnancy, lower mortality early in life, greater attractiveness to a mate.
The vegetarian PUFA polymorphism flourished because, in the past, people without it, eating vegetarian diets, suffered some combination of infertility, stillbirth, dangerous pregnancy, early mortality, or plain butt-ugliness.
Its incidence at present is 70% in South Asians, 53% in Africans, 29% in East Asians, and 17% in Europeans. That to me indicates a burden of suffering and infertility in South Asians in the past, to produce this result - that's how evolution works, that's how Nature selects. If you're European, the chances are that you do need AA and DHA in your food, unless you want to take your chances with lots of vegetable oil - which seems to me a very second-rate, artificial, and dicey way of getting there.
Note that some vegans do think it's okay to eat bivalve shellfish, which can't feel pain (or rather, probably don't feel more pain that plants do, but who knows what that is). This would supply more than enough DHA and AA. However, PETA takes the hard line on this, like the Buddhist who won't swat a zika-carrying mosquito.
But then, PETA is Neal Barnard's baby and he's a dietary cholesterol zealot, so their ban on shellfish might not be as strictly ethical as they claim. Dr Barnard "advises people to avoid added vegetable oils and other high-fat foods as well as refined sugar and flour". Well good for him but it is hard to see where the AA and DHA will come from for the majority of Europeans on this diet.
Maybe veganism is a bit like statinism - enough of the people it's going to harm will drop out of the trial early for the long-term results to look a bit encouraging. It would be interesting to see if long-term vegans in European populations have in fact self-selected for the FADS2 polymorphism common in Pune.
Thursday, 31 March 2016
Silymarin for type 2 diabetes - significant effects on glucose and lipids from a safe OTC herbal.
This study has an interesting backstory.
Hepatitis C (mainly genotype 4) infects nearly a quarter of the Egyptian population. This is the highest rate of HCV infection I've heard of in any country; however the Nile Valley is probably the ancestral home of HCV's transmission to humans.
Egypt is not a rich country and drug treatments for Hep C are expensive, not to mention dangerous and unreliably effective till recently. Consequently a lot of Egyptians use alternative remedies, usually sourced from EU pharmacopoeias. Silymarin (a standardised mik thistle extract) and a German spirulina extract are two of the most popular; I wrote some time ago about their relative effect on hepatitis C infection.
Edit - the spirulina and silymarin in that earlier study was supplied by Beovita-Safe Pharma, a Joint German Egyptian Company, Katzbachstr. 29, D-10965 Berlin. There is no mention of the supplier of silymarin in the latest study, but it may be from the same source.
These remedies are so widely used in Egypt that Egyptian pharmacologists have investigated their safety and effectiveness with unusual thoroughness. It's not a big leap from treating the fatty liver of chronic hep C infection to seeing if silymarin will improve type 2 diabetes. This is a disease highly associated with NAFLD, and abnormal liver function is thought to be a primary cause of diabetic insulin resistance and dyslipidemia.
Hepatitis C (mainly genotype 4) infects nearly a quarter of the Egyptian population. This is the highest rate of HCV infection I've heard of in any country; however the Nile Valley is probably the ancestral home of HCV's transmission to humans.
Egypt is not a rich country and drug treatments for Hep C are expensive, not to mention dangerous and unreliably effective till recently. Consequently a lot of Egyptians use alternative remedies, usually sourced from EU pharmacopoeias. Silymarin (a standardised mik thistle extract) and a German spirulina extract are two of the most popular; I wrote some time ago about their relative effect on hepatitis C infection.
Edit - the spirulina and silymarin in that earlier study was supplied by Beovita-Safe Pharma, a Joint German Egyptian Company, Katzbachstr. 29, D-10965 Berlin. There is no mention of the supplier of silymarin in the latest study, but it may be from the same source.
These remedies are so widely used in Egypt that Egyptian pharmacologists have investigated their safety and effectiveness with unusual thoroughness. It's not a big leap from treating the fatty liver of chronic hep C infection to seeing if silymarin will improve type 2 diabetes. This is a disease highly associated with NAFLD, and abnormal liver function is thought to be a primary cause of diabetic insulin resistance and dyslipidemia.
Effect of Silymarin Supplementation on
Glycemic Control, Lipid Profile and Insulin Resistance in Patients with Type 2
Diabetes Mellitus. (full text here)
Amany Talaat Elgarf 1,
Maram Maher Mahdy 2, Nagwa Ali Sabri 1
International Journal of Advanced Research (2015), Volume 3, Issue 12, 812 – 821.
1. Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt. 2. Department of Internal Medicine and Diabetes, Faculty of Medicine, Ain Shams University, Cairo, Egypt
Note that Ain Shams is a proper medical school, the 3rd oldest in Egypt, founded in 1950.
Forty patients were randomly assigned to receive either silymarin capsules 140 mg three times daily (n=20) or identical placebo capsules three times daily (n=20) for 90 days. Full clinical history and fasting blood samples were obtained to determine FBG , HbA1c, FSI, full lipid profile, MDA , hs-CRP levels as well as HOMA-IR at the beginning and at the end of the study.
These results are pretty impressive. Firstly, the control group is getting worse in every parameter tested over the study period, and many of the differences are significant.
Meanwhile, the silymarin arm sees some striking improvements. The authors highlight a rise in HDL from 23 (CI 12.0 - 52.0) to 38.5 (CI 14.0 - 65.0) mg/dl, which is consistent with an improvement in HOMA-IR and a drop in fasting insulin from 15.2 (8.4-20.7) to 11.2 (9.3-15.6) uIU/mL. Over the same 3 months insulin rose to 19.7 (9.4-24.4) uIU/mL in the placebo group.
Also impressive is the drop in LDL-C and LDL-C. LDL-C drops from 131.9 (69.0-218.6) to 94.0 (58.8-154.2) mg/dl, and VLDL-C drops from 34.3 (19.0-47.0) to 20.8 (16.6-35.0) mg/dl.
Remember that a diagnosis of diabetes is one of the criteria for prescribing statins. Statins can lower LDL-C, but they won't lower blood glucose, in fact they double the chance of it rising into the diabetic range. Silymarin, on the other hand, lowered fasting BG from 252.5 (174.0-395.0) to 162.0 (109.0-391.0) mg/dl (while it rose 20% in the placebo arm during the same period). HbA1c dropped from 10.4 (8.0-12.3) to 8.5 (6.3-12.3) %.
Basically, a safe OTC supplement seems to be able to give the benefits of metformin and statins combined, with a minimal risk. The safety of silymarin is recorded in dozens of long term Hep C studies of various types.
Would silymarin have benefits for people on low-carb diets who see a large rise in LDL, or whose blood glucose control still isn't perfect? I think it might be worth trying.
International Journal of Advanced Research (2015), Volume 3, Issue 12, 812 – 821.
1. Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt. 2. Department of Internal Medicine and Diabetes, Faculty of Medicine, Ain Shams University, Cairo, Egypt
Note that Ain Shams is a proper medical school, the 3rd oldest in Egypt, founded in 1950.
Forty patients were randomly assigned to receive either silymarin capsules 140 mg three times daily (n=20) or identical placebo capsules three times daily (n=20) for 90 days. Full clinical history and fasting blood samples were obtained to determine FBG , HbA1c, FSI, full lipid profile, MDA , hs-CRP levels as well as HOMA-IR at the beginning and at the end of the study.

These results are pretty impressive. Firstly, the control group is getting worse in every parameter tested over the study period, and many of the differences are significant.
Meanwhile, the silymarin arm sees some striking improvements. The authors highlight a rise in HDL from 23 (CI 12.0 - 52.0) to 38.5 (CI 14.0 - 65.0) mg/dl, which is consistent with an improvement in HOMA-IR and a drop in fasting insulin from 15.2 (8.4-20.7) to 11.2 (9.3-15.6) uIU/mL. Over the same 3 months insulin rose to 19.7 (9.4-24.4) uIU/mL in the placebo group.

Also impressive is the drop in LDL-C and LDL-C. LDL-C drops from 131.9 (69.0-218.6) to 94.0 (58.8-154.2) mg/dl, and VLDL-C drops from 34.3 (19.0-47.0) to 20.8 (16.6-35.0) mg/dl.
Remember that a diagnosis of diabetes is one of the criteria for prescribing statins. Statins can lower LDL-C, but they won't lower blood glucose, in fact they double the chance of it rising into the diabetic range. Silymarin, on the other hand, lowered fasting BG from 252.5 (174.0-395.0) to 162.0 (109.0-391.0) mg/dl (while it rose 20% in the placebo arm during the same period). HbA1c dropped from 10.4 (8.0-12.3) to 8.5 (6.3-12.3) %.
