Rory Robertson and Protein Restricted Longevity

I looked at this paper when it did the rounds a fair while ago, saw that the only fat source used was soybean oil and decided that living on soybean oil, sucrose, maltodextrin and wheat starch was not a good idea and so I binned it as the junk it is:

The Ratio of Macronutrients, Not Caloric Intake, Dictates Cardiometabolic Health, Aging, and Longevity in Ad Libitum-Fed Mice

I missed the embedded problems which have since been brought to light by Rory Robertson, whose slightly over-the-top concerns are voiced here. I would perhaps disagree slightly with some of his opinions but, overall, he makes a rather good case. The first thing to note is that you have to go to the supplementary data to realise that a significant number of groups of mice were lost (and excluded) due to mortality problems. Table S1 describes all thirty of the diets which the study started out with. Five of these diets had to be discontinued because too many mice either died outright or (I suspect) were ordered to be euthanased on the authority of the supervising veterinary surgeon due to concerns about animal welfare. I'm assuming Oz has a Home Office much as the UK does which requires Named Veterinary Surgeons to be employed to supervise animal welfare in all laboratories.

We know this from the legend to supplementary table S1. Here is the last section of that legend (sorry that the small letter superscripting is lost, that's blogger for you):

"a Diets 2 low energy and 6 medium energy were discontinued within 23 weeks. b Diets 3 low energy, 3 medium energy and 6 low energy were discontinued within 10 weeks of treatment. These diets were discontinued due to weight loss (≥ 20%), rectal prolapse or failure to thrive".

Here is table S1 with the discontinued (and removed) groups outlined in red:

So, they started with 30 diets groups but five of them had such high early death rates that they were excluded from the study. This left 25 groups. Other than the legend to supplementary table S1 I am unable to find any reference to the loss of five diet groups anywhere in the main paper, which gives the distinct impression that 25 groups were all that were included from the start. So 17% of the mice died at under 23 weeks in to the study, many of those within less than 10 weeks, and you have to read the supplementary data to find out.

All of the high mortality groups were eating 5% of calories as protein.

Did you pick that up in the abstract? No, you didn't.

Is there any excuse for failing to discuss this crucial finding in the results and discussion sections of the paper? You can decide that. It's not exactly rocket science.

My feeling is that the authors could argue, if they were convinced that protein restriction was key to longevity (amino acids, cysteine, mTOR etc don'tchano), that studying early life mortality has no relevance to late life longevity. Why not leave early mortality to the paediatricians? That is a potentially arguable position and should, as it involved a huge chunk of the study mice, have been reported and been justified (if possible) in the results, discussion and especially in the abstract.

The other slight hiccup is this line from the main paper:

"Median lifespan was greatest for animals whose intakes were low in protein and high in carbohydrate... (Figure 2A)"

Figure 2A is not in a format which lends itself to simple interpretation and, obviously, excludes all of the mice which died or were euthanased at less than 23 weeks of age, all of which were in low protein groups. Anyway, you might want to see a simple table of median lifespan for each of the surviving groups. Like supplementary table S2. I've high-lighted the group which had the longest median lifespan in red:

Looks to me like the longest median lifespan group might have been eating 42% of it's calories as protein... Hmmmm. Worth repeating:

"Median lifespan was greatest for animals whose intakes were low in protein and high in carbohydrate... (Figure 2A)".

vs Table S2 giving 42% protein for longest median lifespan.

Hmmmmmmmmm...



Let's make this crystal clear: The data demonstrating the actual outcomes are, absolutely, present in the supplementary data of the paper. It is also absolutely crystal clear that the paper itself, excluding supplementary information, does not accurately represent the the actual findings in the study.

If you had to summarise the paper in human terms you could say that applying severe protein restriction to your kids while topping up their calories with sugar and soyabean oil would hopefully result in them being taken away from you and placed in to care before they died.

Please don't try this at home.

Peter

My thanks to Rory Robertson for his attempts to have this paper retracted and more accurately rewritten and to Grant Schofield for tweeting about his efforts.