Basically, a safe OTC supplement seems to be able to give the benefits of metformin and statins combined, with a minimal risk. The safety of silymarin is recorded in dozens of long term Hep C studies of various types.
Would silymarin have benefits for people on low-carb diets who see a large rise in LDL, or whose blood glucose control still isn't perfect? I think it might be worth trying.
Thursday, 24 March 2016
The Smoking Gun - the Role of PUFA in Non-Alcoholic Liver Disease
The smoking gun
Public health experts are gradually
accepting the idea that sucrose and fructose are, like alcohol, causes of fatty
liver disease (non-alcoholic liver disease - NAFLD - and its inflammatory
development, non-alcoholic steatohepatitis - NASH).
After all, sugar is unnecessary and, like alcohol, the rogue macronutrient, associated with pleasure rather than nutrition. There’s little or no evidence that there is ever likely to be a health benefit from replacing starch or fat with sugar.
Sugar was first equated with alcohol in a liver disease model by CH Best, co-discoverer of insulin, in 1949,[1] a fact which has a nice aptness to it, because NAFLD is often the first stage that leads to type 2 diabetes and, if you’re not very careful about the quality of food and the calories and carbs, insulin-dependence.
On the other hand, there is little mainstream acceptance of the idea that polyunsaturated fat plays a role in these diseases, with the honorable exception of Canada’s recent obesity report; yet the scientific evidence that dietary fats of 5% or more PUFA are essential for the development of alcoholic liver disease (ALD) is very strong. (See here and here)
Polyunsaturated fat is the Golden Boy of public health – seed oils have saved the world from heart disease, supposedly, so the public presentation of evidence that they promote other diseases has always faced an uphill battle.
For a start, PUFA is a small part of the diet and isn’t measured with great accuracy in epidemiological studies. Its harms are interactive with two other nutrients – sugars and alcohol – the excess consumption of which may not be reported as accurately or honestly as intake of other foods.
Anyway, this new study tells us that the genes that encode proteins (enzymes) needed for the metabolism and detoxification of alcohol are upregulated in NAFLD. I can’t get full-text for this, but the abstract is informative.
After all, sugar is unnecessary and, like alcohol, the rogue macronutrient, associated with pleasure rather than nutrition. There’s little or no evidence that there is ever likely to be a health benefit from replacing starch or fat with sugar.
Sugar was first equated with alcohol in a liver disease model by CH Best, co-discoverer of insulin, in 1949,[1] a fact which has a nice aptness to it, because NAFLD is often the first stage that leads to type 2 diabetes and, if you’re not very careful about the quality of food and the calories and carbs, insulin-dependence.
On the other hand, there is little mainstream acceptance of the idea that polyunsaturated fat plays a role in these diseases, with the honorable exception of Canada’s recent obesity report; yet the scientific evidence that dietary fats of 5% or more PUFA are essential for the development of alcoholic liver disease (ALD) is very strong. (See here and here)
Polyunsaturated fat is the Golden Boy of public health – seed oils have saved the world from heart disease, supposedly, so the public presentation of evidence that they promote other diseases has always faced an uphill battle.
For a start, PUFA is a small part of the diet and isn’t measured with great accuracy in epidemiological studies. Its harms are interactive with two other nutrients – sugars and alcohol – the excess consumption of which may not be reported as accurately or honestly as intake of other foods.
Anyway, this new study tells us that the genes that encode proteins (enzymes) needed for the metabolism and detoxification of alcohol are upregulated in NAFLD. I can’t get full-text for this, but the abstract is informative.
“Alcohol-metabolizing enzymes including
ADH, ALDH, CYP2E1, and CAT were up-regulated in NAFLD livers. The expression
level of alcohol-metabolizing genes in severe NAFLD was similar to that in AH.”
“[I]ncreased expression of alcohol-metabolizing genes in NAFLD livers supports a role for endogenous alcohol metabolism in NAFLD pathology and provides further support for gut microbiome therapy in NAFLD management.”[2]
Well yes, there is definitely a role for probiotics and prebiotics (which now include long-chain saturated fats) in NAFLD and ALD management. But the idea that NAFLD is caused by endogenous alcohol production in all but a few cases seems preposterous to me. Alcoholic liver disease is associated with drunkenness, alcoholism, and thiamine depletion. Are these seen in patients with NAFLD?
However, was alcohol involved, there would be the same disease-promoting role for PUFA seen in ALD.
Why else would alcohol-metabolising enzymes be upregulated? We didn’t evolve drinking alcohol, so why did this enzyme system come to exist?
It exists originally for the metabolism of polyunsaturated fats into eicosanoids, that is to say, into inflammatory molecular messengers, and for the removal of oxidised PUFAs.
For example, if you feed oxidized linoleic acid to rats, their expression of aldehyde dehydrogenase (ALDH) increases.[3] The alcohol dehydrogenase (ADH) enzyme in leeks breaks down essential fatty acids into aromatic metabolites (sure, a leek isn’t a human, but it shows that ADH enzymes act on PUFAs in the absence of alcohol, which is what we want to know). [4]And if you feed PUFAs to cultured hepatoma (HepG2) cells, which is the cell culture model for liver diseases, you get this:
“After 2 hours of cultivation, the lipid peroxide (LPO) in the DHA group increased 600% compared with control, and was much higher than in the groups treated with the other FAs, with LNA > LA > OA > PA. CYP2E1 induction increased with greater effect as the degree of unsaturation of OA, LA, and DHA increased.”[5]
PA was palmitic acid, and had no effect on PKC activity, the marker of cellular stress in the experiment.
CAT is catalase, a heme enzyme which degrades H202 to water and oxygen, the end of this detox disassembly line.
“The effects of linoleic and intake on catalase and other enzymes were investigated by feeding 0, 1, 5 or 10% corn oil diet to rats previously fed a fat-free diet. Rats fed more than 1% corn oil for 2 weeks showed significant increases of glutathione peroxidase and superoxide dismutase in liver cytosol when compared to the controls fed no corn oil. Peroxisomal catalase activity especially was increased.”[6]
“[I]ncreased expression of alcohol-metabolizing genes in NAFLD livers supports a role for endogenous alcohol metabolism in NAFLD pathology and provides further support for gut microbiome therapy in NAFLD management.”[2]
Well yes, there is definitely a role for probiotics and prebiotics (which now include long-chain saturated fats) in NAFLD and ALD management. But the idea that NAFLD is caused by endogenous alcohol production in all but a few cases seems preposterous to me. Alcoholic liver disease is associated with drunkenness, alcoholism, and thiamine depletion. Are these seen in patients with NAFLD?
However, was alcohol involved, there would be the same disease-promoting role for PUFA seen in ALD.
Why else would alcohol-metabolising enzymes be upregulated? We didn’t evolve drinking alcohol, so why did this enzyme system come to exist?
It exists originally for the metabolism of polyunsaturated fats into eicosanoids, that is to say, into inflammatory molecular messengers, and for the removal of oxidised PUFAs.
For example, if you feed oxidized linoleic acid to rats, their expression of aldehyde dehydrogenase (ALDH) increases.[3] The alcohol dehydrogenase (ADH) enzyme in leeks breaks down essential fatty acids into aromatic metabolites (sure, a leek isn’t a human, but it shows that ADH enzymes act on PUFAs in the absence of alcohol, which is what we want to know). [4]And if you feed PUFAs to cultured hepatoma (HepG2) cells, which is the cell culture model for liver diseases, you get this:
“After 2 hours of cultivation, the lipid peroxide (LPO) in the DHA group increased 600% compared with control, and was much higher than in the groups treated with the other FAs, with LNA > LA > OA > PA. CYP2E1 induction increased with greater effect as the degree of unsaturation of OA, LA, and DHA increased.”[5]
PA was palmitic acid, and had no effect on PKC activity, the marker of cellular stress in the experiment.
CAT is catalase, a heme enzyme which degrades H202 to water and oxygen, the end of this detox disassembly line.
“The effects of linoleic and intake on catalase and other enzymes were investigated by feeding 0, 1, 5 or 10% corn oil diet to rats previously fed a fat-free diet. Rats fed more than 1% corn oil for 2 weeks showed significant increases of glutathione peroxidase and superoxide dismutase in liver cytosol when compared to the controls fed no corn oil. Peroxisomal catalase activity especially was increased.”[6]
So, with a very cursory search, I found
that the 4 enzymes found upregulated in ref. [2] metabolise PUFAs, and are upregulated
when they are present in quantity.
No endogenous alcoholism is needed to explain this result.
The next question – how does the presence of excess fructose drive this enzyme system? Alcohol upregulates the enzyme system because it degrades alcohol, and PUFA is then caught up in the activated enzymes; but what role does sugar play?
Edit: this is a good place to include recent human evidence for this theory.
5-Hydroxyicosatetraenoic acid (5-HETE) and 9-Hydroxyoctadecadienoic acid (9-HODE) are eicosanoid metabolites of linoleic acid (omega 6 PUFAs). In this Polish study,
" Patients (n=12) with stage I NAFLD had a significantly higher level of HDL cholesterol and a lower level of 5-HETE. Patients (n=12) with grade II steatosis had higher concentrations of 9-HODE. Following the six-month dietary intervention, hepatic steatosis resolved completely in all patients. This resulted in a significant decrease in the concentrations of all eicosanoids (LX4, 16-HETE, 13-HODE, 9-HODE, 15-HETE, 12-HETE, 5-oxoETE, 5-HETE) and key biochemical parameters (BMI, insulin, HOMA-IR, liver enzymes).
Conclusion: A significant reduction in the analyzed eicosanoids and a parallel reduction in fatty liver confirmed the usefulness of HETE and HODE in the assessment of NAFLD."[7]
Steatosis resolved completely after 6 months on a diet in which LA was restricted to 4% of energy and sugar to 10%. Though the diet was low in fat (20-35% of energy) dairy was favoured as a source of fat -
"The type of fat included in the diet was easy to digest, such as cream, butter, oil or milk...The total omega-3 and omega-6 fatty acids consumption was approximately 0.5% E for omega-3 and 4% E for omega-6."
In 2004 the average omega 6 content of the Polish diet was 5.21% "much higher than the recommended upper limit (3% of energy)." (link) As the NAFLD diet was individually calorie-restricted, the total amount of omega-6 would have been close to the total giving the recommended 3% in the normal diet.
We also find reversal of fatty liver disease, associated with obesity and type 2 diabetes, in the recent pilot trial of Unwin et al, where subjects were told to avoid sugar, grains, and other carbohydrate-dense foods.[8]
"In place of carbohydrate-rich foods, an increased intake of green vegetables, whole-fruits, such as blueberries, strawberries, raspberries and the “healthy fats” found in olive oil, butter, eggs, nuts and full-fat plain yoghurt were advocated."
A 50/50 mix of butter and olive oil (for example) gives a fat of around 6% omega 6; nuts and poultry, which are not necessarily foods eaten every day, supply somewhat higher amounts; in the context of a diet around 60-70% fat, these instructions should amount to a high-fat diet that is not excessively high in omega 6; however the effects of carbohydrate restriction on NAFLD are significant even when fat composition is 15% PUFA in a 60% fat, 8% carbohydrate diet, as in the experiment of Browning et al.[9]
These various examples of fatty liver reversal diets seem to indicate the synergy of sugars, carbohydrates, and polyunsaturated fat in the NAFLD dietary model.
No endogenous alcoholism is needed to explain this result.
The next question – how does the presence of excess fructose drive this enzyme system? Alcohol upregulates the enzyme system because it degrades alcohol, and PUFA is then caught up in the activated enzymes; but what role does sugar play?
Edit: this is a good place to include recent human evidence for this theory.
5-Hydroxyicosatetraenoic acid (5-HETE) and 9-Hydroxyoctadecadienoic acid (9-HODE) are eicosanoid metabolites of linoleic acid (omega 6 PUFAs). In this Polish study,
"
Conclusion: A significant reduction in the analyzed eicosanoids and a parallel reduction in fatty liver confirmed the usefulness of HETE and HODE in the assessment of NAFLD."[7]
Steatosis resolved completely after 6 months on a diet in which LA was restricted to 4% of energy and sugar to 10%. Though the diet was low in fat (20-35% of energy) dairy was favoured as a source of fat -
"The type of fat included in the diet was easy to digest, such as cream, butter, oil or milk...The total omega-3 and omega-6 fatty acids consumption was approximately 0.5% E for omega-3 and 4% E for omega-6."
In 2004 the average omega 6 content of the Polish diet was 5.21% "much higher than the recommended upper limit (3% of energy)." (link) As the NAFLD diet was individually calorie-restricted, the total amount of omega-6 would have been close to the total giving the recommended 3% in the normal diet.
We also find reversal of fatty liver disease, associated with obesity and type 2 diabetes, in the recent pilot trial of Unwin et al, where subjects were told to avoid sugar, grains, and other carbohydrate-dense foods.[8]
"In place of carbohydrate-rich foods, an increased intake of green vegetables, whole-fruits, such as blueberries, strawberries, raspberries and the “healthy fats” found in olive oil, butter, eggs, nuts and full-fat plain yoghurt were advocated."
A 50/50 mix of butter and olive oil (for example) gives a fat of around 6% omega 6; nuts and poultry, which are not necessarily foods eaten every day, supply somewhat higher amounts; in the context of a diet around 60-70% fat, these instructions should amount to a high-fat diet that is not excessively high in omega 6; however the effects of carbohydrate restriction on NAFLD are significant even when fat composition is 15% PUFA in a 60% fat, 8% carbohydrate diet, as in the experiment of Browning et al.[9]
These various examples of fatty liver reversal diets seem to indicate the synergy of sugars, carbohydrates, and polyunsaturated fat in the NAFLD dietary model.
[1] C. H. Best, W. Stanley Hartroft, C. C.
Lucas, and Jessie H. Ridout. Liver Damage Produced by Feeding Alcohol or Sugar
and its Prevention by Choline. Br Med J. 1949 Nov 5; 2(4635): [1001]-1004-1,
1005-1006.
[2] Zhu R, Baker SS, Moylan CA, et al. Systematic
transcriptome analysis reveals elevated expression of alcohol-metabolizing
genes in NAFLD livers. The Journal of Pathology Volume 238, Issue 4, pages
531–542, March 2016
[3] Hochgraf E, Mokady S, Cogan U. Dietary Oxidized Linoleic Acid Modifies Lipid Composition of Rat Liver Microsomes and Increases Their Fluidity. J. Nutr. 127: 681–686, 1997.
[3] Hochgraf E, Mokady S, Cogan U. Dietary Oxidized Linoleic Acid Modifies Lipid Composition of Rat Liver Microsomes and Increases Their Fluidity. J. Nutr. 127: 681–686, 1997.
[4] Nielsen GS, Larsen LM, Poll L. Formation
of Volatile Compounds in Model Experiments with Crude Leek (Allium ampeloprasum
Var. Lancelot) Enzyme Extract and Linoleic Acid or Linolenic Acid. J. Agric.
Food Chem. 2004, 52, 2315-2321
[5] Sung M, Kim I. Differential Effects of Dietary Fatty Acids on the Regulation of CYP2E1 and Protein Kinase C in Human Hepatoma HepG2 Cells. J Med Food 7 (2) 2004, 197–203
[6] Iritani N, Ikeda Y. J Nutr. Activation of catalase and other enzymes by corn oil intake. 1982 Dec;112(12):2235-9.
[7] Maciejewska D, Ossowski P, Drozd A, et al. Metabolites of arachidonic acid and linoleic acid in early stages of non-alcoholic fatty liver disease - A pilot study. Prostaglandins Other Lipid Mediat. 2015 Sep;121(Pt B):184-9.
[8] Unwin DJ, Cuthbertson DJ, Feinman R, Sprung VS (2015) A pilot study to explore the role of a low-carbohydrate intervention to improve GGT levels and HbA1c. Diabesity in Practice 4: 102–8.
[9] Browning JD, Baker JA, Rogers T et al. Short-term weight loss and hepatic triglyceride reduction: evidence of a metabolic advantage with dietary carbohydrate restriction. Am J Clin Nutr. 2011 May; 93(5): 1048–1052.
Monday, 7 March 2016
The state of nutritional science, 2016
Someone sent me a link to this paper the other day, it's not something I would have looked at otherwise. However, next time you read a "totality of the evidence" snowjob supporting guidelines on saturated fat consumption, this paper will probably have been thrown on the pile, weighting it a little bit further.
Finucane OM, Lyons CL, Murphy AM et al (19 authors).
Monounsaturated fatty acid-enriched high-fat diets impede adipose NLRP3 inflammasome-mediated IL-1β secretion and insulin resistance despite obesity.
Diabetes. 2015 Jun;64(6):2116-28. doi: 10.2337/db14-1098. Epub 2015 Jan 27.
Abstract
Saturated fatty acid (SFA) high-fat diets (HFDs) enhance interleukin (IL)-1β-mediated adipose inflammation and insulin resistance. However, the mechanisms by which different fatty acids regulate IL-1β and the subsequent effects on adipose tissue biology and insulin sensitivity in vivo remain elusive. We hypothesized that the replacement of SFA for monounsaturated fatty acid (MUFA) in HFDs would reduce pro-IL-1β priming in adipose tissue and attenuate insulin resistance via MUFA-driven AMPK activation. MUFA-HFD-fed mice displayed improved insulin sensitivity coincident with reduced pro-IL-1β priming, attenuated adipose IL-1β secretion, and sustained adipose AMPK activation compared with SFA-HFD-fed mice. Furthermore, MUFA-HFD-fed mice displayed hyperplastic adipose tissue, with enhanced adipogenic potential of the stromal vascular fraction and improved insulin sensitivity. In vitro, we demonstrated that the MUFA oleic acid can impede ATP-induced IL-1β secretion from lipopolysaccharide- and SFA-primed cells in an AMPK-dependent manner. Conversely, in a regression study, switching from SFA- to MUFA-HFD failed to reverse insulin resistance but improved fasting plasma insulin levels. In humans, high-SFA consumers, but not high-MUFA consumers, displayed reduced insulin sensitivity with elevated pycard-1 and caspase-1 expression in adipose tissue. These novel findings suggest that dietary MUFA can attenuate IL-1β-mediated insulin resistance and adipose dysfunction despite obesity via the preservation of AMPK activity.
This was a complicated and expensive piece of work, with no obvious COIs:
The work presented in this article has been supported by Science Foundation Ireland http://dx.doi.org/10.13039/501100001602 (grant SFI PI/11/1119). The CORDIOPREV and LIPGENE study subjects and investigators were funded by European Commission FP6 (grant FOOD-CT-2003-505944).
So has it been designed in such a way that it can tell us anything reliably about the effects of fats in human diets?
- The mice are C57BL/6 so get fat on diets that may not fatten humans,[1] and the diet is 45% fat, 35% CHO which is half sugar, plus casein. Trisun oil vs Palm oil used means PUFA was well-controlled. However, it also means we're looking at a particular type of SFA - palmitate and stearate - versus the generic MUFA oleic acid that's prolific in every fat, even the palm oil here.The mice fed more MUFA gained less weight than the mice fed SFA. Is this plausible? It's consistent with Delaney et al (2000).[2] However, this same evidence would predict that, had the SFA source been coconut oil, then the SFA mice would have gained less weight than the MUFA mice.
- SFA/MUFA intake of first human cohort (CORDIOPREV) is determined by plasma levels, which evry ful kno are controlled by CHO,[3] and this is clearly shown in supplementary table 3 where plasma SFA correlates with triglycerides but not HDL. Therefore, CORDIOPREV shows effect of CHO intake on IR as well as or instead of effect of SFA intake. It is well-known that sugar and SFA consumption are associated in epidemiology, and that is the likely explanation for what we see in supplementary table 3.

- Second human study (LIPGENE) compares effect of MUFA intervention in high-SFA people living in Scandanavia with same intervention in high-MUFA people living in Italy. Again, SFA/MUFA was determined by plasma levels. Even apart from this, I think there could be a few confounders within this arrangement that were not discussed in the paper. Results are shown in supplementary figure 6 here.
Interestingly, there is no discussion whatsoever of limitations vs strengths of the research in the paper.
Does this mean the results are wrong? Many studies show that MUFA is associated with better insulin sensitivity than SFA in high-carb diets, but the difference seems to disappear at higher fat intakes.
[Edit 1/4/16: the science for this is much less convincing than I was given to believe. Most studies show no difference. The KWANU study only showed a non-significant difference which disappeared at 47% fat. The effect is only seen in humans, when it is seen, with differences in fat type that are outside the normal range obtained in people freely choosing fatty foods, in other words at the same extra-physiological concentrations of the various fatty acids that are produced in the mouse studies.]
The mouse study "demonstrated that enrichment of obesigenic HFDs with MUFA can improve insulin sensitivity, reduce adipose IL-1β–mediated inflammation, and promote adipose hyperplasia compared with diets enriched with SFA". Promoting adipose hyperplasia might not be something everyone wants. A possible explanation is that more fat storage in subcutaneous adipose tissue (and somewhat higher rate of oxidation) from MUFA in the obesigenic diet results in less visceral and ectopic fat - the former (VAT) is more inflammatory than subcutaneous fat, the latter (ectopic fat in liver and pancreas) results in insulin resistance and type 2 diabetes.[4] But because of the mouse model used, this doesn't answer the question, what is an obesigenic diet in humans? The C57BL/6 mouse can't tell us what happens in humans if MUFA replaces carbohydrate instead of replacing SFA, but we have human studies showing that, e.g.[5]
"As compared with the high-carbohydrate diet, the high-monounsaturated-fat diet resulted in lower mean plasma glucose levels and reduced insulin requirements, lower levels of plasma triglycerides and very-low-density lipoprotein cholesterol (lower by 25 and 35 percent, respectively; P less than 0.01), and higher levels of high-density lipoprotein (HDL) cholesterol (higher by 13 percent; P less than 0.005)."
[1] Borghjid S, Feinman R. Response of C57Bl/6 mice to a carbohydrate-free diet
Nutrition & Metabolism 2012;9:69
[2] DeLany, JP, Windhauser, MW, Champagne, CM, Bray, GA. Differential oxidation of individual dietary fatty acids in humans. Am J Clin Nutr October 2000;72(4): 905-911
[3] Volk BM, Kunces LJ, Freidenreich DJ et al. Effects of step-wise increases in dietary carbohydrate on circulating saturated fatty acids and palmitoleic acid in adults with metabolic syndrome. PLoS One. 2014 Nov 21;9(11):e113605. doi: 10.1371/journal.pone.0113605. eCollection 2014.
[4] Sattar N, Gill JMR. Type 2 diabetes as a disease of ectopic fat? BMC Medicine 2014; 12: 123
[5] Garg A, Bonanome A, Grundy SM, Zhang ZJ, Unger RH. Comparison of a high-carbohydrate diet with a high-monounsaturated-fat diet in patients with non-insulin-dependent diabetes mellitus. N Engl J Med. 1988 Sep 29;319(13):829-34.
Finucane OM, Lyons CL, Murphy AM et al (19 authors).
Monounsaturated fatty acid-enriched high-fat diets impede adipose NLRP3 inflammasome-mediated IL-1β secretion and insulin resistance despite obesity.
Diabetes. 2015 Jun;64(6):2116-28. doi: 10.2337/db14-1098. Epub 2015 Jan 27.
Abstract
Saturated fatty acid (SFA) high-fat diets (HFDs) enhance interleukin (IL)-1β-mediated adipose inflammation and insulin resistance. However, the mechanisms by which different fatty acids regulate IL-1β and the subsequent effects on adipose tissue biology and insulin sensitivity in vivo remain elusive. We hypothesized that the replacement of SFA for monounsaturated fatty acid (MUFA) in HFDs would reduce pro-IL-1β priming in adipose tissue and attenuate insulin resistance via MUFA-driven AMPK activation. MUFA-HFD-fed mice displayed improved insulin sensitivity coincident with reduced pro-IL-1β priming, attenuated adipose IL-1β secretion, and sustained adipose AMPK activation compared with SFA-HFD-fed mice. Furthermore, MUFA-HFD-fed mice displayed hyperplastic adipose tissue, with enhanced adipogenic potential of the stromal vascular fraction and improved insulin sensitivity. In vitro, we demonstrated that the MUFA oleic acid can impede ATP-induced IL-1β secretion from lipopolysaccharide- and SFA-primed cells in an AMPK-dependent manner. Conversely, in a regression study, switching from SFA- to MUFA-HFD failed to reverse insulin resistance but improved fasting plasma insulin levels. In humans, high-SFA consumers, but not high-MUFA consumers, displayed reduced insulin sensitivity with elevated pycard-1 and caspase-1 expression in adipose tissue. These novel findings suggest that dietary MUFA can attenuate IL-1β-mediated insulin resistance and adipose dysfunction despite obesity via the preservation of AMPK activity.
This was a complicated and expensive piece of work, with no obvious COIs:
The work presented in this article has been supported by Science Foundation Ireland http://dx.doi.org/10.13039/501100001602 (grant SFI PI/11/1119). The CORDIOPREV and LIPGENE study subjects and investigators were funded by European Commission FP6 (grant FOOD-CT-2003-505944).
So has it been designed in such a way that it can tell us anything reliably about the effects of fats in human diets?
- The mice are C57BL/6 so get fat on diets that may not fatten humans,[1] and the diet is 45% fat, 35% CHO which is half sugar, plus casein. Trisun oil vs Palm oil used means PUFA was well-controlled. However, it also means we're looking at a particular type of SFA - palmitate and stearate - versus the generic MUFA oleic acid that's prolific in every fat, even the palm oil here.The mice fed more MUFA gained less weight than the mice fed SFA. Is this plausible? It's consistent with Delaney et al (2000).[2] However, this same evidence would predict that, had the SFA source been coconut oil, then the SFA mice would have gained less weight than the MUFA mice.

- Second human study (LIPGENE) compares effect of MUFA intervention in high-SFA people living in Scandanavia with same intervention in high-MUFA people living in Italy. Again, SFA/MUFA was determined by plasma levels. Even apart from this, I think there could be a few confounders within this arrangement that were not discussed in the paper. Results are shown in supplementary figure 6 here.
Interestingly, there is no discussion whatsoever of limitations vs strengths of the research in the paper.
Does this mean the results are wrong? Many studies show that MUFA is associated with better insulin sensitivity than SFA in high-carb diets, but the difference seems to disappear at higher fat intakes.
[Edit 1/4/16: the science for this is much less convincing than I was given to believe. Most studies show no difference. The KWANU study only showed a non-significant difference which disappeared at 47% fat. The effect is only seen in humans, when it is seen, with differences in fat type that are outside the normal range obtained in people freely choosing fatty foods, in other words at the same extra-physiological concentrations of the various fatty acids that are produced in the mouse studies.]
The mouse study "demonstrated that enrichment of obesigenic HFDs with MUFA can improve insulin sensitivity, reduce adipose IL-1β–mediated inflammation, and promote adipose hyperplasia compared with diets enriched with SFA". Promoting adipose hyperplasia might not be something everyone wants. A possible explanation is that more fat storage in subcutaneous adipose tissue (and somewhat higher rate of oxidation) from MUFA in the obesigenic diet results in less visceral and ectopic fat - the former (VAT) is more inflammatory than subcutaneous fat, the latter (ectopic fat in liver and pancreas) results in insulin resistance and type 2 diabetes.[4] But because of the mouse model used, this doesn't answer the question, what is an obesigenic diet in humans? The C57BL/6 mouse can't tell us what happens in humans if MUFA replaces carbohydrate instead of replacing SFA, but we have human studies showing that, e.g.[5]
"As compared with the high-carbohydrate diet, the high-monounsaturated-fat diet resulted in lower mean plasma glucose levels and reduced insulin requirements, lower levels of plasma triglycerides and very-low-density lipoprotein cholesterol (lower by 25 and 35 percent, respectively; P less than 0.01), and higher levels of high-density lipoprotein (HDL) cholesterol (higher by 13 percent; P less than 0.005)."
[1] Borghjid S, Feinman R. Response of C57Bl/6 mice to a carbohydrate-free diet
Nutrition & Metabolism 2012;9:69
[2] DeLany, JP, Windhauser, MW, Champagne, CM, Bray, GA. Differential oxidation of individual dietary fatty acids in humans. Am J Clin Nutr October 2000;72(4): 905-911
[3] Volk BM, Kunces LJ, Freidenreich DJ et al. Effects of step-wise increases in dietary carbohydrate on circulating saturated fatty acids and palmitoleic acid in adults with metabolic syndrome. PLoS One. 2014 Nov 21;9(11):e113605. doi: 10.1371/journal.pone.0113605. eCollection 2014.
[4] Sattar N, Gill JMR. Type 2 diabetes as a disease of ectopic fat? BMC Medicine 2014; 12: 123
[5] Garg A, Bonanome A, Grundy SM, Zhang ZJ, Unger RH. Comparison of a high-carbohydrate diet with a high-monounsaturated-fat diet in patients with non-insulin-dependent diabetes mellitus. N Engl J Med. 1988 Sep 29;319(13):829-34.
Wednesday, 24 February 2016
Good News from Melbourne University's NZO Mouse LCHF "Paleo" Study
The egregious behaviour of Prof Sol Andrikopoulos in his press release to global media has obscured the finding that was the primary purpose of the "Three Mouseketeers" study.The study was supposed to answer a question that is actually of great importance in diabetes research:
- the blood sugars of people with diabetes usually improve and can become completely normal on the LCHF diet - this is actually something that has been known for a century or more.
- if you challenge someone who is doing well on the LCHF diet with an oral glucose tolerance test (a sudden large carbohydrate load) their response is often poor.
- is this a physiological adaptation to the diet, or does it indicate a risk of beta cell deterioration (as is usually seen long-term when high carb diets are fed to people with type 2 diabetes)?
The problem with the study, with regard to this question, is:
1) that the NZO mice chosen had a genetic defect, one that has never occurred in humans, which makes them fat-intolerant.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1693963/
2) the NZO LCHFD mice gained weight significantly, whereas most overweight humans lose weight on the LCHFD and it is very rare to gain a significant amount of fat mass.
Weight is an important determinant of insulin resistance and glycemic control.

4) the diets were in no way designed to test the Paleo premise, which is that specific Neolithic and refined and processed modern foods have deleterious effects on the human organism, including with regard to weight and glycemic control.
Thus the study was inconclusive on its own terms, and could shed little light on the effect of a LCHFD in humans, and press releases equating its results to the probable effect of a LCHF or a Paleo diet (2 different things) were wholly unjustified.
These remarks and the wide publicity they received amount to unjustified interference in diabetes treatment methods that are working well.
If we take the results at face value, and assume they do apply to humans, it's an interesting exercise.
Most humans with type-2 diabetes told to eat high-starch diets will suffer beta-cell damage in a few years time and require ongoing increases in dose and number of anti-hyperglycaemic medication. These mice gained weight and became insulin resistant on the LCHFD yet didn't suffer beta-cell damage (at least according to the tests chosen by the Three Mouseketeers).
If pre-diabetic mice that become fat and IR on a LCHF diet don't suffer beta-cell damage, maybe humans that lower fat and insulin won't either. Unlike most type 2 diabetic humans on high-carb diets. I don't see how one could be any worse off, anyway, and at least you'd always be eating the way that exposed you to least excess blood glucose and lowest risk of medication side effects.
As the authors say, "Indeed, there is mounting evidence that initial hypersecretion of insulin in prediabetes contributes to β-cell stress and failure."
If mice were men.
More here
http://profgrant.com/2016/02/20/the-most-famous-new-zealand-mice-in-the-world/
Wednesday, 27 January 2016
Saturated fat epidemiology - EPIC Netherlands and Malmö Diet and Cancer Study
The latest large epidemiologicial study on saturated fat and heart disease (IHD events) arrived yesterday. 35,597 people, followed for 12 years, suffered 1807 IHD events. It's called
"The association between dietary saturated fatty acids and ischemic heart disease depends on the type and source of fatty acid in the European Prospective Investigation into Cancer and Nutrition-Netherlands cohort." (full text here)
The finding? A small reduction in IHD events (heart attacks, angina, and such) in those eating most SFA from dairy foods and solid fats, no change with higher intakes from meat. The interesting thing is that they did one of those "substitute 5% of energy from saturated fat" analyses and, according to this data set, if you substitute saturated fat with PUFA, MUFA, lean protein (except vegetable protein) or high- or medium- GI carbhydrate (but not low-GI carbs) this predicts more IHD.
Why? Well the authors go on about a bit of trans fats in the oils and spreads. To me this makes little difference - the only reason people eat that crap is because they're trying to avoid (or can't afford) foods with more saturated fat. Also, those with higher SFA intakes weren't exactly avoiding foods likely to contain trans fat, just eating less of them, and the exposure wasn't huge by US standards (there's no CSPI in the Netherlands).
It seems more likely that these results (if they have any validity - this was only a small effect, in FFQ epdemiology, and it only crosses the centreline after adjustment) show the influence of food quality.
People eating most SFA ate more SFA from cheese, butter, and solid fats, and less SFA from snacks, soft and liquid fats, and "other" sources. Ergo they ate fewer erzatz foods and more real foods.
Anyway this is just FFQ epidemiology and it'll go into the next meta-analysis of saturated fat and heart disease and that correlation will become even closer to null than it already is.
But what would happen if you used a more reliable and time-consuming method, like a 7-day food diary, on a population of similar size and dietary habits?
Saturated Fat – the risks and benefits in a higher-fat
population.
The most common criticism, certainly the most serious we get is, that saying that butter and cream and full-fat dairy aren’t necessarily harmful in a low carb diet will cause people to eat more of these foods and increase the risk of heart disease. Actually we think that, in theory at least, a high intake of saturated fat of the type found in butter, palm oil, and red meat could well be harmful in the context of a diet high in refined carbs, like the Standard American Diet (SAD), or the diet a lot of Kiwis end up with when they eat cheap convenience food.
The most common criticism, certainly the most serious we get is, that saying that butter and cream and full-fat dairy aren’t necessarily harmful in a low carb diet will cause people to eat more of these foods and increase the risk of heart disease. Actually we think that, in theory at least, a high intake of saturated fat of the type found in butter, palm oil, and red meat could well be harmful in the context of a diet high in refined carbs, like the Standard American Diet (SAD), or the diet a lot of Kiwis end up with when they eat cheap convenience food.
In the past
epidemiological studies that have tried to answer the question about saturated
fat and heart disease have produced inconsistent results, with the aggregate (meta-analysis)
showing no correlation between saturated fat and cardiovascular disease or
total mortality.
One reason for this inconsistency between individual studies has been a failure to control for other variables, including trans fats. Another is the methods used to collect information – most studies in the past used the food frequency questionnaire (FFQ) which required subjects to guess how often they ate certain foods, which was then checked in an interview. Other more reliable methods that have been developed are the 4-day food diary, where the subject writes down the food they eat in real time.
Recently two new papers from the Malmö Diet and Cancer study caught our attention. This study has followed 26,930 people for 14 years, and dietary intake was assessed using a 7-day food diary and a 1 hour interview, as well as the FFQ. The study was also able to identify and exclude people who had a history of changing their diet, and intake of industrial trans fat was very low in the whole population. In the first study, both dairy fat consumption (including butter and cream) and intake of the shorter-chain saturated fats (4:0 – 14:0) found in dairy (and also in coconut, but that wasn’t a common food in Malmö) were associated with a significantly reduced incidence of type 2 diabetes over 14 years of follow up (about a 17% reduction overall).[1] In the second study, compliance with recommendations to reduce saturated fat intake to 14% of energy or less was associated with a 15% increase in diabetes in men, and a slightly smaller increase in women. Because of this effect of saturated fat reduction, the totality of “healthy eating” advice such as we see in NZ – eat less butter, more fish, more fruit and veges, more whole grains, and so on - had no effect on the incidence of diabetes.[2]
We were intrigued by this information, and we wondered what the Malmö Diet and Cancer study had to say about cardiovascular disease, given that it concerns a population eating more fat, and more dairy fat, than NZ, and given that the data collection methods used seemed to have been so much more reliable than those used in the past.
First of all a word about Sweden. The range of fat intake in Malmö when divided into quartiles, goes from about 30% of energy (the NZ recommendation) for the lowest quartile to 48% for the highest (which is practically low carb). The foods highest in specific fats in Sweden are – saturated fat: dairy and meat, monounsaturated fat: vegetable oil and meat, polyunsaturated fat: vegetable oils and spreads. The polyunsaturated fat quartiles range from 4% - 8% of energy, similar to NZ. Cooking fats are butter, vegetable oil, and a mysterious “cooking (or liquid) margarine”. This is not a trans fat source but sales have been declining recently, while sales of butter and oil have increased. A large proportion of the vegetable oils and spreads used in Sweden are canola-based. Sweden today has a lower rate of overall vascular mortality than New Zealand, and a similar rate of heart disease mortality.
When we look at fat and the main causes of mortality in Malmö, we find no correlation at all between saturated fat and any cause of death. Even the statistically insignificant correlations for CVD are in favour of saturated fat. When we look at total fat, there is an interesting variation. Men in the highest quartile for fat, at 47.7% of energy, have a 35% lower risk of dying of cardiovascular disease than men getting 31.7% of energy from fat.[3] There’s no effect of fat on cardiovascular disease in women, but higher fat consumption is associated with a 46% increased cancer mortality. This doesn’t correlate to saturated fat, or polyunsaturated fat, but to monounsaturated fat. Women in Malmö eat less fat from meat than men (who have no correlation between fat and cancer), so much of the monounsaturated fat may be coming from vegetable oils and liquid margarine. As Swedish polyunsaturated fat intakes are 4-8%, only a little above the natural range, the liquid margarine (made by Unilever) and cooking oils used will tend to be high-oleic lines.
Things get very interesting when the combined effect of saturated fat and fibre on cardiovascular mortality is considered. Saturated fat, which is mostly from dairy fat in Malmö, may have a protective effect against cardiovascular disease in people who eat the most fibre.[4] The main sources of fibre are vegetables and fruit, with a smaller amount from wholegrains. Men who eat high fibre, low saturated fat, or low fibre, high saturated fat, or high fibre, high saturated fat all have the same rate of ischemic CVD; but men in the lowest quintiles for both fibre and saturated fat combined have an 82% increased risk of iCVD, and there was also a significantly elevated risk in adjacent quintiles.
One reason for this inconsistency between individual studies has been a failure to control for other variables, including trans fats. Another is the methods used to collect information – most studies in the past used the food frequency questionnaire (FFQ) which required subjects to guess how often they ate certain foods, which was then checked in an interview. Other more reliable methods that have been developed are the 4-day food diary, where the subject writes down the food they eat in real time.
Recently two new papers from the Malmö Diet and Cancer study caught our attention. This study has followed 26,930 people for 14 years, and dietary intake was assessed using a 7-day food diary and a 1 hour interview, as well as the FFQ. The study was also able to identify and exclude people who had a history of changing their diet, and intake of industrial trans fat was very low in the whole population. In the first study, both dairy fat consumption (including butter and cream) and intake of the shorter-chain saturated fats (4:0 – 14:0) found in dairy (and also in coconut, but that wasn’t a common food in Malmö) were associated with a significantly reduced incidence of type 2 diabetes over 14 years of follow up (about a 17% reduction overall).[1] In the second study, compliance with recommendations to reduce saturated fat intake to 14% of energy or less was associated with a 15% increase in diabetes in men, and a slightly smaller increase in women. Because of this effect of saturated fat reduction, the totality of “healthy eating” advice such as we see in NZ – eat less butter, more fish, more fruit and veges, more whole grains, and so on - had no effect on the incidence of diabetes.[2]
We were intrigued by this information, and we wondered what the Malmö Diet and Cancer study had to say about cardiovascular disease, given that it concerns a population eating more fat, and more dairy fat, than NZ, and given that the data collection methods used seemed to have been so much more reliable than those used in the past.
First of all a word about Sweden. The range of fat intake in Malmö when divided into quartiles, goes from about 30% of energy (the NZ recommendation) for the lowest quartile to 48% for the highest (which is practically low carb). The foods highest in specific fats in Sweden are – saturated fat: dairy and meat, monounsaturated fat: vegetable oil and meat, polyunsaturated fat: vegetable oils and spreads. The polyunsaturated fat quartiles range from 4% - 8% of energy, similar to NZ. Cooking fats are butter, vegetable oil, and a mysterious “cooking (or liquid) margarine”. This is not a trans fat source but sales have been declining recently, while sales of butter and oil have increased. A large proportion of the vegetable oils and spreads used in Sweden are canola-based. Sweden today has a lower rate of overall vascular mortality than New Zealand, and a similar rate of heart disease mortality.
When we look at fat and the main causes of mortality in Malmö, we find no correlation at all between saturated fat and any cause of death. Even the statistically insignificant correlations for CVD are in favour of saturated fat. When we look at total fat, there is an interesting variation. Men in the highest quartile for fat, at 47.7% of energy, have a 35% lower risk of dying of cardiovascular disease than men getting 31.7% of energy from fat.[3] There’s no effect of fat on cardiovascular disease in women, but higher fat consumption is associated with a 46% increased cancer mortality. This doesn’t correlate to saturated fat, or polyunsaturated fat, but to monounsaturated fat. Women in Malmö eat less fat from meat than men (who have no correlation between fat and cancer), so much of the monounsaturated fat may be coming from vegetable oils and liquid margarine. As Swedish polyunsaturated fat intakes are 4-8%, only a little above the natural range, the liquid margarine (made by Unilever) and cooking oils used will tend to be high-oleic lines.
Things get very interesting when the combined effect of saturated fat and fibre on cardiovascular mortality is considered. Saturated fat, which is mostly from dairy fat in Malmö, may have a protective effect against cardiovascular disease in people who eat the most fibre.[4] The main sources of fibre are vegetables and fruit, with a smaller amount from wholegrains. Men who eat high fibre, low saturated fat, or low fibre, high saturated fat, or high fibre, high saturated fat all have the same rate of ischemic CVD; but men in the lowest quintiles for both fibre and saturated fat combined have an 82% increased risk of iCVD, and there was also a significantly elevated risk in adjacent quintiles.
1
Fibre, SFA and iCVD in men
In the case of women, things are a little different, and the figures vary a lot (maybe because women have less iCVD than men, so statistical effects are underpowered). Women who combined high saturated fat intake (4th quintile) with the highest fibre intake (5th quintile) had the only significant association, a 64% reduced risk of iCVD (the rate was the same for the 5th quintile of both saturated fat and fibre, but was non-significant).
(Note that the lowest quintile, at 13% saturated fat, was compliant with the 14% saturated fat or less recommendation that was associated with the 15% increase in type 2 diabetes; saturated fat intake in the 5th quintile was 22% of energy).
The authors concluded that “This study of a well-defined population, where SFA intake was high overall, provides little support for independent effects of specific macronutrients in relation to risk of ischemic CVD”, but that gender-specific interactions between nutrients may exist.
The gender difference is exaggerated (or highlighted if you prefer) by the fact that the high-fibre, low-SFA group was chosen as the reference point (1.0) for both men and women because it was anticipated – wrongly, as it turns out - that this was where the lowest risk would fall.

2Fibre,
SFA and iCVD women
As well as fibre, the Malmö Diet and Cancer study controlled for smoking, educational status, BMI, blood pressure, drug use (statins or blood pressure drugs), alcohol use, and activity.
Of these, educational status had a high independent correlation with carotid artery stenosis, a feature of atherosclerosis, in women. Women with lower levels of education or in manual jobs had about double the rate of carotid stenosis of those with a full secondary and tertiary education or clerical job (education is mandatory between the ages of 7 and 16 in Sweden), but the association was much weaker for men.[5]
Epidemiological studies will always be imperfect, and correlation definitely isn’t causation, but this study is as good as it gets, and the absence of correlation, which becomes stronger as time goes on and more studies come in (as shown by the latest meta-analysis) is not something we would expect to see if saturated fat plays a causal role in disease.[6] That would be contrary to the whole premise of epidemiology.
So – saturated fat isn’t associated with cardiovascular disease or mortality in a large population where intake is high, but varies a lot, and where dairy is the main source of saturated fat. Of course, we may be accused of cherry picking, there are a couple of other large modern studies that have used similar methods and that may be just as reliable that we haven’t looked at yet. But this criticism misses the point.
If saturated fat doesn’t kill people or cause heart disease in one place, or in another place, then why should we expect it to be lethal at our place?
In Malmö, people who liked cream on their berries, full-fat yoghurt on their fruit salad, who fried their leeks and cabbage in butter, and roasted carrots, beetroot, and brussels sprouts with their meat, and put butter and cheese on their rye bread, were apparently doing okay. And that’s what we should expect – we should expect people who’re eating well to be healthier than people who are eating poorly.
That – to understand how to eat well - used to be the basic purpose of nutritional science. And, when it was, the population had a much clearer idea of how to go about it. People knew how to cook because they were allowed and encouraged to cook the same foods their parents and grandparents cooked. Thanks to journalists with a historical interest and science training, like Gary Taubes (in Good Calories, Bad Calories and How We Get Fat) and Nina Teicholz (in The Big Fat Surprise) we now know how nutrition lost its way.
The question is, what will it take to get it back on the right path?
Of course, in promoting a low carb diet, we’re potentially exposed to the same criticisms as those who promoted the low-saturated fat diet.
However, there are important differences.
Limiting fat and saturated fat was supposed to reduce cardiovascular disease risk over a period of many years – it wasn’t supposed to make you feel better or reverse any health problems in the short term. You were supposed to limit saturated fat forever to get the benefit, and you needed to use some refined and additionally processed foods to do it, like oils, spreads, and low fat meat and milk products, not to mention cereal products.
Whereas the low carb diet has been shown to reverse some existing disease symptoms fairly rapidly, for example in the case of diabetes, and it often makes people feel better. And if a person, especially an insulin-sensitive, healthy person, tries a low carb diet for a while and then decides that some carbohydrate foods are in fact good for them after all, they may well be right. They’ll know more about the effect of carbohydrate foods on their body and will probably make better decisions about those foods from a nutritional point of view. Because, we’re not promoting a dietary change that increases your dependence on refined and processed foods. Whether you eat high or low carb, we don’t think that eating foods with a high HI (human interference) factor is a good idea. LCHF is a good way of reducing the HI factor in your diet, because the highest HI foods tend to be the sweet and starchy ones.
[1] Ericson, U, Hellstrand, S, Brunkwall, L, Schulz, C-A, Sonestedt, E, Wallström, P, et al. Food sources of fat may clarify the inconsistent role of dietary fat intake for incidence of type 2 diabetes. AJCN 2015;114.103010v1
http://ajcn.nutrition.org/content/early/2015/04/01/ajcn.114.103010
[2] Sonestedt, E, et al. A high diet quality based on
dietary recommendations does not reduce the incidence of type 2 diabetes in the
Malmo Diet and Cancer cohort. EADS2015 ePoster #322 http://www.easdvirtualmeeting.org/resources/a-high-diet-quality-based-on-dietary-recommendations-does-not-reduce-the-incidence-of-type-2-diabetes-in-the-malmo-diet-and-cancer-cohort--3
[3] Leosdottir, M, Nilsson, PM, Nilsson, J-Ã…, MÃ¥nsson, H, Berglund, G. Dietary fat intake and early mortality patterns – data from The Malmö Diet and Cancer Study. Journal of Internal Medicine
Volume 258, Issue 2, pages 153–165, August 2005.
http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2796.2005.01520.x/full
[3] Leosdottir, M, Nilsson, PM, Nilsson, J-Ã…, MÃ¥nsson, H, Berglund, G. Dietary fat intake and early mortality patterns – data from The Malmö Diet and Cancer Study. Journal of Internal Medicine
Volume 258, Issue 2, pages 153–165, August 2005.
http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2796.2005.01520.x/full
[4] Wallström P, Sonestedt E, Hlebowicz J, Ericson U,
Drake I, Persson M, et al. (2012) Dietary Fiber and Saturated Fat Intake
Associations with Cardiovascular Disease Differ by Sex in the Malmö Diet and
Cancer Cohort: A Prospective Study. PLoS ONE 7(2): e31637. doi:10.1371/journal.pone.0031637
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0031637
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0031637
[5] Rosvall, M, Östergren, PO, Hedblad, B, Isacsson,
S-O, Janzon, L, Berglund, G. Occupational Status, Educational Level, and the
Prevalence of Carotid Atherosclerosis in a General Population Sample of
Middle-aged Swedish Men and Women: Results from the Malmö Diet and Cancer Study.
Am J Epidemiol 2000;152:334–46
[6] de Souza, RJ, Mente, A, Maroleanu, A, Cozma, AI,
Ha, V, Kishibe,T, et al. Intake of saturated and trans unsaturated fatty acids
and risk of all cause mortality, cardiovascular disease, and type 2 diabetes:
systematic review and meta-analysis of observational studies. BMJ
2015;351:h3978
http://www.bmj.com/content/351/bmj.h3978
http://www.bmj.com/content/351/bmj.h3978
Sunday, 27 December 2015
FGF21 - a liver hormone linking sugar cravings and cardiovascular disease.
On Christmas Eve the media carried reports that scientists had identified a hormone, produced by the liver, that switches off sugar cravings, and which might be the answer to sugar addiction.
"Research on mice and monkeys has shown that the hormone, FGF21, signals the brain to avoid seeking sweet foods.
Harnessing the effect, possibly by copying the hormone's action with a drug, could help patients who are obese or suffering from Type 2 diabetes, scientists believe."
We all know that a LCHF diet suppresses the appetite for sugar - one of my dietary epiphanies involved standing in a supermarket, among shelves of garishly packaged chocolates and sweets, and realising that though being exposed to that stuff had always obliged me to buy some piece of it to take away and eat before, now I hadn't a single impulse worth fighting to buy or eat any of it ever again.
So it was an obvious question (one I may not have asked had I read the full paper first, because in the actual experiment FGF21 is clearly being produced in response to eating carbohydrates) whether a ketogenic diet raises FGF21.
It does - FGF21 is part of the regulatory response to fasting or a ketogenic diet (in mice).
So, eat keto, and your liver produces FGF21, which stops you wanting sweet food. Simple.
But, like that book by Ben Goldacre, I think you'll find it's a bit more complicated than that.
Luckily though, the complexity of FGF21 seems to fit a pattern we've seen before with other hormones.
While I was looking for the link to ketosis, I noticed in my search results a paper linking elevated FGF21 to cardiovascular disease. How can something that stops you from eating sugar be linked to CVD? That was before I knew that FGF21 was produced in response to eating carbohydrates. By the time I read the CVD paper, I knew what I was looking for. Here it is;
The metabolic syndrome has also been associated with
increased serum FGF21 levels, whereas an increase in FGF21
serum levels has been suggested as a new biomarker for
nonalcoholic fatty liver disease or steatohepatitis (17, 54, 60,
62, 116, 118). A study on obese children confirmed that
increased serum FGF21 is correlated to BMI and free fatty
acids (90). When serum FGF21 levels were tested after an oral
load of fructose, it was interestingly shown that FGF21 values
acutely spike, presenting a similar curve as serum glucose and
insulin after a glucose load. This finding shows that FGF21
presents a typical hormonal response possibly mediated by
carbohydrate-responsive element-binding protein that is activated
by fructose (18).
...it was shown that FGF21
levels are predictive of combined cardiovascular morbidity and
mortality (Fig. 3) (59). Increased baseline serum levels of this
molecule were found to be associated with a higher risk for
cardiovascular events in patients with type 2 diabetes in the
Fenofibrate Intervention and Event Lowering in Diabetes
(FIELD) study, and interestingly this association tended to be
stronger in the patient group that presented higher total cholesterol
levels (84). The authors speculate that the increased
basal levels of FGF21 in this group of patients may be an
indication of the potential role of FGF21 as a biomarker for the
early detection of cardiometabolic risk and furthermore that it
may reflect a compensatory response or the need of supraphysiological doses of FGF21 as a result of FGF21 resistance, a hypothesis proven in obese mouse models (21).
FGF21 resistance and hyperFGF21aemia. It's a familiar pattern. If FGF21 is produced in response to carbohydrate - maybe when hepatocytes reach glycogen saturation or ATP depletion or some other threshold - but your lifestyle or culture involves eating past the signal, maybe because of some dopamine effect of sugar you're sensitive to, or because you bought the 1.5 litre bottle of Coke because it was cheaper than the 300ml and it shouldn't go to waste, or because your mum or dietitian is telling you to finish your cake or keep eating the low fat food regularly, then maybe your liver eventually, because you stopped listening to it, makes so much FGF21 that the cells that should notice it become insensitive to it, so you eat more sweet carbs, make more FGF21, and get the same vicious cycle that we see with insulin.
(Or maybe there's some other cause for FGF21 resistance, a virus or environmental toxin or food colouring or genetic bad luck. It doesn't really matter unless you finish the cake.)
So what happens when a modern human goes on a keto diet or fasts? FGF21 may not rise at all - instead, sensitivity can be restored by its dropping, much like insulin.
Raymund Edwards astutely tweeted this study, which shows what happens to FGF21 when humans fast for 10 days. It's not the same as it was with the mice. In the mice on the ketogenic diet, it sometimes looked as if FGF21 played a role in ketogenesis - in the fasting humans, ketones rose first and FGF21 followed days later. This actually makes more sense, because ketones suppress appetite independently of FGF21, and are produced through basic biochemical economics - this shouldn't require some fancy new hormone, just let glucagon dominate over insulin and the Krebs cycle will do the rest.
In some of the humans, FGF21 was elevated at baseline and dropped fairly quickly, as can be seen in the spaghetti plot:

Whereas ketones rose more rapidly:

Anyway, this line of investigation, which I have only skimmed superficially here, gives us two possibilities; we have pathways which we can use to explain the loss of a sweet tooth when carbs are restricted (either FGF21 elevation, or the restoration of FGF21 sensitivity), and, we have an additional connection between sugar and cardiovascular disease.
What does it mean that FGF21 rises so much when fasting (and probably similarly on a keto diet), if elevated FGF21 is associated with CVD and other metabolic diseases?
If you read the CVD paper, FGF21 has a number of beneficial and antiatherogenic properties. It doesn't seem like bad stuff to have elevated, unless it got that way from a high intake of fructose.
"Research on mice and monkeys has shown that the hormone, FGF21, signals the brain to avoid seeking sweet foods.
We all know that a LCHF diet suppresses the appetite for sugar - one of my dietary epiphanies involved standing in a supermarket, among shelves of garishly packaged chocolates and sweets, and realising that though being exposed to that stuff had always obliged me to buy some piece of it to take away and eat before, now I hadn't a single impulse worth fighting to buy or eat any of it ever again.
So it was an obvious question (one I may not have asked had I read the full paper first, because in the actual experiment FGF21 is clearly being produced in response to eating carbohydrates) whether a ketogenic diet raises FGF21.
It does - FGF21 is part of the regulatory response to fasting or a ketogenic diet (in mice).
So, eat keto, and your liver produces FGF21, which stops you wanting sweet food. Simple.
But, like that book by Ben Goldacre, I think you'll find it's a bit more complicated than that.
Luckily though, the complexity of FGF21 seems to fit a pattern we've seen before with other hormones.
While I was looking for the link to ketosis, I noticed in my search results a paper linking elevated FGF21 to cardiovascular disease. How can something that stops you from eating sugar be linked to CVD? That was before I knew that FGF21 was produced in response to eating carbohydrates. By the time I read the CVD paper, I knew what I was looking for. Here it is;
The metabolic syndrome has also been associated with
increased serum FGF21 levels, whereas an increase in FGF21
serum levels has been suggested as a new biomarker for
nonalcoholic fatty liver disease or steatohepatitis (17, 54, 60,
62, 116, 118). A study on obese children confirmed that
increased serum FGF21 is correlated to BMI and free fatty
acids (90). When serum FGF21 levels were tested after an oral
load of fructose, it was interestingly shown that FGF21 values
acutely spike, presenting a similar curve as serum glucose and
insulin after a glucose load. This finding shows that FGF21
presents a typical hormonal response possibly mediated by
carbohydrate-responsive element-binding protein that is activated
by fructose (18).
...it was shown that FGF21
levels are predictive of combined cardiovascular morbidity and
mortality (Fig. 3) (59). Increased baseline serum levels of this
molecule were found to be associated with a higher risk for
cardiovascular events in patients with type 2 diabetes in the
Fenofibrate Intervention and Event Lowering in Diabetes
(FIELD) study, and interestingly this association tended to be
stronger in the patient group that presented higher total cholesterol
levels (84). The authors speculate that the increased
basal levels of FGF21 in this group of patients may be an
indication of the potential role of FGF21 as a biomarker for the
early detection of cardiometabolic risk and furthermore that it
may reflect a compensatory response or the need of supraphysiological doses of FGF21 as a result of FGF21 resistance, a hypothesis proven in obese mouse models (21).
FGF21 resistance and hyperFGF21aemia. It's a familiar pattern. If FGF21 is produced in response to carbohydrate - maybe when hepatocytes reach glycogen saturation or ATP depletion or some other threshold - but your lifestyle or culture involves eating past the signal, maybe because of some dopamine effect of sugar you're sensitive to, or because you bought the 1.5 litre bottle of Coke because it was cheaper than the 300ml and it shouldn't go to waste, or because your mum or dietitian is telling you to finish your cake or keep eating the low fat food regularly, then maybe your liver eventually, because you stopped listening to it, makes so much FGF21 that the cells that should notice it become insensitive to it, so you eat more sweet carbs, make more FGF21, and get the same vicious cycle that we see with insulin.
(Or maybe there's some other cause for FGF21 resistance, a virus or environmental toxin or food colouring or genetic bad luck. It doesn't really matter unless you finish the cake.)
So what happens when a modern human goes on a keto diet or fasts? FGF21 may not rise at all - instead, sensitivity can be restored by its dropping, much like insulin.
In some of the humans, FGF21 was elevated at baseline and dropped fairly quickly, as can be seen in the spaghetti plot:

Whereas ketones rose more rapidly:

Anyway, this line of investigation, which I have only skimmed superficially here, gives us two possibilities; we have pathways which we can use to explain the loss of a sweet tooth when carbs are restricted (either FGF21 elevation, or the restoration of FGF21 sensitivity), and, we have an additional connection between sugar and cardiovascular disease.
What does it mean that FGF21 rises so much when fasting (and probably similarly on a keto diet), if elevated FGF21 is associated with CVD and other metabolic diseases?
If you read the CVD paper, FGF21 has a number of beneficial and antiatherogenic properties. It doesn't seem like bad stuff to have elevated, unless it got that way from a high intake of fructose.
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